THE 2019 EP ELECTION CAMPAIGN AS SEEN FROM TWITTER
The end of May 2019 saw European citizens go to the polls to choose a new European Parliament (EP). These elections marked a crucial moment for European democracy, as they took place in a context of significant challenges and upheavals, including the aftermath of the economic and financial crisis that had impacted the EU in the preceding decade, the refugee crisis that had tested the EU's capacity to manage migration and asylum, and the departure of the United Kingdom from the EU. These events had far-reaching implications for the political landscape of the EU, and the election campaign was shaped by the issues and concerns that they raised, as well as the parties and candidates who sought to address them. Against this backdrop, the 2019 EP election campaign was marked by a high degree of contentiousness and uncertainty, with many parties and candidates seeking to position themselves in response to the shifting political and economic landscapeMoreover, the 2019 EP election campaign was characterized by the significant role of social media in political communication, a phenomenon that had not been as prominent in previous campaigns. This transformation was due to the widespread dissemination of digital communication technologies, which had granted European citizens unprecedented access to information and new opportunities for political participation. As a result, political parties and politicians have greatly increased the number of available communication channels in order to disseminate information online and mobilize party members and supporters, making the use of social media a critical aspect of election campaign activity. Virtually every political party in Europe had a presence on social media, highlighting the importance of these platforms in modern-day political communication. The literature has recognized the significance of social media in shaping the outcome of election campaigns and has examined its role in creating new opportunities for political participation, promoting transparency and responsiveness between citizens and their representatives.An online election campaign offers a “window of opportunity” where behaviors and interactions between parties, candidates and citizens can be observed in condensed form. In the case of European Parliament elections, the view can embrace 28 different countries simultaneously in the same campaign context and in the same digital arena. Despite its many promises however the online dimension of EP campaigns is still relatively understudied. In this dissertation I present three distinct studies, each addressing a different dimension of the multifaceted and complex phenomenon of the 2019 EP online campaign.In the first chapter I examine the role of candidates and parties in online election campaigns, given the increasing importance of social media platforms. Specifically, the study investigates the prevalence of Twitter usage among candidates for the 2019 European Parliament election, as well as the factors that influence adoption and use of the platform. The study takes a comparative approach, analyzing data from all 28 member states of the European Union, and it aims to contribute to the ongoing “equalization vs. normalization” debate on the impact of social media on election campaigns. In particular, I investigate whether social media can be viewed as a force that equalizes the resource gaps between candidates and parties, or its use is being normalized into the EP campaign. To accomplish this, the study considers both individual-level and party-level factors that may influence Twitter adoption and use during the EP 2019 campaign.Chapter 2 investigates the dynamic nature of online communication between European Parliament (EP) candidates and their audiences during the 2019 EP campaign in the five largest Western European countries: France, Germany, Italy, Spain, and the United Kingdom. The study uses onl
THE 2019 EP ELECTION CAMPAIGN AS SEEN FROM TWITTER / G. Pagano ; supervisor: L. CURINI; direttore PhD: M. Jessoula. Dipartimento di Scienze Sociali e Politiche, 2022 Oct 25. 34. ciclo, Anno Accademico 2021.
INJECTABLE HYBRID HYDROGELS FOR LOCAL CHEMOTHERAPY OR TUMOR-SPECIFIC RESECTION
The aim of my thesis is to develop cutting-edge and innovative delivery system able to encapsulate active molecules against cancer. The purpose is to have a material that can target directly the tumoral environment, avoiding the toxicity to surrounded organs and enhancing the local pharmacological activity.Moreover, the use of a polymeric material, that can serve as a scaffold to deliver nanoparticles crosslinked between the network can increase the efficacy and bioavailability. For this reason, using both chemical and physical crosslinking strategies to make a hybrid material between polymer and nanoparticles could be investigated as a potential material used for minimally invasive surgery and for cancer treatment. The controlled drug release and the targeted delivery are essential parameters that need to be controlled in order to have active compound in a tumoral environment released at a certain dose over the time.
INJECTABLE HYBRID HYDROGELS FOR LOCAL CHEMOTHERAPY OR TUMOR-SPECIFIC RESECTION / A. Ajo ; tutor: L. De Cola ; phd coordinator: G. Vistoli. - Milano. Dipartimento di Scienze Farmaceutiche, 2024 May 20. 36. ciclo, Anno Accademico 2023.
EARLY CARNIAN AMMONOIDS FROM NEVADA: REVISED TAXONOMY, EVOLUTIONARY TRENDS AND USEFULNESS FOR THE DEFINITION OF THE LATE TRIASSIC TIME SCALE
The aim of this research is the study of the Lower Carnian (Upper Triassic) ammonoid faunas from Nevada, which have contributed to the definition of the North American Triassic chronostratigraphic scale in the 1960s. These faunas, however, have never been described in detail, both in terms of taxonomy and in terms of stratigraphic distribution in bed-by-bed sampled sections. This research is a contribution for the revision and improvement of the Late Triassic chronostratigraphic time scale. A revised zonation is proposed for the sedimentary succession within the middle member of the Augusta Mountain Formation at South Canyon (New Pass Range, Nevada), type locality of the Desatoyense Zone of the North American chronostratigraphic scale. New Lower Carnian ammonoid faunas from China Mountain (Tobin Range, Nevada) are also described. Silberling & Tozer (1968) introduced the Desatoyense Zone to replace the “Joannites zone” of Johnston (1941), with Trachyceras desatoyense Johnston, 1941, as the index species. The fauna described by Johnston (1941) was collected without accounting for the stratigraphic position of the specimens, and consequently, a range chart could not be provided for the 24 taxa attributed to this zone. An intensive sampling program in the type locality was begun in 2002 by Marco Balini and James Jenks and continued intermittently with other occasional collaborators over a period of several years. Bed by bed collection efforts from 103 fossiliferous beds in six stratigraphic sections have yielded ~3900 specimens, of which 3130 ammonoids, that allow the construction of range charts and recognition of the bioevents on which this revision is based. The systematic study has led to the recognition of 29 taxa, 10 of which are new. These taxa belong to 18 genera, two of which are new: Neoclypites, Sageceras, Perrinoceras, Badiotites, Procladiscites, Arcestes, Joannites, Silenticeras, Frankites, Daxatina, Trachyceras, Genus A, Genus B, Clionitites, Hannaoceras, Sympolycyclus, Lobites and Coroceras. Ammonoids of the Subfamily Trachyceratinae are the most abundant, and provide all the zonal indexes. Frankites sutherlandi and Daxatina occur in the lower part of the succession (sections A and B). The middle-upper part of the succession (section D2, E and lower part of section F) exhibits a different faunal composition and Trachyceras co-occurs with Genus A and Genus B (both Trachyceratinae), which are confined to two narrow stratigraphic intervals. The upper part of section F is characterized by another major faunal change, the onset of Trachyceras with highly indented suture lines that co-occur with Coroceras. With the new data it is possible to subdivide the former Desatoyense Zone into three parts, which are in stratigraphic order: the Daxatina sp. A range zone, the Trachyceras desatoyense interval zone and the Trachyceras sp. B range zone. The Daxatina sp. A zone is correlated with the Canadian Sutherlandi Zone and with the Tethyan Canadensis Subzone. The T. desatoyense and the T. sp. B zones are correlated with the Canadian Desatoyense Zone and with the Tethyan Aon Subzone. The Desatoyense Zone is reported also from China Mountain (Tobin Range, Nevada). This area was investigated in the early 1970s as part of the PhD work of K.M. Nichols (1972). Nichols collected ammonoids from two sites (localities #1251 and #1311, Nichols, 1972), but did not report bed-by-bed collection data, nor described the collection. Nichols & Silberling (1977), referred to the locality and assigned the ammonoid faunas to the Desatoyense Zone. During the 2018 field campaign we visited the area, and noted the location of the fossiliferous interval in the uppermost part of the Smelser Pass Member of the Augusta Mountain Formation, a few meters below the erosional base of the Cane Spring Formation. Two new fossiliferous sites were located and sampled bed-by-bed from exposed beds, and float spec
"EARLY CARNIAN AMMONOIDS FROM NEVADA: REVISED TAXONOMY, EVOLUTIONARY TRENDS AND USEFULNESS FOR THE DEFINITION OF THE LATE TRIASSIC TIME SCALE / R. Marchesi ; tutor: M.Balini ; coordinator: F. Camara Artigas. DIPARTIMENTO DI SCIENZE DELLA TERRA ""ARDITO DESIO"", 2020 Feb 04. 32. ciclo, Anno Accademico 2019. [10.13130/marchesi-ruben_phd2020-02-04]."
MICROSCOPIC THEORY OF INFINITE NUCLEAR MATTER AND NON-EMPIRICAL ENERGY FUNCTIONALS
Nuclear density functional theory (DFT) and ab initio theory are complementary approaches to nuclearstructure. DFT can be applied to the whole nuclear chart, but relies on empirical energy density functionals(EDFs). As a consequence, predictions in regions where experimental data are lacking, e.g. in neutron-richnuclei close to the driplines, are subject to significant uncertainties. In contrast, ab initio theory allows to drawpredictions with controlled uncertainties, but there are difficulties in extending these methods to heavy nucleior e.g. far from magic numbers. The purpose of this thesis is to combine DFT and ab initio by grounding thenuclear EDFs into ab initio theoretical predictions, specifically on infinite nuclear matter calculations.The ab initio Quantum Monte Carlo (QMC) and Self-consistent Green’s functions (SCGF) methods are employedto study nuclear matter. An extension of SCGF based on the algebraic diagrammatic construction(ADC-SCGF) is developed to include Gorkov pairing correlations. The ab initio equations of state (EOS) areused to construct EDFs based on the local density approximation and gradient approximation, which are appliedto magic nuclei. Then, the static response of nuclear matter is tackled with both DFT and QMC, and astudy of the constraints that can be set on the EDF surface terms from ab initio is reported.This thesis outlines a new strategy to ground nuclear EDFs into ab initio and implements some of its pillars.A critical discussion of the difficulties encountered is given. Future developments include the study ofnuclear matter from a microscopic perspective, e.g. extending ADC-SCGF to tackle the dynamic response andsuperfluid matter.
MICROSCOPIC THEORY OF INFINITE NUCLEAR MATTER AND NON-EMPIRICAL ENERGY FUNCTIONALS / F. Marino ; supervisor: G. Colò ; cosupervisor: F. Pederiva : coordinatore: R. Vecchi. Dipartimento di Fisica Aldo Pontremoli, 2023. 36. ciclo, Anno Accademico 2023.
GM1 OLIGOSACCHARIDE MODULATION OF CALCIUM SIGNALLING IN NEURONAL FUNCTIONS
It has been already demonstrated that the oligosaccharide chain (OligoGM1) of the ganglioside GM1, β-Gal-(1-3)-β-GalNAc-(1-4)-[α-Neu5Ac-(2-3)]-β-Gal-(1-4)-β-Glc-(1-1)-Ceramide, promotes neurodifferentiation in the Neuro2a murine neuroblastoma cells, used as a model, by directly interacting with the NGF specific receptor TrkA, leading to the activation of ERK1/2 downstream pathway. In this context, my PhD work aimed to investigate which other biochemical pathways, in addition to TrkA-MAPK cascade activation, are prompted by OligoGM1, with an emphasis on Ca2+ modulating factors.A proteomic analysis (nLC-ESi-MS-MS) performed on Neuro2a cells treated with 50 µM OligoGM1 for 24 hours led to the identification and quantification of 324 proteins exclusively expressed by OligoGM1-treated cells. Interestingly, some of these proteins are involved in the regulation of Ca2+ homeostasis and in Ca2+-dependent differentiative pathways. In order to evaluate if OligoGM1 administration was able to modulate Ca2+ flow, we performed calcium-imaging experiments on Neuro2a cells using the Ca2+-sensitive Fluo-4 probe. Starting from 5 minutes upon OligoGM1 administration to undifferentiated Neuro2a, a significant increase in Ca2+ influx occurs. At the same time an increased activation of TrkA membrane receptor was observed and, importantly, the addition of a specific TrkA inhibitor abolished the OligoGM1 mediated increase of the cytosolic Ca2+, suggesting that the opening of the cell Ca2+ channels following OligoGM1 administration depends on the activation of TrkA receptor. To unveil which cellular pathway activated by OligoGM1 could lead to the increase of intracellular Ca2+, time-course immunoblotting analyses were performed. The data revealed that following TrkA activation, OligoGM1 induced the activation of phospholipase PLCγ1 which converts phosphatidylinositol 4,5-bisphosphate (PIP2) to diacyglycerol (DAG) and inositol 1,4,5-trisphosphate (IP3), the second messengers that propagate cellular signalling via Ca2+ mobilization. Moreover, we observed a hyperphosphorylation of the DAG substrate, protein kinase C (PKC), which is a priming event that enables its catalytic activation in response to lipid second messengers, and we found its enrichment in lipid rafts, events that consolidate its activation. When calcium-imaging experiments where performed in the presence of xestospongin C, a potent inhibitor of IP3 receptors on endoplasmic reticulum, a reduction of about 50% of Ca2+ influx was observed, suggesting that the Ca2+ flows moved by the OligoGM1 come not only from intracellular storages, but probably also from the extracellular environment. Accordingly, in the presence of both extracellular (EGTA) and intracellular (BAPTA-AM) Ca2+ chelators the neuritogenic effect induced by OligoGM1 was abolished. The work described in this thesis confirms that the effects of GM1 ganglioside on neuronal differentiation are mediated by its oligosaccharide portion. In particular, here I highlight that the oligosaccharide, initiating a signalling cascade on the cell surface, is responsible alone for the balancing of the intracellular Ca2+ levels that underlie neurite sprouting, which have historically been attributed to the whole GM1 ganglioside and its role as lipid inserted into the plasma membrane. In this way, these data give additional information on the molecular characterization of the mechanisms by which GM1 exerts its neuronal functions.
GM1 OLIGOSACCHARIDE MODULATION OF CALCIUM SIGNALLING IN NEURONAL FUNCTIONS / G. Lunghi ; tutor: A. Prinetti, S. Sonnino ; supervisore: E. Chiricozzi ; coordinatore: A. Prinetti. Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, 2020 Dec 09. 33. ciclo, Anno Accademico 2020. [10.13130/lunghi-giulia_phd2020-12-09].
FACILITATING CLINICAL DECISION-MAKING IN REHABILITATION: INNOVATIVE APPROACHES TO SYNTHESIZE AND TRANSLATE EVIDENCE INTO PRACTICE
Integrating scientific evidence into clinical rehabilitation practice is essential to ensuring effective, data-driven interventions. However, translating this evidence into everyday clinical decision-making presents significant challenges due to the complexity of available information and the need to personalize interventions for individual patients. This project aims to explore innovative approaches to facilitate clinical decision-making in rehabilitation, focusing on synthesizing and applying available evidence to practice.Building upon the fundamental principle that rehabilitation interventions do not directly target the disease itself but rather address impairments and activity/participation levels, as influenced by the underlying health condition, and acknowledging the lack of direct evidence on rehabilitation for post-COVID-19 condition (PCC), we developed a series of overviews of Cochrane Systematic Reviews (CSRs). These overviews employ a mapping synthesis approach, focusing on the rehabilitative management of key PCC symptoms that have been previously investigated in other health conditions. The mapping syntheses allowed the identification of high-quality evidence relevant to the rehabilitation of fatigue, post-exertional malaise, orthostatic intolerance, dyspnea, arthralgia, dysphagia, dysphonia, olfactory dysfunction, cognitive impairment, anxiety, and depression. The findings have contributed to generating valuable hypotheses for both clinical practice and future research in PCC rehabilitation. Furthermore, these insights have served as the foundation for developing treatment recommendations for PCC symptoms, as published in the World Health Organization Guidelines for clinical practice. Thereafter, we conducted a systematic review to synthesize the increasing direct evidence on rehabilitation in the context of COVID-19 and PCC, providing practical recommendations for clinicians. This review serves as a valuable resource for informing clinical decisions and promoting the adoption of evidence-based practices.Finally, we identified a series of clinical conditions of high rehabilitative interest using a Delphi method. Subsequently, after developing a new methodological framework for conducting overviews of CSRs with a mapping synthesis, we applied this methodology to one of these relevant conditions (knee replacement rehabilitation).In conclusion, evidence mapping and systematic reviews represent an innovative approach to facilitating clinical decision-making in rehabilitation. These tools support practitioners in synthesizing and applying available evidence, promoting personalized, evidence-based interventions to improve patient outcomes.
FACILITATING CLINICAL DECISION-MAKING IN REHABILITATION: INNOVATIVE APPROACHES TO SYNTHESIZE AND TRANSLATE EVIDENCE INTO PRACTICE / C. Cordani ; tutor: S. Negrini; coordinatore: M. Del Fabbro. - Milano. Dipartimento di Scienze Biomediche, Chirurgiche ed Odontoiatriche, 2025 Dec 19. 38. ciclo, Anno Accademico 2024/2025.
Creative commons - http://creativecommons.org/licenses/by-sa/4.0/
THE CHALLENGE OF DOMAIN SHIFT IN REAL-WORLD ROBOTIC VISION: TOWARD SCALABLE, UNSUPERVISED, AND CLOUD-BASED ADAPTATION
Mobile robots are an emergent technology more and more present in contexts such as homes, offices, and hospitals to assist humans in daily life activities. Given the complexity of human-centric environments, robotic vision, namely computer vision embedded in mobile robots, has become an essential capability to acquire a semantically-rich understanding of the environment for improving core robotics tasks (such as navigation and localization), but also to enable high-level activities such as manipulation or human-robot interaction. Given the recent advances of deep learning, the naive solution to implement robotic vision is to leverage publicly available deep neural networks that can be mounted in robotic platforms with limited effort. Despite being widely adopted, this approach suffers from important limitations caused by the so-called domain shift problem: being trained on simulated or generic datasets (source domain), deep neural networks dramatically fail to tackle the complexity of the real-world environments (target domain) in which the robots operate. This dissertation investigates the challenges of domain shift in robotic vision and proposes novel adaptation strategies to enable robust and scalable real-world deployments of mobile robots. The contributions are structured into three complementary parts. First, we analyze the limitations of the mainstream pipeline to implement robotic vision that combines pre-trained neural networks with manual fine-tuning. We propose photorealistic simulation-based pre-training using data compliant with the robot's perception modality, and we demonstrate that fine-tuning with limited high-quality manual annotations substantially increases the model's robustness in the specific operational environment of the robot. Second, we remove the need for human supervision, proposing two alternative approaches for unsupervised model's adaptation. Our methods exploit the spatial constraints between neural network's predictions and the 3D world to enhance the quality of the pseudo-labels, thus enabling self-supervised adaptation. Third, we address adaptation in cloud-based robotic perception, where intensive inference required by neural networks is offloaded to remote servers to deal with the limited hardware configuration of mobile robots. In this context, we design solutions to ensure the scalability of domain adaptation and enable privacy preservation, designing lightweight neural networks that can be run locally by the robot. We validate our approaches using both real-world datasets and extensive experiments with real robotic platforms, considering multiple perception tasks such as object detection, 3D pose estimation, and semantic segmentation.
THE CHALLENGE OF DOMAIN SHIFT IN REAL-WORLD ROBOTIC VISION: TOWARD SCALABLE, UNSUPERVISED, AND CLOUD-BASED ADAPTATION / M. Antonazzi ; supervisor: N. Basilico ; co-supervisor: M. Luperto ; coordinator: R. Sassi. Dipartimento di Informatica Giovanni Degli Antoni, 2025 Dec 09. 38. ciclo, Anno Accademico 2025/2026.
A NOVEL RIT2-LRRK2 AXIS MODULATES LYSOSOME FUNCTION: INSIGHT FOR PARKINSON'S DISEASE PATHOPHYSIOLOGY
Lysosome dysfunction is recognized as a critical factor in Parkinson’s disease (PD) pathogenesis. Leucine-rich repeat kinase 2 (LRRK2) has a kinase-dependent role in autophagy and lysosomal function, and its pharmacological inhibition is protective against neurodegeneration and α-synuclein (αSyn) pathology. However, the precise signaling pathways involving LRRK2 and how alterations lead to pathology have not been clarified yet. In this work, we unravel a novel functional interaction between LRRK2 and the small GTPase Rit2, previously reported to participate in MAPK-signaling, neurite outgrowth and dopamine transporter trafficking. Polymorphisms in the RIT2 locus are associated to increased risk of PD and are predicted to alter gene expression. We found that RIT2 mRNA levels are reduced in substantia nigra pars compacta (SNpc) dopaminergic neurons of idiopathic PD patients and in different in vitro and in vivo PD models. Rit2 overexpression in G2019S-LRRK2 neuroblastoma cells restores lysosomal defects and diminishes αSyn accumulation, phenocopying pharmacological LRRK2 kinase inhibition. Moreover, Rit2 selective overexpression in SNpc dopaminergic neurons in A53T-αSyn virally injected mice reduces neurodegeneration and αSyn pathology. Notably, we uncovered that Rit2 interacts with LRRK2, and its overexpression inhibits LRRK2 kinase activity both in vitro and in vivo. In addition, Rit2 is required for lysosomal function, since its gene silencing in cell lines and DA neurons leads to lysosomal defects. We showed promising preliminary data indicating that the mechanism of Rit2 action in autophagy might involve lysosomal calcium release, that represents a convergent point in LRRK2-mediated regulation of autophagy. We propose Rit2 and its interplay with LRRK2 as novel targets for future, disease-modifying therapeutic approaches in PD.
A NOVEL RIT2-LRRK2 AXIS MODULATES LYSOSOME FUNCTION: INSIGHT FOR PARKINSON'S DISEASE PATHOPHYSIOLOGY / S. Pizzi ; co-tutor: M. Volta ; scientific supervisor: G. Cappelletti. Dipartimento di Bioscienze, 2023 Mar 30. 35. ciclo, Anno Accademico 2022.
THE ROLE OF MELT/OLIVINE RATIO IN DISSOLUTION AND REACTIVE CRYSTALLIZATION: AN EXPERIMENTAL AND MICROSTRUCTURAL STUDY (ELECTRON BACK-SCATTERED DIFFRACTION - EBSD) AT 0.5 GPA.
The role of melt/olivine ratio in dissolution and reactive crystallization: an experimental and microstructural study (Electron Back-Scattered Diffraction - EBSD) at 0.5 GPa.ABSTRACTMicrostructural and geochemical studies have emphasized that melt-rock reactions play a paramount role in the origin of gabbroic rocks in the lower oceanic crust. It is widely accepted that olivine-rich troctolites might form through dunite infiltrated followed by reactive crystallization of interstitial melts. Furthermore, experiments on the origin of olivine-rich troctolites have suggested that melt/olivine ratio is a fundamental factor in basalt-dunite interactions (Borghini et al., 2018). However, the effect of melt/rock ratio on the chemistry and the development of microstructures induced by reactive dissolution and crystallization processes have not been well defined yet. This work aims to experimentally evaluate and quantify the effect of melt/olivine ratio on mineral chemistry, melt evolution and microstructural development of olivine-rich troctolites formed by reactive processes. Electron Back-Scattered Diffraction (EBSD) analysis gives the opportunity to parametrize microstructures as a function of different melt/olivine ratio. Ten experiments were performed in piston cylinder apparatus at 0.5 GPa using San Carlos olivine (Fo90) and a MORB-type glass (AH6) as starting materials, mixed together in different proportions (i.e., the Initial Melt Amount, IMA, was equal to 10, 25 or 50wt%). Out of ten experiments, seven were isothermal runs at 1200, 1250 and1300°C, lasted 24 hours, and three were step-cooled runs carried out by lowering the temperature, with a fix rate of 1°C/min, from an isothermal dwell (at 1300°C) down to 1100°C.Run products of isothermal experiments at 1200°C consist of olivine, plagioclase, clinopyroxene and traces of glass, whereas at 1300°C consist only of olivine and glass. Texturally we distinguish two types of olivine: i) large subhedral crystals (up to100 µm) with both straight and lobate rims and ii) smaller (5-20 µm) rounded grains. Olivine grain size increases with the IMA; on contrary, the amount of smaller rounded grains is inversely correlated. At 1300°C, high IMA enhances the melt-olivine reaction, promoting olivine dissolution and crystal growth that leads to an increase in the olivine grain boundaries tortuosity. At 1200°C, mineral chemistry of olivine suggests that XMg values are not correlated with different IMA. At 1300°C, XMg in olivine increases from 0.89 ± 0.01 to 0.91 ± 0.01 with the increase of the IMA. The NiO content in olivine is strongly temperature dependent. At 1300°C, the NiO content in olivine decreases with the IMA. Clinopyroxene presents XMg = 0.89 ± 0.00IMA10wt%., 0.89 ± 0.02IMA25wt% and 0.84 ± 0.02IMA50wt%. Plagioclase has XAn = 0.68 ± 0.03IMA10wt%, 0.61 ± 0.06IMA25wt%. and 0.66 ± 0.02IMA50wt%. Reacted glasses at 1200°C have higher SiO2 and lower XMg than glasses at 1300°C, thus suggesting a strong control of olivine and interstitial phases on the residual glass composition. EBSD analysis suggests that the interaction between olivine matrix and basalt at 50wt%IMA and 1300°C strongly affects the pre-existing dunite matrix recovering its plastic deformation (i.e., deformed olivines grains underwent an annealing re-crystallization), thus generating an “undeformed” olivine-rich matrix. The average intragranular deformation parameter defined by the Grain Orientation Spread (GOS), indicates that no differences occur between the two different olivine morphologies. The smaller and rounded olivine has GOS = ~ 0.49 and the larger one with straight and lobate rims has GOS value = ~ 0.41. Hence, in terms of deformation grade, the assessment of a pre-existing olivine grain (Olivine1) and a new crystallized olivine (Olivine2) is not straightforward. Step-cooled experiments result in dunite (10wt%IMA), olivine-rich troctolite (25wt%IMA) an
THE ROLE OF MELT/OLIVINE RATIO IN DISSOLUTION AND REACTIVE CRYSTALLIZATION: AN EXPERIMENTAL AND MICROSTRUCTURAL STUDY (ELECTRON BACK-SCATTERED DIFFRACTION - EBSD) AT 0.5 GPA / M. Grammatica ; tutor: P. Fumagalli; co-tutors: G. Borghini, G. Capitani ; coordinatore: F. Camara Artigas. Università degli Studi di Milano, 2022 Feb 17. 34. ciclo, Anno Accademico 2021.
PEPTIDOGLYCAN REMODELLING FACTORS: KEY PLAYERS IN CELL DIVISION AND HOMEOSTASIS MAINTENANCE OF ESCHERICHIA COLI
In Escherichia coli, the structural integrity and coordinated growth of the cell envelope are crucial for survival, especially under environmental stress conditions. The lipopolysaccharide-containing outer membrane (OM) acts as a selective barrier against toxic molecules and provides additional mechanical support that complements the role traditionally attributed to the cell wall. Coordination between the OM and peptidoglycan (PG) growth is essential for maintaining cell envelope stability, particularly during cell division, when septal PG must be cleaved to allow daughter cell separation. This process is mediated by PG amidases, AmiA, AmiB, and AmiC, that are activated by LytM-domain proteins both from inside the cell, with the periplasmic protein EnvC that is anchored to the divisome components FtsEX, and the outside of the cell thanks to the OM lipoproteins NlpD and ActS. In the first manuscript included in this thesis, it is shown the identification of the periplasmic deacetylase SddA, responsible for modifying the amidase cleavage products. The expression of the non-essential sddA gene is linked with that of EnvC, highlighting the involvement of SddA in cell separation. Consistently with this speculation, the overexpression of sddA leads to the formation of long chains of unseparated cells, indicating that amidase activity at the septum is inhibited.The amidase activator ActS also displays a central role in envelope stress conditions. It has been shown that when OM biogenesis is disrupted, DpaA, the enzyme that detaches the OM lipoprotein Lpp from PG, is critical for preventing cell lysis. The lysis phenotype of dpaA deleted cells with defective LPS export is suppressed by the deletion of actS, suggesting that under LPS stress DpaA is capable of modulating ActS activity. A second manuscript included in this thesis shows that a DpaA-mediated modulation occurs at the ActS protein level.Together, these findings reveal a complex regulatory mechanism linking OM and PG growth, septal PG cleavage, and cell envelope remodeling under stress. This coordination ensures structural stability and viability of E. coli by integrating responses to envelope stress with the mechanical demands of cell division.
PEPTIDOGLYCAN REMODELLING FACTORS: KEY PLAYERS IN CELL DIVISION AND HOMEOSTASIS MAINTENANCE OF ESCHERICHIA COLI / M. Zaccaria ; tutor: A. Polissi ; phd coordinator: S. Ricagno. Dipartimento di Scienze Farmacologiche e Biomolecolari, 2024 Jan 10. 37. ciclo, Anno Accademico 2023/2024.
SYNTHESIS OF N-HETEROCYCLES VIA PALLADIUM-CATALYZED REDUCTIVE CYCLIZATION REACTIONS OF NITROARENES AND NITROALKENES USING FORMIC ACID DERIVATIVES AS CO SURROGATES
The reductive cyclization reaction of nitroarenes to produce N-heterocycles has been ‎investigated for more than four decades using CO as a cheap reductant, but it is still a ‎fascinating area for research and advancements. On the other hand, using toxic CO gas is ‎accompanied by many disadvantages. Thus, the thesis focuses on the use of phenyl formate and ‎formic acid as cheap, affordable, efficient and safe in-situ CO surrogates for the reductive ‎cyclization of various nitroarenes. Notably, in most cases, the isolated yield of the desired ‎heterocycle was higher than those previously obtained for the same reaction when gaseous CO ‎had been employed, which indicates that the use of these surrogates should not necessarily be ‎considered a second choice when the use of CO gas is not possible. A cheap thick-walled glass ‎‎“pressure tube” can be used instead of less-available autoclaves.‎
SYNTHESIS OF N-HETEROCYCLES VIA PALLADIUM-CATALYZED REDUCTIVE CYCLIZATION REACTIONS OF NITROARENES AND NITROALKENES USING FORMIC ACID DERIVATIVES AS CO SURROGATES / M.a.f. Abdellatif ; tutor: F. A. C. Ragaini ; coordinator: D. Passarella. Dipartimento di Chimica, 2024 Jan 31. 35. ciclo, Anno Accademico 2023/2024.
DEVELOPMENT OF PHYSIOLOGICAL CULTURE SYSTEMS TO ACCESS UNTAPPED OVARIAN RESERVES FOR IMPROVING REPRODUCTIVE EFFICIENCY
Female fertility preservation via complete in vitro folliculogenesis is still chimerical.Recreating folliculogenesis to grow a dormant primordial follicle to the preovulatorystage in vitro remains the most coveted goal of fertility preservation programs. Theability to grow undifferentiated oocytes in vitro from primordial follicles would increasethe supply of fully grown oocytes destined for downstream applications in clinicalsettings and the livestock industry. To date, the production of live offspring through thein vitro culture of primordial follicles has only been achieved in mice. While thisdemonstrates the potential value of primordial follicles as a source of fully grownoocytes, it remains a proof of principle. In large mammals, such as humans andbovine, in vitro follicle culture systems to produce mature oocytes from primordialfollicles are still experimental due to follicle mortality after isolation from thesurrounding tissue. While understanding the cause of follicle death is necessary toinhibit the induction of programmed cell death, it is also essential to elucidate themechanisms responsible for steering primordial follicles toward the following stages ofdevelopment, i.e., primary and secondary follicles. Mechanistically resolving bothsides of the story – primordial follicle death and development - would then provideinsight into crucial factors responsible for deciding the fate of the follicles and fuelingtheir further development in culture. Compilation of the information from transcriptomestudies would then inform the appropriate optimizations of culture systems andimprove the growth of isolated primordial follicles to the subsequent primary andsecondary stages entirely in vitro. To probe our objectives, we first laid the groundworkby developing a mechanical isolation protocol and serum-free base in vitro system toculture isolated bovine primordial follicles. We then quantitatively and qualitativelyassessed the isolation efficiency of primordial, primary, and secondary folliclepopulations from a defined volume of the ovarian cortex, thereby providing aconservative strategy to improve assisted reproductive technologies dedicated tofemale fertility preservation. Following this, interdisciplinary methods were adopted toelucidate the web of interacting genes governing primordial follicle fate in vitro and invivo. The conducted transcriptome and network analyses indicate that ferroptosis andautophagy are the elected programmed cell death mechanisms active in isolatedprimordial follicles cultured in vitro.Furthermore, we delineate the essential genes and pathways orchestrating theconsecutive developmental transitions (primordial-to-primary and primary-tosecondaryfollicles) in vivo through RNA sequencing of isolated primordial, primary,and secondary follicles. To extend our findings, we investigated the conservation offunctionally enriched pathways across different species. In conclusion, the studiesconducted during this doctoral program, provide a high-yield protocol that accessesuntapped ovarian reserves, thereby increasing the source of gametes for fertilitypreservation. We further define a base system for the in vitro culture of isolated bovineprimordial follicles and shortlist candidate molecular targets that can be modulated tooptimize physiological culture systems, thereby informing the overarching aim of thisresearch thesis.
DEVELOPMENT OF PHYSIOLOGICAL CULTURE SYSTEMS TO ACCESS UNTAPPED OVARIAN RESERVES FOR IMPROVING REPRODUCTIVE EFFICIENCY / P. Dey ; tutor: A. M. Luciano co-tutor: F. Franciosicoordinator: F. Ceciliani. Dipartimento di Medicina Veterinaria e Scienze Animali, 2024 May 29. 36. ciclo, Anno Accademico 2023.
BLOCKCHAIN & DATA JUSTICE. THE POLITICAL CULTURE OF TECHNOLOGY
Blockchain is a distributed ledger technology arising from the world of Bitcoin and cryptocurrencies, which has been advocated as a disruptive and revolutionary innovation. Because of the peculiarity of its architectural technicalities, in fact, blockchain technology has seen a growth of its social applications in recent years and has gained the attention of a large variety of actors, interested in its potentiality for entering the social domain and enabling the creation of decentralized, horizontal and peer-to-peer networks.This thesis aims at studying the visions of the world surrounding the implementation of blockchain technology, by analyzing narratives and discourses emerging among those who create and use it. Based on a critical understanding of algorithms and technology as non-neutral and as subject to bias and pitfalls, this work focuses on the sociological and political significance of imagining a technology for the social and public good, posing a particular attention to developers and digital entrepreneurs’ understanding of sociality.Drawing on a one-year multi-sited fieldwork in Milan, London, Tallinn and the online sphere, this work combines a theoretical analysis with ethnographical insights that arise from participant observation and a number of interviews with individuals from the blockchain scene. By relying on a qualitative study that aims at researching the values and aspirations that are encoded in technology, I argue that blockchain social imaginary is embedded in neoliberal, technocratic visions of the world and that blockchainers’ understanding of society and social relations becomes tokenized and subject to mathematical simplifications.My argument develops in three phases: firstly, I show how blockchain works as a floating signifier and thus could be interpreted as a populist buzzword; secondly, I argue that blockchain is surrounded by regimes of truth regarding its disruptive potential that overlooks social dynamics and logics; thirdly, I show that blockchainers’ understanding of social good is based on metrics and competition, thereby reinforcing a number of neoliberal myths associated with the Californian Ideology.By showing the importance of integrating more sociological perspectives to the study of digital technology’s potential, this work exits the financial and informatics domain, merging previous studies on blockchain with a human-rights based approach that ground its roots on social justice theory.
BLOCKCHAIN & DATA JUSTICE. THE POLITICAL CULTURE OF TECHNOLOGY / S. Semenzin ; tutor: P. A. Rebughini, A. Gandini ; coordinatori: M. Barisione, G. Semi. Università degli Studi di Milano, 2021 May 05. 33. ciclo, Anno Accademico 2020. [10.13130/semenzin-silvia_phd2021-05-05].
IL CONTROLLO DELLE CONCENTRAZIONI NELL¿UNIONE EUROPEA A OLTRE TRENT¿ANNI DALLA SUA INTRODUZIONE: LA RICERCA DI UN NUOVO MODELLO TRA SPINTE TECNOLOGICHE E NUOVE STRATEGIE POLITICHE
This thesis presents the results of a study on the shortcomings and reform perspectives of EU merger control rules, currently laid down by Regulation (EC) 139/2004. In the institutional and regulatory framework of the EU, merger control plays an atypical role, perhaps minor vis-à-vis the other tools of EU competition law, as proved by the absence of a wide-ranging literature (at least national). Merger control has, however, become the subject of an increasingly heated debate, involving scholars, institutional actors, and private parties, which concerns its effectiveness in the global, digitalized, and dynamic economy of this era. This debate, enthused by some socio-political events of great resonance, has been partially internalized by the EU institutions in the context of a wide inquiry, which is still at an early stage. Many of the needs for reform that have been voiced reflect, as a matter of fact, some tensions which have characterized the foundational and development stages of EU merger control. These tensions will be described in Chapter I, which will illustrate the sensitivity of EU merger control to the different approaches and objectives that emerged in the different phases of the European integration. Chapter II is, then, devoted to a critical examination of the role played by public interests (both national and supranational) within the system of EU merger control. Finally, Chapter III aims to provide an answer to the many questions posed by the digitization and the emergence of a new model of regulation of power relations among undertakings in the platform economy, which prevent a timely and appropriate intervention by the European Commission vis-à-vis concentrations that are potentially harmful to competition.
IL CONTROLLO DELLE CONCENTRAZIONI NELL¿UNIONE EUROPEA A OLTRE TRENT¿ANNI DALLA SUA INTRODUZIONE: LA RICERCA DI UN NUOVO MODELLO TRA SPINTE TECNOLOGICHE E NUOVE STRATEGIE POLITICHE / G. Dalla Valentina ; tutor: M. Condinanzi ; coordinatrice: F. Biondi. Università degli Studi di Milano, 2022 Jun 06. 34. ciclo, Anno Accademico 2021.
DTI-ALPS REFLECTS WHITE MATTER DIFFUSION ANISOTROPY AND MICROSTRUCTURAL HETEROGENEITY RATHER THAN GLYMPHATIC FLOW: INSIGHTS FROM MULTI-MODAL MRI
The glymphatic system plays a critical role in the clearance of metabolic waste and pathological proteins such as amyloid-β and tau from the brain, and its dysfunction has been increasingly implicated in the pathogenesis of neurodegenerative diseases, making its in vivo characterization a major research priority. The diffusion tensor imaging–analysis along the perivascular space (DTI-ALPS) has been widely adopted as a putative non- invasive MRI biomarker of glymphatic flow. However, the biological specificity of the DTI- ALPS index remains unclear, and recent evidence has raised concerns that it may primarily reflect white-matter (WM) microstructure rather than perivascular fluid transport. In this study, we sought to clarify the determinants of DTI-ALPS and to assess whether this metric primarily reflects WM microstructural damage rather than glymphatic activity.A total of 100 participants were recruited from individuals referred to the Neurodegenerative Diseases Unit of the Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico in Milan (Italy) for suspected dementia. All the participants underwent the same 3T MRI protocol comprising high-resolution T1-weighted and FLAIR imaging, double-shell diffusion-weighted imaging (b=1000 and 2000 s/mm2), and multi-echo gradient-echo sequences for quantitative susceptibility mapping. Within standard centrum semiovale regions of interest, we computed DTI-ALPS together with fractional anisotropy (FA), mean diffusivity (MD), mode of anisotropy (MA), and neurite orientation and density imaging (NODDI) metrics, including the orientation dispersion index (ODI), neurite density index (NDI), and isotropic volume fraction (FISO). We also computed the T1/FLAIR signal ratio and the diamagnetic component of susceptibility (DCS) derived from DECOMPOSE within the same ROIs, as complimentary structural metrics of WM damage.Lower DTI-ALPS values were associated with higher FA and MD (|r|=0.40–0.60), with stronger effects observed at b=2000 than at b=1000. The DTI-ALPS index correlated strongly and inversely with MA (r=–0.84 at b=1000; r=–0.86 at b=2000) and positively with ODI (r=0.73), and both metrics remained strong independent predictors after correction for age and sex. Moderate associations were observed with structural markers—direct with the T1/FLAIR ratio (r 0.33) and inverse with DCS standard deviation (r -0.50) —consistent with altered myelin integrity and increased microstructural heterogeneity. Exploratory factor analysis demonstrated that DTI-ALPS did not form a separate latent component but clustered with MA and ODI, indicating shared variance related to axonal geometry rather than to perivascular flow.Collectively, these findings demonstrate that DTI-ALPS predominantly reflects WM architecture—specifically, loss of crossing fibers, increased diffusion coherence, and reduced orientation dispersion—rather than the dynamics of glymphatic transport. Although DTI-ALPS may still capture secondary effects of glymphatic impairment arising from WM injury, the present results suggest that it cannot be considered a direct biomarker of glymphatic function. Its use as a surrogate measure of glymphatic flow should therefore be approached with caution and re-evaluated through direct validation against intrathecal contrast-enhanced MRI or other physiological reference methods.
DTI-ALPS REFLECTS WHITE MATTER DIFFUSION ANISOTROPY AND MICROSTRUCTURAL HETEROGENEITY RATHER THAN GLYMPHATIC FLOW: INSIGHTS FROM MULTI-MODAL MRI / L. Sacchi ; tutor: D. Galimberti ; tutor clinico: A. Arighi ; direttore del corso: M. Del Fabbro. Dipartimento di Scienze Biomediche, Chirurgiche ed Odontoiatriche, 2025 Dec 19. 38. ciclo, Anno Accademico 2024/2025.
STRUCTURAL ANALYSIS, RATIONAL DESIGN, AND OPTIMIZATION OF NOVEL MICROTUBULE TARGETING AGENTS.
Microtubule targeting agents represent a significant class of drugs, with currently more than20 approved formulations utilized in anti-inflammatory, anti-infective, and predominantlyanti-cancer therapies. These agents interact with a,b-tubulin heterodimers withinmicrotubules, impacting their dynamic behavior which is crucial in cellular processes likesignaling, transport, or cell division. Despite the extensive range of molecules studied andtrialed, central challenges in their therapeutic application remain, such as high toxicity,particularly neuropathies, low efficacy at tolerable doses as well as drug resistance. Efforts toaddress these issues include the characterization of new binding sites and molecules whichdemonstrate exceptional activity in resistant cell lines. This thesis contributes to this ongoingresearch, focusing on the structural characterization of maytansine-site ligands and the designof ligands for the Todalam site.
STRUCTURAL ANALYSIS, RATIONAL DESIGN, AND OPTIMIZATION OF NOVEL MICROTUBULE TARGETING AGENTS / A. Abel ; tutor: S. Pieraccini (UNIMI), D. Passarella (UNIMI), M. Steinmetz (University of Basel), A. Prota (Paul Scherrer Institute), J. Abrahams (University of Basel) ; coordinator: D. Roberto (UNIMI). Dipartimento di Chimica, 2024 Mar 14. 35. ciclo, Anno Accademico 2022.
In this PhD thesis we investigate several aspects of parton distribution functions (PDFs) andjets as applicable to the physics underpinning the Large Hadron Collider (LHC) as well as futurecolliders. We first discuss jet observables at the LHC, focusing on the single-jet inclusive crosssection. We introduce possible alternative definitions, which weigh the individual contributionscoming from each jet in the event and are thus unitary by construction. We also clarify theorigin of some problematic aspects of the standard definition. Secondly, within the hadronicPDF fitting framework of the NNPDF collaboration, we investigate the inclusion of single-jetinclusive and dijet measurements into a global PDF fit, using QCD next-to-next-to-leading orderpredictions for jet processes. We field-test which observables lead to better perturbative stability,better PDF compatibility with other data, better fit quality, and more stringent constraints onthe PDFs. Thirdly, we focus on an analytical understanding of machine learning techniques usedfor quark versus gluon discrimination, a hot topic in jet substructure studies. We construct anew version of the widely used N -subjettiness variable, which features a simpler theoreticalbehaviour than the original one, while maintaining, if not exceeding, the discriminating power.We input these new observables to the simplest possible neural network, with only one neuron,and we study analytically the network behaviour at leading logarithmic accuracy. We alsocompare our analytic findings to a more realistic neural network trained with Monte Carlopseudo-data. Fourthly, we compute the unpolarised electron, positron, and photon PDFs at next-to-leading logarithmic accuracy in QED, which are crucial for high-precision predictions neededfor future e + e − colliders. We present both numerical and analytical results. The analyticalpredictions, defined by means of a specific additive formula, provide a large-z analytical solutionthat includes all orders in the QED coupling constant α, with a small- and intermediate-zsolution that includes terms up to O(α 3 ). The content of this thesis is based on arXiv:1906.11850,arXiv:1911.12040, arXiv:2005.11327, and arXiv:2007.04319.
SUBSTRUCTURE AT COLLIDERS / G. Stagnitto ; tutor: F. Stefano, M. Cacciari (Université de Paris), G. Soyez (IPhT Saclay); coordinatore Scuola di Dottorato: M. Paris. Dipartimento di Fisica Aldo Pontremoli, 2020 Sep 14. 33. ciclo, Anno Accademico 2020. [10.13130/stagnitto-giovanni_phd2020].
TUBEROUS SCLEROSIS COMPLEX: IDENTIFICATION OF THE GENETIC CAUSE IN PATIENTS WITH NO MUTATION DETECTED, AND ANALYSIS OF MOSAICISM
Tuberous sclerosis complex (TSC) is an autosomal dominant multisystem disorder characterized by development of hamartomas, intellectual disability, seizures and autism. TSC is caused by inactivating mutations in either the TSC1 or the TSC2 genes. A pathogenic variant is not identified in up to 10% of the patients with a clinical diagnosis of TSC despite full molecular assessment. These individuals are referred to as NMI (No Mutation Identified), and it is not clear if they should be monitored and/or treated in similar fashion to those with known etiology of TSC.To identify the genetic cause of TSC in these patients, we selected ten individuals with a definite clinical diagnosis of TSC and NMI and performed a pilot study. Three different technologies were used and their results were compared: chromosomal microarray, trio whole genome sequencing, and targeted deep-coverage next generation sequencing of TSC1 and TSC2. We identified mosaic variants in TSC1/TSC2 in six patients. No variants in other genes were detected in the remaining individuals.Based on the results of the pilot study and on recent literature, we then performed targeted deep-coverage TSC1/TSC2 sequencing on 200 affected individuals using peripheral blood DNA and saliva or other tissue samples where available. We identified 24 patients with mosaic pathogenic variants in TSC1 (n=2) or TSC2 (n=22), defining a rate of mosaicism of 12%. Mosaic variant allele frequency (VAF) ranged from 2% to 32% in blood and from 2% to 35% in saliva. Most affected individuals had low-level mosaicism (VAF ≤10%). We performed an extensive analysis of the phenotype, and show that individuals with a mosaic variant in TSC1/TSC2 often display normal cognitive functioning, although other TSC-associated neuropsychiatric disorders (TAND) are seen in 62% of the patients. Cortical tubers are invariably present and seizures are diagnosed in 54% of the cohort, but infantile spasms are rare. The number of cutaneous manifestations in these patients is often insufficient to meet diagnostic criteria, except for facial angiofibromas. We observed a high frequency of pulmonary and renal manifestations in our mosaic cohort, which are as severe as those seen in the general TSC population. None of the patients who have reproduced transmitted the variant to their offspring.In conclusion, our study shows that genome sequencing fails to identify rare variants in new genes related to TSC, and a third gene is therefore unlikely to exist. We demonstrated that at least one out of 10 patients with a clinical diagnosis of TSC carries a mosaic pathogenic variant in TSC1 or TSC2. We also showed that individuals with mosaic variants have a distinctive phenotypic severity, with important implications for surveillance.
TUBEROUS SCLEROSIS COMPLEX: IDENTIFICATION OF THE GENETIC CAUSE IN PATIENTS WITH NO MUTATION DETECTED, AND ANALYSIS OF MOSAICISM / A. Peron ; tutor: M.P. Canevini ; coordinatori del dottorato: E. Berti, M. Del Fabbro. Dipartimento di Scienze della Salute, 2021 Dec 14. 34. ciclo, Anno Accademico 2021.
LA MOBILITÀ TRANSNAZIONALE DEI LAVORATORI. CONTRIBUTO ALLO STUDIO DEI PROFILI GIUSLAVORISTICI E DI SICUREZZA SOCIALE
The object of this final dissertation is to provide an analysis of the evolving and debated phenomenon of transnational posting of workers. At first, posting of workers is analysed in a private international law dimension, since the rules established by directive 96/71/CE are mandatory rules as described in art. 9 of Rome I regulation (chapter I). Posting of workers is then investigated in its relevant EU regulatory framework, starting from the case by case approach of the European Court of Justice in the 80s. The jurisprudence of the early stage and its implication are dealt with in chapter II. According to directive 96/71/EC, a “posted worker” is an employee who is sent by his employer to carry out a service in another EU Member State on a temporary basis, in the context of a contract of services, an intra-group posting or a hiring out through a temporary agency. The scope of the directive is to provide, in addition to the application of the sending State law, i.e. the habitual place of work, a set of mandatory rules of the host State so that to grant the posted worker an “hard core of minimum protection”. On the other hand, the directive aims at providing a level-playing field in the market of the provision of services. The above described rules have proved not to be effective to prevent social dumping and therefore the European legislator promoted the recent adoption of the Directive 2018/957/EU amending the Posting of Workers Directive 96/71/EC. The new approach undertaken by the EU legislator with the establishment of the principle of ‘same pay for the same work at the same workplace’ for posted and local workers is evaluated deeply in chapter III. The exam of the legal framework of the social security provisions in the EU is followed by a critical definition of the consequences of those provisions on the effectiveness of the social protection of posted workers (chapter IV). As to the conclusive remarks, a selection of possible legal instruments which would help to build a better protection of the posted workers are analysed, taking in consideration the suggestions of the European trade Unions and the academic community.
LA MOBILITÀ TRANSNAZIONALE DEI LAVORATORI. CONTRIBUTO ALLO STUDIO DEI PROFILI GIUSLAVORISTICI E DI SICUREZZA SOCIALE / G. Morra ; tutor: A. Tursi ; coordinatore del dottorato: G. Ludovico. Dipartimento di Diritto Privato e Storia del Diritto, 2021 Feb 22. 33. ciclo, Anno Accademico 2020.
THE PROBLEM OF WORK IN AXEL HONNETH'S CRITICAL THEORY
This thesis shows that although Axel Honneth's recognition based analysis of market-mediated social labour is resilient, it still requires further refinement and improvement. The objective of this thesis is to contribute to this end. Firstly, the thesis provides a qualified internal critique of Honneth's account, incorporating three elements: a) a systematic discussion of his arguments covering his early interventions on the topic up to and including his more recent writings; b) defence of Honneth's normative approach showing that the recognition theory provides robust rejoinders to functionalist objections, and to objections challenging the adequacy of his normative reconstruction method; c) a critique showing that Honneth's account has limited critical import in the context of contemporary 'ethicized' capitalism. The critique is based on the assessment that: i) recognition and autonomy, i.e. the norms underpinning Honneth's model representing preconditions of social freedom, have been integrated into the recent forms of work organisation while being re-signified in the process; ii) divorced from the moral grammar of social freedom norms acquire an ambivalent character and lose their critical potential. Secondly, the thesis shows that a recognition-based critique of 'ethicized' capitalism can be recovered should changes be made to Honneth's standard recognition model. The thesis argues that, in addition to Honneth's claim that social actors need primary forms of recognition to realise their social freedom, moral self-realisation should be acknowledged as an additional condition of social freedom and a possibility of social labour. Further, it shows that the relationship between work, moral responsibility, and recognition is relevant for a critique of work because novel forms of work organisation face workers today with increased demands for autonomy and responsibility while denying them organisational preconditions for assuming moral responsibility for their work, i.e. co-determination rights. Because 'ethicized' capitalism is underpinned by the promise of moral self-realisation, which it systematically fails to fulfil, moral responsibility acquires critical potential.
THE PROBLEM OF WORK IN AXEL HONNETH'S CRITICAL THEORY / A.f. Deaconu ; tutor: M. Geuna ; coordinator: A. Pinotti. Dipartimento di Filosofia Piero Martinetti, 2024 Jan 09. 35. ciclo, Anno Accademico 2022.
USE OF ON-FARM DATA AND MILK LEUCOCYTE COMPONENTS FOR MONITORING THE HEALTH OF DAIRY COWS
The aim of this PhD project was monitoring cow health and welfare, using on-farm data and leucocytes components in milk. For this reason, the investigation focused on the two main diseases of dairy cows: mastitis and lameness. Critical aspects are linked with these diseases and some of them were considered and discussed in four different experiments. The first experiment focused on the variability of leucocytes in milk, that affects its use as a detection tool for mastitis. For this reason, the factors that influence neutrophils, lymphocytes and macrophages of milk were analyzed. A total of 848 milk samples from 179 dairy cows bred in 6 farms of Northern Italy were collected and analyzed, with Vetscan DC-Q milk Analyzer (Advanced Animal Diagnostic) and DeLaval Cell Counter (DeLaval), to detect Total Leucocyte Count (TLC), Somatic Cell Count (SCC) and the concentration of neutrophils, macrophages and lymphocyte. The results of the Generalized Mixed Model highlighted that the three fractions of leucocytes were significantly influenced by parity, stage of lactation, and farm. The highest values of neutrophils (>60% of TLC) were found at the beginning and at the end of lactation, the two most critical moments for mastitis onset. In accordance, high level of neutrophils (>63% of TLC) was described as one of the risk factors of high SCC (>100,000 cells/ml), highlighting the possibility to apply neutrophils as indicators of udder issues, after accounting for parity, stage of lactation and management conditions. The second experiment focused on the reduction of antimicrobials usage to treat and prevent the insurgence of intramammary infections, investigating one of the main critical periods for antimicrobial overuse: dry-off. This experiment proposed a Selective Dry Cow Therapy (SDCT) protocol based on SCC (thresholds 100,000 cells/ml for primiparous cows and 200,000 cells/ml for multiparous ones) and Differential Somatic Cell Count (threshold of 69.3%) to identify the cows to treat and not treat at dry-off, with the aim to investigate the effects on udder health (SCC value). 33% of 243 dairy cows were selected to not be treated at dry-off. The analysis of milk sample at the beginning of lactation highlighted similar udder conditions, SCC (P=0.5) and TLC (P=0.7) were not different between cows treated with or without antimicrobials. On the other hand, the abandonment of antimicrobial therapy at dry-off, together with poor hygiene conditions and long dry periods (> 55 days), was identified as a risk factor of high SCC (>100,000 cells/ml) at the beginning of the subsequent lactation. The proposed SDCT protocol permits to select cows to treat and not treat with antimicrobials at dry-off, reducing efficiently antimicrobials usage without compromising udder health, even if adequate hygiene conditions and proper management of dry period is requested for the successful of the procedure.The last two experiments were focused on lameness and on-farm data were explored as early indicators of lameness onset. The aim was to produce a starting point for future studies investigating the possibilities of lameness detection without farmers needed to dedicate more work or incur in additional expenses. In the first experiment, milking (e.g. milk production, milking flows) and behavioral (e.g. lying time, number of steps, lying bouts) data were collected for one year, using only on-farm sensors (Afimilk) in one farm of Northern Italy. From the starting dataset, 3459 daily observations of 67 lame cows were extracted. Generalized Additive Mixed Models were used to model the trends of milking and behavioral parameters in the 30 days before and after the detection of lameness, and derivatives were used to identify the changing points of the function. The results showed a change in most of the parameters before the lameness detection, with different magnitude for the three parity groups of cows considered. The reduction of average milk
USE OF ON-FARM DATA AND MILK LEUCOCYTE COMPONENTS FOR MONITORING THE HEALTH OF DAIRY COWS / S. Mondini ; tutor: M. Zucali ; co-tutor: G. Gislon ; coordinatore: S. R. Pilu. Dipartimento di Scienze Agrarie e Ambientali - Produzione, Territorio, Agroenergia, 2025 Dec 16. 38. ciclo, Anno Accademico 2024/2025.
NOVEL MONO-, BI- AND TRI-SPECIFIC IMMUNOMODULATORY ANTIBODIES FOR CANCER THERAPY AND TOOLS FOR OTHER THERAPEUTIC AND DIAGNOSTIC APPLICATIONS
Cancer represents one of the leading causes of death worldwide, and the development of novel immunotherapeutic strategies, such as that of immune checkpoint inhibitors (ICIs), has generated long-lasting responses in multiple malignancies. Even though safer than chemotherapies, some adverse events, including rare cases of cardiotoxicity, have also been reported for ICIs, especially when used in combination.In this context, the aim of my research project was to identify combinations of novel human immunomodulatory monoclonal antibodies targeting different ICs, with improved efficacy and safety profiles. Some novel ICIs were previously isolated in our laboratory and characterized for their biological properties. Here, they were tested in combinations to identify those with stronger anti-tumor effects and reduced cardiotoxic side effects on cardiomyocytes, compared to combinatorial treatments with clinically validated monoclonal antibodies (mAbs). Interestingly, the combination of novel human PD-L1_1 (anti-PD-L1 mAb) and ID-1 (anti-CTLA-4 mAb) induced efficient lysis of triple-negative breast cancer cells and lower cytotoxicity on cardiomyocytes, when co-cultured with lymphocytes, compared to the treatments with Ipilimumab, Atezolizumab or their combination. We further evaluated the pro-inflammatory effects of FDA-approved combinatorial treatments of ICIs, and we found that these combinations, especially regimens including the anti-LAG-3 antibody Relatlimab, promoted an important increase of pro-inflammatory cytokines secretion, such as IL-6, IL-1β, TNF-α, IFNγ and granzyme B.Since combinations of novel mAbs showed a better profile, we decided to fuse the binding moieties derived from LAG-3_1 with those of PD-L1_1 and PD-1_1, generating four novel bi-specific immunomodulatory tribodies. The novel compounds showed binding affinities for their targets comparable to those of validated mAbs (Atezolizumab, Pembrolizumab or Relatlimab) and, more interestingly, an increased ability to induce lymphocyte activation and stronger in vitro tumor cytotoxicity compared to the combination of clinically validated mAbs.In parallel, with the aim of recruiting T cells against cancer cells, we also generated novel tri-specific T-cell engager (TCE) tribodies, able to simultaneously recognize the oncofetal antigen 5T4 on tumor cells, CD3 on T cells, and an IC (PD-L1, PD-1 or LAG-3) by including the scFvs derived from the mentioned novel mAbs. We found that the novel TCEs enhanced cytokine release from hPBMCs, with a strong secretion of IL-2 and IFNγ and showed more potent antitumor effects than the combination of the parental bispecific compound and clinically validated mAbs targeting the same ICs. Notably, the TCE including the anti-PD-L1 moiety induced complete tumor regression in a mouse model.To increase the collection of mAbs specific for ICs, human anti-OX-40 scFvs were identified by phage display selections on hPBMCs and purified receptor and used to generate four novel fully human mAbs targeting this immunostimulatory receptor expressed on immune cells. The novel mAbs were tested for their biological effects on human lymphocytes alone or in co-cultures with tumor cells and enhanced both T cell activation and tumor cell killing, displaying additive effects when combined with Atezolizumab. Finally, a novel human antibody targeting the coagulation cascade, and in particular Factor V, was also generated by using phage display. The novel mAb, called D9, was found able to efficiently bind to Factor V in both its native and activated forms, with nanomolar KD values and to slow down the coagulation by prolonging the activated partial thromboplastin time (aPTT). These features could make this mAb a potential tool for both diagnostic and therapeutic applications.Overall, these findings highlight the potential of the newly generated bi- and tri-specific antibodies to overcome the limitations of
NOVEL MONO-, BI- AND TRI-SPECIFIC IMMUNOMODULATORY ANTIBODIES FOR CANCER THERAPY AND TOOLS FOR OTHER THERAPEUTIC AND DIAGNOSTIC APPLICATIONS / R. Rapuano Lembo ; tutor: N. Zambrano ; co-tutor and supervisor: C. De Lorenzo ;coordinator: D. Pasini. Dipartimento di Oncologia ed Emato-Oncologia, 2025 Dec 19. 37. ciclo, Anno Accademico 2024/2025.
(POST)COLONIAL INTERFERENCES: ETHNICITY, GENDER, AND CLASS IN CONTEMPORARY DANISH LITERATURE ABOUT GREENLAND
During the last two decades, Greenland has progressively become a hot topic within the international public debate, and this same interest has characterized Danish literature – in a variety of forms ranging from autofictional and historical texts to genre-fiction. Kim Leine, Iben Mondrup, and Lotte Inuk stand out as three significant representatives of this wave, also due to their peculiar experience as migrants in Greenland. The present work is concerned with the narratives that have characterized Danish literature and discourses about Greenland since the cultural encounter between former colonizer and colonized and investigates whether the three authors’ ‘internal’ viewpoint has led to literary understandings of Danish-Greenlandic relations that do not necessarily rely on stereotypical stances or on a binary opposition between ‘us’ and ‘them’. Two discourses – both central to postcolonial studies pertaining to the Danish-Greenlandic context – are at the core of my inquiry: 1) the ‘othering’ dynamics that fall under the name of «Arctic orientalism», concept theorized by Kirsten Thisted in the 1990s and 2) the so-called «exceptionalism», i.e., the rhetoric of self-absolution promoted on Danish side with regard to colonial presence and responsibility in Greenland. The (possible) deconstruction of these narratives in a selection of novels by Leine, Mondrup, and Inuk has been analyzed by taking into consideration three different categories of social difference: ethnicity, gender, and class. The joint attention for these aspects – all deeply intertwined with understandings of identity – presupposes an ‘intersectional’ viewpoint on Greenlandic society and how it is represented in these literary works. Although in the course of my research I also refer to the notion of «intersectionality» – first employed by the jurist Kimberlé Crenshaw in an attempt to «contrast the multidimensionality of Black women’s experience with the single-axis analysis that distorts these experiences» (Crenshaw 1989: 139) – I rather tend to employ Ingunn Moser’s adjoining term, «interference». ‘Interference’ points to the fact that «the different processes that construct gender, class and dis/ability not only mechanically sustain and reinforce each other (as the metaphor of intersections suggests), but may also clash, come into conflict and neutralize each other» (Lykke 2010: 84). Moser’s expression, suggesting a certain degree of ‘fuzziness’, thus seems to leave more room for possible inconsistencies.The ‘intersectional’ nature of my work unfolds through the creation of a ‘cumulative’ structure for the last three chapters – preceded by chapter 1, conceived as a historical introduction to the context and genre here scrutinized. While chapter 2 solely revolves around the ethnic dimension, chapter 3 and 4 – respectively focusing on gender and class – are essentially ‘multidimensional’, as they build on concepts and observations made in the previous sections of the dissertation. Thus, the category of ethnicity constantly dialogues with those of gender and class; gender-related matters are also discussed in chapters 2 and 4, although they are perhaps less pervasive than discourses on ‘Danishness’ and ‘Greenlandicness’, at the center of dansk grønlandslitteratur.
(POST)COLONIAL INTERFERENCES: ETHNICITY, GENDER, AND CLASS IN CONTEMPORARY DANISH LITERATURE ABOUT GREENLAND / F. Turri ; tutor: C.C. Storskog ; co-tutor: K. Thisted ; coordinator: L. Pinnavaia. Dipartimento di Lingue, Letterature, Culture e Mediazioni, 2024 Jun 21. 36. ciclo, Anno Accademico 2022/2023.
TOWARDS A BETTER UNDERSTANDING OF THE ROLE OF FATIGUE ON ACL INJURY RISK
The anterior cruciate ligament (ACL) originates on the posterior-superior portion of the medial facet of the lateral femoral condyle and inserts anteriorly and medially to the tibia's intercondylar eminence, preventing the femur from sliding from the tibia anteriorly. Non- contact ACL lesions are one of the most devastating injuries in sports, and they often occur during abrupt changes of direction, like turns or cutting maneuvers and landing movements. ACL injuries during sports movements usually result from the interaction of several extrinsic and intrinsic risk factors, which may change dynamically due to physical fatigue.The overall scope of this dissertation was to better elucidate the effect of physical fatigue on the biomechanical risk to sustain an ACL injury during sports movements. Since fatigue has been defined in several ways, the presented protocols intended to replicate, still considering the laboratory environment limitations, the real sport-like scenario miming different harmful athletic movements (e.g., change of direction, landing from a jump). Furthermore, the procedures aimed to stimulate both the central and peripheral mechanisms responsible for the fatigue onset in an athletic population composed of team-sports athletes from the amatorial to the elite level.The first study evaluated the effect of a 5-minutes continuous shuttle run protocol involving changes of direction on the kinematics of the lower limbs in recreational athletes, showing kinematic changes previously suggested to increase the risk of ACL injury in cutting tasks, such as the substantial reduction of hip and knee flexion, as well as the increase of hip adduction and hip internal rotation.The second project aimed to replicate and ameliorate the first one with the involvement of elite female soccer players, who performed the shuttle run till exhaustion to evaluate both the kinematics and kinetics of the lower limbs. The repetition of turning actions induced alterations in lower limb kinematics in all the tested players and generated changes in lower limb kinetics in 85% of the players. The nature and extent to which lower limbs mechanics were altered throughout the test were highly variable among individuals. Biomechanical modifications mostly occurred in the sagittal plane, but adaptations in the coronal and transverse planes were also detected. A subset of participants showed a drift of pivoting limb kinematics that tightly matches with the known ACL injury mechanism, while other players displayed less definite, minimal, or even opposed behaviors.Furthermore, a systematic review was carried out to explain the associations between cognitive function and ACL injury-related biomechanics and introduce the context for the third project. Five studies out of six found some form of a cognitive-motor relationship, with worse cognitive performance associated with an increased injury risk from the dual-task scenario. At the same time, the remaining study did not identify interactions between cognitive function, secondary tasks, and landing mechanics.The last original project was performed during the visiting period at the Neuromuscular Biomechanics Laboratory of the Montana State University and aimed to assess the combined effect of cognitive performance and physical fatigue during an unanticipated landing task, in order to define the interaction between these potential risk factors. The peak knee flexion angle decreased throughout fatigue progression, demonstrating a potentially harmful movement pattern when the athletes were fatigued, especially at the end of the protocol. In addition, this study highlighted some interesting fatigue effects during unanticipated landing, especially relationships with the participants' baseline cognitive performance.
TOWARDS A BETTER UNDERSTANDING OF THE ROLE OF FATIGUE ON ACL INJURY RISK / F. Bertozzi ; tutor: C. Sforza, C. Galvani, S.M. Monfort ; coordinatore: C. Sforza. Università degli Studi di Milano, 2022 Apr 21. 34. ciclo, Anno Accademico 2021.
FROM A PLACODE PROGENITOR PLATFORM TO A TRIGEMINAL GANGLION ORGANOID: MODELLING SENSORY DEVELOPMENT AND FUNCTION IN VITRO
The trigeminal ganglion (TG) is composed of peripheral sensory neurons which transmitpain, temperature and touch information from the face to the brain. Developmentally, itoriginates from an ectodermal population named pre-placodal ectoderm, which is markedby the expression of the transcription factor SIX1, and which gives rise to most of the facialperipheral nervous system. The intermediate region of the pre-placodal ectoderm lateracquires a trigeminal fate, marked by the expression of PAX3, and thus develops into theTG. The TG is involved in several human pathologies, such as migraine and viral diseases,including the establishment of viral latency in trigeminal sensory neurons by herpesviruses.TG-associated pathologies remain poorly understood in part due to a lack of human,scalable in vitro models of the ganglion. Human pluripotent stem cells can be differentiatedtowards three-dimensional organoids mirroring tissue development, morphology andfunction, but organoids of the TG are yet to be developed. Here, we aim at generating thefirst TG organoid (TGO) and at characterising its development, maturation and function. Bymimicking TG development in vitro, we successfully generated a multipotent placodeprogenitor organoid (PPO) composed almost exclusively of cells expressing the placodalmarker SIX1. To guide PPOs towards an intermediate fate expressing PAX3, we performeda high-content chemical compound screen and identified novel signalling pathwaysimportant for trigeminal development. We thus obtained an organoid enriched in trigeminalprogenitors, which, when cultured over time, developed into trigeminal sensory neurons.Characterisation of TGOs by scRNA-seq and spatial transcriptomics at defineddevelopmental time points highlighted the emergence of distinct sensory neuron and glialsubpopulations, which transcriptionally and spatially resembled in vivo TG cells.Importantly, the sensory neurons in TGOs are functional, as they are able to fire actionpotentials in response to pain and temperature-mimicking stimuli. TGOs thus closelyresemble human TG in vivo in their development, cell composition, and function. Theyconstitute the first human scalable system to investigate TG physiology and pathology, andcould be employed in the future to shed light on diseases such as migraine and viral latencyin the ganglion.
FROM A PLACODE PROGENITOR PLATFORM TO A TRIGEMINAL GANGLION ORGANOID: MODELLING SENSORY DEVELOPMENT AND FUNCTION IN VITRO / E. Conti ; tutor: O. Harschnitz ;internal advisor: J. Davila-Velderrain ;coordinator: D. Pasini. Dipartimento di Oncologia ed Emato-Oncologia, 2025 Dec 16. 37. ciclo, Anno Accademico 2024/2025.
CHARACTERIZATION OF THE EPIGENOMIC PROFILE SHAPING HUMAN FOXP3+ REGULATORY T CELL IDENTITY IN THE TUMOR MICROENVIRONMENT
One of the hallmarks of cancer is the evasion of the anti-tumor immune response. Cancer cells orchestrate several mechanisms to grow unchecked while promoting a suppressive tumor microenvironment (TME). In this context, CD4+ FOXP3+ regulatory T cells (Tregs) have emerged as a major immunosuppressive population sustaining cancer progression. In physiological conditions they mediate immune tolerance and maintain balance in immune responses, controlling their intensity and timing. Moreover, Tregs can acquire specific phenotypes to modulate distinct immunological contexts and regulate tissue homeostasis through mechanisms that extend beyond their canonical immune functions.Tumor-infiltrating Treg cells (TI-Tregs) are frequently found in solid tumors, and their ratio compared to CD8+ effector T cells often correlates with poor patient prognosis. This makes the selective depletion or reprogramming of TI-Tregs a promising strategy in cancer immunotherapy, aiming to enhance anti-tumor immunity without disrupting Treg-dependent homeostasis in healthy tissues. Previous studies have revealed specific transcriptional programmes of TI-Tregs compared to other immune subsets, including Tregs in the blood (PB-Tregs) and normal tissues adjacent to tumors (NAT- Tregs). These findings pave the way for discovering novel TI-Treg-specific targets. However, a deeper molecular profiling of this Treg cell state, including their regulatory landscape shaped by TME cues, is required to develop novel immunotherapy approaches.Here, we focus on epigenomic rewiring as a key process that orchestrates Treg adaptation and the acquisition of a tumor-promoting phenotype. The integration of external signals dynamically alters various layers of epigenomic regulation, mediating the establishment of tailored gene expression programs. In this regard, enhancers and DNA-binding transcription factors (TFs) are pivotal in forming specific regulatory networks.To map the TI-Treg epigenomic profile, CD4+ CD25+ CD127- Tregs were isolated from the blood (PB), tumors, and normal adjacent tissue (NAT) of colorectal cancer (CRC) and non-small cell lung cancer (NSCLC) patients. Chromatin accessibility and different histone modifications were profiled using ATAC-seq and ChIPmentation-seq, respectively, to reconstruct the genome-wide active and repressed chromatin states and to precisely map open chromatin regions with enhancer features. Additionally,5single-cell RNA-seq data were leveraged to discover bidirectionally transcribed noncoding enhancer RNAs (eRNAs), another feature marking active enhancers and delineating the portion of the enhancer region most likely mediating enhancer function and TF binding. TF motif analysis and in silico TF footprinting were employed to assess TF binding dynamics and differential regulatory networks across Treg cell states.We map the comprehensive enhancerome of human Tregs through the integration of multiple epigenomic features assessed in independent omics datasets. This atlas includes 14,158 bona fide active enhancers in TI-Tregs and in normal tissue-Tregs. Notably, our combined multi-omics approach highlights that enhancers comprise a minor fraction of open chromatin regions in Tregs, underscoring the advantage of our approach in robustly annotating active enhancers compared to other accessible regions.We reveal distinct open chromatin landscapes across PB-, NAT-, and TI-Treg cell states, with altered accessibility in cis-regulatory elements including enhancers. Focusing on TI-Treg chromatin rewiring, we find that epigenomic changes at enhancer level reflect transcriptional differences with respect to NAT-Tregs. Moreover, the existing specificities between these two Treg populations, confirmed by our findings, are driven at a much greater extent by enhancers as opposed to promoters. Integration of epigenomic and transcriptomic data uncover different regulatory programs involving coordinated changes in enhancer activity and target
CHARACTERIZATION OF THE EPIGENOMIC PROFILE SHAPING HUMAN FOXP3+ REGULATORY T CELL IDENTITY IN THE TUMOR MICROENVIRONMENT / C. Dossena ; tutor: M. Fakiola ; director: M. Pagani. - L.I.T.A, Via Fratelli Cervi 93, Segrate (MI). Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, 2024 Jan 15. 37. ciclo, Anno Accademico 2023/2024.
SMN DEFICIENCY INDUCES NEUROACTIVE AMINOACIDS DYSMETABOLISM: BRIDGING INSIGHTS FROM PRECLINICAL MODELS TO CLINICAL EVIDENCE IN SMA PATIENTS
ABSTRACTSpinal muscular atrophy (SMA) is a leading genetic cause of infant mortality, driven by homozygous loss of the SMN1 gene and reduced SMN protein. Although disease-modifying therapies such as Nusinersen, Risdiplam, and Onasemnogene abeparvovec have markedly improved survival and motor outcomes, they are not curative, and many patients continue to exhibit residual weakness, metabolic instability, and heterogeneous responses. This underscores the need for SMN-independent therapeutic strategies and robust biomarkers to enable personalised disease management.We investigated how SMN deficiency disrupts amino acid metabolism and neurotransmission across central and peripheral systems, integrating data from SMA mouse models, Drosophila, and patient biofluids. First, we show that excitatory amino acids, including L-glutamate, L-glutamine, D-serine, and D-aspartate, are dysregulated in the CNS and cerebrospinal fluid (CSF) of SMA, compromising glutamatergic neurotransmission—supplementation with D-serine improved motor performance in severe SMA mice, identifying a tractable metabolic target. Second, we discovered a selective deficiency of L-arginine in the spinal cord of SMA mice and in CSF from SMA1 patients, which Nusinersen restored in severe cases. Given L-arginine’s dual role in nitric oxide signalling and nitrogen/energy metabolism, this finding links SMN loss to both synaptic and vascular dysfunction. Third, we identified taurine deficiency in the SMA mice brainstem and CSF of patients, also corrected by Nusinersen in SMA1, nominating an additional candidate for adjunctive therapy.Extending beyond the CNS, systematic profiling in SmnΔ7 mice revealed widespread depletion of L-arginine and L-alanine across skeletal muscle, liver, kidney, and serum, alongside elevated glutamine/glutamate ratios, a systemic signature conserved in Drosophila SMA models. These metabolic changes were accompanied by stereospecific imbalances in D/L amino acids, as well as DNA damage and innate immune activation, establishing links between SMN loss, metabolic stress, and genome instability. Collectively, these studies establish amino acid dysregulation spanning central, peripheral, and stereospecific dimensions as a conserved hallmark of SMA. By integrating cross-species evidence, they highlight amino acids as grounded therapeutic candidates for SMN-independent intervention. This work provides a translational framework for developing biomarker-guided combination strategies to improve long-term outcomes in SMA.
SMN DEFICIENCY INDUCES NEUROACTIVE AMINOACIDS DYSMETABOLISM: BRIDGING INSIGHTS FROM PRECLINICAL MODELS TO CLINICAL EVIDENCE IN SMA PATIENTS / A. Hassan ; supervisor: A. Usiello ;internal supervisor: F. Salvatore ; internal evaluator: E. M. Valente ;external evaluator: M. Bowerman. Dipartimento di Oncologia ed Emato-Oncologia, 2025 Dec 19. 37. ciclo, Anno Accademico 2024/2025.
PATIENT SIMILARITY NETWORKS-BASED METHODS FOR MULTIMODAL DATA INTEGRATION AND CLINICAL OUTCOME PREDICTION
"Artificial Intelligence is significantly contributing to the development of Precision Medicine, a revolutionary approach in healthcare. This new medical paradigm leverages heterogeneous patients' descriptions to enhance disease prevention, diagnosis, prognosis, and treatment.The progress of Precision Medicine significantly relies on the ability to collect comprehensive data from each patient, covering various aspects of their disease. This includes gathering information at different levels to form a complete picture of the pathology, incorporating genomic, environmental, and lifestyle factors. Recent technological advancements have facilitated the ongoing collection of a vast array of diverse health data. This includes clinical data, radiological and digital pathology images, molecular data, and information from wearable devices and smartphones. Such developments have ushered the field of biomedicine into the era of ""Big Data"", presenting unique challenges in data analysis.Among these various data types, multi-omics data has become the gold standard for the in-depth molecular-level characterization of individuals. Multi-omics data involves measuring a comprehensive set of biomolecules that describe a specific biological layer. Examples include DNA sequencing, gene expression levels, DNA methylation, and protein expression. As a result, this data is both multimodal and high-dimensional, offering a more nuanced understanding of biological processes.Network Medicine is an innovative and interdisciplinary field of bioinformatics research that applies the principles of network science to understand and treat diseases. It revolves around the concept that most diseases are not caused by individual genes, but rather by complex interactions within biological networks. In Network Medicine, diseases are viewed as the result of perturbations in these networks. By understanding these network interactions and their degradation, researchers can better understand the multifactorial nature of diseases, therefore providing crucial insights that enable Precision Medicine to develop more targeted and effective prevention, diagnosis, prognosis, and treatment strategies.In the current landscape, Patient Similarity Networks (PSNs) are emerging as a powerful tool for representing patients' clinical and molecular profiles. These networks adapt the principles of Network Medicine, originally applied to biomolecular networks, to patient data. In a PSN, each node represents a patient, and the edges express the degree of similarity between patients' clinical or biomolecular profiles. PSNs have proven particularly effective in key Precision Medicine tasks such as patient subtyping and predicting clinical outcomes through clustering and classification techniques. They offer the benefits of being interpretable and privacy-preserving, and are capable of integrating multimodal data.The growing availability of multi-omics data, combined with the impressive performance of PSN-based models, underscores the need for developing sophisticated methods to integrate multimodal data in constructing PSNs. This approach promises to enhance the effectiveness of PSNs in capturing the nuances of patient profiles, further advancing the capabilities of Precision Medicine.The first contribution of this thesis is an extensive literature review that collects and critically analyses multimodal data fusion methods for the computation of PSNs. Firstly, recognizing that the development of PSNs is highly dependent on the selection of patient similarity measures, we conduct a comprehensive review of the most commonly used and effective similarity metrics, encompassing multiple data types, including categorical, discrete, continuous, and binary. Next, considering the wide and varied range of literature works about multimodal data fusion techniques with numerous taxonomies proposed over the years, we compiled and organized all of them to offer a clear starting po"
PATIENT SIMILARITY NETWORKS-BASED METHODS FOR MULTIMODAL DATA INTEGRATION AND CLINICAL OUTCOME PREDICTION / J. Gliozzo ; supervisor: G. Valentini ; co-supervisor: E. Casiraghi, A. Patak ; coordinator: R. Sassi. Dipartimento di Informatica Giovanni Degli Antoni, 2024 Apr. 36. ciclo, Anno Accademico 2022/2023.
ANALYSIS OF THE DEATH EVENTS, ACCORDING TO THE HISTORICAL, DEMOGRAPHICAL AND TERRITORIAL CONTEXT OF MILAN PEOPLE. DATA FROM THE MILANO SFORZA REGISTERS OF THE DEAD (1452-1801)
The Milano Sforza Registers of the Dead, a corpus of 366 volumes, is one of the oldest mortality registers of any large European city. Established in 1450 by Duke Francesco Sforza, they are an example of disease monitoring and prevention. The information in them reflects death events at the individual levels and includes both socio-demographical context and clinical data (causes of death and description of the illness).The information network that compiled the registration started with the parish elder; it included health officials and ended with the notary at the Health Office who transcribed the information.REDCap (Research Electronic Data Capture), a web-based application developed by Vanderbilt University, allowed the creation of a database to store the data of the registers and ensured their integrity. The data of 1480, a year without plague and war, was taken as an example to show the database structure for the registers. The database is essential to perform statistical analysis based on the data.The registrations of 1480 were used to study the event distribution for different age classes and gender. They were used to study the distribution of the events in different parts of the city and the time trend. Detailed descriptions of the death in foundlings, death for childbirth, violent causes of death, and hospital events are provided.Two plague periods were then considered: 1630 and 1485. Data about 1630 were used to perform the spatiotemporal reconstruction of a famous epidemic, known all over the world thanks to the work of Alessandro Manzoni. Data about 1485 were used to analyze the association of plague symptoms, differences between life and death for plague, and to study the time distribution between symptoms and death. Finally, Ferrario's time series, which was compiled in 1840s, and aggregated information about the number of deaths in each month, was used to find the outliers that can be connected to extraordinary events and to subdivide the series into periods with similar characteristics. With this aim, a R package for the ecological time series was used.
ANALYSIS OF THE DEATH EVENTS, ACCORDING TO THE HISTORICAL, DEMOGRAPHICAL AND TERRITORIAL CONTEXT OF MILAN PEOPLE. DATA FROM THE MILANO SFORZA REGISTERS OF THE DEAD (1452-1801) / E. Luconi ; tutor: E. Biganzoli ; coordinatore: C. La Vecchia. Dipartimento di Scienze Biomediche e Cliniche, 2023. 36. ciclo, Anno Accademico 2023.
DISTRIBUTED COMPUTING BY MOBILE ROBOTS: COMPUTATIONAL POWER AND ALGORITHM DESIGN
The Look-Compute-Move (LCM) model is a theoretical framework widely used to formalize and study distributed systems of mobile autonomous robots, namely swarms of robots.Generally, the LCM model assumes robots are anonymous, indistinguishable, homogeneous, and punctiform entities acting on the Euclidean plane through LCM cycles.Other features may limit robots' power: e.g., mostly, literature considers disoriented robots, i.e., devoid of a global coordinate system, and with limited or absent storage and communication means.Each robot is embedded with the same deterministic algorithm, which is executed at any robot activation within an LCM cycle.Thanks to such an algorithm, the swarm solves a problem; typically, research focuses on problems requiring, for instance, the swarm to arrange in specific geometric patterns, or to move to satisfy some conditions.This thesis encompasses two research topics mainly involved in this field.The first part investigates the computational power of different LCM sub-models, which vary in certain robot features (i.e., memory, communication, visibility, and synchronization). Notably, we study whether different sub-models are able to solve the same set of problems or specific problems (e.g., the Fault Detection problem in fault-prone systems). The second part concerns the design and analysis of algorithms for solving specific formation problems under selected sub-models. Specifically, we provide two algorithms for the Uniform Circle Formation problem, which requires a swarm of n robots—that are luminous (i.e., equipped with constant-size memory and communication means), opaque (i.e., subject to obstructed visibility in the case of collinearity), and asynchronous—to form a regular n-gon. In particular, the first algorithm is O(log n)-time, while the second one is time-optimal.Finally, we constructively prove that a swarm of luminous and sequential robots (i.e., activated one at a time) can solve the Universal Dancing problem, which requires the swarm to form sequences of any patterns, starting from any initial configuration.
DISTRIBUTED COMPUTING BY MOBILE ROBOTS: COMPUTATIONAL POWER AND ALGORITHM DESIGN / C. Feletti ; tutor: B. Palano ; co-tutor: C. Mereghetti ; coordinator: R. Sassi. Dipartimento di Informatica Giovanni Degli Antoni, 2025 Dec 09. 38. ciclo, Anno Accademico 2024/2025.
MICRO- AND NANOPLASTIC: FATE IN THE FRESHWATER ECOSYSTEM, FORMATION THROUGH THE USE OF PLASTIC WATER BOTTLES, AND A MODEL FOR HUMAN HEALTH RISK EVALUATION
Although the degradation of plastics is desirable, the degradation products of plastic materials, micro- and nanoplastics, are of increasing concern. These plastic particles are now emerging pollutants in the environment and are considered potentially hazardous to organisms and humans. This thesis had the scope of giving further insights into the understanding of thesepollutants by addressing fate, generation, detection and effects on human health. In detail, the following questions were posed: How do microplastics distribute in a river ecosystem and does the biotic microplastic pollution reflect that of the abiotic matrices? Can the use of plastic packaging generate plastic particles? How are nanoplastic particleschemically modified during secondary formation, and through which analytical approach can they be identified and quantified? What are the human health effects of inhaled microplastics,and is the 3D structured human airway organoid model suitable for human risk assessment of atmospheric plastic particles?For investigating the occurrence and fate of microplastic in a freshwater ecosystem, two projects were conducted on the Ticino River (North Italy); one on microplastics extracted from multiple matrices (water, sediment, fish and macroinvertebrates) simultaneously sampled along the river and one on microplastic isolated from pellets of an avian freshwater species.The results demonstrated a high complexity of the distribution of microplastics along the river, evidenced by the absence of correlation of concentration between the matrices and along the river, and that a single matrix alone cannot accurately represent the microplastic pollution level of a river ecosystem. Moreover, the spatial variability and the opposing microplastic concentrations in water and sediment further indicates a strong role of the pronounced hydrodynamic conditions of the Ticino River, where microplastics can be deposited, retained and resuspended. Furthermore, using regurgitated pellets and a tied use of different analytical techniques, here µ-FTIR and SEM-EDS, were proven to be a suitable approach for identifying (micro)plastics for avian plastic ingestion studies.The formation of plastic particles from packaging was investigated in two further projects focusing on the release of micro- and nanoplastic particles from drinking water plastic bottles (single-use) under simulated use. The results demonstrate that the bottle lid (made of high-density polyethylene) was the source of the formation of plastic particles, which were also detected on bottlenecks and, therefore, available for human exposure via ingestion. In this context, the modifications of the physical-chemical characteristics of secondary nanoplastic particles were detected and described. High resolution SEM, XPS, SPES and µ-Raman analysis, in combination with concentration steps proved to be suitable for quantifying and identifying nanoplastics in simple matrices like drinking water.The last project was dedicated to an innovative model for the risk evaluation of inhaled and deposited atmospheric micro- and nanoparticles on humans. In this work, 3D human airway organoids were characterised and exposed to synthetic microplastic fibres released by drying textiles in a tumble dryer. The result shows that the presence of nonbiodegradable fibers during the repair phase of a damaged lung epithelium may lead to their inclusion in the repaired tissue with unknown effects on long term perspective. Concluding that human airway organoids are suitable for testing airborne micro-and nanoplastics to determine the potential risks of atmospheric particles in developing adverse pulmonary effects, this thesis contributes to the development of urgently needed human models for assessing the impact of particulatematter pollutants.Covering different aspects of micro- and
MICRO- AND NANOPLASTIC: FATE IN THE FRESHWATER ECOSYSTEM, FORMATION THROUGH THE USE OF PLASTIC WATER BOTTLES, AND A MODEL FOR HUMAN HEALTH RISK EVALUATION / A.s. Winkler ; tutor: D.Rubolini ; co-tutor: P. Tremolada. Università degli Studi di Milano, 2022 Apr 29. 34. ciclo, Anno Accademico 2021.
CATALYTIC-INDEPENDENT FUNCTIONS OF KDM6A IN B CELL MATURATION AND IMMUNE REGULATION
Kdm6a is an X-linked gene encoding a lysine demethylase involved in epigenetic regulation of gene expression. It catalyzes the demethylation of histone H3 lysine-27, counteracting the function of Polycomb Repressive Complex 2 (PRC2). Kdm6a plays a key role in transcriptional activation of genes that control cell differentiation during embryonic development and postnatal life. It is recurrently mutated in several solid and hematological cancers and is responsible for Kabuki syndrome. While Kdm6a has been extensively studied in early hematopoiesis, T-cell development, immunity, and plasma cell differentiation, its role in B-cell maturation and immunity has been largely overlooked. This study aims to elucidate the role of Kdm6a in B cell differentiation and its influence on adaptive B cell immune responses. To achieve this goal, I generated B-cell-specific Kdm6a gene-targeted mice using Cre/loxP recombination technology in vivo. In these conditional mutant mice, Kdm6a inactivation started in pro-B cells and reached completion in pre-B cells and later stages of B-cell lymphopoiesis.Kdm6a deficiency was dispensable for early B-cell differentiation but limited the re-entry of recirculating mature B cells into the bone marrow. Kdm6a-deficient mice exhibited a significant reduction in follicular B-2 B-cells in the spleen, peritoneal cavity lavages, and Peyer’s patches. This defect was linked to reduced expression of the Baff receptor in mutant B cells. Conversely, the fraction and absolute number of splenic marginal zone (MZ) B cells were increased in Kdm6a-deficient mice. Self-replenishing B-1 B cells in the peritoneal cavity remained unaffected by Kdm6a deficiency.Following immunization with T-cell-dependent antigens, B-cell-specific Kdm6a mutant mice exhibited a modest but consistent increase in the fraction of germinal center (GC) B cells in the spleen. A significant increase in GC B cells was also observed in the Peyer’s patches (PP) of Kdm6a-deficient mice. Flow cytometric analyses revealed an increased fraction of Cxcr4hi centroblasts at the expense of Cxcr4lo centrocytes in Kdm6a mutant mice.Kdm6a mutant GC B cells expressed higher transcript levels for genes promoting cell cycle progression, DNA replication, and mitosis. Conversely, genes associated with chemotaxis and trafficking, including Ccr6, Ccr7, Ebi2/Gpr183, Itga4, and Itgb7, were significantly downregulated upon Kdm6a inactivation. B-cell-specific Kdm6a deficiency led to a significant decrease in serum IgA titers, associated with reduced seeding of B cells in PP. Residual Kdm6a-deficient B cells reaching PP were preferentially recruited into the GC reaction. By 18–24 months of age, Kdm6a mutant mice accumulated an atypical population of Cd21loCd23loCd11b+Cd43+ B cells in the spleen, closely resembling aged B cells.Immunoblotting analyses ruled out major changes in global H3K27me3 levels in Kdm6a mutant B cells. Chromatin accessibility studies indicated that the genome-wide map of accessible regions at transcriptional start sites remained largely conserved upon Kdm6a inactivation. These findings align with the highly restricted subset of differentially expressed genes observed when comparing Kdm6a-proficient and -deficient GC and follicular B cells.To assess the significance of Kdm6a’s catalytic function, a comprehensive immunophenotypic analysis was conducted on B cells from germline mutants expressing a catalytically inactive Kdm6a protein. Notably, defects observed in B-cell-specific Kdm6a null mice, including impaired follicular/marginal zone B-cell differentiation, reduced GC B-cell pool size, contraction of the PP B cell pool, and diminished IgA serum titers, were normalized in Kdm6a catalytic-dead animals.In conclusion, this study establishes a non-redundant catalytic-independent role for Kdm6a in regulating follicular and marginal zone B-cell differentiation, homeostasis, and trafficking acros
CATALYTIC-INDEPENDENT FUNCTIONS OF KDM6A IN B CELL MATURATION AND IMMUNE REGULATIO / C. Ranise ; tutor: S. Casola ; co-tutor: C. Vincenzo ; phd coordinator: D. Pasini. Dipartimento di Oncologia ed Emato-Oncologia, 2025 Apr 03. 36. ciclo, Anno Accademico 2023/2024.
IDENTIFICATION OF PHARMACOLOGICAL APPROACHES BLOCKING ALPHA-SYNUCLEIN SYNAPTIC TOXICITY
Misfolding and aggregation of the synaptic protein alpha-synuclein (αsyn) is, at present, considered one of the main drivers of the pathogenesis of Parkinson’s disease (PD). PD is a complex neurodegenerative disorder characterized by progressive loss of DAergic neurons in the substantia nigra pars compacta (SNpc) and deposition of insoluble proteinaceous inclusions containing αsyn, called Lewy Bodies (LB), in different regions of the nervous system. Toxic species of αsyn were demonstrated to affect multiple cellular pathways, mediating synaptic dysfunction even before a dramatic loss of SNpc neurons has occurred. Besides its neurotoxicity towards the DAergic system, αsyn has been recently reported to affect also the corticostriatal glutamatergic signalling, modulating the synaptic levels and activity of NMDA-type glutamate receptors. However, the precise molecular events underlying αsyn synaptic toxicity are still elusive.The synaptic retention of NMDARs was recently demonstrated to be strictly correlated to the interaction with the protein Rabphilin-3A (Rph3A). Interestingly, alterations of Rph3A expression and its interaction with NMDARs at the corticostriatal synapse have been described in advanced PD stages. Besides, a direct αsyn/Rph3A interaction, modulated in presence of LB, has been put forward. Starting from these previous findings, this PhD project is aimed at characterizing Rph3A role in the early stages of PD to identify pharmacological approaches able to slow down αsyn-induced synaptic toxicity. Exploiting different imaging and biochemical approaches, Rph3A was confirmed as a novel αsyn interactor at synaptic sites. In addition, using an in vivo mouse model of αsyn-induced PD, we found that oligomers and fibrils (PFF) of αsyn reduced AMPAR and NMDAR subunits at the striatal excitatory synapse, prior to cause significant DAergic neurodegeneration. At the same time point, αsyn-mice showed decreased striatal dendritic spine density and early motor impairments. Interestingly, I also found that αsyn oligomers and PFF selectively reduced striatal Rph3A postsynaptic levels and Rph3A binding to GluN2A subunit of NMDAR, suggesting Rph3A and Rph3A/αsyn complex as possible mediators of the synaptic dysfunction. Based on these hypothesis, modulatory strategies aimed at restoring Rph3A synaptic levels were firstly tested in an in vitro model of PFF-induced synaptopathy. In particular, a Rph3A/αsyn uncoupling compound (Compound B), identified through a bioinformatic screening, revealed efficacious in blocking spine loss caused by PFF exposure. In the same experimental model, synaptic defects were also prevented by Rph3A overexpression. Based on these in vitro results, these Rph3A-modulatory approaches were then investigated using the αsyn-induced PD mouse model. Notably, chronic intracerebroventricular treatment with Compound B confirmed its efficacy in blocking αsyn-dependent decrease in striatal spine density. Furthermore, striatal delivery of an AAV overexpressing Rph3A resulted sufficient in preventing the early motor impairments found in αsyn-lesioned mice. In conclusion, the results of this PhD thesis demonstrate that Rph3A and Rph3A/αsyn complex significantly contribute to the molecular events underlying early dysfunction of the striatal glutamatergic synapse in PD, therefore representing novel and promising pharmacological targets.
IDENTIFICATION OF PHARMACOLOGICAL APPROACHES BLOCKING ALPHA-SYNUCLEIN SYNAPTIC TOXICITY / E. Ferrari ; tutor: F. Gardoni ; co-tutor: I. Eberini; coordinatore: G. D. Norata. Dipartimento di Scienze Farmacologiche e Biomolecolari, 2021 Dec 09. 34. ciclo, Anno Accademico 2021.
INTEGRATED MOLECULAR INVESTIGATIONS OF MYRF AND SFRP2: UNVEILING REGULATORY MECHANISMS OF ER STRESS AND ECM REMODELLING IN PANCREATIC CANCER
Pancreatic ductal adenocarcinoma (PDAC) stands out as the most prevalent type of pancreatic tumors and the deadliest among common solid malignancies. One of the causes of this high lethality is the significant intratumor cell diversity, highlighting the need for enhanced molecular understanding. Indeed, PDACs are ensembles of three different subtypes with distinct morphology and gene expression programs that coexist in variable proportions in every tumor.Following the identification of transcriptional regulatory networks in the various biotypes, we decided to better characterize two pivotal proteins controlling specific hallmarks of pancreatic cancer biology, like evading growth suppression and activating invasion.First, we focused on the regulation of the myelin regulatory factor (MYRF), a poorly characterized transcription factor. Previous studies conducted in our laboratory demonstrated that MYRF, an endoplasmic reticulum (ER) transmembrane protein, maintains the homeostasis in the ER of pancreatic secretory glandular cells. After trimerization and self-cleavage, MYRF releases the N-terminal fragment which translocates into the nucleus to exert its transcriptional role by regulating the expression of genes encoding secretory proteins. However, the role of C-terminal fragment which remains inserted in the ER is unclear. Since the regulation of MYRF release from the ER membrane is still unknown, our hypothesis is that this fragment may interact with proteins in the ER lumen and regulate MYRF trimerization and cleavage. Using enzyme-catalyzed proximity labeling followed by biotin pull-down and mass spectrometry, we identified MYRF proximal interactors in the ER lumen. Currently, we were able to obtain a first validation of MYRF interaction with UGGT1, which could provide quality control for MYRF folding.Furthermore, in order to investigate the role of MYRF C-terminal fragment in the maintenance of ER functionality, we generated clonal cell lines lacking one of the two critical MYRF functional domains – DNA binding or transmembrane domains - by CRISPR/Cas9-mediated in-frame deletion. The analysis of the gene expression profile of MYRF domain-specific mutants revealed that MYRF has a structural functionality in the ER that is independent of its transcriptional activity.Additionally, we focused on the role of SFRP2 as a potential regulator influencing the characteristics of PDAC transitional cells. One of the key aspects of transitional cancer cells is the ability to contribute to the extracellular matrix (ECM) remodeling: we observed that SFRP2 led to a noteworthy augmentation in this process, along with increased migratory and invasive capabilities of PDAC cells. We also demonstrated that the abundance of ECM components contributes to different tissue density and rigidity, leading cells to mount a mechanoresponsive transcription program.This work could provide i) a mechanistic understanding of MYRF in the regulation of ER activity. Considering the impact of ER stress on cancer cell fitness and drug-induced toxicity, the possibility to influence ER regulation by modulating MYRF trimerization and cleavage by targeting UGGT1 may provide therapeutic opportunities; ii) a better characterization of SFRP2 and its functional effects on PDAC transitional cells, improving anti-cancer therapies.
INTEGRATED MOLECULAR INVESTIGATIONS OF MYRF AND SFRP2: UNVEILING REGULATORY MECHANISMS OF ER STRESS AND ECM REMODELLING IN PANCREATIC CANCER / F. Arco ; tutor: G. Natoli ; co-tutor: D. Pasini ; phd coordinator: S. Minucci. Dipartimento di Scienze della Salute, 2024 Apr 11. 35. ciclo, Anno Accademico 2023.
PRECLINICAL EVALUATION OF A NOVEL NFIX INHIBITOR FOR MUSCULAR DYSTROPHIES TREATMENT
Muscular Dystrophies (MDs) are heterogeneous disorders, characterized by progressive skeletal muscle wasting that, in most severe variants, cause wheelchair dependency and premature death due to respiratory failure and cardiac impairments. The best therapeuticavenue for MDs involves corrective approaches such as cell and gene therapy, along with secondary therapies to slow down disease progression and preserve musclestructure. Working in this direction, in 2017 it has been demonstrated that the inhibition of the transcription factor Nuclear Factor 1 X (Nfix) induces histological and functional improvements of the disease by delaying muscle degeneration/regeneration loops, decreasing inflammatory and fibrotic processes, and shifting myofibers towards a more oxidative and damage-resistant phenotype. Moreover, Nfix specific deletion in macrophages delays muscle wasting and the deposition of fibrosis, reinforcing the therapeutic potential. Considering that MEK/ERK pathway induces Nfix expression, Nfix pharmacological inhibition was tested by repurposing FDA-approved MEK- inhibitors. These drugs decreased Nfix protein levels, but the non-specificity oftreatment resulted in ectopic calcification, partially reduced by antioxidant supplementation. Hence, the finding of a specific Nfix inhibitor has become the central scope of this PhD project. The 3D structure of the Nfix DNA Binding Domain (Nfix(DBD)) was recently resolved, illustrating a novel MH1-like fold and key features for function, including a Zn²⁺ coordination site, dimeric assembly, and DNA-binding specificity. To find molecules capable to bind Nfix, we screened a library of fluorinated compounds using ¹⁹F-NMR spectroscopy, a method widely used in drug discovery, using Nfix(DBD) as a bait. We identified ARN2011 as a promising molecule and, after confirmation of absence of toxicity, we tested its ability in inhibiting Nfix in several cell types. We evaluated ARN2011 efficacy in muscle stem cell, but also fibroblasts and macrophages extracted from murine, rat and human muscles, in wild-type and dystrophic conditions. We displayed that ARN2011 is capable to inhibit Nfix transcriptional regulation but also its ability to control myoblast fusion and macrophage switch from pro-inflammatory to anti-inflammatory phenotype. For what regards the compound specificity, we exploited RNA-Seq analysis on treated myoblasts, comparing wild-type and Nfix null cells, which suggested few off-target events after treatment. Moreover, these data displayed some new insights into Nfix inhibition on cell proliferation, differentiation, metabolism and signaling. Besides, we also tested ARN2011 on murine and human iPS-derived cardiomyocytes revealing its ability to increase myocytes beating rate, in line with what previously published in Nfix null cardiomyocytes. Moreover, ARN2011 was tested on healthy and dystrophic human iPS-derived neuromuscular organoids, confirming its capacity to inhibit Nfix and ameliorate organoid phenotype in terms of histological and contractile abilities. Furthermore, we assessed ARN2011 action in vivo. The proof of concept was performed by intramuscular injection in dystrophic mice models, focusing on metrics analysis of skeletal muscle morphology, such as deposition of fibrosis, inflammatory infiltrates and cross-sectional area of the myofibers. Results displayed Nfix inhibition, followed by decreased inflammatory infiltrates and fibrotic deposit, along with slowdown in muscle regeneration and a switch towards a slow-twitching and oxidative muscle phenotype. ARN2011 systemic evaluation was then performed both intravenously and intraperitoneally, but the correct posology is still to elucidate, considering that preliminary treatments decreased Nfix expression randomly in different muscles, independently from route of administration and timing of inoculation. Besides, some work was performed to elucidate ARN2011 mechanis
PRECLINICAL EVALUATION OF A NOVEL NFIX INHIBITOR FOR MUSCULAR DYSTROPHIES TREATMENT / R. Epis ; supervisore: G. Messina; coordinatore: S. Ricagno. - Milano. Dipartimento di Bioscienze, 2025 Dec 09. 38. ciclo, Anno Accademico 2024/2025.
IL TRATTAMENTO DEI DATI SANITARI AI SENSI DELREGOLAMENTO UE/2016/679
The thesis addresses the issue of health data processing in the light of Regulation 2016/679/EU.Specifically, the first part is dedicated to analyzing the particular regime that the European legislatorhas chosen to introduce, starting from the definition of the health data. After analyzing the legal baseslegitimized for the processing, we investigate how the European legislator has imposed a generalprohibition of processing, except in some instances. In this way, European Member States are free tomaintain or introduce additional conditions while respecting their legal traditions. These exceptionsdemonstrate how the legitimate processing of health-related data represents, in most cases, anessential resource for the benefit not only of the data subject but also of the entire community. Justthink of the processing of health data for scientific research purposes and the impact of the circulationof information and their combination on the development and study of new therapies and scientificadvancement in general. From this point of view, the work's leitmotif is represented by the balancebetween the right to health and the right to data protection. After having illustrated the main doctrinaland jurisprudential theses on the right to health, the analysis focuses on the complex relationshipbetween the right to health and the right to data protection in the context of the tragic emergencycontext caused by the worldwide spread of Covid-19. To this end, starting from the emergencylegislation on the protection of health data in force for the duration of the emergency, it was decidedto address the delicate issue of balancing the two rights, both fundamental. It considers howtechnology, through the introduction of contact tracing apps and the European green digitalcertification, has made a significant contribution to countering the spread of the infection, using datafor a specific public interest. The survey profile that accompanied the entire work concerns the searchfor a balance between the right to health, on the one hand, and the protection of health data, on theother. The study aims to demonstrate how the European legislator himself pays particular attention tothe balance between the two interests, drawing inspiration from the criteria of proportionality andreasonableness and rejecting any logic based on either or the right. For this reason, the same legislatorhas conceived the right to data protection as a flexible right, able to adapt to the purposes pursuedthrough the processing of health data.
IL TRATTAMENTO DEI DATI SANITARI AI SENSI DELREGOLAMENTO UE/2016/679 / V. Danzi ; tutor: G. Carullo ; coordinatore: F. Biondi. Università degli Studi di Milano, 2022 Mar 18. 34. ciclo, Anno Accademico 2021. [10.13130/danzi-vania_phd2022-03-18].
NANOTECHNOLOGICAL APPROACHES FOR CANCER DIAGNOSIS, TREATMENT AND MANAGEMENT: IMPLEMENTATION OF FERRITIN PROTEIN NANOPARTICLES FOR TUMOR TARGETING
Despite the efforts performed in the field of cancer research, data concerning the incidence of this pathology in North America and Europe reveal that tumors still represent the second cause of death. In fact, the effectiveness of many drugs is often seriously affected by lack of specificity in hitting the therapeutic target, determining the onset of toxicity issues. In this context, Nanomedicine aims to make a decisive contribution. Nanoparticle (NPs)-based drug delivery systems can enhance the physicochemical properties of a wide variety of drugs used in oncology to limit off-site side effects and improve their therapeutic efficacy, increasing drug accumulation in target tissue. Thus, through the development of nanocarriers capable of selectively recognizing and interacting with tumor cells and/or tissues, potential groundbreaking applications in the diagnosis, treatment and prevention of cancer could become feasible. In my PhD project, I exploited the unique features of a NPs based on recombinant heavy-chain ferritin nanocages (HFn). HFn nanocages represent a promising protein- based class of NPs for drug delivery widely investigated by virtue of (1) their natural tumor homing and limited accumulation in off-target organs which leads to reduced off-target toxicity; (2) their biocompatibility; (3) their structure that allows the encapsulation of different types of drugs or fluorescent tracers; (4) their versatility to be modified at the surface both exploiting chemical and genetical strategies. My work has been divided into three main studies aimed at implementing innovative strategies based on HFn nanocages both for diagnostic and therapeutic approaches in oncology . In Study 1, I presented the exploitation of HFn nanocages as nanodevices for fluorescence image-guided surgery (FGS). This technique is gaining interest due to its potential to improve tumor margin visualization and real-time identification of tumor deposits, maximizing the benefits for patients. However, no tumor-targeted probe effectively capable of localizing tumor tissue is available to date. For this reason, we proposed a HFn-based nanotracer for the delivery of indocyanine green (ICG), which is the main fluorescent dye currently used in clinics for FGS. A novel variant of HFn was developed, extensively characterized in vitro and loaded with ICG for in vivo experiments demonstrating potentiality as tumor-targeted nanotracers for the identification of tumors. 3 In Study 2, HFn nanocages were exploited for the encapsulation of a chemotherapeutic drug, doxorubicin (DOX) obtaining FerOX nanoformulation. FerOX has been investigated by virtue of its natural tumor homing and limited accumulation in off- target organs which leads to reduced off-target toxicity of drugs herein incapsulated. In particular, the object of this study was to assess the efficacy of FerOX as an alternative formulation capable of preserving the proliferative potential of human lymphocytes and the potential impact in the generation of an adaptative immune response after treatment. In Study 3, we focused on the use of HFn nanocages conjugated to the monoclonal anti-HER2 antibody trastuzumab (TZ). By exploiting the ability of HFn to cross the blood brain barrier (BBB), we achieved the delivery of TZ as a prophylactic treatment to tackle brain metastasis in HER2-positive advanced breast cancer.
NANOTECHNOLOGICAL APPROACHES FOR CANCER DIAGNOSIS, TREATMENT AND MANAGEMENT: IMPLEMENTATION OF FERRITIN PROTEIN NANOPARTICLES FOR TUMOR TARGETING / M. Sevieri ; tutor: S. Mazzucchelli ; coordinator: C. Sforza. Dipartimento di Scienze Biomediche e Cliniche, 2024 May 21. 36. ciclo, Anno Accademico 2023.
META LEARNING IN PROCESS MINING: TOWARD A SYSTEMATIC APPROACH TO DESIGN DATA ANALYTICS PIPELINES WITH EVENT LOGS
With the democratization of computational resources, organizations pay great attention to recording the execution of internal procedures to improve the quality of their services. Modern information systems track and record in-depth data regarding activities performed within their business processes. The event data describes the actual process performance along with several possible attributes. Process mining stands as a set of techniques that leverage insights from event data. With that, organizations can employ process mining-based approaches to understand the processes' behavior, increase the value of services, save resources and improve execution time. Given the multitude of tasks within process mining and the plethora of algorithms and solutions, deciding which methods to apply is a complex effort. Notwithstanding that process mining techniques have now achieved the maturity level to cover the entire stack of the data science pipeline, from raw data to decisions. On the one hand, stakeholders detain business and domain knowledge. On the other hand, they often lack the technical expertise to guide choices. Moreover, many tasks require the application of a combination of several steps, i.e., a pipeline. Designing a suitable pipeline becomes then a complex task, enhanced by the fact that domain experts and technical experts are often not the same people.In this thesis, we propose a task-agnostic framework to automate the design of process mining pipelines. Considering that there is no optimal pipeline for every observable phenomenon, we start from the hypothesis that process behavior might indicate which steps or algorithms are better suited. For that, we rely on a meta-learning approach that maps the relationships between event data and suitable solutions. The application of the proposed framework generates two main contributions. First, given a business process (event log) and a task (e.g., process discovery, trace clustering, anomaly detection), a user can retrieve a pipeline recommendation that best matches the underlying process behavior. The second byproduct of the framework is a systematic mapping of the relationship between event log characteristics and optimal pipelines. This mapping provides experts and data analysts with a solid foundation to better understand the task at hand. That is an enlightenment of the correlation between the problem space (event data), algorithm space (process mining pipelines), and performance space (quality criteria).We instantiated the framework in three different process tasks. Results indicate that indeed there is a relationship between the different spaces since guided recommendations overcome the current baselines. Therefore, showing the importance of investigating guided solutions and that mapping the spaces can be of interest to organizations. Moreover, we investigate which process features are most decisive for each problem. The presented solution is also suitable for users of different knowledge levels. When applying the framework, an inexperienced user has data-based pipeline recommendations whereas an expert is provided with a quantitative mapping that can be used to leverage the knowledge regarding the process task.
META LEARNING IN PROCESS MINING: TOWARD A SYSTEMATIC APPROACH TO DESIGN DATA ANALYTICS PIPELINES WITH EVENT LOGS / G. Marques Tavares ; tutor: P. Ceravolo ; coordinatore: R. Sassi. Dipartimento di Informatica Giovanni Degli Antoni, 2023 Apr 27. 35. ciclo, Anno Accademico 2022.
UNDERSTANDING THE MOLECULAR MECHANISM LINKING ATM TO METABOLIC REGULATION.
Ataxia Telangiectasia (A-T) is a pleiotropic autosomal recessive disease characterized by progressive neurodegeneration, immunodeficiency, sensitivity to ionizing radiation and premature aging. The molecular basis of A-T is loss of function of the gene encoding A-T-mutated (ATM), a serine/threonine kinase that acts as a master regulator of the DNA damage response, leaving A-T patients highly predisposed to cancers. In addition to its classical role in orchestrating the DNA damage response, recent evidence reports a role for ATM in cellular metabolism, oxidative stress response and insulin signaling. However, it is unclear how defective ATM-dependent response to oxidative stress contributes to A-T clinical manifestations. Here we show that ATM activates glycolysis upon oxidative stress and that its loss leads to impairment of glycolysis and impairment of metabolism in A-T cells. In addition, we identified a metabolic regulator as a major driver of glycolysis impairment and poor cell growth in A-T tissues. Respectively, the downregulation and inactivation of this metabolic regulator in A-T cells restored normal glycolysis, reduced metabolites accumulation and improved A-T cells survival and proliferation. Taken together, our results shed light on a new role of ATM in activating glycolysis and cellular metabolism upon oxidative stress and help to identify a metabolic checkpoint activated in ATM-deficient cells to suppress cellular anabolic pathways and cellular growth. Our study also identified a potential therapeutical target that exerts a synthetic survival effect upon downregulation, to slow down the progression and symptoms of A-T disease.
UNDERSTANDING THE MOLECULAR MECHANISM LINKING ATM TO METABOLIC REGULATION / C. El Kharef ; tutor: M. Vinciguerra; internal Advidor: K. Havas ; tutor: V. Costanzo ; phd coordinator: S. Minucci. Dipartimento di Oncologia ed Emato-Oncologia, 2023 Apr 13. 34. ciclo, Anno Accademico 2022.
COVID-VACCINES IN PREGNANCY: MATERNAL AND NEONATAL RESPONSE OVER THE FIRST 9 MONTHS AFTER DELIVERY
AbstractThis doctoral research explores the impact of maternal SARS-CoV-2 mRNA vaccination during pregnancy, focusing on its efficacy in generating immune responses, transplacental antibody transfer, and the subsequent immunity conferred to newborns. The study aims to address gaps in understanding the duration and effectiveness of maternal-derived neonatal immunity while assessing the roles of breastfeeding, maternal booster doses, and postnatal infections in shaping antibody kinetics over a 9-month postpartum period.BackgroundThe emergence of SARS-CoV-2 created significant health challenges, especially for pregnant women, who are at higher risk of severe complications compared to non-pregnant individuals. Despite initial vaccine hesitancy stemming from the exclusion of pregnant women in clinical trials, subsequent observational studies have confirmed the safety and efficacy of mRNA vaccines in this population. Vaccination not only reduces maternal morbidity but also leverages transplacental antibody transfer to provide passive immunity to neonates, a crucial protection mechanism during their immunologically immature early life.MethodologyThis prospective observational study recruited 98 pregnant women who received at least one dose of an mRNA COVID-19 vaccine during different trimesters. Maternal and neonatal antibody levels were assessed at birth (T0), and at 3, 6, and 9 months postpartum (T1–T3). Blood samples were analyzed using chemiluminescent microparticle immunoassay (CMIA) to quantify IgG antibodies against SARS-CoV-2's spike protein. Antibody kinetics were studied in relation to vaccination timing, breastfeeding practices, and postnatal infections. Participants were monitored for vaccine-related adverse events, SARS-CoV-2 exposure, and health outcomes.FindingsMaternal vaccination induced robust immune responses, with efficient transplacental transfer of IgG antibodies observed. The study highlights several critical insights:1. Timing of Vaccination: Antibody levels in both mothers and newborns were significantly higher when vaccination occurred in the second or third trimester. Third-trimester vaccinations resulted in the highest antibody titers in neonates, emphasizing the importance of late-pregnancy immunization.2. Antibody Durability: Neonatal antibody levels peaked at birth and declined significantly over the first 9 months. However, antibodies were detectable in most infants at 9 months, indicating prolonged, albeit waning, immunity.3. Impact of Breastfeeding: Breastfed infants exhibited a slower decline in antibody levels compared to formula-fed or mixed-fed infants, underscoring the complementary protective role of breast milk.4. Role of Booster Doses: Booster doses administered postpartum in mothers significantly increased maternal antibody titers, enhancing neonatal immunity when breastfeeding was continued.5. Postnatal Infections: Neonates exposed to SARS-CoV-2 showed a slower decline in antibody levels, suggesting an active adaptive immune response even in early infancy.Safety and Clinical OutcomesThe study confirmed the safety of maternal vaccination, with no severe adverse effects reported among participants. Neonates exhibited good health outcomes, with no severe COVID-19 cases or hospitalizations observed. These findings reinforce the benefits of vaccination in preventing severe disease in both mothers and infants.DiscussionThe study contributes valuable insights into optimizing maternal immunization strategies. It validates the efficacy of mRNA vaccines in eliciting maternal immune responses and facilitating efficient antibody transfer. Furthermore, it highlights the synergistic effects of vaccination timing, breastfeeding, and booster doses in enhancing neonatal immunity.Key implications include the necessity of late-trimester vaccinations to maximize antibody transfer and the potential of booster doses to sustain ma
COVID-VACCINES IN PREGNANCY: MATERNAL AND NEONATAL RESPONSE OVER THE FIRST 9 MONTHS AFTER DELIVERY / A. Proto ; relatore: G. C. Marchetti ; correlatore: S. Martinelli. Dipartimento di Scienze della Salute, 2025 Feb 28. 37. ciclo, Anno Accademico 2023/2024.
POST-2003 IRAQI FICTION: VOICE, AUDIENCES, AND NARRATIVE AUTHORITY
AbstractThe thesis addresses the non-realist trend of Iraqi fiction written after the US occupation of 2003. In particular, it focuses on the construction and characterisation of non-mimetic voices in this production. Within the frame of post-classical narratology, the thesis asks how the thematic and poetic attention to the traumas of war and migration runs parallel with the construction of narrating-‘I’s that exceed the conventional limits of first-person narration. Such overextended ‘I’s are conceived of as technical supplements, miniature narratives that complement and counterpoint the storyworld – a concept the thesis summarises with the Arabic manṭiq ‘point of articulation’. The thesis is divided into two sections. The first discusses texts in which non-fictional narrative paradigms are unsettled by the challenges of the post-war conjuncture: in particular, it addresses fantastic elaborations of the childhood story (Chapter One) and the ‘asylum story’ demanded of asylum seekers (Chapter Two). The second section is devoted to downright unnatural, anti-mimetic narrative forms. It focuses on two tropes of what has been labelled the Iraqi Gothic: the dead narrator (chapter Three), and the disembodied voice (chapter Four). The analysis of these four paradigmatic narrators of post-2003 Iraqi fiction (the child, the asylum seeker, the dead, and the abstract first-person voice) allows the thesis to address, along with narratological problems, issues relating to the ethics of narration in a traumatic context. Furthermore, the focus on voice involves a questioning of the link between narrative and experience in Iraq.
POST-2003 IRAQI FICTION: VOICE, AUDIENCES, AND NARRATIVE AUTHORITY / F. Pozzoli ; tutor: L. Osti, F. Caiani ; coordinatrice: M. V. Calvi. Dipartimento di Lingue, Letterature, Culture e Mediazioni, 2023 Apr 27. 35. ciclo, Anno Accademico 2022.
P4HB TARGETING HALTS TUMOR GROWTH AND RE-SENSITIZES RESISTANT MELANOMA CELLS TO DABRAFENIB AND TRAMETINIB STANDARD-OF-CARE THERAPY VIA IRE1-⍺ ACTIVATION AND AKT DOWNMODULATION
BRAF-mutant melanoma patients account for approximately 50% of all cutaneous melanoma cases. The overall survival rate of metastatic melanoma patients was dramatically improved by the introduction of targeted therapy (BRAF and MEK inhibitors) and immune checkpoints inhibitors (anti-PD1, anti-CTLA4). However, a significant proportion of BRAF-mutant patients develop secondary resistance to targeted therapies, while others exhibit even primary resistance. To either prevent or overcome resistance in BRAF-mutant melanoma patients, new combinations are now being tested, exploiting triple-agent combinations (targeted therapy with immunotherapy), but with unsatisfactory results. Additionally, the use of immunotherapy in neo-adjuvant conditions, has shown promise, being still under investigation. Another potential therapeutic approach involves re-sensitizing resistant melanoma cells to targeted therapies. However, clinical trials exploring this strategy in patients with BRAF/MEK inhibitor resistance are currently limited. Hence, identifying new vulnerabilities in resistant melanoma cells, as well as to establish novel strategies to re-sensitize resistant melanoma cells to targeted therapies is crucial to improving patient outcomes.Herein, I carried out a functional shRNA-based in vivo screen with a library targeting 195 actionable genes (FDAome library), where each gene is linked to specific FDA-approved drugs that can be repurposed for melanoma treatment. A375 parental cells and A375 cells resistant to dabrafenib and trametinib (A375-DT) were used as models to unveil new vulnerabilities. FDAome screen highlighted 37 genes essential for the in vivo tumor growth of DT-resistant cells. I validated two candidates, namely CREB-binding protein (CREBBP) and Prolyl 4-Hydroxylase Subunit Beta (P4HB), whose inhibition reduced tumor growth in vivo and increased overall survival of A375-DT but not of A375 parental cells, suggesting a putative point of intervention in resistant tumors. Additionally, I demonstrated that silencing CREBBP and P4HB reduced the migratory capacity of A375-DT cells but not A375 cells, further corroborating our in vivo findings. I further validated the in vitro results in a different cell line, namely SKMEL-28-DT and in drug resistant patient-derived primary cultures. To further investigate the therapeutic potential of targeting CREBBP and P4HB, I tested the efficacy of CCS1477 and PACMA31, respectively, as monotherapies and in combination with dabrafenib and trametinib. Both compounds effectively inhibited tumor growth in vivo. Notably, PACMA31 demonstrated the ability to re-sensitize resistant tumors to DT. Furthermore, PACMA31 demonstrated significant efficacy in a syngeneic D4M-DT melanoma model, leading to dramatic tumor regression. Mechanistically, I demonstrated that P4HB silencing had an impact on cellular homeostasis, by increasing accumulation of misfolded proteins and activating IRE1⍺ signalling pathway as part of the unfolded proteins response. This resulted in partial cellular death and autophagy induction. Additionally, P4HB knockdown and subsequent ER stress increment reduced AKT phosphorylation, generating a window of opportunity to re-establish sensitivity to dabrafenib and trametinib treatment.
P4HB TARGETING HALTS TUMOR GROWTH AND RE-SENSITIZES RESISTANT MELANOMA CELLS TO DABRAFENIB AND TRAMETINIB STANDARD-OF-CARE THERAPY VIA IRE1-⍺ ACTIVATION AND AKT DOWNMODULATION / A. Zocchi ; tutor: L. Lanfranconeinternal advisor: P. Ferruccicoordinator: D. Pasini. Università degli Studi di Milano, 2025 Jan 21. 36. ciclo
EVALUATION OF THE GM1 OLIGOSACCHARIDE ROLE IN NEURONAL DIFFERENTIATION
GM1 is a mono-sialo ganglioside with amphiphilic character due to the presence of a hydrophobic group, ceramide, and a hydrophilic head (oligosaccharide chain). GM1 represents one of the most important modulator in the nervous system where it is involve in maturations of neurons, differentiation, increase responses to neurotrophic factors, protection against neuronal death and reduction brain damage. The effects of GM1 are known in vitro and in vivo, but the molecular mechanism of action underlying the GM1 properties is unknown. The present work aims to analyze the mechanism of action of GM1, and in particular to demonstrate that the effects of this ganglioside are attributable to the action of its oligosaccharide portion (OligoGM1) and not to the entire molecule. To reach our purpose we used mouse neuroblastoma cell line Neuro2a (N2a). Like GM1, OligoGM1 promotes neurodifferentiation by increasing both neurite elongation and the expression of neurofilament proteins in N2a cell. A similar effect was obtained with the use of fucosyl-OligoGM1 but not with the administration of asialo-OligoGM1, OligoGM2, OligoGM3, sialic acid or galactose (single components of Oligo GM1). OligoGM1, in N2a cells, activates ERK1/2 pathway binding to the NGF specific receptor TrkA present on the cell surface. To study this mechanism of action we used tritium labeled derivative of OligoGM1. The activator for GM1 mediated functions (differentiation and protection) is the interaction between OligoGM1 and TrkA. This was established with the use of a TrkA inhibition. With a bioinformatics study it was established that OligoGM1 inserts in a pocket of the TrkA-NGF complex. An increase in energy associated to the complex TrkA-NGF-OligoGM1 indicates greater stability of intermolecular interactions.All the results lead to the conclusion that the bioactive portion of GM1, in neuronal differentiation and protection, is represented by its hydrophilic chain (OligoGM1). These conclusions open up new perspectives on the therapeutic use of gangliosides.
EVALUATION OF THE GM1 OLIGOSACCHARIDE ROLE IN NEURONAL DIFFERENTIATION / P. Fato ; tutor: L. Mauri ; coordinatore: A. Prinetti. Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, 2020 Dec 09. 33. ciclo, Anno Accademico 2020. [10.13130/fato-pamela_phd2020-12-09].
AFROBEATS ON SOCIAL MEDIA: TRANSNATIONAL FLOWS, INTERCULTURAL FORMATIONS, AND IDENTITY CONSTRUCTION
ABSTRACTOver the last few years Afrobeats, a contemporary African music from, has steadily been gaining recognition in the global popular culture space. It is not uncommon to see videos of young people dancing to Afrobeats songs circulating on the internet. With the advent of social media platforms like TikTok and their participatory practices, this phenomenon has increased exponentially. This study explores the transnational flow of, and engagement with, Afrobeats within the global sphere. By focusing on TikTok and Youtube as a social mediascape, this study examines the influence of the intersection of social media practices and Afrobeats on digital socio-cultural processes. Guided by the overall research question: how are digital Afrobeats cultures shaping social interaction on social media?, this research combines a number of techniques and methods to understand Afrobeats’ influence on socio-cultural processes. It primarily employs digital methods techniques, using software for collecting and visualising data from TikTok and YouTube. These techniques and tools are combined with digital ethnography, and expert interviews. The study finds that through creative choreographing, and adoption of networked practices TikTokers create a transnational network of Afrobeats content which increases the potential of Afrobeats content, and by extension Afrobeats music, to be encountered across the world. Successful Afrobeats creators on the platform are shaping global popular culture beyond the platform. Furthermore, Afrobeats - a seemingly nonpolitical music form, together with digital platform affordances, is mediating the congregation of transnational and intercultural collectives based on shared socio-political history and experiences. This is achieved through the evocation of collective memories tied to revolutionary and political music of musicians like Bob Marley and Fela Kuti. Finally, this study finds that there is an emergence of a global shift in self-presentation strategies of diasporic Africans from Blackness to Africanness. This shift is tied to the longing of a homeland, and the pride associated with the success of Afrobeats.
AFROBEATS ON SOCIAL MEDIA: TRANSNATIONAL FLOWS, INTERCULTURAL FORMATIONS, AND IDENTITY CONSTRUCTION / B.a.r. Jeduah ; supervisor: P. Rebughini, reviewers: M. Krings, Prof. James Yeku ; coordinator: P. Rebughini. - Milan. Università degli Studi di Milano, 2024 Jul 16. 36. ciclo
USING B-STRAND AND B-HAIRPIN PEPTIDOMIMETICS INDUCED BY CONSTRAINED AMINO ACIDS TO MODULATE THE AGGREGATION OF THE MICROTUBULE ASSOCIATED PROTEIN TAU: DESIGN, SYNTHESIS AND EVALUATION
Tauopathies refer to neurodegenerative disorders characterized by the presence of abnormal aggregates of Tau protein, leading to the formation of neurofibrillary tangles (NFTs) within neuronal cells. Tau, a protein associated with microtubules (MTs), plays a crucial role in regulating the dynamics of MTs and facilitating proper axonal transport in neurons. Alzheimer's disease (AD) is characterized by abnormal protein-protein interactions (PPIs) which lead to the accumulation of β-amyloid plaques and hyperphosphorylated Tau (P-tau) NFTs. Multiple evidence has suggested that the interplay between Tau and Aβ contribute to worsening the disease since the aggregation of one protein can trigger the aggregation of the other. Within this thesis, we have explored, for the first time, the possibility of creating peptidomimetics capable of stabilizing either a β-hairpin or an extended structure to modulate Tau protein aggregation. This work focuses on three distinct peptidomimetic units with the goal of highlighting the significant role of secondary structure in the selective regulation of both physiological and pathological PPIs of Tau. The utilization of a previously developed piperidine-pyrroline β-turn mimic has led to highly effective peptidomimetics that act as dual inhibitors targeting both Tau and Aβ1-42 proteins but show no activity on the other amyloid protein hIAPP. These compounds were designed either on Tau PPIs, by mimicking its self-interaction in NFTs or its physiological interaction with MTs; or on small sequences of the chaperone protein Hsp90, which is a physiological Tau aggregation inhibitor: Through various analyses, including proteolytic stability, ThT fluorescence, cell viability (LDH and MTT), and live cell imaging FDAP assays, we have demonstrated the critical role of pre-organized hairpin structures in achieving potent activity, selectivity, and stability. Among the designed compounds, the one mimicking Hsp90-Tau- interaction showed the highest potency, and NMR titration with 15N-Tau protein provided valuable insights into the possible mechanism of action of Hsp90 on Tau.Then, the employment of the non-natural β2,2-isoxazoline unit into a peptide sequence was used to stabilize two different secondary structures, either in β-hairpin or extended. These structures were confirmed through a combination of CD, IR, and NMR analyses in aqueous solvents. The stabilized hairpin structure was used to develop our most potent inhibitor of Tau aggregation by mimicking Hsp90 protein. The extended structure bearing the PHF6 and PHF6* sequences (both involved in the physiological and pathological processes of Tau), was used to prove that it is the exposure of these sequences on Tau protein the event triggering its aggregation. As a counter-test, the hairpin conformation with the same sequences, did not induce any pathological aggregation. Finally, the development of new difluorinated triazoles and their incorporation into existing peptide inhibitors of Tau, led to enhance their effectiveness, as demonstrated by ThT and live cell FDAP imaging, possibly because of their increased pharmacokinetic properties driven by the presence of the fluorine atoms.In summary, the findings presented in this thesis provide a proof of concept that pre-structured hairpin mimics serve as a solid foundation for the development of innovative inhibitors of Tau and Aβ aggregation. The chaperone mimicry strategy has demonstrated its remarkable efficacy being the most potent dual inhibitor and, providing at the same time a new probe to simplify and study Tau-Hsp90 physiological interactions. The inclusion of the β2,2-isoxazoline amino acid into the peptide sequence allowed to demonstrate that the conversion of Tau toward an extended conformation, with the consequent exposure of PHF6* and PHF6, is the triggering event leading to Tau self-assembly.
USING B-STRAND AND B-HAIRPIN PEPTIDOMIMETICS INDUCED BY CONSTRAINED AMINO ACIDS TO MODULATE THE AGGREGATION OF THE MICROTUBULE ASSOCIATED PROTEIN TAU: DESIGN, SYNTHESIS AND EVALUATION / D. Di Lorenzo ; supervisor: S. Ongeri, M. L. Gelmi ; co-supervisor: N. Tonali. Università degli Studi di Milano, 2023 Jul 04. 35. ciclo, Anno Accademico 2022.
STUDY OF GRAPEVINE ROOTSTOCK RESPONSE TO WATER STRESS
More than 80% of vineyards around the world use grafted plants: a scion of Vitis vinifera grafted onto a rootstock of single or interspecific hybrids of American Vitis species, resistant or partially resistant to Phylloxera (Daktulosphaira vitifoliae (Fitch, 1856)). The genetic variability of grapevine rootstocks plays a fundamental role in their adaptation to the environment (Serra et al., 2013). In the climate change scenario, predicting an increase of aridity in the near future (Dai, 2013), the more frequent and severe drought events may represent the major constrain for the future of viticulture (IPCC, 2018; Schultz, 2000). Therefore, the selection of new rootstocks able to cope with unfavourable environmental condition is a key asset, as well as a strategy to improve crop yield/vegetative growth balance on scion behaviour (Corso and Bonghi, 2014). So far, the influence of rootstock on scion physiological performance during water stress has always aroused great interest. On the contrary, the scion impact on rootstock response is still less debated. Therefore, the effect of grafting on rootstock behaviour have been investigated. Phenotypical and large-scale whole transcriptome analyses on two genotypes, a drought-susceptible (101-14) and a drought-tolerant (1103 P), own-rooted and grafted with Cabernet Sauvignon, subjected to a gradual water shortage in semi-controlled environmental conditions have been performed. The ungrafted condition affected photosynthesis and transpiration, meaning the decisive role of scion in modulation of gas exchanges and in general in plant adaptation. Molecular evidence highlighted that the scion delays the stimulus perception and rootstock reactivity to drought.Since 1985, the DiSAA research group operating at the University of Milan is carrying on a rootstock crossbreeding program which has led to the release of four genotypes: M1, M2, M3 and M4. They show from moderate to high tolerance to drought (M4 > M1 = M3 > M2). In order to characterize their performance during water stress, their physiological (gas exchanges and stem water potential) and transcriptome response (genes involved in ABA-synthesis and ABA-mediated responses to drought) under well-watered and water stress conditions were examined. The behaviour of M-rootstocks (M1, M2 and M3) was compared with that of other commercial genotypes largely used in viticulture, either tolerant (140 Ru, 41 B, 110 R, 1103 P), less tolerant (SO 4, K 5BB) and susceptible (420 A and Schwarzman). Discriminant analysis (DA) showed that when water availability starts to decrease, rootstocks firstly perceives the stress activating a transcriptome response, consequently physiological changes have been observed. It also demonstrated that the three M-rootstocks were clearly discriminated: M4 was grouped with the most tolerant genotypes while M3 with the less tolerant or susceptible ones from a physiological standpoint, confirming their different attitude to tolerate water stress.M4 has proven to be a promising rootstock due to its ability to adapt to drought conditions. Considering the constant great demand for vine planting materials, the obtainment of genetically homogeneous populations (i.e. clones) from elite individuals through micropropagation represents a rapid alternative to conventional multiplication. For this reason, an efficient high-throughput protocol for M4 in vitro propagation was set up. Its attitude to shooting, root development and callus proliferation was compared to that of other rootstocks largely used in viticulture (K5BB, 1103P, 101-14 and 3309C). Moreover, pro-embryogenic and embryogenic callus from bud explants were also produced, representing a cellular material manipulable with the genetic engineering techniques. In water scarcity condition, among the mechanisms activated by M4, the great ability to scavenge ROS, related to the increased accumulation of stilbenes and flavonoids, may be such as to give it tolerance to
STUDY OF GRAPEVINE ROOTSTOCK RESPONSE TO WATER STRESS / L. Caramanico ; tutor: O. Failla ; co-tutor: G. De Lorenzis ; doctoral coordinator: D. Bassi. DIPARTIMENTO DI SCIENZE AGRARIE E AMBIENTALI - PRODUZIONE, TERRITORIO, AGROENERGIA, 2020 Feb 04. 32. ciclo, Anno Accademico 2019. [10.13130/caramanico-leila_phd2020-02-04].
ONSET OF ANEMIA IN PULMONARY DISEASES: ROLE OF HYPOXIA, OXIDATIVE STRESS, IRON METABOLISM AND HEMATOPOIETIC REGULATORS
BACKGROUND: In patients affected by severe hypoxia, such as those with Acute Respiratory Distress Syndrome (ARDS), an acute condition characterized by sudden onset of severe hypoxemia and a severe manifestation of COVID19, anemia is a common complication. It remains to be established weather iron metabolism, especially hepcidin, and oxidative stress are involved in negatively interfering with physiological hematopoietic compensatory mechanisms to hypoxia.AIMS:- To investigate etiology and progression of anemia in ARDS patients, through biomarkers of inflammation, iron metabolism and oxidative stress.- To identify the effects of increased hepcidin and the potentially protective effects of erythropoietin (EPO) in the energetic metabolism of lung and brain cells.METHODS: We collected blood samples at three timepoints from 20 ARDS patients: at the diagnosis (T0), when hemoglobin (Hb) was 3 g/dl less than T0 or <8 g/dL (T1), and two days after T1 (T2). Twenty ambulatory chronic obstructive pulmonary disease (COPD) patients were recruited to compare with non-anemic chronic hypoxia and ten non-hypoxic volunteers represented the control group. We reproduced hypoxia and inflammation in hypoxic lung A549 cells treated with lipopolysaccharide (LPS), and mitochondrial damage in neural SHSY5Y cells. The effect of hepcidin treatment and EPO on energetic metabolism was evaluated by measuring oxygen consumption and extracellular acidification rate (Seahorse technology).RESULTS: In ARDS patients, [Hb] decreased within 7-10 days after T0. We observed that ARDS induced higher IL6, hepcidin and malondialdehyde (MDA) with respect to both COPD and controls. The antioxidant barrier (ferric reducing antioxidant power test, FRAP) was weakened from T0 to T1. COVID19 ARDS values were similar to non-COVID ARDS, except for higher hepcidin and non-transferrin-bound iron (NTBI). EPO values were unchanged despite hypoxia. In hypoxic A549 cells, LPS decreased viability and, in combination with hepcidin, decreased mitochondrial respiration and increased the glycolytic rate. In SHSY5Y challenged cells, EPO partially rescued mitochondrial damage and increased glycolysis.CONCLUSIONS: ARDS is characterized by systemic release of inflammatory mediators, which negatively interfere with the hematopoietic adaptation to hypoxia. Oxidative stress and inflammation increase hepcidin, which induces a dysregulation in iron metabolism, with anemia and mitochondrial damage.
ONSET OF ANEMIA IN PULMONARY DISEASES: ROLE OF HYPOXIA, OXIDATIVE STRESS, IRON METABOLISM AND HEMATOPOIETIC REGULATORS / S. Ottolenghi ; coordinatore: C. Sforza ; tutor: M. Samaja. Università degli Studi di Milano, 2021 Mar 17. 34. ciclo, Anno Accademico 2020. [10.13130/ottolenghi-sara_phd2021-03-17].
EXPLORING THE PRE-LEUKEMIC PHASE OF ACUTE PROMYELOCYTIC LEUKEMIA: INSIGHTS FROM STUDIES OF SINGLE CELL OMICS
Acute Promyelocytic Leukemia (APL) is a distinct subtype of Acute Myeloid Leukemia (AML) driven by the oncogenic fusion protein PML-RAR, which acts as an aberrant transcription factor and blocks myeloid maturation at the promyelocyte stage. In vivo, the expression of PML-RAR is necessary but not sufficient for full-blown leukemia: knock-in (KI) mouse models of APL undergo a prolonged pre-leukemic phase during which hematopoiesis appears normal. This stage provides a unique opportunity to investigate the earliest molecular and cellular alterations that precede the disease.To dissect early events, a transgenic PML-RAR model was used in combination with high-resolution single-cell technologies. Hematopoietic stem cells (HSCs), common myeloid progenitors (CMPs), and a residual and more differentiated lineage-negative Sca-1-c-Kit- (LSK-) subpopulation were profiled via bulk RNA-seq, single-cell RNA-seq (scRNA-Seq), and single-cell ATAC-seq (scATAC-Seq). The integrated analysis revealed that PML-RAR expression maintains largely unaltered the overall transcriptional program of pre-leukemic cells, while driving cell cycle and a selective expansion of specific cell subtypes, correlated with a reduction of chromatin accessibility at several genomic loci.Functional assays corroborated these findings. Transplantation experiments demonstrated that PML-RAR-expressing cells from the three hematopoietic compartments gave rise to transplantable leukemia, albeit with different efficiency. Of note, following lethal irradiation, these cells provide radioprotection and long-term repopulation in recipient mice, indicating that PML-RAR confers an advantage that becomes particularly evident in perturbed conditions.Comparison with published datasets further reinforced our observations. A transcriptional signature directly regulated by PML-RAR expression identified in human NB4 APL cells was detected in our analysis of single-cell data. Notably, the signature was confined to the expanded subpopulations, despite differences in species and experimental context. This supports the idea that expanded pre-leukemic progenitors represent the origin of transformation.Together, these results indicate that PML-RAR is able to initiate leukemogenesis through a dual program of chromatin repression and cell cycle dysregulation, selectively expanding progenitor subsets with leukemogenic potential. By integrating transcriptomic, epigenomic and functional data, this work provides a comprehensive map of the earliest oncogenic events of APL, highlighting candidate cell populations from which leukemia may rise. Also, these findings advance the understanding of pre-leukemic biology and suggest novel therapeutic strategies which may intercept disease at its earliest stages.
EXPLORING THE PRE-LEUKEMIC PHASE OF ACUTE PROMYELOCYTIC LEUKEMIA: INSIGHTS FROM STUDIES OF SINGLE CELL OMICS / E. Villa ; tutor: S. Minucci ; phd coordinator: D. Pasini. Dipartimento di Oncologia ed Emato-Oncologia, 2025 Oct. 37. ciclo, Anno Accademico 2024/2025.
CLINICAL APPLICATIONS OF PARAOXONASE-1 IN DOMESTIC ANIMALS: BEYOND CONVENTIONAL INFLAMMATORY MARKERS
Inflammation and oxidative stress frequently occur in domestic animals. The diagnosis and monitoring of inflammatory and oxidative conditions are based on markers that are so far considered accurate but may lack specificity and prognostic utility. In the last years’ research, interest has grown about the role of Paraoxonase-1 (PON-1) in the pathogenesis and diagnosis of many diseases characterized by inflammation and/or oxidative stress, both in human and veterinary medicine. In veterinary species, PON-1 measurement is to date primarily restricted to the research field and has not been yet undertaken in the routine clinical field. However, recent research has highlighted promising utility in many clinical contexts. Since accurate and precise spectrophotometrical assays validated in many species are now available and proved to be cheap, rapid and automated, its measurement could easily be included in routine chemistry panel in the near future. Therefore, the aim of my research was focused on the evaluation of the usefulness of PON-1 as a diagnostic and prognostic marker in different conditions characterized by inflammation and/or oxidative stress in different veterinary species.The results collected in the different studies described in this thesis provided further insights about the possible utility of PON-1 as a biomarker in veterinary clinical pathology. We confirmed the role of PON-1 as a marker of systemic inflammatory response syndrome, possibly associated with sepsis in horses and dogs, and its usefulness as a prognostic marker in these contexts. Furthermore, in dogs we confirmed its usefulness as a marker of disease severity and its possible applications as a tool for clinical decision or detection of subclinical disease. As regards cats and pigs, results about its usefulness are still preliminary and deserve further investigation with a wider and more standardized caseload. On the contrary, we demonstrated the usefulness of PON-1 as an ancillary test for clinical decisions or to address further testing in cattle with bovine respiratory disease.
CLINICAL APPLICATIONS OF PARAOXONASE-1 IN DOMESTIC ANIMALS: BEYOND CONVENTIONAL INFLAMMATORY MARKERS / D. Scavone ; tutor: S. Paltrinieri ; coordinator: F. Ceciliani. Università degli Studi di Milano, 2022 Sep 06. 34. ciclo, Anno Accademico 2021.
NARRARE LA MORTE.STORIA POLITICA DEL LUTTO A ROMA DALLA REPUBBLICA AL PRINCIPATO AUGUSTEO.
Within a diachronic and synchronic analysis, this work explores the metamorphosis that the relationship between power and the funerary sphere underwent in Rome over time, from the early days of the Republic to the death of Augustus. The social context in which the funerary operations came to light, as well as their utilization in the service of political competition, areexamined both in terms of performative outcomes (with particular reference to public or proto-public funerals) and literary outcomes (with particular reference to laudationes funebres and consolationes).Chronologically, the timeframe under consideration is divided into three periods, each presenting affinities that allow the definition of homogeneous modules and recurring trends. An introductory section is dedicated to the Early and Middle Republican Era and aims to provide a foundation for understanding the origins of phenomena that will emerge more fully from the 1st century BCE. Concerning the Late Republican Era, the analysis beginswith the funeral of Sulla, conventionally considered the first funus publicum celebrated in Rome. Caesar plays a significant role in this section, both for the dynastic ambitions that he early incorporated into his funerary programs, and for the funeral held in his honor, the first of a series of seditiosa funera that increasingly occupied the rostra. Cicero deserves special mention in this phase, being the author of elaborate plans around the spectacularization ofthe ‘deaths of Modena’, as well as a vast literary production on the theme (Consolatio ad se ipsum and Laus Catonis, to which Caesar responded with the Anticato). These writings, though fragmentary, are examined to highlight both the (typically Ciceronian) dialectic between private-therapeutic and public-political function, and the choice of genre itself, which appears to be ‘a safe form to use for a political statement’ in the temporal upheaval of theyears 46-45 BCE. The third and final section of the analysis focuses on the Augustan Era and aims to clarify the ways in which the Princeps gave rise not only to a monopoly of performative practices at the rostra, but also to a general resemanticization of occasions and places of traditional convergence, defining a ‘funerary protocol’ that, for the first time, transcended the boundaries of the Urbs and impacted the spaces and times of the empire as a whole. The analysis of Consolatio ad Liviam and Elegiae in Maecenatem, on the other hand, completes the outlined picture, showing the different attitudes with which different social, cultural, and generational groups experienced and interpreted the assertion and subsequent transmission of a new power.
NARRARE LA MORTE.STORIA POLITICA DEL LUTTO A ROMA DALLA REPUBBLICA AL PRINCIPATO AUGUSTEO / L. Fontana ; tutor: S. Segenni ; coordinatore del dottorato: F. Slavazzi. Università degli Studi di Milano, 2024 Jun 19. 36. ciclo
PTH RESISTANCE SYNDROMES: AGE-DEPENDENT CLINICAL PICTURE AND EMERGING UNCOMMON FEATURES
BACKGROUND: The term “PTH resistance syndromes” refers to a group of diseases so far named pseudohypoparathyroidism (PHP). The improvement of knowledge on pathophysiology and genetic background led to the proposal of a new classification that encompasses all the diseases caused by mutations within the PTH/PTHrP signaling cascade (iPPSD). PTH resistance is the hallmark of these disorders, but clinical picture is wider and more complex, with high variability and age-dependent presentation. A diagnostic delay is commonly reported; thus, our studies aim to describe some emerging uncommon clinical features with the final goal to improve diagnostic and management of these syndromes.METHODS: We conducted three retrospective analysis collecting data from large cohorts of PHP/iPPSD patients and exploring some still undescribed aspects (neonatal complications, early infancy features, hypercalcitoninemia, skeletal phenotype). As for Study A and Study B, we have collaborated with a referral Paediatric Center in France, therefore we could describe also paediatric features and rely on a very large cohort of patients. Study A focused on the rate of neonatal complications occurring in each PHP/iPPSD category during the first month of life, trying to estimate potential predictors and to correlate neonatal complications with the severity of the disease. Moreover, in Study A, we extracted from each medical record data about the age of onset or the age of diagnosis of the main clinical and biochemical features, aiming to describe early natural history of the disease and to explore the need of age-dependent criteria for diagnosis. The main aim of Study B was to investigate the prevalence and the dynamics of hypercalcitoninemia in PHP/iPPSD patients, and to define the clinical impact of this hormonal feature, also by analyzing data about thyroid ultrasound (US) and calcium stimulation test performed in a subset of patients. Study C is still ongoing, but we present preliminary data and an interesting case report. This study attempts to define the extent of bone responsiveness to elevated PTH levels, through the analysis of bone turnover markers, bone mineral density and fractures’ prevalence. Most of our patients had received a molecular diagnosis. RESULTS: • Study A: the first cohort was made up of 136 patients with a diagnosis of PHP/iPPSD. At least one neonatal complication occurred in 36% of patients and in 38.7% of cases neonatal complications were considered as severe, because they required prolonged hospitalization or intensive care assistance. PHP1A/iPPSD2 patients represent the majority of our cohort and 47% of them experienced neonatal complications, being the category the most affected (p=0.002). Neonatal hypoglycemia and transient respiratory distress appeared significantly frequent in this latter group, i.e., 10.5% and 18.4%, respectively. Neonatal complications were associated with the risk of developing neurocognitive impairment later in life (p=0.02) and earlier TSH resistance (p<0.001). In the second cohort (n=117) the median age at diagnosis was 5.9 years old. Age of onset of PTH resistance and brachydactyly, major criteria for diagnosis, was significantly different from that of both TSH resistance and obesity (median age 6.1, 5.8, 1.85 and 2 years, respectively). Minor diagnostic criteria were more represented than major criteria in children before 2 years (p=0.002). Indeed, in 64% of patients before 2 years none of the major criteria was described, conversely 71% had already developed at least 1 minor criteria; in particular, 20% had developed TSH resistance and obesity.• Study B: in the paediatric cohort 65.9% had hypercalcitoninemia (median CT 15 ng/L); in the adult cohort 53.5% (mean CT 21.6 ng/L). There was no difference between CT in paediatric and adult population; we observed stable CT levels over a median follow-up of 134.5 months in adults. Notably, no correla
PTH RESISTANCE SYNDROMES: AGE-DEPENDENT CLINICAL PICTURE AND EMERGING UNCOMMON FEATURES / G. Del Sindaco ; coordinatore: M. Del Fabbro ; tutor: G. Mantovani. Università degli Studi di Milano, 2024 Dec 19. 37. ciclo, Anno Accademico 2023/2024.
RETURN TO SPORT AFTER ACHILLES TENDON RUPTURE
Background: Achilles tendon (AT) rupture is a common injury, particularly among physically active individuals, with a 30% growth rate in the most affected age group (30–39 years). Among professional athletes, 25% fail to return to sport (RTS), and those who do typically require 8–11 months of recovery. Furthermore, their performance can decline by up to 50% during the two years following RTS, and their career length is shorter compared to non-injured players or those with anterior cruciate ligament (ACL) injuries. Long-term deficits after AT repair have been documented up to 10 years post-injury, including tendon elongation, reduced calf circumference, atrophy of type-1 muscle fibers, decreased heel-raise test performance, and reduced ability to generate force in a plantar-flexed position. Additionally, low psychological readiness to RTS, or high levels of kinesiophobia at six months may influence RTS outcomes. Given these suboptimal RTS outcomes and the lack of consensus on optimal treatment strategies for AT rupture, there is a need to explore potential areas for improvement in the RTS process. Aims: These considerations have inspired my PhD research project, which aims to identify gaps in the current AT RTS process and provide insights to optimize outcomes for patients recovering from AT rupture. The specific aims of the three research projects are as follows: 1. To map the literature regarding which criteria are used to clear athletes for RTS after AT repair. Understanding the criteria defined in the literature allows for clarity on the standards required for RTS and the identification of potential gaps area in the RTS process that lack clear guidelines. 2. To observe and analyze the impact of AT rupture in ankle movement quality during the mid-stage of rehabilitation. The comparison between the repaired and unrepaired limbs aims to identify potential differences in movement quality between the two ankles. 3. To describe the on-field rehabilitation (OFR) workload of football players returning to sport after AT repair. Global Positioning System (GPS) monitoring workloads during the late-stage of rehabilitation may be a key factor to lead load progression during the late-stage of rehabilitation, providing insights to prepare football players for the demands of the return to training (RTT).Methods: 1) Project one involved a Scoping Review conducted in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping Reviews (PRISMA-ScR) Checklist. A combination of search terms and Boolean operators, regarding AT rupture and RTS criteria, was used to search the literature from 1995 to 2022 across three databases: Web of Science, Scopus, and MEDLINE. Papers inclusion and exclusion criteria were defined. A data charting was create to collect criteria, and the insights regarding AT rehabilitation outcome. 2) Project two was a case series involving 23 sportive patients who underwent AT repair. The inclusion criteria for participants were: being treated at one of the Isokinetic Medical Group Clinics, having undergone Achilles tendon (AT) repair surgery, and performing a bilateral depth jump. To observe the ankle movement quality the ankle joint angle (γ angle), and peak vertical ground reaction force (peak VGFR), in both repaired and unrepaired limbs during the eccentric and concentric phase of a bilateral depth jump where assessed. To test for differences between the repaired and unrepaired sides, variables were analyzed collectively using a multivariate Mixed Linear Model with repeated measures. Effect sizes and confidence intervals were also computed. 3) Project three was a pilot study with a small cohort including 5 male football players who underwent AT repair and were monitored during the OFR. Workload data for each OFR session were collected using GPS and heart rate tracking. Variables measured and reported were: PL, Player load (A
RETURN TO SPORT AFTER ACHILLES TENDON RUPTURE / M.f. Busa' ; supervisor: F. Esposito ; co-supervisor: M. Zago ; executive supervisor: F. Della Villa ; chairs of the program: F. Esposito, C. Gozzoli. Dipartimento di Scienze Biomediche per la Salute, 2025 Mar 26. 37. ciclo, Anno Accademico 2023/2024.
ORIGEN Y ANALISIS DEL TURISMO ESCOLAR: EL TOUR DIDACTICO COMO PRODUCTO TURISTICO Y ACTIVIDAD COMPLEMENTARIA PARA POTENCIAR LAS COMPETENCIAS CLAVE DEL ALUMNADO
Pre-university education offers students a unique opportunity for tourism through the complementary and extracurricular activities programmed by the departments of primary and secondary schools, initiatives previously approved by the school council and which teachers use as a didactic resource to motivate their students in their subjects. These school trips become real tourist experiences by offering pupils a fun and educational moment with a well-defined product, supported by experts in the sector who can benefit economically by providing services adapted to the nature of the group and even directly, in collaboration with the teacher, by coordinating, accompanying and interpreting the different tourist attractions in the areas close to the school premises.From a methodological point of view, the research has been approached through an empirical approach using mixed descriptive techniques with the intention of justifying a tourist typology focused on school trips. By creating a travel agency specialised in educational guided tours, favouring the dynamic use of the participant-observation methodology, resulting in a tourist-educational product validated and tested by the schools.The purpose of this research is: (1) to unravel school tourism as a sub-segment of educational and cultural tourism from its origins to the present day. To this end, the market generated is analysed, well defined in a network of trips and products designed by tourism professionals, materialised in school outings, excursions, trips, cultural exchanges, workshops and activities. (2) To promote initiatives outside the classroom as a didactic strategy and socio-economic activity that creates synergies between the school community and tourism service providers. (3) To create an educational tour validated by teachers and applied to groups of schoolchildren as a complementary, transversal activity that helps to strengthen the key competences decreed by the European Commission and implemented in the education systems of the member countries, as in the case of Spain and Italy. (4) Include in the last part of the work the business plan of a tourism company, set up during the research process, to serve as an example for those professionals in the sector who want to revitalise this type of tourism-educational activities aimed at schools to discover their territory with the help of a heritage interpreter specialised in school groups.
ORIGEN Y ANALISIS DEL TURISMO ESCOLAR: EL TOUR DIDACTICO COMO PRODUCTO TURISTICO Y ACTIVIDAD COMPLEMENTARIA PARA POTENCIAR LAS COMPETENCIAS CLAVE DEL ALUMNADO / F.m. Romero Sanchez ; directores: E. Asenjo Rubio, D. Gavinelli. Università degli Studi di Milano, 2024. 30. ciclo
FURTHER INSIGHT INTO V-ATPASE ROLE IN GLIOMA STEM CELLS
V-ATPase is a proton pump mainly localized on lysosomes and on plasma membrane of specialized cells. It is responsible of proton translocation and acidification of intra- and extra-cellular environment. Our group demonstrated that the over-expression of the subunit G1 (V1G1) is involved in the maintenance of the stem cell niche in glioblastoma (GBM) and correlates with poor prognosis in GBM patients. In this work we aimed to elucidate the role of V-ATPase in GBM stem cells from a functional perspective. We demonstrated that neurospheres (NS) with higher levels of V1G1 subunit (High-V1G1), compared with NS with lower levels of V1G1 (Low-V1G1), were characterized by increased clonogenicity in vitro and in vivo, invasiveness, lysosomal acidification and ERK pathway activation. Specific inhibition of V-ATPase activity, by Bafilomycin (BafA1), but not of ERK or other lysosomal drugs, induced reactive oxygen species (ROS)-mediated apoptosis only in High-V1G1 NS. In addition, BafA1 treatment affected mitochondria homeostasis only in High-V1G1 NS. Preliminary experiments suggested that a V-ATPase pump might be localized on the mitochondria or it could mediate direct contacts between mitochondria and lysosomes thus causing an imbalance of charges (proton flux) when perturbed. Finally, High-V1G1 and Low-V1G1 NS differed in terms of metabolic behaviours: preferential use of glycolysis by Low-V1G1 NS opposite to use of oxidative metabolism in High-V1G1 NS. V-ATPase block by BafA1 in High-V1G1 NS shifted their metabolism to that of Low-V1G1 NS. On the other hand, the autophagic pathway, that is directly connected with lysosomal function, was blocked by BafA1 only in Low-V1G1 NS. These phenotypes were not modulated by ERK or lysosomal acidification inhibitors alone, indicating a specific role for V-ATPase proton pump in modulating them. Taken together, these results indicate that V-ATPase is crucial for GBM stem cells viability through different mechanisms that include bioenergetics sensing and requiring, mitochondrial homeostasis and ERK signalling activity.
FURTHER INSIGHT INTO V-ATPASE ROLE IN GLIOMA STEM CELLS / A.m. Storaci ; tutore: S. Ferrero ; co-tutore: V. Vaira ; coordinatore: M. Samaja. Università degli Studi di Milano, 2020 Jan 17. 32. ciclo, Anno Accademico 2019. [10.13130/storaci-alessandra-maria_phd2020-01-17].
CHARACTERIZATION OF NON-EQUILIBRIUMFLUCTUATIONS DURING FREE-DIFFUSION INHIGHLY-STRATIFIED SOLUTIONS OF GLYCEROL ANDWATER
We present two topics, both in the area of transport mechanisms(mass diffusion or heat transport) occurring in fluids. The first andmain topic is a study of the non equilibrium fluctuations (NEF) thatoccur during isothermal free diffusion between two solutions ofglycerol and water with various concentration differences. Thenonlinearity of the system, determined by the strong stratification ofthe sample, requires the introduction of an interpretive modelcapable of characterizing the dependence of the correlationproperties of the non-equilibrium fluctuations on the localthermophysical variables of the system. The proposed model makesit possible to characterize the dynamics of non-equilibriumfluctuations in the presence of a wide range of relaxation timesdetermined by the strong stratification of the sample, unlike thecumulative methods commonly used in dynamic light scatteringexperiments, which work well in the presence of moderate dispersionof relaxation times. The second topic concerns the convectivestructures that appear in a thermophilic suspension of nanoparticlesheated from below. Such a system presents a complex stabilitydiagram, determined by the competition between the stabilizing flowof nanoparticles induced by thermophoresis and the destabilizingflow determined by thermal convection. We analyzed Rayleigh-Bénard convection in a suspension of highly thermophilicnanoparticles with a large negative separation ratio Ψ = −3.5 heatedfrom below. We demonstrated the presence of transient stateslocalized in the range of Rayleigh numbers 2200 < Ra < 3000. Thesestates rotate rapidly around their axis and gradually decrease in sizeuntil the system returns to a purely conductive state. This secondtopic is not totally separate from the first one, since the study of thestability diagram is the first step toward studying the thermal andconcentration fluctuations generated by heating a thermophiliccolloidal suspension from below.
CHARACTERIZATION OF NON-EQUILIBRIUMFLUCTUATIONS DURING FREE-DIFFUSION INHIGHLY-STRATIFIED SOLUTIONS OF GLYCEROL ANDWATER / S. Castellini ; supervisore: A. Vailati ; tutor: Marina Carpineti ; coordinator: M. Paris. Dipartimento di Fisica Aldo Pontremoli, 2023 Jul 25. 35. ciclo, Anno Accademico 2022.
UNVEIL THE CONNECTION BETWEEN METABOLIC AND INFLAMMATORY CARDIOVASCULAR RISK FOSTERED BY NUTRIENT INTAKES.
Background: Therapeutic strategies targeting the residual inflammatory risk (increased levels of inflammatory markers) effectively further reduced ACVD incidence. On the other hand, therapeutic strategies to contain the residual metabolic risk (increased TGRL) provided discordant results. Recent evidence support a casual association between metabolic and inflammatory risk, and also describe, at a molecular level, the immuno-metabolic effects of dietary intakes, and the potential implication of gut microbiome in these connections. In this context, we aimed to unveil the connection between the metabolic and inflammatory cardiovascular risk fostered by nutrient intakes. To achieve our aim the following tasks were settled: (i) deepen the relation between the metabolic and inflammatory risk, beyond LDL-C; (ii) identify the targetable dietary aspects to reduce both the metabolic and inflammatory risk; (iii) constraining the metabolic and its associated inflammatory risk by long-term targeting of those dietary aspects; (iv) identify the potential implications of gut microbiome composition in the association between dietary intakes, and the metabolic and inflammatory ACVD risk. Materials and methods: In two cohorts, one in primary and the other in secondary prevention for ACVD (PLIC-Progressione delle Lesioni Intimali Carotidee Study, SMART- Secondary Artery Manifestation Study), were compared the association of either the metabolic (as calculated TGRL-C levels) risk or LDL-C levels with the inflammatory risk, taking advantage of a wide set of proteins measured in plasma (Olink platformTM) and predicting their involvement in immune-inflammatory processes through Ingenuity Pathway Analysis. In the PLIC cohort, the same set of plasma proteins was employed to validate the predicted inflammatory potential of diet (estimated from the analysis of 7-day dietary record via the Dietary inflammatory Index, DII). Then, we explored the relation of the DII with circulating TGRL concentrations and triglycerides/cholesterol content of TGLR in serum samples analyzed by Nuclear Magnetic Resonance (Nightingale Health platform). Seven-teen subjects, from the PLIC cohort, were enrolled in a 4-week dietary interventional trial to test the effects of lowering the DII on the long-term metabolic and inflammatory risk but also on the acute metabolic and inflammatory risk fostered by the consumption of a pro-inflammatory/fat-enriched meal (OFL). Long-term and acute metabolic and inflammatory risk were measured as i) fasting, ii) postprandial, and iii) postprandial increase in TGRL triglycerides and cholesterol content and immune cells count and their markers expression levels. With an ex-vivo setting, we explored the acute and long-term effects of fasting and postprandial TGRL on inflammatory gene expression in immune cells before and after lowering the DII. We finally observationally correlated gut microbiome composition (assessed through Next-generation Sequencing analysis on DNA extracted from fecal samples) with the DII, and TGRL features from the Nightingale Health platform. Results: We found, both in primary and secondary prevention settings, a higher number of circulating plasma proteins associated with increased TGRL-C compared to increased LDL-C. On top of common ACVD risk factors (CVRFs) the performance of classifications of these proteins was impressively higher in identifying those subjects with increased TGRL-C compared to those with increased LDL-C. Furthermore, these proteins recapitulated the involvement of immune cell inflammatory processes. The inflammatory potential of diet (validated against circulating plasma proteins and inflammatory processes) marked increased levels of circulating TGRL and their triglycerides and cholesterol content. The dietary intervention resulted in a significant reduction in the inflammatory potential of diet and effectively improved both the long-term and acute metabolic and inflammatory risk. Some gut mic
UNVEIL THE CONNECTION BETWEEN METABOLIC AND INFLAMMATORY CARDIOVASCULAR RISK FOSTERED BY NUTRIENT INTAKES / E. Mattavelli ; tutor: A. Baragetti, P. Magni; coordinatore: G.D. Norata. Università degli Studi di Milano, 2024 Apr 18. 36. ciclo, Anno Accademico 2023.
DENKMALER UND SACRARI FUR DIE HELDEN DES FASCHISMUS IN ITALIEN 1922 - 1943
Questo elaborato di tesi intitolato Denkmaeler und sacrcari fuer die Helden des Faschismus in Italien 1922-1943 si prefigge di indagare le opere architettoniche e scultoree ideate o realizzate su iniziativa del partito fascista durante il ventennio 1922-1943, al fine di commemorare i caduti per la causa del regime, presentando le strategie adottate in ambito architettonico e scultoreo per ottemperare a direttive ed esigenze del culto del littorio. Innanzitutto è necessario premettere che si tratta di un lavoro di ricerca scientifico, che prende le distanze da qualsiasi posizione politica, basato sulla ferma convinzione che sia doveroso occuparsi della storia e comprenderla appieno, con il fine ultimo di impegnarsi a tutelare la democrazia.Per il movimento fascista il culto dei caduti fu “probabilmente il più espressivo del suo senso di religiosità secolare” (Gentile, 1993, p. 47) e svolse fin dai primordi un ruolo chiave nella sua ideologia, costituendo una fra le manifestazioni più durature e meno soggette a variazioni, caratterizzato da una dimensione locale e al contempo capillarmente presente su tutto il territorio nazionale. Attingendo, come noto, alle forme e ai linguaggi del primo dopoguerra, così come al repertorio cristiano, il regime dedicò monumenti e sacrari ai caduti per la causa del regime (Suzzi Valli 2008; Masseroni, 2017) al fine di legittimare il proprio culto. Appuratone il carattere sincretico, i riferimenti alla religione cristiana, al contesto militare del secolo precedente ed alla romanità, questa tesi si propone di analizzare le architetture, il repertorio iconografico e il rapporto tra modernità e classicità adottati dal regime per definire architettonicamente e figurativamente la sacralità dei suoi luoghi sacri. Con questo elaborato si offre quindi un contributo alla riflessione relativa alla definizione del concetto di sacrale nell’architettura e nell’arte del culto del littorio, suggerendo strumenti e chiavi di lettura di questo fenomeno, a partire dall’analisi delle fonti primarie – edite ed archivistiche – risalenti al ventennio. Considerati il carattere capillare di diffusione dei monumenti – che rimanda a quello della monumentomania del primo dopoguerra – e il numero di progetti e realizzazioni all’interno delle case del fascio, stimato a circa 25.000 (Mangione, 2006), risulta evidente come numerosi siano i monumenti e sacrari per i caduti per la causa fascista ad oggi non compiutamente approfonditi dalla ricerca. Nell’indagare quest’ambito, si palesa l’ulteriore difficoltà della definizione terminologica di sacrario e della tipologia in cui questi si inquadrano. Definito il sacrario come “Cappella o ambiente in cui sono raccolti i resti o i ricordi di persone benemerite della patria” (Treccani, enciclopedia), i luoghi sacri dedicati ai caduti fascisti si differenziano dai monumentali progetti di sacrari militari in quanto i primi si caratterizzano per un contesto di realizzazione più ampio, che spazia da ambienti ricavati all’interno di edifici istituzionali – fra cui appunto le case del fascio –, inseriti in punti nevralgici del tessuto urbano, a quelli previsti all’interno di edifici religiosi, in cimiteri così come include quelli allestiti in contesti espositivi. Ad oggi la ricerca riconduce, correttamente, i numerosi progetti di sacrari alle forme e ai linguaggi del noto modello del sacrario liberiano della Mostra della Rivoluzione Fascista. Tale tesi viene supportata dalla dimensione prescrittiva della realizzazione di un ambiente dedicato ai caduti all’interno delle case del fascio, contenuta nel foglio d’ordini promulgato dal partito il 20 novembre 1932. Si ritiene tuttavia che – considerando l’eterogeneità dei differenti contesti di realizzazione, la varietà di forme, stili e linguaggi impiegati nei vari progetti – la tipologia di monumenti e sacrari nel suo complesso vada ulteriormen
DENKMALER UND SACRARI FUR DIE HELDEN DES FASCHISMUS IN ITALIEN 1922 - 1943 / M. Tomesani ; tutor: I. Herklotz, S. G. Bignami ; coordinatore: F. Slavazzi. Dipartimento di Beni Culturali e Ambientali, 2024 Mar 14. 35. ciclo [10.13130/tomesani-marta_phd2024-03-14].
CHARACTERIZATION OF AIR POLLUTION AND AIR QUALITY USING INNOVATIVE SAMPLING METHODS AND ANALYTICAL TECHNIQUES FOR REGULATED AND NON-REGULATED POLLUTANTS
Air pollution is a critical global health and environmental issue, with major health risks associated with pollutants such as particulate matter (PM), ozone (O3), carbon monoxide (CO), and nitrogen oxides (NOx). Despite advancements in air quality regulations, many pollutants remain unregulated, both outdoors and indoors. Additionally, several sources of air pollution continue to be unmonitored, with their impacts on air quality yet to be fully understood. This thesis addresses two key research areas aimed at improving air quality monitoring and mitigation efforts. The first investigates the emissions from pizza ovens, a largely overlooked source of pollution in urban areas. Using advanced sampling methodologies and analytical techniques, this study assesses the contribution of these ovens to air quality. Emission factors for major pollutants (NOx, CO, OGC, TSP, PM10, PM2.5, PAHs) were determined through experimental testing, and their impact on urban air quality was evaluated through a case study in Milan. The second explores air quality monitoring in indoor sites, specifically schools and places of worship, using innovative sensors and monitoring techniques to evaluate a wide range of pollutants. The results underscore the importance of continuous monitoring to better understand the nature and impact of indoor pollutant sources, as well as to identify critical air quality issues in these spaces. The dual focus on both outdoor and indoor air quality aims to provide comprehensive insights into the evolving challenges of air pollution and the need for effective, targeted measures to protect public health and the environment.
CHARACTERIZATION OF AIR POLLUTION AND AIR QUALITY USING INNOVATIVE SAMPLING METHODS AND ANALYTICAL TECHNIQUES FOR REGULATED AND NON-REGULATED POLLUTANTS / A. Bergomi ; tutor: P. Fermo ; co-tutor: V. Comite, G. Migliavacca ; coordinatore: D. Passarella. - Università degli Studi di Milano. Dipartimento di Chimica, 2025 Mar 05. 37. ciclo, Anno Accademico 2023/2024.
INTERACTOMICS FOR RICE FLOWERING: A PROXIMITY LABELLING APPROACH
Flowering regulation in rice (Oryza sativa) involves a complex induction pathway that senses environmental inputs in the leaves and transmits signals to the shoot apical meristem (SAM) for reproductive differentiation. The transcription factor Hd1, homologous to Arabidopsis CONSTANS, is pivotal photoperiodic sensor, promoting flowering under short-day (SD) and repressing it under long-day (LD) conditions. Hd1 function is modulated through the interaction with other flowering regulators, such as Ghd7 and NF-Y proteins, to activate or repress the transcription of the florigen genes Hd3a and RFT1. A recently characterized florigen-like protein, called FTL1, contributes to meristem differentiation during inflorescence and panicle development, relying on interactions with OsFD transcription factors. During my PhD, I studied Hd1 and FTL1 function with traditional transcriptional and biochemical techniques, as well as through innovative proteomic approaches. Proximity Labelling (PL) was employed for the first time in our laboratory to explore novel protein-protein interaction of Hd1 and FTL1. PL relies on fusing TurboID (TbID), an optimized biotin ligase, to the protein of interest to obtain specific tagging of its interactors in vivo. Identification of the proximal proteome is enabled by selective precipitation of biotinylated proteins and mass spectrometry analysis. Our research has established an efficient PL protocol in agroinfiltrated N. benthamiana leaves and transgenic rice plants expressing TbID-fused proteins. TbID-fused FTL1 and Hd1 were successfully created, and protein analyses confirmed their stability and enzymatic efficiency, helping refining the optimal PL pipeline. A preliminary PL analysis was carried out on Hd1-TbID using agroinfiltrated N. benthamiana leaves. Furthermore, application of PL on rice FTL1-TbID floral meristems yielded >3000 proteins, with 102 identified as potential FTL1 interactors, revealing associations with chromatin, RNA metabolism, and cell structural elements (cell wall, plasmodesmata and microtubules). Ongoing research promises additional proteomic information, while validation experiments are underway to verify the novel information retrieved on FTL1 localization and function. With these experiments, we demonstrate that PL can expand our understanding of rice flowering regulating network, and that the obtained data can be a valuable resource to maximize the breeding of cultivated rice varieties.
INTERACTOMICS FOR RICE FLOWERING: A PROXIMITY LABELLING APPROACH / D. Chirivi ; tutor: F. Fornara ; coordinatore: R. Mantovani. - Milano. Dipartimento di Bioscienze, 2024 Jun 21. 36. ciclo
CRYSTAL CHEMISTRY AND PHYSICAL-CHEMICAL BEHAVIOR OF REE-BEARING PHOSPHATES AND ARSENATES: THE CASE STUDY OF MT. CERVANDONE
The crystal chemistry, crystal structure and behavior at non-ambient conditions of natural Rare Earth Elements-bearing phosphates [monazite-(Ce), nominally Ce(PO4), and xenotime-(Y), nominally Y(PO4)] and arsenates [gasparite-(Ce), nominally Ce(AsO4), and chernovite-(Y), nominally Y(AsO4)], from the hydrothermal quartz-bearing fissures outcropping at Mt. Cervandone (Lepontine Alps, Piedmont, Italy), have been investigated. The chemical and structural characterization, performed via electron microprobe analysis, Raman spectroscopy and single-crystal X-ray diffraction, showed that the zircon-type minerals chernovite-(Y) and xenotime-(Y) share a very similar (Y,HREE) composition and the same can be stated for the LREE content of the two monazite-type minerals [gasparite-(Ce) and monazite-(Ce)]. An almost complete solid solution has been found between xenotime-(Y) and chernovite-(Y), while a wide miscibility gap has been observed within the monazite series minerals of Mt. Cervandone. Moreover, both the unit-cell and the REE-coordination polyhedral volumes are strongly controlled by the cationic population at the T-site: an increase in As not only expands the volume of the TO4 tetrahedron, but even that of the REE-polyhedron, irrespective of the A-site population. An exception is provided by the relative abundance of Th and Ca at the A-site, which was found to expand the coordination polyhedron and unit-cell volumes irrespective of the T-site composition.The comparative analysis of the thermo-elastic behavior of selected samples has been conducted by in situ high-P, high-T and combined HP–HT X-ray diffraction experiments using both single-crystals and polycrystalline samples at synchrotron or conventional lab facilities. The compressional behavior of the investigated minerals is compatible with the literature data on the synthetic endmember counterparts. The two zircon-type minerals undergo different phase transitions under compression, while the monazite-type minerals experience a change in the compressional behavior, likely induced by the increase in coordination number of the REE-bearing polyhedron. The thermal expansion behavior of monazite-(Ce) and xenotime-(Y) confirm the literature data available, while the studied chernovite-(Y) is significantly less expandable than the synthetic counterparts described in the literature. The phosphates were found to be less compressible, but more expandable than the isostructural arsenates. The structural analysis at non-ambient conditions has been carried out on the basis of the refined structure models, allowing the description of the deformation mechanisms accommodating the bulk compression or expansion at the atomic scale. Both are mainly accommodated by the REEOx coordination polyhedra, while the tetrahedra behave as a quasi-rigid units. In conclusion, all the experimental data confirm the central role played by the T-site in controlling the structural deformation and, in turn, the bulk thermal expansion and compression.
CRYSTAL CHEMISTRY AND PHYSICAL-CHEMICAL BEHAVIOR OF REE-BEARING PHOSPHATES AND ARSENATES: THE CASE STUDY OF MT. CERVANDONE / F. Pagliaro ; tutor: P. Lotti ; cotutor: A. Guastoni ; coordinatore: M. I. Spalla. Dipartimento di Scienze della Terra Ardito Desio, 2023 Apr 17. 35. ciclo, Anno Accademico 2022.
E. COLI STEC MONITORING IN A ONE HEALTH APPROACH.
This PhD thesis investigates the epidemiology of Shiga toxin-producing Escherichia coli (STEC) in dairy herds, employing a One Health approach to enhance surveillance and public health interventions. Recognizing the significant public health threat posed by STEC, particularly its association with severe gastrointestinal diseases and hemolytic uremic syndrome (HUS), this research aims to provide a comprehensive understanding of the pathogen's dynamics across human, animal, and environmental interfaces.The introduction outlines the historical context of E. coli discovery and its classification into various serogroups and serotypes, with a particular focus on the pathogenic STEC strains that have emerged as critical public health concerns since the first recognized outbreak in 1982. The study emphasizes the importance of understanding virulence factors, such as Shiga toxins and the locus of enterocyte effacement (LEE), which contribute to the pathogenicity of STEC. The role of animal reservoirs, particularly ruminants, is highlighted as they serve as asymptomatic carriers, facilitating zoonotic transmission to humans.The methodology section details the rigorous sampling and molecular diagnostic techniques employed, including Real-Time PCR assays for detecting STEC serogroups. The study encompasses sample collection from various matrices, DNA extraction protocols, and validation of molecular methods to ensure reliability in detecting STEC strains. In-silico analyses on whole genomes retrieved from literature provide insights into the virulence and transmission dynamics of these pathogens.Results indicate a significant prevalence of STEC in dairy herds, with varying serotype distributions across different environments. The findings underscore the necessity for enhanced monitoring systems that integrate molecular diagnostics to identify high-risk populations and inform targeted interventions. The discussion contextualizes these results within existing literature, exploring implications for public health policies aimed at reducing STEC transmission from livestock to humans.In conclusion, this thesis contributes valuable knowledge to the field of public health sciences by elucidating the complex interactions between STEC, its animal reservoirs, and human health risks. The integration of a One Health perspective not only enhances understanding of STEC epidemiology but also advocates for collaborative efforts among veterinary, agricultural, and public health sectors to mitigate the risks associated with this pathogen. Future research directions are proposed to further investigate non-O157 STEC strains and their emerging threats to food safety and public health. This work lays a foundation for developing more effective surveillance strategies and interventions that can ultimately reduce the burden of STEC infections globally.
E. coli STEC monitoring in a One Health approach / V.m. Sora ; tutor: A. Zecconi ; coordinatore: C. La Vecchia. Dipartimento di Scienze Biomediche, Chirurgiche ed Odontoiatriche, 2024 Jan 28. 37. ciclo, Anno Accademico 2023/2024.
THE ROLE OF PLASTICS AS CARRIER OF ENVIRONMENTAL POLLUTANTS: AN INTEGRATED APPROACH TO ASSESS TOXICITY ON AQUATIC AND TERRESTRIAL SPECIES.
The uniqueness of plastic is given by its several advantages, including low cost, lightweight, flexibility and resistance. These properties exponentially increased the global demand for plastic in the last 70 years, turning this revolutionary material into a global environmental threat. Indeed, the mismanagement of plastic waste, most of which is still sent to landfills or directly littered in the environment by human improper behaviour, is becoming a serious threat for ecosystems. The effects of plastic dispersal in the environment potentially increase when considering the fragments of smaller dimension, which derive from the degradation of macroplastics or are intentionally produced for industrial applications. A lower dimension may therefore enhance the plastic fragment capacity to accumulate at tissue level, but also to act as vector for chemical contaminants. The project was aimed at evaluating the environmental impact due to the presence of micro- and nanoplastics, exposing aquatic and terrestrial species and observing the potential adverse effects using multi-tiered approaches. Several laboratory experiments were conducted using both freshwater model species, the vertebrate Danio rerio and the invertebrate Dreissena polymorpha, and, for the first time, the terrestrial model Bombyx mori. Moreover, the role of plastics as carrier of environmental contaminants was investigated on zebrafish larvae. All organisms showed the accumulation of plastics and their infiltration in different tissues. The biochemical analyses highlighted only slight effects of micro- and nanoplastic exposures, while more relevant results emerged from behavioural tests, which deserve further studies. Furthermore, the effects at individual level, combined with proteomic analyses, pointed out a shift in the toxicity of plastics and chemical contaminants when co- administered, suggesting a more complicated relationship then an additive or synergistic effect. Finally, the study paves the way for the use of B. mori larva as model organism for the evaluation of the effects of plastic particles on terrestrial ecosystems, which are still largely unknown.
THE ROLE OF PLASTICS AS CARRIER OF ENVIRONMENTAL POLLUTANTS: AN INTEGRATED APPROACH TO ASSESS TOXICITY ON AQUATIC AND TERRESTRIAL SPECIES / C.c. Parenti ; tutor: A. Binelli ; co-tutor: C. Della Torre ; coordinatore: F. Ficetola. Dipartimento di Bioscienze, 2021 Jan 29. 33. ciclo, Anno Accademico 2020. [10.13130/parenti-camilla-carla_phd2021-01-29].
ADVANCED AAV-MEDIATED LIVER-DIRECTED GENE THERAPIES FOR HAEMOPHILIA A AND MUCOPOLYSACCHARIDOSIS TYPE VI
Liver-directed gene therapy using adeno-associated viral (AAV) vectors holdssignificant promise for providing long-term transgene expression following asingle systhemic administration. However, both the limited cargo capacity (~4.7kb) and the non-integrative nature of AAV vectors prevent their application toconditions caused by either mutations in large genes (i.e. F8 whose codingsequence exceeds ~ 4.7 kb) or to those that require early stage of intervention,such as early-onset inborn errors of metabolism. My PhD project aims toovercome these shortcomings and broaden the therapeutic applications of AAVs.To overcome the issue of limited cargo capacity, I explored an AAV-mediatedprotein trans-splicing (PTS) strategy utilizing split inteins. PTS is a naturalmechanism employed by organisms across all domains of life to reconstitute largeproteins from shorter precursors. Split inteins facilitate this process by selfexcisingfrom the host protein in an energy-independent manner. I used thisapproach for haemophilia A (HemA), the most common X-linked bleedingdisorder (affecting 1 in 5,000 males), caused by a deficiency of clotting factor 8(F8, ~7 kb). I first demonstrated the successful full-length reconstitution of thelarge (~5 kb) and highly active B-domain deleted (B-DD) N6-F8 variant in vitro.Subsequently, I designed two separate AAV vectors, with regular size genomes,each encoding for half of the N6-F8, flanked by split inteins. The AAV-N6 inteinsystem led to successful reconstitution of functional full-length N6-F8 in themouse liver, achieving therapeutic F8 activity in haemophilic mice withouteliciting anti-F8 immune responses.In parallel, I developed an AAV-mediated CRISPR/Cas9 homology-independenttargeted integration (AAV-HITI) platform to achieve stable transgene integrationat the 3’ end of the mouse Albumin locus, which is highly expressed inhepatocytes. This approach mitigates the loss of episomal AAV genomes inproliferative tissues, thereby preserving therapeutic efficacy over time.14I demonstrated AAV-HITI efficacy, in mouse models of HemA andMucopolysaccharidosis type VI (MPS VI), a lysosomal storage disorder,achieving stable therapeutic levels of systemic proteins even at low AAV doses.Importantly, I performed comprehensive molecular analyses to characterize theAAV-HITI outcomes within the target site confirming successful targetedintegration. Moreover, in AAV-HITI treated livers nor gross chromosomalrearrangements or significant off-target insertions/deletions were observed 1-yearpost neonatal delivery. Furthermore, I demonstrated the effectiveness of thisplatorm in adult MPS VI and HemA mice, using vector doses that are within thesafe range for human translation.Overall, my findings support two distinct AAV-mediated liver-directed platformsto address major limitations of current AAV-based therapies. The AAV-inteinN6-F8 system holds promise as a potential therapeutic option for HemA, whilethe AAV-HITI platform could serve as a durable therapeutic option for livertargeteddiseases, particularly those requiring early intervention.These innovations represent significant advancements in overcoming challengesrelated to cargo capacity, stable transgene expression, and long-term efficacy ingene therapy.
ADVANCED AAV-MEDIATED LIVER-DIRECTED GENE THERAPIES FOR HAEMOPHILIA A AND MUCOPOLYSACCHARIDOSIS TYPE VI / F. Esposito ; tutor: A. Auricchio ; internal supervisor: N. Brunetti-Pierri ; external supervisor: A. Follenzi ; coordinator: S. Minucci. Dipartimento di Oncologia ed Emato-Oncologia, 2024 Dec. 36. ciclo, Anno Accademico 2023/2024.
ETERNAL SUNSHINE TO THE FAPLESS MASCULINITY:A DIGITAL ETHNOGRAPHY OF THE R/NOFAP COMMUNITY ON REDDIT PROMOTING ABSTENTION FROM PORNOGRAPHY CONSUMPTION AND MASTURBATION.
This study examines the NoFap phenomenon, a homosocial digital movement promoting abstinence from pornography and masturbation to overcome self-diagnosed porn addiction and “compulsive sexual behaviors”. Critical literature on the topic has identified heterosexist, patriarchal, and misogynistic discourses associated with this practice, dominated by evolutionary narratives on gender and sexuality, men’s sexual entitlement to women, and the imperative of heteronormative coital sex. Building on this literature, this research offers three key contributions to deepen our understanding of the widespread NoFap phenomenon and provide analytical tools for examining the reproduction of contemporary masculinities and the heteropatriarchy in digital spaces. First, NoFap is conceptualized as a Foucauldian technique of internalized biopower that seeks to discipline a body constructed as hetero-masculine, reinforcing regulatory ideals of hetero-masculinity and contributing to the (re)production of a heteropatriarchal society. Second, I introduce the concept of “affordances of the (gendered) self”, which conceptualizes the embodied use of affordances in r/NoFap as Foucauldian technologies of the self, integral to men’s self-improvement and self-optimization in pursuit of a post-masturbatory heteromasculinity. Third, I argue that r/NoFap can be understood as a form of a “diluted manosphere”, where manosphere content is “watered down”. In this context, NoFap presents itself as a space that appears to promote equality while still allowing participants to engage “safely” with ideologies, practices, and discourses that protect male privileges and uphold gendered power relations. This study is based on a year-long ethnographic investigation of r/NoFap, Reddit’s central hub for the movement, which had 1.1 million members in 2022. To navigate Reddit’s vast, real-time, networked, and algorithmically structured environment, a novel approach to digital ethnography was developed. This approach integrates computational tools within the ethnographic continuum, not to impose rigor on qualitative findings but to support, guide, and inform them throughout their production. By doing so, it subverts the platform’s algorithmic logics, ensuring that less visible content, suppressed by Reddit’s algorithms, is also considered.
ETERNAL SUNSHINE TO THE FAPLESS MASCULINITY:A DIGITAL ETHNOGRAPHY OF THE R/NOFAP COMMUNITY ON REDDIT PROMOTING ABSTENTION FROM PORNOGRAPHY CONSUMPTION AND MASTURBATION / C. Perin ; supervisor: R. Ferrero Camoletto ; co-supervisor: A. Caliandro ; director of doctoral program: A. Rebughini. Università degli Studi di Milano, 2025 May 05. 37. ciclo
SURGICAL TREATMENT OF ORAL LEUKOPLAKIA: A RANDOMIZED CONTROLLED CLINICAL TRIAL COMPARING SURGERY WITH WAIT AND SEE APPROACH
Rationale of the study: Different treatments have been proposed for the management of the most common oral potentially malignant disorders (i.e., leukoplakia and erythroleukoplakia) in order to prevent the risk of oral cancer. However, there is currently no consensus on the most effective approach for the patients affected by such lesions. Surgery is often performed, but in literature there is no randomized clinical trial that demonstrates its real effectiveness in preventing malignant transformation.Objective: To compare the incidence of oral squamous cell carcinoma (OSCC) in a group of subjects undergoing surgical treatment and a similar group under strict clinical and histological surveillance (“wait and see” approach). To evaluate the long-term recurrence rate in test group after surgical excision of the oral leukoplakia.Study Design: Multicentric randomized controlled clinical trial (RCT).Setting: Unit of Oral Medicine - ASST Santi Paolo e Carlo Milano - Head and Neck Department San Paolo Hospital of Milan - Università degli Studi di Milano - Reference Center of the study; Unit of Oral and Medicine and Oncology - CIR Dental School of Turin - Università degli Studi di TorinoPatients and Method: Inclusion criteria: adult patients to whom, after an incisional diagnostic biopsy, have been diagnosed an oral leukoplakia or erythroplakia. Exclusion criteria: patients affected by a previous oral cancer, patients under the age of 18, pregnant patients, patients affected by PVL, lesions located in anatomical risk areas in which the excision may cause post-surgical deficit (salivary ducts, nerves, arteries…) and lesions more extensive than 3cm.Patients included in the study are divided into two groups: surgery (test group) and wait and see (control group). The randomization has been created using an online tool.Demographics data, habits, clinical information are collected for both groups. Technique of excision, surgical margins, recurrence and intra and post-surgical complications, are collected in the test group, while clinical and histopathological changes are collected in the control group.Outcomes Primary: oral cancer incidence Secondary: time to malignant progression from diagnosis, recurrence rate in test group.Conclusion and clinical relevance: This study will compare surgical treatment with “wait and see” approach in order to evaluate the real effectiveness of surgery in preventing and reducing oral cancer incidence in a group of subjects affected by oral leukoplakia. This study will contribute to determine which is the most appropriate management for such patients and to establish a gold-standard treatment.
SURGICAL TREATMENT OF ORAL LEUKOPLAKIA: A RANDOMIZED CONTROLLED CLINICAL TRIAL COMPARING SURGERY WITH WAIT AND SEE APPROACH / N.g. Lombardi ; tutor: G. Lodi ; coordinator: M. Del Fabbro. Dipartimento di Scienze Biomediche, Chirurgiche ed Odontoiatriche, 2022 Dec 14. 35. ciclo, Anno Accademico 2022.
THE COOPERATION DILEMMA IN COUNTERTERRORISM OPERATIONS
"Which factors drive counterterrorism (CT) cooperation? Academic research has studied terrorism and counterterrorism since the seventies, yet such a question has remained unanswered. As terrorism was mainly considered an internal problem, scholars focused on domestic responses, while international strategies have received meagre attention. This work aims at answering this question, merging CT and Foreign Policy Analysis (FPA) literature. Two are the factors upon which the merger is built: threat perception prompted by terrorism and the domestic environment. By importing and combining elements from these fields, it is possible to reach a better explanation. To conduct the inquiry, I adopted a mixed method design approach. At first, I conducted a quantitative study on two newly collected datasets using panel data analysis. These datasets include CT bilateral agreements signed by European countries between 2002 - 2017. Then, I examined two case studies to test the robustness of the results of the first analysis. These cases have been analysed separately through process-tracing. The findings confirm the theoretical framework. Emotions are pivotal: both analyses show that when the perception of the threat is low, it is improbable that two countries sign a CT cooperation agreement. Similarly, the domestic environment does matter. Ideology of governments is crucial, corroborating what previous works underlined: parties in the centre of the left-right scale are more inclined to cooperate. On the other hand, findings also show that threat perception impacts cabinets at the extreme of the left-right scale, pushing them to cooperate with other nations. This work adds three new contributions to the field. First is the role of emotions in shaping international relations. Although scholars have argued the importance of this factor for domestic CT policies, studies on international CT cooperation have not fully considered this aspect. Furthermore, unlike previous works, this study compares the effect of threat perception with the effect of the actual impact of terrorism, showing that the former is much more important than the latter. The second contribution is the analysis of the role played by domestic structure on CT cooperation. Although in recent years many FPA and IR scholars have been examining the role of this factor, in CT studies such a ""domestic turn"" has yet to come. This thesis tries to open the field to a political perspective that can help reach a complete explanation of international CT cooperation. Lastly, this thesis fosters the study of counterterrorism in Europe. Scholars have utterly studied EU counterterrorism and the domestic policies adopted by its members. By contrast, they devoted less attention to the analysis of bilateral cooperation between EU members, even if it still is a fundamental part of their strategy against terrorism. Therefore, not only this work sheds light on an issue that has been poorly debated, but it does so by looking at a case - bilateral cooperation between EU members - that literature on EU CT has little considered so far."
THE COOPERATION DILEMMA IN COUNTERTERRORISM OPERATIONS / F. Baraldi ; tutor: F. Coticchia ; coordinatore dottorato: M. Jessoula. Università degli Studi di Milano, 2022 Nov 16. 34. ciclo, Anno Accademico 2021.
ETHNOBOTANICAL SURVEYS IN LOMBARDY (ITALY): A MULTIDISCIPLINARY APPROACH TO STUDYING THE PAST TO IMPROVE THE FUTURE.
In the thesis presented herein, three case studies, with focus on ethnobotanical investigations conducted in Lombardy (North of Italy), are described. Considering ethnobotany as a multidisciplinary and cyclic science, the main phases of an investigation (the field, the laboratory, and the return to the population phases) are considered and the case studies are designed to cover them all.The case studies are the following:1. Case study Milan: a peculiar historical investigation on the majolica jars preserved at the Ca' Granda Ospedale Maggiore Policlinico, once used to store plant ingredients and medicinal remedies administered to the patients of the hospital. The thorough analysis of the inscriptions of the jars and extensive research on literature of the time allowed to hypothesize the medicinal species once potentially cultivated in the Hortus simplicium (Garden of Simples) of the hospital. Research in current scientific literature represented a starting point for the theoretical validation or rejection of the ancient remedies of the jars.2. Case study Valle Imagna (Bergamo): the starting point of this case is the previous analysis of an ancient 18th century manuscript containing medicinal remedies. The diachronic comparison of the medicinal uses retrieved from historical sources with the traditional ones recorded during an ad hoc ethnobotanical investigation conducted on the territory of upper Valle Imagna allowed for the selection of a plant species symbol of the territory. The subsequent a multi-step laboratory survey was aimed at the thorough examination of the selected species, Agrimonia eupatoria L. (Rosaceae), from a micromorphological, phytochemical, and pharmacological standpoints.3. Case study Alpine Lombardy: the case was developed along different Alpine territories: Sondalo (Valtellina, Sondrio), Valmalenco (Sondrio), and the neighboring Valposchiavo (Canton of Grisons, Switzerland). The field investigations of the three territories are presented. Additionally, the case involved the laboratory phase conducted in continuity with a previous PhD project in Valmalenco on the species Achillea erba-rotta subsp. moschata (Wulfen) I.Richardson and the nutritional analyses on six wild edible species collected in Valmalenco. Finally, different strategies for the return to the population phase are outlined and discussed.Finally, in the Appendix attached to the main thesis, all the activities of dissemination of the scientific results and third mission of the three-year PhD project are thoroughly described.
ETHNOBOTANICAL SURVEYS IN LOMBARDY (ITALY): A MULTIDISCIPLINARY APPROACH TO STUDYING THE PAST TO IMPROVE THE FUTURE / F. Milani ; tutor: G. Fico ; phd coordinator: G. Vistoli. - DISFARM - UNIMI - Milano. Dipartimento di Scienze Farmaceutiche, 2025 Jan 29. 37. ciclo, Anno Accademico 2023/2024.
INTRINSIC AND EXTRINSIC DRIVERS OF FORAGING MOVEMENTS IN COLONIAL BIRDS
Foraging is a central trait in species’ life history, being tightly linked to individual fitness and therefore to population processes. To be successful, individuals should forage in a way that minimize energy expenditure and maximize energy intake. However, this balance depends on a mixture of different elements of intrinsic and extrinsic nature, that drive foraging choices of individuals.Intrinsic drivers are elements inextricably linked to individual’s characteristics or qualities that often are a function of age or sex (e.g., dimorphism, social dominance). Extrinsic drivers are elements linked to the surrounding environment, that can be abiotic (e.g., landscape features, weather), or biotic (e.g., heterospecific/conspecific presence, resources distribution). In this thesis I used a combination of bio-logging technologies and statistical modelling techniques to investigate in detail the drivers of foraging movements in different colonial bird species. Specifically, I investigated intrinsic and extrinsic drivers on sea and land (Chapters I and II), the effects of competition (Chapter III), the role of weather (Chapter IV), how food resources can be found in the sea (Chapter V) and its consequences (Chapter VI). My results suggest that a complex interplay of intrinsic and extrinsic drivers shape spatio-temporal foraging decisions in wild bird species, and their combined effect can be sometimes difficult to disentangle. Wind conditions, rainfall, morphological differences linked to sex, inter- and intra-specific competition all affect how individuals optimize their search for food, via the adoption of different behavioural tactics. Moreover, the search for food is particularly challenging in the marine environment, that is extremely dynamic and three-dimensional. Here, individuals could use a combination of static and dynamic features to locate prey aggregations. However, such features also aggregate contaminants such as microplastic, enhancing the risk of ingestion. In conclusion, understanding the connection between individual qualities, foraging movements and external agents is particularly important given the predicted global changes for future years. Ultimately, behavioural flexibility in foraging could be an important trait to successfully cope with such changes and could contribute to increase the resilience of populations over time.
INTRINSIC AND EXTRINSIC DRIVERS OF FORAGING MOVEMENTS IN COLONIAL BIRDS / F. De Pascalis ; tutor: D. Rubolini ; co-tutor: J. G. Cecere. Dipartimento di Scienze e Politiche Ambientali, 2022 Apr 29. 34. ciclo, Anno Accademico 2021.
TUMOR MICROENVIRONMENT IN EXPERIMENTAL PRECLINICAL MOUSE MODELS OF HUMAN CANCER: MORPHOLOGICAL APPROACH
One of the recent advancements in oncological research has been the recognition of the tumor microenvironment (TME) as a relevant participant during all stages of the evolution of a neoplastic process. Indeed, over the past decades, tumors have been considered through a changing perspective: no longer as a growth of homogeneous neoplastic cells, but as an actual organ composed of different cell populations and structures: the parenchyma being the neoplastic population and the stroma, including the vascular network and infiltrating cells. The tumor microenvironment has a dual role in tumor biology, both promoting and antagonizing tumor development, growth, and local or distant invasiveness. According to its leading role in influencing tumor biology each component of the TME could be considered as a potential pharmacological target to be enhanced or antagonized, in order to influence tumor behavior. Accordingly, the study of the TME could provide new insights in the tumor biology and offers numerous potential targets for the development of novel therapeutic strategies. In this context, morphological techniques represent useful tools for the investigation of the TME, allowing the evaluation of the spatial distribution of the different elements, and provide useful complementary information to clinical and other data obtained in experimental in vivo studies. In this thesis, the three main classes of the TME components -tumor-associated vasculature, immune-inflammatory cells and tumor stroma- are illustrated in three different chapters and relevant experimental studies described. However, it should be considered that the various aspects of TME are not separate entities but are all involved in a dynamic system with complex structural and functional interactions. Chapter 1 – Tumor-associated vasculature Tumor angiogenesis has been identified as a hallmark of cancer, due to its central role in supporting tumoral growth, providing nutrient supply, removing catabolites and enabling tumoral metastatic dissemination. Most of the solid tumors are characterized by an “angiogenetic switch” in which an imbalance between pro- and anti-angiogenic factors sustains a dysregulated angiogenetic process, leading to the formation of an altered vascular network composed of structurally and functionally abnormal blood vessels. Drugs targeting tumor vasculature has been extensively studied as a mean to interfere with tumoral growth as well as to promote the delivery and/or effect of co-administered compounds to the tumor. In the first study of this chapter, we demonstrated the therapeutic efficacy and the antiangiogenic effect of a novel compound developed by binding sunitinib (a well-known antiangiogenic drug) to a selective binder of αVβ3 integrin thus promoting its delivery to the target site (tumors expressing αVβ3 integrin). The other studies of this chapter investigated the relation between tumor vasculature and tumor hypoxia. In particular, this relation was investigated to uncover the potential mechanism underlying the synergistic effect of the administration of an antiangiogenic compound (cediranib) with a poly-ADP ribose polymerase (PARP) inhibitor (olaparib) in a panel of patient-derived xenografts of ovarian carcinoma. Chapter 2 – Tumor immune microenvironment In most cancers, both innate and acquired immunity have a driving role during all stages of tumor development and progression. Depending on the cell population and/or molecular stimuli received, they can act in a dual way, antagonizing or promoting tumor growth. Three selected studies were described in chapter 2 and investigated: 1. The role of NK cells in hindering metastasis engraftment in a metastatic model of synovial sarcoma. After the combined administration of an heparanase-inhibitor with a tyrosine kinase inhibitor a significant reduction of lung metastases was observed and immunohistochemical analyses demonstrated the role of NK
TUMOR MICROENVIRONMENT IN EXPERIMENTAL PRECLINICAL MOUSE MODELS OF HUMAN CANCER: MORPHOLOGICAL APPROACH / L. Minoli ; tutor: E. Scanziani ; coordinatore corso di dottorato: V. Grieco. DIPARTIMENTO DI MEDICINA VETERINARIA, 2020 Feb 05. 32. ciclo, Anno Accademico 2019. [10.13130/minoli-lucia_phd2020-02-05].
ACCELLERATED SELF-LIFE TESTING OF COFFEE: THE ROLE OF INNOVATIVE FLEXIBLE PACKAGING MATERIALS ON OXIDATION AND SHELF-LIFE ESTIMATION WITH A SIGHT ON THEIR ENVIRONMENTAL SUSTAINABILITY.
The preservation of roasted and ground coffee quality during storage represents a critical challenge for the food packaging industry. Coffee's unique sensory and chemical profile is highly sensitive to environmental conditions, including oxygen, humidity, and temperature. These factors accelerate degradation processes, such as oxidation, leading to a decline in aroma, flavour, and shelf life. The development of effective, sustainable, and high-performance packaging solutions requires a multidisciplinary approach that combines material science, analytical techniques, sensory evaluation, and environmental sustainability. This dissertation addressed these challenges by exploring innovative solutions across five interconnected studies, providing actionable insights for the coffee packaging industry.The research began with an in-depth analysis of gas and moisture permeation in polyolefin-based packaging structures. Seven flexible formats, incorporating ethylene vinyl alcohol (EVOH) and metallized polypropylene barrier layers, were evaluated under varying temperature and relative humidity conditions. The findings revealed that EVOH-based materials are more sensitive to environmental changes due to their hydrophilic nature, while metallized structures exhibited superior stability. The positioning of the barrier layer within multilayer architectures was optimized using response surface methodology, achieving a minimal layer distance of 58 µm for optimal oxygen permeability under high-stress conditions. These insights were applied to estimate the two-year shelf life of coffee packages, identifying metallized and select EVOH-based materials as the most effective solutions.The second focus of this work involved the evaluation of advanced analytical techniques such as electronic nose (e-nose), gas chromatography-mass spectrometry (GC-MS), and Fourier-transform near-infrared (FT-NIR) spectroscopy, for monitoring coffee aging. These methods were combined with sensory analysis to investigate the evolution of coffee quality under adverse conditions. Sensory evaluation confirmed a decline in attributes such as aroma intensity and aftertaste, accompanied by increases in bitterness and oxidation-related characteristics. Analytical techniques identified key markers of degradation, including volatile organic compounds (VOCs) and chemical changes in oily compounds and carbohydrates. Integrating these data with principal component analysis (PCA) provided a comprehensive understanding of coffee quality evolution during storage.Oxidation was further investigated through a study on the impact of high oxygen concentrations and varying storage temperatures. Results demonstrated that oxygen consumption and sensory oxidation attributes follow distinct thermal sensitivities, with oxidation thresholds ranging from 330 µg to 430 µg of oxygen per gram of coffee. These findings enhance the ability to predict shelf life and optimize packaging solutions under real-world conditions.The final component of the research examines the environmental impact of coffee packaging structures using Life Cycle Assessment (LCA). Comparing traditional aluminium-based materials with recyclable mono-materials, the study revealed that metallized recyclable packaging offers the best balance between barrier performance and sustainability. The results advocated for reducing raw material use and increasing recycling rates to align with circular economy principles.By integrating material performance, analytical monitoring, sensory evaluation, and sustainability assessments, this dissertation provided a holistic framework for addressing the complex demands of coffee packaging. The findings contributed to the development of innovative, efficient, and environmentally responsible packaging solutions, meeting the dual objectives of preserving coffee quality and reducing environmental impact. This work underscored the importance of m
ACCELERATED SHELF-LIFE TESTING OF COFFEE: THE ROLE OF INNOVATIVE FLEXIBLE PACKAGING MATERIALS ON OXIDATION AND SHELF-LIFE ESTIMATION WITH A SIGHT ON THEIR ENVIRONMENTAL SUSTAINABILITY / F. De Agostini ; tutor: S. Limbo ; co-tutor: C. Alamprese ; coordinatore: D. Mora. Dipartimento di Scienze per gli Alimenti, la Nutrizione e l'Ambiente, 2025 Mar 03. 37. ciclo, Anno Accademico 2024/2025.
LA PRESSE CLANDESTINE DE LA RÉSISTANCE FRANÇAISE (1941-1944) : STYLES, LANGAGES, RHÉTORIQUE ET ARGUMENTATIONS
Studies conducted to date on the French Resistance and the closely related clandestine press have been based on an analysis of the themes conveyed by Resistance periodicals and the way in which clandestine newspapers were produced and distributed during the Occupation. However, little attention has been paid, either historically or linguistically, to the primary nature of the Resistance's clandestine press and what it really represents: a medium. This thesis project is part of the field of linguistic studies and aims to focus on the linguistic, stylistic, rhetorical and argumentative strategies used in clandestine periodicals to disseminate content that contrasts with the dominant official discourse. The aim of this thesis is therefore to place itself in the intermediate space between the major historical studies of the French Resistance and the existing analyses of the clandestine press, by proposing a contribution to what has not yet been sufficiently considered. The analysis of the linguistic, argumentative and rhetorical strategies opens the possibility of completing from a linguistic point of view, insufficient to date, the studies on memory and the Resistance. The study of the as yet little-explored field of language also provides a better understanding of the aims of the Resistance and the contradictions that characterised one of the most troubled periods in contemporary history, and thus makes a necessary contribution to a subject that continues to occupy a prominent place in France's collective memory.
LA PRESSE CLANDESTINE DE LA RÉSISTANCE FRANÇAISE (1941-1944) : STYLES, LANGAGES, RHÉTORIQUE ET ARGUMENTATIONS / A. Della Rocca ; tutor: M. Barsi, A.-R. Hermetet. - Università degli Studi di Milano. Dipartimento di Lingue, Letterature, Culture e Mediazioni, 2024 Mar 26. 36. ciclo, Anno Accademico 2023.
INVESTIGATING THE BASES OF THE SPECIFICITY OF THE RESTRICTOR COMPLEX FOR THE TERMINATION OF NONCODING TRANSCRIPTION
Rampant transcription initiation occurring at the tens of thousands of cis-regulatory regions in the genome can be detrimental to the normal functioning of the cell. Several transcription termination mechanisms act to keep the initiation of non-productive transcription in check. The Restrictor complex, composed of the adaptor protein WDR82 and the RNA-binding protein ZC3H4, suppresses processive transcription of thousands of noncoding RNAs (ncRNA) in mammalian genomes. The mechanism of Restrictor-mediated termination is not known but does not involve cleavage of the nascent RNA, setting it apart from known mammalian termination machineries. Furthermore, it is uniquely specific for noncoding transcription.Here, we show that ZC3H4 binds its non-coding RNA targets with high specificity, thereby bringing about Restrictor termination activity. We show that ZC3H4 RNA binding occurs at the very 5’ end of the transcript, to a specific RNA consensus sequence. We found that the activity of the Restrictor complex is more widespread than previously thought, with its depletion resulting in processive transcription at a vast number of previously undetectable ncRNAs. Further, we show that PNUTS, a protein implicated in transcription termination and also in the Restrictor-mediated pathway, directly binds to nascent RNA originating from different genomic loci where PNUTS’ phosphatase binding partner PP1 is known to act. We show that among the PNUTS-bound RNAs are Restrictor-sensitive ncRNAs, thereby providing a basis for the Restrictor-PNUTS synergy recently reported. Finally, I show evidence of sequence specificity of PNUTS’ RNA binding, suggesting that the underlying RNA sequence facilitates recruitment of PNUTS-PP1 for targeted dephosphorylation events.
INVESTIGATING THE BASES OF THE SPECIFICITY OF THE RESTRICTOR COMPLEX FOR THE TERMINATION OF NONCODING TRANSCRIPTION / G.m. Mandana ; tutor: G. Natoli ; internal advisor: B. Amati ; co-tutor: D. Pasini. Università degli Studi di Milano, 2025 Jan 21. 36. ciclo, Anno Accademico 2024.
THE INTEGRATED PROTECTION OF FUNDAMENTAL RIGHTS IN THE EUROPEAN CONSTITUTIONAL LAW PERSPECTIVE
The research falls within the context of the Europe of rights and aims to assess the degree of maturity of integrated protection of fundamental rights, between national legal systems and the European legal framework. The study is based on the constant fluctuations between political Europe and the Europe of Courts, which have generated both advantages and uncertainties and imbalances in the system of fundamental rights protection.In order to deepen the research on integrated protection, we will pose two distinct questions. First, we intend to investigate the integrated protection of fundamental rights that arises from the inter-system relationships between the European Union and the Member States. Secondly, the second question aims to examine the degree of integration and/or cooperation between constitutional jurisprudence and that of the Court of Justice in the protection of rights based on both national Constitutions and the Charter of Nice.The scientific methodology that will be employed during the research is typical of European constitutional law, which focuses on the close relationship between the constitutional realities of the Member States and their interactions within the European Union.The conclusion of the research suggests that in order to overcome the chronic imbalances between the Europe of Judges and political Europe, it is necessary to take real steps forward in the process of constitutionalization of the Union.
THE INTEGRATED PROTECTION OF FUNDAMENTAL RIGHTS IN THE EUROPEAN CONSTITUTIONAL LAW PERSPECTIVE / P. Gambatesa ; tutors: M. D'Amico, A. Aguilar Calahorro, C. Del Bò ; coordinators: L. Sacconi, A. Aguilar Calahorro. Dipartimento di Diritto Pubblico Italiano e Sovranazionale, 2024 Apr 22. 36. ciclo, Anno Accademico 2022/2023.
LA MOBILITAZIONE DEI FAMILIARI DI DESAPARECIDOS IN MESSICO E LA ¿GUERRA AL NARCOTRAFFICO¿. IL CASO DEL COLLETTIVO FUUNDEC-M.
The purpose of this thesis is to investigate the conditions that encourage the activism of victims’ relatives in high-risk contexts. It will be possible through the analysis of the process of mobilization of these kind of social actors. Based on sociological literature of social movements, the research aims to give a multifactorial response to this peculiar collective action. Three dimensions of analysis will be taken into consideration: the socio-political context (macro), the actors and networks (meso) and the individual (micro). This work focuses the attention on a single case study: Fuerzas Unidas por Nuestros Desaparecidos en Coahuila y en México – FUUNDEC-M. This collective of relatives of forced disappearances’ victims was born and developed in the North-eastern Mexican state of Coahuila, a region characterized by the presence of organized crime and high levels of violence, impunity and political-institutional corruption.
LA MOBILITAZIONE DEI FAMILIARI DI DESAPARECIDOS IN MESSICO E LA ¿GUERRA AL NARCOTRAFFICO¿. IL CASO DEL COLLETTIVO FUUNDEC-M / T. Aureliani ; tutor: F. dalla Chiesa ; co-tutor: M. Massari; coordinatore: F. dalla Chiesa. Dipartimento di Studi Internazionali, Giuridici e Storico-Politici, 2020 Jun 18. 32. ciclo, Anno Accademico 2019. [10.13130/aureliani-thomas_phd2020-06-18].
SPROUTING AS A BIOTECHNOLOGICAL PROCESS TO IMPROVE THE FUNCTIONAL PROPERTIES OF CEREAL-BASED PRODUCTS
Although some studies showed that the use of flours from controlled sprouted grain can improve bread properties, the relationship between macromolecule functionality and bread features needs to be investigated, especially in the case of fiber-enriched products. In this context, the PhD project focused on the assessment of the sprouting effects on the functionality of the main macromolecules in grains (i.e., protein, starch, and fiber) and their impact on dough and bread-making properties.In the introduction chapter (Chapter 2), a literature review has been carried out, where the main effects of sprouting on the nutritional profile of grains and their health benefits were summarized. Moreover, the scaling-up of the sprouting process from domestic to industrial level, the importance of monitoring processing parameters were addressed.The first two sections (Section 4.1 and 4.2) of the Results and Discussion chapter assessed the effects of the sprouting process on technological/functionality of wholegrain flours from common wheat. In particular, wholegrain flours were obtained from single-stream milling (Section 4.1) and the recombination approach by including 20% bran to wheat flour (Section 4.2). Sprouting process caused a worsening in both gluten aggregation kinetics – suggesting a gluten weakening – and starch pasting and gelation properties. However, following the conditions obtained by the Farinograph test, it was possible to obtain a wholegrain bread with improved characteristics in terms of volume and crumb softness.Considering the positive effect of sprouting on the bread-making performance of common wheat, the process was carried out on durum wheat (Section 4.3), with the aim of improving its gluten functionality for the production of durum wheat bread. For this reason, the effects of enzymatic activities developed during sprouting on durum wheat kernel characteristics, starch and gluten behavior were investigated, as well as their relationship with the bread-making performance. Although the sprouting process caused a decrease in the gelatinization and retrogradation capacity of starch and gluten aggregation properties, bread with improved volume and crumb porosity was obtained. In particular, the best results were obtained using wheat sprouted up to 48 h.The last part of the project focused on the sprouting of quinoa (Section 4.4) and sorghum (Section 4.5), whose consumption has been constantly increasing, since they are considered environmentally sustainable crops with high nutritional value. However, from the technological standpoint, the suitability of quinoa and sorghum for achieving good bread-making performance in wheat-based formulation still needs deep investigation. As regards enriched-bread, the presence of sprouted grains at 20% replacement level showed higher specific volume and lower crumb softness compared to control. In addition, in the case of sorghum, sprouting improved the in vitro protein digestibility.In conclusion, this PhD project provides evidence of how protein and starch functionality is affected by both the type of grain used and sprouting duration. In addition, bread-making performance of wheat-based formulations can be enhanced by using sprouted grains or their main milling fractions (refined flour or bran), opening a new approach to the production of fiber- enriched baked goods.
SPROUTING AS A BIOTECHNOLOGICAL PROCESS TO IMPROVE THE FUNCTIONAL PROPERTIES OF CEREAL-BASED PRODUCTS / G. Cardone ; tutor: A. Marti; coordinatore: A. Pagliarini. Dipartimento di Scienze per gli Alimenti, la Nutrizione e l'Ambiente, 2021 Dec 03. 34. ciclo, Anno Accademico 2021. [10.13130/cardone-gaetano_phd2021-12-03].
INVESTIGATING THE BIODIVERSITY IN SPONTANEOUS FERMENTATIONS AS A SOURCE OF HIGH-PERFORMANCE MICROORGANISMS
This PhD project was aimed at the investigation of spontaneously fermented food products with the goal of selecting highly performing strains to be used as starter cultures, in order to optimize the fermentation process and give to the final product an added value from a technologic, safety or sensory point of view.This project is divided in two parts dealing with two different fermentation processes. The first part is focused on sourdough fermentation and its potential to improve the baking performance of alternative grains such as spelt. Firstly, the dominating lactic acid bacteria (LAB) and yeasts were isolated and studied for their performance in fermentation as well as resistance to stress conditions. From the selected strains, one strain of Pediococcus pentosaceus was selected as a potential starter. To design a mixed starter culture, other LAB and yeast strains isolated from cereal fermentations were studied. First, we studied two LAB species, Weissella cibaria and W. confusa, that have interesting pro-technologic characteristics such as a good ability to produce exopolysaccharides, to degrade arabinoxylans and to inhibit the growth of some mold species. On the other hand, the two species have raised concerns as potential opportunistic pathogens. For this reason, combined genotypic and phenotypic studies were performed to assess the safety of the two species. Subsequently, an alternative non-Saccharomyces yeast species, Kazachstania unispora, was investigated as a leavening agent. The species had a good resistance to stress conditions and a good ability to ferment glucose, but lacked the ability to utilize maltose, the main carbon source in dough, for this reason it performed a slow leavening depending on the LAB to release free glucose. The selected strains were applied to a spelt-based sourdough bread with promising results; the LAB increased the mold-free shelf-life and improved the texture and conservation of the product, that received a high liking score in the sensory evaluation. The second part of the project regarded the cocoa fermentation and the possibility of using selected LAB as adjunct cultures to improve the quality of the chocolate in a controlled fermentation setup. Firstly, LAB and yeast strains were isolated and identified from natural cocoa fermentations. Two strains belonging to the species Lactiplantibacillus fabifermentans and Furfurilactibacillus rossiae were selected for further studies. We evaluated their adaptation to conditions associated with cocoa fermentations, such as low pH, high temperatures and high osmotic pressure as well as their ability to grow in conjunction with yeasts and acetic acid bacteria, whose activity is essential for cocoa fermentation. The two strains were then used as adjunct cultures in cocoa fermentations and the quality and sensory profile of the chocolate obtained was evaluated. This work allowed us to highlight the potential of autochthonous microbial strains with high level adaptation to the food matrix that allow not only to obtain a successful and reliable fermentation but to add to the safety, taste and quality of the final product.
INVESTIGATING THE BIODIVERSITY IN SPONTANEOUS FERMENTATIONS AS A SOURCE OF HIGH-PERFORMANCE MICROORGANISMS / D. Korcari ; tutor: M. G. Fortina ; phd coordinator: D. Mora. Dipartimento di Scienze per gli Alimenti, la Nutrizione e l'Ambiente, 2022 Mar 02. 34. ciclo, Anno Accademico 2021.
TARGETING TYPE-I PRMTS RESTORES PLATINUM SENSITIVITY IN OVARIAN CARCINOMA THROUGH MITOCHONDRIAL METABOLISM
Adaptive chemoresistance remains a critical barrier to effective ovarian cancer therapy, necessitating a deeper understanding of the cellular and metabolic changes that drive platinum resistance. Here, we dissect the interplay between platinum-induced genotoxic stress, type-I protein arginine methyltransferases–mediated post-translational modifications, and mitochondrial metabolic plasticity underpinning chemoresistance in EOC. We present an integrative multi-omics analysis of carboplatin-resistant A2780 ovarian cancer cells (A2780R) developed via stepwise drug adaptation, revealing pronounced mitochondrial and metabolic rewiring. A2780R cells display an 8.8-fold increase in carboplatin IC50 compared to parental A2780S cells and elevated mitochondrial respiratory capacity, and a hyperfused mitochondrial network, as quantified by 3D-skeleton imaging. Notably, pharmacological inhibition of Type-I protein arginine methyltransferases (PRMTs) with MS023 re-sensitizes A2780R cells to carboplatin, reducing IC50 by up to 56%, and abrogates the enhanced mitochondrial activity. MS023 treatment induces depolarization of the mitochondrial membrane, diminishes mitochondrial volume and network integrity, and triggers mitophagy.Proteomic and metabolomic analyses at steady state and upon PRMT inhibition revealed that A2780R cells adapt by favouring anabolic pathways, with notable upregulation of nucleotide and glutathione metabolism and increased amino acid catabolism. This metabolic rewiring is associated with elevated NADPH production from the pentose phosphate pathway, supporting enhanced glutathione-mediated redox buffering. MS023 disrupts these adaptations, depleting glutathione pools and precursor amino acids, reducing redox balance, and shifting central carbon metabolism. Isotope tracing confirms that A2780R cells channel serine-derived carbons into glutathione synthesis, a process blunted by Type-I PRMT inhibition. Mechanistic investigations indicate that R-methylation of mitochondrial proteins modulates biogenesis, metabolic flexibility, and oxidative stress buffering, establishing Type-I PRMTs as essential regulators of chemoresistance-associated metabolic shifts. Our findings position Type-I PRMT inhibition as a promising combinatorial approach to overcome platinum resistance by targeting maladaptive metabolic features in ovarian cancer cells. This study provides a comprehensive blueprint of the mitochondrial and metabolic phenotypes underpinning adaptive chemoresistance, highlighting the translational potential of metabolic vulnerabilities in the therapeutic sensitisation of resistant cancer phenotypes.
TARGETING TYPE-I PRMTS RESTORES PLATINUM SENSITIVITY IN OVARIAN CARCINOMA THROUGH MITOCHONDRIAL METABOLISM / A. Mondal ; tutor: T. Bonaldi ; co-tutor: N. Mitro. Dipartimento di Oncologia ed Emato-Oncologia, 2025 Dec 17. 37. ciclo, Anno Accademico 2024/2025.
SYMBIOTIC CHIMERAS: NOVEL IMMUNOLOGICAL PROPERTIES OF ASAIA SYMBIONTS ENGINEERED TO BE COVERED BY THE WOLBACHIA SURFACE PROTEIN
Vector-borne diseases represent one-sixth of all infectious diseases and cause more than 1,000,000 deaths every year. The World Health Organization (WHO) proposes the adoption of sustainable integrated vector management, which includes strategies such as environmental remediation, information and education of the population, but also the integration of classical chemicals (e.g. insecticides) with non-chemical control methods for the control of arthropod vectors. Among these strategies, the symbiotic control (SC), which exploits microorganism symbionts of the insects, is getting more and more attention as it is particularly promising for the control of vector-borne diseases.Particularly, in this thesis, I studied the potentiality of bacteria belonging to two genera: Asaia and Wolbachia. The former includes acetic acid bacteria, symbionts of many insects such as mosquitoes and recently proposed as good candidates for paratransgenesis-based control strategies. The latter includes symbionts of arthropods and nematodes, with the capability to stimulate innate immune responses in mosquitoes with a reduction of their vectorial capability. We thus engineered the bacterium Asaia to express the Wolbachia surface protein (WSP), a potential inductor of innate immunity, derived from Wolbachia infecting the nematode Dirofilaria immitis (AsaiaWSP).My project has two main aims:i) to test the capability of the bacterium AsaiaWSP to stimulate the immune system of mosquitoes and thus to interfere with D. immitis development after infection;ii) to verify if AsaiaWSP is able to induce a macrophage polarization during the immune response towards the M1/Th1 phenotype and if this polarization could determine an anti-Leishmania effect.The obtained results to achieve these aims have been summarized in two articles:Article 1The first study, here presented, consolidates the previous evidence on the immune-stimulating property of WSP. In fact, WSP from Wolbachia of D. immitis was shown to stimulate the immune response in mosquitoes and mammals (humans, rodents, dogs) in vitro, but here for the first time an engineered bacterium expressing WSP was tested. AsaiaWSP activated the expression of immune genes coding for effector molecules in Ae. aegypti and An. stephensi mosquitoes. Once verified the stimulation of the immune system of the mosquitoes, the capability to inhibit the development of the nematode D. immitis, ethiological agent of dirofilariasis, in the vector Ae. aegypti was investigated. We obtained a first evidence of an inhibition of D. immitis larval development after the infection of mosquitoes with the engineered bacterium AsaiaWSP.Article 2In the second paper, I focused my attention on leishmaniases, vector-borne diseases widespread in more than one hundred countries in tropical, sub-tropical and temperate zones and caused by the protozoan parasite Leishmania. I investigated the capability of the chimeric bacterium AsaiaWSP to polarize the immune response towards the M1/Th1 phenotype, which is protective for the host; in fact, macrophage polarization towards the M1/Th1 or M2/Th2 side is crucial for the outcome of the visceral leishmaniasis. The chimeric bacterium AsaiaWSP acted as a polarizing agent, stimulating the phagocytosis and inducing the release of M1/Th1 cytokines, ROS and the expression of iNOS. Then, AsaiaWSP determined an anti-leishmanial effect with a reduction of the number of intracellular parasites.In conclusion the modified bacterium Asaia, here proposed, appears as a promising candidate for paratransgenesis-based control strategies for the control of vector-borne diseases in general, and as a successful immunomodulator, which could be used in combination with the classical chemotherapeutic agents for the treatment of leishmaniases and other M1-impaired diseases.
SYMBIOTIC CHIMERAS: NOVEL IMMUNOLOGICAL PROPERTIES OF ASAIA SYMBIONTS ENGINEERED TO BE COVERED BY THE WOLBACHIA SURFACE PROTEIN / I. Varotto Boccazzi ; scientific tutor: S. Epis. Dipartimento di Bioscienze, 2020 Feb 21. 32. ciclo, Anno Accademico 2019. [10.13130/varotto-boccazzi-ilaria_phd2020-02-21].
WORD USE AND LANGUAGE STYLE MATCHING IN PATIENT-DOCTOR COMMUNICATION IN ONCOLOGY: PRESENT AND FUTURE
Patient-doctor communication is a central process in healthcare. It is related to various beneficial effects, as patient satisfaction with consultation and the implementation of a shared decision-making process. It has specific characteristics in the oncological context, and in new online-related contexts. Due to its importance, patient-doctor communication has been widely studied, with the majority of these studies focused on broad themes and content of the communication process. Less studied are the more-granular characteristics, as word use and verbal mimicry. With the diffusion of computerized text analysis methods, a growing literature is showing that the words people use daily have various relationships with psychological variables, while verbal mimicry, established between two people talking or writing to each other, is related to various relationship outcomes. LIWC, a computerized text analysis program, allows the analysis of word use and verbal mimicry, through LSM measure. This thesis work had the aim to analyze patient-doctor communication in its granular components. I aimed to analyze word use and LSM, established in the dyad patient- physician and in association with patient satisfaction with the consultation, in an online Italian, oncological setting. Moreover, the second aim was to lay the foundations to explore these constructs, and the shared decision-making process, in a real, face-to-face, Italian oncological setting.First, I started focusing on a more general context (Study 1). I analyzed the characteristics of word use and LSM in patient-healthcare professional communication in an English language “Ask-the-Expert” website. In this context, patients (i.e., users) can post a question that will be answered by a healthcare professional. I found different patterns of words used by patient and healthcare professionals, and medical doctors. Moreover, I found healthcare professionals’ and websites’ characteristics related to LSM. After these initial insights, I moved on a preparatory step, translating into Italian language the function word categories of the LIWC 2015 English Dictionary, and developing the Italian Function Words Dictionary (Study 2a). This instrument allowed to calculate LSM with Italian samples. I implemented it in a case study (Study 2b). Then, in Study 3, I analyzed word use, LSM, and the relationship between LSM and patient satisfaction with the consultation in an Italian, oncological “Ask-the-Expert” websites. I found specific patterns of word use in patients and oncologists, and an association between LSM, established between patients and oncologists, and patient satisfaction with the consultation. In Study 4, a general population study focused on a dyadic collaborative negotiation task. I found that LSM, established during this task, had a relationship with the perception of a shared decision-making process happened during the task, but only for some participants. These results laid the foundations to develop a study assessing these constructs – use of words, LSM, patient satisfaction with the consultation and shared decision-making process – in patient-doctor communication happening in real, oncological consultations. Moreover, these results are initial insights on these themes, that can have potential practical implication and shed new light on patient-doctor communication process.
WORD USE AND LANGUAGE STYLE MATCHING IN PATIENT-DOCTOR COMMUNICATION IN ONCOLOGY: PRESENT AND FUTURE / L. Vergani ; internal advisor: D. Monzani ; tutor: K. Mazzocco ; phd coordinator: S. Minucci. Dipartimento di Oncologia ed Emato-Oncologia, 2022 Mar 14. 33. ciclo, Anno Accademico 2021.
SCIENTIFIC CROSSCURRENTS BETWEEN ITALY AND ENGLAND: ITALIAN CONTRIBUTIONS TO THE PHILOSOPHICAL TRANSACTIONS OF THE ROYAL SOCIETY, 17TH-19TH CENTURIES
The Royal Society of London and the Philosophical Transactions (PTRS) have played an important role in the promotion of science and knowledge ever since their founding in the 1660s. Not only the British but also scholars and researchers from all over the world ‒ among whom many Italians ‒ have been interested in corresponding with the Society, becoming members and/or publishing on the Society’s journal. The PTRS thus became a focal point of cultural interaction between researchers from different countries. Moreover, by considering the journal in diachronic perspective we can see the gradual development of present-day scientific writing. My purpose is to investigate Italian contributions the Philosophical Transactions from a linguistic, historical and cultural perspective focusing on English and Italian relationships from the journal’s creation in 1665 up to the end of the 19th century. In this respect, the present piece of research focuses on a largely unexplored area in the history of Anglo-Italian socio-cultural relations, that is to say the scientific interactions between English and Italian researchers at the time when modern science was born and developed. The present study is a historical and critical linguistic analysis of PTRS articles written by Italians or based on Italian research and by analysing English and Italian relations through the papers and the epistolary exchanges of the scientists from the two countries. The aim from the linguistic perspective is to describe the features and development of Italian and Italian-research-inspired scientific writing in the Transactions; and ultimately, from the historical and cultural point of view, to provide a picture of Anglo-Italian relations in scientific context. The critical linguistic analysis of the primary sources here becomes functional to an objective analysis of cultural relations. It moreover adds to the existing research on the development of scientific writing by providing a study that is focused on a culturally-restricted group of papers and distinguishes between the sources of the writings. Comments and descriptions on editorial and translation practices will also be provided.
SCIENTIFIC CROSSCURRENTS BETWEEN ITALY AND ENGLAND: ITALIAN CONTRIBUTIONS TO THE PHILOSOPHICAL TRANSACTIONS OF THE ROYAL SOCIETY, 17TH-19TH CENTURIES / L. Berti ; tutor: G. Iamartino ; valutatori esterni: F. Henderson, M. D'Amore ; coordinatore del dottorato: M. V. Calvi. Dipartimento di Lingue e Letterature Straniere, 2020 May 05. 32. ciclo, Anno Accademico 2019. [10.13130/berti-lucia_phd2020-05-05].
CHARACTERISATION OF AUXIN RESPONSE FACTOR 5/MONOPTEROS POST-TRANSCRIPTIONAL AND POST-TRANSLATIONAL REGULATIONS DURING ARABIDOPSIS THALIANA DEVELOPMENT
Alternative splicing generates a MONOPTEROS isoform required for ovules developmentAuxin is one of the main designers of plant development and organs differentiation and its function relies on its differential accumulation with the generation of points of high and low hormone content. At cellular level, auxin signaling impacts on the transcriptomic landscape thought the modulation of the interplay between AUX/IAA repressor proteins and AUXIN RESPONSE FACTOR transcriptional factors. AUX/IAAs and ARFs compose a combinatory system in which AUX/IAAs binding to ARFs negatively regulates their ability to activate the transcription of downstream targets. Upon auxin sensing, AUX/IAAs stability is compromised thought their direct ubiquitination and degradation, allowing transcriptional activator ARFs to promote gene expression. Among transcriptional activator ARFs, ARF5/MONOPTEROS (MP) has been shown to be a key integrator of auxin signaling for many developmental events, thought the activation of the transcription of target genes above auxin threshold concentration. We show that in ovules, MP is expressed and functional in domains of low auxin content. We described that MP post-transcriptional regulations are involved in this process thought the generation of a biologically functional mRNA splicing variant, whose translation generates a protein isoform lacking the AUX/IAA interaction domain. Indeed, the alternative MPint11 isoform was able to partially complement mp mutants defects and ovule developmental process. Interestingly, also the canonical MP protein results partially functional for complementation of ovules formation, with a similar behavior to the one observed for MPint11. Our results suggest that MP functions relies on the combined activity of both its isoforms with the evidence that, during ovules development, MP domains involved in AUX/IAAs binding are not strictly required. These findings describe a novel scenario in which, not only MP is able to work uncoupled from the classical auxin signaling model, but also in which post-transcriptional regulation controls the formation of a functional isoform required for correct ovules development.
CHARACTERISATION OF AUXIN RESPONSE FACTOR 5/MONOPTEROS POST-TRANSCRIPTIONAL AND POST-TRANSLATIONAL REGULATIONS DURING ARABIDOPSIS THALIANA DEVELOPMENT / A. Cavalleri ; tutor: L.Colombo ; supervisore: M.Cucinotta ; phd director: M. Kater. Dipartimento di Bioscienze, 2021 Apr 14. 33. ciclo, Anno Accademico 2020.
NATURE AND GREEN CHEMISTRY AS KEY FACTORS FOR THE SYNTHESIS OF HIGH-QUALITY APIS FOR THE PHARMACEUTICAL INDUSTRY
Climate change has increased the need of an innovative approach to the application of chemistry in all fields with the aim to reduce the environmental impact of pharmaceutical industrial processes. From this perspective, green syntheses are more convenient for pharmaceutical companies and global society due to economic, environmental and safety reasons. Moreover, climate impact of pharmaceutical processes can be reduced by a clever exploitation of nature. Indeed, biomasses are an important source of active pharmaceutical ingredients (APIs) and complex scaffolds that can be used as starting materials or intermediates for the preparations of drugs, reducing hugely the number of synthetic steps. Obviously, a shorter synthesis of an API leads to a decrease of its price, becoming affordable to a larger number of people.In addition to small molecules, peptides are an important class of APIs, and their use has strongly increased during the years. The main drawback of peptides composed by coded amino acids is their poor pharmacokinetic profile due to their proteolytic susceptibility. A trick to improve this property is to insert a non-coded amino acid in the sequence. Taking inspiration from Nature, many researchers investigated the synthesis of new and exotic non-coded amino acids for different purposes and applications both as small molecules or when inserted in peptidomimetics. Nature and Green chemistry were the key factors of my PhD research, which aimed to develop more convenient syntheses of APIs. In particular, the goals of my research are divided into two main topics, involving:1. the synthesis or semi-synthesis of molecules from Nature inspiration, following the guidelines of Indena S.p.A., the partner who co-funded my PhD grant, together with PON REACT-EU financing Program;2. the synthesis of non-coded amino acids, coherently with the research field characterizing the research group of the University of Milan, in which I worked in these three years.
NATURE AND GREEN CHEMISTRY AS KEY FACTORS FOR THE SYNTHESIS OF HIGH-QUALITY APIS FOR THE PHARMACEUTICAL INDUSTRY / F. Anastasi ; tutor: M. L. Gelmi; coordinator: G. Vistoli. Dipartimento di Scienze Farmaceutiche, 2025 May 13. 37. ciclo, Anno Accademico 2024/2025.
EPIDEMIOLOGICAL AND FUNCTIONAL APPROACHES TO EVALUATE THE EFFECTS OF PARTICULATE MATTER EXPOSURE IN INFANTS WITH VIRAL BRONCHIOLITIS
According to the World Health Organization (WHO), environmental air pollution is currently among the leading causes of death and disease globally. Particulate matter (PM) exposure has been linked to the exacerbation of acute respiratory diseases, especially in the most susceptible population’s subgroups, such as infants and chil\dren. Acute bronchiolitis, a common clinical condition characterized by inflammation and obstruction of bronchioles, is generally caused by a viral infection of the lower airways and is estimated to be the leading cause of hospitalization of infants worldwide. At present, the severity of the disease is only partially explained by current known predisposing risk factors but recent evidence suggests that short-term exposure to PM causing airway inflammation, might result in increased susceptibility to viral infections and bronchiolitis severity.Due to its anatomical location, the upper respiratory tract (URT) is persistently exposed to PM, which affects the airway microbiome: recent evidence has linked the effects of PM10 and PM2.5 to microbiota structure and composition. In this context, innate immunity plays a fundamental role in the first response to both exogenous agents and commensal/pathobionts components of microbiota. In particular, bacterial-derived Extracellular vesicles (bEVs) are emerging as essential interregnum messengers that could play a crucial role in communication with mammalian cells, by modulating the host immune system.The main aim of the work is to investigate how PM exposure might modulate the relation that occurs between the severity of acute bronchiolitis, the respiratory microbiota, and the host immune response mediated by bEVs. First, we showed that the exposure to PM2.5 from the Milan metropolitan area impaired the innate immunity response following an acute infectious stimulus in a zebrafish (Danio rerio) embryo model. In particular, PM induced a pro-inflammatory effect, which leads to a dampened leukocytic response to a subsequent inflammatory stimulus. In the second part of the thesis, we investigated the relationship between short-term exposure to PM (PM2.5 and PM10) and the severity of viral bronchiolitis through a population study involving infants with bronchiolitis (with and without Respiratory Syncytial Virus, RSV, infection) aged less than one year living in the area of Milan. A positive association was observed between the severity of the disease and PM2.5 and PM10 exposure levels in the third week before the bronchiolitis peak. Moreover, we performed a metataxonomic analysis, comparing the composition of the bacterial portion of the nasal microbiome (bNM) of infants with bronchiolitis and matched it to healthy controls, to unveil a plausible interplay of the URT microbiota in this framework. We observed a respiratory dysbiosis in bNM of infants with bronchiolitis. Furthermore, Haemophilus influenzae (Hi) was predicted as the most represented species and also positively modulated by PM exposure during bronchiolitis. Last, the immunomodulatory potential of Hi-deriving bEVs was investigated using the zebrafish embryo. With this model, we showed that Hi-bEVs promote a pro-inflammatory response by altering leukocyte trafficking in particular in the neutrophil population.Overall, our data support the role of PM exposure in influencing the interplay between respiratory viruses, such as RSV, and specific microbiota components of the URT, such as Hi, in modulating the host immune response, potentially determining the pathogenesis and severity of acute bronchiolitis.
EPIDEMIOLOGICAL AND FUNCTIONAL APPROACHES TO EVALUATE THE EFFECTS OF PARTICULATE MATTER EXPOSURE IN INFANTS WITH VIRAL BRONCHIOLITIS / M. Cafora ; tutor: V. Bollati ; coordinatore: F. Ficetola. Dipartimento di Scienze Cliniche e di Comunità, 2023 Jul 11. 35. ciclo, Anno Accademico 2022.
CHROMOSOME-LEVEL DE NOVO GENOME ASSEMBLIES OF AVIAN SPECIES AND THEIR RELEVANCE FOR COMPARATIVE GENOMICS, PANGENOMICS, POPULATION GENOMICS AND SPECIES CONSERVATION.
Life on Earth is currently experiencing the sixth mass extinction. A major loss of species is being caused by anthropogenic habitat destruction, illegal wildlife trade, overfishing, massive fossil fuel consumption and the consequent climate changes, leading to a progressive collapse of biodiversity. The establishment of conservation initiatives is now more crucial than ever and the genetic management of threatened species is currently gaining critical significance. Previous genetic studies have been limited by the lack of complete reference genomes, thus having to focus on a limited number of genes or incomplete genomic sequencing data. In several cases, this led to wrong or incomplete assumptions and to conservation decisions which did not carry the desired effects due to incorrect evaluations. The availability of high-throughput sequencing technologies and the development of advanced computational methods have recently allowed the generation of cost-effective chromosome-level reference genomes. In the last decades, tremendous efforts were made to generate the most complete human genome. However, additional reference genomes spanning the entire tree of life are a necessary foundation for the study of biology in the 21st century, which was made possible with the establishment of large sequencing consortia. This thesis work represents a contribution to the collective effort of describing and preserving our planet's genetic diversity, being part of international consortia such as the Vertebrate Genomes Project (VGP), Darwin Tree of Life (DToL) and European Reference Genome Atlas (ERGA). Here I report new methods to assemble highly contiguous reference genomes for vertebrate species in the context of the VGP and their relevance in deciphering the biology of a species, its evolution, its intrinsic variability and in planning conservation actions, providing a comprehensive collection of genomic markers. Using the VGP pipelines, I have assembled 6 chromosome-level bird species: barn swallow (Hirundo rustica), lesser kestrel (Falco naumanni), American flamingo (Phoenicopterus ruber), common yellowthroat (Geothlypis trichas), rifleman (Acanthisitta chloris) and red-crested turaco (Tauraco erythrolophus), which fulfilled and exceeded the standard VGP metrics for genome assembly. Moreover, I contributed to the generation of the European nightjar (Caprimulgus europaeus) reference genome for the DToL. The genomes I generated will be exploited in the future to get insights into the biology of these species, but also for comparative genomics analyses using all available VGP-quality species. In particular, the lesser kestrel reference genome will be a fundamental resource for a future study that involves the assessment of how this species coped with climatic fluctuations in the past and how it is expected to cope with them in the future under the current scenario of climate changes. Moreover, the European nightjar genome will help to deepen the knowledge on the biology of this cryptic and2elusive bird, also boosting the sequencing of other members of the Caprimulgidae family to reconstruct a comprehensive phylogeny. During my PhD, I particularly focused on barn swallow (Hirundo rustica) genomics, an iconic migratory passerine bird with a long-standing association with humans. Using the VGP-quality assembly, I performed comparative genomics, population genomics and pangenomics analyses. Comparative genomics work was carried out with the reference-free aligner Cactus, with which I have aligned the barn swallow reference with other chromosome-level bird species. Using the alignment, I performed a positive and negative selection analysis across the genome to find candidate genes under selection important for barn swallow biology. For population genomics analyses, we aligned all publicly available data for the barn swallow subspecies and populations to the reference genome and performed a Linkage Disequilibrium scan. Comparative and populatio
CHROMOSOME-LEVEL DE NOVO GENOME ASSEMBLIES OF AVIAN SPECIES AND THEIR RELEVANCE FOR COMPARATIVE GENOMICS, PANGENOMICS, POPULATION GENOMICS AND SPECIES CONSERVATION / S. Secomandi ; Tutor: L. Gianfranceschi ; Co-tutor: R. Ambrosini, G. Formenti ; Coordinatore dottorato: F. Ficetola. Università degli Studi di Milano, 2022 Jun 20. 34. ciclo, Anno Accademico 2021.
TRANSCRIPTOMIC AND PROTEOMIC PROFILING OF PRECLINICAL SYSTEMIC SCLEROSIS
Background: Systemic sclerosis (SSc) is a rare systemic autoimmune disease with a high morbidity and mortality. In the preclinical phase of the disease, patients present exclusively Raynaud phenomenon (RP) with a nailfold video-capillaroscopy positive for scleroderma pattern and/or the presence of specific autoantibodies. Research investigations on preclinical systemic sclerosis (Pre-SSc) patients are rare due to the exceptionality and the difficulty to intercept this specific subgroup of subjects that in five years will progress into a definite SSc diagnosis in about 50% of cases. Objectives: to perform a comprehensive investigation of preclinical SSc at gene expression, proteomic and clinical level with the aim to identify specific early-stage disease biomarkers as well as progression biomarkers. Methods: gene expression analysis through RNAseq technique was performed on whole blood samples of 35 Pre-SSc and 16 matched healthy controls (HC) collected at baseline and after four years. Gene expression module changes analysis was performed comparing evolving Pre-SSc, stable Pre-SSc and HC at baseline and follow-up. A proteomic analysis through SOMAscan technology was assessed on serum at baseline of a subgroup of 16 Pre-SSc (8 evolving Pre-SSc vs 8 stable Pre-SSc) and 8 HC selected on a clinical basis, matching for age, gender and autoantibody profile. Finally, proteins emerged from SOMAscan analaysis to be predictive of progression were validated on a validation cohort of 50 subjects with preclinical features whose serum aliquots were available at baseline. Results: out of 35 Pre-SSc, 15 (42.9%) progressed toward a definite SSc after four years. The presence of gastro-esophageal reflux and a shorter time of RP were associated with a shorter time of progression.RNA expression change analysis of evolving Pre-SSc vs stable Pre-SSc identified 73 genes with a corrected p value ≤0.05. At baseline, Pre-SSc had type I IFN gene expression modules (M 3.4, M 5.12, M 1.2) increased in comparison to HC. NK gene expression modules were decreased in evolving Pre-SSc over time. Out of 286 proteins assessed by SOMAscan, 10 proteins were able to predict at baseline progressors from not progressors. Evolving Pre-SSc showed increased levels of NKp30, Endostatin, bFGF, ECM1, FGF18, Fibronectin 1.3, PAFAH1B2, FABP and decreased levels of PHI and Ubiquitin1. High levels of endostatin and reduced serum levels of PAFAH1B2 were confirmed with ELISAs in the validation cohort to correlate with a shorter time to progression. Conclusion: a type I IFN signature distinguished preclinical SSc from HC and a reduced NK signature was associated to SSc progression. Proteins linked to pathways of fibrosis, extracellular matrix organization, positive regulation of cell proliferation, angiogenesis, signal transduction were discovered to predict disease progression. Moreover, endostatin emerged as a biomarker worthy of future mechanistic investigations.
TRANSCRIPTOMIC AND PROTEOMIC PROFILING OF PRECLINICAL SYSTEMIC SCLEROSIS / C. Bellocchi ; tutor: N. Montano ; supervisore: L. Beretta ; coordinatori: E. Berti, M. Del Fabbro. Dipartimento di Scienze Cliniche e di Comunità, 2021 Dec 14. 34. ciclo, Anno Accademico 2021.
TESTI SCOLASTICI DELLA LOMBARDIA DEL XIV E XV SECOLO.TENDENZE KOINEIZZANTI E INDIZI DI GRAMMATIZZAZIONE DEL VOLGARE LOMBARDO
This work aims to study the vernacular language used in school texts intended for the teaching of Latin in Lombardy in the 14th and 15th centuries. Starting in the 14th century, bilingual textbooks in Latin and the vernacular began to appear in schools, in which the vernacular was used for the (practical) purpose of making the Latin text accessible. The research aims, on the one hand, to provide an analysis of the vernacular language used in these texts to describe certain tendencies towards the koineisation of Old Lombard, and, on the other, to investigate the relationship between Latin and vernacular in school texts and its evolution. The reflection on Latin grammar led teachers and students of the time to confront themselves with linguistic problems that were also applicable to their mother tongue. An attempt was made to ascertain the extent to which these reflections influenced the process of grammatisation and normativisation of Old Lombard at the time.
TESTI SCOLASTICI DELLA LOMBARDIA DEL XIV E XV SECOLO.TENDENZE KOINEIZZANTI E INDIZI DI GRAMMATIZZAZIONE DEL VOLGARE LOMBARDO / A. Soma ; cotutor: G. Polimeni ; tutor: R. Wilhelm ; coordinatore: F. Slavazzi. Dipartimento di Studi Letterari, Filologici e Linguistici, 2024 Sep 30. 36. ciclo, Anno Accademico 2023.
The present study aimed to assess udder health by estimating the prevalence of subclinical mastitis (SCM) and identifying intramammary infection (IMI) causing pathogens. Furthermore, milk quality was evaluated by measuring bulk milk somatic cell count (BMSCC) on buffalo farms and total bacterial count (TBC), total non-aureus staphylococci count (TNAS), total Staphylococcus (S.) aureus count (TSA), total Enterobacteriaceae count (TEC), and the presence of zoonotic pathogens along the water buffalo milk chain in Bangladesh. A total of 248 water buffalo farms from nine buffalo concentration districts, comprising 51 free-range, 106 semi-free-range, 33 households, 54 semi-intensive, and four intensive buffalo farms, were visited to collect individual quarter milk to diagnose SCM using CMT; BMSCC was also measured on-farms. One hundred twenty-two farm bulk milk samples, 109 middlemen milk samples, 111 milk collection centers, and 35 milk product samples were collected and assessed for TBC, TNAS, TSA, and TEC. A subset of these samples, comprising farm bulk milk (n = 34), the middleman (n = 37), the milk collection center (n = 37), and milk products from shops (n = 35), were analyzed using real-time PCR to estimate the prevalence of seven important zoonotic pathogens: S. aureus, Escherichia (E.) coli, Shiga toxin-producing E. coli O157:H7, Campylobacter (C.) jejuni, Listeria (L.) monocytogenes, Salmonella (S.) enterica, and Yersinia (Y.) enterocolitica. Quarter, buffalo, farm level, and milk value chain-related data was collected using questionnaires and observations. The overall SCM prevalence was high at the quarter level (27.9 %) and the buffalo level (51.5 %). Non-aureus staphylococci were the predominant IMI-associated bacterial species. The proportion of non-aureus staphylococci or Mammaliicoccus sciuri (NASM), S. aureus, and other bacterial species identified in the buffalo quarter samples varied between buffalo farms. An intensive buffalo-rearing system, teats on the left, funnel-shaped teats, udder asymmetry, single milker, and a lack of quarantine facility were all associated with poor buffalo udder health. The geometric mean of BMSCC was 217,000 cells per mL of milk, which was low on average, but some farms could improve substantially. Farm-hygiene-related risk factors for IMI were poor milking hygiene and cleanliness of the hind quarters. The likelihood of IMI from any pathogen and NASM were higher in buffalo herds with poor milking hygiene. Poor cleanliness of the hind quarters increased the chances of IMI caused by any pathogen, NASM, and S. aureus. Farm management-related risk factors for IMI were the milking frequency and buffalo source. Farms with buffalo purchased from another herd were associated with IMI from any pathogen. Udder symmetry and body condition score (BCS) were risk factors related to buffalo breeding and nutrition. Asymmetrical udders were associated with IMI caused by any bacteria and with IMI caused by S. aureus. Poor BCS showed higher odds of IMI from any pathogen and by NASM. Progressively increasing TBC, TNAS, and TEC levels were observed along the milk value chain (P < 0.05). High TBC was associated with the winter season, cleaning the milk containers using pond water, a mix of buffalo and cow milk, and a poor cleanliness score for the milk containers. Farms located in coastal areas were associated with high levels of TNAS. Zoonotic pathogens' prevalence increased over the water buffalo milk value chain. Three classical enterotoxin-encoded genes for E. coli O157:H7 were tested, eae, stx1, and stx2, of which the stx2 genotype was the most prevalent in milk products (74 %). L. monocytogenes and Y. enterocolitica were more prevalent in the farm bulk milk (65-79 %) than in the later milk value chain nodes. The prevalence of S. enterica was somewhat low (0 - 14 %) in all the milk chain nodes, and all the samples were negative for C. jejuni. Using plastic containers and dirty clothes to clean milk
WATER BUFFALO MILK CHAIN IN BANGLADESH / S. Singha ; tutor: F. CECILIANI ; supervisors: F. CECILIANI, Howlader, Md. Matiar Rahman, Persson, Ylva, Koop, Gerrit; Rahman, Md. Mizanur ; phd coordinator: F. Ceciliani. Dipartimento di Medicina Veterinaria e Scienze Animali, 2023 Oct 23. 35. ciclo, Anno Accademico 2022.
THE GENDERED IMPACT OF THE PANDEMIC: EMPLOYMENT, INCOME AND UNPAID DOMESTIC WORK DURING COVID - 19 PANDEMIC IN EUROPE.
The COVID-19 pandemic brought sudden and profound disruptions to daily life across the globe, reshaping how people work, care, and live. Compared to recent global crisis where the initial impact on employment was more severe for men, the COVID-19 pandemic crisis had a significant initial impact on sectors with higher share of women. The abrupt pandemic driven changes have raised the alarms on the potential detrimental impact this crisis might have on three crucial areas of already persistent gender inequalities on global level—unpaid domestic and care work, employment and earnings. Previous research on the gendered impact of the pandemic has focused mostly on the initial period of the pandemic and offers limited comparative perspective with respect to the contextual, individual, and household level characteristics. Building on the current literature, this thesis aims to explore how the COVID-19 pandemic has affected gender inequalities in employment, income, and unpaid domestic work and childcare across different European contexts, and which are the main underlying mechanisms on individual, meso and macro level that shape the gendered outcomes in times of global pandemic crisis. The first study examines how the division of unpaid domestic work and childcare in couples evolved during the initial shock of the pandemic and the early recovery period in two distinct European welfare and gender regimes, conservative (Germany) and liberal (the UK). Additionally, it explores the role of couples’ employment status in shaping the distribution of unpaid labour during the extended impact of the pandemic. The results show shift towards more equal division of unpaid housework and childcare in the lockdown and post-lockdown period within couples in the UK. In Germany, shift towards more equal division is only evident for childcare, while division of housework remains without notable shifts compared to the pre-pandemic period. Despite the certain shifts towards less inequality, however, the unequal division of housework and childcare remained significant, particularly for Germany, indicating that the unpaid labour in households (particularly housework) during the pandemic crisis was still mostly considered woman’s responsibility, although the context and the individual and couples’ characteristics, played an important role. The second study investigates the gendered employment outcomes during the pandemic across European countries. It examines the underlying factors on micro-level (i.e. education, employment sector and type of contract, and presence of children in household) and macro-level (i.e. the strictness of the lock-down measures, and countries’ gender inequality levels) which shape the differences in the negative employment outcomes during the pandemic for women and men across Europe. Moreover, it explores the interaction between the micro-level characteristics and macro-level context, indicating how individual-level characteristics are of different importance in different country contexts in times of exogenous shock and crisis. The findings reveal that on individual level, better education, employment sector and unlimited job contract, served as protective factors against the negative employment outcomes during the pandemic, offering consistent protection for both women and men. On the contextual level, the strictness of the pandemic containment measures was particularly relevant for men’s employment outcomes, while the country’s gender inequality levels affected both women and men. The interaction between the individual and contextual characteristics indicates that countries with higher level of gender equality provide more favourable and protective context against negative employment outcomes for women in times of external global economic shocks as the COVID-19 pandemic. Lastly, the third study shifts the focus on the relative income changes during in the first year of the COVID-19
THE GENDERED IMPACT OF THE PANDEMIC: EMPLOYMENT, INCOME AND UNPAID DOMESTIC WORK DURING COVID - 19 PANDEMIC IN EUROPE / M. Bashevska ; tutor: G. M. Dotti Sani ; coordinator: P. A. Rebughini. - Department of Social and Political Sciences, University of Milan.. Università degli Studi di Milano, 2025 Jul 02. 36. ciclo, Anno Accademico 2023/2024.
LIRE DES ROMANS CHEZ EINAUDI: LES CAS DE NATALIA GINZBURG (1944-1964). LE RAPPORTS DE LECTURE AU PRISME DES HUMANITES NUMERIQUES
The objective of this thesis is to study the editorial activity of Natalia Ginzburg at Einaudi publishing house, in particular as an editorial reader of contemporary narrative works, from her arrival to the temporary interruption of the collaboration (1944-1964). We analyse her contribution to decision-making processes, to the construction of the identity and of the catalogue of the publishing house and, more generally, to the definition of the literary culture of her time. Our objective is, moreover, to define the literary models proposed by Natalia Ginzburg to Einaudi’s readers, the way in which her work was part of the intellectual laboratory of Einaudi and in which her idea of literature harmonized with market issues, the relationship that existed between her opinions on the economic and literary viability of a work and the historical-political context, the method and criteria she used to evaluate a book, the relationship that existed between her choices as a publisher and her poetics as an author.More specifically, our thesis aims to analyse the reading reports concerning contemporary narrative works, which represent a fundamental step in the decision-making process that leads to the publication of a work and therefore to the construction of the catalogue of a publishing house. Through these documents, the books to be printed were selected. Reading reports therefore embodied cultural models that need to be interpreted according to a specific historical and cultural context. Concerning the processing of these documents, a database for the inventory of sources has been designed with IT support. This database makes it possible to efficiently account for sources by gathering the fundamental information for the description of the corpus in a structured way. The documents were indeed transcribed and encoded according to a model developed in XML TEI, able to highlight the different types of documents (letters, structured reading reports, minutes, etc.), their structure and the existing relationships between them.The modelling activity had an important methodological and heuristic impact to our work. The tool that was created shows the complexity of the corpus of sources, by highlighting paths that are not immediately evident in a linear analysis of documents, by allowing query and by enhancing their intertextuality and their multiautoriality.The examination of her reading reports, mostly unpublished, provides the basis for a critical reflection on Ginzburg’s editorial activity, a reflection which is rooted in the belief that editorial writing represents a complex critical genre that deserves to be investigated by specific methods.
LIRE DES ROMANS CHEZ EINAUDI: LES CAS DE NATALIA GINZBURG (1944-1964). LE RAPPORTS DE LECTURE AU PRISME DES HUMANITES NUMERIQUES / L. Antonietti ; tutor: E. PIERAZZO ; cotutor: A. CADIOLI. Università degli Studi di Milano, 2022 Mar 25. 32. ciclo, Anno Accademico 2019.
A MULTI-OMICS APPROACH TO DISSECT ABERRANT EPIGENETIC MECHANISMS IN TRIPLE NEGATIVE BREAST CANCER
In recent years, experimental evidence has shown that epigenetic changes including DNA methylation and histone post-translational modifications (PTMs) play a role in cancer development and progression. Histone PTMs have been largely studied in the context of cancer and have been described as hallmarks of cancer and markers of diagnosis and prognosis. In addition, the histone modifying enzymes involved in the recognition, deposition, and removal of histone PTMs are often altered and overexpressed in cancer. Therefore, the identification of these enzymes as potential targets for epigenetic therapies is promising for the development of specific treatments. In this project, we focused on the molecular characterization of epigenetic mechanisms in Triple Negative breast cancer (TNBC), which is the most aggressive type of breast cancer, characterized by a high proliferation rate, and associated with poorer patient outcomes compared to other breast cancer subtypes, particularly Luminal A subtype. Because of the absence of specific molecular markers, there are no targeted therapies available for TNBC and only few treatment options are in use, including standard chemotherapy, surgery, and radiation therapy. Therefore, there is an urgent need to identify specific markers for this type of tumor as well as novel potential targets. Preliminary data obtained by our group, by mass spectrometry (MS), highlighted an epigenetic signature distinguishing the TNBC subtype from the other breast cancer subtypes. We confirmed the same pattern in an additional set of samples and we selected three of the identified changes (H3K4me2, H3K9me3, and H4K20me3) for a follow-up mechanistic investigation, using two parallel approaches. We first performed a multi-OMICs analysis to investigate the genomic distribution of H3K4me2 in TNBCs and its effects at the expression level. By integration of ChIP-seq and RNA-seq data from a set of fresh frozen clinical samples we found that the localization of H3K4me2 at promoters and super-enhancer regions results in higher levels of expression and seems to drive expression of genes associated to the TNBC phenotype. By the integration of data from clinical samples and the model breast cancer cell line MDA-MB-231 we identified a set of genes that are candidate TN-specific genes whose expression is regulated by the unique presence of H3K4me2 in their cis-regulatory regions. We then took advantage of a CRISPRi epigenetic tool to genetically manipulate the expression of these genes through the LSD1-mediated removal of H3K4me2 at these regulatory regions. The result obtained revealed a direct correlation between the presence of H3K4me2 at promoters and gene expression. Then, we investigated the possible upstream causes of histone PTM deposition focusing on H3K9me3 modification, which we found to be overexpressed in TN versus Luminal A tumors. By correlating the levels of histone PTMs in TNBC samples with the expression levels of the histone modifying enzymes by RNA-seq analysis, we found a strong correlation between the increase in H3K9me3 levels and the over-expression of the H3K9-selective methyltransferases SETDB1, SUV39H1, and SUV39H2 in TN subtype. Upon pharmacological and genetic manipulation of these methyltransferases, we identified a strong effect at phenotypic level. In addition, the knockdown of SUV39H1 and SUV39H2, but not SETDB1, revealed a decrease in H3K9me3 levels paralleled by an increase in H4K20me3 levels, suggesting a potential mechanism of crosstalk. Hence, we hypothesized that upon depletion of SUV39H1 and SUV39H2 a re-localization of both H3K9me3 and H4K20me3 may occur. To assess this hypothesis, we used immunofluorescence to analyze the localization of H3K9me3 and H4K20me3 on mitotic chromosomes and correlated these data with ChIP-seq analysis of these two epigenetic marks in a set of Luminal A and TN cell lines. At basal condition, we observed a more pronounced localization of these histo
A MULTI-OMICS APPROACH TO DISSECT ABERRANT EPIGENETIC MECHANISMS IN TRIPLE NEGATIVE BREAST CANCER / G. Robusti ; internal advisor: S. Minucci ; phd coordinator: S. Minucci ; tutor: T. Bonaldi. Dipartimento di Oncologia ed Emato-Oncologia, 2023 Dec 12. 35. ciclo, Anno Accademico 2023.
LONGITUDINAL NUTRITIONAL INTAKES IN ITALIAN PREGNANT WOMEN IN COMPARISON WITH NATIONAL NUTRITIONAL GUIDELINES AND PLACENTAL ANTIOXIDANT DEFENSES AND AUTOPHAGY-RELATED GENES IN MATERNAL OBESITY AND GESTATIONAL DIABETES MELLITUS
Nutritional quality during pregnancy is crucial for the health status of mother and child. Maternal obesity and gestational diabetes mellitus (GDM) are increasing worldwide, representing principal risk factors for both mother and child short/long-term outcomes. Oxidative stress, lipotoxicity and altered autophagy have already been reported in obesity, but few studies have focused on obese pregnant women with GDM.The aim of this Doctoral Thesis is to:- evaluate the adherence to pregnancy nutritional recommendations during the three pregnancy trimesters in Italy in pre-gestational Normal Weight and Over Weight women;- evaluate antioxidant and macro/chaperone-mediated autophagy (CMA)-related gene expression in obese and GDM placentasData presented in the first study derived from the analyses of 176 women (NW=133; OW=43) with a healthy singleton pregnancy, enrolled within 13+6 weeks of gestation. Dietary intake was assessed with a Food Frequency Questionnaire (FFQ) in each trimester.Gestational weight gain (GWG) did not differ between subgroups. Recommendations for caloric intake were never reached from both subgroups in each trimester. Proteins intake was higher than recommendations in first and second trimester for both subgroups. Sugars percentage was always increased in both subgroups while dietary fiber intake decreased in OW. Polyunsaturated fatty acids, calcium, iron and folic acid requirements were never satisfied in both subgroups, while sodium intake was always greater than suggested. These results indicate that both NW and OW women in Italy do not adhere to nutritional recommendations during pregnancy. Inadequacies in micronutrients intake were also observed. Nutrition in pregnancy needs adequate survey and educational intervention as well as supplementation when indicated.In the second study a total of 47 women with singleton pregnancies delivered by elective cesarean section were enrolled: 16 normal weight (NW), 18 obese with no comorbidities (OB GDM(–)), 13 obese with GDM (OB GDM(+)). Placental gene expression was assessed by real-time PCR.Antioxidant gene expression (CAT, GPX1, GSS) decreased, the pro-autophagic ULK1 gene increased and the chaperone-mediated autophagy regulator PHLPP1 decreased in OB GDM(–) vs. NW. On the other hand, PHLPP1 expression increased in OB GDM(+) vs. OB GDM(–). Sexual dimorphism for both antioxidant and CMA-related gene expressions was found. These preliminary results can pave the way for further analyses aimed at elucidating the placental autophagy role in metabolic pregnancy disorders and its potential targetability for the treatment of diabetes outcomes.
LONGITUDINAL NUTRITIONAL INTAKES IN ITALIAN PREGNANT WOMEN IN COMPARISON WITH NATIONAL NUTRITIONAL GUIDELINES AND PLACENTAL ANTIOXIDANT DEFENSES AND AUTOPHAGY-RELATED GENES IN MATERNAL OBESITY AND GESTATIONAL DIABETES MELLITUS / F. Lisso ; tutor: I. Cetin ; co-tutor: C. Mandò; co-supervisori: C. Novielli, G.M. Anelli ; coordinator: L. Pinotti. Dipartimento di Scienze Biomediche e Cliniche L. Sacco, 2022 Apr 28. 33. ciclo, Anno Accademico 2020.
SEARCH FOR ELECTROWEAK PRODUCTION OF SUPERSYMMETRIC PARTICLES IN COMPRESSED MASS SPECTRA WITH THE ATLAS DETECTOR AT THE LHC
In this thesis, two analyses searching for the production of supersymmetric particles through the electroweak interaction are presented: the charginosearch, targeting the pair production of charginos decaying into W bosons and neutralinos, and the displaced track search, looking for mildly displaced charged tracks arising from the decays of higgsinos into pions. These searches target compressed phase spaces, where the mass difference between the next-to-lightest and lightest supersymmetric particle is relatively small. The searches use proton-proton collision data collected at a centre-of-mass energy of 13 TeV with the ATLAS detector at the LHC.In the chargino search, the targeted difference in mass between charginos and neutralinos is close to the mass of the W boson. In such phase space, the chargino pair production is kinematically similar to the WW background, which makes the chargino signal experimentally challenging to be discriminated from the WW background. A final state with two leptons from the leptonic decays of the W bosons, missing transverse momentum and no hadronic activity is considered, while advanced machine learning techniques are adopted to separate the supersymmetric signal from the backgrounds.Chargino masses up to about 140 GeV are excluded at 95% confidence level in the case of a mass splitting between chargino and neutralino down to about 100 GeV. The results supersede the previous ATLAS results in particularly interesting regions where the chargino pair production could have hidden behind the looking-alike WW background.In the displaced track search, the difference in mass between the produced supersymmetric particles and the lightest neutralinos goes down to 0.3 GeV.The experimental signature is one jet, missing transverse momentum, and a low momentum charged track with an origin displaced from the collision point, the last element being the first time it is used in a search of this kind at a hadron collider. The results show that the analysis has the sensitivity to exclude different signal hypotheses for higgsino masses up to 175 GeV if no excess is observed in data. For lower masses, the larger signal cross-section allows to achieve higher significance values for different mass splitting scenarios. All these signal hypotheses have not been probed by any existing analysis of LHC data.
SEARCH FOR ELECTROWEAK PRODUCTION OF SUPERSYMMETRIC PARTICLES IN COMPRESSED MASS SPECTRA WITH THE ATLAS DETECTOR AT THE LHC / E. Ballabene ; supervisor: T. Lari ; coordinator: M. Paris. Università degli Studi di Milano, 2022 Nov 17. 35. ciclo, Anno Accademico 2022.
ASTROPHYSICAL NUCLEAR REACTIONS ON NEON ISOTOPES AT LUNA
This thesis reports the direct measurements of the 22Ne(α,γ)26Mg and 20Ne(p,γ)21Nareactions at astrophysical energies of interest. The 22Ne(α,γ)26Mg reaction competes with the 22Ne(α,n)25Mg reaction which is the main source of neutrons for the s-process in low-mass Asymptotic Giant Branch and massive stars. At temperatures T < 300 MK where the (α,γ) channel becomes dominant, the rate of the 22Ne(α,γ)26Mg reaction is influenced by several resonances studied only indirectly. The first part of this thesis concerns the direct measurement of one of these resonances, Er = 334 keV, which so far was studied only indirectly leading to six orders of magnitude range of possible values for its resonance strength. The experiment has been performed at LUNA (Laboratory for Underground Nuclear Astrophysics) using the intense alpha beam of the LUNA 400 kV accelerator and a windowless gas target combined with a high-efficiency BGO detector. In the present study, an upper limit of 4.0·10−11 eV has been determined for the resonance strength. Taking into account these results, an up-dated 22Ne(α,γ)26Mg thermonuclear reaction rate was obtained and its role on the predicted 25Mg/26Mg ratio of a 5M⊙ AGBs was investigated. The data show a decrease by a factor of 15 of the intershell 25Mg/26Mg ratio.The 20Ne(p,γ)21Na is the slowest reaction of the NeNa cycle. It determines the velocity of the cycle and defines the final abundances of the isotopes synthesized in this cycle. The uncertainties on the NeNa cycle are affected by the 20Ne(p,γ)21Na reaction rate. The main goal of the second part of this thesis was the direct measurement of the Ecm = 366 keV resonance which dominates the total rate in the temperature range between 0.2 GK and 1 GK. The measurement has been performed at LUNA using the windowless gas target and two high-purity germanium detectors placed at different positions. This measurement allowed to reduce the uncertainty on the strengths of the 366 keV resonance from 18% to 7%. These results were used to update the 20Ne(p,γ)21Na reaction rate.
ASTROPHYSICAL NUCLEAR REACTIONS ON NEON ISOTOPES AT LUNA / E. Masha ; supervisor: A. Guglielmetti; co-supervisor: A. Caciolli; director of the school: M. Paris. Dipartimento di Fisica Aldo Pontremoli, 2022 Jan 21. 34. ciclo, Anno Accademico 2021. [10.13130/masha-eliana_phd2022-01-21].
AMIDYL RADICALS IN LIGHT PROMOTED REACTIONS
The present thesis work is devoted to the study of four different topics as depicted in Figure 1, the enantioselective addition of amidyl radicals in α position to aldehydes, (Figure 1A), the addition of N lactam radicals to nucleophiles such as heteroarenes, arenes and aldehydes (Figure 1B), the preliminary investigation of the enantioselective addition of N-acetyl radicals to olefins (Figure 1C) and the development of the enantioselective α-SCF3 addition to β-ketoesters (Figure 1D). Figure 1 Research outline. In particular, Chapter 1 provides an overall introduction to the main topics encountered in this PhD thesis: Chapter 1.1 deals with a general definition of photocatalysis, the main photocatalytic activation pathway and same selected literature example to compare light catalysed reactions with and non-light catalysed reactions. The nitrogen radicals are introduced and classified in Chapter 1.2 with an historical overview of the previous literature work, while Chapter 1.3 reports the main advantages to perform a photochemical transformation exploiting the flow technology.Chapter 2 is focused on the results obtained and it is divided in four main topics plus an appendix (illustrated in Figure 1): Chapter 2.1 deals with the enantioselective addition of amidyl radicals to aldehydes employing an imidazolidinone organocatalyst to promote the selectivity of the transformation. Moreover, we report also the development of the flow process for this transformation. Chapter 2.2 concerns the development and the study of a new class of amidyl radicals: the N lactam radicals. We are able to synthetize suitable nitrogen radical precursors and employ them in the photocatalytic addition of γ, δ, ε lactam radicals to heteroarenes, arenes and aldehydes. Moreover, we perform a wide reaction scope and a deepen mechanism investigation. After having deeply studied the employment of these new radicals, we selected a pharmaceutical target to applied the N lactam radicals and the results are reported in Chapter 2.3. A new photocatalytic synthesis of Levetiracetam, an antiepileptic drug, was developed. Chapter 2.4 reports the idea and the initial attempts done to develop the enantioselective addition of N-acetyl radicals to electronrich olefins. The experiments reported in the Appendix concerned the enantioselective addition of the enantioselective α-SCF3 addition to β-ketoesters were executed at the beginning of the PhD period before starting work on amidyl radicals in light promoted reactions. Chapter 3 provides all the experimental part of the PhD thesis including the synthetic procedures, 1H-NMR, 19F-NMR and 13C-NMR of the unknown compounds and the HPLC traces of the enantiomeric excess of the asymmetric transformations.
AMIDYL RADICALS IN LIGHT PROMOTED REACTIONS / M.f. Boselli ; tutor: A. Puglisi ; co-tutor: M. Benaglia ; coordinatore: D. Passarella. - Milano. Dipartimento di Chimica, 2024 Jun 14. 36. ciclo, Anno Accademico 2023/2024.
GAUSSIAN PROCESSES FOR 3D SHAPE MODELLING OF NOISY AND INCOMPLETE DATA. AN APPLICATION TO HUMAN EARS RECONSTRUCTION
Mathematical modelling of 3D shapes is a requirement in a vast and increasing number of applications,especially when it comes to the human shape. Despite being an active branch of research, with manyproposed solutions, some settings remain particularly challenging. An example of such an open problemis the modelling or reconstruction of human ears from 3D point clouds, the motivation behind this thesis.This specific problem proves to be a particularly challenging case due to the fine and non-linear detailsto be modelled, together with the extensive presence of missing data and outliers, originated from thedata retrieval procedure.Gaussian Processes were recently proposed as a suitable framework for the formulation of shapemodelling problems and have mostly been applied to the human face. We show that this is indeed themost promising setting for our particular kind of data, but it can not be trivially applied from existingsolutions. Therefore, we propose a fully unsupervised pipeline from raw 3D scans to a complete earmodel, able to cope with the specific challenges encountered.Nonetheless, the main bottleneck of the pipeline is found on the shape registration part, that hindersthe quality of the final results. We address this problem by incorporating the registration task entirelywithin the Gaussian Processes framework, unlike previous approaches and we provide a method forparameter estimation based on Variational Bayesian Inference. Not only do we achieve better registrationresults in the presence of large regions of missing data, but we provide a more unified way to deal withshape modelling. This is a step towards a more coherent approach and away from complex pipelines,merging different and often contradictory assumptions.
GAUSSIAN PROCESSES FOR 3D SHAPE MODELLING OF NOISY AND INCOMPLETE DATA. AN APPLICATION TO HUMAN EARS RECONSTRUCTION / F. Marreiros Malveiro Valdeira ; tutor: A. Micheletti, C. Soares, R.Ferreira ; coordinatore: D. Bambusi. Università degli Studi di Milano, 2022 Nov 25. 34. ciclo, Anno Accademico 2021.
OMICS APPROACHES FOR OMEGA-6/OMEGA-3 POLYUNSATURATED FATTY ACID RATIO IN PIGS
In general, meat quality is heavily influenced by fatty acid (FA) composition and intramuscular fat, which is mostly dependent on the dietary fat intake of pigs, but it is also affected by genetic variants. In this PhD thesis, we elucidated the effects of dietary lipids, in terms of omega-6 (n-6)/omega-3 (n-3) polyunsaturated fatty acid (PUFA) ratio, on porcine meat quality and the integration of OMICS technologies such as genomics, transcriptomics, and proteomics. With the aim of understanding the association between n-6/n-3 PUFA ratios, porcine transcriptome expression, and biological processes related to PUFA metabolism, we performed RNA-Seq and microRNA-Seq on longissimus dorsi muscle samples from 20 Iberian x Duroc pigs with extreme values for n-6/n-3 FA ratio and identified differentially expressed messenger RNAs (mRNAs) and microRNAs (miRNAs). Our findings highlighted mRNA genes, miRNAs and enriched pathways that were related to lipid metabolism, cell growth and inflammation, according to differences in muscle n-6/n-3 PUFA ratio. Relevant miRNA-to-mRNA regulatory networks were also uncovered (i.e., mir-15b to ARRDC3; mir-7142-3p to METTL21C), and linked to lipolysis, obesity, myogenesis, and protein catabolism. We also determined on how sow diets with high or low n-6/n-3 PUFA ratios of 13:1 (SOY) and 4:1 (LIN), influenced the reproductive performance of sows, growth performance of pre-weaned piglets, and fat deposition, including proteins and protein-coding genes using proteomics analysis. In sows, LIN decreased dead-born piglets and pre-weaning mortality. Although piglet weight and weight gain varied in between n-6/n-3 PUFA ratios, weaning weight and overall weight gain were not affected by dietary interventions. Proteomic analysis revealed 4 and 11 overabundant proteins in muscle and adipose tissues, respectively, in SOY compared to LIN. Additionally, the pro-inflammatory role of n-6 PUFAs may be associated to the observed overabundance of haptoglobin, an acute-phase protein, and the stimulation of protein-coding genes and proteins relevant to the innate immune response and acute inflammatory response. Using the same population, we applied the use of exon-intron split analysis (EISA) approach to account for post-transcriptional changes, driven by putative functional miRNA repression, in response to extreme values of n-6/n-3 PUFA ratio in longissimus dorsi muscle in male and female piglets. EISA revealed 11 and 97 mRNA genes within the top 5% negative PTc scores with at least 1.5-fold exonic region reduction in SOY-male (SOY-M) and LIN-male (LIN-M) piglets, respectively. Furthermore, the detected upregulated miRNAs harbored binding sites on 72.73% (SOY-M) and 61.86% (LIN-M) of these post-transcriptionally downregulated genes. Moreover, the overall PTc signals in females were not as strong and clear as those from the males. In addition, we discovered genes that are predominantly associated in the modulation of immune responses and lipid-related metabolism, which may be related to the pro- and anti-inflammatory properties of n-6 and n-3 PUFAs, respectively. Our use of EISA enabled us to identify regulatory networks that complemented traditional analyses of differential expression, providing us a more comprehensive understanding of muscle metabolic changes in response to PUFA concentration.
OMICS APPROACHES FOR OMEGA-6/OMEGA-3 POLYUNSATURATED FATTY ACID RATIO IN PIGS / Y.j. Manaig ; tutor: A. Sanchez, Folch, JM, G. Savoini; coordinatore: F. Ceciliani. Università degli Studi di Milano, 2022 Dec 21. 34. ciclo, Anno Accademico 2021.
FOOD SECURITY: INNOVATIVE PLANT REPRODUCTIVE STRATEGY IN A CHANGING ENVIRONMENTAL CONDITION
With the climate changing and the population increasing, the global food security issue is currently one of our main concerns. In this framework, the development of advanced plant breeding strategies is essential to ensure population feeding. Apomixis is an asexual reproductive mechanism in which the seeds are produced clonally. In apomictic plants, the female meiosis is altered or bypassed thus generating unreduced spores and consequently gametes that develop in embryos without fertilization. The advantage of this reproduction type is the fixation of highly resistant and productive varieties through generations, without segregation of the traits. However, the introduction of apomixis in crop plants is challenging, because its regulation is still largely unknown.During my PhD, I have characterized this process at the cytological and molecular level, highlighting the main differences with sexual reproduction especially during the first meiotic division. Furthermore, I have used different approaches to unveil the genes potentially involved in apomixis regulation in the natural apomictic species Taraxacum officinale – the common dandelion. I have contributed to characterize the molecular function of VACUOLAR PROTEIN SORTING 13 (VPS13), one putative regulator required for the formation of unreduced and clonal female gametes in dandelion, and ovule development in the model sexual species Arabidopsis thaliana, assessing its downstream pathways.Due to the lack of Taraxacum vps13 loss of function mutants, to understand the mechanism of action of VPS13 I have characterized in detail the Arabidopsis vps13 mutant. By studying ovule development, I have demonstrated that this gene is necessary for the correct specification of the female germline and the progression of megasporogenesis. In particular, I have established VPS13's role in the small RNA pathway and its interaction with meiotic proteins. All in all, my work has contributed to the understanding of the molecular regulation of sexual and apomictic reproduction, and it will contribute to the application of asexual reproduction to agriculture.
FOOD SECURITY: INNOVATIVE PLANT REPRODUCTIVE STRATEGY IN A CHANGING ENVIRONMENTAL CONDITION / C. Banfi ; scientific tutor: L. Colombo ; course coordinator: S. Ricagno. Dipartimento di Bioscienze, 2025 Feb 27. 37. ciclo, Anno Accademico 2023/2024.
PER UNA POETICA DELLE EMOZIONI. UNA RILETTURA DELL'ANTROPOLOGIA STÜRMERIANA ATTRAVERSO LE NEUROSCIENZE
"According to the neuroscientific theory of 4E Cognitive Science, emotions are defined as complex phenomena involving mind and body, guiding, and influencing all human actions. Therefore, in order to study them and investigate their diverse and numerous facets, it is necessary to adopt a transdisciplinary perspective combining different theoretical and methodological approaches, systematically and productively interweaving the humanities with the hard sciences. Situated within the framework of the most recent Neurohumanities, the main aim of this doctoral thesis is to bring literature into dialogue with neuroscience in order to help investigate the complexity of emotional phenomena. The roots of today’s neuroscientific theories on emotions, indeed, can be traced already in the anthropological idea of the ganzer Mensch and more specifically in the poetics that emerges from the analysis of autobiographical prose writing from the Sturm und Drang period, which can be delineated as a true ""Gefühlspoetik"". This poetics of emotions is traced by means of an original category of analysis, the ""Gefühlstextualität"", which represents the connecting point between the different aspects of the thesis and the key to the entire research. Indeed, through this category it is brought to light how emotions play a fundamental role both in literary expression, thus in the perspective of the ""Textimmanenz"", and in the experience of text reception, measurable in the Reader-response, which involves the possibility of a continuation of the research in an experimental direction. The theme of the emotions thus represents fertile ground for bringing together science and art, words and thought, body and mind, author and reader, past and present, while opening future research perspectives on the most intimate and unexplored depths of the human being, inhabited by fascinating creative and cognitive processes. In this sense, then, literature presents itself as a precious and powerful anthropological device."
PER UNA POETICA DELLE EMOZIONI. UNA RILETTURA DELL'ANTROPOLOGIA STÜRMERIANA ATTRAVERSO LE NEUROSCIENZE / I. Orlandazzi ; tutor: R. Maletta ; co-tutor: F. Caruana, G. Pulvirenti ; coordinatrice: L. Pinnavaia. Dipartimento di Lingue, Letterature, Culture e Mediazioni, 2024 May 27. 36. ciclo, Anno Accademico 2023.
SUBSTANTIAM REI MUTARE. FISICA ARISTOTELICA ED AZIONE MIRACOLOSA NELLA FONDAMENTAZIONE DUECENTESCA DELLA PLENITUDO POTESTATIS
"The goal of the thesis is to investigate the thirteenth-century origins of the concept of ""plenitudo potestatis"" (plenitude of power) by utilising the tools provided by political theology and adopting a legal-ontological perspective. Following this path, after an introduction that outlines some concepts employed in the research, the first topic examined is the medieval reception of Aristotle's physical texts, analysing the parallels between the teaching prohibitions imposed on these texts and those affecting Roman law. Building on this ground, the analysis explores the potential influence of these new conceptual frameworks on the development of the notion of ""miracle"" within scholastic thought. Additionally, it examines how this understanding of divine action aligns with the dichotomy between ordained (""ordinata"") and absolute power of God (""absoluta potentia Dei""), particularly as seen in the thoughts of Thomas Aquinas. With the philosophical and theological framework established, the thesis then delves into the evolution of the concept of ""plenitudo potestatis"", aiming to determine how it became associated with the idea of absolute divine power in the mid-thirteenth century. In the final section, the thesis focuses on the most significant claim tied to ""plenitudo potestatis"", namely, the Pontiff's power to ""change the substance of a thing"" (""substantiam rei mutare""). By studying thirteenth-century doctrine, the thesis ultimately reconstructs how these assertions of power were resolved through the interpretive faculties of power and the application of the ""canonical"" fiction of law (""fictio iuris"")."
SUBSTANTIAM REI MUTARE. FISICA ARISTOTELICA ED AZIONE MIRACOLOSA NELLA FONDAMENTAZIONE DUECENTESCA DELLA PLENITUDO POTESTATIS / M. Castelli ; tutor: R. Bianchi Riva, E. Fusar Poli ; coordinatore: F. Poggi. Dipartimento di Diritto Pubblico Italiano e Sovranazionale, 2023 May 31. 35. ciclo, Anno Accademico 2022.
EFFECT OF MICROTUBULE-TARGETING AGENTS ON STRUCTURAL SIGNALING AND CHEMICAL TRANSPORT IN CELLS
Microtubules, along with actin and intermediate filaments, are key components of the cellular cytoskeleton and play pivotal roles in several biological functions, including intracellular trafﬁcking, positioning of cellular components in interphase, mitotic spindle formation during cell division, maintaining of cell morphology and cell motility. Structurally, microtubules are formed by αβ-tubulin heterodimers that polymerize in a head-to-tail fashion to create protofilaments, which subsequently assemble into hollow cylindrical polar filaments. Microtubules’ behavior and function depend on their polymerization dynamics, with the first layer of regulation being the GTPase cycle of tubulin and its addition to the growing microtubule end. This process causes tubulin to transition from a curved conformation in solution to a straight conformation in the microtubule lattice. In cells, secondary layers of regulation (associated proteins, post-translational modifications, and tubulin isotypes levels) allow for a more precise modulation and differentiation of microtubule activities. Due to their involvement in key cellular activities, compounds that interfere with the microtubule cytoskeleton have been developed as cytotoxic agents, demonstrating remarkable efficacy against a wide spectrum of cancer types. Paclitaxel, a natural product extracted from yew tree bark, was the first microtubule-targeting agent (MTA) to be approved for clinical use in 1992, followed by two synthetic derivatives, docetaxel and cabazitaxel. These molecules bind to the taxane binding site in the microtubule, inducing its structural stabilization and preventing depolymerization events. Although the ability of taxanes to disrupt microtubule dynamics is closely linked to their chemotherapeutic effects, the exact cellular mechanism behind their anticancer activity remains under debate. At present, taxanes remain among the first-line treatments for several solid tumors, including breast cancer, ovarian carcinoma, pancreatic adenocarcinoma, non-small lung cancer, and prostate cancer. However, the clinical use of these molecules is limited by taxane-induced peripheral neurotoxicity, which severely affects up to 30% of patients and frequently leads to therapy discontinuation.To protect neurons from damage during taxane chemotherapy, it is essential to gain an improved mechanistic understanding of these drugs’ pathological effects on neurons. Due to their complex and elongated structures, neurons rely on intracellular active transport for cargo movement along the axon, making them especially susceptible to deficits in this process caused by taxane treatment. A recent hypothesis links axonal transport impairment to lattice expansion, a structural alteration induced by taxanes that appears to be unrelated to the stabilization effect. Locally expanded dimers may act as lattice point defects and cause premature detachment or stalling of the microtubule-associated motor proteins. Moreover, since current models of the microtubule structure describe the expanded lattice as a distinct structural feature localized near the tip, the spread of this structural signal along the whole lattice could impact the recognition patterns of motor proteins. Interestingly, unpublished results from our laboratory show that flutax-2, a C7-modified fluorescent taxane, does not induce microtubule lattice expansion. The present work, titled “Effects of microtubule-targeting agents on structural signaling and chemical transport in cells” focuses on two primary objectives. First, it aims to investigate the influence of C7-modifications of the taxane core on the cellular, biochemical, and structural properties of microtubules, in particular on lattice axial expansion. Second, it seeks to describe the relationship between the microtubule structural modifications induced by taxanes and the impairment of intracellular transport. These objectives wer
EFFECT OF MICROTUBULE-TARGETING AGENTS ON STRUCTURAL SIGNALING AND CHEMICAL TRANSPORT IN CELLS / F. Bonato ; tutor: J. F. Díaz Pereira ; tutor: M. Á. O. Blanco ; tutor: D. Passarella. Università degli Studi di Milano, 2023 Dec 19. 35. ciclo, Anno Accademico 2022.
DEEP LEARNING APPLICATIONS TO PARTICLE PHYSICS: FROM MONTE CARLO SIMULATION ACCELERATION TO PROTODUNE RECONSTRUCTION
"The thesis arises in the context of deep learning applications to particle physics. The dissertation follows two main parallel streams: the development of hardware-accelerated tools for event simulation in high-energy collider physics, and the optimization of deep learning models for reconstruction algorithms at neutrino detectors. These two topics are anticipated by a review of the literature concerning the recent advancements of artificial intelligence models in particle physics.Event generation is a central concept in high-energy physics phenomenology studies. The state-of-the-art software dedicated to Monte Carlo simulation is often written for general-purpose computing architectures, which allow great flexibility but are not compatible with specialized accelerating devices, such as Graphics Processing Units.The original tools presented in the thesis, PDFFlow and MadFlow, manage to combine these two aspects in Python and require no prior knowledge of specific programming languages for hardware accelerators. The former product, PDFFlow, is a Parton Distribution Functions interpolator, the latter, MadFlow, aims at building a complete tool suite to accelerate the whole event generation framework.The reconstruction pipeline at neutrino detectors is comprised of many different algorithms that work in synergy to extract a high-level representation of detector data. All the most important experiments in neutrino physics are developing software to automatically process and extract this information. This work describes the implementation of deep learning techniques to improve neutrino reconstruction efficiency at the ProtoDUNE-SP detector.Two original contributions are presented concerning raw data denoising and a hit-clustering procedure named ""slicing"". Both denoising and slicing involve the implementation and the training of novel neural network architectures, based on state-of-the-art models in machine learning, such as feed-forward, convolutional and graph neural networks. They represent a proof of concept that these models are indeed capable of providing an important impact on signal reconstruction at neutrino detectors."
DEEP LEARNING APPLICATIONS TO PARTICLE PHYSICS: FROM MONTE CARLO SIMULATION ACCELERATION TO PROTODUNE RECONSTRUCTION / M. Rossi ; referee: G. Panico; referee: D. Rousseau ; supervisor: S. Carrazza ; coordinatore: M. Paris. Dipartimento di Fisica Aldo Pontremoli, 2023 Feb 06. 35. ciclo, Anno Accademico 2022.
DISEASE RISK ASSESSMENT OF INVASIVE ALIEN SPECIES
Increased global trade and travel have led to a rise in the number of invasive alien species (IAS), i.e. species introduced by humans in geographic areas where are not naturally found, worldwide. Despite the recognized role of wildlife, as well as of wildlife translocations, in the emergence and re-emergence of infections of public health significance, IAS remain mainly studied for their environmental impacts, and their disease risk towards humans and animals is still largely neglected by health professionals. The main aim of this thesis is therefore to cover this gap by setting the ground for a new “invasion epidemiology” field, and this has been done through two main steps: the review and analysis of both the mechanisms underlying IAS disease risk and the information available in literature on IAS pathogens, and the development of a standardized qualitative disease risk assessment method, applicable to different geographic contexts, to assess the risk of mammal IAS to impact on human and animal health.First, I reviewed the existing biological and ecological literature on IAS to identify the main mechanisms by which animal IAS may affect disease risk in their area of release. IAS resulted to potentially affect disease risk both directly, by acting as hosts of infectious agents, thus possibly leading to the introduction of new pathogens, and/or the amplification of endemic ones, or indirectly, by altering the ecosystem equilibrium, through competitive and trophic interactions with native host species or the modification of local habitats. This literature review highlighted how IAS may have important health implications, which should be better acknowledged by people working in the human and animal health field, and how the mechanisms underlying the sanitary outcome of a biological invasion, and in particular indirect ones, are extremely complex, being the product of multiple factors. Acknowledging the important limitations of our current ability to predict possible health impacts driven by indirect mechanisms, I decided to address the issue of IAS disease risk by focusing specifically on IAS possible role as infectious agents’ host. As information on IAS pathogens is not systematized, preventing from knowing the amount and quality of available data to inform possible disease risk assessment procedures, I systematically reviewed the literature on the infectious agents of the main mammal species of European Union concern. Current knowledge on the pathogens harbored by mammal IAS was evaluated through different statistical approaches: the identification of the main factors associated with research intensity and the observed pathogen species richness, the estimation of the true helminth species richness, and a meta-analysis of prevalence of the pathogens of public and animal health significance. Results highlighted the existence of strong information gaps and biases in the way research on mammal IAS pathogens is carried out, the current underestimation of the amount of pathogens harbored by these species and high levels of uncertainty in the pooled prevalence of pathogens of public and animal health significance. However, the review confirmed that mammal IAS harbor pathogens of human and animal health significance, and therefore, the need to identify high-risk species. Considering that the existing knowledge gaps would have resulted in strong limitations in informing a risk assessment procedure, I developed a qualitative disease risk assessment methodology informed by expert opinion. This tool is specifically aimed at assessing IAS disease risk towards humans, domestic animal populations, and/or wildlife populations and allows to obtain a list of the pathogens of animal and human health significance that mammal IAS could transmit to a population of interest (directly or through the communities of local hosts), each with the related level of risk and uncertainty. Key feature
DISEASE RISK ASSESSMENT OF INVASIVE ALIEN SPECIES / E. Chinchio ; tutor: N. Ferrari ; co-tutor: M. Crotta ; coordinatore: C. La Vecchia. Dipartimento di Medicina Veterinaria, 2021 Mar 10. 33. ciclo, Anno Accademico 2020. [10.13130/chinchio-eleonora_phd2021-03-10].
ON THE CONTROL OF OPEN QUANTUM BATTERIES
Quantum control techniques provide a powerful framework to harness the potential of quantum technologies while addressing limitations imposed by noise and experimental constraints. These techniques enable critical tasks such as noise mitigation, dynamical shortcut engineering, and measurement optimization, with broad applications in quantum metrology, quantum state engineering, quantum computation, and quantum thermodynamics. In this thesis, we investigate how measurement-based quantum control protocols can enhance the thermodynamic performance of open quantum batteries (OQBs) across three complementary studies, focusing on noise resilience and operational efficiency in realistic, non-idealized scenarios.The first study characterizes the role of noise, particularly non-Markovian noise, in degrading the performance of OQBs. We demonstrate that environmental memory effects may play a non-trivial role in enhancing or degrading battery performance, particularly in charger-mediated interactions. The second work extends the concept of measurement-enhanced work extraction (daemonic ergotropy) to continuously monitored OQBs. We derive fundamental bounds on extractable work under both ideal and non-ideal conditions, establishing a theoretical framework for evaluating work extraction in continuously monitored quantum systems. In the third study we analyze the energy discharging from a quantum battery through a non-ideal quantum non-demolition (QND) energy measurement protocol, both in terms of work extracted and in terms of net energy balance, quantifying trade-offs between work gain and measurement costs. We show that, provided the extraction uses unitary operations and measurement efficiency exceeds a critical threshold, a net energy gain is possible.These works explore complementary aspects of OQB control, collectively advancing the understanding of quantum energy storage in noisy environments. The first characterizes context-dependent noise impacts, the second provides tools to benchmark measurement-based control, and the third identifies viability conditions for imperfect measurement-based extraction protocols. This work bridges theoretical quantum thermodynamics with experimentally relevant constraints, offering actionable insights for the design of noise-resilient quantum batteries. By advancing active control strategies tailored to non-Markovian environments and imperfect measurements, our results contribute to the development of robust quantum energy storage systems for near-term quantum technologies.
ON THE CONTROL OF OPEN QUANTUM BATTERIES / D. Morrone ; supervisore: M. G. Genoni ; coordinatore: A. Mennella. Dipartimento di Fisica Aldo Pontremoli, 2025 Mar 14. 37. ciclo, Anno Accademico 2023/2024.
EFFECTS OF LIPID MIXTURE AND A SELECTIVE PPARG MODULATOR ON THE DIFFERENTIATION CAPABILITIES OF HUMAN DERIVED MESENCHYMAL STEM CELLS(HADSCS) DERIVED FROM HEALTHY AN D BREAST CANCER PATIENTS
Metabolic syndrome is associated with many complications especially leadingto life threatening disorders such as obesity and cancer. To be able to identify solutionsand natural treatments, we need to investigate the underlying causes of this syndrome.Nutrition is one important factor to consider in the prevention and treatment of themetabolic syndrome. Nutrition effects almost all metabolism mechanisms in the humanbody. One provident effect of nutrition is adiposity. Over the recent years, an interestwas noted to studying adipogenesis in relation to obesity. Different factors affectadipogenesis including natural dietary compounds to help decrease adiposity, thereforethe risk of developing obesity and later on obesity related diseases such as breast cancer.To be able to study this correlation in-vitro, a wide choice of cell models can be used.Human adipose derived mesenchymal cells (hADSCs) are one of the top choices used tostudy adipogenesis overcoming the limitations that other cell models have in theirapplicability to humans regarding the prevailing difference in their metabolism andphysiology. In this study, the aim was to study adipogenesis using hADSCs in presenceof dietary compounds such as lipids and GMG-43AC, a natural selective peroxisomeproliferator-activated receptor g (PPAR g) modulator, that seems to have a positive effecton inhibiting adipogenesis in murine 3T3-L1 cells. We wanted to investigate further onits application on human cell models and try to understand its mechanism in inhibitingthis phenomenon. The protocols were set up using the THP-1 cell line, which we noticedupon using a Lipid mixture cocktail (Composition: Non-animal fatty acids; 2 μg/mlarachidonic; 10 μg/ml linoleic acid; 10 μg/ml linolenic acid: 10 μg/ml myristic acid; 10μg/ml oleic acid; 10 μg/ml palmitic acid; 10 μg/ml stearic acid; 0.22 mg/ml cholesterolfrom New Zealand sheep′s wool; 2.2 mg/ml Tween-80; 70 μg/ml tocopherol acetate), adecrease in pro-inflammatory cytokines IL-6 and IL-1b. We also noticed a doseIVdependent increase of FABP-4. Our findings regarding hADSCs, that PPARγ expressionand lipid accumulation was restored upon the presence of lipid mixture in breast cancerhADSCs that were derived from breast tissue. Secondly, GMG-43AC in bothconcentrations (0.5mM and 2mM) inhibited lipid accumulation and showed a significantdecrease in the expression of adipocyte-specific genes, such as PPARγ and FABP-4 evenafter the full differentiation of hADSCs that were derived from lipoaspirates. Thissuggests that dietary compounds are important factors in adipose differentiation and diethas a big influence in the progression and prevention in many metabolic diseases, such asobesity and cancer.
EFFECTS OF LIPID MIXTURE AND A SELECTIVE PPARG MODULATOR ON THE DIFFERENTIATION CAPABILITIES OF HUMAN DERIVED MESENCHYMAL STEM CELLS(HADSCS) DERIVED FROM HEALTHY AN D BREAST CANCER PATIENTS / G. Al Haj ; tutor: A. Barassi ; co-tutor: S. Carelli ; coordinator: L. Pinotti. Università degli Studi di Milano, 2020 Dec 09. 33. ciclo, Anno Accademico 2020. [10.13130/al-haj-ghina_phd2020-12-09].
L'ABUSO DI DIRITTO NEL SISTEMA EUROPEO COMUNE DI ASILO
This doctoral thesis aims to analyze the legal category of the prohibition of abuse of rights within the complex framework of European Asylum law and the relevant case law on the matter. The main objective of the research is to ascertain the scope of this concept within the Common European Asylum System (CEAS), to assess whether it can be established and legitimately justified. To do so, the research activity has been focused, first, on the notion of “abuse of rights” as elaborated by the European Court of Justice and as qualified by the relevant literature, with reference to other EU Law areas other than the CEAS. Through the analysis of this notion in different EU Law areas, I established a classification method that allowed me to categorize various abusive conducts in the wider framework of European Asylum law. Specifically, the classificatory method has been based upon i) the characteristics of the practices qualified as “abusive”; and ii) the legal qualification variously attributed to the prohibition of the abuse of rights. At the same time, I considered relevant for the research an accurate definition of the right to asylum, evaluated both from the perspective of international law, and in the light of the secondary law instruments adopted within the framework of the Common European Asylum System. This in-depth analysis has been necessary considering the complexities that characterize this right, both from a subjective point of view, with reference to the identification of the holder of such a legal position, and from an objective point of view, with respect to the definition of its content. Finally, I addressed the issue of the prohibition of abuse within the context of the Common European Asylum System by applying the classification method elaborated above. First, I identified the references to the notion of “abuse” within the EU Asylum legal framework and the jurisprudence of the Court of Justice of the EU. Then, I tried to draw potential conclusions on the qualification and the legal significance of this concept. An issue accompanied the entire analysis: since different notions of the right to asylum can be considered – which alternatively identify the State or the individual as the right holder –, it is necessary to identify who is actually responsible for the abuse: the asylum seeker or, rather, the State.
L'ABUSO DI DIRITTO NEL SISTEMA EUROPEO COMUNE DI ASILO / E. Colombo ; tutor: A. Lang ; coordinatore: F. Biondi. Dipartimento di Studi Internazionali, Giuridici e Storico - Politici, 2023 Jul 04. 35. ciclo, Anno Accademico 2022.
DESIGN AND DEVELOPMENT OF AN ADVANCED EXER-GAME BASED AUTONOMOUS HOME REHABILITATION PLATFORM
As life expectancy increases, we are witnessing continuous aging of the population and consequently an increase in age-related diseases. Also for this reason, the costs of health care and rehabilitation continue to rise, forcing hospital facilities to reduce the period of hospitalization, preventing patients from receiving proper treatment and potentially losing the motor skills acquired so far. To try to mitigate this problem, autonomous home rehabilitation has recently begun to spread, allowing patients to practice independently, reducing the workload and costs for hospitals and clinics. However, due to the absence of the therapist, several issues arise regarding the effectiveness of rehabilitation and patient safety.The aim of this research is to explore the critical aspects related to autonomous home rehabilitation and the recently proposed state-of-the-art platforms, and to develop a comprehensive system that allows patients to perform rehabilitation safely and effectively in full autonomy.In the project presented here, particular attention has been paid to the long-term motivation of the patient, with the implementation of exer-games, i.e., exercise through video-games, and other game design techniques, such as the dynamic adjustment of the difficulty. A platform dedicated to therapists is available, giving them all the tools they need to manage their patients, such as the possibility to schedule customized exercises and review the exercises performed. In addition, we have also explored how new mixed reality technologies could be used in rehabilitation and integrated into home rehabilitation platforms.We conclude with the preliminary results of two pilot studies in which 13 patients affected by multiple sclerosis and 24 healthy elderly participated. These tests have demonstrated the goodness and usability of the platform developed, but also the need for further trials with a larger number of patients to validate the proposed solutions and their long-term effectiveness.
DESIGN AND DEVELOPMENT OF AN ADVANCED EXER-GAME BASED AUTONOMOUS HOME REHABILITATION PLATFORM / M. Pezzera ; supervisor: N. A. Borghese ; coordinator: P. Boldi. Dipartimento di Informatica Giovanni Degli Antoni, 2021 Mar 22. 33. ciclo, Anno Accademico 2020. [10.13130/pezzera-manuel_phd2021-03-22].
MEDITERRANEAN DIET RESHAPES PERIPHERAL SECRETOME AND LIPIDOME PROFILES IN PATIENTS WITH METABOLIC SYNDROME
Weight loss in patients with metabolic syndrome has positive effects on cardiovascular diseases and type 2 diabetes risk, but its effects on peripheral secretome and lipidome profiles are still poorly understood.In order to determine the effects of diet-induced weight loss on metabolic parameters, lipidome and secretome profiles were evaluated.In this study, 18 adult males with metabolic syndrome and BMI between 25 and 35 Kg/m2 were enrolled, and then subjected to a balanced hypocaloric Mediterranean diet for 6 months.The aim of the dietetic approach was to induce in patients a weight loss of at least 5% of the initial body weight.After weight loss, we observed a significant improvement in BMI, insulin, fasting blood glucose, HOMA-I, triglyceridemia, LDL, and HDL levels.The analysis of circulating lipoproteins showed a significant change in their composition. In particular, a massive transfer of triacylglycerols from HDL to LDL was observed. This result was associated with a significant reduction in peripheral pro-inflammatory cytokines, such as IL-6, TNF-α, IL-8, and MIP-1β. We also observed an interesting positive correlation among cytokines levels and peripheral levels of CETP (cholesteryl ester transfer protein), an enzyme with a key role in lipid metabolism.The results achieved suggest that weight loss obtained through the hypocaloric Mediterranean diet is associated with an improvement in peripheral lipidome and secretome profiles. Furthermore, this dietetic approach stimulated changes in lipoproteins composition.These results are fundamental to understand weight loss benefits and the mechanisms that may play a role in improving cardiovascular risk.
MEDITERRANEAN DIET RESHAPES PERIPHERAL SECRETOME AND LIPIDOME PROFILES IN PATIENTS WITH METABOLIC SYNDROME / L. Loreggian ; tutor: G.V. Zuccotti ; coordinatore: L. Pinotti ; supervisore: P. Fiorina ; revisore: D. Corradi; revisore: A. Secchi. Università degli Studi di Milano, 2020 Dec 09. 33. ciclo, Anno Accademico 2020. [10.13130/loreggian-lara_phd2020-12-09].
QUALIFICATION MISMATCH AND OCCUPATIONAL CHANGE: THE ROLE OF DEMAND, SUPPLY AND INSTITUTIONAL FACTORS IN EXPLAINING CROSS-NATIONAL VARIATIONS
The occupational structure of advanced economies has undergone significant changes over the past decades. On the demand side, technological change intensified and has been considered the main driver behind these transformations, with competing predictions concerning its impact on society. On the supply side, a very substantial educational expansion took place worldwide, allowing to cope with technological progress, boosting productivity and growth. The interaction between the two creates reasons for optimism through the upgrading of occupational structure, and pessimism due to the “hollowing out” of susceptible to automation middle-skilled jobs and the growth in low-skilled jobs. Furthermore, the imperfect match between demand and supply resulted in an increasing phenomenon of qualification mismatch and overeducation, questioning whether having higher education results in having a better job. However, the magnitude of these processes appears to vary across countries, with more promising upgrading trends in some countries and polarization in others. This suggests that a more detailed assessment of the role of demand, supply factors, and institutional characteristics might be important for drawing conclusions and developing policy remedies to address the challenges posed by rapid technological advancements.In this dissertation, I analyse how the occupational structure has changed in 29 European countries over the past 15 years, investigating whether the technological change has been skill-biased or routine-biased and how these changes in occupational structure affected the risk of overeducation for individuals with different levels of education. I do so by analysing different sources of data and primarily the European Labour Force Survey (EU LFS). In the first part of the thesis, I provide a comparative assessment of changes in occupational structure based on different dimensions of job quality and task content of occupations. This assessment is followed by a multivariate regression analysis analyzing whether the cross-country dynamics of occupational change are explained by the different labor market institutions. In the second part of the thesis, I focus on the relationship between occupational structure and overeducation. I analyse how the incidence of overeducation varied across occupations and task-based occupational groups from a cross-country perspective. Finally, using a mixed-effects linear probability model, I explore how occupational change affects the risks of overeducation for different educational groups.The findings suggest that changes in occupational structure can be simultaneously described as skill- and routine-biased, because the latter successfully explains the decline in the middle that the former does not account for. Routinization does not appear to preclude upgrading, and polarization is determined by the position of routine-intense occupations in the country-specific wage structure, explaining cross-country variation of occupational patterns. Stronger upgrading is noticeable in the countries with higher investments in R&D, education, training, and adult training. Stricter employment protection legislation for regular contracts and collective dismissals mitigates the “hollowing out” in the middle, while the impact of the minimum wage on employment in low-skilled jobs appears to be limited.The evidence further suggests that upgrading of occupational structure and upskilling were accompanied by a rising incidence of overeducation, particularly in the period 2003-2010. Although it slightly declined in the period 2011-2018, a higher incidence of overeducation persisted among the tertiary-educated workers, especially in lower-skilled mildly-routine service occupations with a high degree of interactive tasks. Furthermore, while the risk of overeducation slightly declined for medium-skilled, it has not only persisted but increased for tertiary educated
QUALIFICATION MISMATCH AND OCCUPATIONAL CHANGE: THE ROLE OF DEMAND, SUPPLY AND INSTITUTIONAL FACTORS IN EXPLAINING CROSS-NATIONAL VARIATIONS / A. Damaronak ; supervisor: S. BERTOLINI ; co-supervisor: C. BORGNA ; director of doctoral program: G. BALLARINO. Dipartimento di Scienze Sociali e Politiche, 2022. 34. ciclo, Anno Accademico 2021.
A PROPAGATION OF SINGULARITIES THEOREM AND A WELL-POSEDNESS RESULT FOR THE KLEIN-GORDON EQUATION ON ASYMPTOTICALLY ANTI-DE SITTER SPACETIMES WITH GENERAL BOUNDARY CONDITIONS
In this thesis we study the Klein-Gordon equation on asymptotically anti-de Sitter (aAdS) spacetimes with boundary conditions implemented by pseudodifferential operators (PDOs) of order up to 2. Using techniques of microlocal analysis and b-calculus, we prove two propagation of singularities theorems, one for pseudodifferential operators of non-positive order and one for PDOs of order 0 < k <= 2, and we establish a well-posedness result in certain twisted Sobolev spaces. In particular, we discuss the existence and uniqueness of the advanced and retarded fundamental solutions for the Klein-Gordon equation with prescribed boundary conditions and we characterize their wavefront set. At last, as a concrete application, we build the fundamental solutions for a massless Klein-Gordon equation on a static aAdS spacetime with admissible boundary conditions using the formalism of boundary triples.
A PROPAGATION OF SINGULARITIES THEOREM AND A WELL-POSEDNESS RESULT FOR THE KLEIN-GORDON EQUATION ON ASYMPTOTICALLY ANTI-DE SITTER SPACETIMES WITH GENERAL BOUNDARY CONDITIONS / A. Marta ; supervisori: L. Pizzocchero, C. Dappiaggi ; coordinatore: V. Mastropietro. Dipartimento di Fisica Aldo Pontremoli, 2022 Jan 27. 34. ciclo, Anno Accademico 2021. [10.13130/marta-alessio_phd2022-01-27].
LE AFFINITÀ ELETTIVE. IL RAPPORTO TRA MAFIA E CAPITALISMO IN LOMBARDIA
The dissertation analyzes the causes of the growing «mafia demand» in Lombardy, a region unanimously considered the «locomotive of Italy», which can boast a long and important civic and entrepreneurial tradition, but which in recent years has been affected by the first convictions for mafia association of some of its entrepreneurs.Recovering Pierre Bourdieu's concept of «elective affinities», the research work reconstructs the stages of Mafia roots in the socio-economic, political and cultural fabric of the region, particularly in the city of Milan. From a diachronic perspective, it investigates the genesis and evolution of the relationship between the mafia phenomenon and the Lombard economic ruling class, through an in-depth study of significant events within the historical periodization adopted («mafia sowing», in 1950s-1960s - «the great transformation», in 1970s - «hybridization», in 1980s-1990s - «symbiosis», in 2000s).Starting from the assumptions that «the mafia is not a cancer that has proliferated by chance on healthy tissue» and that «the real strength of the mafia is outside the mafia», the research has identified those «potentially kindred» elements of the Lombard entrepreneurial culture and investigated the historically relevant causes that have allowed the various forms of the mafia phenomenon to become, in the span of 70 years, quasi-normal social actors of the Lombard socio-economic and political scenario.
LE AFFINITÀ ELETTIVE. IL RAPPORTO TRA MAFIA E CAPITALISMO IN LOMBARDIA / P. Farina ; tutor: F. Dalla Chiesa ; coordinatore: F. Basile ; revisori: I. Sales, G. Muti. Università degli Studi di Milano, 2021 Jul 13. 33. ciclo
IMPACT OF KETOGENIC DIET TREATMENT IN GLUT1 DEFICIENCY SYNDROME: DISCOVERY OF MOLECULAR MECHANISMS RESPONSIBLE FOR PHENOTYPE CHANGES IN BOTH PEDIATRIC AND ADULT PATIENTS
GLUT1 deficiency syndrome (GLUT1-DS) is a rare, effectively treatable, neurometabolic disease. It is caused by loss-of function mutations in the SLC2A1 gene, encoding GLUT1 protein, which supplies glucose to the brain. A genetic defect in this transporter leads to cerebral energy deficiency, resulting in a complex spectrum of neurological symptoms, including seizures and movement disorders. Although many clinical and molecular features have been deeply studied, there are still some not completely understood aspects, including (i) clinical variability and wide spectrum of SLC2A1 variations, (ii) phenotype age-related changes, and (iii) response to ketogenic diet (KD) treatment, which are investigated in this research study through a multi-omics approach.Firstly, we explored the clinical, biochemical and molecular data of 27 pediatric patients with GLUT1-DS, identifying (1) different clinical phenotypes (from early- to late-onset), (2) 25 disease-related SLC2A1 variants of which 10 never reported before, (3) a mutational hotspots in exon 4, suggesting a vulnerable region for glucose uptake associated to clinical manifestations, and (4) strong correlations between variant types, clinical and biochemical severity patterns.Next, we analyzed the transcriptional profiles and isoform switches in GLUT1-DS patients, before and after KD, focusing on three comparisons: (a) adults versus children, both untreated; (b) children in pre- versus post-KD; and (c) responders versus non-responders to KD in GLUT1-DS children. Overall, enrichment analysis revealed a reduction in inflammatory pathways in GLUT1-DS patients who experienced clinical improvements in seizure frequency and severity, whether in response to KD or during adulthood. These processes involve type I IFN signatures, which have an impact on mitochondrial function and signaling, suggesting a specific role in GLUT1-DS pathogenesis. Supporting these suggestions, the detected lncRNAs are also related to inflammatory response, as RMRP and LINC02555, or to ketone biosynthesis and oxidative phosphorylation processes, like PAX8-AS1, for which emerges its potential regulatory role in KD response. Another significant molecular pathway contributing to mitigate clinical signs is the alternative splicing of ion channel genes, such as P2X7 receptor which is, in turn, liked to inflammasome pathway. As well as this, the methylome profiles, performed by ONT, revealed altered methylation patterns in similar genes, like GRIN3A, suggesting also in this analysis an ion channel signaling deregulation.Lastly, these novel and suggestive findings underline the complex interplay that transcriptional variations, isoform switching and methylation patterns may have in the phenotype age-dependent changes and response to KD in patients with GLUT1-DS.
IMPACT OF KETOGENIC DIET TREATMENT IN GLUT1 DEFICIENCY SYNDROME: DISCOVERY OF MOLECULAR MECHANISMS RESPONSIBLE FOR PHENOTYPE CHANGES IN BOTH PEDIATRIC AND ADULT PATIENTS / A. Mauri ; tutor: G.V. Zuccotti, C. Cereda ; coordinatore: F. Cheli. Dipartimento di Scienze Biomediche e Cliniche, 2025 Mar 26. 37. ciclo, Anno Accademico 2024/2025.
CONDIZIONI PERSONALI DEL SOCIO E SITUAZIONI SOGGETTIVE SOCIALI NELLA S.P.A.
"This research moves in a regulatory scenario where the ""typological"" identity of capital companies has been profoundly transfigured, a particularly clear phenomenon in limited liability companies, as amply highlighted in the most recent doctrinal studies. In fact, more recently, there is a tendency to point out that the “type” s.r.l. has by now lost some identity connotations, which characterized it in past seasons, distinguishing itself by now, essentially, as a multifunctional matrix, tendentially neutral - or in any case hybrid - from the perspective of the possible - profoundly different - implementations, in terms of the ""typology of realities"", of the entrepreneurial program chosen by the associates.In the limited liability company, on the other hand, even if following the outcome of the reforms of 2003 and 2014, the principle of ""capitalist"" proportionality between investment risk and management ""power"" has experienced a substantial and general crumbling (or in any case a reduction in quantitative thresholds contained for some sections of the discipline, such as, for example, the proportionality between the voting power and the number of categories of shares held), legal doctrine certainly continues to record a greater sharpness and stability of the features that distinguish its unfailing characteristics, due to an unavoidable need for coherence with the typological identity of the joint stock company.In reality, as we will try to demonstrate along the course of the investigation, through the in-depth study of some segments of its discipline (in particular, with regard to the right to vote, the institutions of subordinated voting under the condition referred to in 2351, second paragraph, the graduated vote or by ceilings pursuant to Article 2351, third paragraph of the Civil Code and the “loyalty shares” pursuant to Article 127-quinquies of the TUF, for other subjective social situations, the institutions of restriction or subjective selection of the circulation or redemption of the shareholding) it seems possible to recognize an ever clearer opening of the legislator in the direction of a greater space of ""personalization"" of the shareholding for the statutory autonomy of the shareholders. In particular, in the latter direction, we will try to support and demonstrate that it is generally permitted for statutory autonomy to satisfy the aforementioned requirement by subordinating the entitlement or the intensity of some subjective social situations to predetermined conditions (also ""atypical"" ), relating to events concerning a personal circumstance of the shareholders, provided that the latter are identified “per relationem” in an indefinite manner, without therefore encroaching on the field of particular rights ""ad personam"". And this, according to a point of view which, even if at the root it follows in the wake of the more recent instances which tend towards an overcoming (or even better for a ""differentiated"" approach according to the segments of the discipline considered) of the monolithic opposition between contractual visions or institutional aspects of the joint-stock company, it seems more easily approachable to the first pole.Once the discipline sectors of the s.p.a. where it is possible to recognize the admissibility of the technique in question, a taxonomy is then proposed according to the ""typology"" of subjective conditions in question, and so to the principles of company law most directly involved; therefore, more generally, to the identifiable limits for the technique of differentiating the position of partners in speech, both from a static and dynamic perspective.Ultimately, therefore, following the outcome of the investigation, the effective typological physiognomy of the joint-stock company will be discussed, arriving at the conclusion that the phenomenon of differentiation of the mandatory characteristics of the type of capital companies, among"
CONDIZIONI PERSONALI DEL SOCIO E SITUAZIONI SOGGETTIVE SOCIALI NELLA S.P.A / G. Gitti ; tutor: M. Cian, M. Rescigno ; coordinatore: M. Rescigno. Dipartimento di Scienze Giuridiche Cesare Beccaria, 2023 May 05. 35. ciclo, Anno Accademico 2022.
DINAMICA DEI POTERI E FORMANTI ATIPICI DEL DIRITTO ECCLESIASTICO ITALIANO
The research aims to investigate how Law and Religion matters evolve in the continuing inertia of the legislature, with special attention to the links between the alterations in the order of sources and the changes affecting the functioning of the form of government.The work is organized into four chapters.The first chapter follows a recognitive approach, aimed at tracing, from a diachronic perspective, the evolution of the sources (formants) of Italian Law and Religion, in light of the contextual changes in political-institutional balances.The second chapter aims to reconstructing the current and complex physiognomy of the Italian Law and Religion system.The third chapter adopts a broader perspective, drawing on the reflections that have engaged the whole public law for decades, in an attempt to provide an answer to the question of whether the changes that are affecting Law and Religion are (or are not) part of a broader general trend affecting the legal system in its entirety.The fourth and final chapter returns to Law and Religion matters by noting first of all how, in light of the changed systemic regime, the idea of a general law on religious freedom appears essentially utopian.
DINAMICA DEI POTERI E FORMANTI ATIPICI DEL DIRITTO ECCLESIASTICO ITALIANO / G. Pavesi ; tutor: M. Toscano ; coordinatrice: F. Poggi. Dipartimento di Scienze Giuridiche Cesare Beccaria, 2025. 37. ciclo, Anno Accademico 2023/2024.
MEASUREMENT OF THE MAGNETIC AND ELECTRIC DIPOLE MOMENTS OF THE $\LAMBDA$ BARYON AT LHCB
One of the most profound and unresolved questions in modern physics is why the Universe is composed predominantly of matter, with negligible antimatter. This imbalance suggests the existence of processes that violate the fundamental symmetry between matter and antimatter, the so-called CP symmetry. However, the amount of CP violation predicted by the Standard Model of particle physics (SM), via the Cabibbo-Kobayashi-Maskawa (CKM) quark-mixing mechanism, cannot explain the large observed asymmetry, thus new sources of CP violation, beyond the SM, are expected. A possible way to probe such sources is to search for electric dipole moments (EDMs) of fundamental particles. This thesis explores that possibility through the study of the \Lz baryon: an EDM observation with the current sensitivities would constitute evidence of CP violation beyond the SM. Moreover, the magnetic dipole moment (MDM) of the \Lz baryon and the $\bar{\Lz}$ antibaryon will be measured simultaneously, allowing an unprecedented test of the CPT symmetry in this particle-antiparticle system. Notably, the MDM of the $\bar{\Lz}$ antibaryon has never been measured to date. All these measurements of the \Lz baryon electric and magnetic dipole moments are performed for the first time at a collider experiment.The analysis is conducted using data collected by the LHCb experiment during the \mbox{Run 1} and \mbox{Run 2} of the LHC, corresponding to an integrated luminosity of about 9 fb$^{-1}$. The LHCb detector was designed to investigate CP-violating phenomena of beauty and charm hadrons. This thesis describes how its capabilities have been extended to enable investigations of dipole moments of strange baryons. The measurement relies on the spin precession of longitudinally polarized \Lz baryons in the magnetic field of the LHCb tracking system. The polarization is inferred via an angular analysis of the \Lz decay products. A novel aspect of this work is the use of particles that decay downstream of the LHCb magnet, reconstructed solely with hits in the final tracking stations. These so-called T tracks present significant challenges in the reconstruction due to their limited momentum and vertex resolutions. Their exploitation in physics analyses is being pioneered in the context of this study.To overcome the performance limitations, a dedicated study to improve the reconstruction of long-lived \Lz and \KS decays using T tracks has been conducted in the first part of the thesis, developing custom techniques with significant impact on the reconstruction efficiency and resolutions. In the second part, the measurement of the electric and magnetic dipole moments is performed using \LbToJpsiLz decays. The analysis is validated using \BdToJpsiKS decays as control sample, in which the long-lived $K_S^0$ meson, being a pseudoscalar particle, does not exhibit any polarization nor dipole moments. The measurement of dipole moments is performed on the control sample as a null test to validate the method before applying it to the signal decays.The LHCb detector has undergone a major upgrade for the data-taking during Run 3 of the LHC. In this context, the new detector is briefly presented, with a particular focus on the Upstream Tracker. The author of this thesis contributed to this subsystem during its assembly, installation, and commissioning phases.
MEASUREMENT OF THE MAGNETIC AND ELECTRIC DIPOLE MOMENTS OF THE $\LAMBDA$ BARYON AT LHCB / G. Tonani ; supervisor: N. Neri, F. M. Vidal ; coordinatore: A. Mennella. Dipartimento di Fisica Aldo Pontremoli, 2025 Sep 30. 37. ciclo, Anno Accademico 2024/2025.
ASSOCIAZIONE TRA SCORE GENETICO GRS E RISCHIO DI EPATOCARCINOMA IN PAZIENTI CON CIRROSI HCV TRATTATI CON FARMACI ANTIVIRALI DIRETTI
Background and aim: Several single nucleotide polymorphisms (SNPs) have been associated with hepatocellular carcinoma (HCC) in hepatitis C virus (HCV) cirrhotics, however their role in patients cured by direct-acting antivirals (DAA) is still undefined. We assessed the association between a genetic risk score (GRS) based on the combination of 4 SNPs (PNPLA3 rs738409, MBOAT7 rs641738, TM6SF2 rs5842926 and GCKR rs1260326) and HCC in a cohort of DAA-treated patients. Methods: Consecutive HCV cirrhotics receiving DAA between December 2014-2016 in a single Center were genotyped for the 4 SNPs. Cirrhosis was defined histologically or non-invasively (Liver stiffness measurement [LSM] ≥12 kPa). HCC was diagnosed and staged according to international recommendations. Results: 509 patients were analyzed: median age 64 (28-87) years, 58% males, LSM 19.4 (12.0-75.0) kPa, 87% Child-Pugh score A (CPT) A, 11% with previous HCC history. Genotypes distribution was as follows: PNPLA3 CC (46%), CG (41%) GG (13%); MBOAT7 CC (29%), CT (49%), TT (22%); TM6SF2 CC (91%), CT (8%), TT (1%); GCKR CC (26%), CT (48%), TT (26%). Median GRS score in the overall population was 0.3 (0-1.1). Patients’ demography and clinical features were similar across SNPs genotypes. Following antiviral treatment, 491 (96%) patients achieved viral eradication. During a median follow-up of 43 (3-57) months from DAA start, HCC developed in 36/452 (8%) patients without a history of HCC (de novo HCC), 4-year estimated cumulative probability of HCC being 9% (95% CI 7-12%). Male sex (Hazard Ratio [HR] 2.54; 95% CI 1.15-5.63; p=0.02), diabetes (HR 2.39; 95% CI 1.20-4.74; p=0.01), albumin (HR 0.35; 95% CI 0.19-0.64; p=0.001) and GRS score >0.6 (HR 2.30; 95% CI 1.03-5.11; p=0.04) were independently associated with de novo HCC. Indeed, 4-year cumulative rates of de novo HCC resulted 6% (95% CI 1-10%) vs. 12% (95% CI 8-16%) in males vs. females (p=0.01); 17% (95% CI 9-24%) vs. 7% (95% CI 5-10%) in diabetic vs. non-diabetic (p=0.001); 21% (95% 10-34%) vs. 7% (95% CI 4-11%) in patients with albumin ≤ or >3.5 g/dl (p<0.001) and 16% (95% CI 8-28%) vs. 7% (95% CI 5-11%) in patients with a GRS score > or ≤0.6 (p=0.01), respectively. Main tumor features did not significantly differ according to the 4 SNPs genotypes. By combining independent risk factors for HCC, 4-year cumulative incidence resulted 20% (95% CI 12-28%) vs. 5% (95% CI 3-7%) in patients with or without two different risk factors, respectively (p<0.0001). HCC developed in 28/57 (49%) patients with a previous HCC history, corresponding to a 4-year cumulative HCC recurrence of 49% (95% CI 27-63%). At multivariate analysis, only diabetes (HR 2.77, 95% CI 1.15-6.67; p=0.02) was independently associated with recurrent HCC, while GRS score was not significant. Indeed, 4-year cumulative incidence of HCC resulted 88% (95% CI 64-100%) vs. 40% (95% CI 25-55%) in diabetic vs. non-diabetic patients, respectively (p=0.002).Conclusions: In a large, single-center cohort of consecutive HCV patients with cirrhosis who received DAA treatment, a genetic risk score was independently associated with de novo HCC, together with clinical predictors (male sex, diabetes, albumin values). Combination of clinical and genetic predictors could allow a better HCC risk stratification in HCV cirrhotic patients after viral cure, in order to develop personalized HCC surveillance programs.
ASSOCIAZIONE TRA SCORE GENETICO GRS E RISCHIO DI EPATOCARCINOMA IN PAZIENTI CON CIRROSI HCV TRATTATI CON FARMACI ANTIVIRALI DIRETTI / E. Degasperi ; tutor: F. Caprioli ; coordinatore del corso: E. Berti, P. Lampertico ; supervisori: M. Mondelli, P. Invernizzi. Dipartimento di Fisiopatologia Medico-Chirurgica e dei Trapianti, 2020 Mar 18. 32. ciclo, Anno Accademico 2019. [10.13130/degasperi-elisabetta_phd2020-03-18].
A MECHANISTIC APPROACH TO SYNAPTIC PLASTICITY AND COGNITION:SYNAPTIC STABILIZATION OF NMDARS THROUGH THE NOVEL INTERACTOR RABPHILIN-3A
The glutamatergic synapse is involved in the modulation of higher brain functions, such as learning and memory processes. The activation of specific subtypes of N-methyl-D-aspartate-type glutamate receptors (NMDARs) located at synapses accounts for different electrophysiological properties of neurotransmission and consequent synaptic plasticity mechanisms. NMDAR subunit composition is relevant for physiological neuronal functions and is often altered in many neurological disorders. In particular, augmented levels of synaptic NMDARs containing the GluN2A subunit can be observed after potentiation of neurotransmission during Long-Term Potentiation (LTP). Several studies revealed that LTP at hippocampal synapses underlies encoding and consolidation of memory.Rabphilin-3A (Rph3A) was recently characterized as a specific GluN2A intracellular binding partner promoting stabilization of GluN2A containing NMDARs at postsynapses through a trimeric complex with the main postsynaptic scaffolding protein PSD-95. Silencing of Rph3A or disruption of Rph3A/GluN2A/PSD-95 interactions leads to reduction in GluN2A-containing NMDARs due to receptor endocytosis. The modulation of this complex in the striatum of parkinsonian animals showing a dyskinetic behavior was able to restore normal locomotive behavior, indicating Rph3A could represent a new pharmacological target in brain diseases. However, the role of Rph3A in hippocampal functions such as synaptic plasticity and learning and memory has not yet been elucidated. In the present PhD project we aimed to investigate the role of Rph3A/GluN2A/PSD-95 complex in these events applying different stimulation protocols both in vitro and in vivo. In addition, we investigated the possible interplay between these synaptic complexes and the synapse-to-nucleus messenger Ring Finger Protein 10 (RNF10).We discovered that Rph3A is present in almost 50% of hippocampal dendritic spines at resting conditions. However, after potentiation of neurotransmission the number of Rph3A positive spines increases, paralleled by augmented formation of Rph3A/GluN2A/PSD-95 trimeric complex. Interference with Rph3A/GluN2A interaction through different experimental approaches leads to failure in LTP induction both at molecular and morphological levels, impairing also hippocampal dependent spatial learning. Dendritic spines displaying Rph3A show higher maturation degree in morphological parameters and recruit more newly-synthetized proteins compared to Rph3A negative ones, suggesting that Rph3A positive spines represent more stable and mature neuronal connections. Furthermore, the molecular motor transporter Myosin-VA was previously described as a binding partner of Rph3A and involved in GluA1-containing AMPA receptors delivery to synapses during synaptic potentiation. Interestingly, a trimeric complex composed by Rph3A/MyoVA/GluA1 is detected in hippocampus and their interaction is increased after cLTP application. Finally, our results show that impairing the synapse-to-nucleus transport of RNF10 during chemical-LTP isdetrimental for Rph3A synaptic localization, indicating the integration of synapse-to-nucleus signals during synaptic plasticity probably impacts on Rph3A postsynaptic functions.In conclusion, our results suggest that Rph3A postsynaptic role is not only given by the stabilization of GluN2A containing NMDARs accounting for adequate LTP induction and hippocampal learning process, but also for modulation of synaptic responsiveness through the delivery of GluA1-containing AMPA receptors during synaptic potentiation.
A MECHANISTIC APPROACH TO SYNAPTIC PLASTICITY AND COGNITION:SYNAPTIC STABILIZATION OF NMDARS THROUGH THE NOVEL INTERACTOR RABPHILIN-3A / L. Franchini ; main supervisor: F. Gardoni (Università degli Studi di Milano) ; external co-supervisor: C. Bellone ( Università di Ginevra) ; phd coordinator: A. L. Catapano. Dipartimento di Scienze Farmacologiche e Biomolecolari, 2021 Jan 13. 33. ciclo, Anno Accademico 2020. [10.13130/franchini-luca_phd2021-01-13].
EFFECT OF CIGARETTE SMOKING ON CARDIORESPIRATORY AND METABOLIC RESPONSE TO DIFFERENT PROTOCOLS IN YOUNG ACTIVE MALES
Cigarette smoking is one of the most impacting risk factors for cardiopulmonary morbidity and a major cause of mortality. It leads to systemic modifications that alter the cardiorespiratory and metabolic response at rest and during exercise. Indeed, stimulates the sympathetic nervous system, elevating the heart rate (HR) and cardiac work. Likewise, tar impairs O2 diffusion across the alveolar-capillary barrier, and carbon monoxide reduces O2 transport and extraction. The analysis of the cardiorespiratory and metabolic transitions at different work rates can provide insights in the O2 transport and delivery mechanisms. Hence, this dissertation aimed to provide a comprehensive examination of the effect of cigarette smoking on the cardiorespiratory and metabolic responses to exercise in young, physically active males. The smokers, compared to a no-smoker group, were evaluated in different exercise protocols: i) moderate intensity square-wave work rates to assess the cardiorespiratory and metabolic kinetics; ii) incremental stepwise protocols to determine the maximum oxygen uptake (V ̇_(O_2 max)), a benchmark for cardiorespiratory fitness, recovery kinetics, and exercise capacity; iii) moderate and iv) heavy intensity sinusoidal work rates. Cigarette smoking has detrimental effects on cardiorespiratory and metabolic response to different exercise protocols in young, physically active males. Despite young age, high fitness level and similar pulmonary function, the smokers were characterized by slower cardiorespiratory and metabolic kinetics during moderate exercise and during the recovery of an incremental test. Interestingly, there were no differences between the smokers and controls during both sinusoidal exercises, except for a shorter time to exhaustion that may suggest peripheral dysfunction.
EFFECT OF CIGARETTE SMOKING ON CARDIORESPIRATORY AND METABOLIC RESPONSE TO DIFFERENT PROTOCOLS IN YOUNG ACTIVE MALES / M. Borrelli ; advisor: F. Esposito ; phd coordinator: C. Sforza. Dipartimento di Scienze Biomediche per la Salute, 2023 Apr 18. 35. ciclo, Anno Accademico 2022.
LA FUNZIONE CONOSCITIVA DEL PARLAMENTO. NORME, PRASSI, PROCEDIMENTI
The aim of this research is to investigate whether the acquisition of knowledge, information and data can constitute an autonomous parliamentary function, not necessarily linked to other functions. In recent years, Parliaments have increased and developed particularly broad and penetrating fact-finding powers, hence the hypothesis that legislatures have a new and cross-cutting function. The greater complexity of the issues with which Parliaments are invested has given centrality to fact-finding tools, as the only ones available to all Members of Parliament, both from the majority and the opposition. The use of these tools often constitutes a moment of conciliation between the technical expertise and political decision. And today, in the age of information and big data, this becomes even more relevant, after the health and economic crisis experienced on a global scale. The purpose of the research is not merely to reach a comprehensive definition. On the contrary, after having formally defined “knowledge” as a parliamentary function, the aim is to examine the concrete practice of this function, examining the provisions of the parliamentary rules of procedures and praxis (codified or not). The review of the tools used in parliamentary activity is useful to understand if the proposed definition finds correspondence in the parliamentary rules and procedures, and how it systematically crosscuts other parliamentary functions, in particular the legislative and oversight ones, in accordance with the principle of multifunctionality of parliamentary procedures.The acquisition of “knowledge” requires the existence of a requesting actor (the Parliament), and of another actor who provides, in whole or in part, the requested information (the Government). In systems such as the Italian one, it is evidently an interlocutory moment proper to the confidence relationship between the Parliament and the Executive. For this reason, fact-finding parliamentary powers will be examined also with regard to the Government, in order to state if and how the exercise of the aforementioned function invests the prerogatives and functions of the Executive, also on an organizational basis.The concrete performance of the fact-finding activities in financial and budgetary procedures will then be examined. After all, the system of “checks and balances”, typical of parliamentary forms of government, takes shape in the assuming of financial decisions. When taking financial decisions, Parliament is required to respect certain constraints, and this implies the acquisition of data and information, so as to comply with these constraints. Thus, the preliminary activity carried out by the parliamentary committees is both legislative and financial, since the financial effects of legislative and non-legislative acts are examined.Methodologically, the research is conducted along two lines: the first is the theoretical one, aimed at formalizing “knowledge” as a parliamentary function; the second is the practical one, aimed at verifying the existence of this function on an empirical level. The first research segment engages with the traditional classifications of parliamentary functions, whereas the second research segment starts with parliamentary rules of procedures. The field of investigation is constituted by the Italian Parliament and the Italian legal system in general, considering both the Chamber of Deputies and the Senate of the Republic. The research also attempts to offer an updated view of the fact-finding tools, excluding from the analysis the scrutiny tools (questions, interpellations) and the parliamentary inquiries.
LA FUNZIONE CONOSCITIVA DEL PARLAMENTO. NORME, PRASSI, PROCEDIMENTI / F. Luciano ; tutor: F. Biondi ; coordinatore del corso: F. Biondi. Dipartimento di Diritto Pubblico Italiano e Sovranazionale, 2023 Jul 05. 35. ciclo, Anno Accademico 2022.
APPLICATION OF OMICS APPROACHES IN DRUG DISCOVERY AND BIOMARKER IDENTIFICATION
Recent advances in omics sciences have represented a revolution in many fields of biological research. Among these, lipidomics and metabolomics are gaining increasing interest due to their close connection with the organism’s phenotype. As the main analytical goal of omics approaches is to identify and quantify as many compounds as possible within a studied system, powerful analytical techniques and bioinformatics tools are required. Mass spectrometry (MS) coupled with liquid chromatography (LC) is one of the most powerful analytical platforms for the analysis of the lipidome in complex biological matrices and offers promising new insights in this field. Lipids are a large class of biomolecules involved in many biological processes with signalling, biophysical, and metabolic functions. The lipidome is complex, consisting of many species that share the same elemental composition but have different structural and physicochemical properties. Its comprehensive analysis is, therefore, technically challenging, and advanced LC-MS-based lipidomics workflows aim to better address this complexity. Lipidomics approaches can potentially be applied in all therapeutic areas, including cardiovascular, metabolic and inflammatory diseases. This is because an increasing number of human diseases are associated with significant lipidome remodelling, and changes in lipid profile have been identified as a major risk factor for many of them. With this thesis, we aimed to unravel the complexity of the lipidome and to assemble this knowledge in the description of biological systems to highlight lipid changes in pathology or response to drug treatment. To this end, we have optimized MS-based untargeted analytical workflows for the comprehensive profiling of lipids in different biological matrices and under various experimental conditions (in vitro and in vivo models) using advanced high-resolution LC-MS approaches.In Chapter 2, we integrated omics approaches into phenotypic screening drug discovery to support the identification of novel bioactive compounds. These studies aimed to apply untargeted multi-omics approaches to infer the mechanism of action of phytocomplex with potential lipid-lowering properties. Once better understood, lipid metabolism can be targeted pharmacologically. Phytochemicals from natural extracts represent an important source of bioactive compounds potentially useful in treating metabolic diseases characterized by major lipidome remodelling. However, the heterogeneity of natural matrices emphasizes the need for advanced analytical workflows to explore their beneficial effects, elucidating their mechanism of action and potential molecular targets. Using untargeted MS-based measurement of lipids and proteins allows a comprehensive investigation of the molecular mechanism of Scutellaria baicalensis (Study I) across different cellular states to help define the molecular changes responsible for its beneficial effect against liver disorders. In Study II, we established an analytical workflow to identify bioactive compounds in a polyphenolic fraction of bergamot leaves (Citrus bergamia). The experimental approach, which included a prefractionation step followed by LC-MS/MS analysis and phenotypic screening, revealed the great potential of leaves, usually considered as only waste products, as a source of polyphenols with positive effects on liver lipid metabolism. In Chapters 3 and 4, we applied UHPLC-MS analysis to study lipidome signatures in in vivo models focusing on specific key objects. Our primary goal was to determine the applicability of untargeted LC-MS/MS methods for the comprehensive analysis of lipid profiles in complex biological samples and to ensure that the analytical method was effective in capturing lipidome diversity and possible alterations with respect to environmental damage or pathological features. This helped us to derive some knowledge about how such stimuli
APPLICATION OF OMICS APPROACHES IN DRUG DISCOVERY AND BIOMARKER IDENTIFICATION / B. Zoanni ; tutor: M. Carini ; coordinatore: G. Vistoli. - Dipartimento di Scienze Farmaceutiche, via Mangiagalli 25, Milano. Dipartimento di Scienze Farmaceutiche, 2025 Jan 29. 37. ciclo, Anno Accademico 2023/2024.
NUTRITIONAL AND FUNCTIONAL CHARACTERISATION OF ALTERNATIVE FEED AND FOOD INGREDIENTS: EMPHASIS ON HEMP-BASED PRODUCTS, CO-PRODUCTS, MICROALGAE AND CULTURED MEAT
The growth of the world population has led to an increase in demand for food, particularly meat and dairy products, further intensifying livestock production, resulting in an enhanced use of grain for animal feed and competition with crops for direct human consumption. In parallel, over time, the livestock sector has exerted considerable pressure on resources, with large areas of arable land dedicated to food cultures and grazing, as well as high water use. This highlights the urgent need for action with multidisciplinary approaches. For this reason, the aims of this thesis were to evaluate the nutritional and functional potential of alternative plant-based products, including hemp, cardoon, and camelina, by analysing their antioxidant properties, bioactive peptide profiles, and their impact on the quality of animal-derived products. It also aimed to optimize the culture medium of microalgae, in particular for Euglena gracilis to enhance its functional profile while investigating the risks of nanoplastics and microplastics contamination during cultivation under heterotrophic conditions. Additionally, the research sought to address key challenges in cultured meat production by developing alternatives to fetal bovine serum, scalable biofabrication methods, and the integration of waste-derived components to support cell proliferation and differentiation. These objectives collectively aimed to advance sustainable, ethical, and innovative practices in the agri-food sector.Hemp based-products and their co-products have demonstrated high nutritional and functional profiles, as observed following in vitro digestion and solvent extraction methods. In support of this, an in vivo trial, conducted on laying hens, showed how the inclusion of a hemp co-product of up to 9% (leaves, non-standard hemp seeds, hulls and stems) improved performance while increasing the functional and nutritional profile of the eggs. However, it is important to emphasise that their applicability in the livestock sector requires a multidisciplinary approach, evaluating not only the positive effects on animal health but also taking into account limiting factors such as availability, high cost and variability.Among the various possible alternatives, microbial proteins, in particular microalgae, are strong candidates. In particular, as demonstrated in this thesis, the modulation of the culture medium of E. gracilis, cultivated under heterotrophic conditions, led to improvements in the nutritional and functional profile. This result represents a milestone in the formulation of a product that can be specifically designed for the needs of farm animals. However, the widespread use of plastic bioreactors for microalgae cultivation poses potential health challenges for the user. Indeed, plastics can release micro- and nanoplastics into the culture medium, which can accumulate in the biomass and enter the food chain, potentially impacting animal and human health.Finally, a possible alternative of recent interest is the production of cellular foods. In this field, cultured meat is certainly the most discussed. Although scientific research has tried to find answers to the main questions concerning the production process and its sustainability, many open issues still remain. Among these, alternatives to foetal bovine serum that can guarantee the growth, proliferation and differentiation of muscle cells through an ethical and sustainable process deserve further and necessary investigation. As demonstrated in this thesis, possible alternatives involve co-products of food (e.g. whey proteins), which can support the viability, proliferation and differentiation of C2C12 murine muscle cells.Although further studies are needed, all these alternatives represent fundamental steps towards the creation of resilient and environmentally friendly food systems. Despite promising results, further research is needed to standardise these approaches and address unresolved issues, ensuring
NUTRITIONAL AND FUNCTIONAL CHARACTERISATION OF ALTERNATIVE FEED AND FOOD INGREDIENTS: EMPHASIS ON HEMP-BASED PRODUCTS, CO-PRODUCTS, MICROALGAE AND CULTURED MEAT / D. Lanzoni ; supervisor: C. Giromini ; co-supervisor: E. Pěchoučková ; coordinator: F. Cheli. - Lodi. Università degli Studi di Milano, Dipartimento di Medicina Veterinaria e Scienze Animali. Dipartimento di Medicina Veterinaria e Scienze Animali, 2025 Mar 21. 37. ciclo, Anno Accademico 2024/2025.
PROS AND CONS OF NANOMATERIALS: STUDY OF HALLOYSITE-BASED NANOCOMPOSITES FOR BIOMEDICAL OR CATALYTIC PURPOSES AND EVALUATION OF ENVIRONMENTAL AQUATIC TOXICITY OF CEO2 NANOPARTICLES
My Ph.D. dealt mainly with the study of halloysite nanotubes (HNT), and their interaction with molecules and nanoparticles as potential for potential applications either in biomedicine or in catalysis. A minor part of my Ph.D work was dedicated to the preparation and characterization of Ceria nanoparticles stabilized with polymers of different charge, to investigate, in collaboration with the group of prof Della Torre and Binelli, their effect on various animal and bacterial models relative to their possible toxic effects especially in the aquatic environment.I started the first part of my thesis by introducing the clay material HNT. HNT is a unique natural nanomaterial composed of double-layered aluminosilicate with a hollow tubular structure in the micro range. A preliminary literature survey revealed that HNT, due to their physical and chemical features, could be suitable for many application fields like medicine and catalysis (Chapter 1). We started our aim by synthesizing some HNT adducts possibly suitable for hyperthermia applications by selectively load superparamagnetic iron oxides (SPIONs) into the inner lumen of HNT. The magnetic properties of the loaded SPIONs did not change after their trapping inside the HNT lumen. The SPION-in-HNT nanocomposite was synthesized through the pre-modification of the HNT inner lumen such that it becomes suitable to be suitable to load the apolar SPION as synthesized by thermal decomposition method without a further step of ligand exchange (Chapter 2). To extend the loading to another kind of NP for hyperthermia applications, we tried to load gold nanoparticles. The gold NPs can be synthesized in various shapes; we synthesized spherical and star shapes of gold for our purpose. Gold can be loaded inside the inner lumen in the studied shapes, but the low concentration of gold suspension hampered a massive loading into the HNT lumen. (Chapter 3). The second part of the work with Halloysites was devoted to the preparation and characterization of HNT adducts with luminescent molecules able to act as photosensitizers for photodynamic therapy (PDT). For this purpose, we were interested in loading perfluorinated porphyrin, in their non-coordinated- and Zn-coordinated form, inside the lumen of HNT. The release of the photosensitizer by the inner lumen was slowed down compared to the release of the perfluorinated drugs adsorbed in the outer surface of HNT. (Chapter 4). To use HNT as a dual vector for drug delivery, we synthesized HNT-Ru photosensitizer. The photosensitizer was covalently bonded to the silica part of the outer surface of HNT, leaving the inner lumen free for potentially an extra loading with another drug. The photophysical properties of synthesized nanocomposite were then tested (Chapter 5). Finally, concerning HNT, we aimed to synthesize a new synthetic Au-Pt nanoparticle supported over HNT with different Au-to-Pt molar ratios via a sol immobilization method for catalytic purposes. The synthesized catalyst showed that the activity toward hydrogenation of C=O of cinnamaldehyde increased with increasing the molar ratio of Au/Pt (Chapter 6). Unfortunately, the product potentially useful for biomedical purposes were not tested at least at cellular level, and this lack to the work was essentially due to the COVID-19 pandemic situation that slowed down a lot of collaboration especially with biologist collaborators.The last part of my thesis was devoted to synthesizing nanoceria and covering it with natural polymer available in the aquatic system as alginate and chitosan. Ceria NPs surrounded by both Alginate and Chitosan showed no acute toxicity effects at the average environmental concentration level of the two tested natural macromolecules (Chapter 7).
PROS AND CONS OF NANOMATERIALS: STUDY OF HALLOYSITE-BASED NANOCOMPOSITES FOR BIOMEDICAL OR CATALYTIC PURPOSES AND EVALUATION OF ENVIRONMENTAL AQUATIC TOXICITY OF CEO2 NANOPARTICLES / H.a.i. Hamza ; supervisor: D. MAGGIONI ; coordinatore: D. ROBERTO. Dipartimento di Chimica, 2021 May 11. 33. ciclo, Anno Accademico 2020. [10.13130/hamza-hady-ahmed-ibrahim_phd2021-05-11].
«ICH BIN EIN REGISSEUR DES WORTS UND NICHT DER DEKORATION»:L¿ESTETICA TEATRALE DI LEOPOLD JESSNER: QUASI UN ÓRGANON
This dissertation aims at reconstruing Leopold Jessner’s theatre aesthetics through theanalysis of his theatrical essays. Jessner was one of the most influential German theatredirectors during the Weimar Republic. His figure came along in the cultural landscape in thesame years the director’s theatre emerged in both Europe and Germany. Through theanalysis, this dissertation also aims at rethinking Jessner’s theatre aesthetics in the artistic andcultural landscape of the Weimar Republic.The second chapter of the dissertation presents Jessner as a theatre director and focusseson the obstacles and labels that prevented a scientific evaluation of his work.The third chapter proposes an analysis of the author’s corpus. In order to shed light on thekey aspects of Jessner’s aesthetics, the investigation is organised in four macro-sections: DerTheaterleiter, Zeittheater und Theaterpolitik, Regie und Schauspielkunst and Betrachtungen undBekenntnisse.The fourth chapter summarises the results of the analysis and presents Jessner’s Stil desWesentlichen, focussing on its key elements: the dramaturgical method, the role of scenicdesign, and of the actor and the relationship with the audience. The last section of the chapterinvestigates the correlation between Jessners’ Stil des Wesentlichen, Expressionism and NewObjectivity.
«ICH BIN EIN REGISSEUR DES WORTS UND NICHT DER DEKORATION»:L¿ESTETICA TEATRALE DI LEOPOLD JESSNER: QUASI UN ÓRGANON / S.v. D'orazio ; tutor: M. Castellari ; coordinatrice: M. V. Calvi. Dipartimento di Lingue e Letterature Straniere, 2021 Apr 09. 32. ciclo, Anno Accademico 2020. [10.13130/d-orazio-silvia-vincenza_phd2021-04-09].
LA SVOLTA AMOROSA DELLA FENOMENOLOGIA FRANCESE. LA POSSIBILITÀ DI UN'ETICA FENOMENOLOGIA
"The question of ethics is as old as phenomenology. Husserl held numerous series of lectures on ethics throughout his academic career. When Heidegger wrote Sein und Zeit, some critics thought the book was a text on moral philosophy and ethics. According to Husserl, the task of phenomenology with regard to ethics is the clarification of the formal conditions of practical reason. Phenomenology (in accordance with its own method) must suspend judgment on moral values and must focus on the formal connections necessary for ethical reasoning. However, analyzing the texts, it is clear how Husserl gradually accords more space to circumstance: the study of moral judgment shows how singularity and historicity cannot be underestimated. When man has to choose what is right, the most universal thing is the importance of the singular. As far as Husserl is concerned, the main driving force of ethical actions is love, and love can only be understood within circumstance. From this point of view, Heidegger and Husserl show unprecedented proximity. According to Heidegger, there is no difference between ethics and ontology: being is what happens.Ethics shows the potential and limits of phenomenology: the phenomenological investigation of ethics indicates the possibility of further avenues of investigation. The subject ""becomes himself"" in the exercise of ethical discernment. Levinas and Marion follow the path of the investigation of how the subject becomes such in relation to otherness. In their investigations they cross the question of love, which is fundamental for the constitution of the subject. The second part of the research evaluates the path of French phenomenology as an important theoretical attempt to investigate what Husserl only indicated by impacting, writing about the weight of singularity and history."
LA SVOLTA AMOROSA DELLA FENOMENOLOGIA FRANCESE. LA POSSIBILITÀ DI UN'ETICA FENOMENOLOGIA / M. Belli ; tutor: C. di Martino, L. Solignac ; supervisori: A. Caputo, E. Falque. Università degli Studi di Milano, 2022 Dec 12. 35. ciclo, Anno Accademico 2022.
THE MEDIATING EFFECT OF INCOME AND EDUCATION USING THE UK NATIONAL AND DIETARY SURVEY (NDNS) DATA
Diet substantially contributes to socioeconomic inequalities in cardiovascular morbidity and mortality. High adherence to the dietary approach to stop hypertension (DASH) has been proved effective in lowering blood pressure in patients with cardiovascular disease (CVD) as well as to prevent CVD risk factors in the general population. Little is known about time trends in diet quality and associated inequalities in the United Kingdom (UK). In addition, the causal pathway between education and dietary choices has not been fully explained and the role of income in preventing a healthy diet has not been clarified. This doctorate firstly, aimed to quantify the differences in adherence to the DASH in relation to socioeconomic position (SEP) in the UK and to evaluate recent trends. Secondly, it aimed to quantify the mediating effect of income on the relationship between education and the DASH score in the UK population.Data used for analysis was obtained from three waves of the National Diet and Nutrition Survey Rolling Programme (NDNS) 2008-2012, 2013-2014 and 2015-2016). The DASH score was calculated based on Fung et al methodology and was calculated using sex-specific quintiles of DASH items. For the first part of the analysis (Paper 2) data analysis included 6435 subjects aged 18 and older who participated in the NDNS. Multiple linear regressions were used to evaluate the relationship between the socioeconomic variables and the DASH score. Quantile regression analysis was used to model the median intake of each component as a function of the socioeconomic variable and the survey year. In the second analysis (Paper 3), analysis was done on 4864 subjects aged 18 and older. Counterfactual-based mediation analysis was carried out to decompose the total effect of education on DASH score into average direct effect (ADE) and average causal mediation effect (ACME) mediated by income.A gradient relationship between the DASH score and all socioeconomic variables emerged with increasing values of the score at higher socioeconomic positions (SEP effect p value: <0.0001 for education, occupation, and income) in the initial analysis. The interaction term between survey year and the socioeconomic variables was not significant showing that the trend was not different across socioeconomic groups (p >0.05). The estimated difference between people with no qualification and those having the highest level of education was -3.59 points (95% CI: -3.91; -3.20). The difference between people engaged in routine occupations and those engaged in high managerial and professional occupations was -3.40 points (95% CI: -3.87; -2.92), and the difference between subjects in the first fifth and last fifth of the household income distribution was -2.73 points (95% CI: -3.16; -2.29). The widest socioeconomic differences emerged for consumption of fruit, vegetables, whole grains, nuts, seeds and legumes. Mediation analysis indicated that the overall mediating effect of income on the relationship between education and the DASH score was only partial, with an estimated proportion mediated ranging between 6 to 9%. The mediating effect was higher among women (11.6%) and younger people (17.9%).Findings from this doctorate add an important contribution to the existing literature and more importantly, provide an updated picture of socio-economic inequalities in diet amongst UK adults in context of the whole diet. The results show that overall, the DASH score increased over time, yet the overall score remains low. Moreover, persistent disparities between individuals with higher versus lower SEP were observed. Additional analysis indicates that low income plays a modest role in explaining educational differences in the UK population. Further research is needed to investigate which other factors may explain differences in diet quality.In conclusion, findings in this doctorate have substantial implications for public nutrition policy. An immediat
THE MEDIATING EFFECT OF INCOME AND EDUCATION USING THE UK NATIONAL AND DIETARY SURVEY (NDNS) DATA / L. Patel ; tutor: C. LA VECCHIA. Dipartimento di Scienze Cliniche e di Comunità, 2021 Mar 10. 33. ciclo, Anno Accademico 2020. [10.13130/patel-linia_phd2021-03-10].
DESIGN AND IMPLEMENTATION OF DIGITAL MATHEMATICAL DISCUSSION: FOCUS ON STUDENTS¿ COLLABORATIVE PROCESSES AND THE DEVELOPMENT OF COLLECTIVE ALGEBRAIC THINKING
The integration of digital technologies into education provides significant opportunities to enhance mathematics teaching and learning, particularly by fostering collaboration and communication beyond traditional classroom settings. The research presented in this thesis contributes to the ongoing investigation on the role of digital technologies to support mathematical discussion, which is a didactic methodology designed to actively involve students in constructing mathematical knowledge through interaction with peers and experts. While existing research has already started addressing the support digital technologies can give to the design and unfolding of synchronous mathematical discussions, taking place in classrooms or through conferencing platforms, this thesis focuses on asynchronous mathematical discussions. In fact, asynchrony allows participants to engage with the discussion at their own pace, encouraging thoughtful reflections and giving time to process information. Written communication, in turn, encourages participants to make their reasoning more explicit, making their thinking visible. Research on asynchronous mathematical discussion remains an understudied emerging area, especially in high school contexts. Thus, we aim to contribute to this research gap introducing and investigating the design and implementation of Digital Mathematical Discussion.Digital Mathematical Discussion is a didactic methodology stemming from both mathematical and asynchronous discussions. It comprises three phases: group-work within chats to solve a mathematical problem; whole-class balance discussion within Padlet to compare solutions; and wrap-up classroom discussion. For the scopes of this research, we focus on the first two phases, as we intend to deepen the investigation on asynchronous mathematical discussions. We answer the questions of what kind of students’ collaborative processes are fostered in the digital environments facilitating a Digital Mathematical Discussion and how they intertwine with the development of collective algebraic thinking. To investigate the collaborative processes we refer to the dimension of social modes of co-construction, while for the use of algebra as a thinking tool we focus on conceptual frames and anticipating thoughts.To address the research questions, we adopt a design-based research methodology, including a pilot study with university students familiar with traditional mathematical discussion, and two cycles of experimentations with grade 9 classes. The tasks proposed within the experimentations highlight the use of algebra as a thinking tool and enable students to explore numerical situations, formulate conjectures, and construct proofs through the use of algebraic language.Our findings reveal different outcomes on the intertwining between social modes of co-construction and the development of collective algebraic thinking by students within the different digital environments facilitating Digital Mathematical Discussion. Within the chats, we identify varying collaboration processes, more or less effective in actively considering and building on each other’s proposals and for the development of a shared understanding and collective algebraic thinking. Within Padlet, there is a flattening of the social modes of co-construction and a prevalence of individual algebraic thinking. Moreover, the research provides insights for the design of a platform to facilitate Digital Mathematical Discussions, which have informed the development of a prototype.
Design and implementation of Digital Mathematical Discussion: focus on students' collaborative processes and the development of collective algebraic thinking / S. Gagliani Caputo ; tutor: L. Branchetti, A. Cusi, S. Mantovanicoordinatore: G. Ciraolo. Dipartimento di Matematica Federigo Enriques, 2025 May 19. 37. ciclo, Anno Accademico 2024/2025.
NOVEL INSIGHTS ON ALLAN-HERNDON-DUDLEY SYNDROME: TRANSCRIPTIONAL PROFILING AND FUNCTIONAL CHARACTERIZATION OF THREE GENETIC VARIANTS IN SLC16A2 GENE
Allan-Herndon-Dudley Syndrome (AHDS) is a rare genetic X-linked pediatric disorder, affecting essentially males, with intellectual and motor development impairment. It is caused by mutations in the SLC16A2 gene, which encodes for MCT8, a transporter responsible for thyroid hormone (TH) transport across cell membranes. THs are crucial for the development of several organs, mainly the brain and are involved in many physiological processes. Variants in SLC16A2 impact on the functionality and localization of MCT8 in various ways. The resulting clinical condition is characterized by brain hypothyroidism and thyrotoxicosis in peripheral tissues, leading to a range of symptoms, including cognitive delays, hypotonia, spasticity, and dystonia. In this study, we provide a deep investigation about the impact of three different pathological variants in SLC16A2 gene, identified in AHDS affected pediatric patients, on the pathogenetically mechanisms of AHDS. Particularly, from skin biopsies, we isolated primary fibroblasts from 3 AHDS patients and 2 healthy controls matched by sex and age to study the pathological readouts of the disease and identify novel related molecular pathways. With this purpose, we applied different techniques, including MTT and Live&Dead assays to characterize the obtained fibroblast cell lines. Moreover, we performed RT-qPCR, Western Blot, Immunofluorescence experiments, to investigate the functioning and the localization of MCT8 transporter in these cells. RNA-sequencing analysis provided us an overview about differentially expressed genes correlated to distinctive mechanisms of the disease, such as neuronal and glial specification deregulation; a reduced protein synthesis as a result of ribosome structural components’ impairment, a reduced mitochondrial activity and lipid droplets’ accumulation as a result of a disrupted thyroid hormone signaling. Overall, our study confirms the use of patients’ derived fibroblasts as a reliable model to study in vitro rare conditions, as well as showed us the importance of personalized medicine when patients carry different mutation related to the disease. Nevertheless, major limitations of the study are also exposed to represent the first starting point for overcoming them and finding a definitive cure for this debilitating condition.
NOVEL INSIGHTS ON ALLAN-HERNDON-DUDLEY SYNDROME: TRANSCRIPTIONAL PROFILING AND FUNCTIONAL CHARACTERIZATION OF THREE GENETIC VARIANTS IN SLC16A2 GENE / L. Esposito ; tutor: G. V. Zuccotti ; co-tutor: C. Cereda ; coordinator: F. Cheli. Dipartimento di Scienze Biomediche e Cliniche, 2025. 37. ciclo, Anno Accademico 2024/2025.
NORMAL FORM AND KAM METHODS FOR HIGHER DIMENSIONAL LINEAR PDES
"In this thesis an approach to linear PDEs on higher dimensional spatial domains is proposed. I prove two kinds of results: first I develop an algorithm which enables to obtain reducibility for linear PDEs which depend quasi-periodically on time, and I apply it to a quasilinear transport equation of the form∂_t u= ν•∇u+ ε P (ωt)u on the d-dimensional torus T^d, where ε is a small parameter, ν and ω are Diophantine vectors, P (ωt)=V(x,ωt)•∇+W(ωt), V is a smooth function on T^(d+n) and W(ωt) is an unbounded pseudo-differential operator of order strictly less than 1. The strategy is an extension of the methods originally developed in the context of quasilinear one dimensional equations. It consists in first using quantum normal form techniques in order to conjugate the original system to a new one with a smoothing perturbation, and then exploiting the smoothing nature of the new perturbation in order to balance the effects of the small denominators, which in this problem accumulate very fast to 0.The quantum normal form procedure developed in order to obtain reducibility for the above transport equation is global in phase space. In order to overcome such a limitation, the second problem I tackle in this thesis is that of developing a local quantum normal form procedure, which could be applied to much more general systems. As the simplest relevant model containing all the difficulties of the general case, I consider the operator H=-∆+V(x) with Floquet boundary conditions on the flat torus T^d_Γ, where T^d_Γ is the manifold obtained as quotient between the d-dimensional space R^d and an arbitrary d-dimensional lattice Γ, with the purpose of adapting the quantum normal form procedure to deal with this operator. As a result, I prove for the operator H a Structure Theorem à la Nekhoroshev, and I characterize the asymptotic behavior of all its eigenvalues. The asymptotic expansion is in |λ|^{-δ}, with δ ∊ (0, 1) for most of the eigenvalues λ (stable eigenvalues), while it is a ""directional expansion"" for the remaining eigenvalues (unstable eigenvalues)."
NORMAL FORM AND KAM METHODS FOR HIGHER DIMENSIONAL LINEAR PDES / B. Langella ; tutor: D. Bambusi ; co-tutor: R. Montalto ; coordinatore: V. Mastropietro. Dipartimento di Matematica Federigo Enriques, 2020 Dec 17. 33. ciclo, Anno Accademico 2020. [10.13130/langella-beatrice_phd2020-12-17].
PERIODIC AND QUASI-PERIODIC ORBITS IN NEARLY INTEGRABLE HAMILTONIAN SYSTEMS
"The study of periodic and quasi-periodic orbits in nearly integrable Hamiltonian systems is a long standing and challenging problem, that dates back to Poincaré. Quoting Poincaré, they represent ""the only opening through which we can try to enter a place which, up to now, was deemed inaccessible"". The aim of this thesis is to find effective and constructive algorithms for constructing both periodic and quasi-periodic solutions via a modification of the normal form methods related to Kolmogorov's theorem. The thesis is divided in two parts. The first part concerns the classical problem of the continuation of periodic orbits surviving to the breaking of invariant maximal or lower dimensional completely resonant tori in nearly integrable Hamiltonian systems: we here propose a new scheme which allows to deal with the problem of degeneracy at any order of perturbation. The second part regards the development of a variation of the Kolmogorov's normalization algorithm, by avoiding the so-called translation step at the price of fixing only the final frequency, while the initial one can only be determined a posteriori."
PERIODIC AND QUASI-PERIODIC ORBITS IN NEARLY INTEGRABLE HAMILTONIAN SYSTEMS / V. Danesi ; tutor: T. Penati ; co-tutor: M. Sansottera ; coordinatore: V. Mastropietro. Dipartimento di Matematica Federigo Enriques, 2021 Feb 22. 33. ciclo, Anno Accademico 2020. [10.13130/danesi-veronica_phd2021-02-22].
CHOICE AGENDAS IN DISABILITY POLICY AND PRACTICE: AN ANALYSIS THROUGH THE LENSES OF PROFESSIONAL ACTORS. THE CASES OF ENGLAND AND LOMBARDY IN COMPARISON
‘Choice’ is nowadays a popular concept in welfare policy. In the past twenty to thirty years, the popularity of choice has in fact spread from the private sphere of everyday life to the less-private context of public policy. Choice, however, has not entered public policy on its own. Different actors - from disability activists to politicians and policy-makers - have advocated for choice, often for diverse and contrasting purposes. Also, specific tools have been associated with the entry of choice in welfare services, and these tools are neither purely technical nor neutral. This thesis aims to understand the effects of ‘choice agendas’ at the point in which they make their way into public services; and it does so through the lenses of the professional actors in charge of translating ‘choice’ from policy into practice. In particular, drawing on street-level-bureaucracy, policy practice and network theories, choice agendas are analysed in the specific context of social care services for disabled people as developed both in England and the Italian region of Lombardy. The application of a mix of qualitative research methods brought me to the conclusion that power differences and asymmetries are at play among different professional actors; with some having stronger discretion and policy steering capacity compared to others. Such differences imply that only certain visions of choice are legitimated in practice, with questionable implications in terms of the nature of the transformation brought about by choice agendas and their contribution in offering more ‘choice and control’ to disabled people.
CHOICE AGENDAS IN DISABILITY POLICY AND PRACTICE: AN ANALYSIS THROUGH THE LENSES OF PROFESSIONAL ACTORS. THE CASES OF ENGLAND AND LOMBARDY IN COMPARISON / F. Pozzoli ; promotores: J. W. DUYVENDAK, F. BIOLCATI RINALDI. Dipartimento di Scienze Sociali e Politiche, 2021. 32. ciclo, Anno Accademico 2020.
OPTOMECHANICAL COLLECTIVE EFFECTS USING COLD ATOMS IN FREE SPACE: COLLECTIVE ATOMIC RECOIL LASING & OPTICAL BINDING
This theoretical doctoral thesis investigates the collective effects that emerge incold atomic systems caused by light-scattering in free space. Two specific cases areinvestigated: the collective atomic recoil laser (CARL) effect in a cold gas, withoutoptical cavity, and a novel cooperative cooling effect via optical binding (OB) withcold atoms.As a main objective, this theoretical project investigates the spatial gratingstructures and the backward radiation that appears in a cold atomic cloud whenit is irradiated by a single far-detuned laser beam, also known as CARL effect.While this effect has traditionally been described using a ring cavity, the study isperformed here in free space, in the absence of such a cavity. Both 2D and 3D cloudsshow a transition from single-atom isotropic scattering to collective directionalscattering. The effect is shown by the derivation and numerical solution of a set ofmulti-particle motion equations coupled by a self-consistent optical field, which isinspected with both a scalar model and a vectorial model. New original approachesare used to address the numerical study of the dynamics of the atomic system, suchas molecular dynamics (MD) algorithms.A second system emerged, from the attempt to understand the main objective,where a few atoms rearrange themselves into crystalline atomic structures, with aperiodicity between particles close to the optical wavelength. The atomic systemis initially confined into a 2D plane (or 1D string) using two (or four) counter-propagatinglaser beams. Due to the multiple scattering experienced by all theparticles in the system, a dipole-dipole force arises among them, generating a non-trivialdynamical trapping potential landscape that compels the atoms, to self-organizeat distances multiple of the light wavelength. When atoms are rearrangedinto an atomic crystal, the force acting on each particle depends on the position ofthe others, thus allowing to study the stability of such optically bound structures.In addition, it turns out that a non-conservative force is generated from thedipole-dipole interaction, allowing the system to be cooled by controlling the valueof certain parameters. This new phenomenon arises as a direct consequence of theuse of cold atoms instead of dielectric nanoparticles in an OB system. Therefore,besides the atomic external motion, internal degrees of freedom (DOF) of the atomsare considered by treating each atom as a dipole. This latter aspect is investigatedusing the coupled dipole equations. When multiple atoms are set in line, the coolingmechanism is collectively enhanced, generating a novel cooperative cooling effect.
OPTOMECHANICAL COLLECTIVE EFFECTS USING COLD ATOMS IN FREE SPACE: COLLECTIVE ATOMIC RECOIL LASING & OPTICAL BINDING / A. Tarramera Gisbert ; supervisore: N. Piovella ; direttore scuola dottorato: M.G. Paris. Dipartimento di Fisica Aldo Pontremoli, 2020 Dec 11. 32. ciclo, Anno Accademico 2019. [10.13130/tarramera-gisbert-angel_phd2020-12-11].
CORTICAL INHIBITION OF HYPERACTIVE ADAM10 PREVENTS CORTICO-STRIATAL CIRCUITRY DEFECTS IN HUNTINGTON¿S DISEASE
Huntington's disease (HD), a devastating neurodegenerative disorder caused by a CAG triplet expansion in exon 1 of the huntingtin gene (HTT), is characterized by progressive neuronal loss, predominantly in the striatum and cortex. Neuroimaging and electrophysiological studies consistently reveal that cortical projection neurons, the main presynaptic input to the striatum, are compromised early in the disease. While both cortical and striatal neurons are affected by mutant Htt, in vivo and in vitro studies have shown that selective mutant Htt expression specifically in cortical neurons is both necessary and sufficient to disrupt cortico-striatal circuitry integrity and drive striatal pathology. This highlights presynaptic cortical dysfunction as a critical contributor to striatal degeneration in HD, presenting an intriguing target for the development of novel drugs for HD. At the molecular level, the disruption of Brain- Derived Neurotrophic Factor (BDNF) synthesis and transport in cortical neurons is solidly associated with cortico-striatal circuitry dysfunction and striatal pathology in HD. As part of the continuing efforts to unravel the molecular mechanisms driving HD-related synaptic dysfunction, our group discovered A Disintegrin And Metalloproteinase Domain-containing protein 10 (ADAM10) as novel synaptic partner of Htt. ADAM10 exerts fundamental roles in the homeostasis of neuronal networks through its proteolytic activity on a plethora of synaptic adhesion proteins. In HD, the ADAM10/Htt complex is hampered, leading to ADAM10 accumulation at the synapse, which in turn disrupts synaptic function and contributes to cognitive decline in HD mice. More recent findings revealing ADAM10’s pivotal role in presynaptic homeostasis—specifically in the regulation of synaptic vesicle transport and release—have led us to propose the intriguing hypothesis that increased accumulation of active ADAM10 in HD cortical neurons may trigger cortico-striatal dysfunction and striatal pathology in HD. The aim of this PhD thesis is to investigate the contribution of presynaptic (cortical) ADAM10 in HD cortico-striatal circuitry dysfunction. To reconstruct the HD cortico-striatal circuitry I employed a microfluidic-based cellular platform with separated cortical and striatal compartments. The selective ADAM10 inhibitor GI254023X was administered independently to cortical and striatal neurons within the reconstructed network, and cortico-striatal connectivity was evaluated. Finally, to assess translational potential, we tested the therapeutic efficacy of intranasally delivered GI254023X in an HD mouse model.
CORTICAL INHIBITION OF HYPERACTIVE ADAM10 PREVENTS CORTICO-STRIATAL CIRCUITRY DEFECTS IN HUNTINGTON¿S DISEASE / A. Scolz ; scientific tutor: E. Cattaneo ; scientific tutor: C. Zuccato ; coordinator of the PhD school: S. Ricagno. Dipartimento di Bioscienze, 2025 Feb 17. 37. ciclo
RITI RELIGIOSI E PRATICHE DI SFRUTTAMENTO. IL CASO DEL SISTEMA CRIMINALE NIGERIANO IN ITALIA
"The thesis analyses the phenomenon of trafficking and the system of exploitation of prostitution involving Nigerian women in three distinct Italian contexts: Milan, Naples, and the Domizio coastal area (in the Campania region). The study examines the role played by the religious dimension in determining or not conditions of vulnerability. To this end, we tried to understand both the functions covered by the traditional rites, known as “juju rites”, practiced on women on behalf of the criminal organization by local priests (the native doctors), and the roles played by the pastors of the Nigerian Pentecostal churches present in many Italian cities. Gender is the second dimension of analysis taken into consideration, in relation to the criminal markets involved and their management, the determination or not of conditions of subordination-exploitation and the ability to emerge and recognize the status of ""victim of trafficking” by the institutions responsible for combating the phenomenon and protecting the actors (women but also men) directly involved.The basic hypothesis that drives the research is that it is not possible to understand the criminal phenomenon of the Nigerian trafficking and, more generally, the female-dominated criminal system that coordinates its management, if we do not try to understand (as far as possible) the religious dimension, that is the spiritual world on which most Nigerians seem to base their choices, whether they are inserted in a criminal or completely legitimate context."
RITI RELIGIOSI E PRATICHE DI SFRUTTAMENTO. IL CASO DEL SISTEMA CRIMINALE NIGERIANO IN ITALIA / F. Cabras ; tutor: M. Massari ; coordinatore: F. Basile. Dipartimento di Studi Internazionali, Giuridici e Storico - Politici, 2022 Jul 11. 34. ciclo, Anno Accademico 2021.
ECOLOGY OF HIGH-ALTITUDE ENVIRONMENTS: BIODIVERSITY, DYNAMICS, AND ANTHROPIC IMPACTS
A wide variety of organisms has been proven to thrive on the surface of glaciers and ice sheets, which have been proposed to be a proper biome. The glacial organisms described include a wide variety of taxa, which also show geographical variations in their distribution. Given the recency of this branch of research, a lot of questions still need to be answered, ranging from the taxonomical classification of many species detected on glaciers, to the biogeographical patterns of their distribution and the ecosystem services these organisms provide. This thesis is an attempt to contribute to the answer to some of these questions. We will first analyse biogeographical patterns in bacterial communities of cryoconite holes, finding that each glacier hosts a distinct bacterial community, even when glaciers are only a few hundred metres apart. This variation is higher than the seasonal and year-to-year variation of cryoconite holes bacterial communities that occurs on some glaciers and suggests a biogeographical pattern of distribution. Then, we will describe the trophic interactions that occur and shape the functioning of a model glacier ecosystem (the Forni Glacier), determining the importance of cyanobacteria and algae as primary producers, and of springtails and tardigrades as consumers. The following section is dedicated to contaminants, including sources of contamination (e.g., ski infrastructure) and their effects on microplastic contamination, latitudinal patterns in microplastic contamination at different sites, and metal and persistent organic contamination on the Alps. Our results show that ski infrastructure has a significant effect on microplastics concentrations, shape, and composition, and that latitude may be a factor in explaining the distribution of microplastics at a broader scale. In contrast, contamination by metals and persistent organic pollutants at Alpine scale seems to be mainly driven by local sources. The last section investigates the effects of pollutants of the glacier biota, by means of direct measurements of radioisotopes bioavailability and their uptake by glacier biota, and of a review of literature to assess the risk due to a broader number of contaminants.
ECOLOGY OF HIGH-ALTITUDE ENVIRONMENTS: BIODIVERSITY, DYNAMICS, AND ANTHROPIC IMPACTS / A. Crosta ; tutor: R. Ambrosini, M. Parolini ; coordinatore: M. Guarino. Dipartimento di Scienze e Politiche Ambientali, 2024 Dec 19. 37. ciclo, Anno Accademico 2023/2024.
TREATMENT OF ACUTE CORONARY SYNDROMES IN OLDER ADULTS
BackgroundDespite the increased proportion of elderly patients among those admitted for an acute coronary syndrome (ACS), elderly population has been underrepresented in prospective studies and randomized clinical trials. Nevertheless, older people are often frail, which means that they have low reserve capacity and decreased ability to manage complex physiologic stress, with a significant multimorbidity burden. Accordingly, they deserve dedicated studies to better improve clinical decision making in daily practice.AimTo provide additional information on risk factors and prognosis of elderly patients admitted to hospital for an ACS. MethodsThe project was based on three post-hoc analyses of the Elderly ACS 2 Randomized Trial- NCT01777503. The trial included ACS patients aged75 years and aimed to compare clopidogrel versus prasugrel 5 mg on top of acetyl salicylic acid for long term secondary prevention and bleeding events. Three main research questions (RQ) were addressed: 1) the role of a specific inflammatory disease (psoriasis) as risk factor for ACS, evaluated within a case-control study; 2) the role of ST-segment elevation myocardial infarction (STEMI) as presenting ACS type as risk factor for cardiovascular, non-cardiovascular death and stroke, evaluated through a cohort study with competing risk analysis; 3) the role of the residual angiographic burden (after percutaneous coronary intervention-PCI) in predicting 1-year mortality and cardiovascular events, evaluated through the change in net benefit (NB) over a core prediction model including the most relevant clinical variables and basal angiographic burden.ResultsRQ1: The prevalence of psoriasis was lower among cases (12/1455, 0.8%) than among controls (18/1108, 1.6%). The multivariate OR of ACS according to history of psoriasis (adjusted for age, sex and smoking) was 0.51 (95% confidence interval: 0.23–1.09). RQ2: Patients with STEMI had a higher risk of cardiovascular death (cause-specific hazard ratio, cHR 1.85; 95% confidence interval, CI: 1.02-3.36), non-cardiovascular death (cHR 2.10; 95% CI, 1.01-4.38), and stroke (cHR 4.8; 95% CI, 1.7-13.7) as compared to patients with NSTEMI. RQ3: The inclusion of angiographic residual burden gave little incremental value in the standardized NB compared to the core model.ConclusionsOur data does not support an association between psoriasis and risk of ACS in the elderly. In these patients, STEMI is an important predictor of cardiovascular death, non-cardiovascular death and stroke. The residual angiographic burden does not improve 1-year prediction of adverse outcome compared with a model including clinical variables and the basal angiographic burden. Elderly patients are a heterogenous, complex, and high- risk group whose management requires a multidimensional clinical approach beyond coronary anatomic variables.
TREATMENT OF ACUTE CORONARY SYNDROMES IN OLDER ADULTS / N. Morici ; tutor: G. Alicandro, C. V. B. La Vecchia ; direttore del dottorato: C. V. B. La Vecchia. Dipartimento di Scienze Cliniche e di Comunità, 2021 Mar 10. 33. ciclo, Anno Accademico 2020. [10.13130/morici-nuccia_phd2021-03-10].
HIGH PERFORMANCE COMPUTING MACHINE LEARNING METHODS FOR PRECISION MEDICINE
"Precision Medicine is a new paradigm which is reshaping several aspects of clinical practice, representing a major departure from the ""one size fits all"" approach in diagnosis and prevention featured in classical medicine. Its main goal is to find personalized prevention measures and treatments, on the basis of the personal history, lifestyle and specific genetic factors of each individual.Three factors contributed to the rapid rise of Precision Medicine approaches: the ability to quickly and cheaply generate a vast amount of biological and omics data, mainly thanks to Next-Generation Sequencing; the ability to efficiently access this vast amount of data, under the Big Data paradigm; the ability to automatically extract relevant information from data, thanks to innovative and highly sophisticated data processing analytical techniques. Machine Learning in recent years revolutionized data analysis and predictive inference, influencing almost every field of research.Moreover, high-throughput bio-technologies posed additional challenges to effectively manage and process Big Data in Medicine, requiring novel specialized Machine Learning methods and High Performance Computing techniques well-tailored to process and extract knowledge from big bio-medical data.In this thesis we present three High Performance Computing Machine Learning techniques that have been designed and developed for tackling three fundamental and still open questions in the context of Precision and Genomic Medicine: i) identification of pathogenic and deleterious genomic variants among the ""sea"" of neutral variants in the non-coding regions of the DNA; ii) detection of the activity of regulatory regions across different cell lines and tissues; iii) automatic protein function prediction and drug repurposing in the context of biomolecular networks. For the first problem we developed parSMURF, a novel hyper-ensemble method able to deal with the huge data imbalance that characterizes the detection of pathogenic variants in the non-coding regulatory regions of the human genome. We implemented this approach with highly parallel computational techniques using supercomputing resources at CINECA (Marconi – KNL) and HPC Center Stuttgart (HLRS Apollo HAWK), obtaining state-of-the-art results.For the second problem we developed Deep Feed Forward and Deep Convolutional Neural Networks to respectively process epigenetic and DNA sequence data to detect active promoters and enhancers in specific tissues at genome-wide level using GPU devices to parallelize the computation.Finally we developed scalable semi-supervised graph-based Machine Learning algorithms based on parametrized Hopfield Networks to process in parallel using GPU devices large biological graphs, using a parallel coloring method that improves the classical Luby greedy algorithm.We also present ongoing extensions of parSMURF, very recently awarded by the Partnership for Advance in Computing in Europe (PRACE) consortium to further develop the algorithm, apply them to huge genomic data and embed its results into Genomiser, a state-of-the-art computational tool for the detection of pathogenic variants associated with Mendelian genetic diseases, in the context of an international collaboration with the Jackson Lab for Genomic Medicine."
HIGH PERFORMANCE COMPUTING MACHINE LEARNING METHODS FOR PRECISION MEDICINE / A. Petrini ; tutor: G. Valentini, Giorgio, G. Grossi ; coordinatore corso di dottorato: P. Boldi. Dipartimento di Informatica Giovanni Degli Antoni, 2021 Mar 22. 33. ciclo, Anno Accademico 2020.
GREEN MINDS: KNOWLEDGE-BASED STRATEGIES FOR THE ADOPTION OF A PLANT-BASED DIET IN 3-6 Y.O. CHILDREN
This dissertation deals with how to design new knowledge-based educational programs that can help young (3- to 6-year-old) children in overcoming food neophobia, which is a disposition to reject unfamiliar food even before tasting it (Dovey et al. 2008) and constitutes a major hindrance to the adoption of healthy and sustainable eating habits during the developmental phase in which food preferences first take shape. Recent studies have in fact found correlations between poorer performance in cognitive tasks tapping into conceptual food knowledge (e.g., food categorization and property inference) and higher scores on the Child Food Rejection Scale (Rioux et al., 2017) among preschool children (Lafraire et al., 2016; Rioux et al., 2018; Pickard et al., 2021; Foinant et al., 2022), thus suggesting that children’s food concepts play an important role in determining their food behavior. Anyways, the educational potential of this connection is still unaddressed in the literature.The General Introduction outlining the general topics of this work, some key terms, and the methodological approach I’ve attended throughout this research project is followed by three research papers, which constitute the bulk of the dissertation. The first one is a literature review, whose aim is that of clarifying the notion of “conceptual food knowledge” in the light of extant experimental research on young children’s food cognition. Conceptual food knowledge in this population can be unpacked into a more fine-grained typology encompassing three main subtypes of knowledge that most children proved capable of mastering by the sixth year of life: taxonomic knowledge, which hinges on set-inclusion relations among food categories, relations knowledge, which involves contextual thematic and script relationships between food items and events, and value-laden knowledge, which encompasses evaluative and ad-hoc food knowledge. This typology is the first one of this kind in the literature and highlights the richness of young children food knowledge. Specifically, the review suggests that conceptual flexibility and knowledge about conceptual relations are key factors to build a mature understanding of the food domain.The fact that young children can attend various types of conceptual food knowledge raises questions about how to conceive of the “knowledge gaps” displayed by neophobic children. In fact, poorer performance in experimental tasks could be due to different reasons, such as children lacking specific types of conceptual knowledge, or them not being flexible enough to access the right type of knowledge for the task at hand, or again the fact that information conveyed by their conceptual representations and by the experimenters’ differ in content. Part two thus takes a step back from early food neophobia and food education to address the broader issue of how to study conceptual differences in the food domain, which is a crucial to the understanding of behaviors and interactions that call for interindividual coordination (e.g., communication). By introducing the notion of conceptual mismatch, I propose an original approach which integrates philosophical insights on food conceptual pluralism and experimental research practice in cognitive science: I first define “conceptual mismatch” as the state that obtains between two conceptual representations that with respect to the concepts they embed or the relationships between such concepts, and then proceed to single out different types of conceptual mismatches that have been investigated in the food domain. Extensional mismatches occur when two conceptual representations refer to distinct categories, intensional mismatches encompass scenarios in which the modes of presentation of two different conceptual representations map onto the same category, and ontological mismatches capture differences involving the conceptual relations that structure two food ontologies.
GREEN MINDS: KNOWLEDGE-BASED STRATEGIES FOR THE ADOPTION OF A PLANT-BASED DIET IN 3-6 Y.O. CHILDREN / M. Gandolini ; supervisors: A. Borghini, J. Lafraire : phd coordinators: F. Guala, C. Caracaillet. - Milano. Dipartimento di Filosofia Piero Martinetti, 2025 Jun 09. 37. ciclo, Anno Accademico 2023/2024.
PRIVILEGE LOST: THE COUNTRY HOUSE, ARISTOCRACY AND ENGLISHNESS IN NANCY MITFORD'S NOVELS
"This thesis explores the connections between Englishness and the English aristocracy in Nancy Mitford’s novels through a detailed analysis of their symbolic-cultural representation: the English country house. The main purpose of the present work, which combines literary, biographical, historical, geographical, architectural and social analysis, is the re-evaluation of Mitford’s undervalued literary production, which has been relegated to the category of conservative mass entertainment – also due to the thin veil of humour, satire, irony and sarcasm that characterises it – and its claims to cultural and historical relevance in interwar British fiction. Mitford's biography clearly influenced her narrative choices in terms of characters, settings and themes: her upper-class background, that of an English noblewoman who describes in detail, carefully examines and, at the same time, subtly lampoons her own social class, constructing her fictional characters as the perfect caricatures of aristocratic types, has been taken into consideration in the analysis of her novels since it not only lies at the basis of her literary output, but it also constitutes a unicum among the interwar English writers of country-house novels. The present thesis is structured in two main parts. The first introductory one pinpoints a few methodological directions, a specifically angled recognition of the studies that have so far dealt with Englishness, the myth of the English countryside and country house-fiction, and offer a detailed examination of the historical background to the decline of English country houses and of the landed classes in Britain in the interwar period. The second section features close reading and close observation, with the aim to explore the connections between the country house, aristocracy and Englishness in Mitford’s novels. Such connections characterise almost all her works, and trigger reflection on the reasons why and on the ways in which they are implicated in the narration: the declining English country house as a former repository of essential Englishness, the interwar English countryside still considered by the old aristocracy as a symbol of traditional values and as a bucolic refuge from the chaos of the modern world, the opposition between the declining Victorian and Edwardian nobility that hides behind the curtains of its country mansions, unwilling to accept the loss of its former power and prestige and the young, modern aristocrats and members of the Bright Young People of the 1920s and ’30s, who enjoy the pleasures of life between London and rural England, seen by them only as “commodified” landscape ready to be consumed, but which they are generally unable to fit into. Albeit a crude version of her more mature ones, her interwar novels in the strict sense, which are the object of analysis in chapter one and two, already reveal many similarities in the rural settings – a castle in the Scottish Highlands in ""Highland Fling"" (1931), small cottages and grand country houses in the Cotswolds both in ""Christmas Pudding"" (1932) and in ""Wigs on the Green"" (1935) –, and in the upper- and upper-middle-class characters (members of the old British nobility and of the Bright Young People). Mitford’s most famous novels, ""The Pursuit of Love"" (1945) and its companion volume, ""Love in a Cold Climate"" (1949), which were written during and after World War II, mainly focus “on the powerful cultural image of the English country house”, celebrating on the surface “a conservative, largely upper-class view that appealed to a nation devastated by war and to later generations holding fast to a particular notion of Englishness amid great historical and social change”, but in fact concealing “a sharply critical study of a declining upper class, incapable of maintaining its traditional feudal role as legal and moral authority” (Kawano 2017: 13). The fourth chapter, which takes into account Mitfo"
PRIVILEGE LOST: THE COUNTRY HOUSE, ARISTOCRACY AND ENGLISHNESS IN NANCY MITFORD'S NOVELS / F. Prina ; tutor: N. Brazzelli ; coordinator: M.V. Calvi. Dipartimento di Lingue, Letterature, Culture e Mediazioni, 2023 May 05. 35. ciclo
PROBABILISTIC MODEL CHECKING WITH MARKOV MODELS SEMANTICS: NEW DEVELOPMENTS AND APPLICATIONS
Contemporary society is increasingly dependent on the use of autonomous computational systems. Being sure that these systems behave appropriately and do what they have been designed for is thus fundamental. Several techniques have been developed to this purpose. Among them, model checking includes a series of methods that make use of computa- tional logic and automatic procedures to check whether systems’ behaviour satisfies given desirable properties.In recent years, due to the increasing prevalence of stochastic models, conventional model checking techniques have proven inadequate. The consequence has been the devel- opment of probabilistic model checking, a new research program aimed at developing proper tools to check the behaviour of stochastic systems. Among others, the probabilistic model checking with Markov models semantics and its related languages (i.e., the Probabilistic Computation Tree Logic (PCTL) and its extensions) has been particularly successful and is nowadays applied throughout a variety of different systems and domains. Nevertheless, there remain numerous unexplored developments and applications in this field. In this doctoral dissertation, we specifically concentrate on three such aspects that hold signifi- cant relevance to contemporary AI research. To each of these aspect, we will dedicate a chapter. The dissertation is structured as follows.In the first chapter, we present the essential foundational information regarding prob- ability theory and Markov models. Initially, we introduce fundamental terminology and refresh the reader’s memory on the significant philosophical interpretations of probability, the Kolmogorov axiomatization, and the associated calculus. Subsequently, we delve into the application of probability in studying stochastic systems. Our primary emphasis is on Markov models, including discrete-time Markov chains, Markov reward models, and Markov decision processes. These models are indeed considered the reference formalism for representing stochastic systems in the domain of model checking.In the second chapter, we provide an overview of the state of the art of model check- ing, with a particular emphasis on both single-agent and multi-agents probabilistic model checking. In the opening section, we discuss the fundamental ideas of the program verifica- tion research field, of which model checking represents one of the contemporary evolutions.We focus in particular on the philosophical debate that emerged in the 1980s between 34advocates and opponents of formal methods in computer science. This debate can be considered a pivotal milestone in the shift from traditional program verification methods to modern techniques such as model checking.In the subsequent sections, we then introduce various formalisms utilized in model checking for representing computational systems, specifying their attributes, and verifying their behaviors. For standard (non probabilistic) model checking, these formalisms include transition systems and their related logical languages (i.e., LTL and CTL), while for prob- abilistic model checking they include Markov models and the related language PCTL. We conclude the chapter by examining some recent extensions of probabilistic model checking suitable for the analysis of epistemic and probabilistic properties of stochastic multi-agent systems. Notably, the formalisms we consider include the logics CTLK [113], PCTLK [175], and COGWED [28], where the latter two formalisms will serve as the foundation for subsequent advancements and applications discussed in the remaining chapters of the dissertation.In the third chapter, we explore a potential connection between probabilistic model- checking and eXplainable AI (XAI), a recently-born field of research aimed at developing methods and tools to make opaque machine learning systems more humanly understand- able. The chapter is divided in two main parts. In t
PROBABILISTIC MODEL CHECKING WITH MARKOV MODELS SEMANTICS: NEW DEVELOPMENTS AND APPLICATIONS / A. Termine ; tutor: G. Primiero ; coordinatore: A. Pinotti. Università degli Studi di Milano, 2023 Jul 11. 35. ciclo, Anno Accademico 2022.
THE MATERIALITY OF KNOWLEDGE: THE COLONIAL ROOTS OF THE KNOWLEDGE ECONOMY
The present work aims to draw on the theoretical legacy of postcolonial critique, which includes the assumption of a fundamental continuity between colonial history and our postcolonial world, in order to rethink the current configuration of the relationship between capital and knowledge (usually analysed in terms of “knowledge society”, “knowledge economy”, and so on), in light of and in continuity with the history and structures of colonial capitalism. For this purpose, in the first place, the main neoliberal and critical theories concerning this relationship are discussed with a focus on the histories and temporalities they assume or entail, also including human capital theories and their connections with developmentalism, eugenics, and scientific racism. In the second place, a materialist conception of knowledge, of its relations with human labour and capital, and of its functions within capitalist production and reproduction is proposed in order to re-read its role in the history of capitalism during the long colonial modernity and to analyse the historical construction of Eurocentric modern knowledge. In the third place, the conclusions thus reached are used to “displace” the previously discussed theories, histories, and temporalities through the examination of significant cases from colonial history, mainly but not exclusively taken from early modern New Spain.
THE MATERIALITY OF KNOWLEDGE: THE COLONIAL ROOTS OF THE KNOWLEDGE ECONOMY / B. Paci ; tutor: G. Mormino ; coordinatore: A. Pinotti. Dipartimento di Filosofia Piero Martinetti, 2023 Apr 12. 35. ciclo, Anno Accademico 2022.
CHARACTERIZATION OF REM GENES INVOLVED IN THE REPRODUCTIVE DEVELOPMENT OF ARABIDOPSIS THALIANA
Role of REM34, REM35 and REM36 in gametophytes development.The REproductive Meristem (REM) gene family encodes for transcription factors belongingto the B3 DNA binding domain superfamily. In Arabidopsis thaliana, this family is composedof 45 members, preferentially expressed during flower, ovule, and seed developments. Onlya few members of this family have been functionally characterized: VERNALIZATION1(VRN1) and, most recently, TARGET OF FLC AND SVP1 (TFS1), which are involved in theregulation of flowering time and VERDANDI (VDD) which, together with VALKYRIE (VAL)controls the death of the synergid cell in the female gametophyte. We investigated the roleof REM34, REM35, and REM36, three closely related and linked on chromosome 4 geneswith similar expression profiles during the reproductive development of Arabidopsis.The functional characterization of REM34, REM35 and REM36 presented severalchallenges, as these genes are in linkage in the genome and redundant. We thus developedtwo different approaches, the first based on a multiplexed RNA interference (RNAi)technology and the second on the fusion of REM34 with the EAR repressive domain, tomodify their expression and identifying their role during inflorescence development.Plants generated with both methods were characterized by a partial sterility. An in situhybridization analysis showed that REM34, REM35 and REM36 share the same expressionpattern throughout reproductive organs development. As these genes were found to beexpressed both in female and male reproductive tissues, the developmental steps of bothgametes were followed in both the REM_RNAi and 35S:REM34-EAR lines; these analysesclearly showed that the partial sterility of these lines is caused by a postmeiotic block in bothfemale and male gametophytes. Furthermore, protein interaction assays revealed thatREM34 and REM35 interact, which suggests that they work together during the first stagesof gametogenesis.
CHARACTERIZATION OF REM GENES INVOLVED IN THE REPRODUCTIVE DEVELOPMENT OF ARABIDOPSIS THALIANA / F. Caselli ; tutor: V. Gregis ; scientific supervisor: M. Kater. Dipartimento di Bioscienze, 2021 Apr 14. 33. ciclo, Anno Accademico 2020.
MULTIVARIATE FLEXIBLE METAL ORGANIC FRAMEWORKS: THE ROLE OF FUNCTIONALIZED LINKERS, HETEROGENEITY AND DEFECTS IN ADSORPTION PROCESSES
Metal-Organic Frameworks (MOFs) are a class of synthetic porous crystalline materials based on metal ions connected through spacing ligands. They possess interesting properties such as, high porosity, high concentration of metal centers, flexibility, etc., which are difficult to find in conventional porous materials. As a result of these unique features, MOFs can maintain porous structures upon removal, inclusion, exchange, or reaction of a wide selection of molecular guests, making them useful for applications in selective gas adsorption/separation. Furthermore, a number of MOFs have the ability to respond to specific stimuli such as host-guest interactions, temperature or pressure change, light, etc. To date, most of the applications of these flexible MOFs is related to gas-storage and selective adsorption/separation, but it was soon recognized that the responsive properties of these materials have enormous potential in many areas, a really interesting example are NPs@MOFs composites which can be used to design novel “smart” heterogeneous catalysts capable of reversibly adapt to substrates and/or respond to regulating stimuli (e.g. reversible structural transitions upon adsorption of guest molecules). The work within this thesis is divided into two parts. The first one focuses on the understanding of the host-guest interactions during adsorption processes in highly stable, rigid, porous metal-organic frameworks. of formula Fe2(BDP)3x(BDP-NH2)3(1-x) (x = 0, 0.5, 1) with H2BDP-NH2 = 2-amino[1,4-bis(1Hpyrazol- 4-yl)benzene]. Their investigation was performed through a multi-technique approach exploiting high-resolution powder X-ray diffraction (HR-PXRD), advanced gas adsorption measurements and Monte Carlo simulations. In particular, the host-guest interactions during CO2 adsorption were studied via in situ HR-PXRD dosing CO2 in certain pressure ranges at different temperatures and compared between functionalized and bare systems. Rietveld refinements of the diffraction data collected at each pressure point was used to understand the preferential adsorption sites occupied by the gaseous probe and the host–guest interactions, as well as the breathing behaviour, whose mechanism was not yet completely described in the literature. The metal ion and ligand functionalization modulate the sorption properties of these materials, enhancing their affinity to CO2 molecules. Finally, new flexible, multivariate MOFs of general formula Zn(BDP)0.5(BDP-X)0.5 (X = NH2 or NO2) were synthesized. The deep characterization of the adsorption properties and the structural features of these materials will be crucial to understand the relation between the structural flexibility, functionalization degree and crystal size, with the gas adsorption and separation properties. The second part of this thesis describes a family of pillar-layered MOFs of general formula [M2(fubdc) 2P]n with M2+ = Zn2+, fu-bdc2- = diversely functionalised 1,4-benzenedicarboxylate) and how catalytically inert nanoparticles (silica NPs, 30 nm) can be embedded into the microporous ordered MOFs structures, obtaining hybrid materials where the NPs represent typically a meso-scale “defect” or “heterogeneity” to the system. In particular, the work was focused on the comparison between the plain MOFs and the new composite materials NPs@MOF. The parent [Zn2(bdc)2(dabco)]n is only weakly flexible, whereas the substituted frameworks [Zn2(fu-bdc)2(dabco)]n (with fu = DM, 2,5-bis(methoxy)- 1,4-benzenedicarboxylate or BME, 2,5-bis(2-methoxyethoxy)-1,4-benzenedicarboxylate) contract significantly upon guest removal to a narrow pore (np) form and expand again (lp form) upon adsorption of DMF, EtOH, or CO2, etc. In contrast, N2 is hardly adsorbed and does not open the narrow pore form. These “breathing” dynamics are attributed to the dangling side chains present on the linkers, which interact with mobile guest molecules as well as with themselves
MULTIVARIATE FLEXIBLE METAL ORGANIC FRAMEWORKS: THE ROLE OF FUNCTIONALIZED LINKERS, HETEROGENEITY AND DEFECTS IN ADSORPTION PROCESSES / S. Terruzzi ; tutor: V. Colombo ; co-tutor: A. Sironi ; coordinatore corso dottorato: D. Roberto. Dipartimento di Chimica, 2022 Jul 15. 34. ciclo, Anno Accademico 2021.
DISSECTING THE ROLE OF ACID SPHINGOMYELINASE INHIBITION IN GAUCHER DISEASE AND GBA-DEPENDENT PARKINSON¿S DISEASE IN VITRO MODELS
GBA gene encodes for the lysosomal enzyme β-glucocerebrosidase (GCase), which is involved in the catabolism of glucosylceramide (GlcCer) into glucose and ceramide. Biallelic mutations in GBA gene result in the GCase loss of function, leading to the accumulation of GlcCer and causing Gaucher Disease (GD). GD is a lysosomal storage disorder that manifests in various forms, ranging from the severe neuropathic type II to the less severe non-neurological type I.Interestingly, monoallelic mutations in GBA are now recognized as a major recessive genetic risk factor for Parkinson's Disease (PD), known as GBA-dependent PD. PD is a progressive neurodegenerative disorder characterized by the gradual loss of dopaminergic neurons and by the presence of aggregates of α-synuclein in the substancia nigra pars compacta of the brain. Up to now, the molecular mechanism linking GCase deficiency to the onset of neurodegeneration is still unknown. This is largely due to the lack of suitable in vitro models that accurately replicate the complexities of the human pathology associated with these conditions.To increase the complexity, it is important to consider that not all the individuals with GBA mutations develop PD. To understand this variability, the scientific community has explored the presence of other genetic variants in lysosomal enzymes that could influence the penetrance of GBA mutations.Our collaborators at Humanitas university have analysed a panel of 50 lysosomal genes, involved in the onset of lysosomal storage disorders when mutated, in a cohort composed by GBA asymptomatic carriers and GBA-mutated PD patients. They found that SMPD1 gene, coding for the enzyme acid sphingomyelinase (ASM), was more frequently mutated in controls than in GBA-PD, suggesting a possible protective effect of SMPD1 mutation in GBA carriers. In addition, a recent study has found that in a zebrafish GD model the inhibition of ASM, ameliorates the pathological phenotype and the survival, rescuing the mitochondrial chain function. Based on these considerations, the aim of my PhD project was to investigate whether the inhibition of ASM could have a beneficial role in GBA-related pathologies exploiting 2D and 3D in vitro neuronal models. To conduct this research, I generated two experimental models: a pharmacological one, using hiPSCs-derived dopaminergic neurons (hiPSCs-DANs), and a patient-derived one, using human midbrain organoids (hMOs). For the pharmacological model, I treated DANs for 30 days with conduritol-B- epoxide (CBE), a specific inhibitor of GCase, to induce the GD-PD phenotype. This is an experimental model that we have already demonstrated to show GCase deficiency, neurodegeneration and GlcCer accumulation. For the patient-derived one, I generate hMOs from iPSCs of a GD type I patient carrying the biallelic mutation N370S/F213I in GBA gene, who developed parkinsonism. I characterized this model and I demonstrated that it recapitulates the pathological features observed in the 2D model, with the addition of α-synuclein aggregation. These results point out that both dopaminergic neurons and organoids partially recapitulate the pathological phenotype observed in vivo. Thus, they constitute a good model for further investigations related to GD-PD pathology. Furthermore, I treated both models with amitriptyline (AMI), a functional inhibitor of ASM, to verify the effect of ASM inhibition in GD-PD pathology. I treated DANs 24h and 7 days before the collection at day 60 of differentiation. In hMOs instead the administration was longer, lasting 30 days and 70 days. Surprisingly, I found opposite results in the two models. Indeed, the acute (24h) and chronic (7 days) treatment with AMI has ameliorated the neurodegenerative phenotype in CBE-treated DANs, reducing GlcCer accumulation by increasing the non-lysosomal glucosylceramidase (NLGase) activity. Conversely, in hMOs the
DISSECTING THE ROLE OF ACID SPHINGOMYELINASE INHIBITION IN GAUCHER DISEASE AND GBA-DEPENDENT PARKINSON'S DISEASE IN VITRO MODELS / S. Breviario ; tutor: M. Aureli ; co-tutor: A. Di Fonzo ; coordinatore: C. Sforza. Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, 2024. 36. ciclo, Anno Accademico 2022/2023.
INVESTIGATING THE ROLE OF NOVEL GENES IN NEURODEVELOPMENTAL DISORDERS
Neurodevelopmental disorders (NDDs) are complex and diverse disorders of the developing human brain. Among them, intellectual disability (ID) is a lifelong pathological condition characterized by significant subaverage intellectual/cognitive functioning as well as adaptive functioning due to abnormalities of brain structure and function, whose onset occur before the age of 18. It affects 1–3% of the worldwide population, and no pharmacological therapies are currently available. ID is very heterogeneous in its etiology, being associated with a significant genetic and clinical variability. Some causes of ID have been defined, including genetic disorders, traumatic injuries, and prenatal events such as maternal infection or exposure to alcohol. Despite the common phenotype, the genetic bases are highly heterogeneous and apparently unrelated, and causes of ID remain unknown for up to 60% of all cases. Since the variety of ID makes challenging genetic and clinical diagnosis, in recent years a combined approach of exome sequencing and bioinformatics analysis has identified new ways to understand the causes of ID and help to interpret novel ID-causing genes. The identification of novel genes implicated in NDDs is fundamental to ameliorate diagnosis and treatment. Here, we have combined exome sequencing together with in silico, in vitro and in vivo experimental models to identify novel NDDs causative genes. First, we found out a novel heterozygous variant in Semaphorin 3E (SEMA3E) gene in a patient affected by severe ID and cognitive regression. We demonstrated that the identified mutation is causative of a complete loss of function of the Sema3E protein. Therefore, our in vitro and in vivo studies in mice might suggest that the mutation would possibly be involved in brain development and would explain, at least in part, ID pathogenesis. Further studies are needed to better understand the role of this family of genes in the susceptibility to NDDs, and the identification of additional mutations in patients affected by similar disorders will allow us to analyze the clinical relevance of SEMA3E.Moreover, we identified two brothers affected by an unrecognized genetic syndrome, comprising ID, and facial dysmorphisms, carrying mutation in Rhotekin (RTKN) and Natural Killer cell Triggering Receptor (NKTR). Our experiments have been important to reveal a novel role for two not well characterized genes in brain and head development. In particular, we demonstrated that RTKN has a central role in controlling cytoskeletal dynamics via Cofilin, by remodeling actin filaments. Besides, we elucidated a further role of NKTR as a novel RNA splicing protein downstream of SOX9, which is essential for craniofacial development and causing, if mutated, a novel craniofacial spliceosomopathy/neurocristopathy. The wealth of information deriving from this project will significantly increase our knowledge on RTKN and NKTR physiological role, which has been hitherto largely undescribed, and will validate these genes as novel causative gene of developmental syndromes. This project also provided evidence that the combined access to mutational screenings of patients with the application of in silico, in vitro and in vivo experimental models is effective in the discovery of novel genes implicated in complex disorders such as NDDs. Such information will be vital to ameliorate diagnosis and therapeutic intervention of disorders still largely idiopathic and being recognized as a leading cause of morbidity in children, causing great suffering for patients and their families and large costs for society.
INVESTIGATING THE ROLE OF NOVEL GENES IN NEURODEVELOPMENTAL DISORDERS / A.j.j. Paganoni ; tutor: A. M, Cariboni ; coordinatore: G. D. Norata ;. Dipartimento di Bioscienze, 2023 Apr 05. 35. ciclo, Anno Accademico 2022.
AN IMPROVED GENOMIC SURVEILLANCE APPROACH TO DISSECT THE SARS-COV-2 PANDEMIC
Genomic surveillance of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the only approach to rapidly monitor and tackle emerging variants of concern (VOC) of the COVID-19 pandemic. Such scrutiny is crucial to limit the spread of VOC that might escape the immune protection conferred by vaccination strategies. It is also becoming clear now that efficient genomic surveillance would require monitoring the host gene expression to identify prognostic biomarkers of efficacy and disease progression. Here we applied an integrated workflow for RNA extracted from nasal swabs to obtain in parallel the entire genome of SARS-CoV-2 and host respiratory epithelium transcriptome, representing the majority of Italian processed genomic samples. In addition, we have matured and applied novel proof-of-principle approaches to prioritize possible gain-of-function mutations by leveraging patients' metadata and isolated patient-specific signatures of SARS-CoV-2 infection. The goals mentioned above have all been achieved in a cost-effective manner that does not require automation, in an effort to allow any lab with a benchtop sequencer and a limited budget to perform integrated genomic surveillance on premises.
AN IMPROVED GENOMIC SURVEILLANCE APPROACH TO DISSECT THE SARS-COV-2 PANDEMIC / A. Grimaldi ; tutor: D. Cacchiarelli ; internal advisor: D. Di Bernardo ; external Advisor: N. Elvassore. Università degli Studi di Milano, 2022. 34. ciclo, Anno Accademico 2022.
(RE)IMPARARE DALLA PANDEMIA: HACER COMUNIDAD DI FRONTE AL COVID-19 (OAXACA, MESSICO)
This PhD thesis focuses on the social transformations triggered by the Covid-19 pandemic in the Mexican state of Oaxaca and, particularly, in the village of San Pablo Huitzo. The thesis combines nine months of ethnographic fieldwork with anthropological theoretical reflections. The social transformations examined represent a model of development beyond development. The interpretation of these transformations goes through the processes of learning, re-learning and re-signification of community knowledge and skills. These processes have been required to cope with Covid-19 and the structural limits of the health and public welfare system that the pandemic circumstance has highlighted. Many villages of Oaxaca adopted several strategies to prevent and treat viral contagion, according to their level of political, alimentary, medical, cultural, and organizational autonomy. These strategies were based on encierro (the closure of village’s borders), on a massive return to the earth, with a marked agroecological connotation, and the revitalization of traditional medicine.These strategies are part of an overall movement of 'turning inwards', characterized by a renewed focus on the community, the territory, and the local dimension. This movement has gone with a widespread reflection on the way of life perceived as the most suitable for the communities of Oaxaca. “Care” as a concept represents the thread of the thesis, which concludes by introducing some questions for possible future developments: which transformations occurred during the pandemic will also affect beyond Covid-19? During and beyond a pandemic, how can anthropology be part of the cure?
(RE)IMPARARE DALLA PANDEMIA: HACER COMUNIDAD DI FRONTE AL COVID-19 (OAXACA, MESSICO) / E. Fusar Poli ; tutor: S. Allovio ; coordinatore: A. Pinotti. Università degli Studi di Milano, 2023 Jul 17. 35. ciclo, Anno Accademico 2022.
QUANTITATIVE STABILITY VIA THE METHOD OF MOVING PLANES: APPROXIMATE SYMMETRY FOR OVERDETERMINED AND RIGIDITY PROBLEMS
Since its introduction thanks to the work of Alexandrov, the method of moving planes has seen a widespread use in various applications of geometric analysis. Brought to the attention of the PDE community with the seminal work of Serrin, it has been a useful tool to prove a large variety of results, including symmetry results for overdetermined and rigidity problems. This thesis investigates three such problems from a quantitative viewpoint, by employing the method of moving planes and developing tools and techniques to prove symmetry and approximate symmetry results.
QUANTITATIVE STABILITY VIA THE METHOD OF MOVING PLANES: APPROXIMATE SYMMETRY FOR OVERDETERMINED AND RIGIDITY PROBLEMS / L. Pollastro ; tutor: G. Ciraolo, M. Cozzi ; coordinator: D. Bambusi. Dipartimento di Matematica Federigo Enriques, 2024 Mar 06. 36. ciclo, Anno Accademico 2022/2023.
BIOBASED APPROACHES AT MODULATING THE INTERACTION BETWEEN LEGUMES BIOPOLYMERS AND BIOACTIVES: A PERSPECTIVE FOR THE PRODUCTION OF BAKED GOODS
Nowadays, with the increasing consumer demand for healthy foods, legumes, pseudocereals like buckwheat, and cereals like sorghum, have gained popularity as a potential component of functional foods. These crops are characterized by well-balanced nutritional profile, made by proteins, carbohydrates, resistant starch, and bioactive compounds, and offer considerable health benefits. Recent studies on buckwheat employing in vitro and ex vivo models have investigated the cellular and molecular mechanisms through which it affects human health, highlighting its capacity to reduce cytokine secretion and oxidative stress mainly related to polyphenolic and protein fractions. However, these crops are characterized by high levels of anti-nutritional factors like phytates and protease inhibitors, along with oligosaccharides found specifically in legumes, responsible for reducing the bioavailability of nutrients. To address this issue, the sprouting process on grains and legumes has emerged as a leading area of research due to its positive effect on nutritional composition. This process, applied to buckwheat (Fagopyrum esculentum), sorghum (Sorghum bicolor) and cowpea beans (Vigna unguiculata), has proven to have an effect on nutritional profiles by increasing antioxidant capacity and reducing anti-nutritional factors. Specifically, in buckwheat, sprouting for 48 and 72 hours increased the content of free phenolic compounds and peptide while decreasing significantly the level of anti-nutrients. Sorghum exhibited the same benefits with sprouting increased hydrolysis of proteins and starch and the antioxidant profiles, suggesting it could serve as a functional option in various food products, particularly in low-income regions. Cowpea also shown to benefit significantly from sprouting for 48-72 hours, which improved protein digestibility and nutrient availability while effectively lowering anti-nutritional compounds. The results of all the studies on germination across legumes and grains consistently found 72 hours to be the optimal sprouting time. Based on this, wheat bread enriched with 25% of 72h sprouted cowpea flour was prepared. Substantial nutritional improvements were observed, offering higher content of proteins and resistant starch, as well as higher antioxidant capacity for both free and bound phenolic compounds, and lower content of anti-nutritional factors. This not only improved the quality of the bread but also promoted greater protein bioaccessibility during digestion, underscoring the potential of sprouted legumes in fortifying baked goods, expanding their application in the food industry.Collectively, all these studies illustrate the impact of sprouting on various grains and legumes, improving their nutritional characteristics and viability as ingredients in functional foods. By mitigating anti-nutritional factors and improving the availability of beneficial compounds, sprouting emerges as a crucial processing technique that could emphasize how these staple crops are utilized, ultimately contributing to nutritional security and improved health outcomes worldwide. Although further research is needed, the current findings point to a promising future for sprouted flours as key elements of healthier diets, with substantial economic and nutritional advantages, especially for communities dependent on staple crops.
BIOBASED APPROACHES AT MODULATING THE INTERACTION BETWEEN LEGUMES BIOPOLYMERS AND BIOACTIVES: A PERSPECTIVE FOR THE PRODUCTION OF BAKED GOODS / S.m. Borgonovi ; tutor: M. Di Nunzio ; co-tutor: S. Iametti ; coordinator: D. Mora. Dipartimento di Scienze per gli Alimenti, la Nutrizione e l'Ambiente, 2024 Dec 20. 37. ciclo, Anno Accademico 2023/2024.
LEGAL RECOGNITION OF POLYAMOROUS FAMILIES: A CLAIM FOR DESIRABILITY
The aim of this work is twofold. Firstly, to investigate the feasibility of the legal recognition of polyamorous families from a socio-legal perspective. To achieve this purpose, in the first part of the work I outline an empirical and legal discussion of the phenomena of new families. This premise helps me deal with the issue of the feasibility of polyamorous families’ legal recognition in the long and short run. Furthermore, I point out the relevance of polyamorous relationships as a case study, by assessing their statistical prevalence, and visibility, problematising possible definitions and enumerating their main features and values. The socio-legal context of increasing visibility and social acceptance of new families, including polyamorous ones, grounds my argument in favour of the feasibility of the legal recognition of polyamory, at least in the long run. Moreover, the empirical framework further supports my normative argument in favour of the desirability of the legal recognition of polyamory.The second aim of this work is to argue in favour of the desirability of the legal recognition of polyamorous families on an equal basis, with a political-philosophical approach. Accordingly, the second part of the dissertation addresses normative questions about the legitimacy and the desirability of liberal states’ intervention in recognizing families. I argue that, both from the perspective of political and perfectionist liberalism, the law can legitimately recognize families as relationships of care. Furthermore, I foreground that states’ intervention in the matter of family is desirable as well. However, in order to be legitimate, family law should recognize all different kinds of families, including polyamorous ones. To conclude, I advocate the desirability of the legal recognition of polyamorous families both from a neutralist and a perfectionist perspective, and I suggest a necessity to look for new legal institutions, more suitable than marriage in regulating all families on an equal basis. Adopting both a socio-legal and a political-philosophical perspective, I aim to offer a systematic review of the topic of the legal recognition of polyamorous families, which has been addressed very scarcely and fragmentarily so far.
LEGAL RECOGNITION OF POLYAMOROUS FAMILIES: A CLAIM FOR DESIRABILITY / F. Miccoli ; tutor: N. Riva ; coordinatore: M. Jessoula. Dipartimento di Scienze Sociali e Politiche, 2022. 34. ciclo, Anno Accademico 2021.
RECONSTRUCTING PHOENICIAN POTTERY PRODUCTION IN SARDINIA: A MULTI ANALYTICAL CHARACTERIZATION OF SAMPLES FROM THE SITE OF PANI LORIGA (8-7TH CENTURY BC) AND MONTE SIRAI (6-5TH CENTURY BC)
This study contains the results of a multi-analytical archaeometric characterization of 53 pottery samples from two archaeological sites, Monte Sirai and Pani Loriga. Both sites are Phoenician-Punic colonies (7th-8th century BCE for Pani Loriga and 5th-6th century BCE for Monte Sirai) located in southwestern Sardinia, Italy. The analyses aimed to unveil the archaeological context, with provenance, raw materials and production processes.Seven amphora samples from Monte Sirai were first analysed as comparative materials for the largest group of samples from Pani Loriga, including amphoras, jugs, cups, plates, and pots, representative of the various shapes and types found at the site.Investigations were firstly focused on microstructure, matrix composition, and inclusion types, as well as to identify newly formed phases resulting from firing. Based on these analyses, five fabric groups were identified. Two of these groups, encompassing most of the samples, are characterized by coarse inclusions, suggesting minimal processing or purification of raw materials during the ceramic body preparation. In contrast, the other three groups display purified clay, with minimal inclusions. Optical petrography, combined with X-ray diffraction analysis, provided insights into the mineralogical associations, firing temperatures, and rock inclusions. In the most populated groups, the presence of minerals such as quartz, plagioclases, feldspars, and hydro-silicates, compatible with igneous rocks from pyroclastic and epiclastic deposits (including dacites and rhyolites), suggests the local production of these samples. Conversely, a high calcite content, along with microfossils observed in thin sections, indicates that some samples were likely imported. These imported specimens exhibit distinct features, suggesting different manufacturing techniques and, possibly, a higher level of production quality, if compared to the local ceramics. X-ray diffraction analyses further identified three distinct mineralogical groups based on their assemblages. The identification of minerals such as hydro-silicates, (primary) calcite, gehlenite, and diopside reflect different firing conditions and allow the estimation of firing temperature ranges. Such temperature ranges are further supported by archaeomagnetic analyses, which provided additional insights into the thermal history of the pottery confirming the firing process reconstructions based on XRD data (and the mineral stability fields). This first characterization allowed to select some samples to perform micromorphological analyses by scanning electron microscopy (with SEM-EDS), in order to describe matrix structure, slips and engobes, inclusion types, and degree of vitrification. SEM-EDS investigation was also combined with a portable X-ray fluorescence analysis to obtain the chemical composition of the ceramic samples. Even this approach provided experimental evidence to distinguish locally produced ceramics from potential imported ones.
RECONSTRUCTING PHOENICIAN POTTERY PRODUCTION IN SARDINIA: A MULTI ANALYTICAL CHARACTERIZATION OF SAMPLES FROM THE SITE OF PANI LORIGA (8-7TH CENTURY BC) AND MONTE SIRAI (6-5TH CENTURY BC) / G. Morabito ; advisor: G.D. Gatta ; co-advisor: N. Marinoni ; external co-advisor: C. Colombo, E. Possenti. Dipartimento di Scienze della Terra Ardito Desio, 2025 Apr 03. 37. ciclo, Anno Accademico 2024.
PATHOLOGY OF ROUND CELL TUMORS AND SARCOMAS: A DIAGNOSTIC AND CLINICALLY-ORIENTED APPROACH, WITH A FOCUS ON BASIC INVESTIGATION
Neoplasms represent a constantly increasing threat for companion animals, requiring fast and reliable diagnostic techniques. In this context, cytology is a diagnostic tool widely applied to preliminarly investigate the nature of lesions observed in daily veterinary practice. Nevertheless, considered the limitations of cytology as well as the key role that a cytological diagnosis might cover for the clinical approach to each patient, estimation of the reliability of this technique represents a fundamental step. With this in mind, diagnostic accuracy studies can provide a proof of reliability of cytological results, when the latter are compared to the histopathological gold standard. With these premises, the main aim of the current Thesis was to evaluate the diagnostic accuracy of cytology applied to different round cell tumors and sarcomas currently representing a serious threat for the canine and feline species. Chapter 1 describes a study investigating the diagnostic accuracy of cytology in the evaluation of canine splenic neoplasm. To the best of our knowledge, our work was the first study conjunctively reporting overall accuracy, sensitivity, specificity, positive predictive value, and negative predictive value of cytology in the diagnosis of these lesions. Diagnostic accuracy indexes identified limitations of negative cytological results in excluding a dog to be truly free from neoplasia; however, high specificity and positive predictive value still highlighted cytology as a valuable tool in the diagnostic approach to splenic neoplasms. In Chapter 2 is described a study investigating interobserver agreement and diagnostic accuracy of cytology in the immunophenotype prediction of feline nodal lymphoma. Our results revealed a low inter-observer agreement and a low diagnostic accuracy in immunophenotype prediction, thus highlighting potential marked differences among laboratories and even among different cytologists within the same laboratory, and consequently the mandatory need of histopathology and immunohistochemistry for a correct interpretation of feline nodal lymphoma immunophenotype. Chapter 3 describes two studies focusing on the application of cytology in the evaluation of nodal metastasis of canine mast cell tumor (MCT). The preliminary investigations described in Section 1 focused on the quantification of mast cells in cytological nodal samples obtained from both non oncological dogs and MCT-bearing dogs, revealing that mast cell (MC) quantification in lymph node (LN) cytological samples obtained from the latter might be useful to determine the nodal metastatic status. Our findings further suggested that neither the sampling technique applied to collect cytological samples nor the quantification method applied to estimate the number of MCs, influence the number of nodal MCs observed. Section 2 describes a study investigating interobserver agreement and diagnostic accuracy of cytology in the evaluation of the metastatic status of lymph nodes obtained from MCT-bearing dogs. Specifically, the study investigated the diagnostic performance of the cytological interpretative system currently available in literature as well as that of 2 amendments of the latter (AM1 and AM1.2 system). Our results revealed that the AM1.2 system, which include MC quantification besides the cytological criteria reported in the previously published cytological interpretative system, could represent a valid alternative to the latter, being characterized by an almost overlapping interrater agreement, and sensibly higher accuracy and sensitivity without substantial changes of the other diagnostic accuracy indexes. Besides studies focusing on the diagnostic accuracy of cytology, the Addendum briefly describes the results of 2 research projects investigating viral oncolysis in a cell culture model of canine histiocytic sarcoma, performed by the Ph.D. candidate during his externship at the University of Veterinary Medicine of Hannover (TiHo Hannove
PATHOLOGY OF ROUND CELL TUMORS AND SARCOMAS: A DIAGNOSTIC AND CLINICALLY-ORIENTED APPROACH, WITH A FOCUS ON BASIC INVESTIGATION / M. Gambini ; tutor: C. Giudice ; phd course coordinator: V. Grieco. Dipartimento di Medicina Veterinaria, 2021 Mar 30. 33. ciclo, Anno Accademico 2020.
"BACTERIAL PLAYERS IN CROP SYSTEMS: BENEFITS AND RISKS IN THE "" ONE HEALTH"" PERSPECTIVES."
In recent years, agri-food ecosystems have recognized as crucial environment in the One Health approach, which deals with the intersections between human, animal and environmental health, recognising them as a unique interconnected system. The research conducted in the frame of this PhD thesis dealt with the investigation on the effect of bacteria entrance in agri-food ecosystems. In compliance with the ‘One Health’ concept, both the benefits and the risks given by the introduction, voluntary or accidental, of bacterial players in environmental habitats related to food production were considered.An example of deliberate release of microbes in agricultural systems is the use of plant beneficial strains (i.e., biofertilizer and biostimulants). Experiments were designed to better clarify the interactions occurring between bacteria inoculants and the recipient plant microbiome and, additionally, the research was focused on the design of novel delivery system for the administration of plant probiotics. The knowledge improvement regarding these two aspects has the potential to promote the transition from the lab to the field of microbial based solution for sustainable agriculture, such as the replacement of chemical fertilizers or the use of strains to improve plant tolerance to biotic and abiotic stresses. We characterized for Plant Growth Promotion (PGP) activities a collection of endophytic strains established from grapevine and lettuce plants collected in the field. Among the most promising strains, Rhizobium sp. GR12 and Kosakonia sp. VR04 were tagged with genes coding for fluorescent proteins and used to inoculate micropropagated grapevine cuttings. Plantlets were grown in vitro under optimal growth conditions or on a diluted medium to mimic nutritional deficit. The measurement of plant biomass showed that Rhizobium sp. GR12 displayed PGP activity, when the plantlest were grown under nutritional deficit. The colonization of the plant tissues was confirmed for both Rhizobium sp. GR12 and Kosakonia sp. VR04 through qPCR amplification of the GFP and m-Scarlet protein, respectively. However, though both the strains were able to colonize root and lead endosphere of the treated plantlets, metataxonomic analyses revealed that the composition of the endophytic community was differently modulated by Rhizobium sp. GR12 and Kosakonia sp. VR04. The former had a minor impact on the endophytic bacterial community while Kosakonia sp. VR04 caused a major change in community composition, suggesting an opportunistic colonization pattern. The results of this study corroborate the importance of preserving the native endophytic community structure and functions during plant microbiome engineering. Moreover, for the first time, the structure of bacterial endophytic community of micropropagated grapevine cuttings was described, showing that it clearly differed from those generally associated to this plant species in the field.Rhizobium sp. GR12 strain was included in a panel of plant-associated bacterial strains isolated from different host species and selected based on their PGP properties, abiotic stress tolerance and polysaccharides degrading activity. These strains were used to test the possibility to provide beneficial bacterial inoculant through the functionalization of a bio-based polymer, derived from food industry waste and designed with the final aim to obtain biodegradable nursery pots. Cell viability within the biopolymer was evaluated by embedding the strains into the liquid material and by isolating them after its solidification, drying and storage. The results showed that the approach was suitable only for the tested spore-forming Bacillus strains, which were able to cope the desiccation step during the production of the biopolymeric film. Thus, to improve the survival of non-spore-forming strains, we changed strategy and tested food waste derived materials, and other different mol
"BACTERIAL PLAYERS IN CROP SYSTEMS: BENEFITS AND RISKS IN THE "" ONE HEALTH"" PERSPECTIVES / G. Patania ; tutor: F. Mapelli, S. Borin; coordinatore: D. Mora. Dipartimento di Scienze per gli Alimenti, la Nutrizione e l'Ambiente, 2024 Dec 18. 37. ciclo, Anno Accademico 2023/2024."
INHERENT CHIRALITY AT WORK FOR APPLICATIONS IN CHIRAL ELECTROCHEMISTRY, CHIROPTICS AND SPINTRONICS.
This three-year thesis project is aimed to deeply investigate the potentialities of the inherently chiral strategy in the chiral voltammetry field, as well as in circular dichroism and magnetoelectrochemistry ones. Voltammetric experiments must take place at chiral electrochemical interphases with the implementation of (inherently) chiral selectors, according to two strategies: i) exploitation of electrode surfaces modified with electrodeposited heterocycle-based inherently chiral oligomer films (with the stereogenic element responsible for chirality coinciding with the functional group responsible for the material specific properties); ii) use of enantiopure chiral media with high intrinsic order.Concerning the modification of electrodes with inherently chiral oligomer films, several new inherently chiral monomers were studied with different techniques, in order to amplify the range of monomer structures studied and the combination of chiral selector/probe investigated (in fact, many new chiral probes, also of pharmaceutical interest were used in enantioselection tests, with different nature, chemical structures, bulkiness and reactivity). Enantioselection tests were discussed as a function of the experimental conditions; moreover, discussion of experiments with a systematic series of planar stereogenicity ferrocenes provided key clues concerning the probe-selector interactions in the process, while experiments on a systematic new family of inherently chiral monomers with different substituents on the N atoms in the biindole core enabled to study the effect of H bonds in the enantioselection process. Finally, the inherently chiral oligomer films were discussed in advanced application for magnetoelectrochemistry experiments, employing an innovative experimental setup and resulting in impressive spin-related effects. Also concerning the chiral media approach, several new chiral selectors were presented, employing them as low concentration chiral additives in achiral commercial ionic liquids or as bulk media, testing their performances in enantioselection experiments. The performances of bio-based and synthetic chiral selectors were compared, studying different artificial inherently chiral media. The application of another kind of advanced chiral media were exploited, studying the enantioselection ability and properties of chiral Deep Eutectic Solvents (DESs) of natural origin. The DESs employed in this research project are composed of a mixture of at least two components, a hydrogen bond acceptor (HBA) and a hydrogen bond donor (HBD), which are able to self-associate to form a new eutectic phase characterized by a melting point (< 100°C) lower than that of each individual component and significantly lower than the predicted one for an ideal case.
INHERENT CHIRALITY AT WORK FOR APPLICATIONS IN CHIRAL ELECTROCHEMISTRY, CHIROPTICS AND SPINTRONICS / S. Grecchi ; tutor: P. R. Mussini ; supervisori: T. Benincori, S. Arnaboldi. Dipartimento di Chimica, 2022 Mar 25. 34. ciclo, Anno Accademico 2021. [10.13130/grecchi-sara_phd2022-03-25].
MODERN ANIMAL NUTRITION IN THE ERA OF SUSTAINABLE DEVELOPMENT GOALS
The livestock sector faces several challenges for 2050: the growth of the human population and the consequent greater demand for products of animal origin, the reduction of the environmental impact and the use of natural resources, as well as the reduction of feed/food competition. In this scenario, animal husbandry offers the environment ecosystem services, and the supply of protein and essential micronutrients for populations, especially in developing countries. The sector of feed production and livestock breeding is constantly evolving, numerous technologies are being developed in the field of animal nutrition, for example, nutrigenomics and omics techniques to evaluate feed ingredients for animal diets. In particular, metabolomics, i.e. the study of all the metabolites present in the bio-fluids of an organism, is under development since it offers the advantage of carrying out matrice samplings in a non-invasive way and offering a complete analysis of the animal metabolic profile. As far as the challenges of the feed sector are concerned, the keyword remains feed safety, which refers mainly to mycotoxin contamination. Mycotoxins are ubiquitous, and it is impossible to prevent their development, but it is essential to apply preventive and risk management protocols throughout the entire feed production chain. In a scenario of climatic changes, it is essential to study their effects on mycotoxin contamination in crops. Climate changes are to be considered another challenge for the livestock sector which is often labeled as responsible for greenhouse gas emissions, including methane, nitrous oxide, and carbon dioxide. It is essential to accurately assess emissions from the livestock sector and feed production. As efforts need to be made to mitigate environmental emissions and enhance food industries' by-products ensuring the transition to a circular economy, one proposal is the use of alternative sources foranimal nutrition, such as former foodstuffs and by-products and co-products derived from food industries. Evaluating the quality and nutritional characteristics of new ingredients is one of the current challenges in the feed industry. The application of new technologies in the field of animal nutrition research is one of the future objectives, with the aim of improving the quality of animal origin products, ensuring animal health and welfare as well as the ongoing development of the livestock sector from a precision nutrition perspective to face modern challenges.
MODERN ANIMAL NUTRITION IN THE ERA OF SUSTAINABLE DEVELOPMENT GOALS / F. Fumagalli ; tutor: F. Cheli, L. Pinotti ; coordinatore: L. Pinotti. Università degli Studi di Milano, 2023 Apr 13. 35. ciclo, Anno Accademico 2022.
CRIMINALITA' ORGANIZZATA CALABRESE TRA PERIODO LIBERALE E FASCISMO NELLE PROVINCE DI REGGIO CALABRIA E CATANZARO.
Abstract This research investigates the history of organized crime in the provinces of Catanzaro and Reggio Calabria, with particular attention to the district of Monteleone (today part of the province of Vibo Valentia), between the late nineteenth century and the Fascist period. The aim is to reconstruct, through the analysis of judicial sources, archival documents, and contemporary press materials, the dynamics of formation, diffusion, and repression of criminal organizations active in this area.Long considered a “minor” mafia, Calabrian organized crime has only recently attracted the attention of historians. This study fits into a line of research that seeks to fill the gaps in the historiography of the ’ndrangheta in peripheral areas compared to the better-studied Reggio Calabria context. In particular, it examines the evolution of the so-called picciotteria and its transformations over time, highlighting a ritual and symbolic continuity with the modern forms of the ’ndrangheta.The investigation begins with an analysis of judicial proceedings held between the late nineteenth and early twentieth centuries, which reveal the presence in the Monteleone district of true camorrist “sects,” often interconnected with the criminal networks of the Gioia Tauro plain, especially with Palmi. Even in late-nineteenth-century documentation, we find ritual elements, specific terminology, hierarchical structures, and operational practices that anticipate the organizational model later attributed to the ’ndrangheta. Some trials reveal a structure divided into ranks (picciotti and camorristi), initiation rites involving symbolic use of blood and honor, and a territorial network encompassing various municipalities in the region.One of the most significant aspects emerging from the research concerns the role of spaces of forced coexistence—prisons, barracks, public worksites—in the dissemination of criminal models. Judicial records and testimonies show that the military environment, during and after the First World War, served as a privileged space for the transmission of mafia knowledge, as evidenced by the case of the “Società di Onore e Sangue di Pachì” (“Society of Honor and Blood of Pachì”). Special attention is also given to the role of women within ritual and operational contexts—an often overlooked yet recurrent element.The postwar period and the rise of Fascism marked a phase of reorganization for organized crime, but also of transformation in the political and judicial landscape. Fascist repression of Calabrian organized crime, although less systematic than in Sicily, was nonetheless documented in Monteleone and surrounding areas. However, the limited number of convictions for criminal association and the involvement of influential figures in the defense of the accused (such as lawyer and politician Ignazio La Russa) raise questions about the real effectiveness of Fascist repression and the possible political protection enjoyed by local criminal groups.The thesis also addresses the internal structure of the ’ndrangheta during this period, engaging with the historiographical debate over the existence of hierarchical leadership bodies—such as the “Criminale di Polsi” or the “Gran Criminale”—capable of exercising coordinating or judicial authority. While some testimonies support the idea of a structured and centralized organization, other sources suggest a more horizontal network of autonomous yet culturally homogeneous local groups.Ultimately, this research interprets Calabrian organized crime as a territorially rooted social phenomenon. The picciotteria thus appears as a historically and culturally continuous criminal model, anticipating many of the defining features of the modern ’ndrangheta. The proposed interpretative hypothesis points to a shared genesis of the Italian mafias (Camorra, Cosa Nostra, and ’Ndrangheta), ori
CRIMINALITA' ORGANIZZATA CALABRESE TRA PERIODO LIBERALE E FASCISMO NELLE PROVINCE DI REGGIO CALABRIA E CATANZARO / R. Pileggi ; tutor: F. Basile ; cotutor: C. Castellano ; coordinatore: F. Basile. Dipartimento di Scienze Giuridiche Cesare Beccaria, 2025 Oct 30. 35. ciclo, Anno Accademico 2024/2025.
TECHNOLOGICAL BREAKTHROUGH TOWARDS THE USE OF A NOVEL VISUAL-INERTIAL ODOMETRY SYSTEM AS AN AID FOR THE DIGITALLY-GUIDED INTERVENTION
Recent data availability of three dimensional imaging brought new possibilities for the guided intervention. The management of complex data requires the use of new tools and new technologies. We analyzed the use of algorithms for the automatic, ai-driven segmentation to extrapolate the best systems for the use of CBCTs. The tool improves the timing and precision of the segmentation and allows to give normal values for the cephalometry. The segmented data can be analyzed through three dimensional alignment, the new techniques will be explored for data synthesis. Finally three dimensional resulting datasets can be exported and used to virtually plan an intervention. After the plan is complete they can be used to bring the clinician a new powerful system for the AR guided intervention. Clinical and research data results and outcomes will be given, bringing both preliminary as well as multidisciplinary researchers in the field of: orthodontics, endodontics and oral surgery with clinical direct performances and results.
TECHNOLOGICAL BREAKTHROUGH TOWARDS THE USE OF A NOVEL VISUAL-INERTIAL ODOMETRY SYSTEM AS AN AID FOR THE DIGITALLY-GUIDED INTERVENTION / M. Farronato ; tutor: C. Maiorana ; coordinator: M. Del Fabbro. Dipartimento di Scienze Biomediche, Chirurgiche ed Odontoiatriche, 2022 Dec 14. 35. ciclo, Anno Accademico 2022.
GENETIC IMPROVEMENT OF CAMELINA SATIVA: DEVELOPMENT OF NEW VARIETIES AND EXPLORATION OF ITS POTENTIAL AS AN INTERMEDIATE CROP
In recent years, Camelina sativa has gained considerable attention as a sustainable crop, known for its resilience under harsh conditions and low input requirements. It is being explored for its potential as a biofuel source and for the production of omega-3 rich oil. Furthermore, its role in improving soil health through crop rotation and its environmental benefits, such as reducing greenhouse gas emissions, have further increased its attractiveness. This doctoral thesis presents an overview of research on the allelopathic and agronomic benefits of Camelina, in particular its potential in integrated management. The thesis examines progress in camelina breeding through traditional methods, biotechnological approaches, evaluating its potential to improve the crop (improvements in yield, oil content and reduction of anti-nutritional compounds such as glucosinolates). He underlines the importance of genetic diversity and proposes two selection strategies: one that involves crossing spring varieties and the other that crosses spring and winter varieties. Breeding programs have shown that winter cultivation in Northern Italy led to significantly higher yields than spring cultivation. Furthermore, a synthetic population of Camelina sativa was identified as adaptable with a low glucosinolate content, making it resilient to different environments. Molecular analysis made it possible to identify genetic differences between winter and spring varieties. Finally, the promising new variety (C1244) shows early maturity, high seed weight and oil content, offering potential for use in several sectors.
GENETIC IMPROVEMENT OF CAMELINA SATIVA: DEVELOPMENT OF NEW VARIETIES AND EXPLORATION OF ITS POTENTIAL AS AN INTERMEDIATE CROP / M. Ghidoli ; tutor: S.R. Pilu ; cotutor: F. Adani ; coordinatore: S.R. Pilu ; supervisori: E. Cominelli, E. Nielsen. Dipartimento di Scienze Agrarie e Ambientali - Produzione, Territorio, Agroenergia, 2024 Dec 16. 37. ciclo, Anno Accademico 2023/2024.
MULTISYSTEM INFLAMMATORY SYNDROME IN CHILDREN:LONGITUDINAL BODY COMPOSITION ASSESSMENT, INFLAMMATORY MARKERS, AND FATTY ACIDS PROFILE
The Sars-Cov 2 pandemic mainly affected the adult population. However, a small portion of children were affected by Multisystem Inflammatory Syndrome in Children (MIS-C). The severity of the syndrome required patients to be hospitalised and, in some cases, admitted to pediatric intensive care units. A high risk of malnutrition was detected in children hospitalised for long periods. In addition, omega-3 fatty acids (ω-3 FA) were used in conjunction with drug therapy in adults hospitalised for Covid-19 to improve the inflammatory response. The aim of this work was to assess the risk of malnutrition in the acute phase and body composition over time in a cohort of children diagnosed with MIS-C. For all the recruited children, inflammatory markers, metabolic parameters, and fatty acid levels were assessed both in the acute phase and six months after discharge, alongside anthropometric assessment. In half of the patients recruited, three months of docosahexaenoic acid (DHA) supplementation was also recommended. We observed a change in body composition in the acute phase, with a reduction in fat-free mass. In contrast, in the long term we detected a gain in fat mass, probably due to the limited spontaneous and structured physical activity on the part of the children. The fatty acid levels detected in the acute phase were lower than in the healthy population, while the glucose profile was altered. At three and six months after discharge, the ω-3 FA values were higher than the acute phase, particularly in the group that received supplementation. The correlation between ω-3 FA levels and inflammatory indices at six months after discharge in the supplemented group, showed a positive correlation between ω-3 eicosapentaenoic acid (EPA) and erythrocyte sedimentation rate (ESR)and d-dimer, as well as a positive correlation between DHA and ferritin. Overall, the MIS-C children had a favourable disease course. However, nutritional status and loss of body weight should always be monitored in this type of disease during hospitalisation. In the long term, adherence to a balanced diet should be advised along with the resumption of spontaneous and planned physical activity. At the same time, ω-3 FA supplementation may facilitate optimal recovery. As a future perspective, the effectiveness of ω-3 supplementation in the acute phase and later, in seriously ill paediatric patients should be explored.
MULTISYSTEM INFLAMMATORY SYNDROME IN CHILDREN:LONGITUDINAL BODY COMPOSITION ASSESSMENT, INFLAMMATORY MARKERS, AND FATTY ACIDS PROFILE / E. Di Profio ; tutor: E. Verduci ; coordinator: L. Pinotti. Dipartimento di Scienze della Salute, 2023. 36. ciclo, Anno Accademico 2023.
GENOMIC APPROACHES AND PHENOTYPIC ANALYSES FOR IMPROVING THE SELECTION OF FRUIT QUALITY TRAITS IN PEACH [PRUNUS PERSICA L. (BATSCH.)] AND APRICOT (PRUNUS ARMENIACA L.)
Peach [Prunus persica L. (Batsch.)] and apricot (Prunus armeniaca L.) are two Prunus species developing delicious fruits and they are mostly grown in temperate areas of the world. Both species have been cultivated since ancient times, being greatly appreciated for their unique taste and benefits to human health. Peaches and apricots can be consumed either fresh or processed through canning or drying, depending on the preference of a specific region and the use-purpose. So far, many efforts have been done in enhancing disease and pest resistance, in increasing the environmental adaptability to sub-tropical areas, in extending the harvest season or in improving fruits appearance. However, consumers frequently complain about the lack of taste in peach and apricot fruits sold on markets, encouraging the ongoing breeding projects to include organoleptic properties among their objectives. Among all the fruit quality-related traits, acidity plays a pivotal role affecting both consumers’ and market acceptance. To match with market trends, an extensive characterization of peach and apricot fruits attributes under the highlighted necessity of renewing the varieties cultivated worldwide is required. In this thesis, a peach and apricot collection of 201 and 164 accessions, respectively, was screened for many important fruit organoleptic attributes, with a specific focus on acidity and organic acids content. Fruits acidity was titrated and ten organic acids (cis-aconitate, citrate, fumarate, galacturonate, malate, oxalate, quinate, shikimate, succinate and tartrate) separation was accomplished by HPLC technique coupled to UHPLC-HRMS validation. Analyses were performed on peach pulp and on apricot pulp and peel. The final aim was in-depth dissecting the peach and apricot panel for these organoleptic parameters to exploit the existing variability within their germplasm. Malate and citrate were the most abundant organic acids in both species, with pattern more genotype than year dependent. Results suggested that seasonality effects on fruits acidity and almost all the considered-organic acids were very low. Among the other organic acids, quinate and succinate reached large concentrations in peach and apricot, respectively, while tartrate was interestingly present more in apricot peel than pulp. The availability of a reference genome in peach has allowed to further characterize peach fruit quality traits. Over the past, peach fruits acidity has been partially elucidated at genetics level, identifying one major locus and making it a breeding target. Although the great advances, the selection accuracy and the long-time required for releasing new varieties on markets still hamper peach breeding progress. To in-depth unravel acidity trait and to speed up the selection of newly developed individuals in peach, a total of 1,190 accessions were genotyped for performing genomics analyses. Two different studies were carried out in this thesis: genomic selection (GS) and genome wide association studies (GWAS). Results confirmed that GS seems feasible in peach not only for acidity but also for organic acids content, in particular for patterns of malate and citrate. GWAS confirmed the presence of one major locus acting as dominant in peach fruits but revealed other significant associations on chromosomes 1, 7 and 8. In summary, this thesis includes the first efforts in an in-depth and at multilevel dissection of acidity in peach and apricot, applying different approaches on a large panel of individuals. This thesis attempts to provide a complete overview with results that may be useful for future researches, studies and successful breeding programmes.
GENOMIC APPROACHES AND PHENOTYPIC ANALYSES FOR IMPROVING THE SELECTION OF FRUIT QUALITY TRAITS IN PEACH [PRUNUS PERSICA L. (BATSCH.)] AND APRICOT (PRUNUS ARMENIACA L.) / I. Baccichet ; tutor: A. Spinardi ; co-tutor: M. Cirilli ; coordinatore del corso di dottorato: D. Bassi. Dipartimento di Scienze Agrarie e Ambientali - Produzione, Territorio, Agroenergia, 2021 Mar 09. 33. ciclo, Anno Accademico 2020.
ASSESSING PREDICTIVE MODELING APPROACHES FOR OPTIMIZING THERAPY IN ADVANCED LARYNGEAL AND HYPOPHARYNGEAL CANCERS
Background: Laryngeal (LC) and hypopharyngeal (HPC) cancers are often diagnosed at an advanced stage (III or IV) due to the absence of early symptoms and biomarkers. First-line therapies include both surgical and non-surgical approaches. Since surgical removal, comprising total and partial laryngectomy, is highly invasive, organ-preservation strategies are frequently preferable as they help maintain organ functionality. Induction chemotherapy (ICT) is one such preservation approach; however, it is not suitable for all patients and may contribute to drug resistance in some cases. Identifying a molecular signature of ICT response in both responsive and non-responsive patients could enable prediction of ICT outcomes in patients with advanced disease, providing valuable insights for therapy decisions. Similarly, a predictive model for organ functionality could help identify patients who may benefit from alternative approaches. Moreover, exploring new combined therapies for non-responsive patients could improve survival outcomes.Methods: We conducted a retrospective analysis on a cohort of FFPE samples from patients with advanced LC and HPC treated solely with ICT across various European hospitals. Laser capture microdissection was used collecting samples enriched in tumoral cells, followed by RNA-extraction, library preparation, and RNA-sequencing. Leveraging machine learning algorithms, we generated three predictive models: (i) a combined model integrating ICT response and LED, (ii) a LED-only model, and (iii) an ICT-response model. Models were trained with iterative validation across 100 random train-test splits to ensure robustness and generated using transcriptomic, clinical, and integrated data. In addition, in vitro cisplatin-resistant cell models were established to investigate resistance-associated genes, which were subsequently integrated into predictive models. Finally, immune-related gene expression was assessed to explore the potential integration of immunotherapy.Results: Survival analyses demonstrated that ICT response strongly correlated with clinical outcomes (OS and PFS), with resistant patients showing significantly poorer prognosis. Similarly, loss of laryngo-esophageal functionality (LED) was also associated with reduced survival and quality of life, underscoring the need for functional prediction alongside therapeutic response. The combined model integrating ICT response and LED achieved high predictive accuracy, with particularly low rate of misclassification when transcriptomic data were included. The LED model demonstrated that molecular data, rather than clinical variables, drive the prediction of organ functionality, reaching robust accuracy without improvement from clinical data integration. Conversely, the ICT-response model significantly benefited from the inclusion of both clinical and molecular features, achieving AUC values above 80%, and further improved by incorporating cisplatin-resistance genes, such as OLFM4 and MGMT, identified in vitro. Importantly, immune profiling revealed that most tumor exhibited an immune-“COLD” phenotype, although a subset of partial responders with preserved organ functionality displayed a “HOT” phenotype, suggesting that ICT combined with immune checkpoint inhibitors could represent a promising strategy for selected patients.Conclusions: This study demonstrates the feasibility of integrating RNA-sequencing data and machine learning to predict ICT response and post-treatment functionality in LC and HPC. The combination of molecular and clinical data allows a more accurate patient stratification, supporting the development of personalized therapeutic strategies and the implementation of predictive biomarkers in clinical practice.
ASSESSING PREDICTIVE MODELING APPROACHES FOR OPTIMIZING THERAPY IN ADVANCED LARYNGEAL AND HYPOPHARYNGEAL CANCERS / M. Compagnoni ; tutor: S. Chiocca ; co-tutor: S. Minucci ; phd coordinator: D. Pasini. Dipartimento di Oncologia ed Emato-Oncologia, 2025 Dec 17. 37. ciclo, Anno Accademico 2024/2025.
GIUDICI, ZANZOTTO, RABONI. AUTONOMIE ED ETERONOMIE DELLA PROSA
"This dissertation investigates the non-fiction production of three Italian poets of the second half of the twentieth century: Giovanni Giudici, Andrea Zanzotto, and Giovanni Raboni. Adopting a transdisciplinary approach, it shows how these writings, often considered a ""secondo mestiere”, according to Montale's definition, do not play a merely ancillary or vicarious role in the interpretation of the poetical work of their authors, but they prove crucial for the understanding of their intellectual figures. Following an Introduction that provides essential contextual details and describes the aims, scope, and method of the dissertation, Part One looks at the post-1991 non-fiction writings of these three authors, linking them to the end of a whole Weltanschauung. What emerges is a sense of the awareness that literary criticism and essay writing in general developed of political dynamics in the contemporary world. Part Two identifies and scrutinises this textual corpus from its beginnings, datable between the Liberation and the Economic Boom, considering Giudici, Zanzotto, and Raboni’s non-fiction and literary criticism books as macro-texts and a means of verification of the season they were written in. The Third and final Part analyses some newly-recovered unpublished, scattered, or unsigned essays and highlights their significance by framing them as part of the intellectual reflection of their authors."
GIUDICI, ZANZOTTO, RABONI. AUTONOMIE ED ETERONOMIE DELLA PROSA / M. Cappello ; tutor: L. Neri ; coordinatore: F. Slavazzi. Dipartimento di Studi Letterari, Filologici e Linguistici, 2022 Jul 01. 34. ciclo, Anno Accademico 2021.
FREEDOM AS NON-DOMINATION AND EQUALITY: A NEO-REPUBLICAN THEORY OF DISTRIBUTIVE JUSTICE
My main claim in this thesis is that, if we want the fundamental ideal of neo-republicanism, notably freedom as non-domination, to be ensured, we need excessive individual wealth to be limited. This claim stems from a twofold premise. First, freedom as non-domination is constituted by political equality. To be free from domination, people should be protected from anyone else’s arbitrary capacity to interfere with their choices, that is, they should be granted an equal status to one another by the state; nonetheless, to avoid the state’s interferences to be arbitrary themselves people should also have equal opportunities to influence the public decision-making process, that is, they should be granted political equality (Pettit, 1997; 2012; Pansardi, 2013; 2015; Gädeke, 2020; 2020a). Second, though, such a political equality is jeopardized by the presence of very rich people within a democracy: very rich people indeed enjoy disproportionate opportunities to influence politics because of their wealth (Gilens & Page, 2014; Cagé, 2020). Moreover, neo-republicans have either overlooked this problem (e.g., Pettit, 2012), or they have addressed it from a procedural perspective, i.e., by envisaging institutional measures to prevent the wealthy from playing such a significant role in politics (McCormick, 2011; 2019). By contrast, I show that the presence of economic elites within a democracy represents one of the major threats for freedom as non-domination: if people do not enjoy equal opportunities to influence the lawmaking process, they shall comply with arbitrary laws, thus they will be dominated. Furthermore, I argue that procedural solutions are not sufficient to solve this problem. When people possess large amounts of wealth more than their fellows, indeed, they can easily translate it into political power through a great variety of direct and indirect mechanisms that elude formal separations between economic and political spheres (Knight & Johnson, 1997; Christiano, 2010; 2012; Robeyns, 2017). Therefore, if one wants freedom as non-domination to be ensured, one needs not only to protect the democratic ideal of political equality from the super-rich’s unfair influence, but also to investigate material, rather than formal, solutions for this purpose. This is the reason why my dissertation aims at developing a theory of distributive justice for neo-republicanism. In this respect, I argue that a limitarian principle should be advocated, namely that principle according to which no-one should have more than a certain threshold of wealth (Robeyns, 2017; 2019; 2022). My view of this threshold, however, differs from the classical limitarian formulation (Robeyns, 2017), which has also been retrieved in the first proposal of republican limitarianism (Dumitru, 2020). Rather than being put where people no longer need their resources for their full flourishing, a limitarian threshold should be put where the risks it aims at eliminating materialize (Harel Ben-Shahar, 2019). When it comes to freedom as non-domination, this means that what should be withdrawn are all those resources that allow the super-rich to enjoy disproportionate and boundless power in politics – or so I argue. Moreover, I believe that this could be done through a top marginal taxation rate of 100% on what people inherit beyond that threshold. This is because, on the one hand, a top marginal taxation rate of 100% seems to best represent the ceiling to individual wealth that a limitarian threshold aims at establishing (Robeyns, 2017). On the other, I argue that taxing inheritance away would tackle the root of the problem of political domination by economic elites, given that inheritance facilitates the accumulation and concentration of wealth in the hands of few people (Piketty, 2014; Halliday, 2018).My conclusion hence is that a limitarian theory of distributive justice, which could be implemented via an inher
FREEDOM AS NON-DOMINATION AND EQUALITY: A NEO-REPUBLICAN THEORY OF DISTRIBUTIVE JUSTICE / E. Icardi ; supervisore: N. RIVA ; coordinatore: M. JESSOULA. Università degli Studi di Milano, 2023 May 18. 35. ciclo, Anno Accademico 2022.
ELUCIDATING THE ROLE OF HET0016 IN AMELIORATING 20-HETE INDUCED OXIDATIVE STRESS AND LIVER INJURY IN B-THALASSEMIA: A NOVEL THERAPEUTIC TARGET
Background: Reactive oxygen species (ROS) are one of the main contributors to pathological outcomes in β-thalassemia, with their generation being a consequence of iron overload and abnormal red blood cell metabolism. We have previously shown that CYP450 mediates ROS production in the liver of a mouse model of β-thalassemia through an increase in 20-hydroxyeicosatetraenoic acid (20-HETE) activity.Aim: The aim of this study is to assess the efficacy of N-Hydroxy-N'-(4-butyl-2-methylphenyl)-formamidine (HET0016), an inhibitor of 20-HETE formation, in reducing 20-HETE-mediated oxidative stress and injury in the livers of mice affected by β-thalassemia.Methods: Hbbth3/+ mice (The Jackson Laboratory) were used as a model of non-transfusion dependent thalassemia (NTDT) and were divided into 3 groups: a control group (N=11) treated intraperitoneally (IP) with a 1:1 solution of dimethyl sulfoxide (DMSO) and phosphate-buffered saline (PBS), Hbbth3/+ mice treated with a 1:1 solution of DMSO and PBS (N=14), and Hbbth3/+ mice treated with HET0016 (N=7). HET0016 was administered in the form of a daily IP injection at a dose of 5mg/kg/day for a total duration of four weeks. Enzymatic activity of NADPH oxidases was assessed using the NADPH oxidase assay. mRNA expression levels of two isoforms of CYP4A, as well as CYP4A protein levels, were measured in the liver using Real-time Polymerase Chain Reaction (PCR) and western blot, respectively. Degree of expression of CYP4A was also evaluated in the liver by immunohistochemistry (IHC). Formalin-fixed paraffin-embedded liver tissues were also sectioned and stained with Pearl’s Prussian blue stain to detect ferric iron (Fe3+). Sirius red stain and measurement of hepatocyte growth factor (HGF) mRNA expression levels were performed to detect signs of liver injury. To assess the degree of lipid peroxidation, an IHC staining for 4-hydroxynonenal (4-HNE) in addition to a malondialdehyde (MDA) colorimetric assay was performed. IHC staining for Caspase-3 was conducted to assess apoptosis. Finally, the effects of 20-HETE inhibition on the ferroptosis pathway were examined by measuring the mRNA and protein expression levels of glutathione peroxidase 4 (GPX4) in addition to measuring total glutathione (GSH) levels. Results: In Hbbth3/+ mice, there was an increase in intracellular ROS levels and NADPH oxidase enzymatic activity. At both mRNA level and protein level, in addition to IHC staining, a significant increase was observed in the expression of CYP4A in Hbbth3/+ mice compared to control. This significantly decreased after HET0016 administration. Moreover, Hbbth3/+ mice showed an increase in the lipid peroxidation markers MDA and 4-HNE, which decreased after HET0016 administration. HET0016 also ameliorated liver injury in Hbbth3/+ mice treated with HET0016 as shown by a significant decrease in HGF mRNA expression levels and collagen fiber content. Increased tissue iron levels were detected in the liver of Hbbth3/+ mice compared to controls and this decreased with HET0016 administration. Caspase 3 levels also decreased following treatment with HET0016. Finally, HET0016 treated Hbbth3/+ mice showed a significant increase in GPX4 at both mRNA and protein level in addition to increased GSH levels, when compared to their non-treated counterparts. Conclusion: Our preclinical study is the first to assess the efficacy of HET0016 in reducing 20-HETE-mediated oxidative stress and injury in the livers of mice affected by β-thalassemia. These findings lay the groundwork for future and further investigations on the role of 20-HETE induced oxidative stress in β-thalassemia and may pave the way for a new novel therapeutic target in the field.
ELUCIDATING THE ROLE OF HET0016 IN AMELIORATING 20-HETE INDUCED OXIDATIVE STRESS AND LIVER INJURY IN B-THALASSEMIA: A NOVEL THERAPEUTIC TARGET / R. Bou Fakhredin ; tutor: I. Motta co-tutor: F. PeyvandiPhD program coordinator: C. Sforza. Dipartimento di Scienze Cliniche e di Comunità, 2025 Jan 23. 37. ciclo
PER UN'IDEA DI MERITO: UNA RICOSTRUZIONE CONCETTUALE E UNA PROPOSTA TEORICA
The research intends to contribute to the debate on desert, a controversial, problematic with unclear borders, but equally diffused and used both historically and currently. The aim of the thesis is to clarify the uses, meanings and contexts in which the attribution of merit is appropriate. The work maintains a scientific approach of analysis, supporting the usefulness as well as the necessity of the concept of desert for communities: the result is a framework for which desert is useful as a principle of justice and necessary as a mode of relation.Structured in three chapters, the work deals with the historical and conceptual reconstruction of merit, the relationship with responsibility and finally the relationship with equality. The first chapter responds to the need to investigate the nature of the concept, the reasons for its pervasiveness in the public debate, the relationship with justice, distributive and retributive. The second chapter deals with free will and luck, and then goes deep into the types of responsibility connected to the idea of merit. The concluding chapter deals with the thorniest elements of the debate, investigating which elements make the concept admissible for an egalitarian perspective, in particular that of relational egalitarianism, through the analysis of the concept of power and respect.Keeping firmly within the boundaries within which desert can act without generating and perpetuating injustice, the thesis proposes a way of investigating and deepening the concept in harmony with, not in opposition to, egalitarian values.
PER UN'IDEA DI MERITO: UNA RICOSTRUZIONE CONCETTUALE E UNA PROPOSTA TEORICA / G. Balossino ; tutor: C. Del Bò ; coordinatore: F. Poggi. Università degli Studi di Milano, 2023. 36. ciclo, Anno Accademico 2023.
'NDRANGHETA E SOCIETA' REGGINA FRA IL 1943 E IL 1970.
"This research examines the evolution of the ‘ndrangheta within Reggio Calabria’s society between 1943 and 1970, focusing on its relationships with politics, the economy, and law enforcement. Rather than viewing the ‘ndrangheta as a static entity, the study reconstructs how it was perceived, represented, and transformed alongside broader societal changes.The thesis is structured into seven chapters. The first defines the concept of mafia, tracing the historical and academic debate on the subject. The second outlines the methodological approach, discussing the challenges of studying organized crime through historical sources. The third reviews existing literature, highlighting gaps in research on the ‘ndrangheta’s development before the 1970s. The fourth examines the immediate postwar years (1943-1950), a period of widespread violence where criminal groups operated alongside political and social unrest. The fifth and sixth chapters analyze the 1950s, when the ‘ndrangheta strengthened its ties with local politics, particularly through electoral influence and control over labor markets. Special attention is given to the 1955 ""Operazione Marzano,"" a major police crackdown that briefly brought the organization into public focus. The final chapter explores the 1960s, a decade of transformation marked by economic expansion, increasing violence, and a growing state response, culminating in the 1967 Locri massacre and the 1969 Montalto summit.By integrating political, economic, and social history, this study challenges simplistic narratives that depict the ‘ndrangheta as an isolated or purely criminal phenomenon. Instead, it demonstrates how the organization evolved in response to shifting historical conditions, shaping and being shaped by the society around it."
'NDRANGHETA E SOCIETA' REGGINA FRA IL 1943 E IL 1970 / R. Clemenza ; tutor: C. Castellano ; co-tutor: F. Basile. - Milano. Università degli Studi di Milano, 2025 Apr 15. 36. ciclo, Anno Accademico 2023/2024.
ROLE OF A SPECIFIC NANOG SUPER-ENHANCER ERNA IN EPIGENETIC REMODELING AND ENHANCER-PROMOTER INTERACTION
During last years, the role of enhancer RNAs has been in part explored; however, it’s still not clear the exact role these transcripts carry out concerning gene regulation.In this work, the role of eRNAs transcribed from the region 5kb upper the TSS of Nanog gene in mESCs has been investigated. In particular, a specific eRNA transcript, here defined as antisense eRNA, resulted actively involved in the maintenance of Nanog gene in an active epigenetic state. After the antisense eRNA silencing, Nanog was turned-off and the cell underwent differentiation. This process was achieved through the involvement of the three layers of epigenetic regulation: the hypermethylation of the Nanog promoter and the SE region; the chromatin switch between activation and repression markers; the disruption of the SE-promoter loop responsible of the Nanog gene activation in stemness conditions. Thus, results suggested that the expression of the antisense eRNA has a key role in preserving the pluripotency in mESCs. Futhermore, preliminary experiments were performed on human cancer stem cells models to investigate the presence of eRNAs in a putative enhancer region 3kb upper the NANOG TSS.
ROLE OF A SPECIFIC NANOG SUPER-ENHANCER ERNA IN EPIGENETIC REMODELING AND ENHANCER-PROMOTER INTERACTION / D. Costabile ; supervisor: L. Chiariotti ; additional supervisor: M Cuomo. Dipartimento di Oncologia ed Emato-Oncologia, 2025 Feb 21. 36. ciclo
THE CONSERVATION AND PROMOTION OF THE 'CULTURAL HERITAGE OF MANKIND': WHICH OBLIGATIONS UNDER INTERNATIONAL LAW?
The research consists in an international law-based analysis which, focusing on the issue ofdetermining the obligations set up, notably, by the United Nations, towards the protection of the‘cultural heritage of mankind’ pending on the international community, approaches the enhancementof the human right to culture in an intergenerational and sustainable-led perspective.In detail, the research consists in a study of the current international framework for the worldwideprotection of cultural heritage put in place, notably, under the aegis of UNESCO, and applicable toStates Parties both in peace time and in war. In this sense, the research tries, first of all, to reconstructthe existing international norm set established, notably, by United Nations and UNESCO, for theinternational protection of the cultural heritage of peoples, thereby focusing on the analysis of theobligations pending on the international community according to such a norm set. In this view, theanalysis aims at the study of the scope and features of such existing duties pending on States Partieswith regard to the protection of the world’s cultural goods and sites according to UNESCO both inpeace time and in war, focusing, notably, on the provisions of the UNESCO 1954 Hague Conventionand of the UNESCO 1972 World Heritage Convention. In particular, the research concentrates on theinternational framework established by the global community for the worldwide protection of culturalheritage possibly endangered in times of peace, and, notably, by intentional destruction – this, evenmore, in view of the several recent episodes of ‘iconoclastic propaganda’ which have been occurring,since the first decade of 2000s, in many areas of the globe.In line with the interdisciplinary approach at the core of the Ph.D. Programme, the above-mentionedcurrent UNESCO framework is approached through the means of an integrated perspective, and,notably, through the lens of anthropological studies – and, in particular, of legal anthropology andcultural anthropology.Coming up, as the result of the analysis, with an overview on the current scope and features, as wellas of the remarkable existing discrepancies and intrinsic shortcomings, currently entailed within theUNESCO norm-set for cultural heritage protection in peacetime, the study identifies as its coreobjective the possible identification of feasible solutions to cope with such current normative aporia.In this sense, the work shifts to the study of the human rights framework for the protection of culturalrights, approached, in this context, in an intergenerational perspective. In this view, the researchfocuses on the significance of ensuring adequate international protection to cultural heritage as acomponent of the human right to culture, pursuant to art. 15 para. 1 lett. a) of the ICESCR and to theprovisions of the Committee on Economic, Social and Cultural Rights (“CESCR”) General CommentNo. 21. Furthermore, the question is approached in an intergenerational perspective: referring to theUnited Nations framework put in place for the enhancement of sustainable development, notablyaccording to the General Assembly United Nations 2030 Agenda and, even more, to the progressivelydeveloping UNESCO norm-set for cultural heritage and sustainability, the work tackles the issue ofthe necessity of preserving and safeguarding from damage and destruction the cultural heritage ofpeoples in order to transmit it intact to future generations, in the name of their right to have access toit as an intrinsic element of their human dignity and well-being.
THE CONSERVATION AND PROMOTION OF THE 'CULTURAL HERITAGE OF MANKIND': WHICH OBLIGATIONS UNDER INTERNATIONAL LAW? / C. Rizzetto ; tutor: N. BOSCHIERO, C. RAGNI, L. COMINELLI ; scientific coordinator: L. SACCONI. Dipartimento di Studi Internazionali, Giuridici e Storico - Politici, 2023. 35. ciclo, Anno Accademico 2022.
KINETICS OF NUCLEIC ACIDS HYBRIDIZATION AND OF COMPLEX DNA STRUCTURES FORMATION ON A BIOSENSING SURFACE
Use of DNA-based molecular probes on a biosensing surface enables unprecedenteddesigns, with nely tuned responsive structures. Furthermore, cost and productionof DNA, in addition to its overall ability to interact with various biological molecules,make DNA the ideal candidate for biosensing surface functionalization. However, allDNA based biosensing relies at some point on the pairing of to single stranded DNAmolecules, namely DNA hybridization. I utilized a label-free, optical, multiplexed,biosensing platform to study probing capabilities of surface grafted DNA, from simpleshort sequences to highly ordered complex nanostructures. My PhD researchstarted with the study of kinetics of simple oligo hybridization on surface. DNA hybridizationon surface is usually treated under the well established Langmuir model,used to treat majority of surface adsorbtion phenomena. However, electrostatics ofDNA phosphate backbone in addition to high denstiy of DNA monolayer on sensingsurface, prevents this model to be applied in all it's potency. We observed strongsuppression in binding kinetics even at concentrations below the KD. Moreover,this suppression correlated positively with DNA probe density, indicating a possibleelectrostatic inuence. Based on the electrostatic theory of DNA monolayers Ideveloped a simple model accounting for the electrostatic penalty associated withentry of DNA molecule into charged DNA monolayer. Moreover, in addition to electrostaticrepulsion hampering, steric eects arising from such high density areas alsoaect the overall sensitivity and rapidity of surface nucleic acids sensing with DNA.We proposed dierent strategies to combat this obstacles, including varying saltconcentrations to counter electrostatic repulsion, grafting DNA probes on hydrogel,eectively reducing charge density. Use of multiple strands was also proposed consideringthe observed high anity towards DNA hybridization to partially doublestranded probes, that provide further stacking stabilization, and decrease the dissociationevents. DNA as a sensing probe was also successfully utilized in sensingmicro-RNA (miRNA), a widely acknowledged and used biomarker for early-diseasediagnostics. MiRNA molecules, like any other RNA molecule, readily hybridizeswith DNA, forming a RNA/DNA hybrid, however, miRNAs specically are veryscarcely and non-uniformly distributed in sera, blood and tissue, this inhibits theavailability of miRNAs, despite their potential as biomarkers. We grafted variousDNA strands with complementary sequences to 5 dierent known miRNAs. Ourmultiplexed assay was able to detect as low as 0.5 pM miRNA while yielding 30xfold mass amplication 90 minutes after the sample injection. Amplication wasachieved by a specic antibody (Ab1) targeting DNA-RNA hybrids, polyclonal secondaryantibody (Ab2) targeting primary antibody and careful system optimization.A simple numeric model was developed accounting the formation of DNA/RNA hybrids,Ab1 binding to hybrid and formation of Ab1-Ab2. Last interaction was treatedas a competitive system, since the Ab pairs also tend to form in solution as well as on surface. System was optimized for amplication factor while keeping the miRNAconcentration and total assay duration xed. The amplication signal was shown tobe sequence dependent, since the kinetics of DNA/RNA hybridization depends onthe sequence and predictive models are not available. DNA microarrays are oftenemployed to study the binding and kinetics of various transcription factors. Weinvestigated binding yeast gene regulator Gal4, on spots containing consensus andnon-consensus sequence both in the form of simple DNA strand and DNA hairpinstructure. Through the experimental observations, we found that the initial bindingstep is charge mediated and can be therefore ne tuned through ionic conditions.A Two-step nested well model was built, to explain these observations
KINETICS OF NUCLEIC ACIDS HYBRIDIZATION AND OF COMPLEX DNA STRUCTURES FORMATION ON A BIOSENSING SURFACE / L. Vanjur ; supervisor: M. Buscaglia ; director of the school: M. Paris. Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, 2021 Jan 26. 33. ciclo, Anno Accademico 2020. [10.13130/vanjur-luka_phd2021-01-26].
THE COVARIANT APPROACH TO SPACE TIMES ANDSOME EXTENDED THEORIES OF GRAVITY
The thesis deals with the covariant description of the geometry of some fundamental spacetimes and their realizations in two recent extensions of GR: Cotton and conformal Killing gravity. In Generalized Robertson - Walker spacetimes the Ricci tensor is the sum of a perfect fluid term and an anisotropic term given by the electric component of the Weyl tensor. In standard cosmology the anisotropy vanishes: this allows to specify the geometric correction due to the extended theory as a perfect fluid.The analysis is then extended to the broader class of doubly-warped spacetimes, employed in investigations of radiating stars and cosmological models. Here the Ricci tensor also contains a heat flow and anisotropic terms originating from the presence of a spatial acceleration vector. The geometric corrections in extended theories are investigated.In spherically symmetric static spacetimes the Faraday tensor is explained. General Relativity and extended theories coupled to linear or non-linear electrodynamics are explored.Cotton gravity was proposed to explain the shape of galaxy rotation curves without imposing dark matter. From the geometric point of view it is shown that the field equations can be recast into Einstein's ones augmented by an arbitrary Codazzi tensor. This simplifies the search of solutions.A new cosmological theory theory was proposed by J. Harada in 2023 to describe the effects of the presence of dark energy without assuming it. Remarkably, the field equations may be recast into Einstein equations augmented by an arbitrary divergence-free conformal Killing tensor, whereby the name conformal Killing gravity. With the new Friedmann equations the growth of the density contrast in a matter-dominated Universe is found to be very close to the \Lambda CDM behavior. The fit of the theoretical Hubble parameter H(z) with only Cosmic Chronometers or combined with Barion Acoustic Oscillations datasets, indicates the occurrence of a Big Rip or an exotic transition.
THE COVARIANT APPROACH TO SPACE TIMES ANDSOME EXTENDED THEORIES OF GRAVITY / C.a. Mantica ; supervisor: L. G. Molinari ; director of the school: A. Mennella. - Dipartimento di Fisica, Via Celoria 16, Milano. Dipartimento di Fisica Aldo Pontremoli, 2024 Nov 18. 37. ciclo, Anno Accademico 2023/2024.
IL CONTRATTO DI ASSICURAZIONE IN GIAPPONE TRA TRADIZIONE, INNOVAZIONE E LEGAL TRANSPLANT
This thesis aims to examine the regulation of insurance contracts in Japan as amended following the introduction of the Insurance Act of 2008. The study of this reform, embedded within the context of a fundamental process of modernization of the country's Private Law, provides significant insights into framing the general principles underlying Japanese Contract Law, the relationship between negotiation autonomy, technical-structural elements of insurance, and consumer’s rights protection, as well as the role played by courts in this field of law. Building upon numerous comparative studies focusing on Japan, the work begins with a review of the state of the art, particularly addressing the general characteristics of the system, comparative issues, and key taxonomic classifications. Having defined the methodology, the research proceeds with an analysis of the sources of Japanese insurance contract law, with particular attention to the interaction between positive law and general principles, and the innovative role played by the judiciary in this matter’s development. Subsequently, the regulation of insurance contracts will be analytically examined, taking into account how case law, doctrine, and contractual practice supplement and complement sector-specific legislation. Finally, considering the significant integration of Artificial Intelligence systems in the creation, underwriting, and management of insurance contracts in recent years, some remarks will be devoted to the Japanese approach to the governance of Artificial Intelligence systems and Insurtech.
IL CONTRATTO DI ASSICURAZIONE IN GIAPPONE TRA TRADIZIONE, INNOVAZIONE E LEGAL TRANSPLANT / D.l. Totaro ; tutor: R. E. Cerchia ; cotutor: A. Candian ; coordinatore: G. Ludovico. Dipartimento di Scienze Giuridiche Cesare Beccaria, 2024 Jul 03. 36. ciclo, Anno Accademico 2023/2024.
FORENSIC TOXICOLOGY IN UNCONVENTIONAL BIOLOGICAL MATRICES: THEEMERGING ARCHAEOTOXICOLOGICAL FIELD APPLIED TO BONE REMAINS ANDBRAIN TISSUE IN ANCIENT POPULATIONS
Forensic toxicology is a specialized branch of toxicology that plays a critical role in forensic scienceby detecting and interpreting the presence of exogenous substances within biological matrices formedico-legal purposes. The field involves both qualitative and quantitative analyses, wherequalitative analysis confirms the presence of specific molecules, and quantitative analysis assessestheir concentrations to determine their possible role in forensic cases. For example, theconcentration of a drug might be evaluated to ascertain if it contributed to a lethal intoxication.Forensic toxicological analyses traditionally rely on conventional biological matrices, such as blood,urine, and visceral samples. However, these matrices may not always be available, especially in casesinvolving advanced decomposition or skeletonization. In such situations, forensic toxicologistsfocus onto unconventional matrices, which include bones, nails, hair, teeth, and in certain caseseven paraffin-embedded tissues and fabric remains. Moreover, as one of the most durable biological matrices, bones can retain molecules oftoxicological interest long after other tissues have decomposed. This characteristic makes bonetissue an invaluable resource in cases where skeletal remains are the only materials available foranalyses. Despite its potential, bone toxicology is less studied and understood compared to othermatrices, with significant challenges in both extraction techniques and data interpretation.However, even if the extraordinary potential of the bone matrix is known, limited information isavailable in the literature on the use of unconventional biological matrices such as bone tissue.Similarly, less is known about how to prepare the bone tissue and how to extract and interpret thexenobiotics that can be extrapolated from the bone matrix.To overcome the lack of information regarding the bone matrix, the primary aim of this Ph.D.project was focused on the detection of analytes of toxicological interest in both forensic andarchaeological human skeletal remains trying to find a proper and efficient bone extractionprocedure to maximize the toxicological investigation and data interpretation. After themaximization of the extraction power of a method, bone toxicology was sided by bone microscopyto better understand how and how much the molecules can remain entrapped inside the bonetissue and what can influence their detection in this substrate. Thus, a secondary objective of theproject was to evaluate if the microstructural changes or microtaphonomic influences (i.e., theprocesses affecting remains post-mortem) might affect the toxicological results with the aim tofurther interpret the toxicological results yet maximized by a new and effective extractiontechnique.Thus, as forensic toxicology continues to evolve (with new extraction techniques and bone matrixinterpretation) and its integration with other disciplines, such as forensic anthropology, it promisesto enhance our ability to reconstruct the lives and identities of individuals from skeletal remains,whether in contemporary forensic cases or historical contexts. Moreover, the emerging field ofarchaeotoxicology further broadens the scope of forensic toxicology, applying its techniques toancient remains to uncover historical medical practices and social habits. The Ca’ Granda project,the third aim of this project, exemplifies the potential of this interdisciplinary approach, offering awindow into the lives of the Milanese individuals of the Modern Age (1600s) and underscoring theimportance of forensic toxicology in both contemporary and historical investigations.Finally, the fourth and last objective was to expand the traditional biological profile of skeletonizedindividuals that can be reconstructed with anthropological investigation integrating thetoxicological profile of a
FORENSIC TOXICOLOGY IN UNCONVENTIONAL BIOLOGICAL MATRICES: THE EMERGING ARCHAEOTOXICOLOGICAL FIELD APPLIED TO BONE REMAINS AND BRAIN TISSUE IN ANCIENT POPULATIONS / G. Giordano ; tutor primo e secondo anno: F. Sardanelli ; tutor terzo anno: F. Secchi ; co- tutor: C. Cattaneo ; supervisore: D. Di Candia ; coordinator: C. Sforza. Dipartimento di Scienze Biomediche per la Salute, 2024 Jan. 37. ciclo, Anno Accademico 2023/2024.
CALCAREOUS NANNOFOSSIL BIOSTRATIGRAPHY AND PALEOECOLOGY OF THE LATE TURONIAN TO EARLY CAMPANIAN INTERVAL AND OF OCEANIC ANOXIC EVENT 3.
The Late Cretaceous was punctuated by several periods of global perturbations of the climate-ocean system that lead to widespread organic carbon-rich marine black shale deposition, termed Oceanic Anoxic Events (OAEs). The OAE3 represents the last episode of anoxia dated as Coniacian-Santonian. Compared to previous anoxic events, OAE3 has a regional extension, rather than a global significance, since it is confined to the equatorial-south Atlantic Ocean and adjacent basins (e.g. Caribbean Basin,Western Interior Basin). Another major difference of OAE3 relative to previous OAEs resides in the absence of a prominent δ13C anomaly, but the occurrence of several minor positive and negative excursions in the Coniacian-Santonian interval. Moreover, the deposition of organic carbon-rich sediments (black shales) seems to be diachronous even in the central-south Atlantic Ocean.This PhD project focused on calcareous nannofossil biostratigraphy and paleoceanography of the Late Turonian to Early Campanian time interval (Late Cretaceous), and specifically of the Coniacian-Santonian. Nannofossil assemblages were investigated semiquantitatively and quantitatively to obtain a detailed biostratigraphic framework for assessing the paleoenvironmental changes across OAE3. The study was performed on Deep Sea Drilling Project (DSDP) and Ocean Drilling Program (ODP) sites and on-land sections, situated in the equatorial and south Atlantic Ocean, the Indian Ocean and theAnglo-Paris Basin. The results of my investigation were integrated with data from literature in order to achieve a solid framework of events for the Coniacian-Santonian time interval and provide a paleoceanographic characterization of OAE3. A detailed revision of the taxonomy was accomplished for solving a few taxonomic incongruities and obtain a unified updated nomenclature. This was the basis for the coherent high-resolution biostratigraphy of the selected sites, in order to establish dating and correlations at supra-regional scale. The four standards biozonation schemes available for the Upper Cretaceous were applied to attain the maximum resolution and test their individual applicability in different oceanic basins and oceanographic settings. The major objective of this PhD thesis was the reconstruction of presumed paleoceanographic changes across OAE3 as recorded by abundance and composition of calcareous nannofossil assemblages. Quantitative analyses of calcareous nannofossil assemblages allowed the characterization of paleotemperature and nutrient changes prior, during and after OAE3. Nannofossil-based paleotemperature obtained in this study and documented in the literature do not show significant changes in the studied successions, perhaps due to unresolved unambiguous temperature-related taxa in the Late Cretaceous and/or minor (subtle) temperature changes across OAE3. As far as paleofertility isconcerned, the nannofossil taxa indicative of higher nutrient content display very different patterns atthe various sites, suggesting that OAE3 was not characterized by a global fertilization episode. Indeed, fertility remained globally low with the exception of upwelling areas characterized by meso-eutrophic conditions also in the intervals preceding and following OAE3. The quantitative analyses conducted in this study pointed out relatively large fluctuations in abundance of genera Micula and Marthasterites in the Coniacian-Santonian interval. These abundance peaks could be correlated with analogous events described in the literature, even if with different values of abundance at the various sites/sections. Two Marthasterites (M. furcatus) abundance peaks and five Micula (M. staurophora) abundance peaks were distinguished across OAE3. The paleoecological affinity of Micula,and specifically M. staurophora, as well as that of genus Marthasterites remains unexplained, so the causes of their abundance peaks continue to be elusive
CALCAREOUS NANNOFOSSIL BIOSTRATIGRAPHY AND PALEOECOLOGY OF THE LATE TURONIAN TO EARLY CAMPANIAN INTERVAL AND OF OCEANIC ANOXIC EVENT 3 / F. Miniati ; tutor: E. Erba ; coordinatore F. Camara Artigas. Università degli Studi di Milano, 2021 Mar 25. 33. ciclo, Anno Accademico 2020.
PLASMA JNK3 AS A NOVEL PERIPHERAL BIOMARKER IN ALZHEIMER'S AND PARKINSON'S DISEASES: EARLY DIAGNOSIS AND DISEASE MONITORING
Alzheimer’s (AD) and Parkinson’s diseases (PD) are neurodegenerative disorders that share overlapping molecular mechanisms but still lack accessible, disease-specific peripheral biomarkers for early diagnosis and disease monitoring. This thesis investigated the potential of c-Jun N-terminal kinase 3 (JNK3), a neuron-specific stress-activated kinase, as a novel peripheral biomarker with translational value in both AD and PD.In the AD cohort, which included patients with AD, mild cognitive impairment (MCI), and controls (CTR), JNK3 levels were measured in both cerebrospinal fluid (CSF) and plasma. Plasmatic JNK3 levels were significantly increased in MCI patients compared to both AD patients and controls, suggesting that JNK3 may reflect early synaptic dysfunction preceding overt neurodegeneration. In CSF, it was observed higher JNK3 levels in AD compared to controls. Notably, JNK3 levels in CSF showed an inverse correlation with age in both MCI and AD groups, supporting its role as a marker of early neuronal stress rather than aging.Instead, only plasma samples were analysed in the PD cohort, which included individuals with isolated REM sleep behaviour disorder (iRBD), de novo PD, late-stage PD, and healthy controls (HC). Plasmatic JNK3 levels progressively increased from iRBD through to late-stage PD and were significantly associated with disease severity and the presence of peripheral phosphorylated α-synuclein in skin biopsies. JNK3 levels showed a significant inverse correlation with motor symptom severity and were more sensitive than neurofilament light chain (NfL) in identifying early disease stages. Unlike NfL, JNK3 did not correlate with age in pathological groups, further supporting its specificity.Importantly, JNK3 was quantified using a commercially ELISA kit, demonstrating its scalability, low invasiveness, and cost-effectiveness for potential clinical use.In conclusion, this study identified plasma JNK3 as a peripheral biomarker in both AD and PD. Its ability to detect early pathological changes, particularly in prodromal stages MCI and iRBD, respectively in AD and PD supports its potential application in early diagnosis, patient stratification, and longitudinal disease monitoring in both clinical and research settings.
PLASMA JNK3 AS A NOVEL PERIPHERAL BIOMARKER IN ALZHEIMER'S AND PARKINSON'S DISEASES: EARLY DIAGNOSIS AND DISEASE MONITORING / A. Giani ; tutor: T. Borsello; coordinatore: G. D. Norata. Dipartimento di Scienze Farmacologiche e Biomolecolari Rodolfo Paoletti, 2025 Dec 15. 38. ciclo, Anno Accademico 2024/2025.
PATHOPHYSIOLOGY OF VON WILLEBRAND FACTOR IN BLEEDING AND THROMBOSIS
von Willebrand factor (VWF) is a multimeric glycoprotein mainly known to be involved in primary hemostasis recruiting platelets at the site of damaged vessels and acting as factor VIII (FVIII) carrier.Quantitative or qualitative alteration of VWF protein is responsible for von Willebrand disease (VWD). Conversely, increased VWF levels have been found to be associated to different thrombotic disorders such as arterial thrombosis, ischemic stroke and venous thromboembolisms. The dual role of VWF in both bleeding and thrombosis, with a focus on VWD and deep vein thrombosis (DVT), are discussed in this thesis. In the first part, we described the approach used to perform VWD diagnosis, including the use of in silico tools and heterologous cell systems to confirm the disease-causing role of VWF variants.In Chapter 2, we described the approach used to perform a differential diagnosis between type 2A and 2B VWD in a pediatric patient carrying a de novo and novel variant. The biochemical characterization allowed us to exclude type 2M due to a loss of high molecular weight multimers and reduced VWF:CB. The finding of a novel deletion, despite its localization in the A2 domain, did not allow for discrimination between type 2A and 2B VWD. The conventional approach would require to perform the ristocetin-induced platelet aggregation (RIPA) to discriminate type 2B VWD from the other type 2. This assay needs a relatively large amount of fresh blood, which is especially undesirable in the case of a pediatric patient. Therefore, we opted for an alternative approach using a platelet-dependent VWF activity (VWF:GPIbM) ELISA. This assay is able to discriminate type 2B VWD patients from the other type 2 VWD patients and it can be done using a small amount of frozen plasma sample. This led us to confirm that our patient was affected by type 2A VWD. In Chapters 3 and 4, we combined the use of predictive in silico tools and heterologous cell systems to prove the pathogenic role of the VWF variants identified in our VWD patients. In both chapters, in vitro expression studies have been performed by transient transfection of wild type and mutant expression vector into HEK293 cells. In Chapter 3, we evaluated the effect of the previously reported type 1 variant p.Arg1379Cys, identified in five unrelated patients. Of them, one was diagnosed as affected with type 1 VWD, whereas the other four had a type 2M diagnosis. These latter patients also carried a polymorphism p.Ala1377Val, which role has been initially underestimated. The in silico evaluation showed that both variants destabilize the A1 domain, leading us to hypothesize a synergistic effect resulting in a decreased capacity of VWF to bind GPIb. Then, we tested the capacity of the wild-type, mutant, and hybrid recombinant (r) VWF to bind recombinant glycoprotein Ibα (rGpIbα) in presence of an increasing concentration of ristocetin. In this way, we were able to confirm that the synergistic effect exerted by these two variants was responsible for patients’ type 2M phenotype. Chapter 4 described the characterization of two unrelated Italian patients diagnosed as affected by type 1 VWD. Both were found to be heterozygous carriers for the same novel variant p.Thr274Pro localized in the VWF propeptide (VWFpp). This variant showed a dominant negative effect in contrast with other previously reported variants found nearby it, which have been described to be responsible for type 3 or type 2A/IIC VWD.Therefore, we decided to perform an in vitro study including immunofluorescence with the aim to elucidate the disease-causing mechanism. Our results showed that the p.Thr274Pro was responsible for patients’ phenotypes through a combined mechanism including defective synthesis, secretion and an impaired multimerization process.The second part of this thesis focused on the characterization of type 3 VWD, the rarest and most severe manifestation of this
PATHOPHYSIOLOGY OF VON WILLEBRAND FACTOR IN BLEEDING AND THROMBOSIS / M.t. Pagliari ; supervisore Università degli Studi di Milano: F. Payvandi ; supervisori Leiden University Medical Center: H. Eikenboom, F.R. Rosendaal ; coordinatore dottorato in Medicina Traslazionale: C. Sforza. Dipartimento di Fisiopatologia Medico-Chirurgica e dei Trapianti, 2023 Sep 20. 35. ciclo, Anno Accademico 2022.
DIABETIC ENCEPHALOPATHY AND NEUROACTIVE STEROIDS: OBSERVATIONS IN FEMALE RATS
Diabetes mellitus (DM), a chronic metabolic disorder, may induce neurophysiological and structural changes in the central nervous system (i.e.,diabetic encephalopathy). Diabetic encephalopathy (DE) is one of the most common and severe chronic complications of DM, characterized by impaired cognitive and memory functions, and structural, neurochemical, and electrophysiological abnormalities. These alterations are associated with decline in cognitive processes, increased risk of cerebrovascular and Alzheimer’s disease, dementia, and psychiatric disorders.Memory and cognitive impairments are associated with hippocampal dysfunction, but the mechanisms behind these alterations are not fully known. Based on data collected in literature, the development of cognitive deficit seems to be the result of the concomitant presence of different processes such as mitochondrial dysfunction, oxidative stress, neuroinflammation and aberrant synaptogenesis in several brain areas of male rat model of type 1 DM (T1DM). Another important factor involved in the development of DE are steroid molecules because it is widely known that these molecules are regulators of the nervous function. Indeed, our previous data have shown that levels of many neuroactive steroids (i.e., steroids affecting the nervous system functionality) are decreased by three months (long-term) of DM but also by one month (short-term) in male streptozotocin (STZ)- induced rats. Moreover, DE presents differences, between two sexes, in term of incidence, progression and severity of the pathology. However, despite DE shows many sex dimorphic features, no observations in female rats have been so far reported in literature.On this basis, in the present study, firstly, we have explored the impact of one month of diabetes on memory abilities by Novel Object Recognition (NOR) test, on mitochondrial functionality, oxidative stress, synaptogenesis and neuroactive steroids levels in hippocampus and cerebral cortex of female STZ-rats in order to understand whether short-term DM affects these parameters also in females and whether possible changes are different in the two sexes.Data reported in this thesis indicate that short term of T1DM is able to significantly decrease memory abilities. This result is associated with aberrant synaptogenesis (i.e., decrease in the levels of synaptophysin, synapsin and syntaxin) and neuroinflammation (i.e., increased levels of IL-1b and IL-6) in hippocampus of female STZ-rats. Oxidative stress and mitochondrial functionality are not affected in this brain area, possibly due to protective effect of dihydroprogesterone (DHP) and allopregnanolone (ALLO), whose hippocampal levels are significantly increased. These effects are specific for hippocampus since the cerebral cortex does not present a similar increase in steroid levels and consequently shows increased neuroinflammation and oxidative stress possibly due to mitochondria dysfunction.T1DM induced not only a diabetic encephalopathy but also dysbiosis. On this basis, we focused our attention on the possible role of gut microbiota on the development of memory deficits in female STZ-rats in the contest of gut-brain axis. In particular, we explored, whether DM can affect the composition of gut microbiota, the levels of steroids in colon and gut permeability markers. Inaddition, we also evaluated whether correlations may occur among these parameters in female STZ-rats. Results obtained demonstrated that T1DM alters gut β-, but not α-diversity. The pathology is also associated with a significant decrease in pregnenolone (PREG) and increase in ALLO levels in the colon of female STZ-rats. In addition, T1DM altered gut permeability (i.e., decreased of zonulin-1 and claudin-1). Interestingly, we reported a significant correlation of PREG with Blautia, claudin-1 and the NOR index and of ALLO with Parasutterella, Gammaproteobacteria and claudin-1.Altogether, da
DIABETIC ENCEPHALOPATHY AND NEUROACTIVE STEROIDS: OBSERVATIONS IN FEMALE RATS / E. Falvo ; tutor: C. Melcangi ; coordinatore: G. D. Norata. Dipartimento di Scienze Farmacologiche e Biomolecolari, 2023 Apr 05. 35. ciclo, Anno Accademico 2022.
AS HIGH AS EYES CAN SEE: A MODERATE LIBERALISM FOR THE ADMISSIBLE CONTENTS OF PERCEPTION
A philosophically crucial question within contemporary philosophy of perception is to determine what things we can perceive, as opposed to things we can only think about. In this thesis, I defend a “liberal” view of perception which accepts that we can perceive some kinds of high-level contents. I propose an original argument based on describing a relevant psychological mechanism that grants such representational capacity that I call schematization. Schematization describes a process by which perceptual systems (I focus on vision) representationally structure their sensory inputs, prioritizing certain feature dimensions, and implicitly activate (or prime) similar representations stored in perceptual memory. Schematization is a purely perceptual process that allows us to represent contents that are not reducible to low-level contents: aspects. Aspects represent some high-level kind properties of particulars. They represent particulars as having some physical body form that makes them belong to a superficial kind, such as the superficial kind of cat-form or chair-form. Crucially, I argue that aspects cannot represent natural or functional kind properties like cat-hood or chair-hood, since such properties depend on below-surface, non-visible characteristics of objects. I thus argue that careful empirical considerations about the representational capacities of perception vindicate a moderate Liberalism that only admits aspects representing superficial kind properties as the higher-level contents of perception. Aspects are as high as eyes can see.
AS HIGH AS EYES CAN SEE: A MODERATE LIBERALISM FOR THE ADMISSIBLE CONTENTS OF PERCEPTION / Q. Coudray ; tutors : C. Sinigaglia, J. Dokic ; coordinator : F. Guala, J. Dokic, M. Aymes. - Università degli Studi di Milano Statale. Dipartimento di Filosofia Piero Martinetti, 2024 Jul 26. 36. ciclo, Anno Accademico 2023/2024.
'È QUESTA L'ARTE GIOVANE?' MOSTRE, SPAZI, ARTISTI A MILANO TRA GLI ANNI OTTANTA E NOVANTA
This thesis presents an initial study of the Milanese artistic scene between the 1980s and 1990s by conducting a historical-critical analysis of the activities carried out by artists and exhibitions in both public and private spaces. It attempts to answer the question posed by Giancarlo Politi, editor of “Flash Art” magazine, in the opening article of the Young Art Special issue in autumn 1990, where he asks, “But is this young art?”, revealing the challenges in understanding the Italian artistic context. This period represents a delicate transitional moment marked by critics' indeterminacy and the resolute individuality of the artists, as well as a moment of transcending labels deemed inadequate for the complexity of the periodAt the center of this research are the young artists, born roughly between 1955 and 1964, who contribute to the overall renewal of the artistic scene through the timing of their career initiation and the variety of expressive languages they adopted.The thesis operates on two different levels: on one side, it provides an overview to capture the common features and potential contradictions within the identified artistic manifestations, presenting a fluid narrative without resorting to forcing or oversimplifications. On the other, it details several group exhibitions held in Milan. These events, involving artists from various parts of Italy, had a significant impact on both the local and national scenes, influencing the debate and development of the period's events. The analysis of these exhibitions serves as a fundamental methodological tool to address critical and artistic issues concurrently and enables an initial engagement with the artworks. This approach allows for the identification of works that gained significant visibility during those years and the artists who were most influential.
'È QUESTA L'ARTE GIOVANE?' MOSTRE, SPAZI, ARTISTI A MILANO TRA GLI ANNI OTTANTA E NOVANTA / G. Zompa ; tutor: S. Bignami ; coordinatore: F. Slavazzi. Università degli Studi di Milano, 2024 Jul 18. 36. ciclo, Anno Accademico 2023.
Chios Mastic Gum from Pistacia lentiscus as a Modulator of Inflammation and Intestinal Barrier Function: An In Vitro and Clinical Investigation
Inflammatory Bowel Disease (IBD) is an immune-mediated disorder characterized by dysregulated inflammation and impaired intestinal barrier function. Although effective, current therapies present limitations, highlighting the need for complementary approaches. Chios Mastic Gum (CMG), a resin from Pistacia lentiscus, has shown anti-inflammatory and barrier-protective properties in preclinical models. During this PhD project, an integrated clinical and mechanistic study was conducted to evaluate CMG (trade name Selpic) in patients with IBD.A randomized, double-blind, placebo-controlled trial involved 31 adults diagnosed with ulcerative colitis or Crohn's disease who were in clinical remission yet exhibited subclinical inflammation. Standardized questionnaires and faecal biomarkers (calprotectin, zonulin), were utilized to assess clinical outcomes. Concurrently, in vitro experiments compared Selpic with its isolated Supermastic® triterpene fraction in Caco-2 cells and lipopolysaccharide-stimulated THP-1 macrophages. Clinically, no significant differences were observed in the questionnaires and calprotectin levels, although a downward trend was noted for the latter (p=0.062). Faecal zonulin levels showed a significant decrease over time (p=0.017), with within-group analysis indicating a significant reduction in the Selpic group (p=0.036). No intergroup treatment effect was identified (p=0.107). In vitro, Selpic was found to suppress pro-inflammatory cytokine levels, diminish reactive oxygen species, encourage anti-inflammatory macrophage polarization, and enhance the expression of ZO-1 tight junction protein. Still, it modulated the endocannabinoid system by reducing N-acylethanolamines SEA and PEA. Analysis of gut microbiota indicated no alterations in α-diversity, but a temporal shift in β-diversity was observed within the Selpic group.In conclusion, faecal zonulin levels decreased over time, with a significant reduction in the Selpic group, though no intergroup treatment effect was confirmed. These results, combined with in vitro immunomodulatory and barrier-restorative effects, support Selpic potential as an adjuvant nutraceutical in IBD management. Larger multicentre trials are warranted to validate its clinical utility.
Chios Mastic Gum from Pistacia lentiscus as a Modulator of Inflammation and Intestinal Barrier Function: An In Vitro and Clinical Investigation / O. Xynomilakis ; tutor: P. Ciuffreda ; cotutor: R. Ottria ; coordinator: F. Cheli. Dipartimento di Scienze Biomediche e Cliniche, 2025 Jan 26. 38. ciclo, Anno Accademico 2024/2025.
ASPECTS OF DATA STRUCTURE IN MACHINE LEARNING
"It is widely believed that understanding data structure is a crucial ingredient to push forward our comprehension on how (and why) modern machine learning works.Still, most of the theoretical results we have are obtained under very simplifying assumptions on the structure of the training data.In this Thesis, I review some novel results on the problem of characterizing the geometric structure of datasets and the consequences that this structure has on learning algorithms.I also provide pedagogical introductions to manifold learning, random geometric graphs theory and supervised binary classification.I focus on three different aspects of the problem.First, I spend some time reviewing techniques to characterize the intrinsic dimensionality of datasets: this is the first ""experimental"" step towards proper theoretical modelling of data.Then, I focus on the problem of finding null models of data in high-dimension: does Euclidean structure survive when the dimensionality of data becomes larger and larger?Finally, I study how geometric data structure alters the expressive potential of simple classifiers."
ASPECTS OF DATA STRUCTURE IN MACHINE LEARNING / V. Erba ; supervisore: S. Caracciolo ; coordinatore: M. Paris. Dipartimento di Fisica Aldo Pontremoli, 2021 Oct 21. 34. ciclo, Anno Accademico 2021. [10.13130/erba-vittorio_phd2021-10-21].
THE ALTERNATIVE SPLICING OF NF-YA REGULATES MOUSE EMBRYONIC STEM CELLS DIFFERENTIATION
Embryogenesis is governed by a fine balance between pluripotency and differentiation controlled by a handful of Transcription Factors (TFs). Some are implied in the maintenance of self-renewal and pluripotency of Embryonic Stem Cells (ESC), others drive cell lineage decisions which give rise to the entire organism. An example of these factors is NF-Y, a heterotrimeric pioneer TF promoting chromatin accessibility to other TFs to regulate gene expression. The regulatory subunit of NF-Y, NF-YA, recognizes and binds the CCAAT box motif, one of the most enriched DNA elements in enhancers and promoters of genes specifically active in ESCs. NF-YA exists in two different isoforms generated by Alternative Splicing of Exon 3. The short (NF-YAs) and the long (NF-YAl) isoforms differ from a stretch of 28-29 amino acids in the N-terminal Q-rich activation domain. Although they display identical DNA-binding properties in vitro, the two isoforms have distinct roles in several biological processes. Regarding embryogenesis, NF-YAs is the predominant isoform in undifferentiated mouse ESCs, being crucial for the regulation of pluripotency and stemness maintenance. By contrast, during Embryoid Bodies (EB) formation process, NF-YAl prevails, suggesting an implication of this specific isoform in the differentiation of mESCs. Our data strengthen the stem-related function of NF-YAs and propose a novel role of NF-YAl in mesoderm formation to shed light on the distinct functions of NF-YA isoforms in the equilibrium between pluripotency and differentiation.
THE ALTERNATIVE SPLICING OF NF-YA REGULATES MOUSE EMBRYONIC STEM CELLS DIFFERENTIATION / S. Polettini ; tutor: D. Dolfini. Dipartimento di Bioscienze, 2025 Apr 17. 37. ciclo, Anno Accademico 2023/2024.
LACTICASEIBACILLUS PARACASEI DSM 34154 (L. CASEI DG®) IN PRECLINICAL AND CLINICAL STUDIES AND ITS GENETIC STABILITY OVER A DECADE OF INDUSTRIAL PRODUCTION
Lacticaseibacillus paracasei DSM 34154, commercially known as L. casei DG® (LC-DG), served as the fil rouge of the whole PhD thesis, which investigates its genetic stability over a decade of industrial biomass production, as well as its effects in preclinical and clinical studies. LC-DG, a well-characterized probiotic strain, has been included in the dietary supplement Enterolactis® on the market for over 20 years because of its proven health benefits. This PhD project initially examined the genetic stability of LC-DG across various industrial production batches through comparative genomic analysis at the single nucleotide level. The findings demonstrate that LC-DG maintains substantial genetic and phenotypic equivalence across all examined lots, effectively mitigating the risk of genetic drift that could compromise probiotic properties. These results highlight the effectiveness of current industrial production practices in preserving the genetic integrity of LC-DG, assuring both producers and consumers that its health benefits are maintained over time, thereby ensuring product reliability. Further preclinical evaluations emphasize LC-DG's ability to effectively survive gastrointestinal transit (GIT) and modulate host immune responses. Then, in vitro studies conducted in a human intestinal cell line model on the AminoAlta™ probiotic formulation (AApf), which combines L. casei DG® with another Lacticaseibacillus paracasei strain, have demonstrated LC-DG's ability to reduce reactive oxygen species (ROS), exhibit anti-inflammatory properties, stimulate macrophage activity, adhere to the differentiated cell layer, and maintain the integrity of the epithelial barrier. These beneficial effects of probiotic supplementation could be crucial in protecting athletes from the physiological stress caused by intense physical activity. The subsequent clinical study (ESA-19) investigated LC-DG's therapeutic potential for treating irritable bowel syndrome (IBS). In this pilot, multicenter, randomized, double-blind, cross-over, placebo-controlled study, the administration of Enterolactis® Ultra significantly influenced the intestinal microbial ecosystem in non-constipated IBS patients, notably by altering short-chain fatty acids (SCFAs) relative proportion, indicating its potential role in alleviating IBS symptoms. This study also highlights the need to identify predictive biomarkers for tailoring personalized probiotic management strategies. Overall, the results of this PhD thesis enhance our understanding of LC-DG, reinforcing the quality, efficacy, and reliability of the probiotic products containing this strain and ensuring that health benefits can be consistently provided to consumers over time.
LACTICASEIBACILLUS PARACASEI DSM 34154 (L. CASEI DG®) IN PRECLINICAL AND CLINICAL STUDIES AND ITS GENETIC STABILITY OVER A DECADE OF INDUSTRIAL PRODUCTION / L. Brunelli ; tutor: S. Guglielmetti ; co-tutor: W. Fiore ; coordinatore: D. Mora. - Università degli Studi di Milano. Università degli Studi di Milano, 2024 Dec 18. 37. ciclo, Anno Accademico 2023/2024.
PEIRCE'S INQUIRY ON HABITS. A CONTEMPORARY PERSPECTIVE
This thesis’ aim is to analysis the concept of habit from the point of view of the philosophy ofAmerican thinker Charles Sanders Peirce (1829-1914). In particular, my goal is to treat habitas a general principle, examining its structure, its “mode of being” and the consequences itentails for cognition and human subjectivity. My thesis analyses and focuses on certain aspectsof Peirce’s vast and complex account of habit, starting from broad ontological questions.Habit appears as a constitutive feature of his categorical system, one of the Americanphilosopher’s most important contribution to the history of thought. From there, movingthematically in the complex architecture of his thought, I highlight the important role habitplays in a pragmatist perspective.My work is inserted in a wider conceptual framework and it is born out of some unsolvedquestion in contemporary secondary literature concerning Peirce and habit. On one hand, I tookinto consideration the history of habit and its theoretical implications. In the last decade wehave witnessed a newfound interest in habits as a fundamental philosophical principle form itsvery beginning, after almost a century of neglect in our discipline. On the other, the pragmatistperspective has become of noteworthy importance in contemporary thought, and the peculiarreading of habit inherent to this tradition is certainly one of its strengths. Peirce plays a peculiarrole in this wider context, and even though his thought on this matter has been somehowthoroughly analysed, it has become necessary to show the richness of his theoreticalpropositions and the contribution that a Peircean philosophy could give to a philosophy ofhabit.Given this context, my guiding insight was to analyse the concept of habit in its broadestpossible sense, in its mode of being. To do so, this work considers two factors. Firstly, it paysspecial attention to accurately frame Peirce’s propositions, isolating some thematic cores Ideem central on which I based my own perspective, and, secondly, highlighting, in theunfolding of the project, the differences, affinities and peculiarities that these thematic coresbring up in relation to the contemporary debate concerning the concept of habit. Peircedevelops a wide and multifaceted concept across the entirety of his thought’s evolution,underpinned by a clear structure: habit is a general principle, triadic and serving as a mediatingfunction, that constructs a tendency to act in a certain way given a certain situation in the future.Taking heed from this general structure, I divided my work in three main sections. In the firstpart, I highlight the philosophical particularity of the concept of habit (in a Peirceanperspective) and I show what vision of the concept of habit I’m going to uphold in myargument, in contrast with some contemporary views of habit. In the second part, theconceptual core of my thesis, I’ll examine the ontological features of habit and I’ll underlineits main metaphysical properties. My work will consider several areas of Peirce’s thought,trying to show a philosophical position that could be confronted with other ontologies of habit,that run through the whole history of this concept, albeit, at times, in disguise. My primary goalis to demonstrate the peculiarity and originality of Peirce’s views on habit, that stems out ofhabit’s secular history and captures some interesting implications entailed since the concept’sinception. Peirce treats habit as a fundamental attribute of being, the central structure of one ofhis categories. My theoretical aim is then to make one step further: habit is not just a categoricalstate, but on the basis of Aristotelian interpretation, it is configured as the middle term betweenthe modes of potency and act, forming a dynamic principle that “enables” from the variouscategories and modes of being,
PEIRCE'S INQUIRY ON HABITS. A CONTEMPORARY PERSPECTIVE / S. Bernardi Della Rosa ; tutor: R. Fabbrichesi ; coordinatore: A. Pinotti. Università degli Studi di Milano, 2022 Sep 30. 34. ciclo, Anno Accademico 2021.
MINING AND LEARNING WEB3 PLATFORMS: A TEMPORAL NETWORK PERSPECTIVE
"In the past decade, there has been a notable shift in Internet services, challenging the traditional structure of Web 2.0 dominated by a few centralized platforms. This transformation has given rise to the concept of Web3, i.e. the design of platforms and software systems built on blockchain technologies to promote a decentralized Web. Indeed, blockchain technology offers many design options for decentralized systems, such as decentralized storage, consensus-based validation of stored data, and the vital option to implement economic systems. Various sectors have witnessed the application of Web3 principles, leading to Decentralized online social media and Decentralized Finance (DeFi), to cite a few well-known examples. Despite the growing influence of Web3, comprehensive studies on its applications, especially beyond decentralized finance, are limited. The vast and timestamped data stored on Web3 platforms offers a unique opportunity for researchers to perform detailed and in-depth analysis of users' traits, potential for the extraction of detailed insights into innovative techno-social systems, characterized by multi-faceted interactions - social, economic, and financial - among people and between people and platform.However, blockchain-based systems are often highly interconnected and exhibit intricate interdependencies that span different interaction layers within the same blockchain and multiple blockchain networks simultaneously. This interconnected yet intricate nature of blockchain-based systems presents substantial challenges, necessitating novel solutions in data collection, modeling, analysis, and prediction tasks. There are many open questions where the key points are the interplay between the temporal and the heterogeneous dimensions characterizing blockchain-based systems. These aspects, still marginal in the current literature, are the main subject of this thesis. This thesis explores the interconnected nature of the Web3 paradigm, utilizing models, algorithms and methods from the field of network science. Network science is a multidisciplinary field, that examines dynamic interactions and complex structures within various system through networks ( also called graphs). In a network, we represent the entities involved as nodes, connected by links (or edges) that represent interactions or dependencies. Modeling through networks has yielded significant insights across domains such as communication networks, urban mobility, scientific collaborations, brain networks, genomics, financial transactions, and social media. Throughout this work, we showcase how network science is a suitable analytics framework to model the complexity of Web3 by tackling different aspects and open questions on Web3 through a network science approach.The thesis is structured as follows: the introduction provides a comprehensive overview of the main research questions, emphasizing the novelty and key contributions. In Part I, ""Modeling Web3 as a network"", the focus is on introducing essential concepts for employing network-based methodologies to model Web3 data, including a background of Web3 platforms and detailing the datasets used. Then, Part II, ""Network evolution dynamics"", delves into the temporal patterns of users' activity and link formation within Web3 systems. After that, Part III, ""The interplay of currency and user behavior"", explores the relationship between user activity and economic dimensions, particularly cryptocurrency and reward systems unique to Web3. And then, Part IV, ""Modeling and prediction of user migration"", investigates cross-chain behaviors like user migration, leveraging heterogeneous temporal data and dynamic socio-economic environments. In addition, Part V, ""Machine learning on multilayer graphs"", introduces a framework enhancing graph neural network performance on the multilayered structure of Web3 platforms. Finally, the Conclusions summarize the key findings, highlighting open problems and suggesti"
MINING AND LEARNING WEB3 PLATFORMS: A TEMPORAL NETWORK PERSPECTIVE / C.t. Ba ; tutor: S. Gaito ; co-relatore: M. Zignani ; coordinatore: R. Sassi. Dipartimento di Informatica Giovanni Degli Antoni, 2023. 36. ciclo, Anno Accademico 2023.
PHARMACOLOGICAL MODULATION OF NEURONAL ACTIVITY FOR THE TREATMENT OF RETT SYNDROME
Rett syndrome (RTT) is a severe neurodevelopmental disorder with an incidence of 1 out of 10000 females born alive. Although considered a rare syndrome, RTT is the most common genetic cause of severe intellectual disability in females worldwide. A typical feature of RTT is the apparently normal early development, followed by a dramatic regression phase that usually occurs 6-18 months after birth. From this moment on, patients begin to present the various classic symptoms of the pathology. However, some recent reports revealed that a series of symptoms, such as hypotonia, jerkiness in limb movements and limited social interactions, are present since early infancy, pointing out that molecular impairment occurs before the onset of clinics symptoms. Nonetheless, their contribution to the pathogenesis is still not understood. More than 95% of classical RTT cases are caused by mutations in the X-linked MECP2 gene and in line with the role of MeCP2 as a master regulator of gene expression, transcriptional maturation is affected in null neurons both in vivo and in vitro, as well as the ability of null neurons to respond to external stimuli. Neuronal activity plays a key role to maintain proper functions of mature neurons, and to ensure their proper development. Accordingly, slight variations from physiological ranges lead to severe consequences, like neurodevelopmental disorders. We thus tested the possible causative link between immaturity and reduced neuronal activity by pharmacologically stimulating Mecp2 null neurons within different time windows of brain maturation. Although, most of the pre-clinical studies so far focused on juvenile or adult mouse models of the disease, one of the main purposes of this study was to identify the optimal time window of intervention for the selected therapeutic approach, also testing the value of anticipating the treatment in a pre-symptomatic period to delay the progression of the disease. To trigger neuronal activity, we selected Ampakine drugs that positively modulate AMPA receptors after glutamate binding. Their binding prolongs the flow of current through the channel enhancing synaptic response and LTP. Two different drugs of the class of Ampakines were tested in vivo using the full knock out mouse model of the disease. The efficacy of the treatments on life span and motor and cognitive defects was established. CX546 ameliorated behavioral defects and prolonged lifespan of Mecp2 null mice in translational time window of intervention. However, the drug induced some discomfort in the treated knock out animals. This evidence together with its poor value for clinical trial led us to test a new generation Ampakine (CX1632) that was already proved safe in clinical trials. This new drug, tested in pre-symptomatic and early symptomatic time windows of intervention, confirmed its total safety in vivo and, more importantly, demonstrated beneficial effects at both the behavioral and molecular level in the mouse model of RTT. However, early treatment revealed more powerful in inducing persistent therapeutic effects. Molecular analyses confirmed the activation of AMPAr associated pathways and the recovery of some RTT defects associated to neuronal activity.All in all, our results support the value of an early therapeutic approach acting on neuronal activity as a strategy for the cure of Rett syndrome.
PHARMACOLOGICAL MODULATION OF NEURONAL ACTIVITY FOR THE TREATMENT OF RETT SYNDROME / G. De Rocco ; tutor: N. Landsberger ; director: N. Landsberger. - Milano. Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, 2024 May 09. 36. ciclo, Anno Accademico 2023.
POLITICAL BELIEF SYSTEMS AS NETWORKS OF ATTITUDES: CONCEPTUALIZATIONS AND IMPLICATIONS
This thesis advances the study of political belief systems by integrating network-based approaches from political science, sociology, and psychometrics. The research is grounded in Philip Converse’s seminal work, which conceptualized political belief systems as interconnected networks of attitudes. Despite the foundational importance of this framework, the field has been slow to empirically embrace this networked perspective. This thesis seeks to fill this gap by employing innovative methods to study belief systems. The research is centered around two core questions: First, how can political attitudes be conceptualized as a belief system? This involves refining the construct of a belief system and investigating the implications deriving from considering attitudes as parts of a broader cognitive structure. Second, how can existing network methodologies be integrated to study belief systems? By combining Belief Network Analysis (BNA), Correlational Class Analysis (CCA), and Pairwise Markov Random Fields (PMRFs)—previously isolated within distinct research domains—the thesis promotes cross-fertilization between disciplines. Chapter 2 presents the theoretical and methodological foundations of the belief system literature. Following Converse, I define political belief systems as networks of causally interacting political attitudes. I then discuss why the dominant view on measurement in social sciences — the latent variable model— is limited in studying these constructs. Next, I motivate why network approaches can provide new insights in this field. Finally, I conduct a scoping review of all papers adopting these approaches to study socio-political attitudes. Chapters 3, 4, and 5 present three empirical analyses. Chapter 3 contributes to the growing literature on subjective inequality by applying CCA and Exploratory Graph Analysis (EGA) to survey data from the U.S. and the Netherlands (n = 2,501 and 1,618). These methods uncover distinct cognitive structures in both countries, with CCA identifying groups sharing similar construals and EGA modeling these as belief systems. The analysis shows that the two belief systems of each country have distinct sociodemographic correlates and that attitudes toward inequality are more socially patterned in the U.S. Moreover, we show that belief systems might represent independent factors in determining different levels of support for redistribution. These findings highlight that ignoring the structural organization of attitudes limits our understanding of public opinions on inequality and redistributive policies. Chapter 4 extends the inquiry by examining how multiple political attitudes coalesce to form a political belief system. Using original data from the 2022 Italian general election, this chapter applies three network models to explore variations in belief system structures across different social groups. The results show that political interest increases the interdependence of attitudes, leading to tighter belief systems, while education levels do not have a similar effect. Interestingly, despite ideological differences, voters from Italy’s two main coalitions (left and right) organize their attitudes similarly, whereas Movimento 5 Stelle voters display significantly different belief structures. This chapter addresses a significant gap by showing how distinct political groups organize their support for parties and issues. Chapter 5 introduces a novel approach to Propensity to Vote (PTV) data, conceptualizing PTV as behavioral intentions embedded in a belief system regarding vote choice. Rather than using traditional data reduction techniques, this chapter models PTV data as networks, where items are connected by weighted edges representing their unique shared variance. Using data from the European Election Studies (1989–2019; 28 countries, n = 121,959), this chapter shows that national political contexts influence the structure of PT
POLITICAL BELIEF SYSTEMS AS NETWORKS OF ATTITUDES: CONCEPTUALIZATIONS AND IMPLICATIONS / A. Bertero ; supervisor: C. Vezzoni, co-supervisor: R. Vacca ; co-authors (Chapter 3): G. Franetovic, J. J. B. Mijs Co-Authors (Chapter 5): M. J. Brandt ; coordinator: M. Jessoula. Dipartimento di Scienze Sociali e Politiche, 2025 May 23. 37. ciclo
COMPUTATIONAL METHODS TO STUDY KNOWN AND NOVEL PROTEIN POST-TRANSLATIONAL MODIFICATIONS BY MASS SPECTROMETRY
A post-translational modifications (PTM) is the covalent modification of a protein after its synthesis, by either addition or removal of functional groups. PTMs significantly increase the complexity of the proteome by allowing each protein to exist in different forms, which can have different activity, stability or binding specificity. Because of their central role in protein regulation, PTMs are often deregulated in many diseases, especially cancer.Since all PTMs introduce a change in the proteins mass, one of the leading techniques to study PTMs is Tandem Mass Spectrometry (MS/MS), which can measure the mass of molecules with high precision and resolution. Here, we apply computational methods to MS data, in order to study PTMs from two points of view. In one project, we conducted a comprehensive analysis of Arginine (R) methylation at a global level. To achieve this, we significantly improved hmSEEKER, our in-house developed computational tool for the analysis of MS data from heavy methyl SILAC (hmSILAC) labelled samples, by implementing a machine learning model to identify methyl-peptides with high confidence. The hmSILAC-validated dataset was then combined with SILAC-based quantitative methyl-proteomics data from a set of SILAC experiments in which we profiled R methylation changes in response to different stimuli (e.g. Cisplatin treatment; inhibition of the major R methyltransferases; PRMT1 expression modulation) to generate the ProMetheus database (ProMetheusDB) of high-confidence methyl-sites. The in-depth analysis of R-methyl-sites inside ProMetheusDB reinforced the notion that protein R methylation modulates protein:RNA interactions but also provided new insights, such as the presence of several R-methyl-proteins involved in metabolism and immune response-related pathways or the fact that R methylation correlates differently with S/T-Y phosphorylation in response to different stimuli. Moreover, we employed computational methods to identify a number of protein:protein interactions that could be affect by this PTM and experimentally validated one of them. Finally, to fully exploit the potential of hmSILAC and hmSEEKER, we explored the application of our pipeline to the annotation of unconventional methyl-sites, which are largely uncharacterized.Since MS has emerged as a powerful tool not only to characterize known PTMs but also to discover new ones, in a second project, we tried to expand the annotation of histone PTMs. Histone lysine (K) acetylation and methylation are routinely used identify cancer subtypes and assess their severity; however, the unbiased nature of MS analysis has revealed the existence of several additional modifications that have yet to be systematically studied. Although theoretically possible, using MS to profile all PTMs that can occur on histones is impractical at the moment, due to limitations in the post-acquisition processing of the MS data. As a matter of fact, the most common computational tools for the identification of modified peptides are not suited to search more than 5-6 PTMs at a time, and this impairs the analysis of the combinatorial nature and cross-talk of histone PTMs. To overcome this limitation, we took advantage of a novel peptide search engine named ionbot, which can perform an “open modification search” to identify an arbitrarily large number of modifications. We reasoned that such a tool would be suitable for the detection of hyper-modified histone peptides. Upon filtering of the results, we were able to annotate not only novel histone PTMs, such as short-chain acylations, but also several amino acid substitutions that could have a biological impact on the interactions between histone and other proteins (i.e. DAXX). The analysis presented here permitted us to better understand the extent, dynamicity and biological role of protein R methylation and identified several modifications and mutation events on histone pr
COMPUTATIONAL METHODS TO STUDY KNOWN AND NOVEL PROTEIN POST-TRANSLATIONAL MODIFICATIONS BY MASS SPECTROMETRY / E. Massignani ; added supervisor: G. Pavesi ; internal advisor: M. Pelizzola ; supervisor: T. Bonaldi. Università degli Studi di Milano, 2021 Dec 13. 33. ciclo, Anno Accademico 2021. [10.13130/massignani-enrico_phd2021-12-13].
IDENTIFICATION OF NEW MYC DEPENDENCIES AMONG RNA-BINDING PROTEINS
Increased expression and activity of the MYC protein is a widespread cancerhallmark and renders tumor cells addicted to sustained activation of a variety ofother gene products. Identification of those dependencies can offer newtherapeutic approaches against MYC-driven tumors. Previous studies showed thatRNA processing events have a critical role in MYC-induced tumorigenesis andsurvival. Moreover, we and others observed that multiple genes encoding RNABindingProteins (RBPs) were positively regulated upon MYC activation in variouscell types. Hence, we hypothesized that the activity of specific RBPs could becomerate-limiting for the growth and/or survival of MYC-overexpressing cells. Towardthe identification of those RBPs, we set up high-throughput genetic dropoutscreens, involving both RNAi and CRISPR/Cas9 technologies. We designedlentiviral shRNA and sgRNA libraries targeting 730 RBPs, which we transduced incell lines allowing controlled super-activation of MYC, in order to identify geneswhose expression was specifically required in this condition. A series of candidatesemerged from our screens, including UPF1 and XRN1, two RBPs involved in mRNAturnover and in particular in nonsense-mediated mRNA decay (NMD). Biologicalvalidation in different systems confirmed our screening results and allowed us toextend our observations to other NMD factors, thus identifying NMD as a criticalpathway in MYC-overexpressing cells. Addressing the mechanisms underlying thesynthetic lethality between MYC and NMD shall not only allow us to unravel thisunexpected crosstalk, but shall also pave the way toward the development of newtherapeutic opportunities against MYC-dependent tumors.
IDENTIFICATION OF NEW MYC DEPENDENCIES AMONG RNA-BINDING PROTEINS / C. Spataro ; added supervisor: A. Sabò ; interna advisor: D. Pasini ; phd coordinator: S. Minucci. Università degli Studi di Milano, 2022 Mar 14. 33. ciclo, Anno Accademico 2021.
DIGITALISING HEALTHCARE: A COMPARATIVE ANALYSIS OF THE ITALIAN AND GERMAN SYSTEMS
This works adopts a comparative law approach to analyze the process of healthcare digitalisation in two EU countries, Italy and Germany. The selection of the cases and the legal systems object of the comparison was based on several factors, which respectively relate to a (infra-)structural precondition - the level of overall digitalisation in the two countries - and two institutional dimensions - the territorial structure of the state and the model of healthcare protection. The precondition is that Italy and Germany have a comparable level of overall digitalisation: in fact, a large gap between the two countries would have made a comparison in the field of healthcare digitalisation ineffective. However, along the institutional dimension, the constitutional arrangement of the two countries is different, since Italy is a unitary regional state while Germany is a federal republic. Finally, with respect to the second institutional dimension, the Italian and the German healthcare systems differ substantially, the former presenting aBeveridgean model based on a universalistic national healthcare system, whereas the latter is organized in accordance with the Bismarckian model of social insurance. Both similarities and substantial differences allow to assess whether the development of the digitalisation process follows similar trajectories, or whether it is intimately dependent on the chosen healthcare model, as well as the possible role played by the different constitutional arrangements.Within this framework, the study has several objectives: to identify which digital health tools have been introduced in the two jurisdictions; to identify what obstacles have been encountered in the process of legislative introduction and subsequent implementation; to outline future perspectives. Thus, the purpose is to determine whether there is a common matrix with respect to the challenges that have characterized the process of digitalisation of healthcare which - although benefiting from considerable momentum due to the pandemic - has been underway in both countries for more than a decade, and consequently whether the solutions implemented follow a common trajectory or are on the contrary attributable to specific institutional conditions in the two countries. In addition, since Germany has recently introduced “pioneering” legislation in the area of mHealth, the research sought insights into that legislation for possible future development in Italian legislation. The dissertation consists of four chapters. The first is introductory in nature and it aims to outline both the key concepts needed to understand the characteristics of the two health systems analyzed, and the challenges and opportunities related to digitalisation in health. After a brief consideration of the concept of health and its protection at the supranational and international levels, the main features of a universalistic and an insurance-based healthcare models are outlined. The final part of the chapter is then devoted to the definition of digital health in its various declinations and the benefits it can bring, as well as the challenges it poses to legal systems.The two central chapters are dedicated to the two national systems under comparison and follow a similar structure, being divided into three parts: the first is dedicated to the constitutional principles underlying the right to health and the corresponding obligations of protection on the part of the state; the second is aimed at reconstructing the main characteristics of the health systems, centered on the National Healthcare System (NHS) in the Italian case (SSN) and thebGKV in the German case; finally, the third part is dedicated to digitalisation tools and their implementation. This last part is the core of the research, to which the other two are preparatory and functional. Particular attention is paid in both Chapter II and Chapter III to the Electroni
DIGITALISING HEALTHCARE: A COMPARATIVE ANALYSIS OF THE ITALIAN AND GERMAN SYSTEMS / G. Re Ferre' ; tutor: D. U.Galetta, K. P. Sommermann ; coordinatore: F. Biondi. - Milano. Università degli Studi di Milano, 2024 Jun 27. 36. ciclo
INVESTIGATION OF PLANT DERIVED NANOVESICLES FROM PLANT PRODUCTS AS NEW BIOACTIVE INGREDIENTS FOR FOOD SUPPLEMENTS AND FUNCIONAL FOODS DEVELOPMENT
Plant-derived nanovesicles represent an emerging research area that addresses critical challenges such as the stability, bioavailability, and efficacy of traditionally known plant extracts.This thesis explores plant-derived vesicles obtained from arugula leaves (ALVs) as a novel ingredient for the prevention of metabolic syndrome. Specifically, ALVs were obtained through the development of an innovative production method and their biological activites were investigated both in vitro and in vivo.The results of the in vivo study support the efficacy and safety of plant-derived nanovesicles, highlighting their potential as promising new ingredients for the development of functional foods and food supplements.
INVESTIGATION OF PLANT DERIVED NANOVESICLES FROM PLANT PRODUCTS AS NEW BIOACTIVE INGREDIENTS FOR FOOD SUPPLEMENTS AND FUNCTIONAL FOODS DEVELOPMENT / L. D'adduzio ; tutor: C. Lammi ; phd coordinator: G. Vistoli. - University of Milan, Department of of Pharmaceutical Sciences. Dipartimento di Scienze Farmaceutiche, 2025 May 09. 37. ciclo, Anno Accademico 2023/2024.
SARCOPENIA, LIVER AND CARDIOVASCULAR ALTERATIONS IN A MULTICENTRIC COHORT OF PATIENTS WITH MASLD: EVALUATION OF CLINICAL PRESENTATION AND GENETIC PREDISPOSITION
Background and Aims: Sarcopenia, characterized by the progressive loss of skeletal muscle mass (SMM), strength, and function, is not solely an age-related condition but is also associated with several diseases, including chronic liver disease (CLD) and cardiovascular (CV) disease. Metabolic-dysfunction associated steatotic liver disease (MASLD) is defined by the presence of liver steatosis combined with, at least, one cardiometabolic risk factor, with liver fibrosis serving as a critical determinant of liver-related and CV-related events and mortality. Sarcopenia is highly prevalent in advanced MASLD, although recent studies have highlighted myosteatosis, the accumulation of fat within muscle, as a key driver of liver disease progression. Studies conducted mainly in Asia have reported a high prevalence of sarcopenia even in early MASLD, though it is not routinely assessed. Both sarcopenia and MASLD contribute to atherosclerotic CV damage, especially in the presence of advanced liver fibrosis, and share common pathogenetic mechanisms such as insulin-resistance (IR), low-grade chronic systemic inflammation, mitochondrial dysfunction, and oxidative stress. Additionally, genetic predisposition plays a role in MASLD, with specific polymorphisms in PNPLA3, TM6SF2, MBOAT7, and GCKR promoting severe liver disease, while the HSD17B13 variant appears protective against MASLD. Despite this background, most studies evaluating the relationship between sarcopenia, MASLD, and CV disease are cross-sectional, primarily involving Asian populations, limiting the generalizability of findings and the ability to determine whether the relationship is bidirectional. Furthermore, the absence of standardized criteria for diagnosing sarcopenia in MASLD complicates comparisons across studies, leading to conflicting results. To date, no studies have yet explored the role of genetic variants predisposing to MASLD in the risk of developing sarcopenia and CV damage. This project aims to address these gaps by 1) evaluating the impact of sarcopenia on liver and CV damage in non-cirrhotic MASLD patients, 2) investigating the influence of MASLD-related genetic variants on muscle mass and CV damage, and 3) assessing changes in muscle, liver, and CV parameters over a 5-year follow-up in a subset of patients.Methods: 856 non-cirrhotic MASLD patients were recruited from liver outpatient clinics in Milan, Palermo, and London. At enrollment, anthropometric, demographic, and laboratory data were collected. Liver steatosis was assessed using abdominal ultrasound (US) and controlled attenuation parameter (CAP) measurements via Fibroscan, while liver fibrosis was evaluated using liver stiffness measurement (LSM) at Fibroscan. Severe steatosis was defined as CAP > 280 dB/m, and advanced fibrosis was identified as LSM ≥ 8 kPa. Liver biopsies were performed on 370 patients. Sarcopenia was assessed using bioelectrical impedance analysis (BIA), a non-invasive and radiation-free technique, with the skeletal muscle index (SMI) calculated as SMM/height^2. Patients were divided into gender-specific tertiles based on SMI (lowest, middle, and highest). CV damage was evaluated using carotid doppler US, with carotid intima-media thickness (cIMT) ≥ 0.9 mm or carotid plaques indicating CV damage. In 502 patients, epicardial fat thickness (EFT) was measured via echocardiography, and carotid-femoral pulse wave velocity (cfPWV) via tonometry. EFT ≥ 9.5/7.5 mm in men/women was considered a marker of visceral adiposity, while EFT ≥ 5.2 mm was associated with coronary artery disease. cfPWV > 10 m/sec was used as a marker of arterial stiffness. CV risk was calculated for all patients using SCORE2 and SCORE2-OP algorithms according to European Society of Cardiology (ESC) guidelines, with SCORE2-Diabetes applied to diabetic patients. Genetic polymorphisms in PNPLA3, TM6SF2, MBOAT7, GCKR, and HSD17B13 were analyzed in 437 patients. Finally
SARCOPENIA, LIVER AND CARDIOVASCULAR ALTERATIONS IN A MULTICENTRIC COHORT OF PATIENTS WITH MASLD: EVALUATION OF CLINICAL PRESENTATION AND GENETIC PREDISPOSITION / A. Cespiati ; tutor: A L. Fracanzani ; supervisore: R. Lombardi ; coordinatore: M. Del Fabbro. Dipartimento di Fisiopatologia Medico-Chirurgica e dei Trapianti, 2024 Dec. 37. ciclo
SOCIOLOGICAL DETERMINANTS OF TAX EVASION: AN EXPERIMENTAL ANALYSIS OF THE ITALIAN CASE
How do people behave when their short-term interest of minimizing paying taxes contrasts with thelong-term collective interest in ensuring sufficient tax payments to finance public goods? In this sit uation, individual decision-making is probably affected by factors beyond any material cost-benefitconsiderations, such as cultural, institutional, social, and normative influences. This doctoral thesisaimed to investigate the sociological drivers of individual tax evasion in Italy with a specific focuson the North-South divide. Drawing on an interdisciplinary approach, I developed a theoreticalframework that integrated three classes of non-economic motivations for tax evasion: individual, in stitutional, and social. I argued that when deciding whether to comply with their taxes, people aremotivated by their moral imperatives toward taxation (i.e., individual mechanism), their perceptionof institutional performance in terms of public goods provision (i.e., institutional mechanism), andtheir social norms of tax payment (i.e., social mechanism). I studied these mechanisms using threecomplementary methodologies on various Italian samples recruited online. In a survey, I measuredthe individual, institutional, and social mechanisms to investigate the roots of the North-South taxevasion gap in Italy. Then, to understand why Italians evade taxes so systematically, I studied themain drivers of tax evasion using two experiments where the institutional and social mechanismswere manipulated: a factorial survey, where I measured tax compliance intention given hypotheticalscenarios, and a behavioural experiment, where I measured tax behaviour in an incentivised envi ronment that mimicked the tax system in Italy. Taken together, the results of the three empiricalstudies show that social norms are key to explain tax evasion in Italy. In other words, when decidingwhether to pay their taxes, individuals are partially influenced by the institutional environment,and mostly by their own beliefs about others’ tax payment (i.e., empirical expectations) and theirbeliefs about the social acceptance of tax evasion (i.e., normative expectations).
SOCIOLOGICAL DETERMINANTS OF TAX EVASION: AN EXPERIMENTAL ANALYSIS OF THE ITALIAN CASE / G.l. Pasin ; supervisors: F. Squazzoni, A. Szekely ; Ph.D. programme director: M. Guerci. Dipartimento di Scienze Sociali e Politiche, 2023 Oct 05. 35. ciclo, Anno Accademico 2022.
STRENGTH EVALUATION OF CRYPTOGRAPHIC PRIMITIVES TO LINEAR, DIFFERENTIAL AND ALGEBRAIC ATTACKS.
Cryptanalysis is an effective method for ensuring the security of cryptographic primitives by attacking them with the most advanced techniques.This thesis provides a deep investigation of three different kinds of cryptanalysis for symmetric ciphers, differential, linear and algebraic, applying them to several symmetric ciphers, from the older ones to the most modern. The algebraic approach consists in solving a polynomial system of equations representing a cryptographic primitive and involves a careful choice of the set of key variables to be fixed. The main instruments used to solve the system are Sat solvers and Grobner basis of which a comparison is offered in some cases. Particular focus has been paid to SHA1 hash function and on the stream cipher E0, used in the Bluetooth protocol. Samely, differential and linear cryptanalysis are applied to several symmetric ciphers. In particular, it is shown how to develop in Python an automatic tool for searching differential and linear trails with the constraint programming language Minizinc.
STRENGTH EVALUATION OF CRYPTOGRAPHIC PRIMITIVES TO LINEAR, DIFFERENTIAL AND ALGEBRAIC ATTACKS / S. Polese ; tutor: A. Visconti ; supervisore: A. Visconti ; coordinatore: R. Sassi. Dipartimento di Informatica Giovanni Degli Antoni, 2023 Jan 30. 35. ciclo, Anno Accademico 2022.
DESIGN, SYNTHESIS AND EVALUATION OF ANTAGONISTS TOWARDS BC2L-C
This project aims to antagonize for the first time the superlectin BC2L-C from multi-drug resistant (MDR) pathogen Burkholderia cenocepacia.MDRs such as Burkholderia cenocepacia have become a hazard in the context of healthcareassociated infections, especially for patients admitted with cystic fibrosis or immunocompromising conditions. As other opportunistic Gram-negative bacteria, this pathogen establishes virulence and biofilms through lectin-mediated adhesion. In particular, the superlectin BC2L-C is believed to cross-link human epithelial cells to B. cenocepacia during pulmonary infection.With the ultimate goal of inhibiting the interactions between the N-terminal of BC2L-C and its target human oligosaccharides, we aim to design glycomimetic antagonists. Here we report the structural study of the target BC2L-C-N-terminal by X-ray crystallography, followed by the design and synthesis of a modular fucoside library of C- and N-glycomimetics. Lastly, we report the biophysical evaluation of the generated glycomimetics against BC2L-CNter by techniques such as STD-NMR, SPR, ITC, DSC; resulting in a lead structure with satisfactory affinity and two crystal structures of antagonist/lectin complexes.
DESIGN, SYNTHESIS AND EVALUATION OF ANTAGONISTS TOWARDS BC2L-C / R. Bermeo Malo ; tutor: A. VARROT; tutor: A. BERNARDI ; referee: C. NATIVI; referee: A. TITZ ; coordinator: E. Licandro. Dipartimento di Chimica, 2021 Jun 18. 33. ciclo, Anno Accademico 2020. [10.13130/bermeo-malo-rafael_phd2021-06-18].
EXTEMPORANEOUS PREPARATIONS IN PERSONALIZED THERAPY: THE DESIGN OF ORODISPERSIBLE DOSAGE FORMS.
The advent of printing technologies for the production of orodispersible films (ODF) guides a growing interest in the application of these dosage forms to precision dosing in personalized medicine. Indeed, the tailoring of ODF shape, colour and/or dimension allows end-users to easily identify their own medicinal product, improving both safety and adherence (Chapter 1). At the same time, to open real perspectives towards ODF for personalized dosing, the design of such technologies should advance along with the development of easy and non-destructive assays, based on colorimetry and spectroscopy, which can allow to establish the physical and chemical quality of ODF (Chapter 2).This doctoral thesis aimed to demonstrates the feasibility of a novel printing technology to extemporaneously compound ODF on-demand. The basic idea was to propose a novel apparatus that combines a hot-melt ram extruder with the plate of a 3D-printer. As far as the formulation is concerned, maltodextrins plasticized with glycerol were selected since they are excipients accepted for both children and elderly. The preparation method consists of simple operations, involving the mixing of the drug substance with maltodextrins and other excipients, then the loading of the mixture into the ram extruder, heating, and printing of the single ODF directly on the packaging aluminium foil. The versatility of this technology was tested by loading ODF with drugs having different physicochemical characteristics. First, paracetamol was selected as a model to demonstrate the drug payload which resulted in loading up to 74 mg/ 6 cm2 and, therefore, allowing the preparation of ODF with a drug amount higher than the highest in the market (i.e., 100 mg/ 9cm2) (Chapter 3). Then, diclofenac sodium was loaded as a model of heat-sensitive and bitter drug to prepare ODF intended for the treatment of migraine in paediatric population. The data revealed that, the exposure to relatively low temperature (i.e., approximately 90 °C) during the printing limited the formation of degradation by-products of the drug (< 0.2%). Furthermore, to improve ODF palatability and patients’ handing, a combination of taste-masking agents (TMA), opacifiers, and, when required, an anti-sticking agent are often loaded into ODF. Thus, the effect of these excipients on the physical properties of ODF loaded by diclofenac was also studied. The results revealed that titanium dioxide, selected as an opacifier, improved not only the ODF aesthetic appearance, but also ODF detachment from the primary packaging material, an aspect particularly relevant to prevent breakage during handing (Chapter 4). Olanzapine (OLZ) was finally tested because it can undergo solid-state modifications under different processing conditions. In this case, the comparison on the performance of OLZ ODF prepared by the proposed technology and consolidated solvent casting technique, which requires the use of a large amount of water, revealed that hot-melt ram extrusion prevented the conversion of OLZ from anhydrous Form I to a pseudo-polymorphic form with lower solubility, which could affect the drug bioavailability (Chapter 5). In conclusion, hot-melt ram extrusion printing can be advantageously used to prepare small batches of ODF made of maltodextrins and glycerine, avoiding the use of solvent and harsh temperatures. This basic formula can be exploited to load drugs differing in physicochemical characteristics, and other excipients to provide suitable organoleptic features of the final dosage form.
EXTEMPORANEOUS PREPARATIONS IN PERSONALIZED THERAPY: THE DESIGN OF ORODISPERSIBLE DOSAGE FORMS / G.m. Khalid ; tutor: F. Selmin ; coordinator: G. Aldini. Dipartimento di Scienze Farmaceutiche, 2021 Jan 13. 33. ciclo, Anno Accademico 2020.
CHARACTERIZATION OF EPCAM IN THYROID CANCER BIOLOGY BY THREE-DIMENSIONAL SPHEROIDS IN VITRO MODEL.
Thyroid cancer (TC) is the most common Endocrine malignancy. Currently, the mechanisms responsible for insensitivity to therapeutic treatments, tumor aggressiveness and tumor relapse described in undifferentiated thyroid cancers (UTCs) haven’t been fully elucidated yet. The insurgence of therapy resistance and disease relapse is believed to be due to a subpopulation of cancer cells within the tumor bulk that shows stem-like phenotype and specific tumor-initiating abilities. These cells are called tumor-initiating cells (TICs), and a comprehensive understanding of how to specifically isolate and target them is necessary. In thyroid cancer, TICs have been identified using specific in vitro and in vivo assays, variations of enzymatic activities, expression of well-known stemness markers and expression of membrane markers. Among the possible membrane markers described in literature to identify TICs, Epithelial Cell Adhesion Molecule (EpCAM) has already been defined as a marker for cancer cells with tumor-initiating properties in many solid tumors, where its possible role in tumor progression and aggressiveness has been described. However, this protein is poorly studied in TC and its role in TC pathology, especially in the undifferentiated forms, needs further comprehension. Hence, the aim of my PhD project was to: 1) characterize EpCAM as a putative thyroid TICs marker on poorly differentiated and anaplastic thyroid cancer cell lines, applying both classical monolayer cultures and 3D spheres cultures; 2) establish if the 3D model is a valid in vitro approach to study the expression and role of EpCAM in the context of TC; 3) observe whether TC cell lines respond differently when treated as adherent cells or as 3D spheres. Our data demonstrated that EpCAM is subjected to an intense cleavage process in FRO-derived 3D tumor spheres and that the 3D model is representative of the variability of EpCAM expression and cleavage that we observed in patient-derived tissue samples, with the different gradient of EpCAM cleavages corresponding to different areas of the tumor sections. We also observed that the expression of EpCAM can be modulated by the regulation of its cleavages and that the integrity of the protein seem to be a crucial factor for the initial phase of the generation of 3D spheres in FRO, while the cleavage of the protein may occur in a subsequent phase, induced by the ability of cells to adapt to variations in growth conditions and/or to the microenvironment within the spheres. Moreover, we demonstrated that EpCAM+ and EpCAM- subpopulations respond differently to treatment with a well-known drug commonly used in clinical practice, Vemurafenib (PLX-4032), both in FRO and HTCC3 adherent cells, and EpCAM+ cells appeared to be more resistant. Finally, the 3D sphere model is also a valid in vitro approach to assess cell response to PLX-4032 and we observed that FRO-derived 3D spheres seemed to be more resistant than HTCC3-derived spheres upon PLX-4032 treatment. In conclusion, we believe that EpCAM expression and its cleavage in 3D spheres could play a significant role in putative TC TICs biology and may partially explain the therapeutic failure observed in the more undifferentiated TCs, concomitantly to other mechanisms of resistance and/or escape that may occur.
CHARACTERIZATION OF EPCAM IN THYROID CANCER BIOLOGY BY THREE-DIMENSIONAL SPHEROIDS IN VITRO MODEL / V. Ghiandai ; Tutor: L. Persani; Supervisor: E. S. Grassi; Coordinatore: N. Landsberger; Revisore esterno: E. Puxeddu; Revisore esterno: C. Nucera. Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, 2023 Apr 19. 35. ciclo, Anno Accademico 2022.
THE EFFECT OF TANNINS FROM CASTANEA SATIVA MILL. ON INFLAMMATION INDUCED BY HELICOBACTER PYLORI IN GASTRIC EPITHELIAL CELLS
The eradication therapy against Helicobacter pylori (H. pylori) accounts for the reduction of IL-8 levels in gastric mucosa and the related risk of gastric cancer and ulcer onset. However, the current therapeutic approach has being challenged by antibiotic resistance. The discovery of natural compounds from edible plants with anti-inflammatory and anti-bacterial activities may contribute to the control of the disease and improve the safety profile of eradication. Plants producing tannins, including ellagitannins, have already shown pre-clinical gastroprotective evidences, but pharmacological validations are still required. Accordingly, Castanea sativa Mill. (sweet chestnut) is a well-known source of tannins, traditionally used for gastro-intestinal and respiratory diseases.On the base of the traditional indication, extracts from sweet chestnut are currently allowed as botanical ingredients by several European countries; however, their phytochemical composition and biological activity are poorly investigated, thus limiting the substantiation of health claims.The aim of the phD project was to characterize hydroalcoholic extracts from fruit and leaves of sweet chestnut, with a particular attention to tannin composition. In parallel, extracts were evaluated as potential candidates against gastritis and H. pylori through a multidisciplinary approach, including in vitro pharmacological and microbiological experiments. In the view of nutraceutical application, the work took into account the importance of the botanical origin of the extracts and evaluated the impact of gastric digestion on the stability of natural compounds.The results obtained on the fruit (chestnut) have been published by the research group (Sangiovanni and Piazza et al. 2018): in brief, following a bio-guided approach, hydroalcoholic extracts from chestnut of five different varieties were screened for their ability to impair TNFα-induced IL-8 release in human gastric epithelial cells (AGS). Two varieties (Venegon and Verdesa) emerged for their promising inhibitory activity (IC50s less than 10 μg/mL) and further characterized for their polyphenolic content. HPLC-MS analysis revealed the presence of condensed tannins (oligomeric proanthocyanidins) in extracts from the outer part of chestnut (pericarp and episperm), which correlated with IL-8 impairment (IC50s less than 0.5 μg/mL); on the contrary, the edible part (endosperm) was devoid of proanthocyanidins and inactive. Of note, hydrolysable tannins typical of the bark of sweet chestnut, namely gallotannins and ellagitannins, were absent in chestnut.In line with the literature on tannins, extracts maintained their inhibitory activity after the simulation of gastric digestion. Moreover, the bioactivity was modestly affected by the application of brief thermal treatments (till 100 °C for 30 min), thus suggesting the possibility of recovering chestnut shell for the enrichment of a bioactive chestnut flour. Consequently, an episperm-fortified flour was prepared respecting the original composition of chestnut, then extracted and assayed again for IL-8 impairment (IC50 = 16.35 μg/mL), thus demonstrating the potential application of chestnut by- products in nutraceuticals.Following a similar approach, hydroalcoholic extracts from leaf belonging to the most promising varieties (Venegon and Verdesa) were investigated. Contrarily to the fruit, the ellagitannins castalagin and vescalagin were revealed (1-2%) in leaf, thus prompting further studies on the anti-H. pylori effect.In particular, the antibacterial and anti-inflammatory activity was evaluated in a co-culture model of gastric epithelial cells infected with the bacteria (tumoral AGS in comparison with non-tumoral GES- 1). Leaf extracts inhibited IL-8 release in both the infected cells with IC50s lower than 30 μg/mL andimpaired the bacterial growth at 100 μg/mL. After simulated gastric digestion, these bioac
THE EFFECT OF TANNINS FROM CASTANEA SATIVA MILL. ON INFLAMMATION INDUCED BY HELICOBACTER PYLORI IN GASTRIC EPITHELIAL CELLS / S. Piazza ; tutor: M. Dell'Agli ; coordinatore: A. L. Catapano. Dipartimento di Scienze Farmacologiche e Biomolecolari, 2021 Jan 13. 33. ciclo, Anno Accademico 2020. [10.13130/piazza-stefano_phd2021-01-13].
UNVEILING CARDIAC IL-33/ST2L PATHWAY DEREGULATION IN ANIMAL MODEL OF OBESITY AND CARDIOVASCULAR DISEASE
Obesity has become an increasing public health issue worldwide; it is estimated than over 65% of American adult population is overweight and 35% are actually obese. Criteria for obesity definition is traditionally linked to Body Mass Index (BMI). Although an important correlation exists between BMI and body fat deposit, the BMI criteria cannot actually predict body composition in any given individual due to variability determined by age, sex, race and ethnicity. Besides the weight of an individual, both fat localization and type represent more important estimators of cardiovascular disease. Importantly, the accumulation of visceral fat – more than subcutaneous fat – is linked to cardiovascular disease severity and mortality. Among ectopic fat depots, pericardial and epicardial fat accumulation are intriguingly important. Epicardial adipose tissue (EAT) represents a biological active tissue that secretes adipokines known to promote low-grade inflammation and diabetic vascular complications. The cause is due to the shared blood supply between EAT and myocardium, as indeed EAT provides the 50–70% of the energy used for contraction trough the release of free fatty acids (FFAs). It is intuitive that molecular/metabolic alteration of EAT will be reflected on its secretome and on cardiac metabolism. EAT thickness seems to be associated to ventricular myocardial mass and importantly to myocardial steatosis. A plethora of works highlighted the role of IL-33 and its receptor ST2 within cardiac dysfunction and inflammation associated with obesity. Once secreted in activated or damaged cells, IL-33 modulates the functions of various immune cells through ST2 binding, affecting the proliferation of T cells, macrophages, and innate lymphoid cells. Accordingly, IL-33/ST2 pathway is involved in lipid metabolic diseases and - following its role as immune sensor to infection and stress – it is linked to pro-fibrotic remodeling of myocardium and more in general in cardiovascular diseases. Our group demonstrated in coronary artery disease (CAD) a direct correlation between EAT thickness and IL-33/ST2 signalling imbalance further highlighting a role for EPAC protein involved in cAMP signal transductionIn this PhD Project we aimed to further investigated the IL-33/ST2 effects on cardiac remodeling in obesity, focusing on the molecular pathway that links adipose/cardiac-derived IL-33 to development of fibrosis or hypertrophy. We choose Zucker Fatty rats (ZF) and Zucker Diabetics Fatty rats (ZDF) to overcome the problems linked to tissue availability and to the homogeneity of human EAT samples that need to be collected during cardiac surgery. Since genetic animal models do not fully recapitulate human pathology, we developed in vitro models to mimic adipose and myocardial relationship in vitro. Indeed, we evaluated the effects of visceral adipose tissue (VAT)-derived cellular medium in the modulation of adipose secretome and how they affect myocardial gene expression. Finally, we compared obtained results with those derived from Diet induced obesity (DIO) mice, a naturally occurring model of obesity that reliably resembles human disease. Following both molecular and proteomic analysis, we demonstrated a dysregulation of IL-33/ST2 signalling in both adipose and cardiac tissue, where they affected myocardial gene expression and determined a pro-fibrotic signature. In Zucker rats, pro-fibrotic effects were counteracted by ghrelin-induced IL-33 secretion, whose release influenced transcription factor expression (such as MEF2a), but also increased sST2 and not cardioprotective ST2L form. In this context we observed a reduction of EPAC signalling, that is promoted by VAT secretome and linked to ST2 isoforms balancing regulation. Similar results were obtained in ZDF rats, as both models did not develop evident alteration of cardiac architecture, although we described the increase of the pro-fibrotic signatu
UNVEILING CARDIAC IL-33/ST2L PATHWAY DEREGULATION IN ANIMAL MODEL OF OBESITY AND CARDIOVASCULAR DISEASE / C. Sitzia ; tutor: MM. Corsi-Romanelli; supervisor: E. Vianello ; coordinatore N. Landsberger. Università degli Studi di Milano, 2023 Jan 31. 35. ciclo, Anno Accademico 2022.
MULTIFUNCTIONAL EVALUATION AND VALIDATION OF PRACTICES, TOOLS AND INNOVATIVE MODELS FOR THE LANDSCAPE AND AGROECOLOGICAL MANAGEMENT OF AGRICULTURAL SYSTEMS: TRADE-OFFS AND SYNERGIES IN THE ECOSYSTEM SERVICES FRAMEWORK
BACKGROUND. Severe agrobiodiversity loss trends are ongoing, altering the agroecosystem regulating processes. Biodiversity loss is both a consequence and driver of the current ecological crisis. Among rural systems, it is a pivotal driver of agroecosystems vulnerability and lack of resilience.Multi-spectrum solutions are needed, and this is being more and more recognised among all the different levels of agricultural and environmental studies, guidelines, policies, programs and strategic actions. Several agricultural management options exist: low-intensity and diversified agricultural land use can contribute to reversing these negative trends. Within this context, the agroecological approach specifically seeks the reintegration, within the agricultural systems, of their undermined ecosystem functions. Among the agroecological practices, agroforestry is a recognised, viable, multi-purpose rehabilitating strategy to reduce current environmental externalities of agriculture. To properly address these issues, the monitoring of the effectiveness of mitigation and rehabilitation strategies is needed, and result-based and mixed result-practice based approaches are recommended for securing biodiversity gains.AIMS.Within this framework, this research project focuses on the ecological and agri-environmental sides of farm models validation, based on mixed practice-result-based approaches. The project aims at developing a methodological framework for a multi-functional evaluation and validation of models for the landscape and agroecological management of agroecosystems among specific agri-environmental contexts. It starts from the experience of the farmers network of Polycolturae association, partner of the research project. The Polyculturae farms networking activity is aimed at promoting the agroecological approach through agroforestry-based and nature-based agricultural practices; this aim is also sought through the Biodiversitas label, a tool for ensuring the achieving of high environmental performances of farms, with a specific focus on agrobiodiversity. The Biodiversitas label is currently on a pilot stage and this research project addresses the steps propaedeutic to its final design and implementation.Research activities are directly addressed to two main objectives: Objective 1. Polyculturae model validation:- 1.a. The scientific validation of the agroecological and agroforestry-based model promoted by Polyculturae association- 1.b. The delivering of an analytical methodology to orient farm strategic ecological design and management, optimising the ecological effectiveness of interventions.Objective 2. To orient the further development of the Biodiversitas label, by delivering a proposal on its re-structuring and indicators system integration.METHODS.Research activities are led among the four farms currently member of the Polyculturae association (POLY farms), representing agroecology and agroforestry-based farm models in the western alluvial Po Plain district, Piedmont and Lombardy regions. Objective 1.a is sought through a multi-scale and multi-disciplinary analytical approach based on landscape ecology, floristic and vegetational (phytosociological) analyses and soil organic carbon analyses, coupled to the assessment of agronomic performances (yields and economic performance indicators). Extra-local and local landscape featuring is performed to set context-specific baselines, vulnerabilities and resilience drivers, against which each Polyculturae farm contributions are assessed. From the validation side, POLY farms are compared with local conventional baselines (farm models based on annual crop monocultures with no natural and semi-natural components active management). The Polyculturae model contributions to agrobiodiversity and agroecosystem ecological quality are also assessed from the Ecosystem services perspective, through a land-use based approach, to complement the assessment from a multi-fun
MULTIFUNCTIONAL EVALUATION AND VALIDATION OF PRACTICES, TOOLS AND INNOVATIVE MODELS FOR THE LANDSCAPE AND AGROECOLOGICAL MANAGEMENT OF AGRICULTURAL SYSTEMS: TRADE-OFFS AND SYNERGIES IN THE ECOSYSTEM SERVICES FRAMEWORK / G. Chiaffarelli ; tutor: I. Vagge ; phd coordinator: M. P. M. Guarino. - Milano. Dipartimento di Scienze Agrarie e Ambientali - Produzione, Territorio, Agroenergia, 2025 Mar 26. 37. ciclo, Anno Accademico 2024/2025.
COMPUTATIONAL APPROACHES IN STUDYING INHIBITOR DEVELOPMENT IN HAEMOPHILIA A PATIENTS
Haemophilia A is an X-linked recessive bleeding disorder caused by mutations in the F8 gene, resulting in deficiency or dysfunction of coagulation Factor VIII (FVIII). A major clinical complication in severe cases is the development of neutralizing alloantibodies, known as inhibitors, which undermine treatment efficacy and increase morbidity. This doctoral thesis uses a multidisciplinary approach by combining computational biology, bioinformatics, and high-throughput immunological assays, to investigate the mechanisms of inhibitor development and identify predictive biomarkers for clinical risk stratification.The first study employs a novel IgG epitope mapping method using a random peptide phage-display library in the SIPPET cohort. Mimotope-variation analysis (MVA) was used to characterize FVIII-specific antibody repertoires, and predictive models based on LASSO logistic regression and random forest algorithms achieved strong performance (C-statistics ~0.78–0.80), supporting their utility in pre-treatment risk assessment.The second study explores molecular mimicry by aligning FVIII mimotopes with pathogen-derived B-cell epitopes from the Immune Epitope Database (IEDB). Although some homologies were observed, no significant enrichment was found, providing no support for molecular mimicry as a driver of inhibitor formation.The third study involves genome-wide DNA methylation analysis to identify epigenetic differences associated with inhibitor risk. Differentially methylated CpG sites were found in genes linked to immune regulation and tolerance, suggesting epigenetic contributions to FVIII immunogenicity.Overall, this thesis demonstrates the value of integrative computational approaches in elucidating immune responses in Haemophilia A and contributes to the development of predictive tools and personalized treatment strategies in translational medicine.
COMPUTATIONAL APPROACHES IN STUDYING INHIBITOR DEVELOPMENT IN HAEMOPHILIA A PATIENTS / H. Chand ; tutor: F. Peyvandi ; co-tutor: R. Palla ;coordinator: M. Clerici. Dipartimento di Fisiopatologia Medico-Chirurgica e dei Trapianti, 2025 Jun 18. 37. ciclo, Anno Accademico 2024/2025.
INHIBITING THE NUCLEAR RECEPTOR ESRRA RESCUES CORE PATHOLOGIES IN CELLULAR AND ANIMAL MODELS OF BATTEN DISEASE
Batten disease (BD), a group of fatal neurodegenerative lysosomal storage disorders, is characterized by vision loss, cognitive decline, and seizures. Our lab recently identified the accumulation of the glycosphingolipid Gb3 as a key pathological hallmark in multiple BD subtypes (Soldati et al., 2021). However, the underlying molecular mechanism is unknown. This thesis aims to identify the genes controlling Gb3 accumulation, paving the way for new small-molecule therapies capable of penetrating the central nervous system. Thus, to find genes involved in Gb3 buildup, we performed a high-content imaging screen using a library of approximately 11,000 siRNAs (small interfering RNAs, which silence specific genes) in a human ARPE-19 CLN3 knockout (CLN3KO) cell model. The survey identified 84 candidate genes whose depletion reduced Gb3 levels. We focused on the top candidate hit, Estrogen-related receptor alpha (ESRRA/ESRRA), a nuclear receptor involved in mitochondrial biogenesis, lipid metabolism, and autophagy. Both genetic silencing and pharmacological inhibition of ESRRA with the inverse agonist XCT-790 significantly reduced Gb3 accumulation in both CLN3 and CLN7 disease models, suggesting that ESRRA may represent a novel therapeutic target for multiple BDs. Transcriptomic analysis revealed that pharmacological inhibition of ESRRA with XCT-790 triggers a beneficial metabolic reprogramming by further suppressing defective mitochondrial respiration, activating the PPARG pathway, and inducing the expression of FOX3a, a transcription factor involved in the transcriptional activation of autophagy. Indeed, ESRRA inhibition enhances cellular clearance by boosting autophagic flux by mechanism independent of the nuclear translocation of the master gene of lysosomal biogenesis and autophagy TFEB. To validate these findings in a living organism, we treated CLN7 knockout mice with XCT-790 via dietary supplementation of the compound. The treatment successfully reduced Gb3 and SCMAS accumulation in the brain's cortex and cerebellum, and ameliorated neuroinflammation and motor phenotype.Collectively, this research identifies ESRRA as a novel druggable target for Batten disease. Our findings demonstrate that ESRRA is a novel druggable target and its inhibitor XCT-790 ameliorate disease hallmarks in vitro and in vivo, offering significant therapeutic potential, and a new strategy to restore both lysosomal and mitochondrial function across multiple, distinct subtypes of BDs.
INHIBITING THE NUCLEAR RECEPTOR ESRRA RESCUES CORE PATHOLOGIES IN CELLULAR AND ANIMAL MODELS OF BATTEN DISEASE / M. Aurilia ; tutor: D.L. Medina. Dipartimento di Oncologia ed Emato-Oncologia, 2025 Dec 19. 37. ciclo, Anno Accademico 2024/2025.
FAITHFUL ESTIMATION OF UNCERTAINTIES IN MODERN PDF EXTRACTIONS
The primary focus of this Ph.D. thesis is the precise determination and validation of the uncertainties associated with parton distribution func- tions (PDFs). We introduce and implement the theory covariance method within the NNPDF4.0 framework, enabling the incorporation of theoretical uncertainties into the PDF determination process. Additionally, we revisit and expand upon the closure tests framework, which serves as a tool for val- idating the PDF extraction methodology in a controlled environment. This framework is applied to a dataset that has been deliberately constructed to be inconsistent, allowing for a rigorous assessment of the methodology’s ro- bustness. Furthermore, we utilize this framework to validate the extraction of the strong coupling constant using the correlated replica method. Finally, we present a novel theoretical pipeline, which introduces several technical advancements and underpins all the results discussed in this thesis.
FAITHFUL ESTIMATION OF UNCERTAINTIES IN MODERN PDF EXTRACTIONS / A. Barontini ; supervisore: S. Forte ; coordinatore: A. Mennella. Dipartimento di Fisica Aldo Pontremoli, 2024. 37. ciclo
LE CAUSE DI NON PUNIBILITÀ IN SENSO STRETTO: FONDAMENTO POLITICO-CRIMINALE, CARATTERISTICHE E DISCIPLINA
"The “cause di non punibilità in senso stretto” constitute a category that is the subject of a wide and significant doctrinal debate, due to the various interpretative doubts that arise in relation to the individual institutions attributable to it. The most recent criminal legislation is characterized by a wide use of the “cause di non punibilità in senso stretto”, both in the special part of the Criminal Code and in complementary legislation, and this also in order to ""balance"" the unstoppable phenomenon of expansion of criminal law. In any case, these are problematic institutions because, entailing the waiver of punishment of unlawful and culpable acts, they stand ""in very strong tension with the principle of criminal protection of society that is constitutionally incumbent on the legislature and the state in general"". In the present research work, first of all, a ""mapping"" of the “cause di non punibilità in senso stretto” found in the Criminal Code and complementary legislation was carried out, attempting to identify the rationale behind them. Second, an attempt was made to identify the characteristics that these institutions should have in order to be considered legitimate. Although the provisions that exclude punishability can all be traced back to some extent to the principle of subsidiarity, what emerges from the present analysis is that this “cause di non punibilità in senso stretto” of non-punishability in the strict sense, entailing different consequences in terms of discipline. In any case, in the conclusion of the paper, an attempt has been made to highlight the circumstance that the further one moves away from the perspective of protection of the legal goods presided over through incrimination, the more difficult it is to consider the exclusion of the application of punishment as legitimate in the face of a criminal act."
LE CAUSE DI NON PUNIBILITÀ IN SENSO STRETTO: FONDAMENTO POLITICO-CRIMINALE, CARATTERISTICHE E DISCIPLINA / C. Cataneo ; tutor: A. Della Bella ; coordinatrice: F. Poggi ; revisori: C. Ruga Riva, G. Demuro ; commissari: F. Basile, A. Gargani, A. Gullocommissari: F. Basile, A. Gargani, A. Gullo. Dipartimento di Scienze Giuridiche Cesare Beccaria, 2023 Apr 03. 35. ciclo, Anno Accademico 2022.
GAMMA-PROBE GUIDED SENTINEL LYMPH NODE EXTIRPATION TO ASSESS PATTERNS OF NODAL METASTASIS IN SPONTANEOUS HEAD AND NECK MALIGNANCIES OF THE DOG: A SECOND STEP BASED ON PREVIOUS EXPERIENCES ON MAST-CELL TUMORS
Canine oncology has recently reported interest in the detection of occult nodal metastases, and efforts have been made to identify the best strategy for accurate nodal staging. In this scenario, the present PhD thesis was aimed to improve knowledge on the current understanding of the feasibility and impact of sentinel lymph node (SLN) mapping with radiopharmaceutical and blue dye in cancer-bearing dogs. Initially, we evaluated the technique in canine mast cell tumors (MCT), given the high prevalence of this tumor type, the propensity for lymphatic spread and recognized prognostic impact of nodal metastases. Thirty client-owned dogs with 34 MCT were included. At least one SLN was identified in all but 3 dogs that had a scar from previously excised MCT, and in 30 cases (63%) the SLN did not correspond to the regional lymph node (RLN). As a second step, we explored the impact of SLN mapping and extirpation in dogs with head and neck tumors, given the well-accepted unpredictability of patterns of nodal metastases. Twenty-three dogs with tumors of the head and neck and absence of clinically evident nodal disease (cN0 neck) were prospectively included and underwent tumor excision and SLN extirpation. Reported detection rate was 83%, with at least one SLN identified in all but 4 dogs with thyroid tumors. In 52% of dogs, the SLN did not correspond to the RLN and at histopathology 42% of dogs had nodal metastases, of which 4 differed from the RLN. As the last step of this PhD project, we conducted an explorative study to identify clinical or pathological characteristics that could allow for identification of dogs with MCT at lower risk of SLN metastases, that could be excluded a priori from the SLN mapping and extirpation. Surprisingly, tumor grade was not able to predict the risk of having HN2-3 or HN3 SLN and the only variables that correlated with nodal metastases were tumor size, number or SLN and subcutaneous MCT. Given low to moderate discriminant ability, however, none of these variables could safely determine the exclusion of patients from the procedure. Results of the present project underscore the utility of implementation of SLN biopsy in the management of dogs with MCT and head and neck tumors, given the low correspondence between clinically expected RLN and SLN and the high rate of occult nodal metastases that could be detected with this procedure. Future research should be aimed at investigating the impact on long-term prognosis and disease free interval of the extirpation of metastatic SLN, and identify variables that could allow for exclusion of low-risk patients from the procedure.
GAMMA-PROBE GUIDED SENTINEL LYMPH NODE EXTIRPATION TO ASSESS PATTERNS OF NODAL METASTASIS IN SPONTANEOUS HEAD AND NECK MALIGNANCIES OF THE DOG: A SECOND STEP BASED ON PREVIOUS EXPERIENCES ON MAST-CELL TUMORS / L.e. Chiti ; tutor: D. Stefanello ; internal reviewer: D. D Zane, S. Romussi, A. Belloli ; external reviewer: M. Martano, M.C. Nolff ; coordinator: V. Grieco, F. Ceciliani. Dipartimento di Medicina Veterinaria e Scienze Animali, 2022 Mar 31. 34. ciclo, Anno Accademico 2021. [10.13130/chiti-lavinia-elena_phd2022-03-31].
Multi-organ failure driven by telomere uncapping
The telomere-bound shelterin complex safeguards chromosome ends from being recognized as DNA double-strand breaks, a function mostly dependent on TRF2. Loss of TRF2 results in telomere uncapping, activation of the DNA damage response, and chromosomal end-to-end fusions, with dramatic consequences for genome integrity and cell fate. The impact of TRF2 depletion is context-dependent, causing distinct outcomes in different cell types, however the systemic effects of acute TRF2 loss have remained poorly characterized.In this work, we adopted an inducible Trf2 knockout mouse model to investigate the consequences of telomere uncapping at the organismal level. Trf2 deletion was rapidly achieved across multiple tissues and this event was accompanied by DDR activation and multi-organ functional failure, ultimately leading to lethality within approximately one week. Trf2 knockout elicited a syndrome resembling that occurring upon exposure to high doses of ionizing radiation, including severe bone marrow hypoplasia, leukopenia, hematopoietic stem and progenitor cell dysfunction, and intestinal epithelium degeneration. Bone marrow transplantation failed to extend lifespan in Trf2 knockout mice, whereas it rescued the mortality caused by X-rays at low doses, highlighting the fact that lethality is not primarily dictated by the hematopoietic system, but the consequences of TRF2 loss extend to other organs.Our study demonstrates the essential role of TRF2 in preserving the integrity of the hematopoietic and intestinal systems, presumably owing to their highly proliferative nature, and underscores the catastrophic outcomes of its loss at the organismal level, aligning the effects of telomere uncapping with the pathophysiology of acute radiation syndrome.
Multi-organ failure driven by telomere uncapping / L.a. Trastus ; tutor: F. d'Adda di Fagagna ;co-tutor: V. Costanzo ;internal advisor: S. Campaner ;external advisor: J. Gil PhD coordinator: D. Pasini. Dipartimento di Oncologia ed Emato-Oncologia, 2025 Dec 17. 37. ciclo, Anno Accademico 2024/2025.
THE ASSOCIATION OF CLINICAL PROGNOSTIC FACTORS WITH VENTILATION INHOMOGENEITY: SINGLE CENTRE ANALYSIS OF PATIENTS WITH CYSTIC FIBROSIS FROM 2014 TO 2019
BACKGROUND. Especially in children, spirometry may not capture the underlying lung disease, which has been described in small airways since young ages in cystic fibrosis (CF). Lung Clearance Index (LCI2.5) is one of the derivatives obtained by the multiple breath washout (MBWN2) test, recognized as a sensitive alternative for the assessment of lung function. The potential of this outcome in clinical setting has not been fully explored yet.METHODS. Data were collected for each patient’s visit from 2014 up to September 2019 from the CF Centre of Milano and analyzed according to different perspective. The association between lung function measures and selected prognostic variables was investigated by means of ordinary least-square regression models; disease phenotypes were obtained according to children’s clinical characteristics by means of agglomerative nesting hierarchical clustering; longitudinal association between lung function and selected variables, and phenotypes, was assayed fitting linear-mixed effect models; recurrence of pulmonary exacerbations (PE) was evaluated by extensions of the Cox proportional hazard models.RESULTS. A total of 433 MBWN2 measurements from 245 children with CF were analyzed. Cross-sectionally, both LCI2.5 and FEV1 %predicted showed significant association with CF clinical prognostic variables, i.e., Pseudomonas aeruginosa infection, pancreatic function, BMI Z-score and age, whereas only LCI2.5 was associated with genotype. Two different phenotypes of children around 11 years old emerged, differentiating one from another also by a mean LCI2.5 difference of 3.37 (95%CI: 2.57 to 4.16) and by a mean FEV1 difference of 1.2 (95%CI:0.7 to 1.6) Z-score. Children with the best phenotype were mainly characterized by absence of F508del mutation and by a normal pancreatic function. Longitudinally, LCI2.5 and FEV1 % predicted remained stable over time and showed meaningful differences between phenotypes at each follow-up visit. However, infection by Pseudomonas aeruginosa was associated with longitudinal variation of LCI2.5 only, showing that being free from Pseudomonas aeruginosa lowers LCI2.5 by 0.82 (95%CI: -1.36 to -0.27). Eventually, mean number of PE in the present cohort was 1.8, varying from 0 to 4. Children with higher LCI2.5 are expected to experience a 6% higher risk of PE recurrence during their follow-up (HR 1.06, 95%CI 1.01 to 1.10), whereas FEV1 % predicted did not show any evidence of association. An association with PE recurrence was also detected between phenotypes in favour of children with better clinical status (HR 0.46, 95%CI 0.34 to 0.60).CONCLUSIONS. LCI2.5 shows interesting associations with clinical characteristics not shared with FEV1, %predicted, both in cross-sectional and longitudinal analyses. Clustering has shown a disease profile of children who share negative clinical prognostic factors, also in terms of ventilation inhomogeneity, and are at higher risk of recurrent pulmonary exacerbations. Altogether these findings add to the available literature in confirming the clinical utility of MBWN2.
THE ASSOCIATION OF CLINICAL PROGNOSTIC FACTORS WITH VENTILATION INHOMOGENEITY: SINGLE CENTRE ANALYSIS OF PATIENTS WITH CYSTIC FIBROSIS FROM 2014 TO 2019 / S. Gambazza ; coordinatore: C. La Vecchia ; supervisor: F. Ambrogi. Dipartimento di Scienze Cliniche e di Comunità, 2021 Mar 10. 33. ciclo, Anno Accademico 2020.
THE IMPACT OF VASCULAR SMOOTH MUSCLE CELL SENESCENCE ON MITOCHONDRIA DYSFUNCTION AND ANGIOGENESIS
INTRODUCTION: Atherosclerosis, the principal underlying cause of most cardiovascular diseases, progresses from early fatty streaks to advanced unstable plaques that, if ruptured, cause life-threatening events. A hallmark of plaque instability is intraplaque angiogenesis, the formation of neovessels. These neovessels are often immature and leaky, facilitating intraplaque haemorrhage, which exacerbates inflammation, promotes plaque growth, and culminates in rupture and thrombotic complications. While hypoxia is a critical driver of this process, additional pro-angiogenic mechanisms within the plaque microenvironment remain incompletely defined. Atherosclerosis is also an age-associated disease characterized by the accumulation of senescent cells, particularly vascular smooth muscle cells (VSMCs), which remain metabolically active but exhibit significant morphological changes, mitochondrial dysfunction and adopt a senescence-associated secretory phenotype (SASP). Despite growing recognition of senescent VSMCs’ detrimental roles in atherosclerosis, like sustained local inflammation, matrix degradation, and tissue remodelling, their potential contribution to intraplaque angiogenesis through pro-angiogenic SASP has not yet been elucidated. Moreover, while statin therapy, the most widely used drugs for the prevention and treatment of atherosclerosis, plays a crucial role in managing atherosclerosis by reducing cholesterol levels and promoting plaque stability, its impact on mitochondrial dysfunction and SASP in senescent VSMCs remains largely unexplored. AIM: This study aimed to investigate the interplay between VSMC senescence, mitochondrial dysfunction, SASP, and angiogenesis, and to evaluate the effect of simvastatin on these processes, and to test whether targeting specific SASP-related pathways (VEGF/VEGFR2 and NF-κB) may represent more effective therapeutic strategies. RESULTS: We demonstrated, using both single-cell RNA sequencing datasets and immunohistochemistry, that senescent VSMCs are significantly more abundant in human unstable carotid atherosclerotic plaques than in stable ones. Consequently, based on these premises and in order to study senescent VSMCs more thoroughly, we developed and characterized two in vitro models of VSMC senescence: replicative (or old cells) and doxorubicin-induced mimicking stress-induced premature senescence, by assessing key senescence features such as cell morphology, cell cycle arrest, SASP, and mitochondrial dysfunction. Interestingly, both senescent models displayed an accumulation of dysfunctional mitochondria characterized by reduced mitochondrial membrane potential (MMP) and respiration, accumulation of reactive oxygen species (ROS), and an altered mitochondria morphology when compared to their young cells control. Moreover, TFAM and TOM70 expression was found to be downregulated only in old cells suggesting a reduction of mitochondrial biogenesis. In addition, both models of senescent VSMCs showed a significant increase in the gene expression and protein production of pro-inflammatory cytokines, including IL-1β, IL-6, and IL-8, thereby validating the SASP profile.Next, we investigated whether simvastatin could ameliorate age-associated phenotypes in senescent VSMCs. Simvastatin (0.1 μM) improved mitochondrial respiration and reduced ROS production and the inflammatory SASP in both doxorubicin-induced and old VSMCs.Notably, when mevalonic acid, a downstream product of the HMG-CoA reductase pathway, was added, these beneficial effects of simvastatin on mitochondrial function and the SASP were no longer observed. This indicates that simvastatin's impact on mitochondrial respiration and SASP is specifically mediated through the inhibition of HMG-CoA reductase, the enzyme targeted by statins in the cholesterol biosynthesis pathway.Multiplex analysis of the inflammatory SASP in the cell culture medium of the senescent VSMCs
THE IMPACT OF VASCULAR SMOOTH MUSCLE CELL SENESCENCE ON MITOCHONDRIA DYSFUNCTION AND ANGIOGENESIS / C. Rossi ; tutor: A. Corsini ; co-tutor: S. Bellosta ; coordinatore: G.D. Norata. - Milano. Dipartimento di Scienze Farmacologiche e Biomolecolari Rodolfo Paoletti, 2025 Dec 16. 38. ciclo, Anno Accademico 2025/2026.
CONCERTI PER TASTIERA DI SCUOLA NAPOLETANA E COMMITTENZA ARISTOCRATICA NEL SETTECENTO
This research aims at offering a musicological study ranging in its approach from historical, philological and critic perspectives, focusing on keyboard concertos of the Neapolitan school in the second half of the 18th century and presenting the edition of two previously unpublished works, composed by Ferdinando Pellegrini (1715 ca. – 1766 ca.) and Alessio Prati (1750-1788), found in Stockholm. Considering the 18th century concertos as expression of the civiltà della conversazione, these turn out to be crucial to the dialogue on which the instrumental language is based and the relevant connections with their dramaturgical aspects. Moreover, the musical system varies according to the existing relations inside the varying aristocratic patronage network – often promoted by high standing noble women, thus facilitating the approach towards gender history. Furthermore, essential relations with the freemasonry are not to be ignored nor excluded. Assuming a different point of view, these could be used to (re)interpret the tracks beaten by the artistic fate and path of some composers as part of their itinerancy.
CONCERTI PER TASTIERA DI SCUOLA NAPOLETANA E COMMITTENZA ARISTOCRATICA NEL SETTECENTO / A. Cantore ; tutor: C. Fertonani ; coordinatore: F. Slavazzi. Dipartimento di Beni Culturali e Ambientali, 2023 Jul 17. 35. ciclo, Anno Accademico 2022.
DEVELOPMENT OF NEW METHODOLOGIES AND PRACTICES TO REDUCE THE ENVIRONMENTAL IMPACT IN THE PRODUCTION OF CERAMIC MATERIALS
In response to the growing global challenges of energy security and water pollution, this thesis explores integrated electrochemical and photocatalytic approaches for sustainable hydrogen production and advanced wastewater treatment. By coupling the anodic oxidation of nitrogen- and pharmaceutical-based pollutants with cathodic hydrogen evolution, the research proposes a dual-purpose strategy that simultaneously enables pollutant removal and green energy generation.A key innovation of this work is the development and electrochemical evaluation of non-noble metal electrodes, particularly cobalt phosphide-based systems, which operate efficiently under alkaline conditions. These materials were tested in both ideal and contaminated environments, including real-matrix simulations containing diclofenac and Rhodamine B. The hydrogen production was directly quantified, providing, for the first time, a scientific assessment of the faradaic efficiency of hydrogen generation from real wastewater containing organic contaminants using non-precious electrodes.To overcome the limitations of electrochemical oxidation alone, a hybrid process integrating visible-light-active BiOCl photocatalysis was implemented, enhancing degradation kinetics and mineralization efficiency. This combined strategy offers a scalable and energy-efficient route for water remediation.Beyond laboratory scale, the study includes a case analysis of the industrial implementation of green hydrogen at Iris Ceramica Group’s H₂ Factory, where hydrogen produced via photovoltaic-powered alkaline electrolysis is blended with methane to fuel ceramic kilns. Energy balances, emission monitoring (via GC–MS), and product quality evaluations were conducted to assess the technical feasibility and environmental benefits of hydrogen integration in ceramic manufacturing.Overall, the thesis provides a multi-scale perspective—spanning materials design, process engineering, and industrial application—demonstrating how wastewater can serve as a valuable resource for both clean energy production and environmental protection. The findings contribute to advancing circular economy principles and accelerating the transition toward decarbonized, sustainable manufacturing.
DEVELOPMENT OF NEW METHODOLOGIES AND PRACTICES TO REDUCE THE ENVIRONMENTAL IMPACT IN THE PRODUCTION OF CERAMIC MATERIALS / E. Marcolini ; tutor: C.. L. M. Bianchi ; co-tutor: R. Zini ; phd coordinator: L. Prati. - Dipartimento di Chimica, via C.Golgi 19. Dipartimento di Chimica, 2025 Jul 17. 37. ciclo, Anno Accademico 2023/2024.
THE INFLUENCE OF MAOISM AND OF CHINESE COMMUNISM ON THE ITALIAN LEFT, 1949-1976
This work is about the influence of Maoism and of Chinese Communism on the Italian Left since 1949 until 1976.
THE INFLUENCE OF MAOISM AND OF CHINESE COMMUNISM ON THE ITALIAN LEFT, 1949-1976 / M. Gabbas ; tutor: D. Saresella. Università degli Studi di Milano, 2021 Mar 23. 33. ciclo, Anno Accademico 2020.
MOUNTAINHYDRO: INTEGRATING GEOLOGY, GEOPHYSICS AND HYDROGEOLOGY IN A MULTIDISCIPLINARY APPROACH
Spring water represents a vital source of drinking water for local communities in mountain areas worldwide. As water demand and pressure on groundwater resources increase in a context of changing climate, there is a growing commitment to assess groundwater availability and to develop sustainable water supply strategies.Effective groundwater management relies on a comprehensive characterization of aquifer systems. In this regard, karst-fractured terrains pose unique challenges due to their complex and heterogeneous nature. This study focuses on the assessment of aquifer systems in karst-fissured mountain environments, by employing a multidisciplinary approach based on three pillars: geology, geophysics, and hydrogeology.The geological pillar centers on the construction of a 3D geological model derived from a combination of existing geological maps and targeted field observations. The model captures the spatial distribution of geological units and major structural lineaments and serves as a framework for the interpretation of groundwater preferential flow paths. The geophysical pillar involves the use of high resolution DC resistivity surveys and densely spaced transient electromagnetic (EM) methods, both ground-based and airborne. Geophysical surveys play a key role in capturing the subsurface heterogeneity and identifying preferential flow paths within the rock volumes. The integration of the geological model into the geophysical inversion through an iterative process provides insights into the hydrogeological characteristics of the investigated system and enhances reliability of the interpretations. Moreover, the 3D inversion of airborne EM data has proven effective in increasing model accuracy in topographically and geologically complex terrains.The hydrogeological pillar consists of groundwater chemical and stable isotope analyses, as well as hydrodynamic characterization. These analyses support the delineation of recharge areas of the springs and help characterize the behavior of the aquifer system. In addition, numerical simulations of groundwater flow were carried out to test different scenarios of recharge and flow paths in order to validate the conceptual model.The results demonstrate the strength of an iterative, integrated modelling strategy in improving the understanding of complex karst-fissured aquifers.Two representative study areas were selected to implement and validate the multidisciplinary approach: one in northern Italy (Val Sabbia, Lombardy), and the other in southern Spain (Sierra de Aracena, Andalusia). These sites exemplify geologically complex mountain terrains, providing robust testing ground for the methodology.
MOUNTAINHYDRO: INTEGRATING GEOLOGY, GEOPHYSICS AND HYDROGEOLOGY IN A MULTIDISCIPLINARY APPROACH / A. Lucchelli ; advisor: D. Pedretti ; co-advisor: G. Fiandaca, R. Bersezio ; coordinator: G. Muttoni. Dipartimento di Scienze della Terra Ardito Desio, 2025 Jun 25. 37. ciclo
VALORIZZAZIONE DELL¿HERITAGE, CIRCOLARITÀ, VINTAGE E UPCYCLING.OPPORTUNITÀ PER LE AZIENDE PER UNA MODA PIÙ SOSTENIBILE
The contemporary fashion system, dominated by fast fashion and the characteristics of speed and obsolescence, seems to contrast with the concept of sustainability, today crucial for companies, institutions and consumers, all responsible actors for a more ethical and respectful future, from an environmental and social point of view. Although the challenges are complex, many companies are committed to developing sustainable products and to positively influence the system through innovation and dissemination.The research analyzes fashion as a complex system, which goes beyond production, and includes communication, consumer choices and alternative and circular options.The work examines the main critical issues of the sector to outline possible solutions. It studies cases of virtuous companies, with a focus on the Made in Italy that stand out for its ability to combine artisan tradition and innovation, enhancing the short supply chain and local skills. Significant examples include Ermenegildo Zegna, Rifò Lab and emerging brands that adopt innovative or upcycling strategies, proposing models in line with the needs of new generations. What emerges is the importance of a circular system that starts from design and ends with the management of end-of- life garments, for which companies and consumers are both responsible. Other relevant aspects that have emerged are: textile recovery, a historical Italian practice with great potential for the future; company archives, which through the conservation and reinterpretation of the past, offer inspiration for capsule collections, projects and a more conscious design, promoting quality and durability; the affirmation of second hand and vintage as increasingly popular options for consumers; the use of heritage in the strategies of some Italian luxury names, which demonstrates the central role of tradition for change; the importance of dissemination and training both towards the future generation of designers and towards consumers.A specific and in-depth study was conducted in collaboration with the OTB Group, focusing in particular on the Diesel brand, to analyze a brilliant example of a well-structured and diversified corporate sustainability strategy, which demonstrates how innovation and responsibility can guide the sector towards a more aware and responsible future.
"Valorizzazione dell’Heritage, Circolarità, Vintage e Upcycling.Opportunità per le aziende per una Moda più Sostenibile / S. Autorino ; tutor: E. Scarpellini ; coordinatrice: P. Grillo. - via santa sofia, milano. Dipartimento di Studi Storici ""Federico Chabod"", 2025 Jun 04. 37. ciclo, Anno Accademico 2023/2024."
BIOCHEMICAL INSIGHTS INTO THE PROTEIN NUMA AND ITS BINDING PARTNERS BETWEEN MITOTIC SPINDLE AND NUCLEAR COMPARTMENTS
During my PhD program I worked on two Nuclear Mitotic Apparatus (NuMA)-related projects. The first one centred on the biochemical and structural characterization of the NuMA-dynein mitotic interaction and was published last year on Structure. The second project focused on the study of the largely unknown role of NuMA in the nucleus during interphase. Regarding this part, I got interesting details on the NuMA-53BP1 (p53-binding protein 1) interaction in the context of liquid-liquid phase separation (LLPS).In multicellular organisms, the proper organization of the mitotic spindle is essential for accurate cell division, tissue development and homeostasis. In vertebrate cells, the protein NuMA is a master regulator of mitotic spindle functions, implicated in spindle assembly and orientation, working together with the high molecular weight dynein-dynactin microtubule-motor complex. The domain structure of NuMA consists of an N-terminal globular domain, a central extended coiled-coil, and an unstructured C-terminal cargo-binding region. Whether NuMA is a dynein-dynactin activating adaptor is still not known. On these premises, the first part of my PhD project focused on the characterization of the NuMA-dynein binding interface, which I performed in collaboration with other members of the group. The crystal structure of the N-terminal head of NuMA (NuMA_1-153) revealed that it folds into a hook domain, a conserved feature of the Hook-family dynein-dynactin adaptors interacting directly with the Light Intermediate Chain (LIC) subunit of dynein. Pulldown assays performed with purified proteins indicated a direct interaction between NuMA_1-705 and LIC and identified four conserved residues in the NuMA hook domain that are crucial for LIC binding. Interestingly, sequence alignment between NuMA and known CC1-box containing dynein-dynactin adaptors revealed the existence of a CC1-box-like motif in the NuMA N-terminal coiled-coil domain (NuMA_365-376) that we demonstrated to be also implicated in contacting LIC. Thus, our studies identified two sites on NuMA’ N-terminus required for the interaction with a conserved hydrophobic helix in LIC1 C-terminus. Spindle positioning assays in human HeLa cells showed that these newly identified dynein-binding interfaces of NuMA are essential for correct mitotic progression. Collectively, these results support the notion that NuMA acts as a mitotic dynein-dynactin adaptor, forming complexes with similar topology to what observed for other known hook and CC1-box containing adaptors.In vertebrate cells, NuMA accumulates in the nucleus during interphase and contributes to the DNA damage response (DDR), negatively regulating the 53BP1 double strand break (DSB) repair function. The second part of my PhD project focused on the characterization of the NuMA-53BP1 binding interface. By co-immunoprecipitation (co-IP) experiments in human HEK293T nuclear extracts with anti-NuMA antibodies, I confirmed that endogenous NuMA interacts with 53BP1, and that this interaction is decreased upon DNA damage induction. Interestingly, analytical size-exclusion chromatography (SEC) experiments with purified fragments revealed that the C-terminus of 53BP1 (53BP1_1484-1972) interacts directly with the C-terminus of NuMA (NuMA_1821-2115). These are two intrinsically disordered domains, common to proteins that undergo LLPS, a mechanism conferring spatial and temporal regulation to biological processes. Since 53BP1 forms DNA damage foci, which are LLPS condensates promoted by its C-terminal disordered region, I tested whether also NuMA is involved in this mechanism. Interestingly, I found that NuMA_1821-2115 forms liquid droplets in vitro at 20 uM and physiological salt concentrations, promoted by electrostatic and polar interactions. By co-IP experiments in HEK293T nuclear extracts, I also detected an interaction of NuMA with the MT nucleator TPX2. Since TPX2 counteracts the 5
BIOCHEMICAL INSIGHTS INTO THE PROTEIN NUMA AND ITS BINDING PARTNERS BETWEEN MITOTIC SPINDLE AND NUCLEAR COMPARTMENTS / C. Renna ; supervisor: M. Mapelli ; internal advisor: T. Bonaldi ; phd coordinator: S. Minucci. Università degli Studi di Milano, 2021 Dec 13. 33. ciclo, Anno Accademico 2021.
POTENTIAL GENE VARIANTS INFLUENCING FVIII LEVELS: THE ROLE OF LDL RECEPTOR ON FVIII CLEARANCE.
BackgroundGenetic variants in the LDL receptor (LDLR) gene have been associated with higher LDL cholesterol levels, premature atherosclerotic cardiovascular disease (ASCVD) and increased cardiovascular risk. However, the increased arterial thrombotic risk observed in LDLR carriers is partly independent from the lipoprotein levels and could be related to the LDLR modulation of FVIII levels. Indeed, a role of LDLR in FVIII clearance has been shown and LDLR variants have been associated with higher FVIII levels.To date, it is not clear which are the LDLR variants involved in FVIII clearance and the underlying mechanism. With this background and gap of knowledge, the present study aims to: - identify the LDLR variants associated with high FVIII coagulant activity (FVIII:C≥150 IU/dL) in a cohort of DVT (Deep Veen Thrombosis) patients and healthy control subjects; - functionally and biochemically characterize the LDLR variants (identified in this study and described in literature) associated with high FVIII levels in order to understand their ability to modulate the FVIII clearance, in terms of FVIII binding and/or uptake.MethodsWe selected 596 Italian subjects (298 DVT patients and 298 controls) enrolled and sequenced in the frame of the DVT-Milan study. FVIII:C was measured by one-stage clotting assay. A multiplexed next generation sequencing was performed. In order to select the variants to be characterized in vitro, we analyzed the association between LDLR variants and high FVIII:C (FVIII:C≥150 IU/dL). For common and low-frequency LDLR variants a logistic regression model was performed. Rare LDLR variants have been analyzed using cumulative association tests.Wild type (WT) and selected mutant LDLR vectors were transiently transfected in LDLR-deficient Chinese hamster ovary (CHOldlA7) cells with chemical methods. The LDLR expression in total cell lysates was assessed by western blotting (WB). Immunofluorescence (IF) was performed to evaluate total, surface and intracellular expression of LDLR proteins.ResultsIn our cohort of DVT patients and healthy control subjects we identified the missense LDLR variant rs45508991 associated with high FVIII levels (OR=5.47, p=0.06). We selected 4 LDLR variants to be characterized in vitro: the variant rs45508991 and 3 synonymous variants (rs688, rs2228671 and rs5925), previously reported to be associated with higher FVIII levels or increased risk of ischemic stroke and/or coronary artery disease. Among the 4 LDLR variants expressed in vitro, we observed, by both WB and IF, a significant reduced expression of total LDLR protein carrying the rs45508991 variant, compared to the WT LDLR. Moreover, by IF, we observed that the surface LDLR expression was significantly reduced in cells expressing the variant rs45508991.Conclusions and perspectivesOur hypothesis is that a reduced expression of LDLR, total and on cell surface, caused by the identified variant rs45508991, could impair FVIII binding and clearance, leading to an increase in FVIII levels. In order to test this hypothesis, we will perform the fluorescence-activated cell sorting (FACS) with the goal to confirm IF findings concerning the cell surface LDLR expression in all samples and to evaluate the LDLR-mediated binding of FVIII.
POTENTIAL GENE VARIANTS INFLUENCING FVIII LEVELS: THE ROLE OF LDL RECEPTOR ON FVIII CLEARANCE / P. Agosti ; tutor: F. Payvandi ; supervisori: S. Spena, A. Cairo ; coordinatore: C. Sforza. Dipartimento di Fisiopatologia Medico-Chirurgica e dei Trapianti, 2022 Jun 07. 34. ciclo, Anno Accademico 2021.
TOWARDS ACCELERATION OF QUANTUM-CLASSICAL ORCHESTRATION
Quantum computing is expected to act as a hardware accelerator within future computing infrastructures, complementing classical processors in the same way GPUs accelerate specific workloads today. While algorithms such as Shor’s and Grover’s suggest potential advantages, the practical use of quantum devices will occur within hybrid architectures where classical and quantum resources coexist. Artificial intelligence illustrates this balance: classical methods, such as Transformers and their scalable variants, remain dominant, while quantum computing may only contribute to selected subroutines. At the same time, AI methods are increasingly applied to quantum computing, supporting calibration, control, and error mitigation of near-term devices. This two-way relationship underlines the need for hybrid solutions and for efficient orchestration of heterogeneous resources. This thesis investigates the design and implementation of hybrid classical-quantum solutions, with emphasis on their real-time orchestration within heterogeneous infrastructures. The work has been carried out within the Qibo environment, a fully open-source framework that spans the entire quantum computing stack: from high-level interfaces for algorithm design, to efficient simulation backends on classical hardware, to integration with quantum devices down to the hardware and pulse level on self-hosted systems. This modularity makes Qibo a suitable platform to explore orchestration strategies across classical and quantum resources.Within this framework, we introduce algorithmic and software contributions that accelerate hybrid workloads. We present general-purpose open-source libraries, such as Qiboml for hybrid quantum-classical machine learning, and mpstab for a hybrid stabilizer-tensor network representation of quantum states. By preserving the familiar interfaces of classical machine learning frameworks, Qiboml enables the design of hybrid architectures and their execution on both simulators and quantum hardware. We also develop algorithmic solutions for real-time quantum error mitigation, implemented in Qiboml and relying on Qibo. We further discuss applications, including multi-variable integration, probability density estimation, and ground-state search, as well as an application in which quantum systems operate as precision sensors for fundamental physics. These serve as test cases to validate the proposed approaches and to provide practical examples of how the heterogeneous components of a future quantum-classical infrastructure can be effectively orchestrated.
TOWARDS ACCELERATION OF QUANTUM-CLASSICAL ORCHESTRATION / M. Robbiati ; supervisore: S. Carrazza, S. Vallecorsa ; coordinatore: A. Mennella. Dipartimento di Fisica Aldo Pontremoli, 2025 Nov 28. 38. ciclo, Anno Accademico 2024/2025.
MOLECULAR DETERMINANTS UNDERLYING PROTEIN MISFOLDING AND AGGREGATION
Proteins have evolved to adopt distinctive and well-defined functional states under physiological conditions, either as monomers or complexes. The achievement of a three-dimensional structure allows proteins to exert their physiological functions. Nevertheless, when proteins lose – or fail to acquire – their spatial organization, they can convert into aggregated species that can be harmful to the organism. Conformational diseases gather all those pathologies characterized by the misfolding and aggregation of proteins. Indeed, while the formation and deposition of proteinaceous aggregates can be toxic to cells, the lack of active folded protein disrupts normal physiological pathways. Although considerable progresses have been made in the recent years, to date conformational diseases are still incurable. Indeed, the incomplete understanding of the causes guiding protein misfolding and aggregation prevents the development of efficient treatments. At the same time, the complexity and the diversity of the processes leading to the formation of aggregated species make the task extremely challenging.This PhD project was developed to provide a more comprehensive overview of the molecular bases underlying the conversion of soluble and functional states into aggregated and potentially toxic species. To reach such aims, we applied an integrative approach on two model systems, neuroserpin (NS) and beta-2 microglobulin (2m). In particular, we combined a series of biophysical, biochemical and structural techniques to study these two proteins which have been largely used as model systems for serpin polymerization and amyloid formation, respectively. We found that protein misfolding and aggregation processes depend on several molecular properties, including primary sequence, denatured state compactness, thermal stability, ability to form oligomers under physiological conditions, and the presence of post-translation modifications.The data presented in this PhD thesis add valuable information to depict the complex framework of protein misfolding and aggregation.
MOLECULAR DETERMINANTS UNDERLYING PROTEIN MISFOLDING AND AGGREGATION / L. Broggini ; scientific tutor: S. Ricagno. Dipartimento di Bioscienze, 2021 Apr 22. 33. ciclo, Anno Accademico 2020. [10.13130/broggini-luca_phd2021-04-22].
MODIFYING PHYLLOTAXIS IN BRASSICA SEED CROP SPECIES FOR YIELD IMPROVEMENT
In a world that is continuously growing and developing, it is crucial to enhance agriculturalyields to ensure an adequate food supply for the population while minimizing environmentalimpact. Plant yield is influenced by a range of factors, among which inflorescencearchitecture and its geometrical organization, known as phyllotaxis. Despite centuries ofstudies, the mechanism behind phyllotactic patterning is still partially unknown, especiallyin non-model plant species. Furthermore, the direct impact of phyllotaxis on overall yieldis not well understood.In this thesis, we started to tackle these knowledge gaps. First, we reported support to thehypothesis of a direct link between phyllotaxis and yield by describing several ArabidopsisREM mutants with a permutated phyllotaxis and an increased number of siliques on the mainstem.Consequently to this result, we aimed to investigate the conservation of such a mechanismin the economically important crop Brassica napus.We identified the candidate homologous genes in rapeseed and confirmed their expressionin reproductive tissues. Moreover, we demonstrated that they can influence phyllotaxis inArabidopsis, suggesting a retained role between these two species. Protein interaction testsfurther suggested that some interaction between REMs themselves and the auxin pathwaycan be conserved.To finally prove the mechanism in rapeseed, we aimed to generate the homolog REM mutantsin the species by CRISPR-Cas9 gene editing. After an in silico and experimental selection ofthe gRNAs, we focused on the Agrobacterium mediated transformation of rapeseed.According to our results, both the transformation protocol as well as the editing need to beoptimized if we want to apply up to date molecular techniques in Brassica napus.Indeed, we detected inefficiency and high technical requirements in the transformationprotocol and a low functionality of the editing machinery in transformants, the latter beingreported in literature too.Optimizing such techniques can exponentially speed up basic and applied research inrapeseed and open new possibilities in crop design. Indeed, multiple mutants are very oftenrequired to get phenotypes in high redundant genomes such as the one of Brassica napusand producing them by random mutagenesis is highly time consuming.Overall, this application-driven study unveils novel genes associated with phyllotaxis andyield in the Brassicaceae family, contributing to our understanding of plant architecture andoffering insights into sustainable strategies for improving crop productivity.
MODIFYING PHYLLOTAXIS IN BRASSICA SEED CROP SPECIES FOR YIELD IMPROVEMENT / C.c. Ferrario ; scientific supervisor: M. M. Kater ;tutor: V. Gregis, F. Caselli ; PhD school coordinator: R. Mantovani. - dipartimento di bioscienze - Università degli studi di Milano. Dipartimento di Bioscienze, 2025 Mar 31. 37. ciclo, Anno Accademico 2023/2024.
THE HISTONE POST-TRANSLATIONAL MODIFICATION LANDSCAPE IN HPV+ AND HPV- HEAD AND NECK SQUAMOUS CELL CARCINOMA: CHARACTERIZING THE ONCOGENIC ROLE OF THE H3K36ME2 METHYLTRANSFERASE NSD2
Background: HNSCC is a heterogeneous group of tumors caused mainly by environmental factors and human papillomavirus (HPV) infections. HPV- and HPV+ HNSCC are considered distinct entities, however, they are still treated with the same therapeutic strategies. HPV-induced tumorigenesis is mainly mediated by the E6/E7 oncoviral proteins, that, among all, alter the epigenetics of the host cells. Nevertheless, epigenetic profiles of HNSCC subtypes have not been clearly profiled. Results and Conclusion: hPTMs super-SILAC analysis of HNSCC cell lines and patients’ tissue samples revealed significant differences in the enrichment levels of some hPTM in HPV+ samples compared to HPV- ones and in tumoral tissues compared to normal ones. We focused on one of these identified hPTM and demonstrated that its levels are regulated by E6 and E7. We identified a histone modifier responsible for this hPTM whose levels are upregulated by E6/E7 and are higher in HPV+ compared to HPV- HNSCC cell lines and patients’ tissue samples, as is for the related hPTM. Silencing this enzyme through shRNA in HNSCC cell lines reduced proliferation and migration rates in both subtypes. It also downregulates the expression levels of some EMT mesenchymal makers and of a crucial oncogene involved in HNSCC. RNA-seq analysis revealed that other programs are instead specifically regulated according to the subtype: immune-response related genes are mainly activated in HPV- cell lines, while genes involved in cell differentiation in the HPV+ ones. Our research paves the way to novel lines of research and identifies a promising novel epigenetic target for HNSCC treatments.
THE HISTONE POST-TRANSLATIONAL MODIFICATION LANDSCAPE IN HPV+ AND HPV- HEAD AND NECK SQUAMOUS CELL CARCINOMA: CHARACTERIZING THE ONCOGENIC ROLE OF THE H3K36ME2 METHYLTRANSFERASE NSD2 / L. Ghiani ; tutor: S. Chiocca ; co-tutor: M.Alcalay ; internal advisor: S. Minucci ; external advisor: G.P. Dotto ; phd coordinator: G. Viale. Dipartimento di Oncologia ed Emato-Oncologia, 2021 Mar 30. 32. ciclo, Anno Accademico 2020.
FUTURE PERSPECTIVES IN NON-ADVANCED AGE-RELATED MACULAR DEGENERATION: FROM MULTIMODAL IMAGING APPROACH TO ESTABLISHING CORE OUTCOME MEASURES
Age-related macular degeneration (AMD) is the leading cause of irreversible vision loss in developed countries, and its prevalence is expected to rise due to increasing life expectancy. This doctoral dissertation focuses on the earlier stage of the disease, known as non-advanced AMD, during which vision is largely preserved. There is growing evidence that this stage should be targeted for early treatment to manage this chronic condition effectively.Following a comprehensive literature review of non-advanced AMD, the first study explores the role of multimodal imaging in the early detection of retinal aging, using a novel, non-invasive technique called retromode imaging. The findings indicate that retromode imaging detects significantly more drusen-like deposits (DLDs) than conventional fundus photography in the maculae of healthy individuals. Importantly, these DLDs were not associated with pathological changes, as their frequency increased with normal aging.The second study assesses the accuracy and reproducibility of imaging modalities in delineating reticular pseudodrusen (RPD) areas. The results identify the combined use of structural optical coherence tomography (OCT) and near-infrared reflectance (NIR) as the most reliable method for RPD detection.The third study investigates the relationship between macular pigment optical volume (MPOV) and retinal layer thickness, suggesting that outer nuclear layer (ONL) thickness could serve as a biomarker for macular pigment levels. However, the need for further research into the effects of AMD on MPOV is emphasized.Finally, a Delphi consensus process was employed to establish a minimum set of outcome measures for non-advanced AMD, paving the way for the development of an international, web-based registry. This registry will enable the collection of real-world data to enhance understanding of the disease’s natural progression and facilitate the evaluation of potential therapeutic interventions.Overall, the findings presented in this dissertation provide a foundation for future research and clinical innovations aimed at earlier intervention and improved patient outcomes.
FUTURE PERSPECTIVES IN NON-ADVANCED AGE-RELATED MACULAR DEGENERATION: FROM MULTIMODAL IMAGING APPROACH TO ESTABLISHING CORE OUTCOME MEASURES / M. Cozzi ; tutor: G. Staurenghi, A. Invernizzitutor esterno: S. Zweifelcoordinator: F. Cheli. Dipartimento di Scienze Biomediche e Cliniche, 2024 Jan 13. 37. ciclo, Anno Accademico 2023/2024.
INNOVATIVE IMAGING OF URINARY SYSTEM IN CANINE AND FELINE PATIENTS
Diseases of the urinary system are regularly encountered in daily veterinary practice. The development of increasingly efficient diagnostic tools is crucial to meet the high-quality requirements of contemporary professional standards.This project had many purposes, aiming to describe pioneering methods, protocols and diagnosis related to imaging of ureters and urinary bladder, chosen as they represent daily diagnostic challenges in daily routine practice.This project consisted of three papers: the first paper is a prospective pilot project concerning quantitative CEUS exam applied to distinguish neoplastic and non-neoplastic lesions of the urinary bladder in small animals; the second paper is a multicentric retrospective observational project describing the CT appearance of a novel CVC congenital malformation; the third paper is a retrospective study conducted on canine healthy patients, aimed at assessing the visibility of the ureters on high field MR on T1 and T2 sequences avoiding the use of paramagnetic contrast agents.The results of this project allowed to obtain objectifiable parameters for the distinction of neoplastic and non-neoplastic lesions of urinary bladder using quantitative CEUS; we also described the CT appearance of the transcaval ureter, a malfomaration of the CVC never described in veterinary medicine; finally, we described the feasibility of evaluation of normal ureters through high-field MR on T2-weighted sequences, in healthy canine patients.In conclusion, this project allowed to describe new diseases that could affect urinary tract function and contributes to the development of new methods and protocols with the potential to reduce the invasiveness of certain diagnostic procedures related to the urinary tract.
INNOVATIVE IMAGING OF URINARY SYSTEM IN CANINE AND FELINE PATIENTS / C. Spediacci ; revisori interni: R. FERRARI, E. ZUCCA, P. CAGNARDI ; revisori esterni: G. GNUDI, S. CITI ; supervisor: M. DI GIANCAMILLO ; co-supervisor: M. LONGO. Dipartimento di Medicina Veterinaria e Scienze Animali, 2023 Feb 07. 35. ciclo, Anno Accademico 2022.
ORGANOIDS AS TOOLS TO STUDY BRAIN DEVELOPMENT AND DISEASE
Human brain organoids (ORG) are a widely used in vitro model that contribute to our knowledge about the biology and pathophysiology of the nervous central system (CNS). They are 3D multicellular clusters that mimic the cytoarchitecture and the developmental pathways that occur in vivo. The most debated limitation of ORG is the lack of the endothelial component. Thus, at first we have developed a co-culture in vitro system composed by the Blood Brain Barrier (BBB) and ORG. The in vitro BBB is composed by human brain microvascular endothelial cells (HBMEC) and human astrocytes (HA). ORG are generated from induced pluripotent stem cells (iPSCs) cultured with different differentiation media for 36 days, and then co-cultured with the in vitro generated BBB.To evaluate the strength of our model, organoids in the absence (ORG) or in the presence (BBB-ORG) of the BBB were compared for their maturation stage and BDNF levels. In addition, the effects of magnesium (Mg) were investigated with a focus on excitotoxicity and neurodevelopment. Indeed, Mg is known to play crucial role in the CNS. It protects the CNS from inflammation, from excitotoxicity driven by NMDA-R over-activation, it has positive effects on the BBB integrity and it is involved in brain development. However, the mechanisms of action are still poorly understood and studied.To shed light on the Magnesium (Mg)-driven mechanisms involved in brain development and protection, an inorganic and an organic Mg salt (Mg sulphate, MgSO4, and Mg pidolate, MgPid, respectively) were added to the culture media to reach the extracellular concentrations of 1, the physiological extracellular Mg concentration, or 5 mM, to mimic Mg supplementation. After the treatment, the possible modulation of molecules involved in excitotoxicity and neurodevelopment was investigated by qRT-PCR, western blot or ELISA. In particular, we analysed the modulation of NMDA-R, GABA-R and their neurotransmitters glutamate and GABA, neurotrophic factors, dopamine and its receptors and the Mg channels TRPM6 and TRPM7. The maturation stage and the cortical layer organization were assessed by immunofluorescence using CTIP2, TBR2 and SOX2 antibodies to detect the cortical, the subcortical neurons and the neural progenitor cells, respectively. Toluidine blue staining and ultrastructural analysis using transmission electron microscopy were performed to further confirm the results.6Our data demonstrate that BDNF levels are higher in the BBB-ORG model because of the presence of the endothelial component, which is the main responsible for its secretion. Moreover, the cortical layer is more organized in the presence of the BBB than in its absence, underlying its importance.High concentrations of Mg salts lowers the levels of NMDA-R and increases GABA-Rs levels only in the presence of the BBB, thus suggesting a role in Mg in counteracting excitotoxicity. These results suggest that Mg supplementation might ameliorates the outcome of patients when administered as adjunctive therapy. Particularly, MgPid performs better than MgSO4. Interestingly, high concentration of Mg salts concentration results in a higher maturation stage of the BBB-ORG. The increased levels of released and intra-org BDNF detected in the presence of 5 mM MgPid could explain the faster maturation of the ORG.This study confirms that the BBB-ORG is a useful tool to study the crosstalk between BBB and brain in both physiological and pathological conditions, and emphasises the importance of the continuous search for a model that closely resembles the in vivo context. Moreover, although further analyses are needed, these studies unveil the role of Mg in brain development and in modulating the release of BDNF from the BBB.In addition, preliminary studies on the neuroinvasiveness of SARS-Cov-2 were conducted on ORG compared to BBB-ORG, and also on HBMEC and HA alone. Our results reveal once again that the presence of the BBB is fundamental for in vitro
ORGANOIDS AS TOOLS TO STUDY BRAIN DEVELOPMENT AND DISEASE / G. Fedele ; coordinatore di dottorato: N. Landsberger ; tutor: S. Castiglioni ; external advisor: F. Rios. Dipartimento di Scienze Biomediche e Cliniche, 2025 Jan 21. 37. ciclo, Anno Accademico 2023/2024.
VALORISATION OF BIOMASS-DERIVED MOLECULES BY NOBLE METAL CATALYSTS
Valorisation of different biomass derived molecules was successfully approached and studied in this PhD project. The focus of the thesis was addressed to the catalysts preparation, passing through an accurate catalytic designed, to be then tested in academic and industrially appealing reactions. This approach led to the synthesis of different but equally interesting catalytic systems for the valorisation of substrates derived from the first and second generation of biomass feedstock.An extended study, at first, was conducted on the oxidation of glycerol (1st generation of biomass related), both in alkaline (needed for gold monometallic systems) and free pH (high industrial relevance) conditions. The target reaction was approached starting from the simplest Au/C catalytic systems, to finally move to more complicated and innovative materials: bimetallic once. Initially, the Au on carbon Vulcan (with the highest graphitisation degree) SOL derived catalysts showed a remarkable initial activity (IA= 1091 h-1) in comparison with the other carbonaceous supports (Norit and X40S) and the SMAD derived catalysts. This result pointed out the importance of the protecting agent (a polymer that surrounded the nanoparticles and is solely present for the SOL synthetic route) beside the importance of the support’s features. Similarly, electronic effects ascribed to the interaction with the support of the nanoparticles (i.e. the strong metal support interaction (SMSI) thermally induced on Au4Ag1/TiO2) showed to be the ruling factor to determine the oxidation state of the metals. This latter, subsequentially, influence the catalytic activity: an enhanced initial catalytic activity was detected for the Au4Ag1/TiO2 catalyst (IA= 1616 h-1), in comparison with the Au4Ag1/Al2O3 (IA= 963 h-1). The SMSI have influenced also the stability of the system, avoiding the enlargement of the nanoparticles during the thermal treatments. On the other hand, the SMSI induced the presence of Ag+ species onto the bimetallic nanoparticles titania supported, leading to a quite rapid deactivation of the catalytic system. The thermal treatments pointed out also the importance of the protecting agent (polyvinyl alcohol, PVA): on one side when it is present confers resistance to the system towards the nanoparticles aggregation, on the other when it is removed from the nanoparticles’ surface (by the same thermal treatment), the catalyst acquired an enhanced initial activity. AuPt/TiO2 catalytic systems were subsequentially exploited both in alkaline and free pH conditions. The gold content positively influenced the activity of the catalytic systems in both the conditions. In particular Au9Pt1/TiO2 was the most active catalyst in the alkaline condition (IA= 7389 h−1), and Au6Pt4/TiO2 showed the highest initial activity (IA= 301 h-1) in free pH condition. For all the bimetallic system mentioned and exploited in the valorisation of glycerol, furthermore, a synergistic effect was detected. The importance of gold as modifier to confer resistance to the catalytic system by stabilizing the oxidation state of the second metal was also established.Subsequentially, completely different designed and synthesised catalysts were prepared for the valorisation of substrates related with the 2nd generation of biomass. Bare carbon nanofibers (CNFs) and functionalised CNFs (CNFs-P, CNFs-O and CNFs-N), for instance, were employed as supports for Ru nanoparticles (introduced by incipient wet impregnation). All the catalysts prepared showed activity in the valorisation of cellulose derived molecules. In particular, it was observed how N-containing functionalisation of the support, promoted by a strong interaction with the Ru nanoparticles, led to the highest catalytic activity among the set of catalysts tested for the levulinic acid (LA) hydrogenation (88 % of conversion after 3 h) with a full selectivity to y-valerolactone (GVL). On the
VALORISATION OF BIOMASS-DERIVED MOLECULES BY NOBLE METAL CATALYSTS / A. Jouve ; Tutor: L. Prati ; Co-tutor: C. L. Bianchi ; coordinatore: M. Pizzotti. Università degli Studi di Milano, 2020 Jan 28. 32. ciclo, Anno Accademico 2019. [10.13130/jouve-andrea_phd2020-01-28].
MEASURING AND MODELLING COVER CROP GROWTH AND AGRONOMIC EFFECTS
Cover crops are cultivated during the bare soil period between the harvest of a cash crop and the sowing of the next one. Their cultivation puts into effect the permanent soil organic cover principle of conservation agriculture and exerts several agro-ecological services, among which the most relevant are nitrate leaching reduction, weed growth control, soil organic matter increase, soil structure and water infiltration improvement. In temperate climates when crop rotations include summer cash crops (such as maize or soybean), autumn-winter cover crops are sown between late July and October and terminated from March to April of the following year. When sown in autumn, frost-sensitive cover crops may also be terminated efficiently by frost damage: this termination method is frequently called ‘winterkill’. Black oat (Avena strigosa Schreb.) and white mustard (Sinapis alba L.) are two of the most interesting and widespread frost-sensitive cover crops due to their adaptability to various environmental conditions and cropping systems. Even if these species are widely adopted as cover crops, there is a lack of information concerning both crop management and agronomic effects, as well as winterkill termination occurrence frequency and efficiency in temperate climates. Dynamic cropping systems simulation models can be used to determine crop management scenarios convenience for a wide range of weather and soil conditions, while the field trial assessments require large resource investments. However, the application of a simulation model to white mustard and black oat cover crops presents several knowledge gaps, as the limited number of studies focused on winterkilled cover crops growth and agronomic effects carried out in northern Italy. Furthermore, an integrated simulation model dealing both with cover crop growth and winterkill termination, and its consequent effect on the crop-soil system, including cover crop residue degradation on soil surface, is lacking. This work aimed at representing, within the ARMOSA cropping system model framework, frost sensitive cover crop species growth, development, and agronomic effects by enriching the simulation model with two additional modules, dealing respectively with winterkill events and cover crop superficial residue decomposition. The model was calibrated for white mustard and black oat cover crops using experimental data deriving both from a three-year field trial, from a commercial field monitoring campaign and from previous experiments, carried out in the region of interest. During the three-year field trial, white mustard, black oat and their mixture with purple vetch (Vicia benghalensis L.) have demonstrated a good aboveground biomass production potential (2-3 t DM ha-1), particularly when planted before the first half of September. Their nitrogen uptake (45 kg N ha-1 on average, up to 148 kg N ha-1) follows the biomass accumulation patterns, while their weed species control ability has proven to be consistently high. Overall, the improved ARMOSA model correctly simulated these species development (RRMSE equal to 27.3 and 29.5% respectively for black oat and white mustard), as well as soil water content and temperature (RRMSE equal to 8.4% and 19.2%). The employment of the new ARMOSA version to simulate black oat and white mustard cultivation, generally improved significantly both aboveground biomass simulation (RRMSE was decreased by 56.3% in comparison to the use of the original model version), leaf area index (RRMSE reduction of 31.6%) and C:N ratio simulations (RRMSE reduction of 8.8%). The convenience of the new model version employment was assessed in a wide range of sites (six sites of several provinces of Lombardy region in northern Italy), pedological conditions (soil textures from sandy-loam to silty-clay), weather conditions (calibration seasons ranged from 2019/2020 to 2021/2022) and management practices (minimum and no til
MEASURING AND MODELLING COVER CROP GROWTH AND AGRONOMIC EFFECTS / M. Gabbrielli ; tutor: L. Bechini; coordinatore: P.A. Bianco. Dipartimento di Scienze Agrarie e Ambientali - Produzione, Territorio, Agroenergia, 2022 Dec 09. 35. ciclo, Anno Accademico 2022.
Shape coexistence in even-even Sn nuclei with mass number A=110-120
Shape coexistence is a ubiquitous phenomenon across the nuclear chart, characterized by the appearance of states with different intrinsic shapes at similar excitation energies. In tin isotopes (Z=50), theoretical models predict such coexisting excitations, and experimental evidence for intruder configurations was first observed in the 70s. Nevertheless, spectroscopic information on low-lying 0⁺ excited states, essential to shed light on the onset of shape coexistence in this region, has remained limited.This PhD thesis presents a systematic study of low-lying 0⁺ excitations in even-even 110-120Sn isotopes, combining γ-ray spectroscopy with lifetime measurements. A series of experiments was performed at the Horia Hulubei National Institute for R&D in Physics and Nuclear Engineering (IFIN-HH) with the ROSPHERE-SORCERER setup and at the Legnaro National Laboratory (LNL) with the AGATA-PRISMA setup, employing heavy-ion transfer reactions to populate excited states. Particle-γ coincidence techniques enabled detailed spectroscopic studies, while lifetime measurements were performed using the Doppler Shift Attenuation Method and the Recoil Distance Doppler Shift method.The deduced transition probabilities significantly expand the experimental knowledge of 0⁺ states in this mass region, revealing systematic trends that support strong configuration mixing and the presence of coexisting shapes. These findings provide important constraints for modern nuclear-structure models and contribute to a deeper understanding of shape coexistence in medium-heavy nuclei.
Shape coexistence in even-even Sn nuclei with mass number A=110-120 / G. Corbari ; supervisor: S. Leoni ; co-supervisor: A. Bracco ;coordinatore: A. Mennella. - Dipartimento di Fisica, Università degli Studi di Milano. Dipartimento di Fisica Aldo Pontremoli, 2025 Dec. 38. ciclo, Anno Accademico 2024/2025.
IL CONTENZIOSO EUROPEO IN MATERIA DI AIUTI DI STATO FRA ESIGENZE DI TUTELA DELLE IMPRESE E DISCREZIONALITÀ DELLA COMMISSIONE
Ever since the Spaak Report, State aid control, considered to be one of the key policies for the creation of the common market, has relied on an architecture aimed at a continuous and rigorous monitoring of public concessions, with the ultimate goal of avoiding State behaviour detrimental to the internal market. To this end, the Treaties entrust to the (exclusive) competence of the European Commission the assessment, on a case-by-case basis, of the compatibility of the state measure with the internal market, preventing it from being implemented – and, therefore, from producing potentially distorting effects – before its compatibility with the Union framework has been verified. The Commission thus enjoys a wide discretionary power both in assessing the authorisation of support measures adopted by Member States and in relation to the enforcement of the discipline, being able to authorise exceptions to the general prohibition of State aid, weighing national objectives against those of European integration. The importance of these two actors tends, however, to obscure the significant impact that the application of State aid rules has on private undertakings, whose interests risk, however, being significantly compromised. In particular, as regards the aid beneficiary, although it is often the actor that faces the highest financial risk, it is not always in a more favourable position than other third parties involved in the control procedure. Against this background, the aim of this research is to examine, mainly through an in-depth examination of the Commission’s practice and the Court of Justice’s case law, the protections offered to (aid recipient) undertakings involved in the State aid control system by the European legal framework, attempting to delineate the quantum of protection provided to them by EU law. The aim of the work is to explore the extent to which beneficiary undertakings can rely on the State aid rules to protect themselves from the potentially harmful effects of Commission measures and whether – and ifso, how – the wide discretion enjoyed by the Commission affects the position of such undertakings to a greater or lesser extent.The analysis will concentrate, specifically, upon some critical aspects relating, on the one hand, to the protection accorded to the parties involved in the administrative procedure of State aid control before the European Commission; on the other hand, to the judicial protection offered in litigation before the Court, delving into beneficiaries’ access to appeals against the Commission’s decisions and the judicial review exercised by the Court on these decisions.
IL CONTENZIOSO EUROPEO IN MATERIA DI AIUTI DI STATO FRA ESIGENZE DI TUTELA DELLE IMPRESE E DISCREZIONALITÀ DELLA COMMISSIONE / F. Cagossi ; tutor: D. Diverio ; coordinatore: F. Biondi. Dipartimento di Studi Internazionali, Giuridici e Storico - Politici, 2024 Dec 18. 37. ciclo, Anno Accademico 2024.
DEMOCRACY AND JUSTICE IN COLLECTIVE ACTION INITIATIVES IN THE ENERGY FIELD
With the deployment of renewable energy technologies, the energy market is going towards a polycentric system characterized by the cohabitation of large- and small-scale installations. This configuration allows for the presence of new actors, fostering the deployment of renewable energy and representing a new tool to fight against climate change. This thesis investigates the role of one of these new players: energy communities. Energy communities represent a form of collective management based on the principle of “one people, one voice”, where citizens are the direct owners of energy, contrasting thus with the previous energetic model based on fossil-fuel energy and big companies and states. Furthermore, energy communities, intensely caring about social issues and inclusivity within their initiatives, imply political and social shifts. In this, they are related to energy democracy and justice, allowing all citizens to participate directly in the energy transition. This research inquiry analyses the reality of these initiatives, investigating whether possible gaps emerge between the citizen and the energy communities’ characteristics for potential inclusiveness at a time when part of the literature tends to raise attention towards the fact that energy communities could still be far from bringing more energy democracy and justice in this field. Another point to underline is that thanks to adopting a comparative perspective, this research also looks to identify how some traditional determinants of collective action initiatives as size but also socio-territorial context, can impact the capacity of these initiatives to bring more energy democracy and justice.
DEMOCRACY AND JUSTICE IN COLLECTIVE ACTION INITIATIVES IN THE ENERGY FIELD / A.j.s. Dudka ; tutor: L. Dorigatti ; coordinatore: G. Ballarino. Dipartimento di Scienze Sociali e Politiche, 2022 Dec 19. 34. ciclo, Anno Accademico 2021.
NOVEL APPROACHES IN GENETIC VARIANT DISCOVERY AND CLASSIFICATION TACKLING UNSOLVED PROBLEMS IN CANCER CLINICAL GENOMICS
Accurate interpretation of germline and somatic variants plays a pivotal role in clinical practice, providing the foundation for the correct diagnosis and targeted therapy in personalized medicine. However, variant classification is a complex process that requires the integration of multiple, often incomplete sources of information, prompting the development of bioinformatic tools that enable clinicians to prioritize mutations for validation and patient consulting. The primary objective of my PhD project is to identify and evaluate consistent methods for discovering mutations of scientific and clinical significance. To this aim, in 2020, we designed RENOVO, an algorithm harnessing machine learning. The algorithm was trained on the extensive ClinVar database and validated across multiple external databases. RENOVO exhibited high accuracy in classifying known pathogenic and benign variants. It provides a pathogenicity likelihood score, aiding in the interpretation of de novo or variants of unknown significance. Our comprehensive evaluation of RENOVO's predictions, spanning a four-year period, demonstrated its efficacy, as RENOVO correctly classified 82.6% of variants that underwent reclassification. This validation confirms RENOVO’s potential to predict future reclassification of variants currently deemed uncertain or conflicting in clinical significance. We applied variant interpretation tools, including RENOVO, to observational trials addressing clinical needs in clinical cancer genomics, on ovarian and breast cancers and myeloproliferative neoplasms (MPN) who did not previously show positivity for known MPN genetic drivers (“triple negative disease”).In the latter study, we found a high prevalence of mutations in a known cancer-associated gene, KMT2C. However, we noticed that KMT2C variants were highly dependent on the alignment algorithm used, due to high homology with a known pseudogene. Further investigation led to the discovery of a phenomenon that we named Genotype-Dependent Mismapping (GDM). Genomic regions subject to GDM are highly homologous regions that differ by one or few highly prevalent polymorphisms. As a consequence, the mappability of the region depends entirely on the polymorphic allele, which acts as a binary “switch”: one allele allows unambiguous mapping, whereas the other renders the region unmappable due to complete homology. We developed a tool to measure the probability of GDM for large genomic regions and we are currently applying our tool to existing studies to reassess the prevalence of known oncogenic variants. Overall, this project presents an effective strategy for addressing genomic challenges associated with variant detection, validation, and classification, emphasizing a patient-centric methodology.
NOVEL APPROACHES IN GENETIC VARIANT DISCOVERY AND CLASSIFICATION TACKLING UNSOLVED PROBLEMS IN CANCER CLINICAL GENOMICS / E. Bonetti ; tutor: L. Mazzarellaco-tutor: P. G. Peliccisupervisore: N. Soranzosupervisore: T. Pippuccicoordinatore: D. Pasini. Dipartimento di Oncologia ed Emato-Oncologia, 2024 Jan 21. 36. ciclo, Anno Accademico 2023/2024.
EFFICIENCY AND REALISM IN STOCHASTIC BANDITS
This manuscript is dedicated to the analysis of the application of stochastic bandits to the recommender systems domain. Here a learning agent sequentially recommends one item from a catalog of available alternatives. Consequently, the environment returns a reward that is a noisy observation of the rating associated to the suggested item. The peculiarity of the bandit setting is that no information is given about not recommended products, and the collected rewards are the only information available to the learning agent. By relying on them the learner adapts his strategy towards reaching its learning objective, that is, maximizing the cumulative reward collected over all the interactions.In this dissertation we cover the investigation of two main research directions: the development of efficient learning algorithms and the introduction of a more realistic learning setting. In addressing the former objective we propose two approaches to speedup the learning process. The first solution aims to reduce the computational costs associated to the learning procedure, while the second's goal is to boost the learning phase by relying on data corresponding to terminated recommendation sessions. Regarding the latter research line, we propose a novel setting representing use-cases that do not fit in the standard bandit model.
EFFICIENCY AND REALISM IN STOCHASTIC BANDITS / L. Cella ; tutor: N. Cesa-Bianchi ; coordinatore PhD program: P. Boldi. Dipartimento di Informatica Giovanni Degli Antoni, 2021 Jan 15. 33. ciclo, Anno Accademico 2020. [10.13130/cella-leonardo_phd2021-01-15].
INNOVATIVE SYSTEMS FOR THE MITIGATION OF RISKS: CATALYSTS AND SMART MATERIALS FOR DETECTION AND DECONTAMINATION OF HAZARDOUS CHEMICAL, BIOLOGICAL AND EXPLOSIVE SUBSTANCES
Incidents involving Chemical, Biological, Radiological, Nuclear and explosive agents (CBRNe), whether intentional or accidental, pose significant threats in the modern international scenario. The use of these substances, not only endangers human life, but also represents a threat to national defence, the economy and, above all, the environment due to their strong harmfulness, persistence and recalcitrance. Consequently, there is a critical need for the development of innovative systems to mitigate the risks associated with these hazardous agents. This research spans several areas related to the mitigation of chemical, and biological risks. The broad aim of this Ph.D. thesis was to overcome the problems and limitations currently present in some conventional decontamination and detection methods concerning these hazardous agents.Decontamination of Hazardous Biological Agents - In this field, it is a priority to focus on substances that are both environmentally and economically sustainable, as well as safe from a toxicity point of view, aiming to avoid the extensive use of chlorine-based solutions or to minimise the release of VOCs into the environment. In response to the COVID-19 pandemic, hydrogen peroxide has emerged as a sustainable alternative for inactivating SARS-CoV-2 in the liquid phase. A diluted 3% w/w H2O2 solution acidified to pH 2.5 by adding citric acid inactivated SARS-CoV-2 virus by more than 4 orders of magnitude in 5 min. Conversely, H2O2 solutions with no additives displayed a scarce virucidal activity, confirming that a pH-modifying ingredient is necessary to have a H2O2-based disinfectant active against this virus. Furthermore, recognizing that an acid and an oxidizing functionality are important for the inactivation of many pathogens, efforts were made to heterogenize and immobilize these functionalities within a system based on sulfonic acid cation exchange resins. After a simple contact treatment with an aqueous H2O2 solution, these resins demonstrated the ability to store oxidising species and release them over time. These systems exhibited potent biocidal efficacy against bacteria, resulting in a reduction of 9 and 5 orders of magnitude in Escherichia coli (Gram -) and Staphilococcus aureus (Gram +) viability, respectively, even after 1 min. Additionally, they showed effectiveness against viruses, with a complete absence of viral replication for SARS-CoV-2 (RNA virus), Herpes simplex (DNA virus), and Monkeypox (DNA virus) after 2 min. This drastic biocidal action is therefore attributable to the synergistic effect between the strong Brønsted acidity of the resins and the oxidising capacity due to the presence of H2O2 and in-situ formed peroxosulfonic species.Decontamination and abatement of hazardous chemicals - The transition from stoichiometric to catalytic methods is crucial for the effective and sustainable decontamination of hazardous chemicals. In this study, nanostructured porous materials, including commercial non-ordered aluminosilicates (SiO2-Al2O3), niobium oxide (Nb2O5), and modified synthetic saponites clays (NbSAP, Na-SAP, H-SAP), were selected and tested as heterogeneous catalysts for the liquid-phase degradation of two organophosphorus obsolete agrochemicals and CWAs simulants, namely paraoxon-ethyl and parathion-ethyl. Regarding synthetic saponite clays, they represent a class of hydrated smectite-type phyllosilicate materials, consisting of a 2:1 trioctahedral structure of alternating tetrahedral (T) and octahedral (O) sheets, typically composed of Si(IV), Al(III), Mg(II) and O2- sites, organised to form T-O-T layers alternating with an interlamellar space containing exchangeable cations and water molecules. Thanks to the ability to modulate the physico-chemical features of saponites at different levels: 1) by modifying the composition, structure, and morphology through varying the dilution of the synthesis gel in the hydrothermal
INNOVATIVE SYSTEMS FOR THE MITIGATION OF RISKS: CATALYSTS AND SMART MATERIALS FOR DETECTION AND DECONTAMINATION OF HAZARDOUS CHEMICAL, BIOLOGICAL AND EXPLOSIVE SUBSTANCES / S. Econdi ; tutor: A. Caselli ; co-tutor: M. Guidotti ; coordinatore: D. Roberto. Dipartimento di Chimica, 2024 Feb 27. 36. ciclo, Anno Accademico 2023.
A COMPREHENSIVE OVERVIEW OF PSYCHOLOGICAL AND ETHICAL IMPLICATIONS ON THE ROLE OF CAREGIVER IN THE CLINICAL PATHWAY OF PATIENTS NEWLY DIAGNOSED WITH BREAST AND PROSTATE CANCER
In recent years, research on the caregivers’ involvement in the Shared Decision-Making and cancer care has burgeoned in the psychological field; however, knowledge gaps still exist because most studies have failed to acknowledge the multidimensional nature of agreement/disagreement among dyads in the Shared Decision-Making and cancer trajectory, especially during the early phase of the disease (diagnosis and starting of treatments). My Ph.D. thesis focused on the agreement among patient-caregiver dyads along the cancer care pathway, from the diagnosis onward, and their mutual influence on the psychological adjustment to cancer in breast and prostate cancer patients and their caregivers. Along the cancer trajectory, dyads may experience episodes of conflicts, so it has been evaluated the concept of conflict from an ethical standpoint. The current projects highlighted the importance and need for a multidisciplinary approach to dyads, involving physicians, psycho-oncologists, and clinical ethicists. A holistic approach may help patients and caregivers coping with cancer, being supportive to each other and expressing their preferences, fears, and needs. Specifically, four studies addressed the dyadic agreement in the Decision-Making and the interdependence among dyads from a psychological standpoint through quantitative and qualitative methods. In particular, the first study explored the preferences of breast and prostate cancer patients and their caregivers in the Shared Decision-Making. Given the relevance of caregivers’ involvement in the Decision-Making, two studies demonstrated the importance of a renovated family-centered model of cancer care and the need of structuring psychological interventions tailored on the dyads or on the single member. Specifically, although patients often have at least one caregiver by their side, who provide practical and emotional support, it is unclear whether dyads establish their roles and the level of involvement in the Decision-Making process. Indeed, two studies showed that most dyads disagree on the preferences and actual involvement in the Decision-Making. Moreover, caregivers are not just a supporter, as she experiences the disease almost as much as the patients. Dyadic members experience high distress and influence each other’s adjustment to cancer. Findings from the qualitative assessment support the idea of cancer as a family disease. Dyads described the cancer diagnosis as an expected and shocking event, that needs to be addressed together as soon as possible. However, patients and caregivers experience different concerns: patients’ worries refer to the cancer recurrence and the body image, while caregivers’ worries about their ability to support patients. Then, in order to evaluate the caregivers’ involvement at a multilevel, two studies assess the concept of conflict from an ethical standpoint. The first study proposed a taxonomy of the moral conflict that may emerge along the cancer trajectory. Finally, a case report provides the basis for future investigations about the presence of moral conflicts even in the early stage of cancer (diagnosis). In conclusion, findings of this PhD projects suggest the importance of caregivers’ involvement along the cancer care pathway and the need to promote psychological interventions both for patients, caregivers, and the dyads. Finally, the present thesis gives future directions to increase breast and prostate cancer dyad well-being, reducing the Fear of Cancer Recurrence.
A COMPREHENSIVE OVERVIEW OF PSYCHOLOGICAL AND ETHICAL IMPLICATIONS ON THE ROLE OF CAREGIVER IN THE CLINICAL PATHWAY OF PATIENTS NEWLY DIAGNOSED WITH BREAST AND PROSTATE CANCER / C. Cincidda ; added supervisor: S. Oliveri ; internal advisor: V. Sanchini ; tutor: G. Pravettoni ; phd coordinator: S. Minucci. Dipartimento di Oncologia ed Emato-Oncologia, 2023 Dec 12. 35. ciclo, Anno Accademico 2023.
NF-YA ISOFORMS ROLES IN CLAUDIN LOW CANCERS
NF-Y is a pioneer transcription factor that regulates the expression of genes involved in cell cycle progression and metabolism. It is composed of: NF-YA, NF-YB and NF- YC subunits. NF-YA recognizes and binds to the CCAAT box, a DNA element found enriched in the promoters of cancer-specific genes in several tumor gene expression profiles analyzed, including those of breast, colon, thyroid, prostate, and leukemia cancers. NF-YA is present in two major isoforms called NF-YAl (long), and NF-YAs (short), which alternative splice exon 3. NF-YAl/NF-YAs ratio changes in different tissues, cell lines and differentiational stages. A robust increase in mRNA levels of NF- YA has been observed in different tumors of epithelial origin, among which BRCA and STAD. In human breast and gastric cancers and cell lines, NF-YAs isoform is highly expressed in all subtypes except in Claudinlow in which NF-YAl isoform is predominant. Recently it has been identified a 158 genes signature defining Claudinlow tumor subclass common to BRCA and STAD, correlating with high NF-YAl/NF-YAs ratio.For the first part of my PhD project, I worked with two different BRCA Claudinlow cell lines that mainly express NF-YAl. Through CRISPR/Cas9n genome-editing I obtained homozygous exon 3 deleted clones (NF-YAl-KO). NF-YAl-KO clones had an impaired ability to form compact aggregates, their migration, extravasation, invasion, and metastasis formation abilities were significantly impaired both in vitro and in vivo compared to negative controls. RNA-seq data underlined pathways downregulated in all NF-YAl-KO clones, among which EMT and angiogenesis, corroborating in vitro and in vivo data.In parallel, I have investigated the role of different RNA-binding proteins in driving NF-YA alternative splicing, among which RBFOX2, QKI and CELF2 both in BRCA and in STAD cell lines. Through transient overexpression, I validated RBFOX2 as splicing factor promoting the maturation of NF-YAl.Finally, through a frameshift mutation in NF-YA gene, I have investigated the effect of NF-YA alternative translation initiation sites (TISs) usage in HeLa cells and its impact at cellular and molecular levels.
NF-YA ISOFORMS ROLES IN CLAUDIN LOW CANCERS / M. Londero ; tutor: R. Mantovani. Dipartimento di Bioscienze, 2024 Mar 08. 36. ciclo, Anno Accademico 2023.
DIFFERENT TECHNIQUES FOR DENTAL IMPLANTS SITE PREPARATION: A CONTROLLED CLINICAL TRIAL
Dental implants should be considered a highly predictable long-term and evidence-based treatment option for single, partial, and complete edentulism of the jaws, as indicated by numerous scientific articles. The primary aim of the present study was to determine whether the application of one implant site preparation protocol (“Step drilling sequence”) could lead to a significantly lower marginal bone resorption as compared to another protocol (“Straight drilling sequence”) at 1-year follow-up. The secondary aims were to evaluate if the tested protocol (Step drilling sequence) could modify the clinical and/or patient-centered variables We included and treated 60 subjects, for a total of 86 implants. We found no evidence of a difference in bone resorption, which increased significantly over time, between the two groups. Regarding the secondary outcomes, no statistically significant difference was observed between the test group and the control group, neither for peri-implant probing depth (PD), for bleeding index (BI), plaque index (PI) nor for the quality of life. In conclusion, from a purely biological perspective, there is currently no evidence in favor of using a step drill over a traditional straight drill. However, from a clinical standpoint, the use of step drills is advantageous for operator convenience compared to traditional drills.
DIFFERENT TECHNIQUES FOR DENTAL IMPLANTS SITE PREPARATION: A CONTROLLED CLINICAL TRIAL / B. Morandi ; tutor: S. Taschieri ; coordinatore: M. Del Fabbro. - Milano. Dipartimento di Scienze Biomediche, Chirurgiche ed Odontoiatriche, 2024. 36. ciclo, Anno Accademico 2023.
MASS SPECTROMETRIC STRATEGIES TO IDENTIFY BIOACTIVE COMPONENTS FROM NATURAL EXTRACTS WITH ANTI-INFLAMMATORY ACTIVITIES
Oxidative stress and chronic inflammation are two conditions that can worsen various diseases, spanning metabolic, neurological disorders to tumour formation. In this context, a wide source of bioactive molecules exhibiting anti-inflammatory effects, combined with antioxidant potential, can be found in fruit and vegetables extracts. In particular, it has been reported that certain phytoconstituents are able to target two closely related systems within the cell, namely the NRF2 and NF-κB pathways. While the NRF2 pathway is responsible for regulating an extensive panel of antioxidant enzymes, NF-κB is the main effector pathway involved in inflammation. Phytochemicals in general mediate their effects by modulating the DNA-binding capacity of both these transcriptional factors and, in turn, correcting an unbalanced cellular state related to an uncontrolled inflammatory condition. Whereas previously, as a source of these phytochemicals, the focus was on the 'nobler' parts of the plant (i.e. the fruits, leaves and roots), today much more emphasis is placed on the analysis of industrial by-products. Specifically, this strategy aimed at the reuse of industrial by-products could reduce the impact of cultivation on waste production, minimizing disposal problems while obtaining a potential source of bioactive compounds. Even though there is general knowledge on the possible targets of these compounds, newer techniques combining different -omics approaches could better illustrate the biological impact of these extracts while identifying the phytochemical compounds responsible for it. Combining these different approaches to a concept of circular economy to valorise waste products could be a key to obtain useful health care products to prevent or even treat oxidative and inflammatory conditions.In particular, in this thesis, I will explore the antioxidant and anti-inflammatory activities of different extracts obtained from by-products of grape and apple cultivation. This investigation will employ a multifaceted approach, integrating engineered cell cultures for phenotypic screening of bioactive extracts, Mass Spectrometric-based metabolomics to unravel the primary components of these extracts, and Mass Spectrometric-based proteomic studies to comprehensively analyse their impact on the cell proteome. The exploration of potential mechanisms of action will be conducted through both in-vitro and ex-vivo quantitative proteomic studies. Conclusively, the thesis will showcase the application of a MS-CETSA (Mass Spectrometry – Cellular Thermal Shift Assay) methodology to probe the direct target of a bergamot leaves polyphenols extract.
MASS SPECTROMETRIC STRATEGIES TO IDENTIFY BIOACTIVE COMPONENTS FROM NATURAL EXTRACTS WITH ANTI-INFLAMMATORY ACTIVITIES / G. Ferrario ; tutor: G. Aldini ; coordinatore: G. Vistoli. Dipartimento di Scienze Farmaceutiche, 2023 Feb 13. 36. ciclo
EFFECTIVENESS OF SUGAR-FREE CHEWING GUM CONTAINING PROBIOTICS ON ORAL HEALTH
Background: Heyndrickxia coagulans is a spore-forming probiotic increasingly investigated for oral health applications due to its stability, antimicrobial activity, and ability to modulate the oral microbiota. Chewing gum has recently emerged as an innovative delivery system that may enhance probiotic dispersion and contact with oral surfaces. To comprehensively evaluate the oral health potential of H. coagulans, a four-phase research program was conducted encompassing: (1) a systematic review and meta-analysis of its clinical evidence; (2) a kinetic study assessing salivary persistence following gum administration; (3) a randomized clinical trial evaluating plaque colonization and microbiome modulation; and (4) a randomized controlled trial testing its efficacy against halitosis.Methods: Phase 1 systematically reviewed randomized and non-randomized clinical studies up to September 2025, extracting data according to PRISMA guidelines and assessing risk of bias with RoB 2.0 and ROBINS-I. Meta-analysis focused on salivary Streptococcus mutans. Phase 2 was a randomized crossover study in 10 adults who chewed probiotic gums containing H. coagulans SNZ1969® or Lacticaseibacillus rhamnosus GG (microencapsulated or not). Saliva was sampled over 2 hours to quantify CFUs and determine kinetic profiles. Phase 3 was a double-blind, placebo-controlled trial in 52 adults consuming H. coagulans gum (5×10⁸ CFU/pellet) or placebo for four weeks. Dental plaque collected at T₀–T₃ underwent qPCR for H. coagulans and 16S rRNA sequencing for microbiome analysis. Phase 4 was a parallel double-blind RCT in the same cohort assessing halitosis outcomes. Volatile sulphur compounds (VSCs)—hydrogen sulphide (H₂S) and methyl mercaptan (CH₃SH)—were measured using OralChroma™ at four time points.Results:Phase 1 included eight studies (n=30–183 participants). H. coagulans significantly reduced salivary S. mutans (pooled effect size −0.99; p<0.01) and improved gingival and periodontal parameters, although heterogeneity was high. Phase 2 showed that all probiotics peaked in saliva at T₁, with H. coagulans achieving the highest early concentrations (T₁–T₃; mean 6.1–5.6 log₁₀ CFU/mL). Probiotics remained detectable up to two hours post-chewing. Phase 3 demonstrated that H. coagulans was undetectable at baseline and in all placebo samples, but appeared in 71.4% of subjects at T₁ and 61.9% at T₂, with persistence in 9.5% at T₃. Microbiome analysis revealed transient depletion of dysbiosis-associated genera (Prevotella, Tannerella, Treponema, Neisseria, Leptotrichia) and selective enrichment of commensal taxa. Phase 4 found that both groups showed progressive VSC reductions, but declines were more pronounced with the probiotic. H₂S decreased from baseline at T₂ (p=0.008) and T₃ (p=0.031), with median levels approaching zero, although between-group differences were not significant. CH₃SH showed no significant changes.Conclusions: Across all four phases, H. coagulans demonstrated promising oral health potential. It reduced cariogenic bacteria, persisted transiently in saliva, reached and modulated the dental biofilm, and significantly lowered hydrogen sulphide levels in vivo. Effects were modest, strain- and host-dependent, and more consistent for H₂S than for CH₃SH. Chewing gum proved to be an effective delivery vehicle. Larger, longer, and mechanistically oriented trials are warranted to better define the therapeutic role of H. coagulans in preventive and adjunctive oral care.
EFFECTIVENESS OF SUGAR-FREE CHEWING GUM CONTAINING PROBIOTICS ON ORAL HEALTH / S. Cirio ; tutor: M.G. Cagetti ; coordinatore: M. Del Fabbro. - Aula Devoto. Dipartimento di Scienze Biomediche, Chirurgiche ed Odontoiatriche, 2025 Dec 19. 38. ciclo, Anno Accademico 2024/2025.
L¿ANNESSIONE DELLA CRIMEA ALLA FEDERAZIONE RUSSA NELLE PAROLE DELLA STAMPA. PRATICHE DISCORSIVE E COSTRUZIONE DELL¿IDENTITÀ NAZIONALE
The mass media play a central role in contemporary societies, being the news a fundamental actor in shaping our identities as citizens and our worldviews. Through the stories and representations built by journalists, public opinion produces and reproduces a shared cognition of reality.The main argument of this doctoral thesis is Crimea’s annexation to the Russian Federation as it was represented by some of the prominent Russian national newspapers. Crimea became a Russian republic in March 2014 following a bitterly contested by the international community referendum, which generated a crisis in relations between Russia and the West still far away from being resolved.The aim of the thesis is two-fold. The first one is to analyze the narrative and linguistic strategies through which discourses of the annexation, along with their cultural and ideological implicatures, have circulated through the print media. The second goal is to verify whether and to what extent it is possible to identify alternative and competing narratives in the Russian press. To the purpose, a corpus of texts has specifically been prepared. The corpus consists of one hundred and eighty news and commentary articles retrieved from six national newspapers. The newspapers selection was based on popularity and circulation criteria. The analysis involves three dimensions: the representation of national identity, the representation of social actors and the representation of reported speech. The methodology adopted draws from Critical Discourse Analysis, Discourse Historical Approach and the model for the representation of social actors developed by Theo van Leeuwen (1996).Overall, a considerable degree of uniformity characterizes the newspapers analyzed. Due to their loyalty to the Kremlin, Izvestija, Rossijskaja Gazeta, Komsomol’skaja Pravda, and – to some extent – Kommersant ‘’ reproduced or even anticipated, the discourses circulated by the political elite. Novaya Gazeta and Vedomosti take a different line, albeit with distinct nuances, consistent with their ideological position as part of the so-called systemic opposition. Regarding the representation of national identity, the whole discourse was built on the polarization between the Russian nation and an external enemy, namely all those who do not recognize Russia the status of a ‘Great Nation’. In this ‘Great Nation’ conceptualization, a foreground position is given to ethnic belonging, which is made to coincide with linguistic identity. The discursive strategies employed by journalists can be summarized as follows: emphasis on the difference between ‘us’ and ‘them’, emphasis on the alleged historical similarity between the present time and the Second World War, black and white description of the enemy, delegitimization of the Ukrainian state, shift of blame and responsibility by reversing the victim/culprit scheme.Concerning the second dimension – the representation of social actors – the strategies and narrative schemes adopted in no small extent duplicate the representation of the national identity. The discourse is articulated around an intense polarization between an in-group, namely the ethnic Russians, and an out-group, namely all the rest. An example of such polarization is the use of epithets drawing from the memory of the WWII – in particular banderovcy, and opolcency -, to leverage the emotional sphere of public opinion and to exclude from the discourse any possible articulated positions, distinctions, or identities that eschew binary representation.The third analytical dimension has highlighted to what extent reported speech constitutes a significant issue in many texts. The preferred tendency is that of direct quotation, isolated from the text by graphic signs such as quotation marks or hyphens. A second element is the tendency to favour neutral verba dicendi to introduce speakers’ utterances: zajavit ‘, skazat’, ob’jasnit’ are those
L¿ANNESSIONE DELLA CRIMEA ALLA FEDERAZIONE RUSSA NELLE PAROLE DELLA STAMPA. PRATICHE DISCORSIVE E COSTRUZIONE DELL¿IDENTITÀ NAZIONALE / F. Volpi ; tutor: R. Vassena; coordinatore: M. V. Calvi. Dipartimento di Lingue e Letterature Straniere, 2021 Mar 29. 33. ciclo, Anno Accademico 2020. [10.13130/volpi-francesca_phd2021-03-29].
NUTRITION AND IMMUNITY: MOLECULAR APPROACHES
Nutrition plays an important role in modulating livestock species immunity. Therefore, this thesis aimed at evaluating the effects of different dietary molecules used in animal nutrition on mammalian and avian immunity. Both, in vitro functional analyses and OMIC technologies (proteomics and miRNAomics) were implemented herein for an integral characterization of the molecules’ impact on the animals’ immune response. Specifically, in this thesis the in vitro impact of the n-6 conjugated linoleic acid (CLA), citrus pectin (CP), and porcine milk exosomes and n-3 polyunsaturated fatty acids (PUFA) on bovine, chicken and porcine mononuclear cells immune response was evaluated, respectively.In the first study, the in vitro activity of CLA on bovine monocytes apoptosis and immune activities, including chemotaxis, phagocytosis, killing capability, and extracellular reactive oxygen species (ROS) production was assessed. Anti-apoptotic effects and an increase in extracellular ROS production during experimental pro-inflammatory conditions were observed, only when using the mixture of the two main isomers of CLA in equal proportions (50:50). The present results demonstrated for the first time that CLA does have immunomodulatory effects on some functions of bovine monocytes in vitro and that the CLA (50:50) mixture is more effective than the CLA isomers individually. The proteomics analysis performed on bovine peripheral blood mononuclear cells (PBMC) revealed that CLA (50:50) mixture does modulate bovine PBMC proteome, supporting the antiapoptotic and immunomodulatory effects observed in the previous in vitro study on bovine monocytes, and propose a potential cytoprotective role of CLA (50:50) mixture against oxidative stress.In the second study, the in vitro activity of CP on chicken monocytes viability, apoptosis, chemotaxis and phagocytosis was assessed. The study demonstrated for the first time that CP inhibits monocytes’ chemotaxis and phagocytosis in vitro, suggesting a potential anti-inflammatory activity. The proteomics analysis carried out on chicken PBMC provided a proteomics background to the anti-inflammatory activity of CP, demonstrating that the in vitro reduction of phagocytosis and chemotaxis is associated with changes in proteins related to the actin cytoskeleton.In the third study, the in vitro activity of porcine milk exosomes on porcine monocytes viability, apoptosis, chemotaxis, phagocytosis, killing capability and extracellular ROS production was assessed. Milk exosomes were successfully purified from sows’ milk and characterized using their size, concentration, morphology, and exosome protein markers. This study reported for the first time that porcine milk exosomes can be internalized by porcine monocytes in vitro and that they can modulate the cell's immune response, by decreasing their chemotaxis and phagocytosis; and increasing their ROS production under resting and pro-inflammatory conditions. The proteomics analysis performed on porcine PBMC demonstrated for the first time that porcine milk exosomes can modulate porcine PBMC proteome in vitro. Moreover, the gene ontology (GO) functional analyses revealed that porcine milk exosomes enrich biological processes related to innate immune-related processes and exosome uptake processes, supporting the immunomodulatory effects and the exosome internalization observed in the previous in vitro study.In the last study, the in vitro activity of the n-3 PUFA, docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), on porcine monocytes viability, apoptosis, chemotaxis, phagocytosis, and intracellular, extracellular and total ROS production was assessed. The results of the study showed that DHA and EPA at the highest concentration (200 µM) decreased porcine monocytes' viability. In addition, it was reported for the first time that DHA and EPA can exert differential in vitro immunomodulatory effects in pigs, by dam
NUTRITION AND IMMUNITY: MOLECULAR APPROACHES / G.d.l.a. Avila Morales ; tutors: F. Ceciliani, A. Sánchez Bonastre ; supervisor: M. Bonnet, F. Ceciliani. Università degli Studi di Milano, 2022 Sep 28. 34. ciclo, Anno Accademico 2021.
NEW ORGANIC AND HYBRID INORGANIC/ORGANIC LUMINESCENT MATERIALS
Triimidazo[1,2-a:1’,2’-c:1”,2”-e][1,3,5]triazine, is a molecule which has been studied by Cariati et. al. since 2017. This system possesses both Aggregation Induced Emission (AIE) and Room Temperature Ultralong Phosphorescence (RTUP) properties, whose discovery fueled further derivatization of the core: in fact, bio-active products, metal complexes and coordination polymers (CP), and phosphorescent organic emitters were synthesized and characterized. My work focused on a further derivatization of the TT scaffold with selected pendants, aiming at obtaining RTP materials and suitable ligands to be employed in luminescent metal complexes and coordination polymers.
NEW ORGANIC AND HYBRID INORGANIC/ORGANIC LUMINESCENT MATERIALS / D. Maver ; tutor: E. Cariati ; co-tutor: A. Colombo ; coordinatore: D. Roberto. - Via Golgi 19, Milano. Università degli Studi di Milano, 2025 Feb 04. 37. ciclo, Anno Accademico 2024.
A PHASE III PROSPECTIVE RANDOMIZED TRIAL TO EVALUATE THE IMPACT OF AUGMENTED REALITY DURING ROBOT-ASSISTED RADICAL PROSTATECTOMY ON THE RATES OF POSTOPERATIVE SURGICAL MARGINS
Background In contemporary series, the rate of positive surgical margins (PSMs) at final pathology after robot-assisted radical prostatectomy (RARP) spans from 6.5 to 32%. Novel technologies are urgently required for implementing preservation of neuro-vascular bundles, while simultaneously decrease the rates of PSMs. 3D virtual reconstructions of 2D cross-sectional imaging have been increasingly adopted to facilitate the surgeon in better understanding the surgical anatomy. Despite initial promising results, as of today, no prospective studies were conducted to validate these technologies in daily practice. Moreover, additional comprehensive data are required to test the use of ex-vivo fluorescence confocal microscopy (FCM) as a valid alternative to intraoperative frozen section analysis (IFS) for real-time margin evaluation.Methods This is a Phase III, monocentric, prospective and randomized trial that compares augmented reality (AR) RARP with standard approach (standard RARP) for reducing the rates of PSMs. A total of 318 patients will be randomized in a 1:1 ratio to: -AR RARP: prostate 3D reconstruction according to preoperative multiparametric magnetic resonance imaging (mpMRI). The model will be projected in the surgical field using the TileProTM technology. After prostate extraction, the virtual 3D model of the prostate will be overlapped to the gland with the used of mixed reality (Hololens glasses, Microsoft, USA) and margins will be inked according to the 3D model. Intraoperative margins will be analyzed by both IFS and FCM (Vivascope 2500M-G4);-standard RARP: nerve-sparing approach according to mpMRI and mpMRI-guided standard IFS.Primary outcome is the percentage of PSMs between the two groups, with several subgroup analyses performed according to tumor and mpMRI characteristics. Secondary outcomes are: 1) the rate of nerve-sparing approaches within the two groups; 2) 3-, 6- and 12-months recovery of erectile function (IIEF-5 score) after surgery; 3) accuracy of the Vivascope 2500M-G4 vs. IFS in detecting PSMs.We reported results of the ad-interim analysis, that was planned after recruiting 159 patients.Results Overall, 80 (51%) vs. 79 (49%) patients were treated with, respectively, standard RARP vs. AR RARP. Twenty seven (33.8%) vs. 15 (19%) patients in standard RARP vs. AR RARP groups had PSMs at final pathology (p=0.01). Subgroup analyses showed a trend for lower PSMs in the AR RARP group across all tumor and mpMRI variables, with the exception of those lesions located in the apex of the prostate. This advantage was related to higher rates of margins positivity at IFS analysis in AR RARP vs standard RARP (38 vs. 20%; p=0.01). Median (IQR) surgical operative time was 202 (181-223) vs. 209 (190-239) mins for, respectively, standard RARP vs. AR RARP (p=0.17). No differences in final right (p=0.9) or left (p=0.3) nerve sparing approaches were observed between the two groups. No differences in biochemical recurrence (BCR) or erectile function recovery over time were recorded. At ex-vivo FCM, the inter-observer agreement between pathologists ranged from moderate (k=0.74) to almost perfect (k=0.90). For a highly experienced pathologist, a good balance among sensitivity (70.5%) specificity (91.8%), PPV (80.0%) and NPV (87.1%) was reached when analysis ex-vivo FCM images. Last, the agreement between ex-vivo FCM and IFS analysis ranged from moderate (k=0.62) to strong (k=0.86). Conclusions At the time of the ad interim analysis of this Phase III prospective randomized trial, we already observed lower rates of PSMs in patients treated with AR RARP vs. standard RARP. This observed advantage applied to all patient subgroups and did not compromise oncological control of the disease nor erectile function recovery. Moreover, prostate margins evaluation at ex-vivo FCM appears to be feasible and reliable, with a good agreement between readers. Nevertheless, as of today, the performance of FCM see
A PHASE III PROSPECTIVE RANDOMIZED TRIAL TO EVALUATE THE IMPACT OF AUGMENTED REALITY DURING ROBOT-ASSISTED RADICAL PROSTATECTOMY ON THE RATES OF POSTOPERATIVE SURGICAL MARGINS / S. Luzzago ; tutor: N. Fusco ; Phd Coordinator: C. Sforza, M. Clerici ; supervisor: G. Musi. Dipartimento di Oncologia ed Emato-Oncologia, 2025 Jan 23. 37. ciclo
DEVELOPMENT OF NEW PEPTIDE-BASED NANOMATERIALS AS POTENTIAL TOOLS FOR CRYOPRESERVATION OF TISSUES AND ORGANS
Transplant practise has improved life duration and quality of many patients. However, the worldwide need is not met yet due to the limited number of donor organs and the inefficient preservation methods, that hinder their long-term preservation. The main obstacle to achieve a successful cryopreservation is represented by the freezing and thawing processes needed to achieve cryogenic temperatures that can cause permanent and irreversible cellular damages. The solution to this issue may be found in nature, as several organisms living in extreme conditions produce specific proteins, termed antifreeze (glyco)proteins (AF(G)Ps), that are able to depress the freezing point of body fluids, control the ice crystal nucleation and inhibit their growth. In particular, the latter AFP property, the ice recrystallization inhibition (IRI), is highly desirable for new synthetic antifreeze materials as ice recrystallization during thawing is associated to cellular damage during cryopreservation. Therefore, one of the main challenges in biomedical research for organ transplantation is finding non-toxic and biocompatible antifreeze compounds that enable subzero storage and cryopreservation without causing tissue damage. As the synthesis of molecules to be used for cryopreservation represents an urgent need, in the last decades researchers have been extensively focused on it. Since the production of AF(G)Ps is expensive and presents several difficulties in its scale-up, and considering their potential cytotoxicity/immunogenicity and their tendency to denature, efforts have been focused on synthesising macromolecular cryoprotectant-polymers or minimising the AFP primary sequence through the synthesis of peptides or lastly, increasing the antifreeze activity through the exploitation of a multimeric approach, taking advantage from self-assembly processes or the simultaneous presentation of several ice binding motifs, that allows to reduce the concentration required for the effect. Although extensive efforts have been devoted to the synthesis of novel peptide or peptide-based cryoprotectants, limited results were achieved probably due to the poor understanding of the AF(G)Ps mechanism of action at molecular level and the lack of comprehensive structure-activity relationships. Considering the relevance and the global impact of that research field, finding new effective antifreeze compounds that can be translated into clinic represents an urgent need.Being inspired by the multimeric approach, the aim of this PhD project was to develop new peptide-capped materials possessing an antifreeze activity by conjugating the N-terminus of peptides derived from type I AFP HPLC6 to different moieties that are known for their self-assembling ability. The goal of decorating nanoparticles (NPs) with peptides relies on combining the ice binding capability of peptides with the size of a NP, mimicking the globular protein bulkiness. In order to reach the goal, different approaches were exploited, as i) the on-resin peptide functionalization with lauric acid, ii) the in-solution peptide functionalization with the FDA approved polymer poly(lactide-co-glycolide) (PLGA) or iii) to gold nanoparticles (AuNPs), by inserting in peptide sequence a thiol-bearing binding functionality, and lastly, iv) the on-resin peptide functionalization with Aggregation-Induced Emission (AIE) luminophores, organic molecules characterized by uniquely fluorescent properties in the aggregate or solid state. First of all, the synthesis of the precursor peptides has been optimized and their secondary structure has been characterized by circular dichroism (CD) and Attenuated Total Reflection-Fourier Transform Infrared (ATR-FTIR) spectroscopy. Considering the presence of different conformations and the lack of a preferred helical one, the secondary structure of the minimised sequence of type I AFP HPLC6 was stabilised using a click chemistry reaction, the copper(I)-catalysed azide-alkyne cycload
DEVELOPMENT OF NEW PEPTIDE-BASED NANOMATERIALS AS POTENTIAL TOOLS FOR CRYOPRESERVATION OF TISSUES AND ORGANS / E. Impresari ; tutor: S. Pellegrino ; coordinatore: G. Vistoli. Dipartimento di Scienze Farmaceutiche, 2023 Mar 27. 35. ciclo, Anno Accademico 2022.
A MULTILEVEL APPROACH TO DEFINE THE ROLE OF IMMUNE AND METABOLIC CUES ON THE MACROPHAGE/FIBROBLAST INTERPLAY DURING FIBROSIS DEVELOPMENT
Alterations in tissue homeostasis occur during inflammation, oxygen deprivation, and remodeling, but they are generally reversible processes. When the cause of injury is severe, persistent or repetitive, acute inflammation could become chronic and dysregulated wound-healing could lead to fibrosis. Dynamic interplay through immune and non-immune cells, such as macrophages and tissue fibroblasts, is crucial to determinate pathological outcome. Macrophages (Mφ) are highly plastic cells, able to assume different functional phenotypes depending on the microenvironment. Fibroblasts (Fb) are heterogeneous cells that can be activated into myofibroblasts, which are the main sources of extracellular matrix (ECM) components. In inflamed tissues and fibrotic scars macrophages accumulate in areas of hypoxia, which is known to impact on macrophages activation. In these contexts the interplay of macrophages with fibroblasts likely affects their biology. In this study, we establish an in vitro direct contact co-culture model with macrophages and fibroblasts in order to elucidate cell-to-cell interactions. The system set-up take into account different microenvironment alterations, including T-cells cytokine secretion and oxygen deprivation, in order to mimic adaptive immune contribution and metabolic switch during inflammation and fibrosis. To investigate the relevance of different microenvironmental cues on Mφ and Fb activation, we stimulated the co-cultures with pro-inflammatory stimuli (LPS+IFNor with pro-fibrotic cytokine (IL-4) or with 1% of oxygen tension (to mimic severe hypoxia). Model parameters will be extracted from molecular profiling approaches investigating 44 different combinations of Mφand Fb polarized into inflammatory (MI and FbI) and fibrotic (MF and FbF) settings under normoxic and hypoxic conditions, in single cell cultures and direct-contact co-cultures. A multi-level strategy let us to compare different samples, to discriminate the effect of single variable on the system and to combine up to four variables together. We found that, when cells are in direct-contact in hypoxic environment, resting and pro-inflammatory Mφ and Fb assume or maintain an enriched inflammatory signature whereas pro-fibrotic macrophages inhibit the acquisition of a pro-inflammatory phenotype of fibroblasts. Indeed, single influencing factor alone is not able to induce differences in resulting phenotype but when immune stimuli, hypoxia and co-cultivation are combined together, for a long period of time, they change Mφ and Fb transcriptional landscape. Implementation of these findings with functional assays is essential to deeper investigate this crosstalk in chronic inflammation and fibrosis to translate candidate genes into predictive biomarkers.
A MULTILEVEL APPROACH TO DEFINE THE ROLE OF IMMUNE AND METABOLIC CUES ON THE MACROPHAGE/FIBROBLAST INTERPLAY DURING FIBROSIS DEVELOPMENT / E. Setten ; supervisor: M. Locati. DIPARTIMENTO DI BIOTECNOLOGIE MEDICHE E MEDICINA TRASLAZIONALE, 2020 Jan 31. 32. ciclo, Anno Accademico 2019. [10.13130/setten-elisa_phd2020-01-31].
ADVANCES IN STOCHASTIC ANALYSIS ON SPACES OF MEASURES: KOLMOGOROV EQUATIONS RELATED TO STOCHASTIC FILTERING AND MEAN FIELD OPTIMAL STOPPING
The purpose of this Thesis is to study some problems arising in the context of stochastic analysis and stochastic optimal control, where some relevant variables take values in the space of positive or probability measures.Most of the work is dedicated to the introduction and study of some backward Kolmogorov equations on spaces of measures associated to filtering problems. Measure-valued processes arise naturally in the context of stochastic filtering and one can formulate two stochastic differential equations, called Zakai (Z) and Kushner-Stratonovich (KS) equations, that are solved by a positive and a probability measure-valued process respectively. A classical way to study these problems is to assume that the measure-valued processes admit a density and then exploit stochastic calculus in Hilbert spaces. The approach used in this Thesis differs from this since we do not assume the existence of a density and we work directly in the context of measures. We formulate two backward Kolmogorov equations, which are parabolic partial differential equations with a given final condition, one over a space of positive (in the Z case) and one over a space of probability (in the KS case) measures. In the recent literature, PDEs on spaces of probability measures have been a topic of great interest, thanks to the connection with mean field games and McKean-Vlasov equations. Here we state a new equation on a space of probability measures, obtained in a completely different framework, and we introduce a PDE on a space of positive measures, which is one of the first problems of this type. We study the existence and uniqueness of a solution from two different points of view.First, we focus on classical solutions. We prove that both the equations introduced admit a unique classical solution, as long as the final condition is chosen regular enough. To achieve this, we need some intermediate results of independent interest. In particular, we prove Itô formulas for the composition of filtering processes and real-valued functions defined on the space of measures. Moreover, we study the regularity of the solution to the filtering equations with respect to the initial datum. Since we are dealing with functions over spaces of measures, a key point is to discuss proper notions of derivatives, especially in the case of positive measures. After this, we investigate the case when the final condition is less regular. A first remark is that the approach we used in the classical case strongly depends on the regularity of the final condition. Thus, we need to change the notion of solution and look for viscosity solutions. To this aim, we focus on the PDE associated to the KS equation. In the literature, only a few results are available on viscosity solutions for this kind of problem, and in particular, uniqueness is a very challenging issue. In this case, we prove the existence and uniqueness of a viscosity solution and, in particular, we provide a comparison theorem. In the final part of this Thesis, we deal with a different problem, which arises in the context of optimal stopping theory. We study a class of finite horizon time-inconsistent optimal stopping problems (OSPs) of mean field type, which includes those related to mean field diffusion processes and recursive utility functions. The mean field interaction is due to the fact that we consider a terminal cost depending not only on the stopped process but also on its law (which is different from the law of the process evaluated at the stopping time).Despite the time-inconsistency of the OSP, we show that it is optimal to stop when the value-process hits the reward process for the first time, as is the case for the standard time-consistent OSP. We solve the problem by approximating the corresponding value-process with a sequence of Snell envelopes of processes, for which a sequence of optimal stopping times is constituted of hittin
ADVANCES IN STOCHASTIC ANALYSIS ON SPACES OF MEASURES: KOLMOGOROV EQUATIONS RELATED TO STOCHASTIC FILTERING AND MEAN FIELD OPTIMAL STOPPING / M. Martini ; tutor: M. A. Fuhrman ; coordinatore dottorato: D. P. Bambusi. Dipartimento di Matematica Federigo Enriques, 2023 Mar 02. 35. ciclo, Anno Accademico 2022.
LIPIDOMICS UNVEILS THE ROLE OF BIOACTIVE LIPIDS IN CELLULAR MODELS OF CYSTIC FIBROSIS
Cystic fibrosis is a genetic disease affecting mainly the lungs, causing chronic inflammation and recurrent infections. The lipid accrual directly contributes to the persistent damage in the cystic fibrosis airways; thus, the deregulated lipid metabolism can be targeted with pharmacological treatment. Here, we aim at demonstrating, in an in vitro model of cystic fibrosis, the role of inflammatory and bioactive lipids in the disease pathophysiology and its modulation by myriocin, which inhibits sphingolipids synthesis. The blended use of both targeted and untargeted mass spectrometry techniques unveiled an atypical lipid composition in broncho-epithelial cystic fibrosis cells and the derived extracellular vesicles. The airways cells in cystic fibrosis accumulate lipids, exacerbating inflammation that can be partly reversed by myriocin therapy, which acts directly on sphingolipid synthesis and indirectly on the accrual of other lipid classes. The CFTR dysfunction leads to an increased metabolism of sphingolipids, which in turn is connected to the release of ceramide-enriched extracellular vesicles that export a pro-inflammatory signal to the recipient cells. The accumulation of lipids may be the result of dysfunctional lipid traffic in the cell and may be responsible for blocking the ER-Golgi network, overproduction of reactive oxygen species and uncontrolled inflammatory stimuli. We proposed new insights on the role of lipid metabolism in the development of cystic fibrosis pathogenesis by using a validated mass spectrometry lipidomics approach. To conclude, we hypothesize that the metabolism of sphingolipids may be an effective pharmacological target to help the reduction of the intrinsic inflammatory state in the airways.
LIPIDOMICS UNVEILS THE ROLE OF BIOACTIVE LIPIDS IN CELLULAR MODELS OF CYSTIC FIBROSIS / M.v. Dei Cas ; coordinatore: M. SAMAJA ; tutor: M. SAMAJA ; cotutore: R. GHIDONI. Dipartimento di Scienze della Salute, 2021 Mar 17. 33. ciclo, Anno Accademico 2020. [10.13130/dei-cas-michele-vittorio_phd2021-03-17].
EVALUATION OF PHARMACEUTICAL AND COSMETIC USE OF EXCIPIENTS OBTAINED FROM CITRUS PROCESSING WASTE PRODUCTS
Depletion of natural resources and pollution are pushing society towards the development of models based on sustainability and recycling. In recent years, the use of food industry waste as new materials in pharmaceuticals and cosmetics has attracted increasing interest. In this circular economy perspective, bergamot peel waste is an established source of high-value components such as essential oils, flavonoids, pectin, and ascorbic acid. After the extraction of such phytochemicals, the remaining biomasses can be further used to recover residual lipid traces that have not been investigated so far. This doctoral thesis aimed to investigate whether bergamot lipid components (BLCs) remaining in the bergamot discarded biomass could be a functional excipient for the preparation of dosage forms for pharmaceutical or cosmetic purposes. Keeping in mind this aim, a suitable extraction method was developed and the composition of the obtained BLCs extract was determined by using Gas Chromatography with Mass Selection Detector (GC/MS) and ATR-FTIR spectroscopy. The raw extract was predominantly composed of long and medium-chain fatty acids with 70 % saturated and 30 % unsaturated acids. ATR-FTIR spectroscopy suggested that the extract appeared to be primarily composed of waxes and, to a lesser extent, free fatty acids. Since waxes are among the most used components—alongside triglycerides—in the preparation of solid lipid nanoparticles (SLNs), the possible application of BLCs as main components of SLNs was investigated. However, SLNs have limitations such as low drug encapsulation efficiency and poor long-term stability, often due to lipid polymorphic transitions. Therefore, in addition to formulating SLNs using only BLCs, the production of nanostructured lipid carriers (NLCs) by incorporating a liquid lipid, which can prevent the crystallization of primary lipid, was also investigated. The most promising formulations were finally evaluated for delivering solid or liquid drug through the skin and pre-gastric mucosae. For what concerns transdermal drug delivery, these nanosystems were found to be effective skin penetration enhancer due to their ability to induce a conformational change in stratum corneum lipids, shifting them from an ordered configuration to a liquid-like organization upon penetration of BLCs. The hypothesis of perturbation of the stratum corneum lipid network was further demonstrated by a second set of in vitro permeation studies performed by the co-administration of placebo BLCs nanosystems added to ibuprofen solutions or gels. The co-administration did not lead to significant differences compared to drug-loaded nanosystems but highlighted a significant increase in drug skin penetration compared to the corresponding dosage forms (solution and gel). Finally, the lack of interaction with keratins, as confirmed by the ATR spectra, suggested that the perturbation of the human stratum corneum occurred in a reversible manner, ensuring the BLCs safety. To definitively confirm that the mechanism of action of BLCs was exclusively based on the perturbation of the packing of the stratum corneum, the performance of BLCs nanosystems loaded with curcumin was also tested after application on another stratified epithelium, namely the oesophagus, using liposomes as references. As expected, the penetration of porcine oesophageal mucosa occurred be less efficient than the liposomal formulations. Besides clarifying the skin penetration effect of BLCs, the results also elucidated other technological aspects of BLCs related to the formulation of lipid nanosystems. Firstly, monodispersed nanoparticles can be easily obtained at relatively low temperatures (60-70°C) by a melting and emulsification method assisted by ultrasounds without the use of stabilizers, which are necessarily required in the formulation of SLNs and NLCs prepared with other excipients. Secondly, the peculiar composition of BLC
EVALUATION OF PHARMACEUTICAL AND COSMETIC USE OF EXCIPIENTS OBTAINED FROM CITRUS PROCESSING WASTE PRODUCTS / P. Volonte' ; tutor: P. Minghetti ; supervisore e coordinatore: G. Vistoli. Dipartimento di Scienze Farmaceutiche, 2025 May 13. 37. ciclo, Anno Accademico 2023/2024.
LA NOZIONE DI AIUTO DI STATO E LA NECESSITÀ DI UNA NUOVA MODERNIZZAZIONE DELLA DISCIPLINA
The purpose of this dissertation is to analyse the notion of State aid, with the aim of understanding whether it is still adequate to respond to the new needs inherent to the European single market as a result of recent economic and political events.The analysis starts from the examination of State aid competences, since they are the backbone of the whole system, which, as is well known, is regulated by Art. 107 et seq. TFEU. In this context, the focus is on the intersection between the primary authority of the Member States, which, at least at an initial stage, have full discretion as to the granting of a measure of advantage, and the strong powers of the European Commission, the only institution entrusted with aid control.Having examined how the intersection between these competences constitutes a point of encounter and, in some respects, of collision in this field, this research analyses the genesis and evolution of the notion of State aid, in order to understand the path that led to its current definition contained in a 2016 Commission notice (the “Commission Notice on the notion of State aid as referred to in Article 107(1) of the Treaty on the Functioning of the European Union”), which is also discussed in detail.Having established the foundation, the focus then shifts to the impact that two recent events have had on the State aid regulation: the Covid-19 pandemic and the Russian-Ukrainian conflict. The economic crises they generated resulted in State aid becoming one of the most widely used tools to provide relief for the real economy: in this context, the temporary frameworks issued to deal with the situation are analysed, as well as the interpretation that was given to the notion of aid.In the same perspective of understanding how modern events may have conditioned the definition in question (and vice versa), the paper first looks at the Brexit phenomenon and the difference between the European notion of “State aid” and the international notion of “subsidy”, and then inserts a third term of comparison given by the concept of “foreign subsidies”, which is the topic of a very recent protectionist regulation issued by the EU legislator.Finally, an extensive reflection is dedicated to the prospects of modernising the notion of State aid, where it is argued that the start of a new innovation process may be the key to the correct maintenance of level playing field.
LA NOZIONE DI AIUTO DI STATO E LA NECESSITÀ DI UNA NUOVA MODERNIZZAZIONE DELLA DISCIPLINA / E. Latorre ; tutor: F. Rossi Dal Pozzo; coordinatore: F. Biondi. Dipartimento di Diritto Pubblico Italiano e Sovranazionale, 2023 Jul 04. 35. ciclo, Anno Accademico 2022.
L¿INFLUENZA DELLE SENTENZE DELLA CORTE DI GIUSTIZIA PRIMA, DURANTE E DOPO IL PROCESSO AMMINISTRATIVO
Due to the significant resonance assumed in recent years by the increasing integration of European law into national legal systems and, above all, in light of the contribution that Luxembourg judges are more and more frequently asked to give in order to (re) define (or confirm) internal rules of procedure, the present study aimed to investigate the influence of the European Court of Justice’s judgements in the administrative trial. After outlining the historical and legal reasons that led Member States to cede a portion of their sovereignty to establish a sui generis legal system - the study focused on the major role played by the Court of Justice before the preliminary ruling procedure referred by the national court. Indeed, it was showed how this procedure is critical in the relationship between the EU system and national legal systems, being not only a means of control in the implementation of EU law, but also a guarantee of the subjective legal positions of individuals. The study then addressed the effects that EU jurisprudence has produced on the different stages of administrative judgment to focus on some of its fundamental institutions: after observing the influence of the Court of Justice’s judgements on the definition of European sources effects’ extent and in the resulting improved delimitation of any conflicts between national administrative measures and European law, the paper focused on legitimacy and appeal, also with reference to challengeable acts before the administrative judge. Moreover, the work examined the influence that the EU jurisprudence has had on precautionary and compensatory measures, as well as the consequences it has led to with respect to the internal procedural rules relating to the inviolability of the national res judicata. This influence was finally explored in its different aspects in terms of the actionable remedies to avoid the issuing of a judgment in conflict with European law and the configurability of an action of State responsibility, as a remedy to protect the injured party by a ruling of the administrative court of last instance issued in violation of the European Union law as interpreted by the Court of Justice.
L¿INFLUENZA DELLE SENTENZE DELLA CORTE DI GIUSTIZIA PRIMA, DURANTE E DOPO IL PROCESSO AMMINISTRATIVO / V. Sordi ; coordinatore: F. Biondi ; tutor: M. Sica. Università degli Studi di Milano, 2022 Jun 06. 34. ciclo, Anno Accademico 2021.
CELLULAR AND MOLECULAR MECHANISMS OF CHEMORESISTANCE IN ACUTE MYELOID LEUKEMIA
"Acute Myeloid Leukemia (AML) remains an incurable disease, with a 5-year survival rate below 30%. While the introduction of new therapeutic approaches, such as targeted therapies and immunotherapy, has had a modest impact on AML prognosis, the standard treatment of AML is still based on chemotherapy regimens (like ""7+3"") that have been in use for over 50 years. Approximately 20% of patients do not respond to standard chemotherapy, and even among those who initially respond, around 50% eventually relapse due to the development of chemotherapy resistance. Chemoresistance is the cause of death for over 90% of AML patients.The mechanisms underlying chemoresistance in AML are not yet fully understood, and targeted therapies aimed at preventing chemoresistance are not currently available. Chemoresistance in AML is primarily driven by non-genetic mechanisms. While leukemic stem cells (LSCs) have been implicated, recent studies suggest that AML blasts that survive chemotherapy, known as Chemotherapy Persistent Blasts, enter a senescence-like and diapause state. Though they can contribute to disease recurrence, these Persistent Blasts are not inherently therapy-resistant as they revert to a drug-sensitive state upon drug withdrawal. However, a lack of robust in vivo models of clinical chemoresistance in human AML hinders our understanding of these mechanisms. To address this, I aimed to generate in vivo models of truly chemoresistant AML to investigate non-genetic mechanisms of chemotherapy resistance.I utilized mouse models of human AML, specifically Patient-Derived Xenografts (PDXs), to establish treatment protocols mimicking the standard ""7+3"" chemotherapy regimen used clinically. One model (AML9) mirrored the behavior of most patients treated with standard chemotherapy, exhibiting clinical remission followed by relapse. In contrast, the other model (AML20) showed a modest response to chemotherapy with rapid leukemia re-expansion. Notably, relapsed leukemias in both models were completely resistant to retreatment with the same chemotherapy regimen, and this resistant phenotype persisted even after re-transplantation. Extensive genomic profiling revealed that chemoresistance in relapsed leukemias was not associated with the emergence of de novo DNA mutations. AML9 and AML20 represent the first two available AML models of true clinical chemoresistance, mirroring primary and secondary resistance, respectively.9To investigate mechanisms of chemoresistance in these models, I characterized the cellular and molecular properties of Chemotherapy Persistent Blasts isolated at the end of the chemotherapy regimen. To monitor and isolate quiescent blasts during leukemia growth and after chemotherapy treatment, I incorporated a label-retaining (LR) assay into the PDX models, enabling cell division tracking through inducible expression of H2B-GFP. I found that: i) in vitro, Persistent Blasts from both models exhibited significantly higher IC50 values compared to non-treated blasts, suggesting the induction or selection of a stable and cell- autonomous chemoresistant phenotype; ii) Quiescent blasts were slightly enriched among Persistent Blasts, but a portion of leukemia cells continued to proliferate during treatment, generating a population of Persistent Blasts of both proliferating and quiescent cells; iii) Persistent Blasts retained regenerative potential in vivo, primarily attributed to persistent quiescent blasts, suggesting that quiescence, while not a specific trait of chemoresistance, is integral to the relapsing properties of these cells; iv) the AML clonal structure remained unchanged after chemotherapy treatment, as determined by lineage tracing of Persistent Blasts and relapsed leukemias, suggesting that chemotherapy does not select specific cellular lineages; v) Persistent Blasts exhibited distinct transcriptional features compared to untreated leukemias, characterized by stem-cell signatures and activation of the Interferon ("
CELLULAR AND MOLECULAR MECHANISMS OF CHEMORESISTANCE IN ACUTE MYELOID LEUKEMIA / S. Frascolla ; tut: P. G. Pelicci ; phd coordinator: D. Pasini. - Milano. Università degli Studi di Milano, 2025 Jan 21. 36. ciclo
THE PLURALIST PHILOSOPHIES OF THE FRENCH AND ANGLO-AMERICAN TRADITION: A GENEALOGY OF ONTOLOGICAL PLURALISM
The aim of our thesis is to construct a genealogy of ontological pluralism in the Twentieth century French and Nineteenth and Twentieth century Anglo-American tradition. The reason which guides our thesis is the growing interest which these two traditions have elicited in some contemporary philosophical debates, especially among the philosophers of the New Materialisms movement. In fact, many new materialists have expressed a keen interest in the pluralism upheld by both the French and Anglo-American tradition and they have often made use of the historical intertwinement of these two schools of thought. Nonetheless, said intertwinement has been sorely understudied making it both a relevant and yet overlooked field of inquiry.Our thesis starts with a survey of the most relevant figures of the New Materialisms movement and their interest in the French and Anglo-American tradition. Secondly, it moves on to the actual genealogy itself, concentrating specifically on three key figures in the French tradition, whose reading of the Anglo-American tradition, we believed, shaped the way in which the question is approached in our contemporary debates: Henri Bergson, Jean Wahl and Gilles Deleuze. The three thinkers are analysed in order to probe their respective relationship with the Anglo-American tradition. Lastly, in the conclusion we tied the contemporary debates and the historical genealogy together, showing how the French reading of the Anglo-American tradition foreshadowed the current state of the New Materialisms debate.The end-goal of our thesis is, overall, to argue both for the relevance of these two traditions and to bring to light the untapped potentials they could still bring to our current philosophical debates. The main objective is, in other words, to analyse what has already been done by the New Materialists and, at the same time, re-actualize the problems which still linger at the heart of these two often overlooked schools of thought.
THE PLURALIST PHILOSOPHIES OF THE FRENCH AND ANGLO-AMERICAN TRADITION: A GENEALOGY OF ONTOLOGICAL PLURALISM / E. Monacelli ; relatrice: R. Fabbrichesi ; coordinatore: A. Pinotti. Dipartimento di Filosofia Piero Martinetti, 2023 Sep 22. 35. ciclo, Anno Accademico 2022.
This thesis dissertation addresses the question raised by the Anthropocene of rethinking theconcept of autonomy which is no longer viable in the conditions of the new epoch. It exploreshow nature can be considered a condition of autonomy understood as the self-realisation of theindividual identity. It also explores the notion of autonomy as self-limitation, that is, the ideathat freedom can be compatible with planetary limits. Finally, it addresses some justice-relatedaspects, namely, climate and intergenerational justice as crucial aspects of a human rights-basedconception of the idea of living well within planetary boundaries.
AUTONOMY AND THE ANTHROPOCENE / E. Arcos ; tutors: M. Ricciardi, L. Sacconi. Dipartimento di Scienze Giuridiche Cesare Beccaria, 2023. 35. ciclo, Anno Accademico 2022.
COMPARISON AND EVALUATION OF RICE (ORYZA SATIVA L.) CROPPING SYSTEMS FOR SUSTAINABLE AGRICULTURE
"In recent decades, the global agrifood system has increasingly faced variety of challenges, including climate change, soil degradation, loss of biodiversity, economic instability and the growing demand for food driven by population growth. Traditional agricultural practices adopted in rice (Oryza sativa L.) production, often characterised by intensive monoculture, heavy reliance on chemical inputs, and extensive resource use, have significantly contributed to these problems. The Paris Agreement aims to limit the global temperature increase to below 2°C above preindustrial levels, necessitating rapid decarbonisation across all economic sectors. The global food system alone accounts for nearly one-third of total global greenhouse gas (GHG) emissions and poses a risk to achieving the Paris Agreement’s targets (Crippa et al., 2021). In the European Union (EU), agriculture contributes 12 per cent of the total GHG emissions. It is a major driver of environmental issues such as biodiversity loss and eutrophication (Leip et al., 2015).In reply, the EU has introduced the European Green Deal (European Commission, 2019), a policy initiative that includes the Farm-to-Fork and Biodiversity strategies. Its key targets include achieving net-zero GHG emissions by 2050, reducing fertiliser inputs by 20 per cent, shifting 25 per cent of farmland to organic management, and planting three billion trees by 2030. However, the specific policy measures remain undefined and are left to individual Member States. Achieving these objectives requires a fundamental transformation in the production and consumption of agricultural goods.Agroecology (AE) represents an integrated approach that includes ecological and social principles into the design and management of food and agricultural systems (Barrios et al.,2020). Recognized as a potential practice to meet these goals under the common agricultural policy (CAP), AE is seen by scientists as a promising approach to addressing these challenges (Peeters et al., 2021). Local examples have demonstrated that AE can enhance various ecosystem services (Boeraeveet al., 2020). However, there is a lack of comprehensive evaluation to determine whether the outcomes observed at local levels are scalable. Moreover, research has yet to assess whether scaling AE to the territorial (i.e., EU) level might introduce new challenges, such as those arising from absolute thresholds in land use or nutrient availability from biological source.This thesis aims to apply the concepts of agroecology and sustainability to rice farming at various levels: the bibliographic level to determine what exists in the literature, the field level to test what was found in the bibliography, and finally, the farm level to evaluate the farming system in terms of sustainability and agroecology. For this, different research methods were used to retrieve data, information, and knowledge and to arrive at an integrated assessment of rice farming activities with the tools that the scientific field provides.All the chapters of the thesis are strictly connected. In fact, from the systematic review work, ""organic"" is the most recurring keyword in CO2 mitigation strategies. Additionally, among the techniques to be adopted for mitigation, ""organic fertilisation"" is among those that receive the highest scores. These very important results helped decide on the field trials. Specifically, for organic fertilisation, a consortium of nitrogen-fixing biostimulant bacteria was tested for two years as a more sustainable addition to traditional mineral fertilizations. The second field study concerns the world of organic rice, with seed replication and an in-depth review of allelopathic rice varieties, which, due to their potential ability to compete with significant weeds, could be a more sustainable varietal choice in the future. In the final chapter of this thesis, after analysing the sustainability of rice cul"
COMPARISON AND EVALUATION OF RICE (ORYZA SATIVA L.) CROPPING SYSTEMS FOR SUSTAINABLE AGRICULTURE / G. Savoini ; tutor: S. Bocchi; co-tutor: F. Fava; coordinatore: R. Pilu; revisori esterni :V. Vaglia, F. Vidotto. - Via Giovanni Celoria, 2, 20133 Milano MI (Facoltà di Scienze Agrarie e Alimentari). Dipartimento di Scienze e Politiche Ambientali, 2025 Feb 03. 37. ciclo, Anno Accademico 2023/2024.
MINIMALLY INVASIVE SURGERY TO FACILITATE MICRO-IMPLANT SUPPORTED MAXILLARY SKELETAL EXPANSION IN ADULT PATIENTS.
IntroductionThe aim of the present study is to evaluate the skeletal modifications induced by maxillary expansion supported by palatal micro-implants and localized osteotomies produced with minimally invasive surgical technique in young-adult and adult patients.In the present investigation, osteotomies had a lower extension than the ones used in conventional surgically assisted rapid palatal expansion (SARPE), to adopt a minimally invasive surgical technique that can be performed with greater comfort for the patient and less post-operative sequelae. More in detail, the localized osteotomies were executed only in the anterior part of the midpalatal suture and bilaterally at the basis of the zygomatic process of the maxilla without involvement of the piriform rim. These areas represent a great resistance to the lateral maxillary movement. Furthermore, they are of simple surgical access and don’t present important arterial plexuses.Patients requiring micro-implant supported maxillary expansion and/or SARPE routinely undergo a pre-treatment cone-beam computed tomography (CBCT) of the skull, to plan the surgical operation and, one month after treatment a secondary CBCT for a surgical and orthodontic control.CBCT is a low radiation tomography, extensively used in maxillofacial surgery and in dentistry. For the implementation of this study, that aims at evaluating the efficacy of micro-implant-supported maxillary expansion in combination with localized osteotomies, only radiologic exams that are anyway needed for the planning and post-treatment evaluation were used.AimThe aim of the present study is to evaluate the advantages introduced in the treatment of maxillary constriction by the therapy with micro-implant supported Maxillary Skeletal Expander (MSE) and localized osteotomies in young-adult and adult patients.The main objective is to evaluate the efficacy of the technique, by measuring the movement of skeletal landmarks in the midface, particularly on the maxillary and zygomatic bones, and on the lateral wall of the nose, by comparing the pre-treatment and post-treatment CBCT.For this particular technique, a new methodology for digital planning of position of MSE and miniscrews on patient CBCT was developed. Furthermore, incorporation of 2 additional miniscrews to the original MSE design, which conventionally features only 4 miniscrews, was developed with the aid of computer aided design – computer aided manufactured (CAD-CAM) technology.MethodsThe study presented the following steps:▪ Development of a digital planning methodology for positioning the miniscrews and MSE appliance on pre-treatment CBCT▪ Development of a CAD-CAM methodology for incorporating 2 additional miniscrews to the original MSE design with 4 miniscrews▪ Selection of patients with age above 17 years, without congenital craniofacial syndromes, who require intervention of maxillary skeletal expansion▪ Acquisition of initial CBCT with 17 x 13.5 cm field of view (FOV)▪ Intervention of maxillary expansion supported by palatal micro-implants and localized osteotomies executed with minimally invasive surgical technique▪ One month after treatment, acquisition of post-treatment CBCT with 17 x 13.5 cm FOV▪ Analysis of skeletal modifications in the midface (maxillary bone, sphenoid bone, zygomatic arch, nasal cavity, etc.), by comparing the pre- and post- treatment CBCT with a 3D software (OnDemand software by Cybermed)ResultsThe new methodology allowed the digital planning of MSE and miniscrews positioning on patients’ CBCTs, and the incorporation of two additional miniscrews to the original MSE design through CAD-CAM technology.In the clinical trial, a total of four patients had an average age of 27.6 years (range 22.1 – 39.9 years).MSE appliance was activated by an average of 6.0 mm and generated a parallel split of the midpalatal suture of 3.4 mm, 3.0 mm and 3.6 mm at anterio
MINIMALLY INVASIVE SURGERY TO FACILITATE MICRO-IMPLANT SUPPORTED MAXILLARY SKELETAL EXPANSION IN ADULT PATIENTS / D. Cantarella ; tutor: M. Del Fabbro; co-tutor: A. B. Giannì. Università degli Studi di Milano, 2022 Mar 16. 34. ciclo, Anno Accademico 2021.
DIFFERENTIAL MODULATION OF SMOOTH MUSCLE CELL PHENOTYPE BY NEXT-GENERATION NICOTINE PRODUCTS AND TRADITIONAL CIGARETTE
Cigarette smoke is one of the most important modifiable risk factors for cardiovascular diseases (CVDs) since smoking is responsible for 20% of deaths from coronary heart disease. Cigarette smoke contains 7.000 different components, many of which are known to be cytotoxic, mutagenic or carcinogenic. In the effort to reduce the harm associated with conventional cigarettes caused by the toxicants in the cigarette smoke produced by the burning of tobacco, alternative next generation nicotine products (NGPs, such as E-cigarettes (E-CIG) designed to deliver nicotine through an aerosol state), have been developed. Our research group has previously demonstrated that cigarette smoke condensate affects the plasticity of smooth muscle cells (SMCs). The aim of my project was to evaluate the effects of cigarette smoke aqueous extracts (AEs) on SMC phenotypic switch potential, a key step in the atherogenic process, and to get more information on the potentially toxic effects of the new NGPs. The in vitro studies in human aortic SMCs, showed that AEs, obtained from a traditional (TC) and E-CIG, stimulated the expression of extracellular matrix genes (ECM) and at the same time, E-CIG showed a potent and faster induction of SMC proliferation and migratory activity, while TC slowed down these processes compared to control and E-CIG. Moreover, we observed that the stimulatory effects exert by E-CIG are probably due to its nicotine content. In addition, α-BTX a potent and selective antagonist of alpha7 nicotinic acetylcholine receptor (α7nAChR) activation counteracted nicotine-induced cell migration and proliferation. Furthermore, the incubation with TC and E-CIG AEs also affected cell morphology, with the extension of lamellipodia, and increased F-actin and RAC1 levels. Interestingly, E-CIG showed a suppressive effect on inflammatory markers that was reversed by blocking α7nAChR with α-BTX. Overall, these results suggested that E-CIG promotes a phenotypic modulation in SMCs. In fact, E-CIG extract induces changes in cytoskeleton organization enhancing a proliferative and promigratory behavior. Moreover, we can speculate that these effects are mediated by nicotine through α7nAChR activation. On the contrary, TC increases ECM gene expression and induces changes in cell morphology but it might probably contain some other chemical compounds that overwhelmed the stimulatory effects exerted by nicotine on proliferation and migration activity. In addition, we demonstrated that TC is the most effective in promoting an inflammatory response by inducing the expression of several cytokines and matrix remodeling genes. In conclusion, these results demonstrated that E-CIG induce SMC phenotypic modulation by promoting SMC proliferation and migration activity and by inducing an alteration in cytoskeletal organization. E-CIG also exerts a negative modulation of the inflammatory pathway reducing cytokines expression. Further, the inhibitory effect is due to nicotine, one of the major components of cigarette smoke, through the activation of α7nAChR. TC is less effective in inducing a proper phenotypic modulation of SMCs while it is the most effective in increasing markers of inflammation and matrix-degrading metalloproteinases thus, promoting vascular inflammation and the atherogenic process.
DIFFERENTIAL MODULATION OF SMOOTH MUSCLE CELL PHENOTYPE BY NEXT-GENERATION NICOTINE PRODUCTS AND TRADITIONAL CIGARETTE / I. Damiani ; tutor: A. Corsini, ; co-tutor: S. Bellosta ; coordinator: G. D. Norata. Dipartimento di Scienze Farmacologiche e Biomolecolari, 2022 Apr 04. 34. ciclo, Anno Accademico 2021.
INVESTIGATION OF CELL-MICROENVIRONMENT INTERACTIONS BY ATOMIC FORCE MICROSCOPY TECHNIQUES
The role of forces is fundamental in a wide variety of biological processes, such as cell adhesion, migration, proliferation and differentiation. The ability of cells to perceive the nanotopographical features of the surrounding microenvironment (i.e. the extracellular matrix, ECM), called mechanotransduction, is mediated by specific trans-membrane proteins, called integrins, clustered together in Integrin Adhesion Complexes (IAC). Unraveling which mechanical and nanoscale morphological properties of the ECM determine the IAC composition and tune specific cellular response, is particularly challenging.Works have shown that biocompatible nanostructured thin films, grown by assembling zirconia nanoparticles (ns-ZrO2) on a substrate by Supersonic Cluster Beam Deposition technique, possess a morphological disorder on nanometer scale with structural features that mimic topographical properties of the ECM. Experiments performed with the neuron-like cell line PC12 demonstrated a link between the nanotopography of the ns-ZrO2 films and mechanotransductive events, which eventually foster neuronal differentiation.During my three years of PhD, I have developed novel approach based on Atomic Force Microscopy (AFM) to quantify cell sensing of nanotopographical features of the microenvironment, represented by ns-ZrO2 films reproducing the nanostructured surface of the ECM. Using custom ns-ZrO2–coated colloidal probes, we have carried out a quantitative analysis of adhesion strength and distribution of IACs by AFM-based adhesive force spectroscopy. We deposited ns-ZrO2 on custom AFM colloidal probes5. Bringing these nanostructured colloidal probes into contact with the body of living PC12 cells, it was possible to measure the strength, number and distribution of the IAC bonds by AFM force spectroscopy6, for different morphological properties of the interface. Furthermore, I used these functionalized probes to characterize the role of the surface pericellular layer, known as the Glycocalyx, in relation to the cell capability to react to external stimuli.Eventually, along with my personal research project, I had the opportunity to participate to two external collaboration with the Istituto Nazionale dei Tumori and with Istitute of Nuclear Physiscs from the Polish academy of Science. These collaborations aimed to exploit the knowledge in the mechanobiolgy field to study the mechanical implications of cells and tissues in cancer development and survival.
INVESTIGATION OF CELL-MICROENVIRONMENT INTERACTIONS BY ATOMIC FORCE MICROSCOPY TECHNIQUES / M. Chighizola ; director of the School: M.Paris; supervisor of the Thesis: A. Podestà. Dipartimento di Fisica Aldo Pontremoli, 2021 Feb 25. 33. ciclo, Anno Accademico 2020. [10.13130/chighizola-matteo_phd2021-02-25].
A COMPUTATIONAL PERSPECTIVE AT THE HISTONE MODIFICATION LANDSCAPE OF THE MOUSE LIVER DURING AGING
Aging, seen as the progressive physiological change in an organism that leads to the decline of biological functions and the organism’s ability to adapt to external and internal stress, is a complex process that is genetically and epigenetically determined and environmentally modulated. At the epigenetic level, aging is known to involve alterations of histone modifications which are associated with transcriptional changes occurring throughout the entire life span of an individual. Rather than studying these epigenetic variations between ”truly old” and young mice, we are particularly interested in clarifying how time as a primary factor affects the dynamics of histone modifications in young adults which are still in the healthy phase of their life span. To systematically approach this concept, we report the genome-wide mapping of mono- and tri-methylated lysine 4 (H3K4me1, H3K4me3), and acetylated and tri-methylated lysine 27 of histone H3 (H3K27ac, H3K27me3) in the livers of healthy 3, 6 and 12 months old C57BL/6 mice. The comparison of different age profiles of histone H3 marks revealed global redistribution of histone H3 modifications with time, in particular in intergenic regions and near transcription start sites, as well as altered correlation between the profiles of different histone modifications. Moreover, feeding mice with a 30% caloric restriction diet, a treatment known to retard aging, preserved a younger state of histone H3 in these genomic regions. Taken together, our data provide one of the most comprehensive descriptions of the dynamics of genome-wide histone H3 modifications over time and upon reduction of caloric intake, in mammals.
A COMPUTATIONAL PERSPECTIVE AT THE HISTONE MODIFICATION LANDSCAPE OF THE MOUSE LIVER DURING AGING / R. Hillje ; tutor: E. Colombo ; added supervisor: L. Luzi ; phd coordinator: P. G. Pelicci. Dipartimento di Oncologia ed Emato-Oncologia, 2020 Dec 11. 32. ciclo, Anno Accademico 2020. [10.13130/hillje-roman_phd2020-12-11].
TOWARDS PATIENT-SPECIFIC MODELS IN HIGH GRADE SEROUS OVARIAN CANCER (HGSOC): LINKING EPIGENETIC TRACING OF CELL OF ORIGIN WITH ACTIONABLE ORGANOID MODELS
Ovarian Cancer (OC) is a major cause of cancer-related mortality, due to the late-stage diagnosis and failure of surgery and chemotherapy to fully eradicate the disease, that is reflected in a high rate of tumor relapse after treatments. Patients with high-grade serous ovarian cancer (HGSOC), representing the largest majority of OC (~70%), have not experienced significant improvement in overall survival in last decades (1), pointing to the acute need to identify new predictive biomarkers and therapeutic targets for clinical settings.This unresolved emergency derives from our poor understanding of HGSOC biology combined with the recognized lack of suitable clinically relevant models for this disease, as the available models fail to relate molecular aberrations to clinical histories (2). Moreover, one major challenge that sets HGSOC apart from all other solid tumors, that has witnessed significant progress in recent years, is the persistent uncertainty about its cell of origin and the consequent lack of molecular signatures for the unequivocal assignment of specific samples to either an ovarian surface epithelium (OSE) or a fallopian tube fimbrial epithelium (FI) origin, the two candidate tissues still debated in literature (3, 4). Altogether, these aspects have hampered the dissection of pathogenic mechanisms and the identification of targets for improved clinical care of patients, driving us towards the embracement of new paradigms for effective advancements in the clinical setting. To overcome these issues, in the past few years, in the lab, we have devoted our efforts to the identification of the cell of origin of the tumor and the development of a clinical model allowing the study of this disease. Our applications aimed at brought a valid boost towards those precision oncology approaches that promises to transform cancer care by delivering personalized treatments tailored to the genetic and epigenetic specificities of patients’ tumors, whose remarkable heterogeneity characterizes even seemingly homogeneous histotypes.This level of complexity urged the need to develop and validate patient-derived cancer models that recapitulate to a meaningful extent the features of primary or metastatic tumors, both for efficient drug development and for guiding therapeutic strategies. Among new technologies, patient-derived organoid cultures are emerging as powerful models able to recapitulate in a meaningful extent the features of primary tumors (5, 6). Their value was proved both in guiding therapeutic decision for broadly curable tumors (7) and in gaining new insights for intractable tumors such as pancreatic adenocarcinoma (8). OC studies have started to benefit from these advanced models only recently (9–11), and a huge part of this PhD work regards the generation of organoids models from both HGSOC and normal samples of FI and OSE, in order to encompass all kind of tissues useful for the study of the pathology. Indeed, while tumor organoids represent an in vitro proxy of OC at an advanced stage, normal organoids from healthy tissues can be used as control but also as an indefinitely expandable cellular model in which investigate tumor alterations in the context of a normal tissue. This is really significant also in light of our recent publications on the cell of origin of HGSOC (12), where we showed the establishment of a method that is able to stratify HGSOC tumors through an approach based on methylomics and transcriptomics that: i) allows to distinguish OSE- from FI- derived tumors; ii) allows to identify for each tumor subtype the specific transcriptomic alterations that could drive the tumorigenic process; iii) reveals a prognostic value for the tissue of origin of this disease.Taken together, these two sets of information could help in define the molecular mechanisms governing the disease at an unprecedented resolution, setting a paradigm for harnessing patient-s
TOWARDS PATIENT-SPECIFIC MODELS IN HIGH GRADE SEROUS OVARIAN CANCER (HGSOC): LINKING EPIGENETIC TRACING OF CELL OF ORIGIN WITH ACTIONABLE ORGANOID MODELS / R. Luongo ; supervisor: G. Testa ; co-supervisor: P. Lo Riso ; internal advisor: K. Havas Cavalletti ; external advisor: R. Fodde ; phd coordinator: S. Minucci. Dipartimento di Oncologia ed Emato-Oncologia, 2021 Dec 13. 32. ciclo, Anno Accademico 2020.
STUDY OF THE PALEONTOLOGICAL RECORD OF THE BESANO FORMATION (MIDDLE TRIASSIC) AT ¿SASSO CALDO¿, VARESE, UNESCO WHL MONTE SAN GIORGIO
The Besano Formation consists of an alternation of laminated dolomitic banks and bituminous shales, and sparse cineritic tuffs that are dated as Late Anisian–Early Ladinian. It is one of the richest fossil-bearing formations from the Monte San Giorgio area; on the Italian side of Monte San Giorgio, the Sasso Caldo site is the one from which the greatest part of the Besano collection housed at the Museo di Storia Naturale di Milano is from. This Ph.D. thesis aims to the study of the Besano Formation macrofauna, through biochronostratigraphic zonation of the Sasso Caldo Site, revision of the large ichthyosaur Besanosaurus leptorhynchus), and study of the most important specimens, chosen for their preservation and rarity, but also to test the hypothesis of variations in the influence of open sea on the Besano basin.All the available ammonoids and bivalves from the Sasso Caldo site (Besano Formation), housed in the collections of the Museo di Storia Naturale di Milano, were examined and determined. The systematic study led to the recognition of 15 ammonoid taxa belonging to 10 genera, and five species belonging to the bivalve genus Daonella. The study of bed-by-bed collected specimens allowed the biochronostratigraphic classification of the Sasso Caldo section and the time-calibration of invertebrate and vertebrate bioevents. Results evidence that at Sasso Caldo site crops out almost the entire middle to upper Besano Formation. corresponding the Nevadites secedensis ammonoid zone. The trend of distribution of specimens reflects the establishment of an intraplatform basin with discontinuous open-marine influence in the middle Besano Formation, while the upper Besano Formation corresponds to a shallower subtidal restricted platform environment.Among the terrestrial taxa recovered at Sasso Caldo from the upper Besano Formation, a remarkably well-preserved fossil scorpion (BES SC 1973) is described in this thesis. This finding corroborates the hypothesized existence of a near shoreline during the deposition of the upper Besano Formation. BES SC 1973 is assigned to a new taxon gen. et sp. nov., included in the family Protobuthidae. This finding represents the first arachnid recorded from the Besano Formation, and the second genus attributed to the family Protobuthidae. This specimen is also the first reported Italian Mesozoic fossil scorpion.Regarding vertebrates, MSNM V927 and 928, a portion of the axial skeleton of a large diapsid, is attributed to Helveticosaurus zollingeri, a rare diapsid known only from the Besano Formation. This animal was recovered in association with the ammonoid Ticinites, at the base of the N. secedensis Zone, in coincidence with the establishment of the intraplatform basin of the middle Besano Formation. This specimen is the first record of skeletal remains and the second specimen assigned to the taxon in Italy. In this work the niche occupied by this animal in the Middle Triassic coastal ecosystems and its swimming style are also revised and discussed.MSNM V926, and SMNS 50010, respectively a portion of ribcage and an isolated partial forefin of a large ichthyosaur, were attributed to Cymbospondylus buchseri. MSNM V926 represents the first specimen attributed to this taxon and recovered on the Italian side of Monte San Giorgio. A great part of this thesis is dedicated to the revision of Besanosaurus leptorhynchus. The specimens studied and attributed to Besanosaurus leptorhynchus preserve a remarkably complete cranial and postcranial anatomy so that this taxon can be now accounted among the best-understood Middle Triassic ichthyosaur taxa. The revision of the skull morphology of this taxon clarified long-standing controversies regarding its cranial anatomy and the taxonomy of shastasaurids from Monte San Giorgio. The six specimens here described represent a potential ontogenetic series composed of an embryo (inside the maternal cavity of BES SC 999
STUDY OF THE PALEONTOLOGICAL RECORD OF THE BESANO FORMATION (MIDDLE TRIASSIC) AT ¿SASSO CALDO¿, VARESE, UNESCO WHL MONTE SAN GIORGIO / G. Bindellini ; tutor: M. Balini ; co-tutor: C. Dal Sasso ; coordinatore: M. I. Spalla. Dipartimento di Scienze della Terra Ardito Desio, 2022 May 06. 34. ciclo, Anno Accademico 2021.
TRANSLATIONAL APPROACH TO NEUROMUSCULAR FUNCTION AND ITS RECOVERY: EFFECT OF LIMBS, AGING, AND DISEASE.
Several factors can determine changes along the pathway from the brain to the muscles, and theinteraction of different processes during exercise can ultimately determine neuromuscular functionfatigability. Furthermore, the time course of neuromuscular function recovery after exercise isessential for planning repeated daily activities and sports. My PhD work aimed to compare the effectsof different fatiguing exercises, populations, and interventions on neuromuscular function fatigabilityand recovery.Daily-living and sporting activities have different physical requirements, and the fatigability ofdifferent muscle groups is of interest. As a result, comparisons of the fatigability of upper- and lowerlimbmuscles have been investigated, but there is a lack of information about muscle force loss andrecovery following dynamic multi-joint exercise involving elbow flexors and knee extensors. Theaim of the first study was to evaluate the magnitude and etiology of neuromuscular function changesin elbow flexors and knee extensors from dynamic multi-joint maximal incremental exercise. For thisstudy 10 young, healthy, and physically active men were enrolled. Each participant completed onefamiliarization session and two experimental sessions consisted of a maximal incremental exercise totask failure on either an arm-cranking ergometer or a cycle ergometer. Before, exactly 1 min and 20min after exercise cessation, neuromuscular function evaluation was conducted. The results showedthat both maximal voluntary isometric force loss and decreases in evoked forces were not differentbetween elbow flexors and knee extensors 1 min after task failure. However, 20 min after task failure,maximal voluntary isometric force, and low-frequency fatigue were greater in elbow flexors thanknee extensors. These results suggest that exercise-induced recovery is muscle-specific.The neuromuscular function is affected by age and the changes within the neuromuscular system areassociated with a decline in both muscle strength and mass. Since aging affects functionalperformance tasks such as stair climbing, ambulation, and rising time from a chair, affectingneuromuscular function, it is of interest to investigate muscle force and power fatigability.Sarcopenia, a highly prevalent disease in older than 65 years, exacerbates age-related changes in theneuromuscular system. The second study aimed to determine the mechanistic bases of the kneeextensor muscles fatigability in healthy and sarcopenic old men and women elicited by high-velocityconcentric contractions. Fifty-seven healthy old, 14 sarcopenic, and 21 young adults were recruited.Neuromuscular function assessment was performed before and immediately after an intermittentisotonic fatiguing exercise. Sarcopenia was associated with a reduction in knee extensor contractileproperties, isometric torque, and power. Exercise-induced loss of isometric torque was not differentbetween healthy and sarcopenic adults whereas power reduction resulted higher in sarcopenicIIIsuggesting that the age-related changes in fatigability are contraction-specific and exacerbated bysarcopenia. Since no differences were observed for voluntary activation, the greater power reductionduring exercise in sarcopenic adults may be due to intrinsic muscle impairment of contractile and/ormetabolic nature.Many research activities have been addressed in counteracting the muscle weakness observed in olderpeople. Since, aging seems to negatively affect plasma nitrate concentration [NO3-] and the potentialbenefits of dietary NO3- supplementation on skeletal muscle contractile function and muscle fatiguehave been recently suggested, the third study aimed to evaluate the effects of prolonged NO3-supplementation on contractile properties and fatiguability of ankle plantar-flexor muscles in elderlysubjects. Eleven elderly and 11 young men were recruited. Two main experimental sess
TRANSLATIONAL APPROACH TO NEUROMUSCULAR FUNCTION AND ITS RECOVERY: EFFECT OF LIMBS, AGING, AND DISEASE / M. Colosio ; supervisors: G. Vernillo, R. Codella, S. Porcelli; coordinatore: C. Sforza. Dipartimento di Scienze Biomediche per la Salute, 2023 Jul 14. 35. ciclo, Anno Accademico 2022.
NEW ADVANCES IN QUANTITATIVE RADIOLOGY: RADIOMICS IN NEURORADIOLOGY APPLIED TO PRIMARY BRAIN TUMORS USING A MACHINE LEARNING APPROACH
Arterial spin labelling (ASL) radiomics analysis to predict IDH mutation and MGMT methylation status in gliomas Fabio M. Doniselli1,2, Riccardo Pascuzzo1, Eleonora Bruno3, Domenico Aquino1, Mattia Verri, Alberto Redolfi, Valeria Cuccarini1, Marco Moscatelli1,2, Maria Grazia Bruzzone1, Luca Maria Sconfienza2,4 Abstract Objectives: To evaluate the strength and ability of radiomics features extracted from multiple tumor subregions on MR brain images to predict MGMT promoter (MGMT) methylation status and isocitrate dehydrogenase (IDH) mutation in glioma patients through a multiparametric MRI-based radiomics model, using arterial-spin labelling (ASL) perfusion imaging. Methods: Retrospective single-institution study in a cohort of 52 glioma patients. Radiomics-based models with a minimal set of relevant features and clinical parameters were built for MGMT methylation and IDH-mutation prediction from a training cohort (31 patients) and tested on an validation cohort (13 patients). Results: Feature selection methods (Boruta, RFE and LR-EL) identified age and 3 radiomics features for MGMT prediction and 3 features for IDH prediction. For IDH prediction, SVM classifier achieved average 96.8% accuracy and 0.929 AUC during the training phase, and 84.6% accuracy and 0.60 AUC on the test set. For MGMT methylation prediction, SVM classifier achieved average 67.7% accuracy and 0.765 AUC during the training phase, and 38.5% accuracy and 0.429 AUC on the test set. Conclusions: The classification model based on both demographic (age) and radiomic ASL perfusion characteristics had the best performance in predicting the IDH mutational status of gliomas. This result suggests that the proposed method has promising efficacy in predicting IDH mutational status. We have not obtained a sufficient result trying to correlate radiomics with the MGMT mutational pattern.
NEW ADVANCES IN QUANTITATIVE RADIOLOGY: RADIOMICS IN NEURORADIOLOGY APPLIED TO PRIMARY BRAIN TUMORS USING A MACHINE LEARNING APPROACH / F.m. Doniselli ; tutor: L.M. Sconfienza ; coordinatore: M. Del Fabbro. Dipartimento di Scienze Biomediche per la Salute, 2022 Jul 05. 34. ciclo, Anno Accademico 2021.
THE IMPACT OF TRIPLE STELLAR SYSTEMS ON PROTOSTELLAR DISC DYNAMICS
Theoretical challenges in modelling multiple stellar systems are mirrored by observational limitations, with multiplicity often concealed within disc cavities and hidden by surrounding material. Nevertheless, advancements in observational instruments are steadily revealing multiple stellar systems within discs, while astrometry is becoming more sensitive. The present capabilities offer us the opportunity to develop a comprehensive theory for disc and planet formation in multiple stellar systems and prepare our models for forthcoming observational data.Throughout this thesis, I underlined the significance of taking into account multiple stellar systems in models of stellar and planetary formation. Ignoring the influence of multiplicity restricts the range of configurations for which our theories hold. Despite the added complexity, multiple stellar systems provide a unique way for a more profound understanding of the overall stellar and planet formation processes. The broader parameter space and diverse mechanisms inherent to these systems yield an abundance of observables that probe various aspects of ongoing physical processes, from gas hydrodynamics to dust dynamics and evolution. Importantly, these observables are indicative of fundamental hydrodynamical properties governing stellar and planet formation across diverse systems, not just those with two or more stars.In particular, this thesis challenges the oversimplification of reducing the dynamics of multiple stellar systems with more than two stars to two-body system dynamics. It asserts the critical importance of accounting for the secular evolution of these systems and highlights additional phenomena inherent to multiple stellar systems that can elucidate observations, providing otherwise inaccessible insights in stellar formation physics.
THE IMPACT OF TRIPLE STELLAR SYSTEMS ON PROTOSTELLAR DISC DYNAMICS / S. Ceppi ; supervisori: G. Lodato, N. Cuello ; coordinatore: R. Vecchi. Dipartimento di Fisica Aldo Pontremoli, 2024 Feb 05. 36. ciclo, Anno Accademico 2022/2023.
NONLINEAR WAVES EXCITATION AND INTERACTION IN PURE ELECTRON PLASMAS
Nonneutral plasmas are excellent subjects for well controlled studies on basic physics problems and industrial research over a wide range of parameters. For the long-time confinement, merger and recombination of antimatter, the method of choice is some variant of the Penning-Malmberg trap, and many of the techniques for the manipulation of charged particles, such as cooling, compression, transfer, and ultimately a stable confinement in a quiescent state, are based on methods first developed by the nonneutral plasma community using electron plasmas. Another fascinating properties of nonneutral plasmas is the fluid analogy: in a cold, magnetized, nonneutral plasma, the 2D transverse dynamics equations are isomorphic to the Euler equations for an ideal 2D fluid. Hence, a pure electron plasma in a Penning-Malmberg trap evolves as an inviscid, incompressible bidimensional fluid.The study and control of the various waves and instabilities is of great interest in physics, and it is often intertwined with non-linearity and turbulence. Experiments aimed at unveiling hidden dependencies between parameters or improve the control over non-equilibrium, unstable configurations can lead to the discover of new phenomena, and a more thorough understanding of basic plasma physics, like the excitation and interaction of different plasma modes in pure electron plasmas, is likely to be a useful tool for a range of applications.In this thesis work, we present the results of the experiments made on two different Penning-Malmberg traps on the excitation and interaction of nonlinear waves in pure electron plasmas. On the Eltrap device, located at the University of Milan, Italy, we perform experiments on the excitation and control of high order diocotron modes via the rotating electric field technique. On the CamV device, located at the University of California, San Diego, we investigate the newly discovered phenomenon of TG waves splitting due to the interaction with a diocotron mode.
NONLINEAR WAVES EXCITATION AND INTERACTION IN PURE ELECTRON PLASMAS / N. Panzeri ; tutor: M. ROME' ; coordinatore: M. PARIS. Dipartimento di Fisica Aldo Pontremoli, 2021 Jan 27. 33. ciclo, Anno Accademico 2020. [10.13130/panzeri-nicola_phd2021-01-27].
IL VALORE DEL 'PRECEDENTE' NELLA GIURISPRUDENZA DEL GIUDICE DELL'UNIONE EUROPEA
The value of “precedent” in the case law of the Court of Justice of the European Union and the attempt to identify its characteristics within the European Union legal framework cannot be proceeded without the preemptive analysis of the legal reasoning and of the creative role of the judge. The role of the precedent in the legal reasoning and in the interpretation of the European Union law, as well as the unanimously accepted capacity of the judge to “create” law, are to be indented as the pivotal conditions allowing the development of a stare decisis system. The second part of the present thesis is focused on the peculiar features of the judicial precedent in the case law of the Court of Justice. In particular, we could notice, firstly, the absence of a clear definition and distinction between the concept of ratio decidendi and obiter dicta; secondly, we underline the need to overcome the classical distinction between persuasive precedent and binding precedent, in order to properly evaluate the scale of value that the precedent can take following the use and the function that is playing from time to time. Moreover, the analysis of the efficacy of the precedent will be confined, in this second part of the work, to the “self-precedent” of the Court, meaning to the obligation for the judge to follow his own precedent. The analysis will then highlight the role of precedent in case it is overruled or distinguished by the Court: practice that, ultimately, has to be conceived as a confirmation, as well as a recognition, of the precedent’s value. Furthermore, in spite of the absence of a doctrine of precedent in the European Union legal system, we recall two circumstances in which the judicial precedent assumes a de facto binding value. Firstly, we analyse the use of precedent in the decision-making activity of the judge, and then those norms of the rules of procedure that required the Court of Justice to decide by explicit reference to the previous judgment. Lastly, the analysis will be focus on the efficacy of the Court’s precedent with respect to the Tribunal is the evaluation of the impact of the vertical relationship between the two judges and the extension to which the Tribunal is bound by the Court’s as well as in the relationship between the Court and the national judge.
IL VALORE DEL 'PRECEDENTE' NELLA GIURISPRUDENZA DEL GIUDICE DELL'UNIONE EUROPEA / M. Postiglione ; tutor: M. Condinanzi ; coordinatore del corso di dottorato: D. U. Galetta. Università degli Studi di Milano, 2020 Nov 20. 32. ciclo, Anno Accademico 2019. [10.13130/postiglione-miriam_phd2020-11-20].
L'ENERGIA DEL COMMERCIO. IL RUOLO DEGLI SCAMBI DELLE FONTI D'ENERGIA NELLO SVILUPPO DELLA DDR
In its short life, the German Democratic Republic was one of the most powerful symbols and one of the hottest Cold War flashpoints of the European continent. Its existence was marked by its ability to manage and maintain a balance in its relations with the two blocs, while at the same time ensuring a degree of internal stability that would guarantee the permanence of the SED regime.Like all other states embedded in the increasingly globalized marketplace of the second half of the twentieth century, the East German state apparatus soon realized that one of the indispensable elements for consolidating its position was to strengthen its economic system, and that the foundation for its success was a sound energy security policy.The interest of contemporary historical research in the energy sector and the impact of the oil shocks of the last century at the national and international level is growing, driven also by the need to understand the profound historical evolution of the networks of dependencies in the current energy market. However, the development of the energy sector of the GDR in particular, the use of sources by the state, the rules and the markets of energy exchange, remain understudied to this day.Therefore, this paper, offers a new reconstruction of East Germany's development through the use of a perspective that adopts energy trade as a prism of observation, with a chronological focus on the development in the 1970s and 1980s, and specifically analyzes the policies, dynamics, negotiations and routes of buying and selling primary sources (lignite and oil) and secondary sources (petroleum products and electricity).First, the genesis of the GDR's economic, then industrial and energy systems is outlined. In fact, in order to fully understand its subsequent transformation it is not possible to ignore the years during which the entire economic policy of the new State was conceived and imposed: thus, the general situation from which it evolved and the main features on which the new energy structure was based are outlined and framed in the general historical context of the birth of the “border state”, the dynamics of the Cold War and the creation of the various international economic organizations within the blocs.It then analyzes the evolution of the energy system, considering the social and international context that the GDR had to deal with, up to the inevitable cooperation with West Germany before reunification. The main features of the economic and energy policies of the five-year plans, their adaptation to domestic needs, the economic and political goals to be achieved, and the upheavals in relations within Comecon and with other non-socialist countries are then highlighted. Some specific and original features of the use of some sources at the expense of others, the development of traditional and new trade routes aimed at ensuring a certain energy security - and, consequently, the economic stability and the proper functioning of the industrial sector - of the East German state are explored; the desired objectives and the concrete temporary and long-term results of the policies implemented are highlighted, as well as the perception that the government apparatus had of their importance in the general framework of state security.The study reveals a close correlation between energy and economic dynamics, which at times conflicted with each other, but at other times underscored the political and international needs of the GDR. What also emerges is an abiding interest on the part of the state's governmental and decision-making apparatus in the energy trade, the securing of which remained until the very end one of the primary objectives of the SED's economic maneuvers and the foundation on which the country's stability was believed to rest, and which was therefore attempted to be managed to the very end, regardless of political propaganda.
L'ENERGIA DEL COMMERCIO. IL RUOLO DEGLI SCAMBI DELLE FONTI D'ENERGIA NELLO SVILUPPO DELLA DDR / A.m. Scognamiglio ; tutor: G. M. I. Lami ; cotutor: M. Elli. Università degli Studi di Milano, 2024 Sep 10. 36. ciclo
SUSTAINABILITY AND INNOVATION OF CATALYTIC PROCESSES FOR THE SYNTHESIS AND TRANSFORMATION OF HETEROCYCLIC SYSTEMS OF BIOLOGICAL INTEREST
Functionalized heterocycles represent one of the most widespread structural motifs found in both molecules of natural origin and in synthetic compounds exhibiting diverse activities across biological, pharmacological, agrochemical fields, and as materials endowed with different properties. For these reasons, the demand for new, cleaner, and sustainable processes for the syntheses of heterocyclic compounds is continually increasing, as the known heterocyclic-forming reactions are often expensive, wasteful, and produce by-products. Among the modern synthetic methods, transition-metal catalysed reactions have emerged as a powerful and efficient tool for constructing a variety of heterocycle rings, resulting in shorter reaction times and higher yields when compared to other well-known methods reported in literature. On these bases, the project proposed herein studies and develops synthetic strategies for heterocycles, based on different transition-metals i.e. copper, platinum, ruthenium, alongside organocatalysis and metal-free reactions. The involved reactions aim to be innovative, leading to sustainable, green, and cost-efficient preparation of this important class of compounds.
SUSTAINABILITY AND INNOVATION OF CATALYTIC PROCESSES FOR THE SYNTHESIS AND TRANSFORMATION OF HETEROCYCLIC SYSTEMS OF BIOLOGICAL INTEREST / L. Molteni ; tutor: E.M. Beccalli ; coordinatore: G. Vistoli. Dipartimento di Scienze Farmaceutiche, 2024. 36. ciclo, Anno Accademico 2022/2023.
COMPREHENSIVE INVESTIGATION OF LIGHT NEUTRON-RICH NUCLEI TO TEST MODERN NUCLEAR THEORETICAL MODELS
Light nuclei, in particular the ones on the neutron-rich side of the valley of stability, represent a fertile ground to look for new experimental evidence, which can be exploited to benchmark theoretical approaches. In this Thesis we provide an experimental investigation of neutron-rich nuclei from boron to oxygen, employing state-of-the-art experimental techniques and detection systems. Novel information is collected and compared with the most advanced theoretical predictions currently available.The research activity described in this work has been developed on three levels, each associated to a different research line, that mirror the characteristic structure of a nucleus: bound states below the particle emission thresholds, near-threshold states and unbound resonances. In fact, the first line of research aims at the investigation of bound states in neutron-rich C, N and O isotopes, the second focuses on near-threshold resonances (11B as a specific case), and the third on stretched unbound states in the continuum of 13C, 14N and 16O. Altogether, a rather comprehensive body of experimental information will be discussed and confronted with some of the most advanced theoretical interpretations currently available for describing light atomic nuclei, such as ab initio approaches and Shell Model including the coupling to the continuum.
COMPREHENSIVE INVESTIGATION OF LIGHT NEUTRON-RICH NUCLEI TO TEST MODERN NUCLEAR THEORETICAL MODELS / S. Ziliani ; supervisor: S. Leoni ; co-supervisor: A. Bracco ; director of the school: M. Paris. Università degli Studi di Milano, 2021 Dec 13. 34. ciclo, Anno Accademico 2021. [10.13130/ziliani-sara_phd2021-12-13].
EN PITHO TEN KERAMEIAN. ANALISI TIPOLOGICA, TECNOLOGICA E FUNZIONALE DEL PITHOS NELLA PEUCEZIA PREROMANA: IL CASO DI JAZZO FORNASIELLO (GRAVINA IN PUGLIA - BA)
The cultural importance of the pithos, the large container for foodstuffs, is emphasized both in written sources and in iconographic documents where it appears in scenes of daily life, sacred rituals, as well as in relation to mythical episodes. However, the most significant evidence of its importance is provided by the study of material culture. For this reason, in the present study, the preferred area of investigation is pre-Roman Peucetia, a region with a significant agricultural vocation, where numerous pithoi are attested from different excavation sites.In this paper, we consider the case study of the large containers from the Peucetian site of Jazzo Fornasiello (Gravina in Puglia). This site has been excavated by a team from the University of Milan since 2009, under the scientific direction of Professors Marina Castoldi and Claudia Lambrugo. The large containers remained in use for centuries, sometimes passed down through generations, being dismissed only when the dwelling, understood as a social unit, was deconstructed and abandoned. The pithos, due to the technical difficulties of production, is a valuable artifact that is repaired for a long time rather than replaced, and due to its considerable size, it is often unmovable. It is also important to note that, for its function as a container for foodstuffs, this ceramic artifact quickly became a symbol of family wealth.In an attempt to accurately understand how the large containers of Jazzo Fornasiello were used, Professor Silvia Bruni and her team from the Department of Chemistry at the University of Milan conducted specific investigations on traces of organic substances. The results, particularly from the analysis of wine markers, demonstrated that alcoholic beverages were definitely stored inside the pithoi at Jazzo Fornasiello.All diagnostic fragments, found in the excavations of Jazzo Fornasiello until the 2019 campaign, were carefully recorded. Comparing the catalogued fragments with published findings from other Apulian and Enotrian excavations, it was possible to obtain a typological classification of the pithoi attested in the site.Within the current research, wide space was dedicated to archaeometric analyses, conducted in close collaboration with Professor Giacomo Eramo from the Department of Earth and Environmental Sciences at the University of Bari “Aldo Moro”. The analyses were performed on a large sample of vases in order to fully reconstruct the production chain underlying the making of the pithoi. The present research, which included a study of the large containers for foodstuffs from all perspectives, clearly highlighted the fundamental cultural significance that this ceramic form must have had within the Apulian communities, and within the agricultural-pastoral community settled at Jazzo Fornasiello between the 6thand 3rd centuries BCE, in particular. Indeed, as a symbol of family wealth, it becomes a vehicle for family memory and a tool used in domestic rituals associated with the celebration of ancestor worship.
EN PITHO TEN KERAMEIAN. ANALISI TIPOLOGICA, TECNOLOGICA E FUNZIONALE DEL PITHOS NELLA PEUCEZIA PREROMANA: IL CASO DI JAZZO FORNASIELLO (GRAVINA IN PUGLIA - BA) / A. Lojacono ; tutor: C. Lambrugo ; coordinatore: F. Slavazzi. Università degli Studi di Milano, 2024 Jun 19. 36. ciclo
HYDROGELS FOR GROWTH OF LIVING MICRO-ORGANISMS
In this work, I successfully achieved the goals of my PhD project on culturing diatoms, structural colored bacteria, and microalgae in different hydrogels, as well as co-culture bacterial and microalgal cells in the same hydrogel system. In our first project, we developed an innovative approach to study diatoms embedded in hydrogels with controlled conditions, resulting in a straightforward technique to investigate species-specific photo-regulation of benthic diatoms. This work opens an opportunity for further research to better understand how individual diatom species respond to different light conditions and the mechanisms that regulate their endogenic rhythms. In the second project, we have synthesized and characterized various hydrogels with different morphologies as matrices to grow three structural colored bacterial strains. By changing hydrogel compositions and studying their mechanical and physical properties, we successfully developed several methods for the bacteria to form bacterial biofilm with structural color, both in 2D and in 3D hydrogel matrix, which may contribute to further production of sustainable living pigmentations. The co-culture of microalgae and bacteria suggested the microalgal cells can contribute to bacterial growth thanks to the oxygen they produce from photosynthesis.In our last projects, we developed two advanced methods to culture microalgal cells. Microalgal growth in 3D granular hydrogels provides a possible solution for cell aggregation problem in bulk hydrogels. The granular hydrogels are made of fully degradable and highly biocompatible polymers with a facile synthesis method. Due to the interconnected microporous structure, microalgae are able to migrate and proliferate within the hydrogels, enabling faster cell growth, higher photosynthetic efficiency, and less self-shading effect. We also embedded nutrients-encapsulated silica nano-capsules into GelMA hydrogels and used them to culture microalgae, higher cell density and biomass production were achieved by using this culture method, suggesting a potential application to use silica nano-capsules to provide essential nutrients for microalgal cultivation. This work has provided a promising technique for harvesting renewable microalgal bio-resources and have potential applications in both laboratory and industry.
HYDROGELS FOR GROWTH OF LIVING MICRO-ORGANISMS / H. Kang ; tutor: L. De Cola ; phd coordinator: G. Vistoli. - Via Mangiagalli, 25, Milano. Dipartimento di Scienze Farmaceutiche, 2025 Jan 29. 37. ciclo, Anno Accademico 2023/2024. [10.17605/OSF.IO/WQDU6].
HETEROCYCLIC Β-AMINO ACIDS AS MOLECULAR TOOLS FOR PEPTIDOMIMETIC SYNTHESIS AND ORGANOCATALYSIS
β-Amino acid( AAs) are analogues of α-AAs with a difference: an additional carbon atom between the carboxylic and amine groups. β-AAs are commonly considered not natural amino acids, although there are some examples of this class in nature as secondary metabolites or as components of complex natural products.Being not natural AAs, when they are inserted in a peptide structure they can confer major proteolytic stability to the system compered to natural AAs. In fact, human peptidases do not recognize beta-amino acids and are not able to hydrolyze peptide bond. Moreover β-peptides and α/β hybrid peptides can induce a predictable and well-defined secondary structure, such as helices, strands and loops that may be residue-controlled and biologically active. During the PhD activity the synthesis of non-natural cyclic β-amino acids has been performed. Each of these has been used for different applications as described below:- Synthesis of non-natural 3-Arylmorpholino-β-amino Acid as a PPII Helix Inducer- Synthesis of electrospun fibers using model peptide containing morpholino-β-amino acid scaffolds- Synthesis of nucleobase morpholino β amino acids from ribonucleosides as molecular chimeras for the preparation of photoluminescent materials - Synthesis of non-natural amino acids with Morpholine core as organocatalysts for enantioselective Aldol-condensation reactions - Synthesis of non-natural amino acid with isoxazoline core as β-turn inducer for the preparation of peptidomimetics - Photochemistry cyclization peptide. This part of my thesis was performed during my visiting period at University of Manchester (UK) in Leonori Group.
HETEROCYCLIC Β-AMINO ACIDS AS MOLECULAR TOOLS FOR PEPTIDOMIMETIC SYNTHESIS AND ORGANOCATALYSIS / F. Vaghi ; tutor: M. L. GELMI ; coordinator: G. ALDINI. Dipartimento di Scienze Farmaceutiche, 2022 Apr 04. 34. ciclo, Anno Accademico 2021.
ASSESSING ENDOCRINE ACTIVE SUBSTANCES¿ IMPACT ON THE IMMUNE SYSTEM THROUGH IN VITRO STUDIES
Endocrine active substances (EAS) are substances able to interact with the endocrine system and in case they lead to adverse effects on health they are called endocrine disruptors (EDs). They can be found mainly in the environment, food, cosmetics, pesticides, plastics, and industrial products. There is a close interplay between the endocrine and the immune systems and therefore EDs can putatively interfere with immunity. Over the past few decades, there has been a rise in the prevalence of specific diseases, notably immune-related conditions such as allergies and autoimmune disorders. Among the leading potential factors attributed to this increase, EDs have been suggested.The aim of this thesis project was to investigate the ability of well-known EDs or suspected EAS to modulate the human immune system. To date, we have put forth various in vitro tests that utilize both cell cultures and human-derived primary cells. These tests could serve the dual purpose of eliminating the need for animal experimentation and enabling a more thorough evaluation of chemical hazards before they enter the market.My thesis started with the use of glyphosate, a compound whose use is a subject of intense debate. Glyphosate is among the substances suspected of having endocrine active properties, although the European Food Safety Authority (EFSA) recently affirmed that glyphosate cannot be described as ED. It is one of the most used herbicides worldwide, which safety has been recently posed under discussion. Even if still under debate, several endocrine disturbances (i.e., reproductive and thyroid) of glyphosate have emerged and some evidence also suggests possible immunotoxic effects in vivo (i.e., lung inflammation, rhinitis, rheumatoid arthritis). Therefore, the ability of glyphosate to directly interfere with immune cells activation was investigated. To this end, peripheral blood mononuclear cells (PBMCs) obtained from healthy donors were used to assess the possible interference with the immune system, focusing mainly on T helper cells differentiation. A reduction of T helper 1 cells was the main effect observed following glyphosate exposure, which resulted in an unbalance in Th1/Th2 ratio, favoring Th2 which may support the in vivo data. This effect was mediated by the estrogen receptor-α and by the up-regulation of a miRNA (miR-500a-5p). Accordingly, glyphosate demonstrated the ability to directly act on T cells, with a mechanism suggesting the involvement of estrogen receptor-α.Subsequently, my research activities were carried out within the PRIN project number 2017MLC3NF – EDONIS, focusing on the assessment of the immunotoxic potential of other EAS. To this end, six well known EDs or EAS were selected, and a battery of in vitro test was applied to determine their ability to interfere with the immune system. Atrazine, cypermethrin, vinclozolin, ethynyl estradiol, diethylphthalate, and perfluorooctanesulfonic acid were tested. The first three are pesticides, ethynyl estradiol is a pharmaceutical and lastly two are industrial chemicals. Initially, the human monocytic cell line THP-1 was used to investigate the ability of the selected compounds to modulate RACK1 (receptor for activated C kinase 1) expression, a protein considered as a bridge between the immune and the endocrine system, as its expression is under steroid hormones control. All the compounds modulated RACK1 expression paralleled by changes of several inflammatory markers (namely IL-8, TNF-α, CD54, and CD86). These effects were then confirmed in human primary cells of both sexes (PBMCs). In addition, natural killer cells lytic activity and T cells differentiation were also investigated. Based on the results obtained, perfluorooctanesulfonic acid and diethylphthalate resulted to be immunosuppressive on most parameters, and by using pharmacological tools we could demonstrate that the immunotoxic effects were endocrine-mediated.
ASSESSING ENDOCRINE ACTIVE SUBSTANCES' IMPACT ON THE IMMUNE SYSTEM THROUGH IN VITRO STUDIES / A. Maddalon ; tutor: M. Marinovich ; co-tutor: E. Corsini ; coordinatore: D. Norata. Dipartimento di Scienze Farmacologiche e Biomolecolari, 2023. 36. ciclo, Anno Accademico 2023.
DESIGN OF NOVEL MATERIALS FOR PHOTO-THERMO-CATALYTIC SOLAR FUELS PRODUCTION
The global challenge of transitioning to sustainable energy sources underscores the critical need for innovative technologies that can harness solar energy for fuel production. Among various strategies, the generation of solar fuels, such as hydrogen (H2) through water splitting or alcohol reforming, and the conversion of carbon dioxide (CO2) into valuable chemicals, has emerged as a highly promising route. Photocatalysis, which uses light to drive chemical reactions, offers a sustainable approach, but it faces significant limitations, primarily due to the low efficiency of light absorption and conversion by conventional semiconductor catalysts with wide bandgaps.A novel approach to overcome these limitations involves integrating heat with photocatalysis, a process known as photo-thermo-catalysis (PTC). PTC advantage the combined effects of light and heat, enhancing reaction rates and improving overall solar energy utilization. This PhD project aimed to develop and optimize catalytic systems for PTC, focusing on methanol steam reforming (MSR) for hydrogen production and CO2 reduction, while also deepening the understanding of reaction mechanisms through advanced in-situ techniques.The first phase of the project involved designing and building a bench-scale experimental setup capable of simultaneously studying the effects of light and heat on catalytic processes. Titanium dioxide (TiO2)-based materials, a well-known photocatalyst, were chosen as the initial focus. These materials were tested for hydrogen production via MSR, with and without the addition of noble metal nanoparticles like platinum (Pt), which are known to enhance charge carrier separation and improve catalytic activity.The study revealed that TiO2, both in its pure form and decorated with Pt nanoparticles, exhibited significant photocatalytic activity at room temperature. However, the introduction of heat substantially boosted the hydrogen production rate. Notably, Pt/TiO2 demonstrated two distinct activity regions: at lower temperatures, photocatalysis dominated, while at higher temperatures, thermocatalytic activity became predominant, and light still contributed in increasing the H2 productivity and the modulating the selectivity of the process. Despite the enhanced hydrogen production, a key challenge remained: the undesired formation of carbon monoxide (CO) as a byproduct, which limits the H2 productivity in the process. To address this, cerium oxide (CeO2) was incorporated into the catalyst formulation to promote the Water Gas Shift (WGS) reaction, which converts CO and water into CO2 and additional H2. Various synthesis methods and compositions were explored, revealing that moderate amounts of CeO2 improved catalytic efficiency and selectivity towards CO2, especially at mid-range temperatures. Interestingly, mechanical mixing of TiO2-based catalysts with higher CeO2 content provided the best overall performance, significantly enhancing hydrogen yield and selectivity through effective WGS activity.In the second phase, research expanded to explore LaFeO3 (LFO) based materials, a class of perovskite oxides noted for their visible-light activity, stability, and cost-effectiveness. Doping LFO with copper (Cu) and forming heterojunctions with other semiconductors were investigated as strategies to improve charge carrier dynamics and catalytic efficiency.The LFO-based catalysts showed promising results in both CO2 reduction and methanol reforming. The formation of heterojunctions with TiO2 enhanced CO2 reduction efficiency, while Cu-doping significantly improved hydrogen production via methanol steam reforming. These materials benefited from the combined effects of light and heat, underscoring the potential of PTC to enhance reaction rates and selectivity. Moreover, it was observed that introducing structural defects through mechanochemical treatments, while increasing photothermal conversion, i.e., the conversion of light into heat.To gain a deeper
DESIGN OF NOVEL MATERIALS FOR PHOTO-THERMO-CATALYTIC SOLAR FUELS PRODUCTION / S. Livolsi ; tutor: G. L. Chiarello ; coordinator: L. Prati. Dipartimento di Chimica, 2025 Jul 14. 37. ciclo, Anno Accademico 2023/2024.
PRINCIPIO DI PRECAUZIONE E DIRITTO PENALE: FONTI, DOGMATICA E DIRITTO VIVENTE
The work reflects on the use of the precautionary principle in the Italian Criminal law system in order to investigate its use in the legislation, in the juridical doctrine and in the court-law. This latest viewpoint is particularly enhanced so to give an updated and complete reconstruction of how the Supreme Italian Court (s.c. Corte di cassazione) has used the precautionary principle especially in culpable crime cases.
PRINCIPIO DI PRECAUZIONE E DIRITTO PENALE: FONTI, DOGMATICA E DIRITTO VIVENTE / M. Nicolini ; coordinatore: F. Poggi ; tutor: G. L. Gatta ; co-tutor: S. Zirulia. - Milano. Dipartimento di Scienze Giuridiche Cesare Beccaria, 2025 Jun 16. 37. ciclo, Anno Accademico 2023/2024.
BROWN ROT DISEASE DEVELOPMENT IN PEACH(P. PERSICA L. BATSCH): FROM FUNGAL BIOLOGY TO HIGH-THROUGHPUT ON-FIELDPHENOTYPING
"Brown rot (BR) disease, caused by Monilinia spp., causes significant pre-and post-harvest losses in stone fruit production, especially in humid and warm temperatures. In this thesis, we tried to tackle the subject with three complementary approaches. First, the recent progress in BR resistance in peach fruit was reviewed. Then we highlighted best practices in phenotyping BR susceptibility/resistance procedures in field and in vitro. We concluded that the main factors contributing to disease development are Monilinia inocula availability, environmental conditions, cultivars, fruit stage and management practices.Secondly, we investigated the anti-fungal effect of some phenolics such as chlorogenic and ferulic acids and triterpenoids such as oleanolic, betulinic, and ursolic acids. Furthermore, fruit surface compound (FSC) extracts of peach fruit at two developmental stages on Monilinia fructicola and M. laxa characteristics during in vitro growth were studied. A new procedure for assaying anti-fungal activity of triterpenoids, which are notoriously difficult to assess in vitro because of their hydrophobicity, has been developed. Also, a follow-up of this study revealed that certain phenolics and triterpenoids showed modest anti-fungal activity while dramatically modulating M. fructicola gene expression. MfRGAE1 gene was overexpressed by chlorogenic and ferulic acids and MfCUT1 by betulinic acid at 4- and 7-days post-inoculation.The third objective was to investigate the genetic background responsible for disease resistance in peach by detecting Quantitative Trait Loci (QTL) and attempts to identify molecular markers for assisted selection (MAS) in peach. For this, three F2 progenies, derived from three selfied F1 selections obtained from ""Contender"" (C, resistant) × ""Elegant Lady"" (El, susceptible), were investigated for two seasons (2019 and 2021). The whole progeny was genotyped by Single-Primer Enriched Technology (SPET) and a recently developed 18K SNP array. The genome-wide QTL analysis showed intriguing areas relevant to disease resistance, mainly the QTLs on chromosomes 2 and 4, which may be candidates for future MAS applications. Several new QTLs were detected for other fruit quality traits, including maturity date, soluble solid content and fruit weight."
BROWN ROT DISEASE DEVELOPMENT IN PEACH(P. PERSICA L. BATSCH): FROM FUNGAL BIOLOGY TO HIGH-THROUGHPUT ON-FIELDPHENOTYPING / M.h.m. Mustafa ; supervisor: M. Cirilli ; co-supervisor: D. Bassi ; reviewers: B. Quilot-Turion, I. A. Pacheco. Università degli Studi di Milano, 2022 Apr 27. 34. ciclo, Anno Accademico 2021.
FROM DATA PRODUCTION TO SEARCH FOR GALACTIC COSMICRAY SOURCES WITH THE PIERRE AUGER OBSERVATORY
Astroparticle physics, at the crossroads of astrophysics and particle physics, investigates ultra-high-energy cosmic rays (UHECRs), the most energetic particles known in the Universe. Despite decades of research, their origins remain elusive. These particles are detected through extensive air showers, cascades of secondary particles produced when UHECRs interact with the Earth's atmosphere. However, their low flux, less than one particle per square kilometer per century, makes detection extremely challenging.This PhD thesis, titled From Data Production to Search for Galactic Cosmic Ray Sources with the Pierre Auger Observatory, delves into the entire process of UHECR detection, from data production to the search for potential Galactic sources. The research is conducted using the Pierre Auger Observatory in Argentina, the largest cosmic ray observatory in the world, which employs a hybrid detection system to record cosmic ray events. Central to this setup is the Surface Detector (SD), consisting of 1660 water-Cherenkov detectors spread over 3000 square kilometers. This detector continuously records data with a 100% duty cycle, providing crucial information on the energy, direction, and composition of cosmic rays.The thesis emphasizes the meticulous processes involved in data production, including performance monitoring, event selection, and techniques to ensure high-quality datasets for analysis. A key contribution is the development of a new method for correlating cosmic ray events with potential sources in our Galaxy, using data sets above and below 1 EeV to investigate different energy thresholds and distances. This targeted search for neutron sources places new upper limits on their flux from potential Galactic objects, offering valuable insights into the nature and origins of UHECRs.The work represents a significant step forward in cosmic ray research, enhancing our understanding of data production processes and advancing the search for galactic cosmic ray sources.
FROM DATA PRODUCTION TO SEARCH FOR GALACTIC COSMICRAY SOURCES WITH THE PIERRE AUGER OBSERVATORY / F.m. Mariani ; supervisore: L. Caccianiga ; director of the school: A. Mennella. Università degli Studi di Milano, 2024 Nov 05. 37. ciclo, Anno Accademico 2024.
RESPONSABILITÀ DA REATO DEGLI ENTI E RIPARAZIONE AMBIENTALE
Reparative measures and sanctions for legal entities are two key policy options for an effective response against environmental crimes. Italian environmental criminal law includes several reparative obligations or mitigations/exclusions of the penalty due to reparation activities. These legal provisions address just the criminal sanction for natural persons. However, a closer look at the remedial activities involved demonstrates that, commonly, the individual can comply with reparative obligations only by using the means of the corporation. Analyzing the interplays between the individual and corporate dimension of remedial activities for environmental crimes is the aim of this doctoral thesis, in order to identify flaws in the current legislation and to advocate the needing for rationally reforming the environmental reparation measures, involving primarily the corporation and fitting with rules and policy goals of the legislative decree 231/2001.
RESPONSABILITÀ DA REATO DEGLI ENTI E RIPARAZIONE AMBIENTALE / M. Mossa Verre ; tutor: C. Paliero ; coordinatore: C.R. Luzzati. Dipartimento di Scienze Giuridiche Cesare Beccaria, 2022 Jul 07. 34. ciclo, Anno Accademico 2021.
CLINICAL APPLICATION AND PROGNOSTIC VALUE OF COMPUTED TOMOGRAPHY EXAMINATION IN FOALS DIAGNOSED WITH HEMATOGENOUS OSTEOMYELITIS
Bacterial sepsis in foals is associated with high morbidity and mortality. Bacteremia often arises from localized infections, with septic arthritis and osteomyelitis serving as potential gateways for bacterial spread or as sequelae of bacteremia.Computed tomography (CT) is increasingly utilized for the diagnosis and preoperative planning of osteomyelitis in foals. However, the association between CT findings and clinical prognosis is lacking in the literature. The objectives of this thesis were to; Provide a detailed resource of descriptive data of foals diagnosed with osteomyelitis, to compare findings obtained by conventional imaging methods and CT, and to investigate correlations between clinical and imaging findings and survival. Inclusion criteria were; 1. Foals under 9 months of age, 2. Presumptive clinical diagnosis of osteomyelitis, 3. CT examination used for further evaluation.Fifty-six foals were included in the final analysis. Thirty-six foals survived to discharge (36/56, 64%) and 26 foals survived long term (26/50, 52%). Variables with a significant negative effect on survival to discharge included articular involvement (p=0.04, OR 0.06 95%CI 0.04-0.89) and long transition zone (p=0.03, OR 0.133 95%CI 0.23-0.78). Variables with a significant negative effect on long-term survival included lesions in multiple locations p=0.03, OR 0.05 95%CI 0.04-0.77.In 93% of cases, CT examination modified the diagnosis, revealing improved lesion localization (72%), lesion size (76%), or detecting additional lesions (45%)These findings underscore the critical role of CT in identifying and localizing complex lesions ultimately improving prognostic assessments.
CLINICAL APPLICATION AND PROGNOSTIC VALUE OF COMPUTED TOMOGRAPHY EXAMINATION IN FOALS DIAGNOSED WITH HEMATOGENOUS OSTEOMYELITIS / K.e.v. Gustafsson ; supervisor : D.D. Zani ; coordinator F. Ceciliani. Dipartimento di Medicina Veterinaria e Scienze Animali, 2025 Sep 02. 37. ciclo, Anno Accademico 2024/2025.
PARKINSON'S DISEASE: VALIDATION OF A NOVEL TARGET THROUGH THE DEVELOPMENT OF METHYLPHENIDATE ANALOGS AS POTENTIAL MODULATOR AND FLUORESCENT PROBES
Parkinson’s Disease (PD) is a neurological disorder characterized by the progressive degeneration of dopaminergic striatal neurons, which in turn leads to the onset of a typical set of motor and non-motor symptoms. α-synuclein is the most studied protein involved in the pathogenesis of PD and its accumulation, misfolding and aggregation are strongly correlated with dopaminergic neuronal derangement. Synapsin-III is a neuronal protein involved in trafficking, fusion and binding of dopaminergic presynaptic vesicles. A growing body of evidence implicates the cooperation between α-synuclein and Synapsin-III in the pathogenesis of the disease and opens up to the possibility of pharmacological modulation of their interaction as a novel therapeutic approach for the treatment of PD. In Chapter 1 of this work, the design and synthesis of methylphenidate analogs as possible modulators of the α-synuclein/Synapsin-III interaction is presented. The parent molecule has been systematically modified in order to evaluate the effect of structural changes on biological activity. The obtained compounds have been tested for their biological activity and toxicity profile with the use of specific assays. The ability of these compounds to promote the interaction between the two target proteins, as well as their ability to slow down or prevent the aggregation of α-synuclein has been evaluated. Interestingly, one of the analogs showed a promising biological activity and, possibly, cytoprotective effects in cell models of PD. The activity data for the analogs will support the characterization of the binding modes to Synapsin-III and, hopefully, the identification of the preferential binding pocket. Moreover, very low or absent cytotoxicity corroborate our interest in these analogs as possible novel antiparkinsonian agents. In Chapter 2, the design, synthesis and radioligand binding evaluation of a novel group of fluorescent small molecule ligand-based probes for imaging the norepinephrine transporter (NET) is presented. NET is a member of the monoamine transporters family and is implicated in PD as well as in depression, ADHD and substance abuse. The selective imaging of NET in live cells, in particular with the aid of super resolution microscopy techniques, would contribute in a better understanding of its function, localization and trafficking at a molecular level. Herein, we identified a functional design for a NET-selective high affinity probe and verified its binding capabilities at NET. The most promising candidate is at the moment under evaluation in live-cell microscopy experiments. The work presented in Chapter 3 applies the fluorescent probes chemistry and concepts shown in Chapter 2 to the design and synthesis of novel fluorescent probes based on a methylphenidate scaffold and represents the ideal connection between the two projects previously described. The synthesized probes are now under evaluation in FRET experiments, in order to characterize the binding of methylphenidate on Synapsin-III, which would make a crucial contribution to our understanding of the interaction of the compounds presented in Chapter 1 with their target.
PARKINSON'S DISEASE: VALIDATION OF A NOVEL TARGET THROUGH THE DEVELOPMENT OF METHYLPHENIDATE ANALOGS AS POTENTIAL MODULATOR AND FLUORESCENT PROBES / A. Casiraghi ; supervisor: E. Valoti. Università degli Studi di Milano, 2020 Jan 22. 32. ciclo, Anno Accademico 2019. [10.13130/casiraghi-andrea_phd2020-01-22].
THE ROLE OF ENDOTHELIAL CELLS IN VASCULAR CALCIFICATION
Background and Aims. Chronic kidney disease (CKD) is a prevalent condition that affects millions of people. Patients with CKD have an increased cardiovascular risk, mostly due to vascular calcification (VC). By now, it’s quite clear how, in vitro and in vivo, vascular smooth muscle cells (VSMCs) actively participate to VC. In contrast to the wide studies on VSMCs, the role of endothelial cells (ECs) has been poorly investigated. It’s known that ECs have a dynamic nature and following particular stimuli they can trans-differentiate and acquire several different phenotypes through the endothelial-to-mesenchymal transition (EndMT).Methods. Human aortic ECs (HAECs) were challenged, in an in vitro calcification model, with 2, 2.5, 3, 3.5 mM Pi for up to 7 days. Early morphological changes, gene expression and cell migration were evaluated. Afterwards calcium deposition, genes and protein expression and ultrastructural changes were analyzed.Results. High-Pi, at a concentration of 3 mM, is able to induce EC calcium deposition. Calcification is dependent on calcium influx without influencing Pit-1 expression. During the first days of high-Pi treatment ECs change their morphology from cobblestone to spindle shape acquiring motility, as demonstrated by the 14% increased closure in the scratch wound healing test. EndMT master genes such as SNAIL and TWIST resulted up-regulated at day 3 together with an increased protein expression of mesenchymal markers such as N-Cadherin, α-SMA, SM22α, FSP-1 and Fibronectin. Starting from day 5 high-Pi induces osteoblastic differentiation with RUNX2 and BMP2 mRNA up-regulation up to day 7, osteopontin increased protein expression and positive Alcian Blu staining. In calcified ECs mesenchymal and endothelial markers, such as α-SMA, SM22α, FSP-1 and Fibronectin or vWF progressively decrease. Finally, uremic serum is able to exacerbate in vitro calcification, compared to healthy serum characterized by an increase in both mesenchymal and osteoblastic master genes such as TWIST and RUNX2.Conclusions. Our data demonstrate that in vitro high-Pi induces the EndMT process and simil osteoblastic trans-differentiation suggesting a potential role of endothelium in VC. The relevancy of endothelial calcification is suggested by the calcification exacerbation induced by uremic compared to healthy sera.
THE ROLE OF ENDOTHELIAL CELLS IN VASCULAR CALCIFICATION / L. Artioli ; relatore: M. Cozzolino ; co-relatore: P. Ciceri ; coordinatore: C. Sforza. Dipartimento di Scienze della Salute, 2024. 36. ciclo, Anno Accademico 2022/2023.
RESUMMATION AND MACHINE LEARNING TECHNIQUES TOWARDS PRECISION PHENOMENOLOGY AT THE LHC
The present thesis focuses on three distinct yet complementary areas in QCD. The first area concerns the resummation of large logarithmic contributions appearing in transverse momentum distributions. In particular, we focus on the hadroproduction of colour singlet final states such as Higgs boson produced via gluon fusion and the production of a lepton-pair via Drell--Yan mechanism. We present phenomenological studies of a combined resummation formalism in which standard resummation of logarithms of Q/pT is supplemented with the resummation of logarithmic contributions at large x. In such a formalism, small-pT and threshold logarithms are resummed up to NNLL and NNLL* respectively. The second area concerns the construction of an approximate NNLO expression for the transverse momentum spectra of the Higgs boson production by exploiting the analytical structure of various resummed formulae in Mellin space. The approximation we construct relies on the combination of two types of resummations, namely threshold and high-energy (or small-x). Detailed phenomenological studies, both at the partonic and hadronic level, are presented. And finally, the third area concerns the a posteriori treatment of Parton Distribution Functions (PDFs), specifically the compression of Monte Carlo PDF replicas using techniques from deep generative models such as generative adversarial models (or GANs in short). We show that such a GAN-based methodology results in a compression methodology that is able to provide a compressed set with smaller number of replicas and a more adequate representation of the original probability distribution. The possibility of using this methodology to address the problem of finite size effects in PDF determination is also investigated.
RESUMMATION AND MACHINE LEARNING TECHNIQUES TOWARDS PRECISION PHENOMENOLOGY AT THE LHC / T.r. Rabemananjara ; supervisor: S. Forte ; coordinatore: M. Paris. Dipartimento di Fisica Aldo Pontremoli, 2021 Dec 20. 34. ciclo, Anno Accademico 2021. [10.13130/rabemananjara-tanjona-radonirina_phd2021-12-20].
MALNUTRITION AND PHYSICAL PERFORMANCE IN NURSING HOME RESIDENTS: RESULTS FROM THE INCUR STUDY
Objectives: to investigate the association between malnutrition and physical performance in nursing home residents.Design: cross sectional study.Setting and Participants: a total of 499 older people (73.1% women) enrolled in the Incidence of pNeumonia and related ConseqUences in nursing home Residents (INCUR) cohort study.Methods: the nutritional status was defined using the Mini Nutritional Assessment–Short Form (MNA-SF), whereas the physical performance was measured with the Short Physical Performance Battery (SPPB). Unadjusted and adjusted (for age, sex, education, Abbreviated Mental Test score and Charlson Comorbidy Index) linear regression analyses were used to assess the association between MNA-SF and SPPB (primary outcome). The MNA-SF items which seemed most strongly related to the SPPB (and its subtasks) were also explored. Finally, we explored the association of the MNA-SF with each subtest of the SPPB.Results: the mean age of the sample was 85.80 (standard deviation, SD 7.20). The mean SPPB score was 3.05 (SD 2.65) and the mean MNA-SF score 10.35 (SD 2.22). Higher scores of MNA-SF were associated with better physical performance. Food intake deficiency, mobility impairment, and recent psychological stress or acute disease were the items of the MNA-SF most strongly associated with the SPPB, even after adjustment for potential confounders. Among the three subtests of the SPPB, the MNA-SF showed the strongest and most consistent association with the gait speed. Conclusions and Implications: among nursing home residents, the MNA-SF and three of its sub-items (food intake deficiency, mobility impairment, and recent psychological stress) are significantly correlated with physical performance, independently of potential confounders. In particular, the association was evident for the gait speed subtask of the SPPB.
MALNUTRITION AND PHYSICAL PERFORMANCE IN NURSING HOME RESIDENTS: RESULTS FROM THE INCUR STUDY / S. Damanti ; tutor: M. Cesari ; coordinatore: L. Pinotti. Università degli Studi di Milano, 2020 Jul 13. 32. ciclo, Anno Accademico 2019. [10.13130/damanti-sarah_phd2020-07-13].
CANCER-DERIVED EXTRACELLULAR VESICLES FOR TARGETED DELIVERY OF EGFRVIII SIRNA TO NEOPLASTIC TISSUE, EVALUATION OF THE HOMING MECHANISM AND POTENTIAL THERAPEUTIC APPLICATIONS IN GLIOBLASTOMA.
Extracellular vesicles (EVs), lipid-bound particles (50–150 nm in diameter), serve as natural delivery systems. Our previous data indicated a heterologous and cross-species tumour tropism for cancer-derived EVs such as colorectal cancer patient-derived EVs (PDEVs) and cancer cell-derived EVs compared to healthy donor EVs and non-transformed cell-derived EVs. For the first part of my PhD project, we evaluated some hypotheses to uncover the tumour tropism mechanism. These hypotheses centered around the half-life of EVs in circulation, EV interaction with cancer cells and extracellular matrix (ECM) and EV cargo release into the recipient cells.We first showed the eliminated tumour tropism feature upon filtering EVs. We also showed that trypsin treatment reduces EV-uptake rate by the cells. These experiments intensified the importance of EV corona since both trypsinization and filtration could affect the EV corona.Then we compared the EV uptake rate by cancer cells, transformed macrophages (RAW246.7) and primary macrophages (BDMSCs). Although, the results did not show superior uptake of cancer-derived EVs by cancer cells and RAW264.7, we observed unique interactions with BDMSCs, including large aggregate formation and phagocytosis deficits. This might lead to a higher half-life of cancer-derived EVs in circulation leading to their tumour site accumulation. We also showed a significant higher affinity of colorectal cancer-derived EVs to laminin as one of the major ECM components. Our mass spectrometry analysis (Proteomics) revealed differential protein expression for colorectal cancer patient-derived EVs (PDEVs) vs healthy donor EVs. Among these proteins, aminopeptidase N (CD13) showed a higher expression in cancer-derived EVs, confirmed by immunoblot analysis. Since CD13 is a potential protein involved in higher binding affinity of cancer-derived EVs to ECM component such as laminin and fibronectin through binding to NGR motif (Aspargine-Glycine-Arginine), we introduced CD13 as a potential candidate for the tumour tropism of cancer-derived EVs.For the second part of my PhD thesis, we harnessed the tumour tropism of glioblastoma (GBM)-derived EVs for targeted delivery of EGFRvIII siRNA, addressing this oncogenic driver in GBM. Among different approaches for loading siRNA into EVs such as endogenous loading, passive loading, membrane permeabilization, sonication, electroporation and transfection methods; transfection with a cell-penetrating peptide showed effective knock-down of EGFRvIII expression in the U373vIII cells. In conclusion, while EGFRvIII-targeting siRNA-loaded EVs demonstrated functional activity in vitro and partial efficacy in vivo, overcoming barriers such as correct dosing of siRNA loaded EVs, tumour microenvironment resistance, heterogeneous signaling pathways, and delivery inefficiencies will be critical for future therapeutic success. Combination approaches incorporating the EGFRvIII targeting with existing treatments like temozolomide (TMZ) or radiation could enhance overall survival outcomes. Further studies are necessary to refine EV-based delivery strategies and optimize therapeutic potential.
CANCER-DERIVED EXTRACELLULAR VESICLES FOR TARGETED DELIVERY OF EGFRVIII SIRNA TO NEOPLASTIC TISSUE, EVALUATION OF THE HOMING MECHANISM AND POTENTIAL THERAPEUTIC APPLICATIONS IN GLIOBLASTOMA / F. Shojaeighahrizjani ; tutor: P. Ciana ; co-tutor: J. Rak ; coordinator: G. D. Norata. - Milan. Dipartimento di Scienze della Salute, 2025 Apr 11. 37. ciclo, Anno Accademico 2023/2024.
INVESTIGATION OF ANTIBIOFILM AND ANTIVIRULENCE ACTIVITY OF 5-FLUOROCYTOSINE IN ESCHERICHIA COLI
Both acute and chronic infections by ‘opportunistic’ bacterial pathogens resistant to antibiotics are on the rise and they have been defined as a ‘slow motion catastrophe’ by the World Health Organization. Escherichia coli is an important opportunistic pathogen, and one of the most frequent causes of Urinary Tract Infections (UTIs). Due to the lack of novel promising antibiotics, novel alternative therapeutics are in demand. In particular, antivirulence agents, that selectively “disarm” pathogenic bacteria rather than killing them, are considered as a promising strategy. Several antimetabolites and nucleobase/nucleoside analogues, such as 5-fluorocytosine and 5-fluorouracil, have been shown to possess antivirulence activity in Gram negative bacteria like Pseudomonas aeruginosa and Escherichia coli. In my PhD work, based on previous observation that pyrimidine nucleotide availability can affect the production of biofilm and virulence factors in P. aeruginosa and E. coli, I evaluated fluoropyrimidines 5-fluorocytosine (5-FC), 5-fluorouracil (5-FU), and 5-fluorouridine (5-FUrd) as modulators of biofilm determinants in E. coli. I found that 5-FC showed lower antimicrobial activity to the E. coli laboratory strain MG1655 than 5-FU and 5-FUrd. However, all three fluoropyrimidines caused inhibition of curli production and general reduction of biofilm adhesion in dose-dependent manner in MG1655. This result was further confirmed by the observation that fluoropyrimidines can downregulate transcription of the curli structural gene csgA, suggesting that these compounds inhibit biofilm formation in E. coli through modulation of curli fibres expression. Since literature data indicate that 5-FC is less toxic to humans and my results proved that it possesses potent antibiofilm activity compared to other fluoropyrimidines, I focused on this molecule in further experiments. I tried to elucidate the molecular mechanisms behind the antibiofilm activity of 5-FC in E. coli and observed that curli inhibition by 5-FC requires its conversion into corresponding fluoronucleotide 5-FUMP. This led me to investigate the possible perturbation of de novo pyrimidine biosynthesis by 5-FC. Indeed, exposure to 5-FC resulted in a ca. 2-fold reduction of UMP intracellular levels, while not affecting the ATP nucleotide pool, thus suggesting a specific effect on the UMP biosynthetic pathway. Consistently, expression of the de novo pyrimidine biosynthesis genes carA and pyrB was upregulated in E. coli upon 5-FC treatment. Molecular docking studies suggested that 5-FUMP can bind the allosteric domain of carbamoyl phosphate synthetase, the first enzyme in de novo pyrimidine biosynthesis pathway, similar to its native allosteric inhibitor UMP and possibly mimic UMP’s negative feedback mechanism affecting the pyrimidine nucleotide pool. Overall, these results suggest that the antibiofilm activity of fluoropyrimidines is mediated, at least in part, by perturbation of the pyrimidine nucleotide pool. By performing E. coli MG1655 genomic library screening to identify additional determinants that counteracts the antivirulence effect of 5-FC, I identified a DNA fragment containing N-terminal domain of the penicillin-binding protein 1b (PBP1b), involved in peptidoglycan synthesis, and D8B36_18480, an unknown protein, restored curli production despite presence of 5-FC. Deleting PBP1b-encoding gene mrcB in E. coli increased curli gene expression, while overexpressing D8B36_18480 blocked it. While these two proteins do not appear to be direct targets of 5-FC in E. coli, they are potentially part of a not yet identified regulatory pathway linking peptidoglycan biosynthesis and curli production.Interestingly, along with exhibiting promising antibiofilm effect, 5-FC inhibit the expression of all major virulence factors of uropathogenic E. coli (UPEC) like curli fibers, secreted toxins, type I and P fimbriae, suggesting that the e
INVESTIGATION OF ANTIBIOFILM AND ANTIVIRULENCE ACTIVITY OF 5-FLUOROCYTOSINE IN ESCHERICHIA COLI / S. Ravishankar ; tutor : P. Landini ; cosupervisor/coordinator : E. Rossi. Dipartimento di Bioscienze, 2024 Nov 27. 37. ciclo, Anno Accademico 2024.
EXPLORING PHYSIOLOGICAL, PATHOLOGICAL AND PARAPHYSIOLOGICAL BRAIN ACTIVITY ACROSS SCALES WITH SIMULTANEOUS INVASIVE AND NON-INVASIVE EEG
"This thesis explores the integration of non-invasive high-density electroencephalography (hd-EEG) with invasive stereoelectroencephalography (SEEG) to enhance understanding of brain electrophysiology in pathological, physiological and paraphysiological contexts. The combination of these two methods, known as ""co-registration,"" leverages the high temporal resolution and full-brain coverage of hd-EEG with the spatial precision of SEEG, allowing for a more comprehensive view of brain activity. The thesis is divided into three primary areas of investigation.First, it examines pathological brain activity, specifically focusing on interictal epileptiform discharges (IEDs). These discharges, often difficult to detect with scalp EEG alone, were successfully localized using co-registration. By guiding hd-EEG analysis with SEEG data, hidden epileptic activity was revealed, contributing to more accurate non-invasive identification of epileptic foci, which is critical for surgical planning in drug-resistant epilepsy cases.Second, the study investigates physiological brain responses to sensory stimuli using median nerve stimulation. Co-registration enabled the analysis of how brain signals propagate from primary sensory regions to higher-order areas involved in perception. While hd-EEG captured fast, large-scale activity, SEEG provided detailed recordings from deep cortical structures. This integration revealed both phasic and tonic gamma band brain responses, offering new insights into sensory processing and conscious perception.Third, the thesis evaluates brain responses to artificial stimulation via single-pulse electrical stimulation (SPES), which evoked cortico-cortical evoked potentials (CCEPs). Co-registration allowed for a detailed analysis of how artificial stimulation propagates through brain circuits. While SEEG detected localized responses, hd-EEG captured broader patterns of brain activation, enhancing the understanding of brain connectivity and reactivity to external stimuli.Overall, the combination of hd-EEG and SEEG offers unique advantages, providing a more accurate and detailed picture of brain dynamics. This method not only refines neuroimaging techniques but also holds promise for improving clinical diagnostics and treatments, particularly in epilepsy. In essence, this thesis demonstrates that co-registration can bridge the gap between non-invasive and invasive brain recordings, offering significant benefits for both neuroscience research and clinical applications."
"EXPLORING PHYSIOLOGICAL, PATHOLOGICAL AND PARAPHYSIOLOGICAL BRAIN ACTIVITY ACROSS SCALES WITH SIMULTANEOUS INVASIVE AND NON-INVASIVE EEG / F.m. Zauli ; tutor: M. Massimini ; supervisori: A. Pigorini, E. Mikulan ; PhD coordinator: F. Guala. - DIPARTIMENTO DI SCIENZE BIOMEDICHE E CLINICHE ""LUIGI SACCO"" (MILANO). Dipartimento di Scienze Biomediche e Cliniche, 2025. 37. ciclo, Anno Accademico 2023/2024."
Obesity is a multifaceted condition often associated with poor outcomes in cancer, yet clinical evidence suggests that obese patients may benefit disproportionately from immunotherapy respect to the lean population. The mechanisms underlying this phenomenon, particularly in relation to sex, remain unclear.In this thesis, I investigated the impact of obesity and sex on tumor progression and response to anti–PD-1 therapy. Using the MC38 murine colorectal cancer model, we observed that high-fat diet (HFD) feeding accelerated tumor growth in both sexes, but more prominently in males. Nevertheless, the therapeutic benefit of anti–PD-1 was greatest in obese males, whose tumors exhibited markedly reduced progression compared to lean counterparts. These results align with clinical trends indicating improved outcomes in obese male patients, while highlighting sexually dimorphic effects of obesity on tumor immunity.To strengthen the robustness of our preclinical platform, we optimized the in vivo tumor model and applied a novel computational anti-clustering algorithm for animal randomization, which reduced baseline imbalances and enhanced reproducibility. My results revealed that macrophages, rather than CD8⁺ T cells alone, were the most consistently remodeled population in the tumor microenvironment, showing sex- and obesity-dependent infiltration. In vitro studies further demonstrated that lipid excess and estrogen differentially shaped macrophage inflammatory and antigen-presenting functions across sexes, with IL-6 emerging as a candidate mediator linking metabolic stress and immune regulation.Our findings provide mechanistic insight into how sex and obesity interact to modulate cancer immunotherapy, highlighting macrophages as central regulators of these effects, and underscore the importance of considering host-intrinsic factors in both preclinical modeling and the design of personalized immunotherapeutic strategies.
OBESITY AND SEX IN CANCER IMMUNOTHERAPY / F. Vicini ; tutor: L. Mazzarella ;co-tutor: P.G. Pelicci ;coordinator: D. Pasini. Università degli Studi di Milano, 2025 Oct 09. 37. ciclo, Anno Accademico 2024/2025.
GLI EPIGRAMMI 'DI PLATONE'. EDIZIONE, COMMENTO, FORTUNA
This thesis provides a new critical edition and full commentary of the epigrams transmitted under Plato's name. The edition includes for the first time all the epigrams assigned to 'Plato' by the ancient tradition and offers a critical text based on a review of the manuscript tradition and philological work. The commentary provides an in-depth analysis of the epigrams, from both the philological and the literary and exegetical points of view. An introductory study addresses the problem of the epigrams' authenticity by reviewing the scholarly debate on the topic. It also traces the history of Plato's epigrams in antiquity, proposing a reconstruction of relevant context and possible reasons why the epigrams were composed and assigned to Plato.
GLI EPIGRAMMI 'DI PLATONE'. EDIZIONE, COMMENTO, FORTUNA / M. Pelucchi ; tutor: G. Benedetto ; coordinatore: F. Slavazzi. Dipartimento di Studi Letterari, Filologici e Linguistici, 2022 Jul 08. 34. ciclo
INVESTIGATING GENOME PROTECTION AT G-QUADRUPLEXES: STRUCTURAL-FUNCTIONAL ANALYSIS OF S. CEREVISIAE DOMESTICATED TRANSPOSASE VID22
Genome instability is a characteristic feature of ageing and several human pathologies, including cancer. Genome instability is elicited by exogenous DNA damaging agents and by multiple endogenous factors, among which G-quadruplex non-canonical DNA secondary structures. These structurally heterogeneous nucleic acid folds have been implicated in the modulation of multiple biological processes, including transcription and replication. Consequently, perturbation of G-quadruplex homeostasis is linked to alterations in gene expression, replication stress, DNA breakage, and, ultimately, enhanced genome instability. We recently found that Saccharomyces cerevisiae Vid22, a putative member of the hAT family of DNA transposases by prediction of a BED-type zinc finger domain and an RNase H fold, binds at G-quadruplex forming sites and counteracts their detrimental effects on genome stability. In this work, I mutated various residues of Vid22 protein or the DNA sequence of one of its binding loci in the genome to investigate the intrinsic and extrinsic requirements for its recruitment to chromatin. Vid22 enrichment at certain loci depends on the presence of a G-quadruplex and contributes to the binding of Tbf1, a multifunctional general regulatory factor, to DNA. Moreover, Vid22 role in genome protection correlates with its physical interaction with Tbf1. Disruption of the putative DNA-binding BED domain only partially affects Vid22 interaction with DNA and Tbf1, whereas mutation of C-terminal residues abolishes both processes and correlates with cell hypersensitivity to genotoxic stress. Moreover, Vid22 forms homodimers in vivo. The computational model and low-resolution structure of Vid22 dimer display striking similarities to that formed by the active hAT Hermes transposase dimer despite their evolutionary distance and sequence divergence. Similarly to Hermes, Vid22 C-terminus seems to be crucial in maintaining the proper protein tertiary structure. Mutation of the C-terminus also abolishes Vid22 self-interaction due to its spatial proximity to a second putative DNA-binding region directly involved in dimerisation (residues 170-210) inferred by homology to Hermes, which existence is corroborated by the impairment of self-interaction in the vid22 F195A mutant. Thus, Vid22 has evolved from the domestication of a hAT DNA transposase to support genome stability and G-quadruplex regulation in budding yeast. Because the human genome also encodes uncharacterised proteins predicted to possess similar domain organisation and evolutionary origin, research on Vid22 may pave the way for their study, besides providing novel insights into G-quadruplex recognition and biology.
INVESTIGATING GENOME PROTECTION AT G-QUADRUPLEXES: STRUCTURAL-FUNCTIONAL ANALYSIS OF S. CEREVISIAE DOMESTICATED TRANSPOSASE VID22 / G.m. Bernini ; supervisor: M. Muzi Falconi ; tutor: F. Lazzaro ; coordinator: R. Mantovani. - Dipartimento di Bioscienze. Università degli Studi di Milano, 2024 Jun 25. 36. ciclo, Anno Accademico 2023.
LONG-LASTING MOLECULAR ALTERATIONS FOLLOWING EARLY LIFE STRESS IN RATS: ROLE OF GUT MICROBIOTA AND INFLAMMATION
It is well known that stress, especially when experienced early in life, affects the brain developmental trajectories leading to an enhanced vulnerability for several mental disorders later in life. The adverse outcomes depend on the timing of the stressful experiences, and in this contest, early stages, such as pregnancy and adolescence, represent key sensitive periods for brain development. Several studies in animal models have been carried out to understand the mechanisms underlying the negative outcomes associated with early adverse exposures. The presence of a chronic low-grade inflammatory status has been suggested to play a central role, however, a better knowledge of the inflammatory processes, as well as of the neurodevelopmental temporal window where such alterations become manifested, is still needed. Moreover, among the biological systems involved, the gut microbiota has recently been proposed to act at the interplay between the stress response system, brain functionality and inflammatory processes, through the gut-brain-immune axis communication. To better elucidate the involvement of inflammation and the gut microbiota in the long-lasting functional effects of stress exposure, we have used the prenatal stress (PNS) paradigm, a well-established preclinical model of stress during pregnancy that causes impairments in self-care, motivation and sociability and we have investigated possible long-lasting alterations in the inflammatory mechanisms. Specifically, we found that adult animals exposed to PNS show a significant cortical activation status of the p38 MAPK, NF-kB, Neuroinflammation and Dendritic cell maturation signalling pathways. We also found that PNS causes the most pronounced alterations in inflammatory mechanisms already at adolescence, highlighting the biological vulnerability of this temporal window. As we reported adolescence as the temporal window where biological vulnerability begins to manifest, we wanted to test whether pharmacological interventions during adolescence could modulate the impact of stress early in life. Therefore, we treated adolescent animals exposed to PNS with lurasidone, and we analysed their whole genome transcriptome. Our analyses indicated that none of the pathways related to pro-inflammatory processes were present in PNS animals treated with lurasidone, whereas we reported the implication of several processes related to inositol metabolism among the top involved mechanisms. As we also aimed at investigating the role of the gut microbiota, we tested changes induced by PNS exposure in the gut microbiota composition during different temporal windows both in male and female offspring. We reported that the gut microbiota significantly evolves during ageing both in female and male rats and that restraint stress during the last week of gestation produced several short- and long-lasting changes in the gut microbiota composition with a significant modulation of bacterial taxa such as Parabacteroides, Lachnoclostridium, Eubacterium ventriosum, Eisenbergiella, Turicibacter, Lactobacillus, and Muribaculum.As we observed that stress exposure during the prenatal period affects behaviour and it is associated with alterations in inflammatory mechanisms and in gut microbiota composition, we then wanted to evaluate the consequence of stress during a different temporal window (e.g., adolescence) on the same biological outcomes. Therefore, we investigated the effects of stress exposure during adolescence, by applying a social isolation paradigm, on central inflammation and gut microbiota. 16S sequencing analysis revealed that microbial changes were influenced by age in both isolated and control rats, regardless of sex, whereas social isolation impacted the microbial composition in a sex-dependent manner. Specifically, we found that social isolation induced short-term gut microbiota alterations in females but not in males. We also identified several stress-related bacterial taxa such as O
LONG-LASTING MOLECULAR ALTERATIONS FOLLOWING EARLY LIFE STRESS IN RATS: ROLE OF GUT MICROBIOTA AND INFLAMMATION / N. Lopizzo ; tutor: M.A. Riva ; co-tutor: A. Cattaneo ; coordinatore corso di dottorato: G. D. Norata. Dipartimento di Scienze Farmacologiche e Biomolecolari, 2022 Dec 12. 35. ciclo, Anno Accademico 2022.
The derived de Rham complex has been introduced by Illusie in 1972 as a natural consequence of the definition of the cotangent complex for a scheme morphism. This theory seems to have been forgot until the recents works by Bhatt and Beilinson, who gave several applications, in particular in $p$-adic Hodge Theory. On the other hand, the derived de Rham cohomology has a crucial role in a conjecture by Flach-Morin about special values of zeta functions for arithmetic schemes. The aim of this thesis is to study and compute the Hodge completed derived de Rham complex in some cases.
ON DERIVED DE RHAM COHOMOLOGY / D.m.a. Marangoni ; supervisors: B. Morin (Université de Bordeaux), N. Mazzari (Università degli Studi di Padova) ; coordinator: V. Mastropietro. Università degli Studi di Milano, 2020 Jul 28. 32. ciclo, Anno Accademico 2019. [10.13130/marangoni-davide-maria-alfonso_phd2020-07-28].
BLACK HOLES AND SOLUTION GENERATING TECHNIQUES
Multi-black hole solutions play a relevant role both from the theoretical and the phenomenological point of view. In this Thesis, we construct some regular multi-black hole spacetimes in pure Einstein’s General Relativity with the aid of solution generating techniques.We begin with a perspective on the history of solution generating techniques, and then we explain in detail the Ernst formalism and the inverse scattering method. These are the techniques that are applied in the rest of the Thesis.Subsequently, we construct multi-black hole solutions embedded in an external gravitational field: it is possible to obtain an equilibrium configuration in many interesting cases, like a collection of collinear static black holes or a chain of accelerating black holes, by choosing appropriately the multipole parameters of the field.Then, we consider the expanding bubbles of nothing as a background for multi-black hole and black ring solutions. The expanding behaviour of the bubbles provides the force necessary to balance the gravitational attraction among the black holes, and hence to reach the equilibrium.Finally, we construct a solution that represents a black hole embedded in a “swirling” universe, which describes a spacetime whirlpool. Moreover, we discuss the possibility of implementing the swirling background in order to enforce the spin-spin configuration, and reach an equilibrium configuration in a double-Kerr spacetime.
BLACK HOLES AND SOLUTION GENERATING TECHNIQUES / A. Vigano' ; supervisore: S. Klemm ; cosupervisor: M. Astorino ; director of the school: M. Paris. Dipartimento di Fisica Aldo Pontremoli, 2022 Oct 25. 35. ciclo, Anno Accademico 2022.
¿SOCIAL CLAUSES IN OUTSOURCING PROCESSES. SOCIAL RIGHTS VERSUS ECONOMIC FREEDOMS IN THE PRISM OF THE MULTILEVEL LEGAL ORDER¿ (¿LE CLAUSOLE SOCIALI NEI FENOMENI DI OUTSOURCING. DIRITTI SOCIALI VERSUS LIBERTÀ ECONOMICHE NEL PRISMA DELL¿ORDINAMENTO MULTILIVELLO¿)
The aim of the thesis is to study the different provisions included in the notion of social clause and offer an in-depth and systematic analysis of some types of social clauses in the Italian and European Union legal order, their characteristics, their regulation, and the legal interests that are aimed at protecting, as well as their classification in the multilevel legal order. This study is preliminary for the assessment on the legitimacy of such clauses in relation to economic freedoms. Indeed, the application of social clauses is one of the fields in which the contrast between fundamental social rights and economic freedoms emerges most clearly and in relation to which some differences occur in the reconciliation of interests made by the Courts at national and EU level. The most relevant distinction for this research relies on the classification developed by Italian scholars, who, on the basis of contents and interests protected by social clauses, usually distinguish between equal treatment or first-generation social clauses and rehiring or second-generation social clauses. Also the nature of the procurement contract, public or private, may condition the judgment on the legitimacy of social clauses with regard to economic freedoms, due to the different regulations applicable in the two fields and the tension between the protection of competition and social objectives, which historically characterizes the action of the public administration in the field of public procurements. The thesis investigates the legislation and the case law concerning the first- and second-generation social clauses, both statutory and contractual, at European and Italian level, with reference to public and private procurements. From this study, it is clear that there are problems of effectiveness and applicability concerning first- and second-generation social clauses, as well as of conflicting interests: social clauses may hinder the competition, negatively condition the entrepreneur’s freedom to conduct a business, affect the entrepreneurs’ freedom of association, and, therefore, risk to contrast with the constitutional and European provisions protecting those freedoms. In this reasoning, the different levels of protection and the importance recognised to social rights and economic freedoms in the Italian and European Union legal systems, the legal framework generated by their interaction, and the type of balancing in the two legal orders must be considered. In the search for a reasonable balance between economic freedoms and social rights, the thesis explores whether the various interests protected by the different types of social clauses can fall within that notion of employment protection which constitutes an overriding reason relating to the public interest justifying a limitation of the economic freedoms and investigates to what extent the prevention of unfair competition and social dumping, the protection of workers’ rights and employment stability can justify a restriction of fundamental economic freedoms. In conclusion, the aim of the study is to investigate if it is possible to achieve a fair balance between opposing interests, social rights, on the one hand, and economic freedoms and free competition, on the other, and comprehend how the interests contrasting with them in relation to the case of social clauses can be balanced.
¿SOCIAL CLAUSES IN OUTSOURCING PROCESSES. SOCIAL RIGHTS VERSUS ECONOMIC FREEDOMS IN THE PRISM OF THE MULTILEVEL LEGAL ORDER¿ (¿LE CLAUSOLE SOCIALI NEI FENOMENI DI OUTSOURCING. DIRITTI SOCIALI VERSUS LIBERTÀ ECONOMICHE NEL PRISMA DELL¿ORDINAMENTO MULTILIVELLO¿) / G. Marchi ; supervisor: M. T. Carinci, L. Ratti ; chairman: F. Scarpelli. Università degli Studi di Milano, 2020 Jan 20. 32. ciclo, Anno Accademico 2019. [10.13130/marchi-giulia_phd2020-01-20].
ROLE OF POLO KINASE CDC5 IN DNA DAMAGE PROCESSING AND REPAIR PATHWAY CHOICE.
Polo-Like Kinases (PLKs) are master regulators of cell cycle in all eukaryotes, and they have also been implicated in the maintenance of genomic stability. Importantly, cell cycle progression is known to affect DSB repair pathway choice and efficiency, but molecular details of these regulations are still mostly unknown. In this thesis, I report the novel characterization of Cdc5, the only PLK in budding yeast, in DSB repair pathways. To this aim, I employed the hypomorphic cdc5-T238A allele in yeast strains which repair a single HO-DSB through specific pathways. The data collected demonstrate that Cdc5 kinase activity is fundamental in promoting Break-Induced Replication (BIR), a detrimental Homologous Recombination subpathway in which a migrating replication bubble elongates a persistent D-loop for tens of kilobases. In particular, cdc5-T238A cells display defective BIR initiation, as they fail to start DNA synthesis from the D-loop. Consistently, cdc5-T238A allele displays reduced Rad51-ssDNA presence at the invading locus, but is proficient in loading Rad51 on ssDNA. The results obtained also exclude the BIR defect to be caused by deregulated resection or impaired DNA damage adaptation in cdc5-T238A mutants. Deletion of Srs2 helicase/translocase completely suppresses cdc5-T238A defect in D-loop extension and double deletion of Srs2 and the Rad51 cofactor Rad55 eliminates the need of a functional CDC5 allele in BIR. Finally, I report the novel identification of Two-Hybrid interactions between Cdc5 and Rad51, Rad55, Rdh54 and Srs2, critical factors in presynaptic filament maturation and D-loop metabolism. In conclusion, this thesis expands the knowledge on PLKs role in DNA repair. Notably, PLKs are commonly deregulated in many tumours and their inhibition is object of study in cancer therapy. The discovery of Cdc5 involvement in BIR proposes a link between PLKs deregulation and BIR-related chromosomal rearrangements observed in cancer cells. Finally, as part of a side project, I exploited Srs2 separation-of-function mutants to investigate Srs2 involvement at different steps of BIR.
ROLE OF POLO KINASE CDC5 IN DNA DAMAGE PROCESSING AND REPAIR PATHWAY CHOICE / M. Di Terlizzi ; supervisore: A. Pellicioli ; coordinatore: S. Ricagno. Dipartimento di Bioscienze, 2025 Feb 20. 36. ciclo, Anno Accademico 2022/2023.
SALT EXPOSURE AND NITROGEN METABOLISM IN TOMATO (SOLANUM LYCOPERSICUM L.): IMPROVEMENT OF THE RESILIENCE TO ABIOTIC STRESS
Salt stress is a critical challenge to plant growth and global food security, necessitating an in-depth understanding of the mechanisms plants employ to adapt to saline environments. Nitrogen (N) metabolism plays a pivotal role in regulating plant responses to salinity, with complex interactions between salt stress and nitrogen availability influencing plant physiology and metabolism.This PhD research explored these interactions through three interconnected studies. The first study examined the physiological and biochemical responses of tomato plants (Solanum lycopersicum L.) to salt stress under varying nitrate (NO₃⁻) supplies. Salinity disrupted the Na⁺/K⁺ balance, reduced NO₃⁻ levels, and modulated N metabolism, as evidenced by changes in amino acids, proline, and GABA levels. The study highlighted the positive effects of higher NO₃⁻ availability, including increased accumulation of osmoprotectants and improved nutritional status, offering valuable parameters for developing salt-resistant cultivars.The second study investigated the differential effects of NaCl and CaCl₂ on tomato plants. While Na⁺ toxicity induced the synthesis of osmoprotectants like proline, trehalose, and GABA, Cl⁻ reduced the availability of key anions and carbon compounds, affecting biomass and Krebs cycle intermediates. These findings underscore the distinct impacts of specific ions on N and C metabolism under salt stress, advocating for tailored agronomic strategies.The third study focused on the potential of a combined NO₃⁻/NH₄⁺ regime to mitigate salt stress. The results revealed that partially replacing NO₃⁻ with NH₄⁺ in nutrient solutions enhanced N assimilation, improved ion balance, and promoted metabolic adjustments in roots and leaves. This dual nitrogen strategy alleviated salinity-induced reductions in growth and development by conserving energy for stress-responsive mechanisms.Overall, this research provides novel insights into the physiological and metabolic responses of tomato plants to salt stress, emphasizing the importance of nitrogen form and availability. These findings contribute to developing agronomic practices and molecular tools to enhance salt tolerance in crops, paving the way for sustainable agriculture in saline environments.
SALT EXPOSURE AND NITROGEN METABOLISM IN TOMATO (SOLANUM LYCOPERSICUM L.): IMPROVEMENT OF THE RESILIENCE TO ABIOTIC STRESS / G. Galli ; tutor: L. Espen, F. Araniti ; coordinatore: S.R. Pilu. - Milano. Università degli Studi di Milano, 2025 Feb 03. 37. ciclo, Anno Accademico 2024.
TARGETING EGFR/ERK/FOS SIGNALING AS A NOVEL APPROACH FOR HPV-POSITIVE HEAD AND NECK CANCER TREATMENT
Squamous cell carcinoma of the head and neck (HNSCC) is the 6th most common cancer worldwide. It accounts for 600.000 new cases and 350.000 deaths every year. HNSCC can be subdivided into two distinct subgroups: HPV-positive and HPV-negative. They present very different features including etiological agent, age, socioeconomic status, prognosis, genetic landscape, and tissue differentiation. Nevertheless, all HNSCC patients are treated with the same therapies, comprising chemo- and radiotherapy, and/or surgery. Specific therapies for HPV-positive HNSCC are needed. HPV oncoproteins E6/E7 are the drivers of oncogenesis in HPV-related cancers and they can be used as targets for therapy. To this end, we studied the effects of the knock down of E6/E7 in HNSCC-positive cell lines and we looked for pharmacological treatments to downregulate E6/E7 levels. We found that targeting the EGFR/ERK/FOS signaling pathway induced a transcriptional reduction of E6/E7 and that the ERK inhibitor Ravoxertinib was effective in killing HPV-positive HNSCC cell lines. Thus, this is a potential novel approach to address HPV+ HNSCC cancer.
TARGETING EGFR/ERK/FOS SIGNALING AS A NOVEL APPROACH FOR HPV-POSITIVE HEAD AND NECK CANCER TREATMENT / A. Medda ; tutor: S. Chiocca ; phd coordinator: G. Viale. Dipartimento di Oncologia ed Emato-Oncologia, 2021 Mar 30. 32. ciclo, Anno Accademico 2020. [10.13130/medda-alessandro_phd2021-03-30].
LA MAFIA 'OLTRE' LA MAFIA: L'ART.416-BIS IN UNA PROSPETTIVA 'DE JURE CONDENDO'
The thesis concerns a proposal for normative modification of Article 416-bis of the Criminal Code,with a focus on the normative technique of normative indices. The structure of the work is composedas follows. The first chapter aims to provide a comprehensive overview of the most recentjurisprudential applications of art. 416-bis to the phenomenon of “new mafias,” highlighting itspeculiarities and the degree of deviation from the traditional approach of mafia-type associationintroduced in 1982. The second chapter starts from a reconnaissance about the profiles ofconstitutional criticality of art. 416-bis, under the twofold profile of determinacy and offensiveness,highlighted by the jurisprudential application mentioned above. It also investigates two phenomenathat are in some ways contiguous (the terrorist phenomenon and the phenomenon of laborexploitation), to highlight how the legislator has intervened normatively in order to go along withtheir profound sociological evolution. In conclusion, the third chapter deals with the “de jurecondendo” perspective, verifying the extendibility of the solutions adopted to the phenomenaassimilated to mafia and proposing the formulation of some normative indices, which appear to theauthor to be the only ones capable of overcoming the criticism in terms of determinacy andoffensiveness. Thereby we ensure an adaptation of the normative instrument to new protection needsconnected to the constant change of the criminal phenomenon.
LA MAFIA 'OLTRE' LA MAFIA: L'ART.416-BIS IN UNA PROSPETTIVA 'DE JURE CONDENDO' / E. Cipani ; tutor: A. P. A. Della Bella ; co-tutor: L. Masera ; coordinatrice: F. Poggi. - Milano. Università degli Studi di Milano, 2024 Jul 12. 36. ciclo, Anno Accademico 2023.
INTRA-TUMORAL INFILTRATION OF GZMKHIGH CD8+ T EFFECTOR MEMORY CELLS IS ASSOCIATED WITH POOR CLINICAL OUTCOME IN NON-METASTATIC COLORECTAL CANCER.
As with most solid tumor malignancies, CRC possess more than one major histological subtype that likely generate disparate immune responses. The heterogeneity in the immune cell response to CRC highlights the need to develop novel immune diagnostics that could predict the risk of disease relapse and devise a secondary strategy to address that risk. The purpose of this study was to prospectively provide a clear description of the patient-specific immune landscape present in CRC to target the dominant immune suppressive factors within a given tumor that may improve response rates while ushering in the age of personalized therapies for cancer patients. We described a functional interplay between neutrophils and CD8+ T cells that impacts on tumor immune escape and relapse. Specifically, we showed that tumor infiltrating neutrophils expressing high levels of CD15 interact with CD8+ T effector memory cells skewing them to produce GZMK, associated with tumor progression in CRC patients. Our fundings identify a unique immune signature, which might inform therapeutic decision making, possibly leading to new immunotherapeutic targets.
INTRA-TUMORAL INFILTRATION OF GZMKHIGH CD8+ T EFFECTOR MEMORY CELLS IS ASSOCIATED WITH POOR CLINICAL OUTCOME IN NON-METASTATIC COLORECTAL CANCER / S. Tiberti ; internal advisor: S. Casola ; external advisor: S. Sdelci ; tutor: T. Manzo ; co-tutor: S. Santaguida ; phd coordinator: S. Minucci. Dipartimento di Oncologia ed Emato-Oncologia, 2023 Apr 13. 34. ciclo, Anno Accademico 2022.
MOLECULAR CHARACTERIZATION OF GENES INVOLVED IN THE PRESENCE OF ANTHOCYANINS IN EGGPLANT (S. MELONGENA L.)
Eggplant (Solanum melongena L.) is a berry-producing vegetable belonging to the large Solanaceae family. Eggplant berries are rich in important secondary metabolites like phenolic compounds (chlorogenic acid in the flesh and anthocyanins in the peel) and flavonols, two classes of antioxidant molecules with suggested health-promoting effects in humans. Anthocyanins pathway is one of most investigated and well characterized pathway among the higher plants. However, the information available for eggplant anthocyanins pathway are rather limited whit the respect to other Solanaceous crop like tomato (S. lycopersicum ) and potato (S. tuberosum). As part of this Ph.D. project, focusing on the highly conserved MYB-bHLH-WD (MBW) transcriptional complex model, which is pivotal in the transcriptional regulation of many anthocyanins related genes, the eggplant MBW were identified and functional characterized. Moreover, a regulatory R3 MYB type repressor (Smel- MYBL1), never reported before, was identified and characterized as well. Fruit peel pigmentation of different varieties of eggplant can vary widely from white to green to purple and violet; in particular, the black/dark purple and dark/light violet peel pigmentations reflect the alternative presence of two different anthocyanins: delfinidine-3-rutinoside (D3R) and delphinidin-3-[p-coumaroylrutinoside]-5- glucoside (nasunin), respectively. The conversion of D3R into nasunin is still an unelucidated part of the anthocyanin pathway in eggplant. The genes responsible of this metabolic step, the SmAAT was identified and functionally characterized through complementation in genotypes carrying the recessive mutant aat allele. Moreover, another genes, Sm5GT1, putatively involved in this step were identified, but more efforts are needed to characterize it. Furthermore, the ripening process is a pivotal aspect which affects the anthocyanins content. The eggplant ripening is commonly divided into three stages: unripe (stage A), commercially ripen (stage B) and physiologically ripen (stage C). Anthocyanin pigments are accumulated to the highest levels at the unripe fruit stage and their levels dramatically decreased as ripening progresses (Mennella et al., 2012), although the total phenolic content remained quite stable (Esteban et al., 1992) leading to the hypothesis that some other still undetermined phenolics are produced during this stage instead of anthocyanins. Focusing on these evidences a list of phenols never reported before in the eggplant peel were identified by HPLC-DAD- UV-VIS analysis. The identification of those new compounds and the recent availability of eggplant genetic resources have allowed the identification of the genes putatively involved in the ripening metabolism of the eggplant berries. The characterization by RT-qPCR three genes were carried out: SmFLS and a couple of SmGT. This genes are now the best candidate for future experiments aimed at irrefutably proving this function.
MOLECULAR CHARACTERIZATION OF GENES INVOLVED IN THE PRESENCE OF ANTHOCYANINS IN EGGPLANT (S. MELONGENA L.) / F.e. Florio ; supervisor: A. Ferrante ; co-supervisor: G. Rotino ; coordinator: D. Bassi. Dipartimento di Scienze Agrarie e Ambientali - Produzione, Territorio, Agroenergia, 2021 Apr 20. 33. ciclo, Anno Accademico 2020.
DISCLOSING THE ROLE OF ROOT EXUDATES IN MODULATING THE INTERACTION BETWEEN PLANT AND XENOBIOTIC DEGRADING BACTERIA
Positive interactions between plants and root-associated bacteria are fundamental to maintain the homeostasis of the holobiont, especially upon environmental stresses. This crosstalk is mediated by root exudates (REs), including plant secondary metabolites, that are released by the plant as a ‘cry-for-help’ to recruit beneficial bacteria. Plant-bacteria interactions are particularly relevant for rhizoremediation, a sustainable strategy for the clean-up of recalcitrant xenobiotics like polychlorinated biphenyls (PCBs), contaminants that induce severe phytotoxic effects, as well as posing serious threats to ecosystems and human health. The aim of this PhD thesis was to disclose the role of root exudates differentially released under PCB stress, in influencing bacterial rhizocompetence traits, necessary to guarantee efficient root colonization and induce PCB degradation.Among root exudates, flavonoids are well described as signaling molecules and were reported as inducers of the PCB degradation pathway. The first objective of this thesis was to test this capability using the PCB degrader Paraburkholderia xenovorans LB400, by assaying pure flavonoids and the complex blend of REs released by Arabidopsis thaliana. LB400-Arabidopsis interaction was monitored in presence and absence of PCB-18 using a fluorescence-tagged LB400 strain, to observe root colonization and plant-growth-promotion potential upon stress. Our findings indicated that flavonoids stimulated LB400 proliferation, induced PCB degradation potential and increased biofilm formation, swimming motility and chemoattraction, features overall implicated in the early root colonization. However, other non-flavonoids molecules seemed to be involved in later stages of colonization and supported the plant growth promotion effect induced by LB400 under phytotoxic growth conditions. The REs alteration caused by PCBs was therefore explored to identify the main players in the contaminant-driven ‘cry-for-help’. A metabolomic analysis was performed on the REs of Arabidopsis grown in control conditions and in presence of a phytotoxic concentration of PCB-18, to observe the differential abundancies of secreted compounds and assess the role of five identified metabolites on the metabolism of PCB-degrading bacteria. PCB-18 dramatically affected plant physiology and altered Arabidopsis exudation pattern. Remarkably, the secretion of scopoletin, a plant secondary metabolite with known antimicrobial characteristics, was inhibited in presence of PCB stress, thus putatively remodeling the rhizosphere ecological niche to provide more suitable conditions for PCB-degrading bacteria. Instead, over-exuded metabolites were utilized as carbon and nitrogen sources and affected different rhizocompetence features. Finally, a bacterial biosensor was developed to clarify the role of root exudation by using a more realistic plant-bacteria interaction experimental set-up. The use of Pseudomonas JAB1 biosensor strain allowed the prompt visualization of root colonization and of the spatial distribution of cells in which PCB degradation was triggered by the REs. Our findings substantiated the conclusions about a putative ‘cry-for-help’ of the plant exposed to PCB-18 and about the centrality of flavonoids in sustaining this beneficial interaction.To conclude, the results of this thesis unveiled the functioning of plant-bacteria associations upon PCB stress and confirmed the relevance of REs in assembling a rhizosphere microbiota that provides the plant with useful services to improve contaminant degradation and support the holobiont health. These findings will be crucial to develop improved rhizoremediation interventions in PCB contaminated soils.
DISCLOSING THE ROLE OF ROOT EXUDATES IN MODULATING THE INTERACTION BETWEEN PLANT AND XENOBIOTIC DEGRADING BACTERIA / E. Ghitti ; tutor: S. Borin ; coordinatore: D. Mora. Dipartimento di Scienze per gli Alimenti, la Nutrizione e l'Ambiente, 2024 Mar 26. 36. ciclo, Anno Accademico 2023.
HYBRID DEEP LEARNING AND RADIOMICS MODELS FOR ASSESSMENT OF CLINICALLY RELEVANT PROSTATE CANCER
Precision medicine holds the potential to revolutionize healthcare by providing every patient with personalized treatments and decisions tailored to his or her individual needs. This might be enabled by the large influx of potentially diagnostic information from new sources such as genetics and modern imaging techniques, provided the relevant information can be extracted. One such framework that has started to demonstrate promise in radiology, especially in the assessment of cancer, is radiomics; the practice of characterizing images by extracting a substantial amount of quantitative mathematical descriptors. This success has largely been enabled by artificial intelligence (AI) and machine learning developments that are capable of handling the big data arrays. Using radiomics, researchers have been able to build prediction models capable of assisting and informing doctors in important decisions such as risk assessment or the choice of treatment. But even though radiomics has shown promise in preliminary studies, there is still a long way to go before radiomics and related AI applications can become routine tools in clinics. The road from patient admission to release is long, and all its intricate steps need to be studied in detail to establish the AI models' benefits and safety. Deep learning is an incredibly powerful AI technique that has revolutionized many areas of science and industry such as recommender systems and protein folding. The technique has demonstrated particular capabilities in image analysis, such as the ability to drive cars autonomously and generate realistic-looking images from scratch. However, the recent advances in deep learning have largely been segregated from the radiomics domain, even though they can synergize with radiomics by performing complementary tasks such as image segmentation and denoising. There is considerable potential for DL and radiomics to cooperatively reinforce each other that so far has been majorly unexplored.This thesis investigates the application of radiomics and deep learning in the context of prostate cancer. It focuses on the clinical perspective of where machine learning implementations are most likely to have a beneficial real-world impact. A key contribution is the deployment aspect: the models are not simply proofs of concept but are conceived and applied in a practical scenario, from patient admission to treatment decision. The specific areas studied include automatic organ segmentation in medical images, automatic quality assurance of segmentations, image processing, and radiomic feature analysis. Finally, a comprehensive study is performed on predicting essential pathological variables with AI, which so far has not been studied previously. Taken together, the methods outlined in this thesis constitute a concrete pathway of how AI can be used to bolster the steps along the patient's clinical trajectory. Successful applications of these methods hold the potential to reduce the workload of clinicians and improve patient outcomes.
HYBRID DEEP LEARNING AND RADIOMICS MODELS FOR ASSESSMENT OF CLINICALLY RELEVANT PROSTATE CANCER / L.j. Isaksson ; internal advisor: S. Gandini ; external advisor: A. Bhalerao ; phd coordinator: S. Minucci. Dipartimento di Oncologia ed Emato-Oncologia, 2022 Dec 16. 34. ciclo, Anno Accademico 2022.
NUTRITIONAL QUALITY AND FOOD SAFETY OF FOOD SOURCES AND FOOD PROCESSING BY-PRODUCTS
In the area of functional foods and dietary supplements, peptides that confer health benefits, also called bioactive peptides, are increasingly recognized as useful tools for improving health and preventing chronic diseases (Udenigwe and Aluko 2012). In fact, food proteins do not only supply nutrients, but also provide numerous health benefits through their impact on specific biochemical pathways. The importance of plant proteins has been underlined by numerous clinical studies performed on grain protein. Most of these activities are due to peptides encrypted in the parent protein sequences, which are delivered by digestion, absorbed by intestinal cells, and transported to the target organs, where they can exert their biological activity (Rutherfurd- Markwick 2012). Actually, bioactive peptides have become a hot research topic, since they provide a large spectrum of health benefits with minimal side effects compared to other chemicals. These health benefits are particularly in the area of hypercholesterolemia and hypertension prevention. Plant-based diet and plant proteins are becoming more and more important to meet nutritional requirements of the growing human population and simultaneously to reduce the negative impacts of food production on the environment. With the aim to boost the environmental sustainability, the valorization of byproducts is certainly one of the main challenges. Some food by-products, such as those deriving from oat, rice, almond, soy okara and coconut processing, are currently among the most interesting by-products of food industries due to their high nutritional potential. Another issue I have addressed in my thesis is the valorization of underutilized species of the genus Lupinus. belonging to the Fabaceae family, whose seeds are characterized by a high protein content. In particular, I have focused the attention on some wild Mexican lupin beans. In fact, a recent review collects data on the nutritional and bioactive compounds of these plants presenting data on proteins, lipids, minerals, dietary fiber, and other bioactive compounds (Ruiz-Lopez, Barrientos-Ramirez et al. 2019), but a characterization of the protein profile is still pending. The information of the protein profile of wild Lupinus species will be beneficial for fundamental and applied studies concerning the storage proteins of the seeds. Based on these considerations, the main aims of my PhD thesis were focused on the discovery, quantification and functional analysis of plant proteins or peptides. These objectives were reached by use of different tools, comprising advanced analytical techniques, such as mass spectrometry (MS) coupled to biochemical and bioinformatics tools, in order to improve the understanding of their mechanism of action at cellular level, their intestinal absorption profile as well as to forecast their potential bioactivities.Briefly, the main aims of the present study were: • To evaluate whether the treatment with some food-grade enzymes, such as amylase, cellulase/xylanase, protease, and their combination, may be useful to achieve this goal. • To valorizing the press cakes deriving from almond and coconut drinks production by using ultrasound- assisted extraction (UAE) to obtain protein ingredients for human use. • To characterize the potential pleotropic activity of two commercially available soybean and pea protein hydrolysates, respectively. • To investigate the composition of the seed proteins of four lupin species: Lupinus aschenbornii, Lupinus campestris, Lupinus hintonii, and Lupinus montanus. The thesis contains three parts:Part I introduces the state of art of proteins and peptides from food and food by-product. Chapter 1 discusses Recovering and obtaining bioactive peptides based on ultrasound-assisted extraction as well as enzymatic methods.Part II presents my scientific contributions in the PhD period.Chapter 2 is the investigat
NUTRITIONAL QUALITY AND FOOD SAFETY OF FOOD SOURCES AND FOOD PROCESSING BY-PRODUCTS / R. Xu ; advisors: C. LAMMI ; A. ARNOLDI. Università degli Studi di Milano, 2023 Mar 27. 35. ciclo, Anno Accademico 2022.
HES CELLS TO STUDY EARLY NEURODEVELOPMENT IN HUNTINGTON'S DISEASE
Huntington Disease (HD) is a genetic neurodegenerative disease caused by a CAG expansion in the gene encoding for the Huntingtin (HTT) protein. The progressive death and atrophy of specific striatal GABAergic projection neurons named Medium Spiny Neurons (MSNs) lead to motor, cognitive and psychiatric dysfunctions in HD patients. Several studies are pointing to the crucial functions of HTT during early development, advocating to a developmental component of HD. Based on these observations, we employed the isogenic series of HD human embryonic stem cells (RUES2) to study the effects of mutant HTT on neuronal differentiation in a fixed genetic background. Firstly, by exposing the control RUES2 cell line to ventral telencephalic differentiation we observed the propensity of the cells to differentiate toward the medial ganglionic eminence (MGE) compared to other human embryonic stem cell (hESC) lines. Moreover, HD RUES2 displayed aberrant MGE cell fate acquisition. Also, these cells recapitulate known HD phenotypes, such as reduced expression of BDNF and NEUROD1, and increased cleavage of N-cadherin. In particular, HD lines exhibit a defect in the transition from pluripotency toward neuroectodermal fate as documented by the persistency of the pluripotent marker OCT4 and by reduced upregulation of the neuroectodermal marker PAX6. Considering the fundamental function of Polycomb group proteins (PcGs) in regulating cell fate identity and differentiation and the crucial role of epigenetics in HD pathogenesis, we started investigating whether these complexes were affected in our cell system. Differences in the number and size of the three-dimensional PcG foci structures emerged between control and HD lines, suggesting a potential link between HTT and these epigenetic complexes. Moreover, level of H3K9me3 histone modification was affected in HD lines during the differentiation. Overall, these preliminary data suggest that the presence of the mutation in the HD gene causes alterations in the regulation of both constitutive and facultative heterochromatin already in pluripotency. This hypothesis will be further tested through experiments of ChIP-seq analysis, in which we expect to identify target genes whose expression may be dysregulated in the early stages of neural development.
HES CELLS TO STUDY EARLY NEURODEVELOPMENT IN HUNTINGTON'S DISEASE / S. Brocchetti ; tutor: E. Cattaneo ; coordinator of the phd school: M. Kater. Dipartimento di Bioscienze, 2022 Jan 13. 34. ciclo, Anno Accademico 2021.
"DEMOCRACY AS A BATTLEFIELD. CONTEMPORARY GRASSROOT MASS MOBILISATIONS IN WESTERN EUROPE, ""POPULAR SOVEREIGNTY"" AND THE REFRAMING OF ""DEMOCRACY"""
How is ‘democracy’ (re)framed in contemporary Western Europe? Which are the consequences of such a (re)framing? Are they desirable? In the conviction that conceptual articulation is dynamic, historical and immanent to political discourses and practices, the starting point of this research is the observation of significant mass grassroot movements in contemporary (2017-9) Western Europe (UK anti-Brexit, French Yellow Vests, Catalan Independentists). The preliminary study is conducted both through direct on field observation and through the collection of public sources (public speeches, press releases, parliamentary debates, political campaigns, newspapers articles, official reports). The research focuses on the debates those movements sparked within their national contexts about the meaning of ‘democracy’. In particular, these debates address the relation between political legitimacy, ‘the Law’ and ‘popular sovereignty’, and the ways in which contemporary ‘democracy’ can be (re)framed within that scope, in the contention between a ‘legalist-constitutionalist’ and a ‘radical-popular’ understanding of it. The first part of the dissertation provides an overview of the theoretical frame within which the research is developed, mainly focusing on performative and immanentistic ontologies and philosphies of language in the ‘New Wittgenstein’ legacy. In the central part of the work, through the application of the Discourse Theory method, the structuration of such debates will be analysed as a political struggle for the hegemony over ‘democracy’ between antagonistic discourses, grounded in different ontologies and values. Each discourse produces conflicting criteria for political legitimation, participation and inclusion/exclusion and different understandings of political spaces, communities and practices, which can escalate in dramatic conflict. One of the most critical differences regards the relation between the acts of foundation and Constitution of a political community, popular sovereignty and the legitimacy of political practices. Those specific empirical cases, as they explicitly address those issues, are, on the one hand, real examples of a contemporary reframing of the concept of ‘democracy’ and, on the other hand, an opportunity for highlighting pivotal political and philosophical questions. The last part will be a tentative elaboration on the consequences of the reintroduction of a radical understanding of ‘popular sovereignty’ for political stability, inclusion, participation, equality and freedom, of their desirability and of the possibilities for those conflicts to be solved.
"DEMOCRACY AS A BATTLEFIELD. CONTEMPORARY GRASSROOT MASS MOBILISATIONS IN WESTERN EUROPE, ""POPULAR SOVEREIGNTY"" AND THE REFRAMING OF ""DEMOCRACY"" / G.a.r. Visintini ; supervisor: A. Besussi ; PhD coordinator: M. Jessoula. Università degli Studi di Milano, 2024 Jan 30. 35. ciclo, Anno Accademico 2021/2022."
TOOLS AND STRATEGIES FOR THE SYNTHESIS OF PHARMACEUTICAL ACTIVE COMPOUNDS
This dissertation presents an overview of three years of research aimed at applying novel tools and strategies for the synthesis of potential pharmaceutical active compounds. In this context, I commenced my research at the University of Milano under the supervision of Professor Daniele Passarella. My work centered on two main projects involving the design, synthesis, and characterization of new pharmaceutical compounds with potential therapeutic applications. These projects focus on convenience and cost-effective synthetic strategies, as detailed in Chapters 1 and 2. I also had the opportunity to spend six months abroad at ICIQ (Tarragona, Spain) in the laboratory of Professor Arjan Kleij. During this time, I worked on the enantioselective synthesis of chiral γ-amino alcohols, which is presented in Chapter 3, as well as the utilization of carbon dioxide as a green starting material to create novel drug-containing macromolecules for future drug-delivery applications, outlined in Chapter 4. In the final part of my Ph.D. program, I moved to Indena SpA (Settala, Italy) to complete my industrial internship, which was a part of my Ph.D. funding. The research conducted during this period focused on the extraction and purification of a component of vaccine adjuvant mixtures, discussed in Chapter 5.Chapter 1 explores the safracin family, a class of alkaloids known for their significant antibacterial and antitumor properties, which were first isolated from bacteria such as Pseudomonas species. This chapter begins with a detailed NMR characterization of Safracin B, an important intermediate in the synthesis of commercially available anticancer drugs. We then report on the synthetic conversion of Safracin B into Cyanosafracin B and Safracin A, two additional components of the safracin family, along with their structural elucidation achieved through NMR spectroscopy. The findings presented will enhance the overall understanding of the chemical profile of safracins and provide a foundation for further chemical and structural investigations into their biological activities and potential applications in drug discovery. Chapter 2 focuses on the design and synthesis of novel tubulin binders in the frame of the European Joint Doctorate TubInTrain program, which examines microtubule dynamics and their relationship to neurodegenerative diseases. The designed scaffolds, created using time- and cost-efficient synthetic methods, feature an amide core with various warheads intended for covalent targeting of tubulin. Biochemical evaluations and X-ray crystallography structure determinations have revealed promising candidates that can covalently bind to the Todalam site. These encouraging results motivate us to continue optimizing these molecules.Chapter 3 discusses the first project I undertook at ICIQ in the laboratory of Professor Arjan Kleij. Specifically, we present the first general enantioselective synthesis of γ-amino alcohols that contain a tertiary carbon center through a Cu-based propargylic amination reaction. Our protocol features user-friendly reaction conditions for the initial oxidation step, and we have established a reasonable scope for both alkynyl oxetanes and functional aromatic amines. Furthermore, the further derivatization of selected γ-amino alcohols demonstrates the synthetic potential of these chiral building blocks, which could diversify the range of chiral intermediates valuable for future drug discovery and development programs. Chapter 4 deals with the development of an effective method for synthesizing drug-modified six-membered cyclic carbonates. These can subsequently be advanced to polycarbonates through organocatalytic ring-opening polymerization (ROP), allowing for a controllable amount of the drug in the resulting polymer-drug conjugates. Preliminary depolymerization studies conducted by Arjan Kleij’s research group indicate that the free acid form of the
TOOLS AND STRATEGIES FOR THE SYNTHESIS OF PHARMACEUTICAL ACTIVE COMPOUNDS / P. Orlando ; tutor: D. Passarella ; co-tutor: P. Allegrini ; coordinatore: D. Passarella. Dipartimento di Chimica, 2025 Jun 05. 37. ciclo, Anno Accademico 2024/2025.
SLEEPING WHILE AWAKE: A NEUROPHYSIOLOGICAL INVESTIGATION ON SLEEP DURING WAKEFULNESS.
Sleep and wakefulness are considered two mutually exclusive states. The alternation between those two states seems to be a defining characteristic of our life, a ubiquitous phenomenon demonstrated in every animal species investigated so far. However, during the last decade, advances in neurophysiology have blurred the boundaries between those states. The mechanisms of sleep have always intrigued neurophysiologists and great advances have been made over the last century in understanding them: we now know that the defining characteristic underlying sleep activity is a specific pattern of neuronal activity, namely the slow oscillation. The slow oscillation, which is characterized by the periodic alternation between periods of activity (ON-periods) and periods of hyperpolarization and neuronal silence (OFF-periods) is the default mode of activity of the sleeping cortex. This alternation is due to the tendency of neurons to fall into a silent period after an initial activation; such tendency is known as “bistability”. There is accumulating evidence that sleep-like bistability, and the ensuing OFF-periods, may occur locally in the awake human brain in some pathological conditions, in sleep transition, as well as after sleep deprivation. Therefore, to the extent that bistability and OFF periods represents the basic neuronal features of sleep, a paradigm shift is in place: from a neurophysiological perspective sleep can intrude into wakefulness. In this thesis, I explore the fluid boundaries between sleep and wakefulness and investigate their possible implications on the problem of personal persistence over time. Moreover, I study the clinical implications of the intrusion of sleep into wakefulness in patients with focal brain injury due to stroke. Specifically, I aim to: 1) show how the sleep-like bistability can be responsible for the loss of function in stroke patients. This may have implications for understanding the pathophysiology of stroke and helping to foster recovery; 2) establish the basis for a model of local sleep that might be present in the everyday life, id est the sensation of sleepiness. Indeed, sleepiness could reflect islands of sleep during wakefulness; 3) advocate the biological criterion of identity, in which the continuity necessary for maintaining ourselves over time could be represented by never resting activity in the brain.
SLEEPING WHILE AWAKE: A NEUROPHYSIOLOGICAL INVESTIGATION ON SLEEP DURING WAKEFULNESS / S. D'ambrosio ; tutors: M. Massimini, S. Sarasso; coordinatore: A. Pinotti. Università degli Studi di Milano, 2020 Jan 31. 32. ciclo, Anno Accademico 2019. [10.13130/d-ambrosio-sasha_phd2020-01-31].
ESSAYS ON GLOBALISATION: EFFECTS AND IMPLICATIONS FOR INDIVIDUALS
This thesis studies the effects of import competition from China and Eastern Europeon the health and fertility decisions of German individuals working in manufacturing.Individuals are matched with separate measures of exposure to competition from Chinaand Eastern Europe, respectively. To isolate exogenous supply shocks from the origin,instrumental variables for competition from each of China and Eastern Europe are constructed.Results in Chapter 1 suggest that higher import competition worsens individual health viajob displacement, wage decline, shortened employment duration, increased reliance onwelfare and less future orientation, with Chinese import competition affecting individualstwice as much. Health declines as individuals increase their visits to the doctor, exerciseless frequently and have a higher probability of developing chronic illness. Also, there issome evidence that individuals do not tend to become disabled but may be slowly pushedinto chronic illness.Findings in Chapter 2 show that import competition negatively affects the individual’sprobability of having children via reduced earnings, lower satisfaction with personalincome and shortened employment duration. The chapter then investigates effects ofimport exposure by gender. Results show that male and female fertility choices differupon rising import competition. Higher import exposure lowers female earnings andjob autonomy, which in turn generates a lower opportunity cost of work, to the pointwhere having children would become a more rewarding alternative for female workers.By contrast, increased import exposure negatively affects male workers’ fertility throughreduced earnings and employment duration.
ESSAYS ON GLOBALISATION: EFFECTS AND IMPLICATIONS FOR INDIVIDUALS / A.a. Piriu ; Ph.D. Advisor: M. Bratti ; Ph.D. Coordinator: A. Missale. Dipartimento di Economia, Management e Metodi Quantitativi, 2020 Apr 15. 32. ciclo, Anno Accademico 2019. [10.13130/piriu-andreea-alexandra_phd2020-04-15].
EVALUATION OF DIFFERENT ABATEMENT SYSTEMS FOR EMISSIONS OF AMMONIA, PM AND ODOUR IN INTENSIVE LIVESTOCK FARMS
Agriculture in the 21st century faces multiple challenges: it has to produce more food to feed a growing population with a smaller rural labour force, adopt and learn new technologies, satisfy consumers’ changing tastes and expectations, and at the same time adopt more efficient and sustainable production methods to cope with climate change, soil erosion and biodiversity loss. Livestock contribution to greenhouse gases (GHG), ammonia (NH3), and odor emissions is relevant. Thus, the identification and adoption of mitigation strategies for intensive livestock farming is necessary, not only to reduce the environmental impact related to livestock production, but also to help solving the issue related to air pollution. Indeed, GHG, NH3 and odor emission other having a negative effect on the environment, represent a risk for human and animals’ health. Air treatment technologies have the double benefit of abating pollutants emissions and improving air quality inside the barns. Alternatively, considering several aspects i) that animal proteins place a heavy burden on our natural resources, ii) the world population and demanding consumers are rapidly increasing, iii) the amount of agricultural land is limited, there is an urgent need to find alternatives to conventional meat products. Terrestrial invertebrates farming represents an environmentally sustainable solution. Earthworms require less land and water, present lower GHG emissions and higher feed conversion efficiency, moreover they are rich in proteins and essential amino acids. The aim of this thesis was to evaluate different mitigation strategies to reduce GHG, NH3 and odor emissions deriving from intensive livestock farming, also taking in consideration their environmental impact. The Life Cycle Assessment (LCA) methodology was applied to quantify the environmental impact of the strategies proposed. In the first step, livestock contribution to air pollutants emissions was quantified. Agricultural NH3 emissions did not even reduce during lockdown periods imposed by Covid-19 pandemic. Thereafter, odor annoyance issue was deeply analysed. Finally, environmental benefits achievable by applying different solutions (i.e. air treatment technologies, manure additive, earthworms rearing) were assessed. The results in this thesis outlined that the adoption of mitigation strategies is fundamental and promising, but each one presents different abatement efficiency on air emissions. Thus, a combination of measures is often the best solution to control emissions, at the same time also an integrated approach combining different methods and techniques could represent the best solution to depict exhaustively the problem and define a good strategy for its management. In this regard, LCA helps to identify the most important contributors to the environmental impact.
EVALUATION OF DIFFERENT ABATEMENT SYSTEMS FOR EMISSIONS OF AMMONIA, PM AND ODOUR IN INTENSIVE LIVESTOCK FARMS / C. Conti ; tutor: J. Bacenetti ; co-tutor: M. Guarino. Dipartimento di Scienze e Politiche Ambientali, 2022 May 19. 34. ciclo, Anno Accademico 2021.
FUNCTIONAL FEED ADDITIVES FOR ANIMAL HEALTH
World population growth is leading to an increase of the of food from animal origin demand. For the future of the planet and food-production system sustainability some important issues required to be faced. Antimicrobial resistance is one of the most important global concerns cause of thousands of deaths per year and livestock can contribute to the antimicrobial resistance spreading. In livestock, the highest use of antibiotic treatments is recorded in young animals, due to their immature immune system, and the presence of several stress sources that predispose young animals to develop multifactorial diseases, particularly from gastrointestinal origin, that causes important economic losses and antibiotic use. Nutrition is no longer considered as science that only satisfies nutritional requirements but is considered fundamental for its functionality through the supplementation of ingredients and additives which contain bioactive molecules that positively impact on health. Functional feed ingredients and additives possess bioactive characteristics, such as antibacterial and antioxidant activities, capable of contrasting infections and improving animal health or a particular organic function. Preserving intestinal health decreases the incidence of pathologies, optimises digestive processes, and promotes animal performance thus resulting in higher profitability for the farmers. The aim of the following project was to evaluate innovative functional feed additives for animal nutrition, in order to increase animal health, performance and reduce the use of antibiotics in farm animals. The attention was focused on the evaluation of algae, tannins extract, leonardite, tributyrin and probiotics as functional feed additives alternative to antibiotics. For reaching this goal different in vitro, in vivo and ex vivo approaches have been used. In vitro methods allowed to characterize functional feed additives for their antimicrobial and antioxidants effects and cell culture was useful to predict their interaction with the intestinal epithelium. In vivo zootechnical trials were conducted on young animals to assess dietary supplementation of functional additives on growth performance, prevention of gastroenteric disorders, metabolic status and microbiota modulation. Obtained results highlighted that functional additives were characterized by bioactive properties such as antioxidant and antimicrobial effects. Their supplementation showed a plethora of positive impacts including higher zootechnical performance, decreased diarrhoea occurrence, improved health status and microbiota modulation. A combined effect was also assessed in order to evaluate a possible synergistic of different additives. The combination resulted in a positive effect for diarrhoea prevention with a slight modulation for the gut microbial environment. Even if feed additives displayed different bioactivities in vitro, they did not always reveal a boosting effect on growth performance in vivo resulting less evident for farmer perception. However, the reduction of gastrointestinal disorders incidence could be considered the most interesting characteristic of functional feed additives which have shown to possibly reduce pathologies occurrence thus improving farm profitability.Lastly, due to the needing to decrease animal for experimental purposes an innovative ex vivo intestinal perfusion model was developed in order to study nutrient bioaccessibility for obtaining reliable science-based data in line with the 3R principles.In conclusion, tested compounds can not substitute the therapeutic treatment of antibiotics. However, the utilization dietary approaches supplementing functional feed additives can significantly contribute for decreasing pathology occurrence, boosting animal health and performance thus the antibiotics use. In line with the One Health principles, antibiotics reduction is a key point for the future sustainable development on the planet. The use of functional
FUNCTIONAL FEED ADDITIVES FOR ANIMAL HEALTH / M. Dell'anno ; tutor: L. Rossi ; coordinatore: F. Ceciliani. Dipartimento di Medicina Veterinaria e Scienze Animali, 2023 Feb 21. 35. ciclo, Anno Accademico 2022.
VISIBLE LIGHT SENSITISED WO3−BASED PHOTOANODES FOR SOLAR ASSISTED WATER OXIDATION
The need for stable, storable, accessible and renewable forms of energy has been insistently growing in the last decades, along with the concern about the ongoing climate changes connected to the extensive use of fossil fuels. The efficient exploitation of solar energy, that strikes the Earth’s surface with a great amount of energy everyday, is a promising, though challenging, approach to the problem, with hydrogen produced from solar energy using photoelectrochemical water splitting representing an encouraging alternative.This PhD thesis aims at contributing to deepening the knowledge of the working principles and intrinsic characteristics of photoanode materials, responsible for driving the water oxidation reaction, the kinetic bottleneck of overall water splitting. Focus of the work is the study of photoactive metal oxide-based materials, such as tungsten trioxide (WO3), its ternary oxide derivative copper tungstate (CuWO4) and bismuth vanadate (BiVO4), which are among the most promising materials to be employed for these scopes.Most of the experimental work here presented has been conducted at the UniMi Photocatalysis Group of the Università degli Studi di Milano, Italy, under the supervision of prof. Elena Selli; a fruitful collaboration with the group of prof. Sixto Giménez at the INAM Institute of the Universitat Jaume I of Castellón de la Plana, Spain, has led to the results outlined in Chapter 7 and partially in Chapter 5.The present PhD thesis is organized as follows.Chapter 1 presents an introduction to the grounds and aims at the origin of the work. An outline on the global energy demand and consumptions is drawn, evidencing that fossil fuels still represent the most widely used energy source. A brief description of the new, emerging renewable energy sources is presented, with a more detailed view on the hydrogen fuel, in particular on that produced from solar assisted water splitting.Chapter 2 aims at giving the theoretical principles at the basis of photoelectrochemical water splitting. The main category of photocatalysts, i.e., metal oxide semiconductors, is presented along with their physico-chemical properties. The peculiar characteristics that make them suitable photoactive systems for driving the water oxidation reaction are highlighted. Finally, the working mechanism and the requirements of combined tandem PEC cells are illustrated.Chapter 3 is dedicated to both the experimental and theoretical methods, as well as the synthetic and characterization techniques, that have been used throughout my PhD for producing and interpreting the data presented in this thesis. A careful description of the employed experimental set ups, located in the laboratories of both prof. Selli and prof. Giménez, is also reported.Chapter 4 starts with the description of the experimental work carried out during my studies, which was firstly focused on copper tungstate, a ternary oxide deriving from the more investigated tungsten oxide, which is characterized by a relatively smaller band gap enabling its sensitisation towards the visible portion of the solar spectrum. For the first time, the effects on photoactivity induced by doping this material with nickel ions have been systematically investigated and demonstrated by means of multiple physical and photoelectrochemical methods.Chapter 5 deals with the study of the well-known WO3−BiVO4 heterojunction, a system combining two of the most promising photoanode materials. My work was focused on disclosing the effects on the overall performance of the composite material induced by selectively tuning the morphology of the tungsten oxide underlayer. WO3 and WO3−BiVO4 photoanodes characterized by either a planar or a nanostructured morphology have been synthetized and multiple experimental techniques have been applied to ascertain the intrinsic characteristics as well as the photoelectrochemical performances of the so obtaine
VISIBLE LIGHT SENSITISED WO3−BASED PHOTOANODES FOR SOLAR ASSISTED WATER OXIDATION / C. Nomellini ; tutor: E. Selli ; co-tutor: M. V. Dozzi ; PhD course coordinator : D. Passarella. Università degli Studi di Milano, 2023 May 08. 35. ciclo, Anno Accademico 2022.
MULTIFACETED DRIVERS OF PERFORMANCE VARIATION IN ANIMAL POPULATIONS: INVASIVE PREDATOR, RESOURCE AVAILABILITY AND HYBRIDIZATION
The biodiversity on Earth is dramatically affected by global changes. To understand biodiversityloss and determine priorities for conservation, it is important to assess whether and how individualscan adapt to a continuous and drastic environmental change. Phenotypic variation is a key processthat allow populations to withstand environmental changes by increasing survival and fitness, andcan be related to both adaptations and phenotypic plasticity. For this reason, studies on phenotypicvariation, and its associated fitness, analysing response to different stressors are needed. This thesisinvestigated phenotypic variability, through the study of performance variation, in response to threeof the most pressing issue of global change: alien species, resource availability and hybridization.The first study (Chapter 2) focused on phenotypic variation of a native species in response toan alien predator. Italian agile frog embryos developed faster when exposed to the predator inlaboratory condition and when collected in alien crayfish-invaded sites. This ontogenetic shift canbe interpreted as a local adaptation to the alien predator and suggests that frogs are able to recognisethe predatory risk. If these responses are effective, escape strategies against the invasive predatormay improve the survival of frog populations.The second study (Chapter 3) analysed how sexually selected traits vary across differentdegree of resource availability. We assessed the existence of condition- and context-dependentsexual dimorphism (SD) within and among populations in the Italian wall lizard. Body conditionand ecosystem productivity were the main drivers of body size SD variation, and body conditionwas also the main driver for head shape SD. The expression of SD in the Italian wall lizard isstrictly related to the changes in resource availability.The third study (Chapter 4) evaluated how hybridization affects hybrid performance and themain drivers of performance differences between hybrid and their parents, using the meta-analyticapproach to obtain generalizable conclusions. The averaged hybrid performance was similar to the 2fitness of parental lineages, however both the level of genetic divergence between parental lineagesand the approach used to identify hybrids affected hybrid performance. Performance was lower forhybrids between distantly related lineages and the use of imprecise approaches for hybrididentification (e.g. morphology-based) can bias assessments of performance.Altogether, the analyses performed show how alien species, resource availability andhybridization can drastically modify the phenotypic variability of animal populations. The presenceof an alien predator induced plastic and adaptive phenotypic changes in a threatened frog that couldincrease the survival of the populations. Resource availability determined the degree of expressionof sexually selected traits among and within lizard populations. Finally, hybridization can decreasehybrid performance when epistatic interactions occurred in individuals. Here, I demonstrated thatanimal responses to different stressors are particularly varied and articulate. Future challengesshould focus on the combined effects of several stressors on animal phenotype. The conclusions ofthis thesis represent a step toward a better knowledge of the impact of global changes on phenotypictraits variation related to performance in animals.
MULTIFACETED DRIVERS OF PERFORMANCE VARIATION IN ANIMAL POPULATIONS: INVASIVE PREDATOR, RESOURCE AVAILABILITY AND HYBRIDIZATION / M. Muraro ; tutor: G. F. Ficetola ; tutor: S. Sherpa. Dipartimento di Scienze e Politiche Ambientali, 2023 May 02. 35. ciclo, Anno Accademico 2022.
TERNARY OXIDE SEMICONDUCTOR PHOTOANODES FOR SOLAR ENERGY CONVERSION
Solar energy conversion and storage into hydrogen is a valuable approach to capture the energy that is freely available from sunlight and to turn it into a clean fuel. Photoelectrochemical (PEC) water splitting through the dual-absorber tandem cell technology has emerged as a promising strategy to this aim. The work conducted in the frame of this PhD thesis aimed at playing a part in the development and optimization of efficient oxide-based semiconductor photoanodes for water oxidation, which is the kinetic bottleneck of the overall PEC water splitting process. Photoanodes based on films of absorbing materials were successfully synthesized with a high optical transparency as important requirement for maximizing the solar energy conversion efficiency of the final tandem cell device. Subsequently, their intrinsic properties as single photoabsorber photoanodes were largely improved, on the basis of the results obtained through comprehensive PEC studies in parallel with thorough structural, morphological, and spectroscopic investigations.The attention was focused on three different classes of promising ternary metal oxides, able to absorb a large portion of the solar spectrum, namely i) BiVO4 (bandgap Eg = 2.4 eV), known for its excellent solar light to hydrogen conversion efficiency, ii) the copper tungstate-based materials CuWO4 (Eg = 2.3 eV) and CuW(1-x)Mo(x)O4 (Eg = 2.0 eV), ideal to be employed as visible-light active alternative to WO3, and iii) ZnFe2O4 (Eg = 2.0 eV) belonging to the spinel ferrites class, possessing excellent photothermal and chemical stability. Specifically, BiVO4 was studied either as a visible light sensitizer towards TiO2 or as a single photoanode material to focus on the identification and improvement of its intrinsically poor electron transport and interfacial transfer properties. In the first case, the TiO2/BiVO4 heterojunction system was proved to be effective in producing highly reductive electrons, suitable for overall water splitting, through TiO2 sensitization towards visible light. This, together with the counterintuitive mechanism at the basis of the observed impressive functionality, was effectively disclosed through combined PEC and photocatalytic reduction test studies. The multifaceted role of Mo6+ doping onto both the bulk and surface properties of BiVO4 films was also revealed through an in-depth PEC and impedance spectroscopy study. By improving either the bulk conductivity or the interfacial charge transfer of optimized Mo6+ doped BiVO4 photoanodes a conspicuous enhancement was attained of their photoactivity towards water oxidation with respect to the pure material. The presence of intra-gap states in CuWO4, acting as electron traps and thus being responsible for a severe internal charge recombination, was verified by means of the first ultrafast transient absorption study performed with this material, in combination with both an electrochemical and a photochromic characterization. This issue, which strongly limits the PEC performance of CuWO4 photoanodes, was addressed by adopting a 50% Mo for W substitution resulting in CuW0.5Mo0.5O4 photoanodes, exhibiting not only a greatly extended visible light-induced photoactivity compared to the pure material, as a result of enhanced absorption, but also a considerably improved charge separation. All these factors contributed to the much better PEC performance attained with respect to CuWO4 electrodes. This study was finalized by the identification of a suitable hole scavenger species for copper tungstate-based materials, able to ensure enhanced photocurrent generation compared to pure water oxidation while minimizing dark currents.Finally, in the frame of my seven months stage in Prof. Sivula’s group at the EPFL in Lausanne, a thorough study was performed on the impact that several parameters, such as the annealing temperature, the film thickness and the creation of oxygen vacancies
TERNARY OXIDE SEMICONDUCTOR PHOTOANODES FOR SOLAR ENERGY CONVERSION / A. Polo ; tutor: M. V. Dozzi ; co-tutor: E. Selli ; coordinatore: E. Licandro. Dipartimento di Chimica, 2021 Mar 24. 33. ciclo, Anno Accademico 2020. [10.13130/polo-annalisa_phd2021-03-24].
SUSTAINABLE PRODUCTION AND PHYSICO-CHEMICAL CHARACTERIZATION OF BIOSURFACTANTS FROM RENEWABLE RESOURCES
Cheese whey permeate (CWP) represents the main waste stream of the dairy industry. To produce 10 kg of cheese, around 100 kg of milk are used and roughly 90 kg of cheese whey (CW) are generated as a by-product. CW is still considered a valuable source of milk proteins, which are recovered by ultrafiltration, thus resulting in around 89 kg of CWP, but it contains a high concentration of lactose (up to 45-50 g/L). In the frame of applying a bio-based circular economy approach, this study aims at exploring the use of lactose and its hydrolysis products, glucose and galactose, as cheap and abundant substrates to produce value-added products, like sugar fatty acid esters (SFAEs). SFAEs are non-ionic surfactants that are characterized by excellent surface and interfacial tension reduction capability, low toxicity, and biodegradability. Moreover, the components of SFAEs can derive from natural resources, thus allowing the final product to be labeled as natural and bio-based as well. These features make them extremely promising for industrial applications as emulsifiers in the cosmetic and food sectors. Some SFAEs are currently produced at industrial level and they are available on the market (i.e. sorbitan and ethoxylated sorbitan esters, Span® and Tween®, as well as sucrose esters and alkyl polyglycosides). However, their current synthesis still rely on acid homogeneous catalysis and harsh reaction conditions, which result, in most cases, in complex mixtures of products with different degrees of esterification and different acylation positions, as well as mixtures of undesired by-products, deriving from side reactions, such as sugar dehydration/caramelization. In this thesis, alternative and more sustainable approaches have been described. In particular, enzymatic esterification, acetalization or Fischer glycosylation followed by solvent-free esterification, were investigated as sustainable alternatives to the currently established chemical schemes for the preparation of a small library of performant SFAEs, by exploring the use of challenging substrates, i.e., lactose and CWP. Moreover, the physico-chemical properties, such as interfacial tension features, W/O emulsification capability and the relative stability over time, of the prepared biosurfactants were evaluated to assess their potential application as bio-emulsifiers in the food, pharmaceutical and cosmetic industries.
SUSTAINABLE PRODUCTION AND PHYSICO-CHEMICAL CHARACTERIZATION OF BIOSURFACTANTS FROM RENEWABLE RESOURCES / S. Sangiorgio ; tutor: G. Speranza ; co-tutors: G. Cappelletti, D. Ubiali. Dipartimento di Chimica, 2023 Apr 17. 35. ciclo, Anno Accademico 2022.
CONDIZIONI GENERALI DI CONTRATTO E TUTELA DELLA CONCORRENZA
The present thesis seeks to inquire the possibility to operate a control on the content of standard contracts through the application of competition law: namely, it aims to identify in which cases an enterprise violates competition law by adopting standard terms and employing them in its contractual relationships; furthermore, it wishes to analyze which remedies are available to the enterprise’s contractual counterparties. The enquiry is particularly relevant in the current socio-economical context, in which new market powers are rising and seem able to defy traditional antitrust law tenets. The reference is to the so-called “Big Tech companies”, i.e. firms (such as Facebook, Google or Amazon) which, by offering apparently free services (which are actually paid for by the exploitation of users’ personal data), have hitherto managed to escape antitrust scrutiny. However, some recent investigations have reversed the trend and have found these firms guilty of infringing antitrust law by adopting terms and conditions particularly prejudicial to the user (especially with regards to protection of its privacy). From another point of view, the “antitrust control” on standard contracts may allow to identify the remedy available to the prejudiced party within contractual relationships where, despite the fact that both parties are technically “professional”, a significant imbalance of bargaining power exists, so that the weak party is forced to enter into a detrimental contract. In order to answer these questions, the thesis analyses the debate between scholars regarding the feasibility of a control of substance of standard contracts, whereas the Italian Civil Code only provides for a formal control; moreover, it examines the different doctrinal opinions on the matter of the relation between antitrust prohibitions and the contracts that the dominant enterprise stipulates with professionals and consumers alike.
CONDIZIONI GENERALI DI CONTRATTO E TUTELA DELLA CONCORRENZA / M.g. Moresco ; tutor: F. Delfini ; coordinator: M. T. Carinci. DIPARTIMENTO DI DIRITTO PRIVATO E STORIA DEL DIRITTO, 2020 Feb 07. 32. ciclo, Anno Accademico 2019. [10.13130/moresco-matteo-giovanni_phd2020-02-07].
ANALYSING TEACHERS' RELATIONSHIPS TO INVESTIGATE TERTIARY EFFECT AND WORK-RELATED OUTCOMES
The aim of this dissertation is to investigate the influence of teachers' social networks on various aspects of their professional and personal attitudes. In particular, it focuses on teachers' stereotypes and biases, which may represent antecedents of the so-called tertiary effects of social origin on educational inequalities—namely, the often unconscious ways in which schools and their agents contribute to the reproduction of these inequalities. The use of Social Network Analysis (SNA) offers an innovative contribution to this field by exploring how stereotypes and biases are shaped and reinforced through interpersonal relationships, depending on both the characteristics of the individuals within the network and the structural features of the network itself.The dissertation begins with a theoretical overview of tertiary effects and teacher biases, followed by two empirical chapters based on original data collections that address the role of teachers’ social networks. The first study draws on the tradition, relatively underexplored in Italy, of network research among teachers, and investigates how collegial interactions influence job satisfaction and self- efficacy. The second study, which bridges the SNA approach and the literature on tertiary effects, examines how teachers’ networks impact the development and persistence of stereotypes related to educational inequality.The main contribution of this work lies in proposing a relational perspective for analysing teacher biases, offering a novel implementation of Social Network Analysis within this context. The findings highlight the importance of considering teachers’ social networks in the analysis of both their professional outcomes and their role in perpetuating educational inequalities.
ANALYSING TEACHERS' RELATIONSHIPS TO INVESTIGATE TERTIARY EFFECT AND WORK-RELATED OUTCOMES / G. Zanga ; supervisor: G. Ballarino ; co-supervisor: R. Vacca ; coordinatore: M. Guerci. Dipartimento di Economia, Management e Metodi Quantitativi, 2025 Sep 25. 37. ciclo
DEVELOPMENT OF INNOVATIVE INSECT MEALS AS FUNCTIONAL FEED INGREDIENTS FOR MONOGASTRIC ANIMALS
The global population is facing exponential growth, projected to exceed 9 billion by 2050, raising significant challenges for food security and sustainability. This rapid increase drives higher demand for food, particularly animal-based products, placing pressure on the agricultural and livestock sectors Addressing the resulting protein gap requires innovative, sustainable strategies that minimize the environmental impact of conventional feed sources like soybean and fishmeal. Sustainable animal nutrition has thus become a priority, focusing on optimizing resource efficiency, enhancing livestock health and productivity, and reducing antibiotic dependence. Key measures include incorporating alternative feed ingredients, by-products, and recycling nutrients from former feedstuffs. The One Health approach supports these efforts by integrating food safety, zoonotic disease control, and antimicrobial resistance management, recognizing the need for multidisciplinary strategies to support human, animal, and environmental health. In line with these challenges, the United Nations 2030 Agenda for Sustainable Development and the European Green Deal emphasize the importance of sustainable agricultural practices and the reduction of environmental impacts, while ensuring food security.In this context, insects offer a promising, sustainable protein source that can help reduce the reliance on arable land for animal feed crops production. By valorising food waste and agricultural by-products as insect substrates, this approach supports a circular economy, minimizing environmental impact and optimizing resource efficiency. Insect-derived meals offer a high-quality protein source combined with bioactive components like chitin and antimicrobial peptides. These components exhibit prebiotic effects, supporting beneficial bacterial populations and probably supporting intestinal integrity.Among various insect species, Tenebrio molitor (commonly known as mealworm) is notable for its valuable nutritional profile, primarily due to its high biological value proteins and functional components.This doctoral research aimed to advance the understanding of insect-based nutrition by exploring innovative rearing strategies for Tenebrio molitor larvae, with the goal of optimizing larval growth performance, enhancing the nutritional and functional profiles of insect-derived meals, and evaluating their application as a sustainable ingredient in pig nutrition. The first part of research focused on the evaluation of innovative hydration strategies (water with 12.7% of lactoferrin) and circular growth substrates from agro-industrial by-products (chestnut shell, carob, brewer’s spent grain), to optimize rearing methods and larvae performance. Obtained T. molitor larvae meals were analyzed for their nutritional composition (AOAC, 2023), main functional properties (antioxidant activity and inhibitory activity against E. coli strains) and metabolomic profile through nuclear magnetic resonance (NMR) spectroscopy. Obtained results showed that the substrate can influence the growth of T. molitor, as well as the morphological and chemical-functional properties of the resulting insect meals. Particularly, we observed that the inclusion of chestnut shell led to higher growth rate compared the control after 14 days, with values of 7% and 4%, respectively (p=0.0253). Similarly, carob and brewer’s spent grain included at 25% resulted in significantly enhanced growth rates of 12%, compared to 4% in the control (p=0.0395). Moreover, innovative rearing methods impacted the final color of insect meals. Specifically, 12.5% inclusion of chestnut shell showed a color variation (ΔE=5.59) compared to the control, as well as larvae hydrated with lactoferrin solution (ΔE =1.66). Regarding insect nutritional profile, differences in protein content were found, with increased levels registered in larvae reared on chestnut shell (12.5%
DEVELOPMENT OF INNOVATIVE INSECT MEALS AS FUNCTIONAL FEED INGREDIENTS FOR MONOGASTRIC ANIMALS / I. Ferri ; tutor: L. Rossi ; coordinatore: F. Ceciliani. Dipartimento di Medicina Veterinaria e Scienze Animali, 2025. 37. ciclo, Anno Accademico 2023/2024.
NEW INSIGHTS INTO PATHOGENESIS AND PROGNOSIS OF CANINE SARCOMAS
The majority of canine soft tissue and visceral sarcoma prognostic studies have been influenced by a “one size fits all” approach which lead researchers to merge different subtypes of sarcomas in the same study population. The consequent heterogeneity of cases hampered the interpretation of the results per each tumor category and impeded the comparison of the results among different studies and their application in veterinary clinical oncology. The majority of these studies have also been characterized by non-standardized and non-rigorous methods. Given these premises, the aims of this thesis were to identify valuable prognostic factors specific for different canine sarcoma subtypes, following rigorous and repeatable methods, leading to a better definition of patient outcome, as well as provide novel insights into canine sarcoma pathogenesis, providing a basis for improved clinical management and possibly targeted therapy.The introduction provides an overview of classification and prognostic factors in cutaneous and subcutaneous soft tissue sarcomas (STSs), as well as visceral sarcomas, with a special focus on canine perivascular wall tumors (cPWTs) and splenic stromal sarcomas (SSSs).The introduction is followed by the description of four distinct investigations. The first study analysed the impact of clinical and histopathological variables on local recurrence (LR) and overall survival time (OST) in a large series of retrospectively collected surgically excised cPWTs finding that histological grade, tumor necrosis, mitotic count, status of surgical margins, tumor ulceration, and location on distal extremities had an impact on LR and that ulceration, grade III, necrosis >50% and higher mitotic count were correlated with shorter OST. The study results suggested that all these prognostic factors need to be considered to predict cPWTs prognosis more accurately. The second study aimed at evaluating the immunohistochemical expression of survivin and β-catenin in a series of retrospectively collected surgically excised cPWT samples to investigate their possible involvement in the tumorigenesis and progression of these tumors. Additionally, the study attempted to recognize the prognostic impact of survivin, β-catenin, and Ki-67, together with histopathological and clinical variables, on LR and OST. cPWTs variably expressed survivin and β-catenin at both nuclear and cytoplasmic level, and increased nuclear survivin was associated to a reduced OST. Inclusion of survivin immunohistochemical expression analysis when diagnosing cPWTs may allow to better define patient prognosis and to eventually plan adjuvant treatments in selected dogs. Additionally, survivin and β-catenin may be potential therapeutic targets for more aggressive phenotypes of these tumors. Finally, the study confirmed the prognostic role of mitotic count and tumor size in cPWTs.The third study focused on canine STSs surgical margin assessment investigating the impact of clean but close margins (CbCMs) on LR. The study reported that the cumulative incidence of LR at three years differed significantly between infiltrated margins, CbCMs, and tumor-free margins. Canine STSs excised with infiltrated margins had the greatest risk of LR, and the rate of LR with CbCMs was greater than LR rate of tumor-free margins. Both when CbCMs were grouped with infiltrated margins and when CbCMs were grouped with tumor-free margins, a significant difference between groups persisted. Thus, the conclusion was that CbCMs should be considered as a separate prognostic category for canine STSs.The fourth study assessed the impact of pathological and clinical variables on the occurrence of metastasis in 32 cases of canine SSSs, which were classified into different subtypes on the basis of histomorphological and immunohistochemical features. The research has demonstrated a potential prognostic role of the grading system usually applied
NEW INSIGHTS INTO PATHOGENESIS AND PROGNOSIS OF CANINE SARCOMAS / F. Godizzi ; tutor: P. Roccabianca; coordinatore: F. Ceciliani. Dipartimento di Medicina Veterinaria e Scienze Animali, 2023 Jun 09. 35. ciclo, Anno Accademico 2022.
IDENTITÀ RELIGIOSA E DIRITTI DI GENERE. L'IMPATTO DEGLI STATUTI PERSONALI SU BASE RELIGIOSA SULLE DONNE CRISTIANE IN CISGIORDANIA
This doctoral thesis investigates the elements of discrimination that exist within the normative sources of Christian communities in the West Bank. The research analyses the impact of religiously based personal statutes on women in the local communities - in particular the Catholic, Orthodox and Protestant communities. The analysis of the complex legal context of the West Bank aims to highlight both the criticalities present in the positive law itself and the opportunities for change promoted 'from within' the Palestinian Christian communities, in particular the ambitious reform project that has affected the Palestinian Lutheran community in 2014.
IDENTITÀ RELIGIOSA E DIRITTI DI GENERE. L'IMPATTO DEGLI STATUTI PERSONALI SU BASE RELIGIOSA SULLE DONNE CRISTIANE IN CISGIORDANIA / L.a. Bulgar ; tutor: D. Milani, F. Poggi. Dipartimento di Scienze Giuridiche Cesare Beccaria, 2023. 36. ciclo, Anno Accademico 2023.
NETWORKS OF GROUP EQUIVARIANT NON-EXPANSIVE OPERATORS FOR ARTIFICIAL INTELLIGENCE. MODELS, APPLICATIONS AND INTERPRETABILITY.
Artificial Intelligence (AI) and Machine Learning (ML) are increasingly integrated into our daily lives, yet we often encounter opaque, black-box algorithms driving these systems, which can be deceptive or counterfeit. The pursuit of eXplainable Artificial Intelligence (XAI) aims to develop methods that clarify the decision-making processes of black-box AI systems, making them more understandable and trustworthy for end users, in line with regulatory and policy demands.Another significant challenge facing XAI is creating algorithms that are intrinsically explainable by design. In this context, the mathematical theory of Group Equivariant Non-Expansive Operators (GENEOs) has all the credentials to be at the base of a new generation of ML models with desirable geometric properties. This thesis aims to support this claim by investigating GENEO networks from both an application and theoretical perspective, demonstrating their effectiveness as building blocks for competitive networks in terms of performance and explainability. In essence, a GENEO is an operator between two functional spaces that satisfies two important properties: firstly, there is equivariance, which prescribes that the operator must commute with all the elements of a certain group of transformations of the functional data domain, and secondly, non-expansivity, which requires that the distance between the outputs of a GENEO must be always smaller than the original distance between the inputs, when measured with the appropriate distance. From the point of view of applications, I will present in particular GENEOnet, which is the principal implementation of a GENEO network and that was developed during an industrial collaboration with experts in medicinal chemistry on the problem of protein pocket detection. More generally, I will present and analyze several GENEO networks designed to tackle various types of data and problems. These networks outperform other state-of-the-art domain-specific models while also featuring enhanced explainability and robustness properties. The primary experimental finding is that such networks can deliver strong results relying on a minimal number of learnable parameters, making them easier to interpret and study, while moving towards intrinsically explainable models. Furthermore, the reduced number of parameters helps to maintain the model structure simple and decreases computational costs, compared to more complex models like deep neural networks.From a theoretical perspective, I will introduce new techniques for generating linear GENEOs within the context of graph theory, yielding GENEOs that can commute with the group of graph isomorphisms. These methods are based on the concept of permutant and generalize previously developed results, particularly by using input-dependent permutants to embed specific subgraphs into the graph of interest. Using aggregation results for GENEOs, these subgraph search operators enable the definition of a novel isomorphism test for graphs. I compared this test with other conventional approaches in terms of accuracy and computational cost, and it was found to be superior when considering both criteria simultaneously.The findings in this thesis establish the basis and initial experimental evidence for GENEOs as tools for creating networks that belong to the class of models explainable by design. The ultimate objective of this research is to further substantiate this claim by constructing a library of GENEOs easily used for various use cases.
NETWORKS OF GROUP EQUIVARIANT NON-EXPANSIVE OPERATORS FOR ARTIFICIAL INTELLIGENCE. MODELS, APPLICATIONS AND INTERPRETABILITY / G. Bocchi ; tutor: A. Micheletti, P. Frosini, C. Talarico ; coordinatore: G. Ciraolo. - Milano, Dipartimento di Matematica. Dipartimento di Scienze e Politiche Ambientali, 2025 Feb 18. 37. ciclo, Anno Accademico 2023/2024.
CONTESTED TERRITORY: VIOLENCE, ORGANISED CRIME AND TERRITORIAL PRACTICES IN CONTEMPORARY HONDURAS
The relationship between organised crime and territory is a complex one, and one which affects extensively security dynamics. Especially in countries plagued by violence and insecurity, like Honduras, territory is strategically important for criminal actors, that usually seek to obtain territorial control in certain areas and often compete over it with their rivals and authorities. Despite this, Honduras remains a highly understudied country in academic and policy discourses on the issue, especially due to the objectively dangerous nature of the criminal groups present in the country and the elevated levels of corruption and impunity that permeate it. As a result, the ways in which criminal groups shape the country’s territory, affecting local communities and security dynamics, remains an underexplored aspect in the literature. Especially, little is known about the way in which criminal groups’ presence, activities, and interactions shape space and territory across different scales in Honduras.By drawing upon thirty semi-structured interviews with Honduran residents, ex-residents, and experts on criminal groups, activities, and dynamics in the country, and the analysis of official judicial documents, secondary literature, available public reports, and media outlets, this qualitative case study explores the ways in which criminal, state, and non-state actors shape the territory in Honduras in the context of a liberal market democracy. By focusing on different scales, respectively the local, regional, and national, and the interplay among these, this study aims at painting a picture of the bordering and territorial practices that different actors – especially criminal groups – have been carrying out in the country approximately in the last three decades. Adopting an interdisciplinary approach, this thesis consolidates and integrates the literature on organised crime, violence, the state-crime nexus, territory and territorial practices, and borders and bordering practices with original empirical research.By focusing on gangs in urban areas of the country, drug trafficking organisations in rural ones, and state-crime relationships across the national territory, I argue that these actors have been able to actively produce and shape the country’s territory along micro, meso, and national lines, often establishing authority on it, and have contributed to shaping the Honduran territory as a deeply contested space. In doing so, I argue, these groups mirror – and, at the same time, are embedded into – the dynamics of capital accumulation and resource commodification entrenched in the country’s history of capitalist and neo-colonial exploitation and occupation throughout the centuries.
CONTESTED TERRITORY: VIOLENCE, ORGANISED CRIME AND TERRITORIAL PRACTICES IN CONTEMPORARY HONDURAS / E. Ziosi ; tutor: C. Castellano, M. Rosti; coordinatore: F. Basile. Dipartimento di Scienze Giuridiche Cesare Beccaria, 2023 Jun 26. 35. ciclo, Anno Accademico 2022.
A ZEBRAFISH XENOGRAFT PLATFORM FOR SEMI-QUANTITATIVE ANALYSIS OF INDIVIDUAL STEPS OF THE METASTATIC CASCADE
Metastatic disease represents the main cause of most cancer-related deaths, yet this process remains one of the most enigmatic aspects of cancer biology. Metastatic progression includes multiple distinct phases, such as invasion, intra- and extravasation, seeding at distant sites, micro-metastasis formation and metastatic outgrowth. Only a deep understanding of each of these steps may offer novel targets to be exploited in a clinical setting. Emerging evidence suggests that metastatization is not a genetically selected trait, but the result of phenotypic adaptation to the unfavorable microenvironmental conditions which cancer cells are exposed to (e.g. hypoxia or low nutrients in the primary site or bloodstream). Functional genomic and differential expression screenings allowed to identify numerous potential metastasis genes, that need to be prioritized and validated with appropriate pre-clinical in vivo systems, ideally able to recapitulate and explore the entire metastatic cascade. Mouse models present several critical limitations, including the difficulty to visualize and quantify the metastatic phases, except in sophisticated experimental contexts. Therefore, I propose xenotransplantation of human cancer cells in zebrafish larvae as a system to evaluate primary tumor implantation and growth and to quantitatively dissect metastatic progression, with the purpose of mapping gene functions in the multistep cascade. First, I set up a robust protocol for subcutaneous and intravenous transplantation of fluorescently labeled cells in zebrafish larvae, using the metastatic breast cancer (BC) cell line MDA-MB-231 and non-tumorigenic MCF10A mammary cells as a control. MDA-MB-231 cells survive, proliferate and disseminate in the zebrafish host, developing tumors with conserved histological features and originating extravascular metastases in just 4 days. Using a zebrafish strain with fluorescent vasculature and visually following each fish by live microscopy, I optimized a set of analytical tools to quantitatively investigate incidence, numerosity, size and proliferative index of metastases, as well as measuring frequency of metastasis initiating cells and the severity of metastatic progression. Secondly, I challenged the zebrafish platform by transplanting MDA-MB-231 cells separately interfered for 6 genetic targets, previously identified in our laboratory, as potential novel mediators of BC metastasis. I successfully validated the contribution of 5 out of 6 of these genes in BC metastatization, attributing a specific role in the metastatic cascade for each of them. The identification and in vivo validation of this stage-specific determinants of metastasis offers potential new targets to halt metastatic disease, to be further explored in future pre-clinical and clinical studies, paving the way to novel and effective anti-metastatic therapeutic approaches.
A ZEBRAFISH XENOGRAFT PLATFORM FOR SEMI-QUANTITATIVE ANALYSIS OF INDIVIDUAL STEPS OF THE METASTATIC CASCADE / F. Ruscitto ; tutors: E. Migliaccio, G. Pelicci ; phd coordinator: S. Minucci. Università degli Studi di Milano, 2022 Mar 14. 33. ciclo, Anno Accademico 2021.
EQUILIBRIUM, SYSTEMIC RISK MEASURES AND OPTIMAL TRANSPORT: A CONVEX DUALITY APPROACH
"This Thesis focuses on two main topics. Firstly, we introduce and analyze the novel concept of Systemic Optimal Risk Transfer Equilibrium (SORTE), and we progressively generalize it (i) to a multivariate setup and (ii) to a dynamic (conditional) setting. Additionally we investigate its relation to a recently introduced concept of Systemic Risk Measures (SRM). We present Conditional Systemic Risk Measures and study their properties, dual representation and possible interpretations of the associated allocations as equilibria in the sense of SORTE. On a parallel line of work, we develop a duality for the Entropy Martingale Optimal Transport problem and provide applications to problems of nonlinear pricing-hedging. The mathematical techniques we exploit are mainly borrowed from functional and convex analysis, as well as probability theory. More specifically, apart from a wide range of classical results from functional analysis, we extensively rely on Fenchel-Moreau-Rockafellar type conjugacy results, Minimax Theorems, theory of Orlicz spaces, compactness results in the spirit of Komlós Theorem. At the same time, mathematical results concerning utility maximization theory (existence of optima for primal and dual problems, just to mention an example) and optimal transport theory are widely exploited.The notion of SORTE is inspired by the Bühlmann's classical Equilibrium Risk Exchange (H. Bühlmann, ""The general economic premium principle"", Astin Bulletin, 1984). In both the Bühlmann and the SORTE definition, each agent is behaving rationally by maximizing his/her expected utility given a budget constraint. The two approaches differ by the budget constraints. In Bühlmann's definition the vector that assigns the budget constraint is given a priori. In the SORTE approach, on the contrary, the budget constraint is endogenously determined by solving a systemic utility maximization problem. SORTE gives priority to the systemic aspects of the problem, in order to first optimize the overall systemic performance, rather than to individual rationality. Single agents' preferences are, however, taken into account by the presence of individual optimization problems. The two aspects are simultaneously considered via an optimization problem for a value function given by summation of single agents' utilities. After providing a financial and theoretical justification for this new idea, in this research sufficient general assumptions that guarantee existence, uniqueness, and Pareto optimality of such a SORTE are presented.Once laid the theoretical foundation for the newly introduced SORTE, this Thesis proceeds in extending such a notion to the case when the value function to be optimized has two components, one being the sum of the single agents' utility functions, as in the aforementioned case of SORTE, the other consisting of a truly systemic component. This marks the progress from SORTE to Multivariate Systemic Optimal Risk Transfer Equilibrium (mSORTE). Technically, the extension of SORTE to the new setup requires developing a theory for multivariate utility functions and selecting at the same time a suitable framework for the duality theory. Conceptually, this more general setting allows us to introduce and study a Nash Equilibrium property of the optimizers. Existence, uniqueness, Pareto optimality and the Nash Equilibrium property of the newly defined mSORTE are proved in this Thesis. Additionally, it is shown how mSORTE is in fact a proper generalization, and covers both from the conceptual and the mathematical point of view the notion of SORTE.Proceeding further in the analysis, the relations between the concepts of mSORTE and SRM are investigated in this work. The notion of SRM we start from was introduced in the papers ""A unified approach to systemic risk measures via acceptance sets"" (Math. Finance, 2019) and ""On fairness of systemic risk measures"" (Finance Stoch., 2020) by F. B"
EQUILIBRIUM, SYSTEMIC RISK MEASURES AND OPTIMAL TRANSPORT: A CONVEX DUALITY APPROACH / A. Doldi ; relatore: M. Frittelli ; coordinatore: V. Mastropietro. Dipartimento di Matematica Federigo Enriques, 2021 Feb 26. 33. ciclo, Anno Accademico 2020. [10.13130/doldi-alessandro_phd2021-02-26].
THREE ESSAYS ON POLITICAL ECONOMY OF INSTITUTIONS AND INNOVATION: EVIDENCE FROM SUB-SAHARAN AFRICAN ECONOMIES
This thesis considers the question of how institutions (i.e. political and governance), and innovation affect economic performance in sub-Saharan African economies– a context in which democratic and governance institutions are underdeveloped. To address this fundamental question this dissertation draws on the theories of institutions by North, D. C.(1990), and Acemoglu, D., & Johnson, S.(2005), and the indigenous growth theories by Romer(1986), Lucas(1988), and Schumpeter(1934). To this effect, three different but related in depth investigations are conducted using a panel of sub-Saharan African economies. Using data for a sample of 35 sub-Saharan African economies for 1995–2015 the first article, examines the extent to which political institutions identified as belonging to democratic or autocratic regimes explains the existing differences in innovation across sub-Saharan Africa. While the very few existing studies focus only on the direct effect of institutions, this article examines the impact of the interaction between different regime types and human capital development on innovation in developing countries. The evidence provides very strong support for the direct effect of democratic development on innovation as well as for its indirect effect via its impact on human capital development. However, the results do not support theories that argue in favour of interaction between democracy and human capital, thereby pointing to the need for better calibration of the numerous existing theories and related empirical measures. The second article examines the effect of quality of governance institutions on innovation in 37 sub-Saharan African countries for the period 1996–2016. The empirical analysis followed the ordinary least square and general methods of moment’s regression technique. The motivation for using general methods of moment’s estimation technique is to provide special focus to the issue of endogeneity by estimating general methods of moment’s model. The following general findings are presented. First, governance quality does, in fact, appear to promote innovativeness. Second, for all governance indicators, the effect of the quality of governance institutions follows two channels: directly and indirectly through its positive impact on human capital development. The empirical findings suggest that countries with better quality of governance infrastructure are able to promote innovation in better ways. That is, the results do support theories that argue in favour of the development of governance quality and the improvement of human capital infrastructure to foster national innovation system. These results are found to be robust across alternative empirical specifications tested. Based on empirical panel data for a sample of 37 sub-Saharan African economies for 1996–2016 the third article, investigates the extent to which institutional quality explains the existing cross-country difference in economic performance in sub-Saharan Africa. While most of the existing studies focus only on the direct effect of institutional quality, this article investigates the direct and indirect effects of institutions. It also reflects on impact of the interaction between institutional quality and innovation on economic growth in developing countries. The evidence provides very strong support for the direct effect of institutional quality development on economic performance as well as for its indirect effect via its impact on innovation. However, the results do not support theories that argue in favour of interaction between institutional quality such as democracy, governance quality and innovation, thereby pointing to the need for better calibration of the numerous existing theoretical postulations and related empirical measures. A final epilogue provides explanation on how some of the key trends emerging from the three empirical studies are contributing to both cont
THREE ESSAYS ON POLITICAL ECONOMY OF INSTITUTIONS AND INNOVATION: EVIDENCE FROM SUB-SAHARAN AFRICAN ECONOMIES / D.m. Bekana ; coordinatore: M. Jessoula ; supervisor: G. M. Carbone. Università degli Studi di Milano, 2020 Jun 12. 32. ciclo, Anno Accademico 2019. [10.13130/bekana-dejene-mamo_phd2020-06-12].
STUDY ON HISTONE DEACETYLASE 3 AND ITS PHARMACOLOGICAL REGULATION IN THE PATHOPHYSIOLOGY OF ADIPOSE TISSUE
Introduction: Obesity is a chronic complex disease that poses the risk of developing a plethora of serious health complications (e.g., type 2 diabetes, cardiovascular and cerebrovascular disease, and cancers). Browning of white adipose tissue (WAT) can drain excess nutrients via thermogenesis and thus can be used as a strategy to tackle obesity. Epigenetics and metabolism are closely intertwined in both physiological and pathological conditions. Histone deacetylases (HDACs) are epigenome modifiers regulating adipocyte metabolism. Our previous studies with chemical inhibitors or genetic alterations of HDAC3 have shown metabolic rewiring of white adipocytes towards browning via the activation of a futile cycle of lipolysis and lipogenesis. Obesity is characterized by chronic low-grade inflammation due to secretion of pro-inflammatory cytokines by immune cells, but recent studies demonstrated that adipocyte-derived inflammation is necessary for healthy adipose tissue expansion. Ferroptosis is an iron-dependent programmed cell death characterized by the accumulation of lipid peroxides and may have a role in pathophysiology of adipose tissue.Aim: This thesis is focused on the role of histone deacetylase 3 (HDAC3) in WAT browning and obesity with the following aims: 1) assess the effectiveness of HDAC3 knockdown in precursors and mature adipocytes to i) confirm that HDAC3 is the key HDAC that directs adipocyte fate and to ii) evaluate the window of opportunity for this treatment; 2) evaluate the effects of the peculiar metabolic profile observed in HDAC3 KO mice on adipocyte health, viability, and immunophenotype of adipose tissue; 3) study the effects of LA9498, a novel selective inhibitor of HDAC3, in comparison to MS275, a class I HDAC inhibitor which has already been characterized by our group, to evaluate the efficacy and the optimal effective concentration required to achieve browning of white adipocytes.Results: 1) Hdac3 silencing at early stages of differentiation of C3H/10T1/2 adipocytes promoted a metabolic rewiring of the phenotype towards browning. In particular, adipocyte differentiation and functionality, oxidative metabolism, browning, and mitochondrial functionality increased. In addition, adipocyte pro-inflammatory genes were induced by early silencing of Hdac3. On the other hand, silencing Hdac3 in mature adipocytes did not affect cell phenotype. 2) Ablation of HDAC3 in adipocytes determined the increase in total number of macrophages resident in the epiWAT, but this increase was mostly contributed by the M2 subpopulation. Moreover, Hdac3fatKO mice displayed increased proliferation and differentiation of adipocyte progenitors. FACS analysis confirmed increased adipocytes proliferation and macrophage activation following HDAC3 depletion with RNAi in cultured adipocytes. Alongside new adipocytes, we observed dead adipocytes surrounded by macrophages, suggesting WAT renewal. Integrated analysis of omics data and qPCR analysis revealed ferroptosis as one of the upregulated pathways in WAT of Hdac3fatKO mice and in adipocytes upon silencing of HDAC3 with shRNA adenovector; thus, ferroptosis may be a relevant mechanism triggered in Hdac3fatKO mice to remove metabolically exhausted adipocytes. 3) The HDAC3-selective inhibitor LA9498 is more potent than MS275 and causes upregulation of genes important in i) adipocyte differentiation and functionality, ii) lipid mobilization and β-oxidation, and iii) browning and mitochondrial functionality. LDH test did not reveal toxicity of the compounds at the tested concentrations. Furthermore, LA9498 increased lipid accumulation and parameters of mitochondrial functionality (i.e., basal respiration, ATP production, proton leak, maximal respiration, spare respiratory capacity and non-mitochondrial oxygen consumption).Conclusions: The early epigenetic events mediated by HDAC3 silencing are crucial to commit adipocyte precursors tow
STUDY ON HISTONE DEACETYLASE 3 AND ITS PHARMACOLOGICAL REGULATION IN THE PATHOPHYSIOLOGY OF ADIPOSE TISSUE / L. Coppi ; tutor: M. Crestani ; co-tutor: A. Ferrari ; coordinatore: G. D. Norata. - Dipartimento di Scienze Farmacologiche e Biomolecolari. Dipartimento di Scienze Farmacologiche e Biomolecolari, 2024 Jan 18. 36. ciclo, Anno Accademico 2023.
NEURAL PRECURSOR CELLS AS A POTENTIAL THERAPEUTIC APPROACH FOR RETT SYNDROME: IDENTIFICATION OF THE INVOLVED MOLECULAR MECHANISMS
Rett syndrome (RTT) is a rare, progressive, and profoundly debilitating neurodevelopmental disorder (NDD) and represents the leading cause of severe intellectual disability in females, with an estimated incidence of 1 in 10,000 live births. More than 95% of RTT cases result from sporadic mutations in the MECP2 gene, which is located on the X chromosome (Xq28) and encodes the methyl-CpG binding protein 2 (MeCP2), a nuclear protein that functions as an epigenetic regulator. Affected girls initially exhibit apparently normal development until 6 - 18 months of age, when they enter a severe regression phase characterized by the loss of previously acquired cognitive and motor skills. RTT is an extremely crippling disorder with significant phenotypic variability, influenced by the specific genetic variants causing the disease and by the degree of X chromosome inactivation (XCI). Common symptoms include communication impairments, stereotypical hand movements, respiratory abnormalities, and seizures. At the cellular level, RTT is associated with defects in neuronal morphology, activity, and synaptic transmission.Since the landmark discovery of RTT’s potential reversibility in 2007, significant progress has been made in elucidating the functional role of MECP2 and the consequences of its deficiency. Despite major advances over the past decades, the precise mechanisms underlying RTT pathophysiology remain poorly defined, and a definitive cure remains elusive. Indeed, currently, affected individuals receive symptomatic treatments. However, a significant milestone was reached in 2023 when the U.S. Food and Drug Administration (FDA) approved Trofinetide (DayBue) as the first treatment specifically designed for RTT.Despite this breakthrough, there remains an urgent need for additional therapeutic approaches to improve the severely compromised quality of life of affected individuals. This study was conducted within the broader context of identifying innovative therapeutic strategies for RTT and focused on investigating the potential of neural precursor cell (NPC) transplantation as a promising therapeutic approach for RTT. NPCs are multipotent stem cells that persist in the adult brain. Their therapeutic potential has been extensively explored in both preclinical and clinical studies of neurodegenerative diseases, where they have been shown to be safe and to exert beneficial effects primarily through “bystander” mechanisms. In the context of NDDs, we previously demonstrated that NPC transplantation in Mecp2 knockout (KO) mice rescued the characteristic cognitive and motor deficits in treated animals.When analyzing the transcriptomic profile of NPC-treated mice, we revealed an activation of the IFNγ pathway, that we further validated by observing symptoms improvements after direct IFNγ administrations. These preliminary results suggested a pivotal role for this cytokine in the therapeutic effects of NPCs. Building upon these findings, this study further explored the mechanisms underlying NPC-mediated benefits and leveraged NPCs as a platform to identify new molecular targets for RTT treatment.Using an in vitro transwell co-culture system, we demonstrated that NPC-secreted factors (NPC-SF) specifically rescued synaptic defects in Mecp2 KO neurons without affecting wild-type neurons. Furthermore, we characterized the temporal dynamics of NPC-neuron interactions. Notably, we found that NPCs respond specifically to the RTT pathological environment by releasing factors that promote synaptic repair in Mecp2 KO neurons.Remarkably, we demonstrated that Mecp2 KO NPCs exhibited therapeutic efficacy comparable to the one of wild-type NPCs, paving the way for patient-derived isogenic transplantation strategies. Furthermore, we investigated the role of IFNγ in our in vitro system and confirmed its involvement in NPC-mediated beneficial effects, particularly in the presynaptic compartment.To fu
NEURAL PRECURSOR CELLS AS A POTENTIAL THERAPEUTIC APPROACH FOR RETT SYNDROME: IDENTIFICATION OF THE INVOLVED MOLECULAR MECHANISMS / M. Balbontin Arenas ; tutor: N. Lansdberger ; co-tutor: A. Frasca ; director: N. Landsberger. Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, 2025 Apr 29. 37. ciclo, Anno Accademico 2023/2024.
QUANTUM OBSERVABLES OF OPEN-SHELL SYSTEMS. A THEORETICAL STUDY
This PhD thesis reports original research results concerning the development of theoretical models and computational protocols for the quantification and analysis of two of the most important quantum observables of open-shell systems: the electron spin density and the phosphorescence energy gap. In the first part, a comprehensive theory of the electron spin density topologyis proposed for the first time [1]. Several new notions (spin density critical points, molecular spin graphs, spin density basins, spin maxima and spin minima joining paths etc.) and descriptors (local and integral spin polarization indeces, basin average spin density etc.) are introduced. This analysis reveals that the spin density topology, based on precise mathematical notions, can unveil precious information on the physical structure of spin-polarized systems. In particular, it enables to describe and quantify spin polarization and delocalization mechanisms and, at the same time, to evaluate the dependence of spin density distributions on the adopted level of theory. In the second part instead, the performance of the domain-based local pair natural orbital (DLPNO) variant of the “gold standard” CCSD(T) method for the prediction of phosphorescence energies of aromatic chromophores is investigated for the first time [2]. An extensive analysis of both accuracy and computational cost of the main parameters of the method (basis set, triples correction approximation, dimension of PNOs space) is conducted. Two procedures, the Gold DLPNO-CCSD(T) aimed at maximizing the accuracy and the Silver DLPNO-CCSD(T) aimed at minimizing the computational cost, which result in an excellent agreement with experimental data, are proposed.1. G. Bruno, G. Macetti, L. Lo Presti and C. Gatti, “Spin Density Topology,” Molecules, 25, 3537, 2020.2. G. Bruno, B. de Souza, F. Neese, and G. Bistoni, “Can domain-based local pair natural orbitals approaches accurately predict phosphorescence energies?,” Phys. Chem. Chem. Phys., 24, 14228–14241, 2022.
QUANTUM OBSERVABLES OF OPEN-SHELL SYSTEMS. A THEORETICAL STUDY / G. Bruno ; Tutor: L. Lo Presti, C. Gatti ; coordinatore: D. Passarella. Dipartimento di Chimica, 2022 Sep 16. 34. ciclo, Anno Accademico 2021.
STRUCTURAL ANALYSES OF TRANSCRIPTION FACTOR/DNA COMPLEXES: NFI-X AND NF-Y
Binding of transcription factors (TFs) to discrete sequences in gene promoters and enhancers is crucial to the process by which genetic information is transferred to biological functions. TF structural analysis is the key to understanding their DNA-binding mode and for the design of specific inhibitors. In this context, the present PhD project focuses on two TFs: (1) NFI-X, a TF with unknown structure that binds to the palindromic motif TTGGC(n5)GCCAA and plays an essential role in skeletal muscle development; and (2) NF-Y, a histone-like TF that binds the CCAAT box in promoters of cell cycle genes.(1) The crystal structure of NFI-X was obtained in our lab a few years ago. However, no structural information regarding the DNA-binding mode of NFI-X was available until now. We were able to obtain a functional truncated DNA-binding domain (DBD) construct of NFI-X and purify it for cryo-EM analysis. This allowed us to solve a cryo-EM structure at 3.86 Å resolution, revealing for the first time the architecture of the NFI-X/DNA complex. Surprisingly, the NFI-X/DNA sample formed filaments in the grid after vitrification, which allowed to solve the structure by helical processing. This structure provides critical insights into how NFI-X recognizes its DNA target and mediates dimerization.(2) A new crystal structure of an extended NF-Y construct, termed the NF-Y long-minimal domain (NF-Yl-md), were obtained, which included additional regions of the NF-YC subunit not present in previous structures. These additional regions could potentially influence the DNA-binding and cooperative interactions of NF-Y with other TFs. In parallel, two cryo-EM structures of NF-Y in complex with ATF6 and two cryo-EM maps of NF-Y in complex with USF1 have been elucidated, demonstrating how NF-Y cooperates with these transcription factors to recognize composite DNA elements. These structures provide unprecedented insights into the cooperative assembly of multi-TF complexes on DNA.These findings contribute to the broader understanding of TF-DNA interactions, offering potential avenues for the design of targeted inhibitors that could modulate TF activity in various biological processes.
STRUCTURAL ANALYSES OF TRANSCRIPTION FACTOR/DNA COMPLEXES: NFI-X AND NF-Y / M. Tiberi ; tutor: M. Nardini. - Università degli studi di Milano, Dipartimento di Bioscienze. Dipartimento di Bioscienze, 2025 Jul 15. 37. ciclo
Abstract: Chapter oneThe aim of this paper is to examine the role of monetary and fiscal policies in explainingmacroeconomic fluctuations in Ethiopia using a structural VAR approach, over the period1997/1998:1 to 2016/17:4. Its mains results can be presented as follows: first, an increase ingovernment spending has an expansionary effect on output, while an increase in tax revenuehas a contractionary effect, with spending multipliers larger than net tax revenue multipliers;second, contractionary monetary policy is associated with a fall in output; third, monetary policycontributes to very small fluctuations in output and it is one of the responsible sources of thehigh and persistence inflation in the country; fourth, the contributions of fiscal policy shocks arelarger than that of monetary policy shocks in explaining movements in output, with roughlyequivalent contributions coming from shocks in fiscal policy components. Furthermore, theeffects of fiscal and monetary policy shocks on output and inflation have improved qualitativelyand quantitatively when both policy variables are jointly examined than estimating a separatemodel; suggesting the role of a joint analysis of fiscal and monetary policy shocks.
ESSAYS IN MACROECONOMICS / A.l. Hawitibo ; COORDINATORS: A. MISSALE, E. ROSSI ; SUPERVISOR: E. BACCHIOCCHI. Università degli Studi di Milano, 2020 Feb 25. 32. ciclo, Anno Accademico 2019. [10.13130/hawitibo-alemu-lambamo_phd2020-02-25].
CHARACTERIZATION AND ENGINEERING OF QUANTUM SYSTEMS
Characterization and engineering are the basis for any technological development that aspires to exploit scientific breakthroughs and novel physical phenomena. Nowadays, it is impossible to ignore the exceptional results and developments that quantum mechanics, in combination with information theory, has developed in recent decades. This thesis is part of this line of research, with the scope of providing new results regarding quantum sensing and quantum control. Firstly, the need for quantum systems to work in well-established temperature regimes has brought much attention to quantum thermometry. Establishing new methods to probe temperatures of fragile systems is critical for the development of trustworthy quantum devices. Decoherence is detrimental for many tasks, but the paradigm of quantum probing offers a new perspective on it. Instead of considering it as an undesirable phenomenon, we exploit this inherent fragility to probe features of the environment. Secondly, the ability to jointly estimate parameters is of pivotal importance in several fields, from imaging to gravitational wave detectors. The measurement of quantum systems has several limitations: these might come from an external noisy environment, or they might be quantum in nature, such as the impossibility of simultaneously measuring non-commutative observables. The fundamental understanding of quantum incompatibility in metrology can be impactful both at the foundational and applicative levels. We provide results regarding the quantification of measurement incompatibility, while also studying probe incompatibility, optimizing the initial preparation for the characterization of non-linearities in quantum optical devices. Finally, the last contribution of this PhD thesis consists of a new proposal for quantum search with continuous-time quantum walks. The proposed protocol combines three different fields of research, i.e. continuous measurement, feedback and quantum walks, to provide a novel method to perform quantum search protocols on graphs.
CHARACTERIZATION AND ENGINEERING OF QUANTUM SYSTEMS / A. Candeloro ; tutor: M. PARIS ; coordinatore: M. PARIS. Dipartimento di Fisica Aldo Pontremoli, 2022 Dec 22. 35. ciclo, Anno Accademico 2022.
THE OFD1 PROTEIN CONTROLS AUTOPHAGOSOME BIOGENESIS THROUGH SELECTIVE AUTOPHAGY
Cilia are cellular projections that serve a wide variety of essential functions in mammals. Defects in cilia structure or function have emerged as etiological mechanisms underpinning human diseases called ciliopathies. The OFD1 gene, defective in a rare developmental ciliopathy known as Oral facial digital syndrome type I, encodes for a centrosomal/basal body protein required for cilia formation. Recent data link ciliary structures to autophagy, the major intracellular degradation system, although the mechanisms and the main players underlying this connection are still to be determined.Autophagy is an evolutionarily conserved and strictly regulated lysosomal pathway which plays a wide variety of physiological and pathophysiological roles in cellular homeostasis. Either too little or too much autophagy may contribute to pathological conditions.In the past decade a great deal of progress has been made in the molecular dissection of stimulatory autophagy inputs. On the other hand, our understanding of the mechanisms that restrain autophagy is only partial and far from being complete. Data obtained during my PhD program contribute to the description of a new negative feedback mechanism that inhibits autophagosome biogenesis through selective autophagy-mediated degradation of ATG13, a component of the ULK1 autophagy initiation complex. I demonstrate that the ciliary OFD1 protein is involved in selective autophagy and acts as autophagy receptor for ATG13 via direct interaction with the Atg8/LC3/GABARAP family of proteins. Preliminary data also indicate that excessive autophagy may contribute to the disease pathogenesis. My results add a new tile to the puzzle of the cilia/autophagy interconnection and will help shedding light on these complex biological processes.
THE OFD1 PROTEIN CONTROLS AUTOPHAGOSOME BIOGENESIS THROUGH SELECTIVE AUTOPHAGY / S. Brillante ; added supervisor: M. Morleo ; supervisor: B. Franco. Universita' degli Studi di MILANO, 2020 Dec 11. 32. ciclo, Anno Accademico 2020. [10.13130/brillante-simona_phd2020-12-11].
WATER ECONOMY: SUSTAINABILITY IN BOTTLED WATER SUPPLY CHAIN. THE ROLE OF CONSUMER AND ENTERPRISE TO ENHANCE SUSTAINABILITY IN THE FOOD SYSTEM.
Water is the planet’s most precious natural resource, essential for all life forms and economic activities. Despite covering 70% of the Earth's surface, freshwater represents only 2.5% of total water resources, with just 1% readily available for human use. Population growth, shifting consumption patterns, and climate change are intensifying pressures on this finite resource. This thesis examines environmental sustainability in the water economy, with a particular focus on the bottled water sector, before expanding the analysis to broader sustainability challenges in food consumption.The first part of the research explores sustainability within the bottled water industry, using Italy—one of the world’s largest consumers of bottled water—as a case study. The study identifies a gap in understanding sustainability across the production side of the bottled water value chain. Through an analysis of industry practices, including eco-design, recycled and bio-based materials, and logistics improvements, the research highlights how enterprise-led innovations can contribute to systemic change. Supported by insights from Sanpellegrino, the findings indicate that companies are increasingly adopting sustainable solutions, with European regulatory frameworks playing a crucial role in shaping industry practices. The role of consumer behavior is also considered, emphasizing the importance of environmental labels and certifications in promoting transparency and informed choices.Building on this sector-specific analysis, the research extends to the sustainability of food consumption, focusing on the Water Footprint (WF) of dietary choices in Italy. Using real consumption data from 2005 to 2021, the study reveals a shift away from the traditional Mediterranean Diet (MD), with an overconsumption of red meat and lower vegetable intake contributing to an elevated WF. Through a Sustainable Diet Model (SDM), the research demonstrates that adopting alternative dietary patterns—such as the Atlantic Diet or plant-based diets—can significantly reduce WF. The findings, published in Renewable Agriculture and Food Systems (2024), contribute to the debate on how dietary choices impact water sustainability.The thesis further broadens its scope by assessing the Carbon Footprint (CF) and Ecological Footprint (EF) of Italian food consumption. The analysis confirms that deviations from the MD amplify environmental pressures. Using the SDM, the study evaluates alternative dietary scenarios, showing that flexitarian, pesco-vegetarian, and predominantly plant-based diets significantly lower CF and EF. An unexpected yet significant finding highlights the economic dimension of dietary shifts: sustainable diets, including MD-aligned and plant-based patterns, are more affordable than the current Italian diet. This challenges the misconception that environmentally sustainable diets are costly and positions affordability as a key driver for dietary change. These results were published in Ecological Economics (2023).Finally, the research explores the alignment between supply and demand in promoting sustainability. A study conducted at Université Grenoble-Alpes examines the impact of educational campaigns—Nutri-Score labeling and plant-based meal promotions—on employee food choices in collective catering. While these interventions improved awareness, behavioral shifts remained limited due to entrenched consumption habits and supply constraints. The findings underscore the necessity of coordinated efforts between consumer awareness and structural supply-side changes to drive sustainable dietary transitions.Overall, this thesis provides a comprehensive examination of sustainability in the bottled water and food sectors, highlighting the interconnectedness of production and consumption. By integrating industry practices, policy frameworks, and consumer behavior insights, the research contributes to ongoing di
WATER ECONOMY: SUSTAINABILITY IN BOTTLED WATER SUPPLY CHAIN. THE ROLE OF CONSUMER AND ENTERPRISE TO ENHANCE SUSTAINABILITY IN THE FOOD SYSTEM / E.n. Frola ; tutor: F. D'Alessandro; co-tutor: A. Banterle. - Milano. Dipartimento di Scienze e Politiche Ambientali, 2025 Mar 28. 37. ciclo, Anno Accademico 2024/2025.
INFLUENCE OF THE INTERACTION SCION X ROOTSTOCK ON GRAPEVINE RESPONSE TO DROUGHT STRESS AS A TOOL IN NEW GRAPEVINE ROOTSTOCKS BREEDING PROGRAMS.
Viticulture is expected to face several challenges related to current and future climate changes. Particularly, the vine-growing areas experiencing severe drought stress are expected to increase in the coming years. This could affect grape quality and yield, as well as the character and style of wines, and, more generally, the viticultural potential of many regions. Many studies suggest adaptive strategies to address changing environmental conditions, and the use of drought-tolerant rootstocks is considered an effective mid-term solution to mitigate and manage these negative impacts. However, the range of available rootstocks is limited, and most of them were selected in the late 19th century, when grapevine cultivation practices vastly differed from those of today. As a result, there is a growing demand for drought-tolerant rootstocks that are suitable for modern viticulture. In response to this request, some new genotypes have been released in recent years. Among those, the M-series by University of Milano has shown promising performances in different viticultural contexts.A deep insight into the behaviour of these new rootstocks under water stress conditions, especially in comparison to traditional ones, could provide valuable information, useful at different levels. On one hand, a better understanding of rootstocks performance under drought stress would support winegrowers in making informed decisions when planning new vineyards. On the other hand, it would provide breeders with crucial data on the grapevine responses to water stress, with particular regard to the complex interaction between scion and rootstock, allowing them to better direct their research efforts. To address these questions, three experiments were conducted. The first study evaluates the scion x rootstock interaction effect on drought tolerance, considering two grapevine varieties characterized by different physiological strategies against water stress. More in detail, the study aimed to: (a) analyse the behaviour of ungrafted rootstocks (1103P, M4) and own-rooted vines (Sangiovese, Montepulciano) under increasing water deficit conditions and after rewatering; (b) evaluate the behaviour of the four grafting combinations in the same experimental conditions; and (c) assess the response of the four grafting combinations to different drought dynamics. The second experiment delves into the specific water stress tolerance strategies employed by the two grafting combinations of Pinot Blanc with 1103P and M4, focusing particularly on changes in morphology of the scion, including leaf, petiole and xylem vessels induced by rootstocks. The third experiment examines the influence of rootstock on grapevine production and quality. When selecting a rootstock, it is important not only to assess its tolerance to water stress but also to consider its impact on grape quality, taking into account environmental conditions and oenological objectives. This study analysed six different grafting combinations of Chardonnay with 1103P, SO4, M1, M2, M3, M4, all cultivated in the same vineyard over a four-year period. Data on production per plant, vine vigour, and must quality (sugar content, pH, total acidity, K⁺ concentration) were collected. Furthermore, micro-vinifications of the six combinations were produced in two years and wines sensorial characteristics were evaluated.The results generally highlight the crucial role of the interplay between rootstock, scion, and water stress intensity in shaping drought response mechanisms. Specifically, differences in leaf gasexchange are primarily driven by the variety under moderate water stress and by the rootstock under severe water stress and after rewatering. In particular, M4 outperforms 1103P, demonstrating a superior response under severe water stress conditions and a faster recovery following rehydration. This pattern is consistently observed across all three grafted combina
INFLUENCE OF THE INTERACTION SCION X ROOTSTOCK ON GRAPEVINE RESPONSE TO DROUGHT STRESS AS A TOOL IN NEW GRAPEVINE ROOTSTOCKS BREEDING PROGRAMS / D. Modina ; tutor: L. Brancadoro ; co-tutor: G. Cola ; coordinatore: S. Pilu. Dipartimento di Scienze Agrarie e Ambientali - Produzione, Territorio, Agroenergia, 2025 Mar 21. 37. ciclo, Anno Accademico 2023/2024.
THE INDETERMINATE PRESENT: AN ESSAY ON QUANTUM MECHANICS AND THE OPEN FUTURE
The dissertation is a defense of the following conditional claim: if there areobjective collapses of the wavefunction, then the future is genuinely open.Although this is no radically new idea, the strategy I shall use to defend it isa new one. It proceeds in two main steps. First, building upon the recentliterature on metaphysical indeterminacy in quantum mechanics, I arguefor the view that systems in superposition have be interpreted as objectivelyindeterminate state of affairs. Second, I propose an alternative way to thinkof openness, according to which the future is open as of t, if and onlyif there is an indeterminate state of affair S at t, and S becomes determinateat t’ (with t’ later than t). To argue for the second step, I will give ananalysis of the objective collapses of the wavefunction as the becomingdeterminate of previously indeterminate systems. Furthermore, indeveloping my arguments, I will also make some remarks concerning theontology of objective collapse interpretations of quantum mechanics, theissue of whether metaphysical indeterminacy can be at some derivate levelof reality, and the possibility of the openness of the future being anemergent phenomenon.
"THE INDETERMINATE PRESENT: AN ESSAY ON QUANTUM MECHANICS AND THE OPEN FUTURE / C. Mariani ; tutor: G. Torrengo, Tallant Jonathan, Calosi Claudio. DIPARTIMENTO DI FILOSOFIA ""PIERO MARTINETTI"", 2020 Jan 27. 32. ciclo, Anno Accademico 2019. [10.13130/mariani-cristian_phd2020-01-27]."
DATA DRIVEN ALGORITHMS AND DISTRIBUTED COMPUTING FOR AUTOMATIC MIP DECOMPOSITIONS
Motivated by its implications in the development of general purpose solvers, we address a fundamental research question, that is how to exploit Dantzig-Wolfe decomposition methods for generic Mixed Integer Programs (MIP). In the first part of the thesis, we research how to obtain such decompositions in an automatic fashion. We investigate, with machine learning tools, the link between static properties and good decomposition patterns, highlighting interesting structures. Then, we propose a fully data driven detector, capable of producing good decompositions, and a data driven local search algorithm to reformulate and enhance them. We develop a computational effective framework and test it on unseen MIPs, obtaining better results than state-of-the-art detectors.In the second part of the thesis, we design a concurrent computing framework, which is able to exploit massive parallelism when decompositions allow so. We develop asynchronous and distributed column generation algorithms. Our approach turns out to be, on average, one order of magnitude faster than state-of-the-art solvers.
DATA DRIVEN ALGORITHMS AND DISTRIBUTED COMPUTING FOR AUTOMATIC MIP DECOMPOSITIONS / S. Basso ; tutor: A. Ceselli ; coordinator: P. Boldi. Dipartimento di Informatica Giovanni Degli Antoni, 2021 Mar 22. 33. ciclo, Anno Accademico 2020. [10.13130/basso-saverio_phd2021-03-22].
INVOLVEMENT OF OXIDATIVE BALANCE IN THE ETIOLOGY, PATHOPHYSIOLOGY AND TREATMENT OF STRESS-RELATED PSYCHIATRIC DISORDERS
Psychiatric disorders are severe and heterogeneous pathologies affecting an individual’s cognition, emotional regulation and behaviour. In 2019, 1 over 8 people was living with a mental illness, with major depressive disorder (MDD) identified as the leading global cause of disability, and with the spread of COVID-19 there has been a strong increase in the prevalence of MDD, anxiety disorder (AD) and eating disorders (ED). Moreover, the pharmacological treatment of psychopathologies is still largely ineffective and presents several critical aspects, such as the latency of therapeutic effects and the number of non-responder patients. On these bases, it is crucial to study psychiatric disorders to identify new molecular systems involved in their pathophysiology, with the aim to develop novel pharmacological strategies. In this context, to help the diagnosis and treatment of such pathologies, and based on the fact that some features are shared among the different pathologies, the Research Domain Criteria (RDoC) initiative has been launched by the National Institute of Mental Health. Indeed, the aetiology of many different psychiatric disorders relies on the same basis: the interaction between susceptibility genes and environmental stimuli, with the most important one being stress. Of course, it must be considered that not all the subjects exposed to stress will develop a psychopathology, and that not all types of stressors have detrimental effects on the organism. Indeed, acute stressors can be considered a fuel for the brain and body, helping to achieve expected results, however, if the stressors persist for longer periods, are too intense, or the molecular underpinnings of the body response have been primed to a maladaptive response by genetic or environmental backgrounds, the outcome of stress could be pathological. Moreover, although particularly defining for MDD, another common feature among different psychiatric diseases is the symptom of anhedonia, the inability to experience pleasure in situations considered satisfying. These common aspects are exploited by researchers and clinicians to study and understand them. Thus, my PhD project aims to study, at preclinical level, the molecular effects produced by different types of stress. In order to do so, I exploited different types and combination of stressors, such as the Chronic Mild Stress (CMS) model of depression, combined with acute physical (restraint stress) and inflammatory (lipopolysaccharide or LPS) challenges, and the Activity-Based Anorexia (ABA) model of anorexia nervosa, focusing on the involvement of oxidative balance, investigated mainly at mRNA and protein levels in such aspects. The choice of this molecular system relies on the fact that the brain requires a lot of energy -that is a generator of oxidative stress- to support neuronal activity, and on the fact that oxidative stress could act as a bridge between neuroinflammation and the dysregulation in synaptic processes (i.e. neurotransmitter and receptors expression and/or function, signalling pathways), both important players in the neurobiology of psychiatric disorders. Considering the choice of the CMS paradigm, it is important to underline that this model is able to produce two different subpopulations of stressed animals: the vulnerable one, that develops the anhedonic -depressive-like- phenotype and the resilient one that, despite stress exposure, does not, and this has a great translational value and allowed us to study, at molecular level, also the population that is not susceptible to stress exposure.The results of my PhD project revealed that oxidative balance is crucial for the proper stress response. Indeed, stress exposure was able to cause oxidative stress in different areas of the CNS of vulnerable animals, but not of resilient ones. This derangement is accompanied by a malfunctioning of the antioxidant pathway of Nrf2, modulation that is once
INVOLVEMENT OF OXIDATIVE BALANCE IN THE ETIOLOGY, PATHOPHYSIOLOGY AND TREATMENT OF STRESS-RELATED PSYCHIATRIC DISORDERS / V. Spero ; tutor: R. Molteni ; director: N. Landsberger. Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, 2023 Apr 19. 35. ciclo, Anno Accademico 2022.
POPULATION DYNAMICS AND GEOCHEMISTRY OF BENTHIC AND PLANKTONIC FORAMINIFERA TO INTERPRET THE PALEOCEANOGRAPHIC CHANGES ACROSS THE CENOMANIAN-TURONIAN OCEANIC ANOXIC EVENT 2 AT LOW AND HIGH LATITUDES
This PhD research project has investigated benthic and planktonic foraminiferal dynamics across one of the greatest environmental perturbations that occurred throughout the Earth’s history, namely the Cenomanian-Turonian Oceanic Anoxic Event 2 (OAE 2). It is globally marked by a positive δ13C excursion due to the deposition and burial of large amounts of organic matter in the ocean. The exact causes of OAE 2 are still studied but it is understood that it might be due to a combination of different factors including the emplacement of the Large Igneous Provinces (LIPs), fluctuations of the sea-level, changes in ocean circulation. For this study, four stratigraphic sections have been selected to document how global changes impacted at low and high latitudes. The stratigraphic sections are Clot Chevalier in the Vocontian Basin (SE France) and Eastbourne in the Anglo-Paris Basin (SE England) at low latitudes and IODP Sites U1513 and U1516 in the Mentelle Basin (SW Australia) at high latitudes. Taxonomy, quantitative and statistical analysis have been performed on foraminifera to reconstruct paleoenvironmental and paleoceanographic changes across the OAE 2. In addition, boron isotopes analysis on foraminiferal tests of DSDP Site 463 and of IODP Sites U1513 and U1516 have been conducted to obtain information about ocean acidification across the OAE 2. Samples at low and high latitudes show a continuous foraminiferal record across the OAE 2 displaying high adaptability to extreme environmental conditions, except for a distinct low carbonate interval at Sites U1513 and U1516 characterized by the absence of calcareous taxa. Foraminiferal abundance patterns exhibit notable variances between low and high latitudes with relatively low species diversity and dominance of opportunistic taxa at Sites U1513 and U1516 and with moderate abundances and high species diversity at Clot Chevalier and Eastbourne. This study has delineated six intervals across the Cenomanian-Turonian boundary interval, each marked by unique foraminiferal assemblages reflecting distinct environmental conditions. - Below the OAE 2: high latitudes show low diversity with dominance of opportunistic taxa in the water column that identify a weak stratification and warm waters. At the seafloor benthic foraminifera exhibit taxa tolerant of high organic fluxes, indicating high marine productivity preceding the OAE 2. At low latitudes, a stratified water column indicating meso-oligotrophic regimes and low organic fluxes at the seafloor have been observed. A decrease in ΔpH in surface waters supports enhanced volcanic activity prior to OAE 2, but without significant impacts on foraminiferal tests. Lower part of OAE 2: at high latitudes, biostratigraphic and geochemical data still indicate high marine productivity, warm water conditions and an increase in terrigenous input. In contrast, low latitude planktonic foraminiferal assemblages show the initial thermal disruptions of the stratified water column. At Clot Chevalier the seafloor becomes depleted in benthic foraminifera adapted to low organic fluxes, whereas at Eastbourne a sea-level lowstand is observed. Plenus Cold Event (PCE): at high latitudes, the positive shift in Zr/Rb ratio, the exclusive occurrence of Stensioeina truncata, then increase of epi-infaunal taxa and the decrease in radiolarians indicate improved environmental conditions that might coincide with the PCE. By contrast, low latitudes display clear evidence of the PCE with the occurrence of Boreal planktonic foraminifera and of benthic foraminifera associated to more oxygenated and deeper waters. In addition, a repopulation event is observed at Clot Chevalier. Middle part of OAE 2: the thermal maximum at high latitudes exhibits a remarkable lack of calcareous foraminifera and the dominance of radiolarians indic
POPULATION DYNAMICS AND GEOCHEMISTRY OF BENTHIC AND PLANKTONIC FORAMINIFERA TO INTERPRET THE PALEOCEANOGRAPHIC CHANGES ACROSS THE CENOMANIAN-TURONIAN OCEANIC ANOXIC EVENT 2 AT LOW AND HIGH LATITUDES / G. Amaglio ; tutor: M.R. Petrizzo ; PhD coordinator: M.I. Spalla, G. Muttoni. Dipartimento di Scienze della Terra Ardito Desio, 2025 Feb 07. 37. ciclo
MODULATING TFEB ACTIVITY IN THE BIRT-HOGG-DUBE' SYNDROME BY TARGETING THE VACUOLAR ATPASE
The transcription factor EB (TFEB) is a master regulator of lysosome biogenesis and autophagy, and its activity is primarily controlled by the kinase complex mTORC1. In the Birt-Hogg-Dubé (BHD) syndrome, a genetic disease caused by germline mutations in FLCN gene, TFEB was found to be constitutively active and to promote the development of kidney cysts and cancer. Therefore, inhibition of TFEB activity represents a challenging opportunity for the treatment of the BHD syndrome.We performed a high content siRNA screening to identify correctors of TFEB activation in FLCN-KO cells. Interestingly, top hits were genes encoding subunits of the vacuolar ATPase (V-ATPase). The V-ATPase is a proton-pump protein complex responsible for maintaining the low pH of lysosomes, and it has been described as a positive regulator of mTORC1 signaling, although the exact contribute of the V-ATPase to the amino acid sensing has not been fully elucidated. Our results indicate that the V-ATPase inhibition promotes a mTORC1-dependent TFEB cytosolic re-localization in FLCN-KO cells, thus pointing to a novel mechanism of regulation of TFEB activity in BHD cellular models mediated by the V-ATPase. Besides, our data suggest that the Ragulator-Rag GTPases complex plays a crucial role in controlling TFEB subcellular localization, even in absence of FLCN, through its interaction with the V-ATPase.This study aims at dissecting how the V-ATPase regulates mTORC1 signaling and TFEB activity, while searching for compounds able to target this axis, with the purpose to identify novel and effective therapeutic approaches to limit kidney pathologies associated with BHD syndrome.
MODULATING TFEB ACTIVITY IN THE BIRT-HOGG-DUBE' SYNDROME BY TARGETING THE VACUOLAR ATPASE / C. Vilardo ; supervisor: A. Ballabio ; added supervisor: C. Di Malta ; internal advisor: P. Grumati ; external advisor: P. Saftig. Dipartimento di Oncologia ed Emato-Oncologia, 2023 Dec 04. 35. ciclo, Anno Accademico 2023.
INFORMAZIONE NON FINANZIARIA E RESPONSABILITÀ DEGLI AMMINISTRATORI
The thesis is dedicated to non-financial reporting and to the related aspects concerning director’s liability towards shareholders and third parties. Non-financial reporting’s legal framework is essentially provided by a complex of provisions introduced by the European legislator with Directive 2014/95/EU, amending Directive 20103/34/EU, by the transposing legislative decree no. 254 of 2016 and now also by recent Directive 2022/2464/EU. The introduction of the sectoral legislation has foreseen non-financial information as an act of the directors and has made its preparation mandatory with reference to certain large entities, thus giving special emphasis to the emerging practice concerning the disclosure of this type of information.The work is divided along three progressive research lines, corresponding to its three chapters.In the first chapter, before proceeding to the description of the current legislation and the presentation of legislation in progress, we wanted to give account of the context, not only the legal one but also and above all the cultural one, in which non-financial reporting finds its origins. The introduction of non-financial reporting is a normative expression of corporate social responsibility theories, of the integration of environmental, social and governance factors in business policies, of the growing attention to the concept of sustainability. The chapter contains a brief account of this framework also in the intent of assessing non-financial reporting as a tool for the legal embodiment of issues that have in the past been more often object of soft law interventions.The second chapter is dedicated to questioning the legal nature of non-financial reporting, also in order to address with greater awareness its possible implications in terms of liability. The purpose of the second chapter is therefore to arrive to a legal qualification of the act itself, reconducting it to well-known civil categories. The chapter also contains a record of several literal expressions which are in concreto often used by firms whilst drafting non-financial statements. This series of examples, which clearly is not exhaustive, is meant to link the theoretical analysis to practical experience and thus to the ways in which non-financial reporting is carried out by enterprises.The third chapter is entirely dedicated to the possible consequences in terms of liability, arising from the drafting of non-financial reporting, for the directors. The latter are the disclosure’s actual authors and to their responsibility the act is attributed. The possibility that non-financial reporting may conduct to liability for corporate bodies (and/or for the company) stems from the consideration that we are dealing with information, the content of which is still flexible, and which is therefore often not extremely punctual, but nevertheless capable of influencing the shareholders’ and third parties’ choices. The possible liability arising because of false or inaccurate non-financial disclosure can be ascribed to the very large and heterogeneous category of misrepresentation, which has been extensively investigated by our private law doctrine. And the company law tool which appears to be functional for the protection of the recipients of the information who manage to prove they have relied on the truthfulness and correctness of the non-financial statement and on its basis made investing or other different decisions, is article 2395 of the Italian Civil Code. To the latter’s systematic framework the working hypothesis must therefore be brought back.
INFORMAZIONE NON FINANZIARIA E RESPONSABILITÀ DEGLI AMMINISTRATORI / A. Briguglio ; tutor: V. Pinto, R. Sacchi ; coordinatore: G. Ludovico. Dipartimento di Diritto Privato e Storia del Diritto, 2023 May 05. 35. ciclo, Anno Accademico 2022.
PUBERTAL INDUCTION IN GIRLS WITH HYPOGONADISM: ESTROGEN REPLACEMENT THERAPY OUTCOMES AND OPTIMIZATION OF PROGESTERONE INTRODUCTION
Background: Pubertal induction in girls with hypogonadism through estrogen replacement therapy (ERT) aims at mimicking physiological puberty. To date, the best induction regimen, being physiological and effective at the same time, is still to be established. Moreover, available data in the literature show a poor uterine development at the end of the induction in more than a half hypogonadal patients: this probably represents one of the most important contributing factors for negative pregnancy outcomes of this specific population.Aims: We analysed longitudinal data on pubertal induction in girls with hypogonadotropic hypogonadism or primary ovarian insufficiency (congenital, acquired, isolated or associated with multiple pituitary hormone deficiencies, either associated with Turner Syndrome or secondary to late effects of cancer treatment) in order to insight into auxological and uterine outcomes in the light of different underlying diagnosis and regimens used.Methods: Due to the lack of licensed specific hormone preparations for pubertal induction, randomized-controlled trials are not feasible; thus, a multicentre clinical registry was created. Out of 106 hypogonadal girls (chronological age>10.9 years, Tanner stage≤2) who received ERT for pubertal induction, we considered 95 girls (median age 13.5 years) treated with transdermal (TD) 17β-estradiol patches for at least one year (median 3.3 years). Induction was started at a median dose of 0.14 mcg/kg/day TD 17β-estradiol, with a six-monthly increase. Auxological, biochemical (estradiol levels) and radiological (pelvic US) data were collected at baseline and during follow-up. In 61/95 girls, progesterone was introduced after a median of 2.2 years. Induction was considered completed for the 49/95 patients who were started on combined oral contraceptive (COC) or progesterone plus at least 50 mcg/day or 1 mcg/kg/day of TD 17β-estradiol.Results: At the end of induction, 90.0% patients had achieved Tanner stage IV and 41.0% Tanner stage V, the latter being associated with 17β-estradiol dose at progesterone introduction (P=0.034). Uterine longitudinal diameter (ULD) showed a gradual increase during ERT and a significant correlation with 17β-estradiol dosage (P<0.0001) at any point of induction. Nonetheless, the final ULD was>65 mm in only 17/45 (38%). At multiple regression analysis, a history of pelvic irradiation represented the major determinant of reduced final ULD (P=0.034). After correction for uterine irradiation, ULD was associated with 17β-estradiol dose at progesterone introduction (P=0.043). Final ULD was not significantly different from the one assed after COC introduction.Conclusions: Our results show that therapeutic goals should be individualized and tailored to the underlying different diagnostic categories. Moreover, adolescence represents a critical window for uterine and breast development: indeed, given the evidence that progestins may hamper the subsequent changes in uterus volume or the achievement of the last Tanner stage, such therapy should be started only in the presence of a concomitant adequate ERT dose and an appropriate clinical response (uterus and breast development). At present, data collection is ongoing to confirm these assumptions on a larger scale and to identify the best induction regimen for each diagnostic category, with particular focus on the specific needs of childhood cancer survivors.
PUBERTAL INDUCTION IN GIRLS WITH HYPOGONADISM: ESTROGEN REPLACEMENT THERAPY OUTCOMES AND OPTIMIZATION OF PROGESTERONE INTRODUCTION / G. Rodari ; tutor: M. Arosio ; coordinator: M. Del Fabbro. Dipartimento di Scienze Cliniche e di Comunità, 2022 Mar 16. 34. ciclo, Anno Accademico 2021.
OPTIMIZING MICROCLIMATE CONTROL STRATEGIES IN PRECISION AGRICULTURE USING MACHINE LEARNING TECHNIQUES AND IOT SENSORS DATA
The overarching aim of this PhD thesis is to push the boundaries of microclimate monitoring in precision agriculture through the application of machine learning to IoT sensor data. The sensors provide vast amounts of detailed microclimatic information, which can be effectively processed by machine learning algorithms to optimize control strategies for precision agriculture.The thesis aims to advance the state of the art in microclimate control by integrating machine learning techniques with sensor data, focusing on two key dimensions: spatial and temporal.The scientific work presented in this thesis was developed in collaboration with DNDG srl, a company specialized in the development of fully customized, high performance IoT industrial systems business solutions and digital products, and a partner of the multinational Finder Group.From a spatial perspective, this thesis aims to optimize microclimate monitoring in outdoor agricultural fields with a complex terrain conformation, such as vineyards. As will be discussed in the Introduction, microclimate monitoring can be performed using either remote sensing or networks of IoT sensors. While remote sensing provides highly accurate results at continuous spatial scales, it is expensive and does not allow for continuous, real-time monitoring. IoT sensor networks offer a more affordable solution for real-time monitoring of specific locations within a field. However, these measurements are localized and may miss significant microclimatic patterns elsewhere in the field. An ideal solution would involve deploying a large number of sensors, but this would significantly increase maintenance costs. To address this, zones with similar microclimatic patterns need to be identified, allowing for the strategic placement of sensors within each zone.The literature currently lacks a clear strategy to achieve this. Thus, there is a need for a method that can use IoT data to continuously reconstruct microclimatic variations and identify characteristic patterns. Given the mathematical complexity of modelling the physical behaviour in this task, the approach is based on data-driven methods such as neural networks and machine learning clustering algorithms.This thesis tackles the problem in two steps: (1) using neural networks trained on IoT sensor data to obtain a continuous description of microclimate variations at the meter scale across the entire field; and (2) identifying spatial areas with similar microclimate variations using a machine learning clustering algorithm.From a temporal perspective, this thesis proposes the application of neural networks tailored for time series forecasting to enhance time-based monitoring of microclimate effects on precision agriculture systems. The temporal aspect is addressed through two distinct scenarios.First, a case study is presented regarding the use of neural networks to monitor milk production data in a commercial dairy farm equipped with Automatic Milking Systems, with a focus on the influence of microclimate conditions. Milk production is highly sensitive to the microclimatic conditions experienced by dairy cows. To improve production and enhance animal welfare, precision agriculture should implement strategies that forecast milk production based on the relationship between past microclimate conditions and production data. This thesis applies a modern neural network for time series forecasting (Time-Series Mixer) to predict future milk production, demonstrating how AI can be utilized to optimize production for commercial dairy farms.The second scenario focuses on developing a strategy to improve weather forecasting for local agricultural sites, with a particular emphasis on predicting hazardous events that could impact agricultural production. Accurate knowledge of future weather conditions is crucial for optimizing risk management strategies. Numerical weather predictions generate forecasts at global or regional scales, which lack p
OPTIMIZING MICROCLIMATE CONTROL STRATEGIES IN PRECISION AGRICULTURE USING MACHINE LEARNING TECHNIQUES AND IOT SENSORS DATA / M. Zanchi ; supervisore: C.A.M. La Porta ; tutor: P. Di Nunzio ; coordinatore: M. Guarino. Dipartimento di Scienze e Politiche Ambientali, 2025. 37. ciclo, Anno Accademico 2024/2025.
DONT' BE FOOLED BY FAKE NEWS? MAPPING THE SOCIAL, COGNITIVE, AND POLITICAL MECHANISMS OF MISINFORMATION BELIEF.
Why do people believe in fake news? I explore this question using five studies on more than 4200 participants. While there is extensive research on this question, most studies have primarily tried to identify specific individual mechanisms, e.g. cognitive styles, without considering the possible interplay of various mechanisms. Instead, this dissertation has jointly considered five drivers of misinformation beliefs: cognitive styles, motivated reasoning, anti-elitism, institutional trust, and social norms. The second contribution of this doctoral thesis is the inclusion of these latter three explanations (one of which - social norms - is also experimentally manipulated), as the literature often focused on cognitive drivers while only rarely exploring social ones. This dissertation also aimed to make relevant methodological contributions. First, instead of using artificial or semiartificial stimuli to test participants’ truth discernment, it collected and validated a new dataset of 80 existing social media posts. Second, it also tested its hypotheses in Italy, outside the widely explored context of English-speaking countries. The results of these studies indicate that cognitive styles, such as the tendency to rely on intuition or analytical thinking, significantly influence participants’ ability to discern truth, even when using real social media posts as stimuli. However, the effects of other drivers were less pronounced. Motivated reasoning, for instance, had only marginal effects, possibly due to the low perceived partisanship of the collected posts. Similarly, I found no consistently significant effects of anti-elitism and institutional trust, except for trust in universities. Regarding social norms, I observed that individual motivations for information accuracy were associated with higher truth discernment, while social expectations had no discernible impact. The social norm treatment message yielded mixed results but was largely inconclusive. My findings have two main insights: for methodological issues, my thesis testifies to the importance of using more realistic stimuli selection in experimental research on misinformation; for substantive issues, my study shows that cognitive drivers, although crucial, cannot be considered the sole determinants for misinformation beliefs.
Dont' Be Fooled By Fake News? Mapping The Social, Cognitive, And Political Mechanisms Of Misinformation Belief / F. Torreggiani ; tutor: G. Ballarino, A. Szekely ; coordinatore: M. Guerci. - Milano. Dipartimento di Scienze Sociali e Politiche, 2025 Feb 10. 36. ciclo, Anno Accademico 2022/2023.
EPIGENETIC AND FUNCTIONAL ASSESSMENT OF ENHANCEROPATHIES ACROSS HUMAN MODELS: FOCUS ON GABRIELE-DE VRIES SYNDROME
Yin Yang 1 (YY1) is a ubiquitous zinc finger transcription factor (TF) that occupies active enhancers and promoters contributing to physical interactions between these regions via DNA looping. Increasing evidence shows that disruption of non-coding regions such as enhancers is prevalent across different neurodevelopmental disorders (NDDs) with intellectual disability (ID) features. Indeed, YY1 haploinsufficiency causes a NDD with ID, named Gabriele-de Vries syndrome (GADEVS). Although it is known that YY1 controls the expression of a dazzling list of genes and influences various cellular processes in numerous cell types, the impact of this TF in the neurodevelopment of the human cortex is yet to be unraveled. By taking advantage of disease-modeling as a tool to investigate the pathogenesis of GADEVS across different time points and tissues we gathered new insights about how YY1 haploinsufficiency exerts such a dramatic phenotype in individuals carrying mutations. We reprogrammed patient-derived and healthy somatic cells into induced-pluripotent stem cells (iPSCs) and observed, already at the pluripotent stage, a major transcriptional dysregulation. Moreover, since YY1-mutated patients exhibit ID features, we differentiated our cohort of iPSCs into cortical neurons as well organoids and were able to capture stage-specific striking features, not only at the transcriptomic level, but also structural and compartmentalization impairments. Of note, YY1-mutated neurons displayed synaptic disparities, sufficient to induce astrogliosis-like features in surrounding astrocytes, both shown to be critical for proper brain function and plasticity forms in the CNS. Instead, in cortical organoids we recapitulated features of abnormal ventricle formation, pathological hallmarks observed in GADEVS patients and mice models followed by ID and developmental delay. This study showed, for the first time, the molecular signatures that possibly lead to cognitive defects in human patients and provide the first solid foundation for the development of therapeutic strategies and drug screening in the future.
EPIGENETIC AND FUNCTIONAL ASSESSMENT OF ENHANCEROPATHIES ACROSS HUMAN MODELS: FOCUS ON GABRIELE-DE VRIES SYNDROME / M.c. Faria Pereira ; tutor: G. Testa : coordinatore: S. Minucci. Università degli Studi di Milano, 2022 Dec 16. 33. ciclo, Anno Accademico 2021.
MODELLING AND NUMERICAL STUDY OF MARBLE SULPHATION IN CULTURALE HERITAGE: A RANDOM PDE-ODE SYSTEM
The final scope of this thesis is to investigate from a numerical point of view a new approach to the mathematical modelling of the degradation process in the context of Cultural Heritage, focusing on the description of the marble sulphation chemical reaction. Our contribution is a numerical analysis of a novel random model of the sulphation reaction on the half-line. Given a system of non-linear reaction-diffusion partial and ordinary differential equations (PDE-ODE) system, already present in the literature and describing the penetration of sulphur dioxide through the pores of a calcium carbonate-based surface which reacts with a consequent formation of gypsum, a source of randomness has been introduced as a Dirichlet dynamical boundary condition, with the aim to better model the sulphur dioxide evolution in the air. A direct observation of some recent sulphur dioxide time series recorded in Milan suggests that an appropriate model needs to incorporate a significant stochastic fluctuation component, boundedness and a mean-reverting behaviour. The novelty relies on the choice of a class of stochastic processes with such properties, known as Pearson diffusions, and which can be introduced as solutions of an It\^o's stochastic differential equations (SDE). For completeness, positiveness, boundedness, and stability for the Pearson SDE, as well as its numerical scheme, known as Lamperti Sloping Smooth Truncation, based on the well-known Euler-Maruyama method, are discussed and implemented, respectively. The entirely original results are presented in the second part of the thesis. The numerical approximation of the overall system, combination of a non-linear reaction-diffusion PDE-ODE system with a stochastic boundary condition given by a specific Pearson process is investigated. Following recent theoretical results about the existence and uniqueness of a mild solution of this random system, a new numerical splitting scheme is proposed: an autonomous heat equation which inherits the stochastic boundary condition, and a non-linear and non local random system, coupled with the first, but with deterministic boundary conditions. We present an FTCS approximation and we provide the stability conditions for the discrete scheme. Furthermore, the order of spatial accuracy is numerically estimated to be one. Numerical quantitative and qualitative analyses of the impact of the randomness at the boundary are illustrated, both in the framework of single realizations of the process, i.e. in a pathwise sense, and also by considering distribution, first and second moments of large samples. We analyse both slow and accelerated regimes, by considering increasing values of the reaction rates. In the fast reaction case, a formation of a moving stochastic front can be observed.The final part of the thesis presents a theoretical convergence result. We consider a semi-discrete approximation of the system and prove the convergence of the discrete solution to the unique mild solution of the original random system, through a space-discrete approach. The same splitting strategy is adopted to derive some a priori estimates for the splitted discrete variables. Relevant results on compact embedding for time-space Besov spaces on the lattice as well some recent estimate results for the discrete heat kernel are exploited to prove a useful estimate for the solution to the discrete random heat equation. Furthermore, an $L^2$ a priori estimate for the second splitted variable together with its discrete derivative is provided. The final convergence result is obtained by a compactness argument, by first introducing piecewise constant interpolation and discretization operators on the lattice and by exploiting the uniform a priori estimate for the total system variable.
MODELLING AND NUMERICAL STUDY OF MARBLE SULPHATION IN CULTURALE HERITAGE: A RANDOM PDE-ODE SYSTEM / F. Arceci ; tutor: S. Ugolini ; cotutor: D. Morale. Dipartimento di Matematica Federigo Enriques, 2025 May 19. 37. ciclo, Anno Accademico 2023/2024.
LA DOCUMENTAZIONE INTERDITTIVA ANTIMAFIA:ORDINAMENTO SINGOLARE E CORTOCIRCUITI DELLA LEGALITÀ
"The anti-mafia interdiction documnetation: singular order and short-circuits of legality The research proposes to address the relationship between administrative tools for combating organized crime and procedural guarantees of their recipients, looking at the potential friction with the supranational system. Precisely, the reference goes to the ""interdiction antimafia documentation"", in the form both of communication and information. Having analyzed its legal nature, the first research question regards the instrument's compatibility pursuant constitutional, Euro-Unitarian and conventional principles, with particular reference to the principle of legality, in its integrated and multi-level status.Closely related to the former, it is the issue concerning the possibility to extend the judicial interpretation on the ""substantially criminal"" sanctions or to reach a different solution. In this way, the essay leverages the link that binds legal instrument of protection with powers of administrative courts. In order to achieve the expected scientific results, this research follows a path between thematic areas of study placed in logical-argumentative succession. So, starting from a general framing of the administrative tools in countering organized crime, the thesis ends studying the judicial review limits on technical discretion in anti-mafia interdiction orders."
LA DOCUMENTAZIONE INTERDITTIVA ANTIMAFIA:ORDINAMENTO SINGOLARE E CORTOCIRCUITI DELLA LEGALITÀ / L. Filieri ; tutor: P. Provenzano ; coordinatore: F. Basile. Dipartimento di Diritto Pubblico Italiano e Sovranazionale, 2022 Jul 11. 34. ciclo, Anno Accademico 2021.
AM I RADICAL, RIGHT? THE RADICAL RIGHT FAMILY IDEOLOGICAL OUTLOOK AND RADICAL RIGHT ATTITUDE AMONG THE PUBLIC
The emergence of Radical Right Parties (RRPs) has been one of the most studied European political phenomena in the last thirty years, producing a prolific flow of research on both the ideological outlook of the radical right party family and the characteristics of its voters. Nonetheless, the academic debate on these issues is far from being appeased. The following work contributes to the debate by looking at the rise of RRPs in Europe through the perspective of their ideological outlook and by investigating both the supply side and the demand side by means of a data-driven machine learning approach. The literature on RRPs ideological outlook has been prolific and discordant, yet most scholars tend to agree on two main core ideological features that appropriately characterize the radical right family and set it apart from others: nativism and authoritarianism. These core features are complemented by other ancillary elements, on which the debate is more open. The most discussed example is represented by the evolution of RRPs’ positions on economic issues. On this concern, some scholars claim that we are facing a real differentiation within the radical right that needs to be included in the ideological definition of the family, while others argue we are witnessing a series of blurrying processes that intervene on secondary issues holding limited definitory power. As far as radical right voters are concerned, their behavior has been analyzed under many different lenses, still the only condition on which all scholars agree is the centrality of individual attitudes towards immigration in shaping radical right voting. In a few words, we describe the awkward situation where ideologically complex parties, which are not considered single-issue parties, seem to mobilize voters on the basis of a single issue. The aim of the study is to put theoretical studies on the radical right ideology to an empirical test, starting from party-level data and by checking whether those who have been defined as the core features of the radical right family have a crucial role in defining the ideological borders of the radical right or not. Then, we define the potential voting base by following the ideological characterization that emerged empirically. Results confirm the centrality of both nativism and authoritarianism (and Euroscepticism, in addition) in shaping the radical right ideological outlook, while economic positions are the main division line that shapes heterogeneity within the family. The empirical clusters based on the criteria defined previously produce a classification which is coherent with the ones provided by the literature. Results also give empirical evidence of a process of radicalization that characterizes a group of center-right parties, which progressively get close to nativist and authoritarian positions, in continuity with the findings of studies on center-right radicalization. We also confirm that the emergence of a nativist, authoritarian and Eurosceptic set of attitudes among the public, which significantly shapes the probability to vote for a RRP.
AM I RADICAL, RIGHT? THE RADICAL RIGHT FAMILY IDEOLOGICAL OUTLOOK AND RADICAL RIGHT ATTITUDE AMONG THE PUBLIC / F. Piacentini ; tutor: C. Vezzoni ; phd director: M. Jessoula. Dipartimento di Scienze Sociali e Politiche, 2022 Dec 06. 34. ciclo, Anno Accademico 2021.
MEASURING CMB POLARIZATION WITH THE LSPE EXPERIMENT: SIMULATION AND SENSITIVITY STUDIES FOR THE STRIP INSTRUMENT
The search of primordial B-mode fluctuations in the polarization pattern of the Cosmic Microwave Background (CMB) is one of the most active and challenging fields in modern observational cosmology. A detection of primordial B-modes would in fact provide a strong evidence in favour of the Inflationary paradigm, shedding light on the physics of the early Universe.The Large Scale Polarization Explorer is an Italian-led experiment designed to measure the CMB polarization at large angular scales. Its primary goals are the constraint of the B-mode component down to r = 0.03 and the study of the polarized emissions of our Galaxy. LSPE will observe 25% of the sky in the Northern hemisphere relying on the synergy of two diﬀerent instruments, a balloon-borne experiment (SWIPE) and a ground-based telescope (STRIP), complementary for frequency coverage and technology.STRIP, in particular, will observe the 43 GHz sky with an array of 49 polarimeters in order to characterize the Galactic synchrotron signal. STRIP will be installed at the Teide Observatory in Tenerife (Canary Islands, Spain) in mid-late 2021, and at least two years of observations will follow.This PhD thesis describes the Unit Tests campaign of STRIP polarimetric modules, from the test procedures to the data analysis, focusing in particular on the characterization of the bandpass response.A simulation pipeline, developed for the STRIP experiment in order to assess its scientific performance, is also presented. Particular attention is given to the generation of realistic data streams and to the implementation of a map-maker, following the destriping technique.The results of end-to-end simulations of STRIP observation, also exploiting the results of the Unit Tests, are finally presented. In particular, the behaviour of the destriping algorithm in the specific case of the STRIP instrument has been evaluated, together with the impact of some systematics eﬀects.
MEASURING CMB POLARIZATION WITH THE LSPE EXPERIMENT: SIMULATION AND SENSITIVITY STUDIES FOR THE STRIP INSTRUMENT / S. Caprioli ; supervisor: B.Caccianiga, M.Tomasi ; coordinatore: M.Paris. Università degli Studi di Milano, 2020 Feb 19. 32. ciclo, Anno Accademico 2019. [10.13130/caprioli-silvia_phd2020-02-19].
THE MMR VACCINE-AUTISM CONTROVERSY IN THE POST-TRUTH ERA: A CORPUS-ASSISTED DISCOURSE ANALYSIS OF A NEWSPAPER AND FACEBOOK CORPUS
The present dissertation is a corpus-assisted (critical) discourse analysis (Baker 2006; KhosraviNik and Unger 2016) of the UK newspaper coverage of the MMR vaccine-autism “controversy”. This “controversy” concerns the alleged link between the triple MMR vaccine and the onset of autism, that was first suggested in 1998 by Andrew Wakefield and his colleagues in a paper published in the prestigious British medical journal The Lancet. Despite having been subsequently discredited and retracted, and despite the fact that Andrew Wakefield was stripped of his medical license by the British General Medical Council, the paper has arguably contributed greatly to the negative perception of vaccines by the British public: it fostered a climate of scepticism towards experts and authorities, and it effectively created a public debate, re-presented in the mass media, where objective facts and evidence were pitted against emotions and personal beliefs (see, for example: Boyce 2006; Clarke 2008; Stöckl and Smajdor 2017). In this sense, the MMR vaccine-autism debate may also be interpreted considering more recent discussions about a post-truth epistemology (D’Ancona 2017) and the spreading of mis/disinformation and fake news. In order to explore the discursive characteristics of this debate, a corpus of news articles from major UK newspapers was built and analysed; the quantitative analysis was carried out using the corpus analyser AntConc (Anthony 2020), while the qualitative analysis was carried out manually through close reading. The analysis aimed at exploring the way in which proponents and opponents of vaccination used language – and especially evaluation (Bednarek 2006a, 2006b), polyphony (Fløttum and Gjerstad 2013) and argumentative storytelling (Carranza 2015) – to legitimize their views and delegitimize their opponents’. To this end, special attention was paid to editorials and readers’ letters, which were largely ignored by previous studies (Clarke 2008) but are here seen as argumentative texts (Richardson 2007) allowing for reader participation and engagement, where audiences can publicly react to the news and discuss it.The second part of the research is devoted to the analysis of a corpus of users’ comments taken from the Guardian and the Daily Mail Facebook pages, with the aim of carrying out a comparison between online and offline debates about the MMR vaccine. Such a comparison is proposed as one possible way to explore the role played by the internet and social media in re-presenting medical debates and in shaping the so-called “post-truth” society.The results of the analyses point to a marked polyphony in newspaper articles, where multiple social actors are re-presented (chiefly children, parents, medical doctors, and the government), often using largely the same linguistic strategies and frames; thus, different views are equally quoted and legitimised, irrespective of the amount of scientific evidence supporting them. Social actors themselves exploit participatory genres such as letters to the editor and Facebook comments to foster their own views using a variety of linguistic and argumentative means, the most distinctive of which is argumentative storytelling, whereby personal experiences with vaccinations are used as evidence to support one’s positions towards the medical procedure. Moreover, online participants regularly refer to each other using labels such as pro-vaxxer and anti-vaxxer (which, on the other hand, are infrequent in the newspaper corpus) used to summarise one own’s identity and system of beliefs. Thus, trust in vaccines is transformed into a polarising, partisan, and ultimately profoundly ideological issue.
THE MMR VACCINE-AUTISM CONTROVERSY IN THE POST-TRUTH ERA: A CORPUS-ASSISTED DISCOURSE ANALYSIS OF A NEWSPAPER AND FACEBOOK CORPUS / C. Fiammenghi ; tutor: L. Pinnavaia ; coordinatrice del corso di dottorato: M. V. Calvi. Dipartimento di Lingue, Letterature, Culture e Mediazioni, 2022 Apr 06. 34. ciclo, Anno Accademico 2021.
IDENTIFICATION OF NCAM1 AS A NOVEL PROGNOSTIC PROSTATE CANCER STEM CELL BIOMARKER
Serum prostate-specific antigen and Gleason grade are parameters routinely used for risk stratification in prostate cancer (PCa), but they present some limitations in the prediction of disease progression and in their use to guide clinical decision making. Prostate cancer stem cells (PCSCs) are widely considered to be responsible for tumorigenesis, disease progression and therapy failure. Their identification and characterization are mandatory for understanding the intricate intratumoral heterogeneity (spatial and molecular) of PCa and for developing relevant clinical tools to effectively manage patients and tailor therapy. Here, we proposed the surface glycoprotein neural cell adhesion molecule (NCAM1/CD56), a known marker of neuroendocrine (NE) cells, as a novel PCSC marker that provides prognostic information and molecular insights into the process of tumorigenesis. NCAM1 defines clusters of cells enriched in proliferative inflammatory atrophy (PIA) regions, without NE traits (hereafter referred as NCAM1+). In a retrospective cohort of 406 PCa patients treated with radical prostatectomy (RP), we uncovered that NCAM1 is an independent prognostic marker for predicting distant metastasis and biochemical recurrence and its expression in radical prostatectomy biopsies concurred with diagnostic biopsies (concordance 87.6%). Using the human cell lines, LNCaP (androgen-sensitive) and DU145 (androgen-insensitive), we found that NCAM1, but not other candidate PCSC markers, allowed FACS-based prospective purification of PCa cells displaying i) unique self-renewal ability in vitro, in a serial 3D-Matrigel organoid propagation assay, ii) tumorigenic potential upon limiting dilution transplantation in vivo. Relevant to real-life human PCa, we found that the ability to generate primary-derived organoids (PDOs) from dissociated high Gleason PCa biopsies exclusively resided in the purified NCAM1+ cell fraction, a property that was efficiently inhibited by treatment with an anti-NCAM1 blocking monoclonal antibody. We also found that the progressive development of adenocarcinoma in transgenic TRAMP mice crossed with NCAM1-/- mice was blocked at very early stages of tumorigenesis, indicating that genetic NCAM1 ablation prevents premalignant lesions to expand and progress to advanced stages. PCa is a paradigm tumor model for clinical, spatial and molecular heterogeneity and this heterogeneity is reflected in the NCAM1+ cell population. Single cell-RNA sequencing (sc-RNASeq) of purified NCAM1+ cells (cell lines and primary human PCa biopsies) uncovered heterogenous cellular states reflected in several distinct clusters. Phylogenetic tree reconstruction of the evolutionary relationship among the different clusters along with single cell trajectory analysis revealed the existence of a cell fraction with basal traits (p63+/AR-/CD117+ cells) and a quiescent phenotype, sitting at the apex of the hierarchical structure of the NCAM1+ population. These cells were functionally characterized by Hedgehog signaling which drives NCAM1+/CD117+ -PCSC self-renewal ability. Moreover, they were molecularly characterized by a transcriptional signature called “Stem Score” which could have potential as a prognostic tool for identifying patients at risk of biochemical recurrence (BCR) and distant metastasis.Androgen deprivation therapy (ADT) is the standard management for advanced PCa. Despite its initial effectiveness, the majority of patients relapse and develop castration resistant prostate cancer (CRPC), which is thought to be mediated by resistant PCSCs. ADT-treated NCAM1+ cells isolated from both dissociated human PCa biopsies and the LNCaP cell line enter into a quiescent state and retained the ability to generate organoids in vitro and tumors in vivo, escaping the ADT-induced senescence observed in NCAM1- cells. By global transcriptional profiling RNASeq analysis of NCAM1-overexpressing LNCaP cells, fo
IDENTIFICATION OF NCAM1 AS A NOVEL PROGNOSTIC PROSTATE CANCER STEM CELL BIOMARKER / A. Castiglioni ; tutor: P. P. Di Fiore ; phd coordinator: S. Minucci. Dipartimento di Oncologia ed Emato-Oncologia, 2022 Dec 16. 34. ciclo, Anno Accademico 2022.
UNRAVELING TAZ REGULATION IN BREAST CANCER BY CRISPR SCREENING
YAP and TAZ are transcriptional coactivators belonging to the Hippo signaling pathway. Their activity is regulated by a complex network of upstream signals. As consequence, they orchestrate a wide spectrum of downstream cellular responses. In physiological conditions, a precisely balanced equilibrium governs activation and inhibition of YAP/TAZ. Disruption of this equilibrium unleashes YAP/TAZ oncogenic potential, promoting the development and progression of several human malignancies. In particular, TAZ overexpression has been associated with increasing aggressiveness of breast tumors, endowing mammary gland cells with oncogenic features, as stemness properties, migration, invasion and chemoresistance. Yet, YAP/TAZ mutations or genetic alterations of their known upstream regulators are relatively rare, raising the question of how YAP/TAZ are upregulated in cancer cells. To gain insights into novel TAZ regulators within the context of breast cancer, we conducted a series of loss-of-function CRISPR/Cas9 pooled genetic screens in vitro and in vivo on a selected subset of genes, previously identified in our laboratory as modulators of TAZ transcriptional activity. With this study we aimed to identify genes crucial for breast cancer cells growth, start to investigate their mechanism of action and explore their potential functional relationship with YAP/TAZ dysregulation in cancer. Among the identified hits, we focused our attention on FERMT2 and performed an initial characterization of its pro-tumorigenic role as TAZ activator.
UNRAVELING TAZ REGULATION IN BREAST CANCER BY CRISPR SCREENING / A. Chiesa ; internal advisor: P.G. Pelicci ; tutor: S. Campaner ; phd coordinator: S. Minucci. Dipartimento di Oncologia ed Emato-Oncologia, 2023 Dec 12. 35. ciclo, Anno Accademico 2023.
STATISTICAL METHODS TO ASSESS ROCKFALL SUSCEPTIBILITY IN AN ALPINE ENVIRONMENT: A FOCUS ON CLIMATIC FORCING AND GEOMECHANICAL VARIABLES
The overarching goal of the doctoral thesis was thus the development of a systematic procedure capable to examine and enhance the role of geomechanical and climatic processes in rockfall susceptibility, performed with statistically based and Machine Learning techniques. To achieve this purpose, two case studies were analysed in the Italian Alps (Valchiavenna, Lombardy Region; Mountain Communities of Mont Cervin and Mont Emilius, Aosta Valley Region).For both case studies, Generalized Additive Models (GAM) were used for rockfall susceptibility assessment; for the Valchiavenna case study, a Random Forest (RF) model was tested too. All models were validated through k-fold cross validation routines and their performance evaluated in terms of area under the receiver operating characteristic curve (AUROC). Predictors’ behaviour physical plausibility was verified through the analysis of the mathematical functions describing the predictors-susceptibility modelled relationships. Specific objectives of the two case studies differed.The Valchiavenna case study was dedicated to testing the role of the outcrop-scale geomechanical properties in a rockfall susceptibility model. Specific objectives were: (i) the optimal selection of sampling points for the execution of geomechanical surveys to be integrated within an already available dataset; (ii) the regionalization over the study area of three geomechanical properties, namely Joint Volumetric Count (Jv), rock-mass weathering index (Wi) and rock-mass equivalent permeability (Keq); (iii) the implementation of the regionalized properties as predictors in a rockfall susceptibility model, along with the traditional morphometric variables; (iv) the investigation of prediction limitations related to inventory incompleteness; (v) the implementation of a methodology for the interpretation of predictors’ behaviour in the RF model, usually considered a black box algorithm; (vi) the integration of the RF and GAM outputs to furnish a spatially distributed measure of uncertainty; (vii) the exploitation of satellite-derived ground deformation data to verify susceptibility outputs and interpret them in an environmental management perspective. The additional geomechanical sampling points were selected by means of the Spatial Simulated Annealing technique. Once collected the necessary geomechanical data, regionalization of the geomechanical target properties was carried out by comparing different deterministic, regressive and geostatistical techniques. The most suitable technique for each property was selected and geomechanical predictors were implemented in the susceptibility models. To verify rockfall inventory completeness related effects, the GAM model was performed both on rockfall data from the official landslide Italian inventory (IFFI) and on its updating with a field-mapped rockfall dataset. Regarding the RF model, the Shapely Additive exPlanations (SHAP) were employed for the interpretation of the predictors’ behaviour. A comparison between GAM and RF related outputs was carried out to verify their coherency, as well as a quantitative integration of the resulting susceptibility maps to reduce uncertainties. Finally, the rockfall susceptibility maps were coupled with Synthetic Aperture Radar (SAR) data from 2014 to 2021: a qualitative geomorphological verification of the outputs was performed, and composite maps were produced.The key results were: (i) geomechanical predictor maps were obtained applying an ordinary kriging for Jv and Wi (NRMSE equal to 13.7% and 14.5%, respectively) and by means of Thin Plate Splines for Keq (NRMSE= 18.5%). (ii) Jv was the most important geomechanical predictor both in the GAM (witha deviance explained of 7.5%) and in the RF model, with a rockfall susceptibility increase in correspondence of the most fractured rock masses. (iii) Wi and Keq were penalized (i.e., they had low influence on rockfall susceptibilit
STATISTICAL METHODS TO ASSESS ROCKFALL SUSCEPTIBILITY IN AN ALPINE ENVIRONMENT: A FOCUS ON CLIMATIC FORCING AND GEOMECHANICAL VARIABLES / G. Bajni ; tutor: T. Apuani ; co-tutor: C.A.S. Camera ; coordinatore: M.I. Spalla, F. Camara Artigas. Dipartimento di Scienze della Terra Ardito Desio, 2022 Mar 23. 34. ciclo, Anno Accademico 2021. [10.13130/bajni-greta_phd2022-03-23].
THE VALUE-BASED HEALTH CARE: A PSYCHOLOGICAL PERSPECTIVE IN CANCER CARE
This project has been developed in the breakthrough framework of the Value Based Health Care (VBHC), which aims at improving health outcomes at lower cost in different medical conditions. The VBHC paradigm has been applied in cancer care to improve the management of the disease, considering epidemiological, medical, psychological and economic outcomes. Two studies have been designed and implemented to better investigate the psychological perspective of lung and prostate cancer patients, in the light of the VBHC paradigm. The project identifies the trend of clinical and psychological status over time and predict this change with sociodemographic or medical variables. Several categories of patients characterized by different trends were identified, elucidating the psychological dimension of cancer patients. A person-oriented approach was used to analyze patients’ recovery. One-year Quality of Life (QoL) trends in early stage of lung and prostate cancer patients undergoing surgery were identified. Patients’ recovery after surgery was characterized by both an overall decrease of symptoms and an increase of health and functioning over time. Prostate cancer patients showed different longitudinal trajectories of urinary incontinence and sexual dysfunction. In lung cancer patients, pre-surgery QoL, type of surgery, perioperative complications, and age, affected the post-surgery QoL as well as the linear and quadratic trends over time. Through the implementation of this holistic approach, the predictive model of patients’ recovery will be developed, thus improving medical decision-making, the choice of treatment, and patients’ awareness about their care process. Potential harms, QoL, and expected outcomes will be more predictable and better manageable. Participants will be more empowered, being more aware of their post-surgical care.
THE VALUE-BASED HEALTH CARE: A PSYCHOLOGICAL PERSPECTIVE IN CANCER CARE / C. Marzorati ; supervisor: G. Pravettoni ; internal advisor: K. Mazzocco. DIPARTIMENTO DI ONCOLOGIA ED EMATO-ONCOLOGIA, 2020 Jan 28. 31. ciclo, Anno Accademico 2019. [10.13130/marzorati-chiara_phd2020-01-28].
INTO THE STRUCTURE OF THE PERMIAN DEEP CONTINENTAL CRUST: A MULTISCALE APPROACH TO RECONSTRUCT MIGMATIZATION IN THE VALPELLINE SERIES (DENT BLANCHE TECTONIC SYSTEM, WESTERN ITALIAN ALPS)
Granulite and migmatite are widespread across different tectonic settings, such as those associated with collisional crustal thickening during orogenic events and post-collisional or back-arc lithospheric thinning. Studying these rocks provides an exceptional opportunity to observe the deep portions of the continental lithosphere and infer insights into the processes that control its evolution. In extensional settings, orogenic root delamination and lithospheric thinning cause asthenospheric upwelling, increasing heat flow and enhancing partial melting of the lithospheric mantle and deep crust. During the Permian, the Variscan lithosphere was affected by extension due to the Pangea break up followed by Tethys opening. This Thesis investigates the evolution of the Valpelline Series in the Dent-Blanche Tectonic System (Western Italian Alps) which represents an exceptionally preserved exposure of Permian deep continental crust, despite being involved in the subsequent Alpine-age reworking. It shows migmatitic metapelite with different mineral assemblages (i.e., garnet-, cordierite- and orthopyroxene-bearing), amphibolite and marble, intruded by aplite and pegmatite. Melt production in metapelite and locally in amphibolite occurred during the development of the regional foliation. Quantitative microstructural and mineral-chemical analyses allow identifying three tectono-metamorphic stages. The first (D1) is associated with an early foliation (S1) preserved within mafic boudins enclosed in migmatitic gneiss. The second (D2) is related to the dominant foliation in migmatitic gneiss (S2), coeval with the regional scale anatexis and growth of garnet, orthopyroxene and cordierite. The third (D3) is related to the late folding of S2 and the development of a sillimanite-rich axial plane foliation (S3) which wraps around garnet and cordierite. Meso- and microstructural analysis helped to identify melt migration pathways in the deep continental crust. These pathways, corresponding to high-strain zones such as the S3 sillimanite-rich foliation, exhibit distinct mineralogical and structural features indicative of melt-rock interaction processes during deformation. P–T conditions of migmatization in the Valpelline Series, recorded in different lithologies, are contrasting. Black amphibolite and orthopyroxene-migmatitic gneiss show peak conditions comprised between 800–900 °C and 0.8–1.0 GPa, while cordierite-migmatitic gneiss registered peak P–T conditions at 750–800 °C and ~0.5 GPa. These contrasting P–T conditions are associated with different ages. Multi-method geochronology revealed that migmatization peaked during the D2 stage at ~290 Ma and continued until ~285–280 Ma over a period of at least 5 Ma, as suggested by garnet diffusion modelling. The reconstructed tectono-metamorphic history reflects an evolution that originated from Variscan post-orogenic collapse and lithospheric extension during the late Carboniferous-early Permian.
INTO THE STRUCTURE OF THE PERMIAN DEEP CONTINENTAL CRUST: A MULTISCALE APPROACH TO RECONSTRUCT MIGMATIZATION IN THE VALPELLINE SERIES (DENT BLANCHE TECTONIC SYSTEM, WESTERN ITALIAN ALPS) / F. Caso ; tutor: M. Zucali ; co-tutor: F. Farina ; coordinatore: M.I. Spalla. Dipartimento di Scienze della Terra Ardito Desio, 2024 Jan 27. 37. ciclo, Anno Accademico 2023/2024.
HYDROXYAPATITE-BASED MATERIALS FOR ENVIRONMENTAL PROCESSES
In this dissertation, hydroxyapatite-based and hydroxyapatite with carbon composite materials have been rationally designed, synthesized, characterized and applied to processes of environmental concern, namely wastewater remediation (as sorbents) and electrochemical CO2 reduction reaction, CO2RR (as electrocatalysts).Pristine hydroxyapatite (HAP, Ca10(PO4)6(OH)2) and composite HAP/carbon materials have been synthesized according to a simple wet co-precipitation route. Composition, structure and morphology of samples have been investigated by a plethora of physical-chemical techniques (i.e. ICP/OES elemental analyses, N2 adsorption/desorption isotherms, XRD, TEM (and STEM) imaging, TEM/EDX mapping, transmittance FT-IR and Raman spectroscopies, and XPS). Since adsorption onto materials’ surface is crucial in both sorption and catalytic applications, a special attention has been devoted to surface properties, determined by means of zeta potential measurements, gas-solid calorimetric/volumetric titrations, and liquid-solid volumetric titrations, thus assessing the amphoteric nature of HAP-based surfaces.At first, pristine pristine HAP has been studied as sorbent for the remediation of inorganic pollution in wastewater (i.e. heavy metal cations such as Cu(II), Pb(II), Cr(III), Ni(II) and Co(II)). Stirred batch adsorption tests and collection of adsorption isotherms in solutions containing both individual and mixtures of heavy metal cations allowed to quantitatively assess HAP sorption ability. Ad-hoc experiments (i.e. microcalorimetric adsorption isotherms) and physical-chemical investigation of metal-loaded samples shed light on the adsorption mechanisms of several polluting species onto HAP surface. Eco-friendly HAP/carbon composites from renewable resources have been instead applied to the remediation of more complex effluents, where simultaneous organic and inorganic pollution may occur. Thanks to their dual nature, HAP/carbon composites exerted outstanding adsorption performances towards both class of polluting species. Both in the form of pristine HAP and HAP/carbon composites, the sorbents exhibited a strict retention of adsorbed pollutants when undergoing leach testing, therefore ensuring no secondary pollution issues.In an exploratory study, HAP/carbon composites have been also implemented as modifiers in self-standing and mechanically stable electrodes for the electrochemical detection of heavy metal cations traces in waterbodies. Benefitting from the pronounced HAP affinity towards such species, encouraging results have been obtained on the detection of some benchmark pollutants, although the registered limits of detection (ca. one order of magnitude improvement to match legislative requirements) and linearity ranges currently limit HAP/carbon modified electrodes applications.Finally, during a 6-months stay at University of California Irvine (UCI), hosted by Prof. Plamen Atanassov group, HAP has been applied as dopant in CO2RR electrocatalysts. The peculiar ability of HAP amphoteric surface to adsorb and destabilize CO2 (i.e. breakage of its linearity) has been exploited in composite electrocatalysts, composed by copper nanoparticles (active phase) and HAP (dopant), supported on N-doped 3D assembly of graphene nanosheets (3D-GNS, carbonaceous support). When tested in a lab scale CO2 electrolyzer, HAP-doped electrocatalysts displayed a faradic efficiency (FE) towards CO2RR ≥ 80% throughout the whole potential range under investigation, minimizing parasitic Hydrogen Evolution Reaction (HER). Interestingly, HAP doping actually altered CO2RR product distribution: in particular, small quantities of C2+ products were produced under low applied overpotential while at higher voltage formate production was boosted. Product distribution alteration has been rationalized taking into account the ability of HAP acid-basic surface groups to stabilize CO2 reduction intermediate
HYDROXYAPATITE-BASED MATERIALS FOR ENVIRONMENTAL PROCESSES / M. Ferri ; tutor: Prof. A. Gervasini ; co-tutor: S. Trasatti, P. Cristiani ; coordinator: D. Roberto. Dipartimento di Chimica, 2021 Mar 22. 33. ciclo, Anno Accademico 2020. [10.13130/ferri-michele_phd2021-03-22].
ROLE OF CIRCULAR PVT1 IN ISCHEMIC HEART FAILURE
Circular RNAs (circRNAs) are involved in the pathogenesis of several cardiovascular diseases (CVDs), including heart failure (HF). circRNAs can bind microRNAs (miRNAs), acting as competing endogenous RNAs (ceRNA), thus regulating the expression levels of miRNA target mRNAs. However, these interactions and their respective functions in ischemic HF (IHF) remain largely unknown. This study aimed to identify circRNA-based ceRNA networks in non-end-stage IHF. Transcriptome analysis of left ventricles of IHF patients identified significant differentially expressed circRNAs, miRNAs and mRNAs. Potential miRNAs interacting with validated circRNAs as well as mRNA targets of sponged miRNAs were determined using bioinformatics tools. Predicted miRNAs and mRNAs were filtered for their dysregulation in IHF, resulting in 662 circRNA-miRNA-mRNA interactions. KEGG terms of the identified interacting mRNAs indicated CVD-related signaling pathways. circPVT1 emerged as one of the main hubs of circRNA-miRNA-mRNA network in IHF. RNA-Seq analysis following circPVT1 knockdown in immortalized human cardiomyocytes identified differentially expressed genes mainly involved in fibrosis and in cellular senescence. Interestingly, circPVT1 expression significantly increased in cardiac fibroblasts after TGF-β1 treatment. circPVT1 silencing attenuated the levels of pro-fibrotic markers induced by TGF-β1, highlighting a pro-fibrotic role of circPVT1. RNA pull-down assays confirmed the interaction between circPVT1 and three fibrosis-related miRNAs, miR-30a-5p, miR-125b-5p and miR-369-5p. The levels of these miRNAs were not altered upon circPVT1 knockdown. However, the expression of their mRNA targets was deregulated upon circPVT1 silencing, suggesting that circPVT1 modulate miRNA cellular bioavailability. Inhibition of either miR-30a-5p or miR-125b-5p, but not miR-369-5p, significantly reversed the reduced expression of TGF-β1-induced pro-fibrotic markers following circPVT1 silencing, indicating that both miR-30a-5p and miR-125b-5p act as downstream effectors of circPVT1 in cardiac fibroblast activation. Our findings suggest that circPVT1 has a pro-fibrotic function and regulate cardiac fibroblast activation via miR-30a-5p and miR-125b-5p sponging.
ROLE OF CIRCULAR PVT1 IN ISCHEMIC HEART FAILURE / A. Bibi ; tutor: A. Barbuti ; co-tutor: F. Martelli ; phd school coordinator: R. Mantovani. Dipartimento di Bioscienze, 2024 May 07. 36. ciclo
PARATHYROID HORMONE ACTIVITY AND RELATED CLINICOPATHOLOGICAL ALTERATIONS IN DOGS AFFECTED WITH CHRONIC KIDNEY DISEASE
Renal secondary hyperparathyroidism (RHPT) is a common complication in humans and dogs affected with chronic kidney disease (CKD). RHPT is promoted mainly by the loss of nephrons and consequent hyperphosphatemia and its severity increases in parallel to the disease. The identification and management of RHPT are suggested in dogs affected with CKD, to establish promptly appropriate treatments and follow the progression of the disease. However, in clinical practice, the determination of parathyroid hormone (PTH) concentration is not routinely due to limited available and validated method and need of radioimmunoassay. In CKD, data about the presence of RHPT in large populations of dogs and the thorough investigation of the association of PTH and other clinicopathological parameters involved in kidney disease are limited. Moreover, in human medicine, PTH has also been identified as an independent prognostic factor in patients affected with CKD and it is also reported that dysregulation of mineral metabolism leading to RHPT is present also in course of acute kidney injury (AKI), as occurs in CKD. Information about these two aspects in dogs is lacking.Hence, this thesis has been divided into two main parts: the first about analytical aspects and the second about clinical data.The first part was focused on the analytical validation of a new immunoenzymatic method validated in humans for the measurement of canine intact PTH and the establishment of the reference interval in healthy dogs.The second main part of this thesis was aimed to investigate the prevalence of RHPT in dogs affected by CKD and AKI, to compare these populations, and to assess the association between PTH and hematological, biochemical, and urinary parameters. Finally, a survival study has been done to elucidate the possible role of PTH as a survival factor in dogs with CKD.The first part of the study has allowed us to successfully validate the method in dogs, which has showed satisfying accuracy and repeatability, and to demonstrate a storage stability of PTH until 6 months at freezing temperature. A reference interval of 1.42-14.59 pg/mL has been established.The second part has allowed us to confirm the relevant presence of RHPT in CKD, also in early stages, and discover several associations between PTH and some parameters such as creatinine and phosphorus, as known, and others not yet identified, such as potassium, hematocrit, and blood pH. RHPT has been identified also in a high percentage of dogs affected with AKI, though the PTH concentrations were not significantly different between AKI and CKD. The survival analysis has revealed that higher concentrations of PTH corresponded to significantly short survival times compared to the lower concentrations, showing a possible prognostic role of PTH in dogs.
PARATHYROID HORMONE ACTIVITY AND RELATED CLINICOPATHOLOGICAL ALTERATIONS IN DOGS AFFECTED WITH CHRONIC KIDNEY DISEASE / J. Zambarbieri ; tutor: P. Scarpa ; coordinatori: F. Ceciliani, V. Grieco. Università degli Studi di Milano, 2022 Sep 06. 34. ciclo, Anno Accademico 2021.
DIETARY BIOACTIVES IN THE MODULATION OF METABOLIC, FUNCTIONAL AND AGING BIOMARKERS
Bioactive compounds and bioactive-rich foods play a crucial role in modulating key physiological pathways related to human health, including cardiometabolic regulation, intestinal function, and healthy aging. In particular, dietary (poly)phenols and xanthines have gained attention for their potential benefits on several metabolic and functional biomarkers. However, the mechanisms are not completely understood and the evidence in humans are still scarce. The present Ph.D. thesis investigates the role of dietary bioactives and bioactives-rich foods in modulating functional biomarkers of human health, with a focus on the mechanisms involving lipid metabolism, oxidative stress and inflammation, intestinal permeability, and telomere integrity. The research was structured into three main chapters, combining both in vitro and in vivo approaches.In Chapter 1, the mechanisms of action of selected bioactives, including (poly)phenols and xanthine-derived metabolites, were explored in different cellular models. In particular, 3T3-L1 adipocytes and Caco-2 intestinal epithelial cells were used to investigate processes such as lipid accumulation, glucose metabolism, and intestinal barrier integrity. Additionally, in 3T3-L1 adipocytes, a systematic review highlighted the modulatory role of dietary (poly)phenols on adipocyte metabolism, showing that different classes of (poly)phenols can influence key markers involved in lipid and glucose metabolism (e.g., AMPK, SIRT1). Experimental studies further revealed that quercetin-derived metabolites can reduce glucose-stimulated lipid accumulation, while blueberry and raspberry extracts protect intestinal barrier function under pro-inflammatory conditions by preserving tight junction integrity and reducing oxidative stress. Additionally, xanthine metabolites demonstrated protective effects on age-associated intestinal barrier dysfunction, suggesting their potential contribution to gut health during aging.Chapter 2 translated these mechanistic insights into the clinical setting, evaluating the effects of blueberry consumption, as source of dietary bioactives such (poly)phenols, in a human dietary intervention study. The human trial (the BLUMET study) assessed the impact of an 8-week blueberry intervention on biomarkers related to cardiometabolic health, oxidative stress, inflammation and intestinal permeability in individuals with metabolic syndrome. Preliminary findings suggest that blueberry intake may modulate some biomarkers of adipose tissue metabolism such as adiponectin and slightly reduce total cholesterol, supporting its potential role in the protection of human health. Conversely, no significant findings were documented for the rest of the biomarkers analyzed suggesting the need for further investigations. In Chapter 3, the focus was placed on telomere biology, a well-established hallmark of aging. The work was carried out as part of an Erasmus+ traineeship program at the University of Copenhagen (Denmark). The experiments investigated the role of xanthine and (poly)phenol metabolites in human samples (PBMCs) from the BLUMET study, as well as in cell culture models (THP-1). All activities were conducted under the supervision of Professor Peter Møller at the Department of Public Health, Section of Environmental Health, University of Copenhagen. In particular, cellular experiments, consisting in a dysmetabolism-induced monocytic cell line, demonstrated the potential of (poly)phenol metabolites to prevent telomere shortening suggesting their ability to modulate key pathways involved in cellular senescence. In addition, the findings from the BLUMET study suggested a possible protective effect of wild blueberry consumption on telomere length in individuals with metabolic syndrome, linking nutritional interventions to the preservation of genomic stability.In conclusion, this thesis contributes to the growing body of evidence supporting the
DIETARY BIOACTIVES IN THE MODULATION OF METABOLIC, FUNCTIONAL AND AGING BIOMARKERS / M. Rendine ; tutor: C. Del Bo' ; co-tutor: D. Martini ; coordinator: F. Cheli. Dipartimento di Scienze per gli Alimenti, la Nutrizione e l'Ambiente, 2025. 38. ciclo, Anno Accademico 2024/2025.
IMPACT OF INDOOR AIR POLLUTION ON INFLAMMATORY BIOMARKERS AND CARDIAC AUTONOMIC CONTROL IN HEALTHY SUBJECTS AND PATIENTS WITH CONGESTIVE HEART FAILURE (APICE STUDY)
The present study explores the modifications of cardiac autonomic control (CAC) during sleep and of systemic inflammatory profile associated with exposure to indoor air pollution (IAP) in healthy subjects and patients affected by chronic heart failure (HF).Twenty healthy volunteers and nineteen HF patients were enrolled. Indoor levels of fine particulate matter (PM2.5), nitrogen dioxide (NO2) and volatile organic compounds (VOCs) were monitored using a portable detector for 7 days. Together, a 7-day monitoring was performed through a wireless patch that continuously recorded electrocardiogram, respiratory activity and actigraphy. Indexes of CAC during sleep were derived from the biosignals: heart rate and low-frequency to high-frequency ratio (LF/HF) index of sympathovagal balance with higher values corresponding to a predominance of the sympathetic branch. Cyclic variation of heart rate index (CVHRI events/hour) during sleep, a proxy for the evaluation of sleep apnoea, was assessed for each night. After the monitoring, blood samples were collected to assess the inflammatory profile. Regression and correlation analysis were performed.A positive association between CVHRI during sleep and indoor air pollutants was found, with VOCs affecting healthy subjects (Δ%=+0.2% for 1µg/m3 VOCs, p=0.008) and NO2 impacting patients (Δ =+0.86 events/h for 10% increase in NO2, p=0.022. The CVHRI was also positively associated with LF/HF during sleep, thus higher CVHRI values corresponded to a shift of the sympathovagal balance towards a sympathetic predominance in healthy subjects (r=0.52; p=0.018). Indoor PM2.5 exposure was positively correlated with elevated plasmatic levels of TNF-α in patients (Δ%=17.18 for 10% increase in PM2.5, p=0.006). Indoor NO2 exposure levels were associated with increased plasmatic pro-inflammatory (sTREM-1) and anti-inflammatory (IL-10) biomarkers in healthy subjects (Δ%=+1.2% and Δ%=+2.4%, for 1µg/m3 NO2; p=0.005 and p=0.022, respectively), alongside upregulated expression of pro-inflammatory (IL-6 Δ%=8.94% for 10% increase in NO2, p=0.026; TREM-1 Δ%=6.60% for 10% increase in NO2, p=0.025) and anti-inflammatory (IL-10 Δ%=4.18 for 10% increase in NO2, p=0.037) pathways in patients.The study highlights a possible causal relationship between IAP exposure and higher risk of sleep apnoea events, associated with impaired CAC during sleep, and an altered inflammatory state both in healthy subjects and patients. The findings of this study have significant implications for both clinical practice and public health strategies, particularly in the context of improving cardiovascular health by addressing indoor air quality. The IAP could emerge as a critical and often neglected risk factor for the public health that can be addressed through targeted preventive interventions.
IMPACT OF INDOOR AIR POLLUTION ON INFLAMMATORY BIOMARKERS AND CARDIAC AUTONOMIC CONTROL IN HEALTHY SUBJECTS AND PATIENTS WITH CONGESTIVE HEART FAILURE (APICE STUDY) / A. Carandina ; tutor: E. Tobaldini, M. Bonzini; coordinatore: C. La Vecchia, F. Parazzini. Dipartimento di Scienze Cliniche e di Comunità, 2025. 37. ciclo, Anno Accademico 2023/2024.
INFERRING TUMOR PROPERTIES FROM GENE EXPRESSION DATA
The wide availability of transcriptomics data, along with the advancement of experimental technologies, computational tools, and statistical methods like machine learning (ML), has significantly improved the use of transcriptome data in cancer research, benefiting both diagnostic and prognostic clinical applications.Gene expression is used to predict the tissue of origin (TOO) of tumor samples, which is relevant in several scenarios such as the diagnosis of cancer of unknown primary (CUP), early cancer detection by means of circulating tumor cells (CTCs) or cell-free RNA (cfRNA) and the identification of mislabelled cell lines. So far those problems have been addressed separately but we ask if one classifier is sufficient to solve them all and investigate what are the determinants of cancer origin prediction performance. To this end, we develop TOPOS (Tissue of Origin Predictor of Onco-Samples), a ML classifier that can distinguish 15 different tissues and we test it on more than 36,000 samples spanning primary tumors, metastases, CTCs, patient-derived xenografts (PDXs), cancer cell lines and organoids. We demonstrate that by adopting a specific training composition and data processing approach, we can achieve high performance across different onco-types, including CTCs, suggesting the applicability of gene expression data in liquid biopsy.From cancer transcriptomic profiles we can extract features that hold information about tumors’ prognosis. To identify prognostic biomarkers, statistical methods like Cox regression have been adapted from survival analysis techniques. However, these methods rely on assumptions that may not always hold and restrict charting of feature-survival interaction, like for instance non-monotonic relationships, which have been shown to exist for mutation load and chromosomal instability (CIN). We wonder whether a similarly non-monotonic relationship occurs also at gene expression level and demonstrate that for some genes a poor or favorable outcome is found at a medium transcription level and that the shape of gene expression distribution plays a role in determining which expression range is more likely to be associated with good or bad patient prognosis.In this study, we have examined a diagnostic and prognostic application of gene expression data and explored how computational techniques can enhance its use in cancer research.
INFERRING TUMOR PROPERTIES FROM GENE EXPRESSION DATA / D. Cagnina ; tutor: M. Schaefer co-tutor: S. Santaguida. Dipartimento di Oncologia ed Emato-Oncologia, 2025 Jan 21. 36. ciclo, Anno Accademico 2023/2024.
CLANKRIMINALITÄT: LE RAPPRESENTAZIONI SOCIALI E ISTITUZIONALI DI UN FENOMENO CRIMINALE IN EMERSIONE NEL CONTESTO BERLINESE
"This research project entitled "" Clankriminalität in Berlin: the social and institutional representation of an emerging criminal phenomenon” aims at investigating a recent criminal phenomenon whose presence is found within an environment often considered unrelated to the emergence of forms of organized crime: the German context. The theoretical framework of the research is structured around two thematic areas that are strongly interconnected with each other, namely the processes and dynamics of emergence of complex forms of organized crime on the one hand, and the controversial relationship existing between migratory processes and organized crime on the other hand. The study is carried out taking into consideration different sources that provide a representation of the phenomena, such as BKA and LKA reports, the archives of German newspaper and the scientific literature pubblished on the topic. The aforementioned sources are integrated with interviews to experts and privileged observers who cover a variety of roles, in order to ensure the heterogeneity of the sample and obtain multiple and diversified information. The ultimate goal is to put forward hypotheses about the factors that contributed to the construction of the social and institutional representation of the phenomenon."
CLANKRIMINALITÄT: LE RAPPRESENTAZIONI SOCIALI E ISTITUZIONALI DI UN FENOMENO CRIMINALE IN EMERSIONE NEL CONTESTO BERLINESE / S. Mazzenzana ; tutor: M. Massari ; coordinatore: F. Basile. Dipartimento di Studi Internazionali, Giuridici e Storico - Politici, 2022 Jul 11. 34. ciclo, Anno Accademico 2021.
TACKLING BIOFILM-RELATED OPPORTUNISTIC FUNGAL INFECTIONS
More than 1.5 million people/year die for a biofilm-related fungalinfection. The ability to develop biofilms is widespread amonghuman opportunistic pathogens, among them fungi. Biofilms are acommunity of microorganisms with the ability to attach to surfacesbiotic or abiotic, surrounded by a self-produced extracellularpolymeric matrix. The pathogen clearance and the generalovercoming of the infection are complicated by biofilm presence asit confers tolerance to antimicrobial treatments. Compromised oraltered immune response and an inflammatory milieu canexacerbate the pathogenesis and prolong the resolution of theinfection. Candida albicans, between yeasts, and Aspergillusfumigatus between molds, are pathobionts considered the mostcommon agents of opportunistic fungi mediated. In our study weassessed different strategies to eradicate biofilms and counteractbiofilm-related infections, targeting both pathogens and hostimmune response. We investigated the efficacy of urinary fractionsafter cranberry extract intake by healthy volunteers against C.albicans biofilm. By mass spectrometry analyses, we identified twometabolites picking in the most active urine fractions, 5-(3′,4′-dihydroxy phenyl)-γ-valerolactone and 4-hydroxybenzoic acid,which revealed a strong inhibitory effect on C. albicans adhesionand biofilm formation. Both compounds were also able todownregulate the expression of key genes involved in early phasesof biofilm formation. Due to the spread of the use of catheters orprostheses in the medical field, the possibility to developbiomaterials with biofilm inhibitory activity could represent a good10strategy to control nosocomial infections. We thus investigatednewly synthesized hydrogels and demonstrated their ability toprevent C. albicans growth.To target both pathogen virulence and host immune response, weconducted two different studies, the first on C. albicans systemiccandidiasis and the second on A. fumigatus infection in cysticfibrosis patients.We investigated the possible dual action of pilocarpine, amuscarinic receptor agonist, in vitro and in vivo using the modelhost Galleria mellonella. Pilocarpine showed a direct effect ininhibiting C. albicans biofilm biomass and metabolic activity, and itsadministration to infected larvae increased larval survival. Becauseof the different behavior of pilocarpine compared to acetylcholine inthe modulation of larva immune response, we concluded that theantifungal activity of pilocarpine might rely on muscarinic-likereceptor activation on C. albicans, whereas the strongimmunomodulatory effect of acetylcholine (but not of pilocarpine)might imply the engagement of nicotinic receptors. A similarapproach, aimed at targeting both infection and inflammation, wasused to investigate A. fumigatus persistence in CF patient lungs.Indeed, CF monocytes are characterized by altered intracellularlipid accumulation, that compromise pathogen clearance, and theinfection resolution. We demonstrated that the treatment of patientmonocytes with Myriocin, by inhibiting the synthesis of sphingolipidsand hampering CF inflammation, ameliorate the A. fumigatusconidia internalization and clearance. On the host side, Myriocin11was able to reduce the over expression of genes encoding for proinflammatorycytokines and at the same time to increase theexpression of gene encoding for pathogen recognizing receptors,crucial for its clearance.Our results indicate that, despite we are still far from clinicalpractice, exploring alternative anti-biofilm strategies could pave theway to new target discovery and to integrated approaches able topromote the infection resolution.
TACKLING BIOFILM-RELATED OPPORTUNISTIC FUNGAL INFECTIONS / E. Ottaviano ; tutor: E. Borghi ; coordinatore: M. Samaja. Dipartimento di Scienze della Salute, 2021 Jan 21. 33. ciclo, Anno Accademico 2020. [10.13130/ottaviano-emerenziana_phd2021-01-21].
GUANYLATE CYCLASE ACTIVATING PROTEIN 1 MONOMER-DIMER EQUILIBRIUM CONTROLLED BY CA2+ OR MG2+ BINDING: HINTS TO UNDERSTAND RETINAL GUANYLATE CYCLASE REGULATION
Neuronal calcium sensors play a crucial role in different pathways of Ca2+-mediated neurotransmission. Among them guanylate cyclase-activating protein 1 (GCAP1) is expressed only in photoreceptors and activates or inhibits retinal guanylate cyclase 1 (retGC1) depending on cellular Ca2+ concentrations during phototransduction. To date, 22 pathogenic mutations responsible for retinal dystrophy have been associated to GCAP1, but a complete picture of the molecular determinants of the disease is still missing. The only crystal structure available so far is the wt Ca2+-bound monomeric homologue from chicken and no cure exists for retinal dystrophy. In this work I report for the first time that the recombinant human GCAP1 is characterized by a highly dynamic monomer-dimer equilibrium, whose dissociation constant is influenced by salt concentration and by the nature of the divalent ion bound. Surprisingly, I discovered that also the chicken protein shows a similar mechanism, suggesting that this property could be potentially functional for GCAP1 activity and conserved among different species. Despite the large number of crystallization trials, no diffracting crystal of the human GCAP1 was obtained, probably due to the flexible C-terminal tail and the intrinsic dynamicity of the protein. To overcome this issue, I produced a construct lacking the 12 C-term residues and stabilized by a disulfide bridge between the N- and C-term domains which was successfully crystallized. We showed that such engineered construct is able to regulate retGC1 as well as the wt protein. By combining SAXS, protein-protein docking and molecular dynamics simulation we propose two novel three-dimensional models of Ca2+-bound GCAP1 dimer which are stabilized by some of the residues involved in the interaction with the retGC1. We used a biophysical and biochemical approach to thoroughly investigate three pathogenic variants (D100G, E155A and E155G) characterized by mutations in residues directly involved in Ca2+-coordination. All the three variants were able to form oligomers in solutions, showing a decreased affinity for Ca2+ and constitutively activating retGC1 at physiological calcium concentrations. Besides local structural effects, the mutations perturb also the oligomeric state of GCAP1 suggesting that the multimeric assembly of the protein could affect its proper biological function. A recombinant baculovirus for the expression of the cytoplasmic domain of retGC1 in insect cells was produced with the aim to get atomic structural information on the GCAP1/retGC1 complex. This will facilitate the identification of drug candidates able to recognize the binding region of the pathogenic GCAP1 mutants with the cyclase and to competitively inhibit the constitutive retGC1 activation, restoring the homeostasis of second messengers which is impaired in retinal degenerative diseases. A preliminary molecular docking based on the crystal structure of the chicken protein was performed and I identified four molecules able to bind the wt human GCAP1 in the millimolar/micromolar range.Together these results shed new light on the quaternary assembly of the wt human GCAP1, showing how the structural changes related to the presence of Ca2+ or Mg2+ are reflected in the different measured dimerization constant. Such conformational changes are in turn likely related to the regulatory mechanism of GCAP1 in the modulation of the retGC1 activity. The differences between the oligomerization state of D100G, E155G and E155A variants suggest a correlation between the altered quaternary assembly of GCAP1 and the aberrant activity of the mutants, representing a step forward to dissect the structural bases of the altered regulatory mechanism of GCAP1 in retinal dystrophies.
GUANYLATE CYCLASE ACTIVATING PROTEIN 1 MONOMER-DIMER EQUILIBRIUM CONTROLLED BY CA2+ OR MG2+ BINDING: HINTS TO UNDERSTAND RETINAL GUANYLATE CYCLASE REGULATION / F. Boni' ; tutor: M. Milani ; co-tutor: M. Bolognesi. Dipartimento di Bioscienze, 2021 May 20. 33. ciclo, Anno Accademico 2020. [10.13130/boni-francesco_phd2021-05-20].
ROLE OF ALOG FAMILY GENES IN INFLORESCENCE PATTERNING IN ORYZA SATIVA AND ARABIDOPSIS THALIANA
Inflorescence architecture is a key agronomical trait that determines fruit and seed yield. Understanding the genetic basis of inflorescence architecture will not only contribute to elucidate crop evolution/domestication mechanisms but also improve crop grain yield.Flowering plants develop different types of inflorescences, such as racemes in Arabidopsis and panicles in rice. The architecture is established during the early stages of reproductive development and it is determined by the activity of different meristem types and by the timing of the transition between indeterminate meristems to determinate ones. Inflorescence development is finely regulated by a genetic network that includes meristem identity genes and genes that regulate their expression; many genes are already known but others have still to be characterized to provide insight into how this complex process is controlled.Transcriptomic analysis performed in rice and in Arabidopsis through laser microdissection of different meristematic tissues highlighted differentially expressed genes belonging to the ALOG family suggesting their role in inflorescence patterning. We focus on G1L1, G1L2, and G1L5 of rice and on LSH1, LSH3, and LSH4 of Arabidopsis. G1L5 is already known to be a major regulator of inflorescence architecture, whereas LSH3 and LSH4 seem to have a role in meristem maintenance and organogenesis. Their expression profiles were analyzed by qRT-PCR and RNA in situ hybridization experiments using meristematic tissues from both species. We are also generating single and double/triple K.O mutants in different combinations by CRISPR-Cas9 genome editing technology to have a better understanding of their role in inflorescence patterning.The data so far obtained demonstrate the role of G1L1 and G1L2 in inflorescence branching and spikelet number determination and we also propose a role for G1L2 in root development. Furthermore, LSH1 seems to be involved in meristem maintenance and organ differentiation, and LSH3 in stem elongation. We propose the hypothesis that LSH1, LSH3, and LSH4 play a redundant function in inflorescence development.
ROLE OF ALOG FAMILY GENES IN INFLORESCENCE PATTERNING IN ORYZA SATIVA AND ARABIDOPSIS THALIANA / E. Franchini ; supervisore: M. Kater; coordinatore : M. Kater. Dipartimento di Bioscienze, 2020 Mar 05. 32. ciclo, Anno Accademico 2019. [10.13130/franchini-emanuela_phd2020-03-05].
PRINCIPIO DELLA DOMANDA E ASSORBIMENTO DEI MOTIVI NEL PROCESSO AMMINISTRATIVO DI LEGITTIMITA'
The research studies whether administrative courts must follow the order of the claims brought forward in administrative complaint (ricorso), as well as the related issue regarding to the possible integration of a claim into another.After a description of the long standing case law disagreement on such matters, resulted in the ruling of the Plenary Chamber (Adunanza plenaria) no. 5/2015, the study will highlight with adequate justification the insufficiency of the preator approach (and, even if in a minor extent, of the scholars’ theories) with respect to, in particular, the case in which the claimant has not brought its claims in a specific order.Without prejudice to the subjective nature of administrative rulings, case law and scholars, although they recognize (as it seems correct) binding attitude to conditional aggregations of grounds, deny any importance to the principle of correspondence between complaints and rulings (principio della domanda) with respect to the ordo procedendi whenever there is not a specific order of complaints brought forward.This research aims at demonstrating the – also in the last instance – in proceedings governed by the “principle of party disposition”, even if it is not acceptable that the first complaint assessed by the court is the one characterized by the most “radical” unlawfulness (solution proposed by the plenary chamber of the Council of State (Consiglio di Stato)), it is also not acceptable to have a prior assessment of the unlawfulness which would result in the most favorable outcome of the claimant (solution preferred in the literature). The only solution that seems to be consistent with the underlying premise is that courts must strictly follow, in the assessment of the claims, the order in which they have been articulated in the complaint.
PRINCIPIO DELLA DOMANDA E ASSORBIMENTO DEI MOTIVI NEL PROCESSO AMMINISTRATIVO DI LEGITTIMITA' / G. Lattanzi ; tutor: L.S. Bertonazzi ; coordinatore: D. U. Galetta. Università degli Studi di Milano, 2020 Nov 20. 32. ciclo, Anno Accademico 2019. [10.13130/lattanzi-giacomo_phd2020-11-20].
"A ""GREEN PATH"" STRATEGY FOR COMPLETE RECOVERY OF BUCKWHEAT PROCESSING BYPRODUCTS"
This PhD thesis fits in the general framework of the so-called “circular bioeconomy” and focuses on the valorization of buckwheat husk, an agro-industrial by-product generated during the milling process. The main objective of the work was to identify and develop innovative strategies for the reutilization of buckwheat husk, thereby contributing to the reduction of food waste and the establishment of more sustainable and resource-efficient production systems. This work aligns with one of the key priorities of the European Green Deal, namely the efficient and responsible utilization of food by-products and food waste.Throughout the three-year research period, the possible end uses of this by-product—containing interestring bioactives compounds, was investigated with the aim of promoting its sustainable reuse. The main research outcomes can be summarized as follows:1. A green extraction approach - based on microwave-assisted extraction (MAE)- was set up to improve recovery of the phenolics from buckwheat husk. The protocol developed here increased by 43.6% the polyphenols recovery compared to conventional techniques.2. Buckwheat husk was evaluated as a natural filler in biocomposite materials. By combining enzymatic action with a microwave-assisted treatment, an enhanced cellulose accessibility was achieved along with complete retention of the original antioxidant activity, making the resulting material of interest for bioactive packaging applications.3. Buckwheat husk was utilized as a substrate for solid-state fermentation (SSF) with the edible mushrooms Pleurotus ostreatus and Ganoderma annularis. Fungal growth improved the nutritional and functional profile of the husk, increasing its content in soluble dietary fiber and its proteolytic activity.4. Mixtures of husk and either mushroom biomass were successfully incorporated into bread formulations (at a 10% substitution level), without compromising technological performance. The SSF-derived ingredients show promise for the development of baked goods with a decreased glycemic index.Overall, this research demonstrates the multifunctional potential of buckwheat husk as a sustainable material and functional ingredient, contributing to the advancement of resilient, circular, and sustainable food systems.
"A ""GREEN PATH"" STRATEGY FOR COMPLETE RECOVERY OF BUCKWHEAT PROCESSING BYPRODUCTS / A.r. Speranza ; tutor: S. Iametti; cotutor: S. Limbo; coordinatore: D. Mora. - Milano. Dipartimento di Scienze per gli Alimenti, la Nutrizione e l'Ambiente, 2025. 38. ciclo, Anno Accademico 2024/2025."
STRATEGIC LITIGATION FOR WOMAN'S RIGHTS AT THE WORKPLACE: A LAW AND ECONOMICS PERSPECTIVE
Strategic litigation for woman‘s rights at the workplace: a Law and Economics perspectiveDespite comprehensive legal frameworks protecting gender equality in the workplace,significant disparities persist between men and women in pay, promotion opportunities, andcareer advancement across Europe. This thesis examines whether Strategic litigation caneffectively address these enduring inequalities.While existing scholarship focuses on formal legal protections and policy implementation,this thesis identifies a critical gap in understanding how informal institutions—cultural norms,stereotypes, and social expectations—perpetuate workplace discrimination despite robustlegal frameworks. The research addresses how Strategic litigation can target these deeperinstitutional barriers that traditional legal approaches fail to address.This thesis employs comparative legal analysis of Italy and Germany's gender equalityframeworks, combined with game theory and institutional analysis to examine the interactionbetween formal and informal institutions. This interdisciplinary approach analyzes landmarkStrategic litigation cases alongside contemporary workplace equality challenges. Theresearch demonstrates that Strategic litigation's effectiveness lies not merely in enforcingexisting legal rights, but in its capacity to reshape informal institutions and cultural normsthrough public engagement and media attention. The thesis reveals that while Europeanlegal frameworks provide adequate formal protections, strategic litigation can create newsocial equilibriums by challenging deeply embedded gender stereotypes and expectations.The study concludes that Strategic litigation offers a promising mechanism for achievingsubstantive gender equality by targeting both legal compliance and cultural transformation,particularly when supported by NGOs and backed by comprehensive public campaigns thatshift societal attitudes toward gender roles in the workplace.
STRATEGIC LITIGATION FOR WOMAN'S RIGHTS AT THE WORKPLACE: A LAW AND ECONOMICS PERSPECTIVE / J. Hartmann ; tutor: L. Violinico-tutor: V. Cecchini Manara ; coordinatrice del dottorato: D. U. Galetta. Dipartimento di Diritto Pubblico Italiano e Sovranazionale, 2025 Dec 11. 35. ciclo, Anno Accademico 2024/2025.
ONLINE LEARNING, UNIFORM CONVERGENCE, AND A THEORY OF INTERPRETABILITY
This doctoral thesis covers various aspects of theoretical machine learning relative to two of its most fundamental paradigms: batch learning and online learning. In particular, we address the role of feedback models for multiple online learning problems, the sample complexity for uniform convergence, and a learning-theoretic approach to interpretable machine learning. First, we focus on online learning and investigate variants of the multi-armed bandit problem, including settings with feedback graphs, expert advice, and delayed feedback. We improve bounds on the minimax regret for undirected, strongly observable feedback graphs and develop nearly optimal algorithms for directed, stochastic feedback graphs without prior information on the distribution of the graphs. Additionally, we derive improved regret bounds for bandits with expert advice and explore the impact of intermediate observations in the delayed feedback setting, designing a meta-algorithm to achieve near-optimal regret which shows a reduced effect of the total delay. Second, we study the uniform convergence property of real-valued function classes with finite fat-shattering dimension. We provide an improved bound on the sample complexity of uniform convergence, closing the gap with existing lower bounds. Finally, regarding interpretability, we establish a taxonomy for approximating complex binary concepts with interpretable models such as shallow decision trees. Leveraging uniform convergence for Vapnik-Chervonenkis classes and von Neumann's minimax theorem, we achieve a surprising trichotomy for interpretable concepts while revealing connections between interpretable approximations and boosting.
ONLINE LEARNING, UNIFORM CONVERGENCE, AND A THEORY OF INTERPRETABILITY / E. Esposito ; supervisor: N. Cesa-Bianchi ; co-supervisor: M. Pontil ; coordinatore: R. Sassi. Dipartimento di Informatica Giovanni Degli Antoni, 2024 Dec 05. 37. ciclo
DESIGNING FOUNDATIONAL STRATEGIES FOR DEVELOPING A FAMILY OF FOOD SYSTEMS SUSTAINABILITY ONTOLOGIES
During the last two decades, ontologies – theories for representing knowledge – have shown their potential in cohering heterogeneous perspectives and establishing common semantics across varied stakeholders and domains. Despite that, to date, limited and isolated efforts have been made with regard to sustainability – a participatory shared language would highly benefit in clarifying and possibly harmonizing the diverse perspectives, metrics, and approaches that currently exist on sustainability.To deal with those shortcomings, this PhD dissertation provides a toolkit of solid philosophical strategies establishing a suitable foundational basis to support the consistent and interoperable design of ontologies for sustainability. Food systems have served as the core case study of this work, due to being the primary drivers of many of the current global sustainability concerns threatening life on Earth, such as climate change, biodiversity loss, and malnutrition. The proposed toolkit covers three strategies: 1) a review analysis of currently available ontologies regarding sustainability and related theoretical challenges, a roadmap towards a family of interoperable sustainability ontologies, and a Sustainability Core Ontology (SCO), 2) a meta-framework for food systems sustainability, and 3) a methodology for developing food systems sustainability ontologies using the designed meta-framework, and a use-case ontology addressing sustainability in meat systems, named Sustainable Meat Systems Ontology (SuMSO). This PhD dissertation is a collection of three research papers, each dedicated to describing one of these strategies.
DESIGNING FOUNDATIONAL STRATEGIES FOR DEVELOPING A FAMILY OF FOOD SYSTEMS SUSTAINABILITY ONTOLOGIES / G.a. Ubbiali ; tutor: A. Borghinitutor: M. Langecoordinatore di Dipartimento: F. Guala. Dipartimento di Filosofia Piero Martinetti, 2025 Apr 29. 37. ciclo, Anno Accademico 2023/2024.
FEED & FOOD SAFETY: FROM ENVIRONMENTAL CHEMICAL CONTAMINANTS TO POTENTIAL EMERGING RISKS
The carry-over of chemical contaminants from feed to food products is a major concern in the animal production chain. For a proper containment, limits for feed as well as food products are fixed for a series of chemicals, e.g. dioxins and dioxin-like PCBs, heavy metals, some chlorinated pesticides, and some mycotoxins. Food and feed safety monitoring programs exist worldwide in order to protect consumers and animal health. Demand for animal products is expected to increase by up to 50% by 2050. Animal nutrition is the most challenging sector due to the limited availability of natural resources, climate change and food-feed-fuel competition. Due to the resource costs of conventional feeds, such as soybean meal, fishmeal, and standard grains such as maize, alternative sources (e.g. cereal processing by-products/co-products, former food products, seaweeds, insects) are increasingly being studied and used in animal diets. As new ingredients enter the food chain, new and potential risks are emerging on the feed and food security scene. At the same time, climate change can vary (and therefore increase) the risk for already widely known contaminants such as mycotoxins. An ideal goal is to assure that compliance to feed limits automatically results in compliance to food limits.The focus of this dissertation comprises two distinct parts: 1) in the first part (6 chapters), it focuses on the monitoring of different chemical contaminants (mycotoxins and antibiotics) in feed and food ingredients. Various subjects will be addressed concerning the multi-annual monitoring programs of several mycotoxins in northern Italy and the determination and monitoring of antibiotic residues in both feed and food, throughout different analytical techniques. Still on the subject of food safety, 2) the second part of this dissertation (3 chapters) deals with the issue of selenium supplementation in insects as an alternative means to compensate for selenium deficiency, taking into account, however, that Se represents a risk factor due to the narrow range between deficiency and toxicity.
FEED & FOOD SAFETY: FROM ENVIRONMENTAL CHEMICAL CONTAMINANTS TO POTENTIAL EMERGING RISKS / L. Ferrari ; tutor: L. Pinotti ; co-tutor: U. Vincent ; coordinator: F. Cheli. Dipartimento di Medicina Veterinaria e Scienze Animali, 2024 Mar 13. 36. ciclo, Anno Accademico 2023.
GABAA RECEPTOR AS A NOVEL REGULATOR OF PERIPHERAL PAIN SENSITIVITY AND LOCAL NEURON-GLIA INTERACTION
The pathogenesis of neuropathic pain and its chronicization process are still not fully elucidated. For this reason, therapies currently ongoing in the clinic are aimed to treat mostly the symptoms, considering poorly the target mechanisms causing those pathologies. The approach that we choose to investigate new pharmacological targets for the therapy of chronic pain, was to firstly investigate the mechanisms of nociception in physiological condition, finding new molecular pathways responsible for nociception modulation. Secondly, we were aimed to study the contribution of these targets in the pathological condition. In particular, in this PhD thesis, I focused on the characterization of the GABAergic system, primarily the GABAA receptors (GABAAR), along peripheral nociceptors axons (forming C-fibers), evaluating their putative role in the modulation of pain peripheral conduction, during different physio-pathological conditions. GABAAR mediate fast synaptic inhibition in the dendrites, soma and axons of mature neurons that maintain low intracellular Cl- concentrations. In immature neurons as well as mature somatosensory neurons, the intracellular Cl- concentration is physiologically elevated and this gives rise to depolarizing GABAAR currents. We here report that, in somatosensory unmyelinated C-fibre axons, GABAAR mediates depolarizing currents and that the magnitude and time course of GABAAR responses are determined by NKCC1 activity. GABA (1 µM-1 mM) depolarizing responses were mediated specifically by axonal GABAAR, indeed they were mimicked by muscimol (>1 µM) and gaboxadol (THIP; >1 µM) and blocked by bicuculline (50 µM). Depolarizing axonal responses to GABA were completely absent in mice lacking β3 GABAAR subunit (β3,fl/fl), in either all sensory neurons (AdvillinCRE) or in nociceptor neurons expressing NaV1.8 (snsCRE). qRT-PCR analyses suggested that the most common GABAAR composition in somatosensory DRG neurons was a2, b3 and g2. To examine the physiological role of axonal GABAAR, C-fibres were subjected to a sustained frequency challenge (3 min at 2.5 Hz) and three main effects were found. First, the amplitude of axonal GABA responses was increased, and this effect was deemed secondary to an NKCC1 mediated shift in E Cl-. Second, GABAAR activation increased the axonal conduction velocity of C-fibres. Third, axonal GABAAR was activated by endogenous ligands. Our results indicate that C-fibres sustained firing increases NKCC1 activity, shifting ECl- toward more positive values. In this condition, constitutive GABAAR currents maintain nociceptor conductance during sustained firing. Established the capability of peripheral GABAergic currents to stabilize nociceptor conductance during sustained activity, we investigated the GABAergic modulation in pathological conditions characterized by hypersensitivity of nociceptors. In this regard, we studied in vivo model of inflammatory pain on snsCRE;b3-/- mice. The outcomes indicated that GABAAR activity increment mechanical allodynia and prevent the insurgence of hyperalgesia. In addition, GABAAR activity prolonged the recovery time, maintaining the hypersensitive phenotype for a longer period of time (up to 4 weeks). This finding corroborated the hypothesis that GABAergic transmission within peripheral fibers is able to stabilize the physiological conduction of pain, although it appeared dangerous in pathological condition, promoting nociceptor hypersensitivity and chronicization. We found that ALLO, a neuroactive steroid able to activate GABAAR and endogenously synthesized in PNS, induces the release of the growth factor BDNF from SCs, which is able to target trkB receptors on axons, in turn inducing PKCe upregulation and activation. PKCe is a typical protein kinase known to be involved in the process of pain chronicization. Overall, these set of data suggested that GABAAR is involved in a complex paracrine mechanism med
GABAA RECEPTOR AS A NOVEL REGULATOR OF PERIPHERAL PAIN SENSITIVITY AND LOCAL NEURON-GLIA INTERACTION / V. Bonalume ; tutor: V. Magnaghi ; co-tutor: R. Carr ; coordinator: C. Sforza. DIPARTIMENTO DI SCIENZE FARMACOLOGICHE E BIOMOLECOLARI, 2020 Jan 20. 32. ciclo, Anno Accademico 2019. [10.13130/bonalume-veronica_phd2020-01-20].
THE POLITICS OF ECO-SOCIAL POLICIES: TOWARDS A JUST TRANSITION IN SPAIN AND IRELAND
Social and environmental challenges are deeply interconnected: it is indeed becoming increasingly evident how both climate change and green transition policies are giving rise to a new wave of social risk for the welfare state to address. Therefore, the so-called eco-social policies could represent suitable tools for the state to simultaneously pursue social and environmental goals in a synergic manner. This study concentrates on a particular type of eco-social policies, which are the ones designed to address the social consequences of decarbonisation and to achieve a just transition. Despite the importance that just transition policies could have in accompanying European countries’ net-zero transitions, these policies are not only still rarely diffused in reality but also undertheorized by political scientists, since the literature is currently dominated by prescriptive studies. Against this background, this Ph.D. thesis aims to contribute to the emerging debates about sustainable welfare and just transition by bringing in a so-far underdeveloped empirical-political perspective. Navigating a largely unexplored field, the thesis thus aspires to strengthen our analytical, theoretical and empirical understanding of just transition policies and related political dynamics. It does so, preliminarily, by proposing a novel conceptual-analytical framework; mapping existing just transition policies in European Union countries; and reflecting on the applicability of mainstream welfare state theories to potentially explain the emergence of just transition policies. Building on these preliminary insights, Spain and Ireland are selected as case studies for a comparison that aims to unveil the process culminating in the adoption and formulation of just transition policies in these two countries. The two cases are investigated through process tracing and qualitative methods, focusing on policy and politics analysis. In short, the thesis argues that national just transition policies are still relatively rare across the European Union and, with the notable exception of Spain, they are mostly formulated as narrow strategies, connected to investment-oriented instruments and to a governance that concentrates power in the hands of the government. The emergence of such policies can be explained as a result of a green-labour convergence among relevant socio-political actors engaging in political exchanges, with which political support for decarbonisation is traded for economic support to vulnerable societal groups.
THE POLITICS OF ECO-SOCIAL POLICIES: TOWARDS A JUST TRANSITION IN SPAIN AND IRELAND / M. Mandelli ; relatore: M.R.C. Jessoula ; coordinatore: M.R.C. Jessoula. Dipartimento di Scienze Sociali e Politiche, 2023 Sep 14. 35. ciclo, Anno Accademico 2022.
MICROALGAE PEPTIDES IN CARDIOVASCULAR DISEASE PREVENTION: STRUCTURE ELUCIDATION,BIOACTIVITY INVESTIGATION,AND IN SILICO MOLECULAR MODELING ANALYSIS
Bioactive food-derived peptides have been increasingly studied and the multiple health benefits they provide have been acknowledged. Since the most investigated sources of bioactive peptides are eggs, meat, fish, soybean, wheat, milk, and their derivatives or byproducts. More recently, some attention is being paid to microalgae, a promising unconventional protein source. On this basis, the PhD thesis was focused on the evaluation of the potential of microalgae to generate peptides with cardiovascular-promoting effects, especially hypotensive and antidiabetic activities, by targeting two therapeutic agents – angiotensin I converting enzyme (ACE) and peptidyl-peptidase IV (DPP-IV), respectively. To achieve this objective, multidisciplinary approaches were employed, involving peptidomic techniques to profile the peptide sequences, biochemical tools to analyze the bioactivity, and emerging molecular modelling methods to predict potentially bioactive peptides as well as to explore their possible mechanism of action. Briefly, the results showed that the protein hydrolysates from spirulina, PBP and chlorella generally presented significant ACE and/or DPP-IV inhibitory activities. By comparison, peptic hydrolysate of spirulina protein showed the best in vitro inhibiting effect on ACE with IC50 value of 0.1 ± 0.04 mg/mL while the tryptic hydrolysate of PBP stands out with the lowest IC50 value of DPP-IV inhibition (0.5 – 1.0 mg/mL). Noticeably, when working on the intestinal Caco-2 cells, all the hydrolysates turned to be less bioactive than in vitro, indicating their susceptibility to metabolic degradation by intestinal cells. This is further in line with the kinetics of DPP-IV inhibition of PBP tryptic hydrolysate working on the intestinal cells, which showed the decreasing trend of its bioactivity after incubation with Caco-2 cells for 3 h. This reflects the issue of peptide bioavailability, which could be stressed in further studies. Moreover, peptides Pep2 (FLKPLGSGK), Pep7 (QIYTMGK), Pep8 (FLFVAEAIYK), and Pep10 (QHAGTKAK) were screened from hydrolysates of chlorella protein by peptidomics combined with docking and MD. The modelling results indicated that they may block the important domain of both ACE and DPP-IV and generate dynamically stable peptide-ACE/DPP-IV complexes. Based on this theoretical evidence, further study will focus on the verification of their actual bioactivity by biochemical approaches.In conclusion, the bioactivity investigation of microalgae protein hydrolysates provides new evidence that microalgae protein are great sources to produce peptides with health-promoting properties. The exclusive data of peptide characterization makes a foundation to isolate single bioactive peptides as well as offers useful structural and functional implications for food ingredient formulation or pharmacological use.
MICROALGAE PEPTIDES IN CARDIOVASCULAR DISEASE PREVENTION: STRUCTURE ELUCIDATION,BIOACTIVITY INVESTIGATION,AND IN SILICO MOLECULAR MODELING ANALYSIS / Y. Li ; Faculty advisor: A. Arnoldi ; PhD coordinator: G. Aldini. Dipartimento di Scienze Farmaceutiche, 2021 Jun 14. 33. ciclo, Anno Accademico 2020. [10.13130/li-yuchen_phd2021-06-14].
L'ORDINE EUROPEO D'INDAGINE PENALE: DISCIPLINA NORMATIVA E PRIME ESPERIENZE APPLICATIVE
The European criminal investigation order, introduced by Directive 2014/41/EU, isan instrument of judicial cooperation in the field of evidence, which has becomenecessary, given the growing transnational dimension of crime as a result of thesublimation of geographical boundaries in the European Union's Area of Freedom,Security and Justice. The supranational directive, implemented by Italian LegislativeDecree no. 108 of 2017, has given rise to a construct of hybrid nature, inspired by theprinciple of mutual recognition, which maintains, at the same time, certain featurestypical of traditional mutual legal assistance, in an attempt to combine investigativeefficiency and protection of fundamental guarantees.In an underlying backdrop still characterized by the absence of harmonization ofnational procedural and evidentiary rules, the mechanism for adducing evidence in aforeign country revolves around the principle of proportionality, which in turn takesshape in the context of a balancing judgement - to be conducted in the actual case andtaking into consideration the specificities of such case - between the needs related tothe detection of crime and the sacrifices imposed on the rights of the personsinvolved, for various reasons, in the procedures aimed at issuing and executing therelevant order. This doctoral thesis intends to provide a comprehensive analysis of theEuropean Investigation Order, beginning with its legal framework, for the purposesof highlighting the main problems that have emerged in its early-stage enforcementand of identifying solutions capable of shorten the gap between theory and practice.To this end, a large space is firstly dedicated to the analysis of the early case-lawrendered by the Court of Justice and by the Italian Court of Cassation on this theme,which reveals the overall tendency to prefer purposes of investigatory efficiency tothe detriment of defense rights; secondly, this thesis critically evaluates somepractical cases selected at the Public Prosecutor's Office of Milan and Monza.
L'ORDINE EUROPEO D'INDAGINE PENALE: DISCIPLINA NORMATIVA E PRIME ESPERIENZE APPLICATIVE / C. De Luca ; tutor: L. Camaldo ; coordinatore: C. Luzzati. Dipartimento di Scienze Giuridiche Cesare Beccaria, 2022 May 17. 34. ciclo, Anno Accademico 2021.
IN VITRO AND IN VIVO APPROACHES TO STUDY OXIDATIVE STRESS, ANEMIA AND DYSBIOSIS IN CHRONIC KIDNEY DISEASE
CKD is diagnosed when there’s a decreased kidney function shown by a GFR less than 60 ml / min (established for a reference man with 1.73 m² body surface area), or markers of kidney damage, or both, of at least 3 months duration. The severity of complications increases in parallel with the GFR decline. We focused on three comorbidities extremely common in CKD patients: oxidative stress and inflammation; anemia and dysbiosis. We investigated these CKD comorbidities both with in vitro and in vivo approaches. More in detail, regarding in vivo studies, we measured oxidative stress biomarkers in a population of ESRD patients before and after the hemodialysis treatment, comparing the results with a population of healthy subjects; we evaluated oxidative stress biomarkers in the plasma of HD patients before, during and after two type of iron treatments (intravenous and sucrosomial iron). Regarding in vitro experiments, we focused on two uremic toxins, urea and indoxyl sulphate, and we evaluated their effects on a human endothelial cell line (Human Microvascular Endothelial Cells 1, HMEC-1).
IN VITRO AND IN VIVO APPROACHES TO STUDY OXIDATIVE STRESS, ANEMIA AND DYSBIOSIS IN CHRONIC KIDNEY DISEASE / E. Astori ; tutor: I. Dalle Donne ; commissario interno: N. Gagliano ; commissario esterno: S. Moreno ; commissario esterno: A. E. Colombo ; head pf phd course: F. Ficetola. Università degli Studi di Milano, 2021 Feb 23. 33. ciclo, Anno Accademico 2020. [10.13130/astori-emanuela_phd2021-02-23].
I PIANI E I PROGRAMMI NEL DIRITTO AMMINISTRATIVO. UNA TEORICA GENERALE, NELLA PROSPETTIVA DELLA DIGITALIZZAZIONE DELLA PUBBLICA AMMINISTRAZIONE
The research focuses on studying the planning procedures within administrative law, with a particular emphasis on the digitalization of public administration. The rationale for the investigation is based on the observation that in recent years, numerous and in-depth studies have been conducted on the digitalization of administrative activities, management of public administration databases, transformation of administrative functions, especially those related to knowledge, determined by technological advancements, and the enhancement of public information assets. Reflections on cybersecurity have also been prevalent, addressing a specific traditional area of public administration activity – public security – and studying its different and new object constituted by a new domain of action, no longer territorial but digital. Many of these studies have also been strongly influenced by the proliferation of supranational regulatory frameworks aimed at establishing a single digital market. All the aforementioned studies demonstrate a strong awareness that the digitalization process is well underway, and that public organization is already showing significant changes that can be examined from a purely legal perspective. Indeed, there is frequent attention to the use of new technologies in the activities of public administrations to improve efficiency and ensure better service to the community, in terms of good governance and impartiality in providing non-discriminatory services. However, it is believed that similar attention has not been given to the processes of political-administrative direction that govern these organizational processes. Assuming that planning processes are, among administrative procedures, those that directly contribute to political-administrative direction, this study aims to fillthis gap by studying the Three-Year Plans for Informatics developed by AgID within the framework of planning procedures, considered a privileged venue for forming political-administrative direction in digital matters.In the first part of the thesis, planning procedures are studied as administrative acts with general content. This study is necessary for two main reasons. Firstly, on an institutional study level, it meets the need to frame these procedures within the most shared elaborations regarding the classification of administrative procedures, thus elucidating their main formal characteristics. Secondly, from a critical perspective, this initial analysis highlights the problem arising from the legal nature of such procedures, which previously only intersected with the realm of normativity – generating contrasts, and relative positions, regarding the criteria to distinguish regulatory acts (regulations) from administrative acts with general content (plans and programs). Then, at a later stage, this analysis is enriched by the entry into the legal system of new subjects, particularly independent administrative authorities, new regulatory acts, their regulations, and new types of norms, technical ones, characterized by different legal efficacy. In addition to an institutional framework, the first chapter allows us to exclude the usefulness of resorting to formalistic elements to isolate the specificity of general administrative acts and highlights the need to interpret them as typical discretionary instruments of the activity of forming and implementing political-administrative direction.The second chapter, building on this acquisition, moves on to examine programming experiences as models of public intervention in the economy and therefore planning procedures suitable for the formation of a global economic policy direction, and then a sectoral one. In particular, global programming experiences are analyzed, followed by sectoral programming experiences that develop in the mature regionalism phase. In this chapter, which proposes a historical reconstruction of the transition from one
I PIANI E I PROGRAMMI NEL DIRITTO AMMINISTRATIVO. UNA TEORICA GENERALE, NELLA PROSPETTIVA DELLA DIGITALIZZAZIONE DELLA PUBBLICA AMMINISTRAZIONE / M. Palma ; tutor: G. Bottino ; coordinatore: F. Biondi. Dipartimento di Scienze Sociali e Politiche, 2024 Jul 09. 36. ciclo, Anno Accademico 2022/2023.
AN INTEGRATED PROTEOMIC AND ANALYTICAL APPROACH FOR ELUCIDATING THE MECHANISM OF ACTION OF HISTIDINE DIPEPTIDES AND SYNTHETIC DERIVATES
β-alanil-L-histidine (i.e. carnosine) is an endogenous peptide that have beenextensively characterized for a number of in vitro properties (i.e. metal chelating,antioxidant, reactive carbonyl species quenching). Several clinical trials highlightedthe potential benefits of carnosine in the treatment of oxidative stress-baseddiseases, although the in vivo mechanism of action is not known, yet. The researchproject herein tries to expand upon the in vivo mechanism of action of carnosine.New analytical methods have been developed by means of liquid chromatography– tandem mass spectrometry for the quantification of histidine dipeptides, theirderivatives, and the adducts formed with reactive carbonyl species intobiospecimens.A first step was the implementation of hydrophilic interaction chromatography toskip some sample preparation steps and to reduce the chance of systematic errors.The method allowed the quantification of carnosine and carnosinol (a carnosinederivative stable to carnosinase) in biospecimens. Carnosinol tissue distribution inanimal models of metabolic syndrome was determined and carnosinol-acroleinadduct was detected for the first time in liver matrices. This finding experimentallyconfirmed the reactive carbonyl species (RCS quenching activity of histidinedipeptides and derivatives in vivo. However, the metabolic instability of carnosinolHNE adduct was proved and such an evidence requires further studies aiming at understanding the metabolic fate of RCS-adducts to characterize their disposal.Subsequently, a new method for the measurement of carnosine hydrolysis in serumwas developed as well. Human serum carnosinase has been identified as theenzyme responsible for such an activity. Compared to other published assays, themethod employs a direct detection of the substrate and the use of less sample.Competition experiments with either natural derivatives or other molecules wereset to identify hit compounds acting as carnosinase inhibitors. The collected datawere shared with computational chemists who identified putative hit compoundsvia docking, virtual screening, and molecular dynamic approaches.Furthermore, a novel carnosine mechanism of action was studied starting from theevidence that carnosine can prevent the formation of protein adducts with 3,4-dihydroxyphenylglycolaldehyde (DOPEGAL) (i.e. an aldehyde intermediate ofnorepinephrine metabolism). This could be relevant for the in vivo mode of actionof carnosine since DOPEGAL can accumulate in cells because of oxidative stress andas it covalently binds proteins, it can alter their structures and functions.Carnosine quenching activity via the formation of an Amadori product withDOPEGAL was determined in vitro and in cell lysates producing DOPEGAL fromenzymatic transformation of norepinephrine. Future studies should be done tocharacterize the metabolic stability of the adduct and its formation in biospecimensas potential biomarker of norepinephrine toxicity.Finally, the project included proteomics studies on human umbilical vein cells(HUVECs) to assess the impact of carnosine and carnosinol on protein expression.It is widely recognized that drugs exert their pharmacological effects also by analteration of biological pathways by modifying protein expression. Carnosine andcarnosinol have little or no impact on protein expression as detectable on proteomeor secretome of healthy endothelial cells. In the future the impact on pathologicalcells should be carried out as well. These data support the hypothesis of a lowtoxicity for these molecules, making them suitable candidates for a chronicadministration.Although a lot of questions are still unanswered, these data have given new insightsin the mechanism of action of carnosine and in the discovery of molecules actingeither as carnosine-like compounds or as carnosinase inhibitors.
AN INTEGRATED PROTEOMIC AND ANALYTICAL APPROACH FOR ELUCIDATING THE MECHANISM OF ACTION OF HISTIDINE DIPEPTIDES AND SYNTHETIC DERIVATES / E. Gilardoni ; tutor: L. Regazzoni, G. Aldini ; coordinatore: G. Aldini. Dipartimento di Scienze Farmaceutiche, 2021 Jan 13. 33. ciclo, Anno Accademico 2020. [10.13130/gilardoni-ettore_phd2021-01-13].
UP150 PROACTIVE OFFICE: THE OFFICE THAT EDUCATES ON EMPLOYEE CARE AND WELL-BEING THROUGH PHYSICAL EXERCISE FOR HEALTH
"Systems thinking is a new area of knowledge that considers and integrates all the elements that must be considered to address complex public health problems, such as non-communicable disease prevention policy. Any intervention aimed at successfully preventing risk factors such as those caused by insufficient physical activity must be based on specific theories and scientific evidence and develop through active involvement in meaningful actions and interactions that consider the cultural and social context. For these reasons, the present study's structure is based on social theories contextualized in the sedentary working environment of desk workers to introduce a new company vision. In this vision, the working environment, where employees spend much time, is intended not only as a place for productivity but also a care and an educational environment to prevent a sedentary lifestyle. The aim of this study is to check with scientific and experimental experience how the traditional paradigm, stating that ""physical exercise intended as an active break out of context is opposed to useful and effortful work and represents a waste of time"", is no longer valid. The main idea is to incorporate physical exercise into the usual workflow to help employees increase their motivation to exercise and the amount of physical activity they do daily, with the aim of improving their health, physical literacy, and working wellness. Following the action research's vision, the present research project has been demonstrated to modify the local companies' welfare policies in which we proposed the intervention. Furthermore, the research structure following the design-based research paradigms has constituted an efficient road map to assess the effectiveness of the results of the proposed interventions. Finally, following the rationale of the system thinking approach, the research analyzed a wide range of variables (and their relationships) that can influence the analyzed reality to promote healthy lifestyles and long-term movement culture."
UP150 PROACTIVE OFFICE: THE OFFICE THAT EDUCATES ON EMPLOYEE CARE AND WELL-BEING THROUGH PHYSICAL EXERCISE FOR HEALTH / G. Signorini ; tutor: P.L. Invernizzi ; coordinator: C. Gozzoli. - Università Cattolica del Sacro Cuore, via Carducci 28 aula C014. Dipartimento di Scienze Biomediche per la Salute, 2025 Mar 26. 37. ciclo, Anno Accademico 2024/2025.
A COMPARISON BETWEEN GEOMETRIC QUASI-FUNCTORS AND FOURIER-MUKAI FUNCTORS
In this PhD thesis I have delt with the relationship between the most important class of functors between derived categories, i.e. Fourier-Mukai functors, and quasi-functors which are the morphisms in the localization Hqe of the category of dg categories with respect to quasi-equivalences.To be more precise: let X and Y be two smooth and proper schemes over a field. I have defined an explicit bijection between the isomorphism class of the triangulated category of perfect complexes over X×Y and the set of morphism in Hqe between two (fixed) dg enhancemenst of the categoriesof perfect complexes over X and over Y , respectively. Moreover, I have showed that this bijection associates to the dg lift of a Fourier-Mukai functor the isomorphism class of its kernel, giving a positive answer to a conjecture of Toën.
A COMPARISON BETWEEN GEOMETRIC QUASI-FUNCTORS AND FOURIER-MUKAI FUNCTORS / L. Dasti ; tutor: P. Stellari ; correlatore: A. Canonaco ; coordinatore: V. Mastropietro. Università degli Studi di Milano, 2023 Jan 26. 34. ciclo, Anno Accademico 2021.
DIETARY PATTERNS:NUTRITION, HEALTH, AND THE ENVIRONMENT
Growing evidence stresses the critical role of nutrition and dietary patterns in maintaining human health and preventing several chronic non-communicable diseases and certain types of cancer. In particular during the last decades the Mediterranean Diet acquired more and more importance. Defined a nutritional model shared by the population surrounding the Mediterranean basin around the fifties by Ancel Keys, is nowadays composed by olive oil, cereals, seasonal fruit and vegetables, legumes and nuts, moderate-to-high fish intake, seafood, fermented dairy products, poultry and eggs, together with a limited consumption of red and processed meats and sweets, and a moderate alcohol intake - when culturally allowed. Firstly, the adherence to the MD of a sample of pregnant women from Gran Canaria (Spain) and its possible implications in their newborn anthropometrics, is described. In second instance, 44 subgroup analyses within the scope of the PREDIMED study are presented. It consists of a large, parallel group, multicentre, randomized, controlled, single-blind, nutritional-intervention trial, conducted in Spain in 2003-2011 on 7447 participants, originally designed to assess the effects of the MD on the primary prevention of cardiovascular disease.Of note, beyond the largely demonstrated beneficial effects of the MD on human health, it is important to observe how food consumption, production and distribution patterns represent one of the main causes of ecological pressure on the environment. If so, global population’s dietary patterns link worldwide human health with the environmental sustainability and, considering population’s growing projections, this becomes an issue of major importance. In this perspective, we propose a new graphical representation of the MD pyramid, revisited according to the environmental aspects of food chain and represented with a third dimension. The overall value of the MD on the environmental, social, cultural, economic and nutritional/health dimensions, makes it a model that is considered as intangible heritage of humanity, but with added environmental value. The virtues of the MD and particularly its value for environmental preservation and protection need to be efficiently communicated and applied in an integrated manner at multiple levels. In this way diet will have a potential to nourish human beings and preserve our environment.
DIETARY PATTERNS:NUTRITION, HEALTH, AND THE ENVIRONMENT / L. Tomaino ; tutor: C. La Vecchia ; supervisor:Serra Majem, Lluis ; coordinator: C. La Vecchia. Dipartimento di Scienze Cliniche e di Comunità, 2021 Mar 10. 34. ciclo, Anno Accademico 2020. [10.13130/tomaino-laura_phd2021-03-10].
MOLECULAR MECHANISMS UNDERLYING CLINICAL BEHAVIOR OF NEUROENDOCRINE TUMORS: STUDY OF NEW POSSIBLE BIOMARKERS PREDICTABLE OF PHARMACOLOGICAL RESISTANCE AND TARGETS
Neuroendocrine tumors (NETs) represent a heterogeneous group of malignances arising from the diffuse neuroendocrine system whose incidence is rising over the years. Due to their complexity and unusual features, first-line therapeutical approach is represented by surgical resection, even if it is not completely resolutive in most of cases, due to recurrence and metastasis accompanied by pharmacological resistance. In this scenario, the lack of effective pharmacological approaches is making hard the treatment of these pathologies. Within the pharmacological context, lot of attention was given to somatostatin and dopamine system, because of prevalent expression of somatostatin receptors (SSTs) and dopamine receptor type 2 (DRD2) in about all NETs that represented, over the years, the main targets for treatment with somatostatin analogs (SSAs) and dopamine agonists (DAs) for these pathologies.Nevertheless, the molecular mechanisms underline pharmacological resistance and the development of novel biomarkers useful to identify patients that may benefit of specific treatments are steel needed, together with the development of novel in vitro pre-clinical model better resembling human tumor behavior. This thesis aimed to investigate possible biomarkers underlining pharmacological resistance and predictable of SSAs/DAs responsiveness in NETs, more specifically focusing on G-proteins and β-arrestins DRD2-mediated signaling pathways in PitNETs and on somatostatin/cortistatin system in novel PanNENs cell models.Briefly, β-arrestin 2 has been highlighted as novel potential biomarker of pharmacological responsiveness to DAs, and that combined therapy with DAs avoids everolimus escape feed-back in PitNETs. The role of -arrestin 2 in regulating both AKT phosphorylation and cell proliferation in tumoral lactotrophs and NF-PitNETs was demonstrated, elucidating a new molecular mechanism that contributes to confer sensitivity or resistance to DAs in PitNETs. In tumoral lactotrophs and NF-PitNETs, the lack of -arrestin 2 prevents the inhibitory effect of DRD2 on AKT pathway activation with a consequent resistance to the antimitotic action of DAs. In accordance, it was showed that cabergoline improved everolimus efficacy by blocking AKT upstream reactivation in NF-PitNETs and tumoral lactotrophs. In fact, the combined treatment with cabergoline strongly reduced the increase of AKT activity, correlating with its antimitotic effect. The role of -arrestin 2 has been highlighted in mediating cabergoline inhibition of both p-AKT activity and cells proliferation after everolimus cotreatment: the lack of β-arrestin 2 reverted the antimitotic effect induced by the combined treatment. Moreover, the cytoskeleton actin binding protein filamin-A (FLNA) emerged as an important player in the regulation of PitNETs drug responsiveness and invasiveness. At this regard our data showed that activation of the cAMP pathway and DRD2 agonist regulated FLNA activity by increasing or decreasing, its phosphorylation, respectively. FLNA phosphorylation has been identified as potential biomarker for tumor responsiveness to DAs, in DRD2 expressing PitNETs, since it participates in mediating DRD2 intracellular signalling, additionally turning out that FLNA phosphorylation prevented DRD2 signaling in PRL- and ACTH-secreting PitNETs.At last, a potential of novel pharmacological compounds of the SS/CST family exerted antiproliferative effects in new human resembling PanNENs models.To sum up, this work identified the role of specific molecular mechanism underlying pharmacological resistance in PitNETs, paving the way for possible combined approaches to overcome drug resistance and the pilot study on novel PanNENs cell lines strongly suggests that testing new pharmacological compounds of the SS/CST family can lead to an innovative discovery to expand the therapeutic arsenal to treat PanNENs and set novel approaches with a personalized perspective.
MOLECULAR MECHANISMS UNDERLYING CLINICAL BEHAVIOR OF NEUROENDOCRINE TUMORS: STUDY OF NEW POSSIBLE BIOMARKERS PREDICTABLE OF PHARMACOLOGICAL RESISTANCE AND TARGETS / F. Mangili ; tutor: G. Mantovani Giovanna, MD, PhD ; coordinatore: C. Sforza Chiarella, MD, PhD. Dipartimento di Scienze Cliniche e di Comunità, 2023 Apr 18. 35. ciclo, Anno Accademico 2022.
VOCES QUE RESISTEN:EL TESTIMONIO CHILENO DE LA DICTADURA ESCRITO POR EX PRESAS POLÍTICAS
"My doctoral thesis analyses testimonial literature written by former political prisoners between 1973 and 2017 and its title is ""Voces que resisten: el testimonio chileno de la dictadura escrito por ex presas politicas"". It offers a thematical analysis of a corpus of thirteen texts and is developed in three chapters. The first chapter traces the history of testimonial literature and the Latin American theorical debate about it, with specific attention to Chilean context. I divided Chilean testimonial literature chronologically in three stages, in order to tie it to its historical background, as similarly done by Peris Blanes (Cfr. 2002, 2005, 2008), Moulian (Crf. 1997) and Richard (Crf. 2001, 2010, 2017). Thus it is read first as a form of resistance to the dictatorship, then within reconciliation policies and at last as realized in the contemporary world. In addition to this theoretical and historical introduction, the first chapter presents also the examined works, placing each one into the threefold historical context previously observed. Narrative differences between works are then taken in account: the narrative construction of the heroic woman and / or widow immolated to the cause (typical of the first stage and for this reason we speak of 'classical' texts); the one close to confession (which falls within the historical period of reconciliation); and, finally, contemporary texts (published between 2000 and 2017 and presenting diversified narrative modes). In the second chapter, the methodology is explained, and the analysis is carried out. It performs a gender study which takes on the feminine nature of the writers, by enlightening typically female themes such as the difficulty of reconciling political militancy with womanhood, motherhood and sexual violence - often a weapon of state violence during political imprisonment. The chapter also focuses on the thematic novelties of contemporary texts, the modification and evolution of which can bring to reopening the debate on the dictatorial past and bringing back 'forgotten' themes (such as militancy, the concept and importance of friendship networks). Finally, in the third chapter, the analysis is carried out with the usual diachronic approach and through three proposed thematic axes: resistance, resilience and solidarity. Testimonial literature is considered as an act of resistance itself since ""el testimonio es una forma de organizar la resistencia a las violaciones de los derechos humanos [...] escribir sobre la revolución es hacer la revolución"" (Nofal 2002: 34 -35). Writing is a way of denouncing the horror of dictatorial violence and telling how resistance has developed in different forms, such as the clandestine struggle, the reconfiguration of a dissident organization in exile and the return to continue the fight. For this reason, within the texts is looked for all those acts that can be linked to the resistance, that is understood also as mobilization of forces - even hidden and ""lateral"" - which undermine central power (Calveiro 2003: 92). Resilience means to outline gestures born/blossomed in a profoundly violent context, such as detention and torture centres, and that count for the human capacity to adapt to extreme situations without however losing their dignity and remaining ""human"". In this part, all those artistic and cultural productions born as a form of elevation and rebirth are also examined. The third theme, called ""solidarity"", highlights all the forms of solidarity, of sisterhood and brotherhood, which allowed the survival of friendship bonds between militants, even in extremely difficult situations. Clearly, these three themes dialogue and intertwine one another, recreating the social and political community these authors were part of. Therefore, the conclusions clarify how the community constraint has remained strong, despite the passage of time and the dictatorial attempt to annihilate all the di"
VOCES QUE RESISTEN:EL TESTIMONIO CHILENO DE LA DICTADURA ESCRITO POR EX PRESAS POLÍTICAS / S. Cappellini ; direttore di tesi estero: C. Pizarro Cortés : direttore di tesi italiano: L. Scarabelli ; coordinatore del dottorato: M. V. Calvi. Università degli Studi di Milano, 2020 Feb 21. 32. ciclo, Anno Accademico 2019. [10.13130/cappellini-serena_phd2020-02-21].
IL CONTROVERSO INQUADRAMENTO DELL'OPPOSIZIONE ALLO STATO PASSIVO FALLIMENTARE
"The purpose of the study conducted was to identify the rules governing the procedure for opposing the bankruptcy statement of liabilities, expressly provided for in Articles 98-99 of the bankruptcy law, as a possible remedy against the decree of enforceability of the statement of liabilities issued by the delegated judge.The issue under consideration was of particular interest because, first of all, it made it possible to analyse the profound changes that the proceedings in question have undergone since the issue of R. D. 267/1942, followed by the reform carried out with D.lgs 5/2006 corrected with D.lgs 169/2007 and culminating in the Dlgs 14/2019. Secondly, it allowed a critical evaluation of the troubled legislative process mentioned, which did not always comply with the intention of creating a complete set of rules suitable to speed up the process.Therefore, the beginning of the dissertation is constituted by a comparison between the discipline of the verification of credits as outlined by the legislator of 1942 and the modifications introduced in the two-year period 2006-2007. First of all, the first phase of the judge's activity was examined, which, in its original structure, on the border between the administrative and the jurisdictional nature, was dominated by the summary nature and by the inquisitorial principle, and was linked to a singular breadth of powers granted to the judge in order to simplify the procedure. The second phase, on the other hand, unlike the first, was subject to the discipline of the ordinary rite of cognition, although introduced with an appeal, instructed by the same delegated judge, a rite in which the parties had to respect the endo-procedural terms provided for by art. 183 ss c.p.c.. At that time, the nature of the appeal was recognised as having devolutive effect: in other words, the provisions of Article 345 c.p.c. had to be applied.With the novel, the legislator wanted to assign to the delegated judge and the receiver a profoundly different role from the past, outlining the procedure of verification of claims as a procedure that is of a contentious jurisdictional nature, albeit with summary knowledge, and that is based on the principle of the application and the exception, in which the receiver assumes the role of party and the judge pronounces according to the rules and in compliance with the rules of evidence. The procedure relating to ""appeals"", on the other hand, has been maintained as a judgment on the merits with full knowledge of the facts, even though it is characterised by a single rite for the treatment of the three different means of appeal. Moreover, the discipline of this rite is no longer automatically classifiable within the scope of the norms that mark the ordinary process of cognition, since, with the reform, the references to the code of ordinary rite no longer exist.Having defined the essential features of the reforms mentioned, the second and third chapters focus on the search for the legal nature of the opposition procedure. And so, first of all, we have adhered to the conviction that the remedy de quo can be counted within the category of the means of appeal, not being able to consider it as a simple continuation, with full knowledge, of the summary phase of verification. This classification is not without consequences on the level of discipline, which has made it necessary to study the devolving effect and the prohibition of ius novorum, and then to proceed to a comparison with the model described in Articles. 339 ss. C.p.c., which concludes with the impossibility of overlapping the two remedies. Specifically, the different attitudes of the devolving phenomenon, of the exceptions, of the evidence that can be deduced, of the prohibition of reformatio in peius and, above all, of the possibility of lodging incidental appeals were analysed.Having identified the characteristics of the opposition as a model of appeal that cannot be equated with the judgment on"
IL CONTROVERSO INQUADRAMENTO DELL'OPPOSIZIONE ALLO STATO PASSIVO FALLIMENTARE / F. Carletti ; tutor: S. Vincre' ; coordinatrice: M. T. Carinci. DIPARTIMENTO DI DIRITTO PUBBLICO ITALIANO E SOVRANAZIONALE, 2020 Feb 12. 32. ciclo, Anno Accademico 2019. [10.13130/carletti-federico_phd2020-02-12].
DUALITIES AND EXACT RESULTS IN SUPERSYMMETRIC QFTS
In this thesis we analyze supersymmetric QFTs in three and four spacetime dimensions, with the goal of studying the infrared behavior of these theories.In Part I we analyze various dualities involving 3d N=2 theories with two tensor or adjoint fields and D-type superpotential. New dualities are discovered and we show relations between those and other 3d and 4d dualities. We also study 3d N=2 SQCD theories with orthogonal or symplectic gauge groups and monopole superpotential that exhibit s-confinement. These results are checked through the computation of the three-sphere partition function and via confinement/deconfinement procedures.In Part II we study exactly marginal deformations of 4d SCFTs and identify instances of theories that flow to different points of the same conformal manifold, a phenomenon denoted as conformal duality. We study a series of 4d N=1 theories engineered by a stack of D3-branes in Type IIB probing a Calabi-Yau toric singularity in the presence of an orientifold and build infinite families of pairs of conformally dual theories.In Part III we classify the 1-form symmetry groups of N=3 S-fold SCFTs as well as their global structures by analyzing the maximal refinements of their charge lattices. We study the charge lattices of S-folds and exceptional S-fold SCFTs of type E. We find that all but one of the exceptional S-folds do not admit a charge lattice consistent with the moduli space and must be discrete gaugings of free theories. We support this claim by developing constraints on the stratification of Coulomb branch and bounds for the 1-form symmetry group for a class of N=2 SCFTs.
DUALITIES AND EXACT RESULTS IN SUPERSYMMETRIC QFTS / S. Rota ; supervisore: A. Amariti ; coordinatore: M. Paris, R. Vecchi. - Via Celoria, 16 Milano. Università degli Studi di Milano, 2024. 36. ciclo, Anno Accademico 2023.
IMMUNOGENIC CELL DEATH AS A NEW MECHANISM OF ACTION OF THE CHEMOTHERAPEUTIC DRUG MITOMYCIN C IN BLADDER CANCER
Despite having been used for more than a century, the exact mechanisms of action, of resistance and the best treatment schedule of most chemotherapeutic agents remain elusive. Mitomycin C (MMC) is the gold standard adjuvant treatment for bladder cancer. However, it is effective only in a proportion of patients, suggesting that, aside from cytotoxicity, other mechanisms could be involved in mediating the success or failure of treatment. We hypothesized that MMC might induce immunogenic cell death (ICD), leading to an antitumor immune response. Here, we describe that MMC fosters ICD via the exposure of damage signals, increased phagocytosis by dendritic cells (DCs) and in vivo tumor protection. MMC-induced ICD relies on the cytoplasmic release of mitochondrial DNA that activates the inflammasome for efficient IL-1β secretion that promotes DC maturation. We found the ICD resistant cancer cells fail to generate an inflammatory microenvironment and display mitochondria dysfunction related to low expression of mitochondrial Complex I of the respiratory chain, which is associated with drug resistance in bladder cancer patients. The identification of ICD as a novel immune-related mechanism of action of MMC provides opportunities to optimize bladder cancer management and identify ICD-related biomarkers of treatment efficacy.
IMMUNOGENIC CELL DEATH AS A NEW MECHANISM OF ACTION OF THE CHEMOTHERAPEUTIC DRUG MITOMYCIN C IN BLADDER CANCER / B. Oresta ; supervisor: M. Rescigno. DIPARTIMENTO DI SCIENZE DELLA SALUTE, 2020 Jan 28. 31. ciclo, Anno Accademico 2019. [10.13130/oresta-bianca_phd2020-01-28].
SYNTHETIC-LETHAL PHARMACOLOGICAL INTERACTIONS IN MYC-DRIVEN LYMPHOMAS: A ROLE FOR GLUCOCORTICOIDS IN TARGETED THERAPY
Diffuse large B-cell lymphoma (DLBCL), the most common non-Hodgkin lymphoma, includes aggressive subtypes that remain difficult to treat. One of the molecular features associated with poor prognosis is the presence of chromosomal translocations that activate the MYC oncogene, in particular when associated with concurrent activation of BCL2, which leads to an aggressive disease subtype commonly known as double-hit lymphomas (DHL). Since direct targeting of MYC remains challenging, we sought to unravel synthetic-lethal vulnerabilities as alternative therapeutic options against MYC-driven tumors. Toward this aim, we performed an unbiased high-throughput pharmacological screen in a murine B-lymphoid cell line with conditional MYC activation and identified a set of compounds that preferential kill MYC-overexpressing cells. Unexpectedly, these included glucocorticoids (GCs) with Budesonide scoring as the top candidate. Targeted experiments confirmed that MYC also sensitizes B-cells to another GC, Prednisolone, the active metabolite of Prednisone, itself an integral component of the frontline therapeutic regimen R-CHOP. Functional assays revealed that MYC activation markedly sensitizes B cells to GC-induced apoptosis through the intrinsic mitochondrial pathway. Transcriptomic profiling confirmed that MYC differentially modulates the transcriptional response to GCs, reinforcing the activation of stress and apoptotic programs while attenuating the repression of proliferative and metabolic genes, thus providing a mechanistic basis for MYC-specific cytotoxicity. Guided by these findings, we uncovered a strong synergy between GCs and the selective BCL2 inhibitor Venetoclax (ABT199) in killing a human DHL cell line, providing a rationale for combining these drugs against this aggressive lymphoma subtype. Additional combinatorial experiments showed that Prednisolone contributes to the cytotoxicity of the pharmacological CHOP backbone in MYC-overexpressing B-cells. Collectively, our data identify GCs as potent MYC-synthetic lethal agents, mechanistically linked to MYC-driven remodeling of GC-regulated transcription, revealing an unexpected mechanism of action for these long-standing therapeutic agents.
SYNTHETIC-LETHAL PHARMACOLOGICAL INTERACTIONS IN MYC-DRIVEN LYMPHOMAS: A ROLE FOR GLUCOCORTICOIDS IN TARGETED THERAPY / A. Ghsein ; tutor: B. Amati ; co-tutor: G. Donati ; internal advisor: S. Minucci ; external advisor: L. Soucek ; coordinator: D. Pasini. Dipartimento di Oncologia ed Emato-Oncologia, 2025 Dec 17. 37. ciclo, Anno Accademico 2024/2025.
PHENOTYPIC AND MOLECULAR CHARACTERIZATION OF A NOVEL P63ALPHA;KO MOUSE MODEL
The transcription factor p63, a p53 gene family member, plays a crucial role in embryonic development of stratified epithelia where it can function either as an activator or a repressor. In the epidermis, p63 is a master regulator of epidermal gene expression and is crucial for cell proliferation, cell adhesion and stratification. Here we describe a new p63α-/- mouse model in which we delete exon 13 leading to the expression of the p63β isoform. Newborn p63α-/- mice die soon after birth and are characterised by cleft palate and digit malformations but exhibit normal skin. Our study indicates that p63α and p63β are not interchangeable in palate and limb development during embryogenesis, whereas expression of either one allows normal skin development, laying the foundation for a better understanding of the contribution of each p63 isoforms to mouse development. Compared to previously generated p63 knock-out mice, morphological and molecular impairment in p63α-/- model are mainly caused by alterations in the expression of genes involved in cell-cell and cell-matrix interactions. Taken together, these novel findings shed light on the function of both the alpha and the beta isoforms, and indicate that p63α is specifically required for limb and palate development, whereas p63β can replace p63α in skin development and in the adulthood.
PHENOTYPIC AND MOLECULAR CHARACTERIZATION OF A NOVEL P63ALPHA;KO MOUSE MODEL / G. Urciuoli ; supervisor: C. MISSERO ; internal supervisor: B FRANCO ; external supervisor: M. MIKKOLA. Università degli Studi di Milano, 2023 Mar 02. 34. ciclo, Anno Accademico 2022.
A SYSTEM OF SUPERLINEAR ELLIPTIC EQUATIONS IN A CYLINDER
This dissertation is primarily concerned with the study of existence of positive solutions of nonlinear elliptic boundary value problems. An important way to deal with the problem is the study of a priori estimates of positive solutions. We will adapt a classical idea which was introduced by Brezis and Turner and, together with a fixed point theorem, we will derive the existence result of a superlinear elliptic system which defined on a cylinder. First we present some of the history of this problem, along with the necessary mathematical background. We present the main technical tools: Hardy's inequality, regularity theory and maximum principle as well as the work of Brezis and Turner. They treated a general superlinear elliptic problem and obtained the existence of positive solutions for nonlinear term having an asymptotic growth $s^\gamma$ with $1<\gamma<\frac{n+1}{n-1}$. In the novel part we apply Brezis and Turner's technique to a specific elliptic system. We study the $L^p$ regularity theory, Hardy's inequality on a cylinder and with growth conditions imposed on the nonlinear term. In particular, we will find that the nonlinear term embeds into different $L^p$ spaces as the dimension $n$ varies. We point out that there is a regularizing effect in the system which leads to a larger exponent than the Brezis-Turner exponent.
A SYSTEM OF SUPERLINEAR ELLIPTIC EQUATIONS IN A CYLINDER / Y. Guo ; tutor: B. Ruf ; coordinator: V. Mastropietro. Università degli Studi di Milano, 2022 Jun 29. 34. ciclo, Anno Accademico 2021.
SOFT POLYMERIC NANOCOMPOSITES FOR ELECTROMECHANICAL CONVERSION
Soft electro-mechanical transducers are receiving a widespread interest for both sensing and energy harvesting applications. In the last decade the research on novel nanocomposites based on polymeric nanocomposites has been very active, and many materials with diverse and innovative compositions and architectures have been fabricated and reported. Nevertheless, there are still some critical aspects that affect this research field, in particular for what regards their characterization.Despite the widespread interest that these smart materials are attracting, the methods commonly used to characterize them are frequently qualitative and unreliable. Moreover, the description of the testing conditions is often incomplete, so that the data cannot be reproduced by other researchers, and the performances of the various materials cannot be compared. The knowledge of these properties is necessary for a complete understanding of the material working principle, for the prediction of its effective properties aimed at optimization purposes, and for the selection of the right material in device applications. In this framework the development of a quantitative and reliable measurement technique is crucial in order to to assess these materials figure of merit, their reliability and reproducibility.This thesis work is focused on the fabrication and characterization of materials with electro-mechanical conversion capabilities, for applications in both energy harvesting and pressure and strain sensing. Four different kinds of materials were developed and tested, three active materials based on piezoelectric BaTiO3 nanoparticles on different polymeric matrixes, that can be applied both as sensors and as energy harvesters, and a passive material based on a piezoresistive polymer/metal nanocomposite, which can be applied as a strain sensor. For the characterization of such nanocomposites, a custom experimental set-up to measure the piezoelectric coefficients in a wide frequency range was developed and validated.
SOFT POLYMERIC NANOCOMPOSITES FOR ELECTROMECHANICAL CONVERSION / S.m. Villa ; supervisor: P. Milani ; referee: R. Kay, L. Ricotti ; director of the thesis: M. Paris. Università degli Studi di Milano, 2022 May 19. 34. ciclo, Anno Accademico 2021.
THE INTERPLAY BETWEEN ENDOCRINE DISRUPTION AND GENETICS IN THE PHYSIOPATHOLOGY OF HUMAN NEURODEVELOPMENT: BRIDGING EPIDEMIOLOGY TO MOLECULAR INSIGHTS THROUGH BRAIN ORGANOIDS AND HIGH RESOLUTION OMIC MODELLING
Several epidemiological studies uncovered a strong link between pre- and postnatal exposure to EDCs and impaired development of cognitive functions, attention and social behaviors in children. Nonetheless, the molecular mechanisms underlying such outcomes are yet to be detailed and EDC-induced DNT (developmental neurotoxicity) remains largely uncovered by the current testing tools in regulatory use.We thus investigated the hormonal pathways underlying EDCs-induced DNT by taking advantage of human pluripotent stem cells (hiPSC)-derived cortical brain organoids (CBOs), a model that recapitulates salient species-specific features of human brain development in a 3D microenvironment characterized by stereospecific interconnections among different cell populations. After chronically exposing CBOs to 7 different hormones, their respective antagonists and their combinations, we leveraged bulk and single cell transcriptomic, as well as imaging-based quantification of brain cortex-specific developmental populations to characterize the transcriptional and functional alterations promoted by the activation and inhibition of each hormonal pathway and thereby defining the impact of their modulation on cortical organoids physiology. Subsequently, we performed a toxicological evaluation of a battery of EDCs, chronically exposing CBOs to such compounds, employing for each one a 4 log range of concentrations linked to those inferred by epidemiological studies to extensively define their impact on human cortex development.In parallel, to allow the experimental tractability of large cohorts of genotypes, fundamental to decipher the contribution of genetic variability in neurodevelopment, we explored and benchmarked genetic multiplexing approaches that allow to grow mosaic CBOs from multiple donors.
THE INTERPLAY BETWEEN ENDOCRINE DISRUPTION AND GENETICS IN THE PHYSIOPATHOLOGY OF HUMAN NEURODEVELOPMENT: BRIDGING EPIDEMIOLOGY TO MOLECULAR INSIGHTS THROUGH BRAIN ORGANOIDS AND HIGH RESOLUTION OMIC MODELLING / M.t. Rigoli ; added supervisor: N. Caporale ; internal advisor: F. Iorio ; tutor: G. Testa ; phd coordinator: S. Minucci. - human technopole. Dipartimento di Oncologia ed Emato-Oncologia, 2024 Apr 11. 35. ciclo, Anno Accademico 2023.
PERINATAL SEROTONERGIC MANIPULATION SHAPES LONG-TERM NEUROPSYCHIATRIC-LIKE PHENOTYPES IN RATS: MOLECULAR INSIGHTS ACROSS THE LIFESPAN.
Poor understanding of neuropsychiatric disorders often results in similar diagnoses for patients with different symptoms thus reducing medication effectiveness. Early research focused on the serotonergic system, highlighting serotonin (5-HT) playing a crucial role in the development of various central nervous system diseases, including neuropsychiatric disorders. Indeed, besides its role as a neurotransmitter, 5-HT is important as a trophic factor in brain development, affecting neural circuit formation and plasticity. Brain development relies on precise timing and deviations may disrupt brain function and lead to disorders manifesting later in life. Many animal models have explored 5-HT's role in neuropsychiatric conditions, especially with genetic approaches. These models often cause permanent changes and do not capture the system's dynamic nature or time-specific disruptions, complicating the interpretation of the results.On these bases, in my Ph.D. project, we took advantage of a pharmacological approach, that involves the manipulation of the serotonergic system in rats to study the effects of its dysregulation during specific early-life stages. The main aim of this study was to examine how perinatal serotonergic manipulation affects behavior and the underlying molecular changes, as well as its potential impact on vulnerability to neuropsychiatric conditions throughout life. Specifically, we exposed pregnant dams to the selective serotonin reuptake inhibitor fluoxetine (FLX) via drinking water, during gestation (from gestational day 0 to postnatal day (PND) 0) or breastfeeding (from PND 0 to weaning at PND21). As FLX crosses the placenta and enters breast milk, the offspring were also exposed to the drug during the prenatal or the postnatal periods respectively.We first evaluated the impact of FLX on birth rate and body weight. Our results showed that prenatal-FLX exposure reduced litter sizes and offspring body weight, though recovery occurred by weaning. Differently, the effect of postnatal-FLX exposure on weight gain lasts until adulthood despite FLX exposure ending at PND21.Three behavioral tests were used to evaluate different behaviors related to neuropsychiatric symptoms. Specifically, the sucrose preference test assessed hedonic behavior, the novel object recognition test evaluated cognitive performance, and the elevated plus maze test examined anxiety-related behavior. These tests, along with molecular analyses, were performed at various ages to determine when the effects of perinatal manipulations become evident.Behavioral assessments revealed that FLX exposure timing and the animals' sex influenced pathological-like traits, with males being more vulnerable to prenatal-FLX manifesting anhedonic-like behavior in adulthood and females more sensitive to early postnatal-FLX, showing cognitive deficits when adults. Molecular analyses indicated widespread changes in the blood, in the prefrontal cortex (PFC), and in the hippocampus (Hip), including immune system and transcriptional protein alterations. In addition, we found changes related specifically to each experimental group. In particular, our results showed disrupted steroid hormone responses in males of the prenatal-FLX groups and changes in the autophagic processes in females exposed to postnatal-FLX. We further investigated whether molecular changes are detectable before the manifestation of the behavioral phenotype. Focusing specifically on males exposed to prenatal-FLX, we found that the manipulation disrupted the molecular stress response in adolescents. Indeed, we observed prenatal-FLX altered the temporal profile of the hypothalamic-pituitary-adrenal (HPA) axis. Furthermore, FLX exposure prevented the typical post-acute stress increase in Nr4a1 and Dusp1 expression in the PFC, indicating broader disruptions in stress-related systems beyond the HPA axis. In the last part of my PhD project, we explored the hypothesis that perinatal seroto
PERINATAL SEROTONERGIC MANIPULATION SHAPES LONG-TERM NEUROPSYCHIATRIC-LIKE PHENOTYPES IN RATS: MOLECULAR INSIGHTS ACROSS THE LIFESPAN / M.t. Gallo ; tutor: F. Calabrese ; coordinatore: G. D. Norata. Dipartimento di Scienze Farmacologiche e Biomolecolari, 2024 Dec 18. 37. ciclo, Anno Accademico 2023/2024.
TRANSMEMBRANE CHLORIDE INTRACELLULAR CHANNEL 1 (TMCLIC1) PROTEIN IS A KEY REGULATOR OF COLORECTAL CANCER¿S PROLIFERATION AND METASTASIS
Colorectal cancer is the third leading cause of death in developed countries. Although the huge progresses reached in therapeutical approach, in the presence of metastasis colorectal cancer remains still uncurable. Ion channels have a prominent role in the acquirement of aggressive phenotype, and they are considered the ideal target to design an anticancer therapy. The main problem is that most of them are involved in normal physiological processes and treatments against their activity could result in high toxicity.From several years, our laboratory has focused its attention on the study of CLIC1 protein, considered a promisingcandidatefor a targeted anticancer adjuvant therapy. CLIC1 is a ubiquitous protein belonging to a partially unexplored protein family, the CLICs, that have the peculiar characteristic to be metamorphic. In normal conditions, CLIC1 protein is expressed by all cell types as a cytosolic monomer. Upon stress stimuli, monomers undergo a conformational change and translocate to the plasma membrane where increase chloride permeability. CLIC1 protein was found to be overexpressed in different pathological states, including cancer, but the mechanism in tumorigenesis is still unclear.Our laboratory has demonstrated that in glioblastoma stem cells tmCLIC1 is chronically expressed and here has a pivotal role in tumorigenesis and cancer development. CLIC1 mRNA overexpression was identified in colorectal cancer cells and is related to a poor prognosis. In this scenario, the main aim of the present work is to understand the role of tmCLIC1 in tumorigenesis, invasion, and migration potential of colorectal cancer.To this purpose, were chosen different human immortalized colorectal cancer cells at different stage of aggressiveness according to the Dukes’ classification; a method that categorizes tumor development based of the invasion of tissues and mucosa. Here, was investigated the level of expression of CLIC1 protein and the activity of its transmembrane form as an ion channel. Our results have demonstrated that tmCLIC1protein activity is proportional to the level of aggressiveness of cells. We evaluated the effect on cellular proliferation, migration and invasion of the specific inhibitor of tmCLIC1, IAA94, and a monoclonal antibody, designed in our laboratory and made to specifically recognized the NH2 terminal domain of CLIC1, exposed to extracellular space when the protein is docked in membrane. IAA94 and our antibody, now under license and named tmCLIC1omab, have demonstrated the same inhibitory proprieties. tmCLIC1omab could represent a good alternative for an adjuvant therapy to impair tmCLIC1 activity, considering the high toxicity in vivo of IAA94.Our results have demonstrated that the inhibition of tmCLIC1 activity results in a downregulation of proliferation in the most aggressive cell lines in 2D model, as well as in 3D models. Moreover, the migration and invasion potential of colorectal cancer cells is mainly abolished when tmCLIC1 is impaired. To demonstrate that the effects observed in the presence of tmCLIC1 inhibitors are due to a specific action of the drugs on the protein, and not for the presence of other off targets, we established stable cell lines in which CLIC1 was knocked down after a lentiviral infection. Results obtained in silenced cells have demonstrated the same phenotype of cells treated with IAA94 and tmCLIC1omab.The role of tmCLIC1 activity in tumorigenesis and cancer development was confirmed also in in vivo models. Experiments performed in Zebrafish embryos and immunodeficient mice have demonstrated that the silencing of CLIC1 results in the impairment of primary tumor development and formation of distal metastases. Experiments were also replicated in immunocompetent mice injected with murine colorectal cancer cells that have shown the same phenotype of human colorectal cancer cells used for all the previous trials. Our preli
TRANSMEMBRANE CHLORIDE INTRACELLULAR CHANNEL 1 (TMCLIC1) PROTEIN IS A KEY REGULATOR OF COLORECTAL CANCER¿S PROLIFERATION AND METASTASIS / F. Cianci ; tutor: M. Mazzanti. Dipartimento di Bioscienze, 2022 Mar 18. 34. ciclo, Anno Accademico 2021.
FROM DG-CATEGORIES OVER K TO K-LINEAR STABLE INFINITY-CATEGORIES
The purpose of this thesis is to prove that “idempotent-complete pretriangulated dg-categories over k are stable k-linear idempotent com-plete ∞-categories”. This result is a meeting point between algebraic topologyand algebraic geometry.In 2013 L. Cohn proved it in a preprint, and in 2016, he updated the preprint toits latest version.Since then the ∞-category theory and the spectral algebraic geometry theoryhave made progress: therefore, we think that a reproof can be useful in makethis result clearer for more mathematicians.The first four chapters are devoted to laying the groundwork for the next chapterwhere linear category theory can finally be presented. In the fifth chapter, wetackle for the first time the problem of defining the theory of k-linearization usingenriched ∞-category theory (Lurie has previously addressed this topic but witha different setting [1, Spectral Algebraic Geoemetry]). This chapter comprises three parts: the first two runalmost in parallel, while the third compares them. In the first part, we delveinto the theory of dg-categories over a fixed ring k, which serves as the algebraic-geometric setting for k-linearization. One significant result is the demonstrationthat M odHk-enriched categories are left Hk-module objects in CatSp∞ . This resultelucidates why the two parallel sections converge, prompting an additional sectionto present it in a categorical setting. This finding enables k-linear Morita theoryto be explored using nonlinear theory.In the second part, we investigate linearization from the viewpoint of spectralalgebraic geometry (or algebraic topology). We provide a conceptual overview oflinearization and define Morita theory for k-linear ∞-categories. In this part, wewill come to realize that there exists an incredible number of equivalent definitionsof k-linear ∞-categories, and the main outcome of this section is to demonstratetheir equality. In the final part of the fifth chapter, we achieve our goal and provethe main results. The thesis concludes with a brief chapter outlining possibledirections for future research following this work.
FROM DG-CATEGORIES OVER K TO K-LINEAR STABLE INFINITY-CATEGORIES / M. Doni ; tutor: P. Stellari. Dipartimento di Matematica Federigo Enriques, 2024. 36. ciclo
THE METHIONINE CYCLE AS A NOVEL THERAPEUTIC TARGET IN NPM1C DRIVEN ACUTE MYELOID LEUKEMIA
Acute myeloid leukemia (AML) is the most common leukemia in adults and its prognosis is usually poor. Patients are still treated with conventional chemotherapy, which induces disease cure in <40% of patients. The main culprit of therapy failure is the genomic and biological heterogeneity of the disease and the lack of specific targeting-approaches. NPM1 is the most frequently mutated gene in AMLs (~30% of cases). Though the role of the NPM1 mutation (NPM1c) in the maintenance of the leukemic state is well characterized, little is known about underlying molecular mechanisms of oncogenesis. Most notably, the mutation causes an aberrant cytoplasmic localization of the protein, which is otherwise a nucleus-cytoplasmic protein mainly localized in the nucleolus and in the nucleoplasm.Unpublished data generated in our lab have shown that ectopic expression of NPM1c in Murine Embryonic Fibroblasts (MEFs) leads to an unbalanced methionine-cycle with decreased ratio between S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH), key metabolites of the methionine cycle. Decreased SAM/SAH ratio is known to inhibit cellular methyltransferases activity. Consistently, the majority of NPM1c regulated genes (e.g. HoxA cluster) and enhancers in MEF cells result markedly hypomethylated and overexpressed or active, respectively.Here, we show that SAM exogenous supplementation in NPM1c-MEF cells increases DNA methylation at specific loci and reverts significantly expression and activity of NPM1c-targeted promoters and enhancers. We then investigated the effects of SAM supplementation in NPM1c-AML blasts, and demonstrated that SAM reverts the NPM1c transcriptional signature, promotes monocytic differentiation and cell death in vitro and inhibits AML growth in vivo, suggesting that restoration of a physiological SAM/SAH ratio reverts the malignant phenotype of NPM1c-AMLs. Using engineered AML cells that allow degradation of NPM1c or its ectopic expression, we demonstrated that NPM1c is directly responsible for: i) the cytoplasmic mis-localization of Methionine Adenosyltransferase 2A (MAT2A), the key enzyme that catalyzes SAM production, leading to SAM/SAH ratio unbalance; ii) the SAM/SAH ratio unbalancing; and iii) the sensitivity to SAM treatment. To explore alternative approaches to increase the SAM/SAH ratio, we used Metformin, known to activate adenosylhomocysteinase (SAHH) and reduce SAH levels, and showed that it synergizes with SAM, inhibits growth and promotes monocytic differentiation of NPM1c-AML growth in vitro.In summary, in this PhD project, we demonstrate that the SAM/SAH unbalance is at the basis of the NPM1c-dependency of AML blasts, thus paving the way for the therapeutic use of SAM as specific targeting approach in NPM1c-AMLs.
THE METHIONINE CYCLE AS A NOVEL THERAPEUTIC TARGET IN NPM1C DRIVEN ACUTE MYELOID LEUKEMIA / A. Polazzi ; tutor: P. G. Pelicci ; co-tutor: E. Colombo. Dipartimento di Oncologia ed Emato-Oncologia, 2023 Apr 13. 34. ciclo
PROMOTION AND MONITORING OF PHYSICAL ACTIVITY WITH A DIGITAL APP IN PATIENTS WITH IRRITABLE BOWEL SYNDROME: CLINICAL OUTCOMES AND FECAL MICROBIOTA ANALYSIS OF A RANDOMIZED CONTROLLED STUDY
Background and Aim: Physical activity (PA) improves irritable bowel syndrome (IBS) symptoms. Digital Apps may increase PA, but whether this intervention can improve symptoms in patients with IBS has not yet been investigated. Gut microbiota plays an important role in IBS. The aim of this randomized controlled trial was to evaluate the effects of PA prescription with the use of a mobile App.Methods: We enrolled 97 consecutive patients with IBS according to Roma IV criteria; 22 patients aged <18/>60 years or with comorbidities limiting PA were excluded. Patients were randomized to two intervention groups: 1) (Not APP) PA prescription with a 10 min counselling or 2) (APP) PA prescription with the indication to use the mobile App Google Fit after a brief explanation. Patients were evaluated at the first visit (T0) and after 3 months (T3). The severity of IBS symptoms (primary endpoint) was assessed by the Irritable Bowel Severity Scoring System (IBS-SSS). The Structured Assessment of Gastrointestinal Symptoms Scale (SAGIS), the Hospital Anxiety and Depression Scale (HADS), and the Bristol Stool Scale (BSS) were also used. The adherence to WHO guidelines for aerobic physical activity, and PA levels were measured as Metabolic Equivalent of the Task (MET) minutes per week through the International Physical Activity Questionnaire–Short Form (IPAQ-SF). Microbiota analysis on stool was performed at T0 and T3 in a group of 50 patients. Results: Seventy-five patients with IBS (mean age 41, SD±11.7 years; 70% female) were randomized. Baseline characteristics of two groups were not significantly different. Three patients dropped out in the Not APP group. The improvement in IBS-SSS (88.2±67 vs 47.6±72.0, P=0.021) and in SAGIS total score 9.2±8.7 vs 4.1±8.7, P=0.021) was significantly greater for APP, whereas the difference of the improvement in HADS between groups remained at borderline significance level (4.3±5.0 vs 2.3±3.7, P=0.065). Normalization of stool form was observed in 21% in the Not APP group and in the 41% of patients in the APP group (P= 0.077). Although PA increased in both groups, the increase was greater in the APP than in the Not APP (MET min global activity at IPAQ-SF=791±1026 vs 262±807; P=0.025). The increase of PA was significantly correlated with the improvement of IBS-SSS (r=0.33, P=0.03).Changes in the gut microbiota composition were observed in IBS patients after physical activity. While alpha-diversity remained unchanged, beta-diversity showed significant shifts in microbial community structure from baseline to end of treatment (p=0.001). Specific genera were differentially enriched post-treatment, suggesting possible PA-induced modulation. At the species level, analysis revealed several enriched (amplicon sequence variants) ASVs after intervention.Conclusions: A digital APP promoting PA improved symptoms compared to PA counselling alone. Gut microbiota plays a role in symptom improvement, though it is not the sole driver. APP and increased activity may help shape microbial changes in conjunction with other factors to improve IBS symptoms.
PROMOTION AND MONITORING OF PHYSICAL ACTIVITY WITH A DIGITAL APP IN PATIENTS WITH IRRITABLE BOWEL SYNDROME: CLINICAL OUTCOMES AND FECAL MICROBIOTA ANALYSIS OF A RANDOMIZED CONTROLLED STUDY / A. Costantino ; tutor: M. Vecchi ; coordinator: M. Del Fabbro. Dipartimento di Fisiopatologia Medico-Chirurgica e dei Trapianti, 2025. 37. ciclo
Creative commons - http://creativecommons.org/licenses/by-nc-nd/4.0/
REGULATION AND PHYSIOLOGICAL ROLE OF NON-CLATHRIN ENDOCYTOSIS OF THE EPIDERMAL GROWTH FACTOR RECEPTOR
The epidermal growth factor receptor (EGFR) plays a pivotal role in physiological cellular processes, but its aberrant activation is linked to cancer progression. Endocytosis is a critical regulator of EGFR activity. We have previously identified a novel endocytic route of the EGFR, non-clathrin endocytosis (NCE), which is activated only after stimulation with high doses of the ligand epidermal growth factor (EGF). Unlike canonical clathrin-mediated endocytosis (CME) that directs receptor mainly to recycling, NCE leads mostly to receptor degradation, restricting EGFR signaling and protecting cells from overstimulation. The molecular characterization of the NCE pathway led to the identification of a key functional regulator of NCE: the endoplasmic reticulum (ER)-shaping protein, reticulon-3 (RTN3). This regulator is necessary for the formation of contact sites between the plasma membrane (PM) and the ER during NCE internalization. Localized Ca2+ release at these contact sites induces the fission of NCE vesicles, completing the internalization process. In addition, CD147, a member of the immunoglobulin superfamily, was identified as a cargo co-internalizing with the EGFR in NCE vesicles. CD147 has been validated as a specific marker of NCE, and together with RTN3, represents an invaluable experimental tool for investigating this pathway. In the present work, we have dissected the signaling pathway promoting Ca2+ release from the ER upon EGFR-NCE activation, uncovering a specific role of the phospholipase C G2 (PLCG2) enzyme. PLCG2-dependent Ca2+ release at NCE sites occurs through the activation of the inositol trisphosphate receptor (IP3R) on the ER membrane and is required for the fission of EGFR/CD147-positive NCE vesicles from the PM.At the functional level, we extended the relevance of NCE to alternative EGFR ligands, beyond the EGF. Saturating doses of the two EGFR ligands, amphiregulin (AREG) and transforming growth factor-α (TGF-α), differentially triggered EGFR/CD147 internalization via NCE: while AREG efficiently activated NCE and Ca2+ release at the PM, TGF-α was less effective at triggering this pathway. This finding led us to hypothesize that the differential ability of alternative EGFR ligands to activate NCE could be responsible for the differences in the EGFR fate and biological output exerted by these ligands. Interestingly, stimulation of cells with another growth factor, hepatocyte growth factor (HGF), induced CD147-NCE similarly to stimulation with EGF, implying a broader role of NCE in the regulation of surface proteins. As a model system to test the physiological relevance of NCE, we are using organoids prepared from the non-transformed breast epithelial cell line MCF10A and from primary mouse intestinal crypt and mammary gland epithelial cells. These cells are dependent on EGFR signaling for their growth and differentiation in Matrigel. Our data showed that treatment with compounds that inhibit EGFR-NCE increase growth of mice intestinal and mammary organoids.In conclusion, we have molecularly dissected the signaling pathway leading to EGFR-NCE and expanded its relevance to alternative EGFR ligands and other growth factors. Given its crucial role in downregulating signaling and mediating growth restriction, NCE could behave as a possible tumor suppressor pathway and its regulators could represent novel targets in cancer therapy.
REGULATION AND PHYSIOLOGICAL ROLE OF NON-CLATHRIN ENDOCYTOSIS OF THE EPIDERMAL GROWTH FACTOR RECEPTOR / G. Jendrisek ; added supervisor: S. Sigismund ; internal advisor: G. Scita ; supervisor: P. P. Di Fiore. Dipartimento di Oncologia ed Emato-Oncologia, 2022 Dec 16. 34. ciclo, Anno Accademico 2022.
THE IMPACT OF CLEC4A4 DEFICIENCY ON CARDIOMETABOLIC DISEASES
Introduction and aim Dendritic cell immunoreceptor 2 (DCIR2 or Clec4a4) is a C-type lectin receptor mainly expressed by type 2 dendritic cells involved in T lymphocyte training. Since the strong involvement of the immune system in chronic cardiometabolic diseases, we decided to deep investigate the contribution of Clec4a4 in the development of atherosclerosis and obesity. Materials and methods Clec4a4-/- mice were crossed with Ldlr-/- or ApoE-/- mice to generate double or single KO on atherogenic backgrounds. Male and female littermates were fed for 12 weeks with a Western-Type Diet (WTD) to exacerbate atherosclerosis and histological and flow cytometry analysis were performed. Obesity and metabolic syndrome were induced by feeding Clec4a4-/- and WT male mice with a High-Fat Diet (HFD) for 20 weeks and in vivo glucose and insulin tolerance test and ex vivo histological and flow cytometry analysis were performed. Results Clec4a4 showed a proatherogenic role (-26% and -49% aortic plaque in DKO mice compared to ApoE-/- and Ldlr-/- respectively; p<0.05) with a major effect on circulating neutrophils on male mice (-59% and -47% on ApoE-/- and Ldlr-/- background respectively; p<0.05). Moreover, Clec4a4-/- male mice on Ldlr-/- background revealed an impact on plasma lipid distribution (-59% of cholesterol and -33% of triglycerides; p<0.01, p<0.05 respectively), not observed on ApoE-/- background. In parallel, the data from the obese model showed a comparable immunophenotype between Clec4a4-/- and WT mice after 20 weeks of diet, parallel to a similar hepatic and adipose parenchyma. The exception was represented by the distribution of immune cells within the liver which revealed a significant decrease of dendritic cells (-35%, p<0.05) mainly driven by conventional type 2 DCs impacting on Th2 polarization. Furthermore, Clec4a4-/- mice displayed alterations in the systemic metabolism with glucose intolerance without impacting downstream insulin pathways. ConclusionsOur data showed a redundant role of Clec4a4 in the development of obesity, while a meaningless impact on hepatic dendritic cell distribution and glucose resistance. Our atherosclerosis experimental models showed a proatherogenic role for Clec4a4, with a major impact on circulating neutrophil amount. Future studies will be aimed at understanding the molecular mechanisms existing between this receptor and atherosclerosis.
THE IMPACT OF CLEC4A4 DEFICIENCY ON CARDIOMETABOLIC DISEASES / R. Bellini ; tutor: G. D. Norata ; coordinatore: G. D. Norata. Dipartimento di Scienze Farmacologiche e Biomolecolari, 2023. 36. ciclo, Anno Accademico 2023.
FUNCTIONAL CHARACTERIZATION OF VPS13 DURING SEXUAL AND APOMICTIC PLANT REPRODUCTION: THE MISSING GENE FOR PLANT BREEDING?
In the near future, food production must increase to ensure food security for the growing world population. One of the methods to improve production is the generation of F1 hybrids, which have increased characteristics compared to their parental lines, including higher yield. Apomixis is an asexual way of reproduction that allows the fixation of any increased characteristics for all subsequent generations by the production of a clonal progeny. Despite that, apomixis is absent from economically important crops; therefore, its introduction into sexually reproducing crops could potentially change the plant breeding field. A candidate gene for apomixis regulation in plants is VPS13, identified in apomictic dandelion.During my Ph.D. project, I studied the possible role of VPS13 in plant reproduction in two species, dandelion and Arabidopsis, model plants for asexual and sexual reproduction, respectively. Here, I show that apomictic dandelion maintains the machinery to perform sexual meiosis, repressed by a yet unknown mechanism. Furthermore, the analysis of a knockout mutant suggested that VPS13 might act in concert with other factors to accomplish diplospory in apomictic dandelion.I also report that VPS13S has a crucial role in female germline development in sexual Arabidopsis, demonstrated by the severe defects observed in vps13 mutant ovules. Several analyses suggested the potential function of VPS13S in modulating the expression of key factors at the protein level by regulating their turnover. The results presented in this thesis indicate that the VPS13S association with the membranes is fundamental to its function in ovule development, and they provide insight into the regulatory networks that control ovule development in plants.
FUNCTIONAL CHARACTERIZATION OF VPS13 DURING SEXUAL AND APOMICTIC PLANT REPRODUCTION: THE MISSING GENE FOR PLANT BREEDING? / L. Cornaro ; scientific tutor: L. Colombo ; course coordinator: R. Mantovani. Dipartimento di Bioscienze, 2024 Jun 21. 36. ciclo
A NOVEL, RAS-INDEPENDENT ROLE FOR NF1 IN MICROTUBULE DYNAMICS AND DAMAGE REPAIR DICTATES SENSITIVITY TO T-DM1 IN HER2 POSITIVE BREAST CANCER
Neurofibromatosis 1 (NF1) is among the first tumor suppressor genes to be cloned. Loss-of-function (LoF) mutations in NF1 are at the basis of the onco-developmental syndrome Neurofibromatosis type 1 and are frequently found at the somatic level across multiple tumors. In breast cancer, NF1 LoF has been implicated in resistance to both endocrine and anti-HER2 agents. This has been attributed to the best characterized activity of NF1, its GTPase-activating protein (GAP) function that attenuates RAS signaling.NF1 encodes Neurofibromin (NF1), a large multifunctional protein consisting of 2,818 amino acids. Because of its large size and difficulty in cloning, NF1 structure has resisted characterization for several years and only recently high-resolution reconstructions by Cryo-EM have been obtained. In early studies, NF1 was found to co-purify with tubulin and to co-localize with microtubules (MTs); two regions in NF1 have been proposed to mediate this interaction. The physiological relevance of this interaction remains uncertain, and NF1 is notgenerally included among bona fide Microtubule-Associated Proteins (MAPs).Our understanding of microtubule dynamics has evolved in recent years. It is now recognized that dynamic changes do not occur only at the MT end (“dynamic instability”) but also within the lattice, where the mechanical stress induced by motors or other sources generates discontinuities that are continuously repaired by intra-lattice tubulin incorporation and redistribution of the tip-binding protein EB1. To date, only two MAPs, CLASP and SSNA1 have been found to recognize and modulate such MT damage. None of these proteins is commonly mutated in cancer, so the relevance of MT repair for tumorigenesis remains uncertain, despite the established relevance of MT-dependent functions such as mitosis.Here, combining in vitro/in vivo models and protein biochemistry, we show that NF1 loss sensitizes cancer cells to T-DM1, the first approved Antibody-drug conjugate (ADC) in breast cancer, through a novel, RAS-independent function on microtubular dynamics and repair. NF1 exhibits all biochemical properties of a bona fide MAP and specifically enhances intratubular repair. These results define NF1 as a key regulatory factor for microtubule repair and the first ADC payload-associated predictive biomarker identified to date.
A NOVEL, RAS-INDEPENDENT ROLE FOR NF1 IN MICROTUBULE DYNAMICS AND DAMAGE REPAIR DICTATES SENSITIVITY TO T-DM1 IN HER2 POSITIVE BREAST CANCER / E. Messuti ; tutor: L. Mazzarella co-tutor: S. Minucci. Dipartimento di Oncologia ed Emato-Oncologia, 2025 Oct 01. 37. ciclo, Anno Accademico 2024/2025.
I CONTRATTI DI FIUME NEL SISTEMA DI GESTIONE E TUTELA DELLE ACQUE. NATURA GIURIDICA E FUNZIONI
The river contract has been defined by the legislator as a negotiated planning tool that findsapplication in the complex system of water management and protection. It is a flexible, multilevel,multiobjective, participatory and cross-sectoral tool useful for implementing and defining watermanagement planning and programming at a basin or sub-basin level aimed at the protection, propermanagement of water resources and enhancement of fluvial territories, together with protection fromhydraulic risk, contributing to the local development of these areas.The objective of this thesis is to examine critically whether the river contract can actually be a usefultool for rebalancing the system resulting from the complex regulatory and planning regulations onwater management and protection.To this end, the discipline in the field of water management was preliminarily reconstructed,highlighting the principles that constitute its foundation and focusing attention on the specificinstrument of the river contract, whose historical-legal origin, systematic placement and current stateof implementation in the national experience were reconstructed.Having then identified the critical issues of the water management and protection system withinwhich the river contract is embedded, the study of its legal nature was deepened in order todemonstrate how it does not represent an additional layer in the complex system mentioned above,but a consensual and voluntary instrument capable of assisting the organization between thecompetent administrations, the connection between existing plans and the participation of privateindividuals in the administration of the common good.Further development of the argumentative path required the identification of possible functions of theriver contract capable of responding precisely to the critical issues related to the misalignment ofspatial, functional and temporal scales of the water management system.It was shown, first and foremost, how the river contract can be a useful tool for linking the institutionaland ecological scales. It gathers the requests of local communities, through a participatory process,so that they are considered by superordinate plans, in an ascending process, and, at the same time,regulates detailed actions, in a descending process, constituting an expression of the principle ofvertical subsidiarity.The river contract, then, on a functional level, represents a typical example of multi-objectivemeasures that transcend water quality protection alone, extending also to the containment ofhydrogeological risk and the protection of the neighboring territory and landscape.Furthermore, it was shown how the river contract also possesses a peculiar function of temporallinkage between asynchronous plan instruments and represents an effective tool of adaptivemanagement given the absence of a rigidly fixed duration.Finally, a final function of the river contract has been identified in its ability to represent a useful toolcapable of intercepting a multiplicity of multiobjective funding, thus differentiating itself from thesectoral intervention logics of the past.The question remains open about the future possibility of using the instrument in question not onlyfor the management of the water resource, but of a plurality of natural resources, such as lakes,coastlines, ecological networks and landscapes.
I CONTRATTI DI FIUME NEL SISTEMA DI GESTIONE E TUTELA DELLE ACQUE. NATURA GIURIDICA E FUNZIONI / M. Magnarelli ; tutor: M. Cafagno ; coordinatore: F. Biondi. Dipartimento di Diritto Pubblico Italiano e Sovranazionale, 2023 Jul 05. 35. ciclo, Anno Accademico 2022.
MAKING UP URBAN SPACE. A COMPARATIVE ETHNOGRAPHY OF SOCIAL DISTANCING ON PUBLIC TRANSPORT
While social distancing has emerged as a global strategy in addressing the COVID-19 pandemic, its relevance in social science, particularly as a category for the study of everyday life, remains a subject of debate. This research, relying on the complementary aspects of interactionist studies, neo-materialism, and neo-Marxist perspectives, delves into phenomena and ecologies of social distancing on public transport within Milan, Amsterdam, and New York City. The study, moving beyond the pandemic context, addresses two primary inquiries: how does social distancing emerge from public transport experience? Furthermore, what insights can the relationship between social distancing and public transport provide about urban space as a whole? This ethnographic investigation explains how social distancing produces – and simultaneously is produced by – urban territories through material and symbolic negotiation of geographical, geometrical and imagined distances. This project, approaching the field from different angles, examines social distancing in the choreographies of mobility practices on Milan’s public transit, the configurations of unequal urban experiences in Milan and Amsterdam, and the narratives and collective imaginaries of public transport in Milan and New York. Each case highlights how the ways that people deal with social distancing are situational and mediated by a plurality of heterogeneous actors. This exploration prompts a shift from binary interpretations of material and immaterial realms to a deeper and more comprehensive understanding of urban space.
MAKING UP URBAN SPACE. A COMPARATIVE ETHNOGRAPHY OF SOCIAL DISTANCING ON PUBLIC TRANSPORT / F. La Bruna ; supervisor: P. A. Rebughini ; co-supervisor: G. Semi ; director of doctoral program: P. A. Rebughini. - Milano. Dipartimento di Scienze Sociali e Politiche, 2025 Sep 18. 37. ciclo, Anno Accademico 2024/2025.
THE INFLUENCE OF ANESTHESIA IN THE CENTRAL NERVOUS SYSTEM STUDY
Central nervous system is a complex machine; it is constituted by more or less 172 billions of cells divided between neurons and glia. This incredible number of cells, particularly neurons, are constantly connected, working 24 hours a day, never stopping. Their activity is maintained even during particular conditions such as sleep, pathological unconsciousness (coma) or general anesthesia.For all these peculiarities, since ancient times, man has devoted much effort to the study of the most fascinating organ of living beings (mammals in particular).Nowadays, thanks to the advance in medical technology, many tools are available to “look inside the brain”.Magnetic resonance imaging and, particularly, functional magnetic resonance imaging (fMRI) is a modern biomedical imaging method, which allows a non-invasive assessment of brain function. The detection of brain activity is based on the coupling between neuronal activity, energy consumption, and blood flow.Functional connectivity (FC) of brain regions is modulated in various central nervous system diseases, during sleeping and general anesthesia.Anesthesia during MRI procedures is commonly used in preclinical setting and, sometimes, is required also in clinical setting (uncollaborative patients, children, drugs induced-coma etc).The study of the relationship between anesthetics and FC presents a double value: allows to distinguish the alterations induced by anesthesia on FC, avoiding possible confounding elements, and permits an in depth investigation of drugs behaviour.For all these reasons, the main topic of this PhD dissertation is the relationship between anesthesia and central nervous system. We started from theoretical studies in healthy subjects and we arrived to clinical setting, describing the possible application of anesthetics drugs as a treatment of neurologic diseases.The first study included aims to describe the way in which dexmedetomidine and isoflurane modulate FC in guinea pigs. We analysed the characteristic of cortical, subcortical and cortico- subcortical connectivity under both drugs with resting state fMRI.The second study presented partial results of a more complex work concerning FC in rats under 4 different anesthetics protocols. Because in FC studies blood flow represents a crucial element we dedicated a part of the work to the study of haemodynamic alterations through the administration3of contrast medium. Dynamic Suceptibility Contrast MRI analysis allows the study of cerebral blood flow and cerebral blood volume of different brain anatomic regions under dexmedetomidine, isoflurane, midazolam-dexmedetomidine and midazolam-isoflurane.Finally, we moved to clinical setting to describe the successful treatment of 3 dogs suffering from idiopathic epilepsy presented in emergency department in a state of super refractory status epilepticus. They were treated successfully with a continuous infusion of dexmedetomidine and ketamine.In conclusion, taken together this thesis gives a little contribute to better understand anesthetics behavior at brain level. We believe that it is important to make a “rational choice” when we decide the anesthetic protocol for neuroimaging procedures (both in clinical and preclinical setting) and this “rational choice” could be make only if we have a widespread literature that describe the highest number of anesthetic protocol and their interaction on central nervous system. Finally, thanks to the continuous improvement of our understanding of anesthetics mechanism of action, especially from a molecular and functional point of view, is it possible to use different anesthetics as a therapy for different neurologic conditions, particularly the one based on neurotransmitters imbalance.
THE INFLUENCE OF ANESTHESIA IN THE CENTRAL NERVOUS SYSTEM STUDY / D. Gioeni ; tutor: G. Ravasio ; coordinatore: V. Grieco; curator: M. G. Bruzzone. DIPARTIMENTO DI MEDICINA VETERINARIA, 2020 Feb 06. 32. ciclo, Anno Accademico 2019. [10.13130/gioeni-daniela_phd2020-02-06].
UNA MISTERIOSA DIALETTICA. MOVIMENTO DELLA VITA E SENSO DELL'AZIONE STORICA, A PARTIRE DA HEIDEGGER E PATOČKA
In my doctoral thesis, I intended to start from the following observation: Heidegger's philosophy has shaped the dialectical rift in twentieth-century philosophy in a similar way as Hegel did from the previous century. Taking up the famous formulation used by Marx with regard to the Hegelian dialectic, what I have attempted to do is to separate the «rational core» of Heidegger's dialectic, namely the «general forms of movement» of experience described by it, from its «mystical rind», alluding by the latter term to the «grounds» on which Heidegger found these forms. These grounds, in Heidegger's work, are essentially two: first, the formal concept of «existence», in the phase of his meditation that culminates in Being and Time, and then the «history of being», following the so-called «turn». Similarly to how Marx intended to «make the Hegelian dialectic walk on its feet, rather than on its head», finding its forms not on the mystifying and superstructural level of consciousness and Idea but on the concrete and structural level of the socio-economic organisation of a form of life and the conflicts that run through it, I have considered, starting from the first season of French phenomenology, some of the concrete grounds on which the general forms of movement described by Heidegger have been traced, starting from the plane of corporeality. This «deformalisation» of Heidegger's ontology has served, by contrast, to highlight its extraordinary coherence from the point of view of its formal scheme, which contemporary philosophy has often inherited and maintained to the very extent that it has extended and applied it to theories of experience that intended to demystify both the existential and the historical-ontological planes explored by Heidegger. In my analysis of this formal scheme, I intended, first of all, to highlight its roots, which lie above all in the Heideggerian operation of «ontologizing» the categories of proto-Christian anthropology (Part I); then, its articulation, which defines a movement of «self-extraneation» of experience that differs from that described by the Hegel-Marxian dialectic (Part II). In this way, using an interpretation of Jan Patočka's mature philosophy, I was able to reflect on the limits of the «paradigm» that Heidegger's dialectic has imposed on contemporary philosophy (Part III). Dialoguing mostly with the «ontological-political» reading of the «Heideggerian paradigm» recently proposed by Roberto Esposito, I asked: doesn't the very adherence to the laws of movement described by Heidegger, however much it may be applied to different and more concrete planes than those to which he limited them, have in turn, and in themselves, decisive implications? And what are the risks of these implications, particularly at the level of praxis, of the meaning of action in history, of the posture to be adopted in the face of the question: «what is to be done»? To this question I then connected another: does the application of this form not have, perhaps, limits? Is there not a deeper plane of experience, beyond which it ceases to be valid? And, if so, what relationship does this outlook, this dimension that cannot be colonised by Heideggerian forms of movement, have to the inherent risks of this very form? Is it able to denounce them, mitigate them, dispel them? Or does it not rather risk deepening them?
UNA MISTERIOSA DIALETTICA. MOVIMENTO DELLA VITA E SENSO DELL'AZIONE STORICA, A PARTIRE DA HEIDEGGER E PATOČKA / M.a. Mollisi ; tutor: C. Di Martino; supervisori: T. Campbell, C. Palermo ; coordinatore: N. Guicciardini. Dipartimento di Filosofia Piero Martinetti, 2024 Sep 20. 36. ciclo, Anno Accademico 2022/2023.
A UNIFYING FRAMEWORK TO STUDY THE GENETIC AND ENVIRONMENTAL FACTORS SHAPING HUMAN BRAIN DEVELOPMENT
The development of human brain is a fascinating and complex process that still needs to be uncovered at the molecular resolution. Even though animal studies have revealed a lot of its unfolding, the fine regulation of cellular differentiation trajectories that characterizes humans has become only recently open to experimental tractability, thanks to the development of organoids, human cellular models that are able to recapitulate the spatiotemporal architecture of the brain in a 3D fashion. Here we first benchmarked human brain organoids at the level of transcriptomic and structural architecture of cell composition along several stages of differentiation. Then we harnessed their properties to probe the longitudinal impact of GSK3 on human corticogenesis, a pivotal regulator of both proliferation and polarity, that we revealed having a direct impact on early neurogenesis with a selective role in the regulation of glutamatergic lineages and outer radial glia output. Moreover, we spearheaded the use of organoids for regulatory toxicology through the study of Endocrine disrupting chemicals (EDC), pervasive compounds that can interfere with human hormonal systems. Early life exposure to EDC is associated with human disorders, but the molecular events triggered remain unknown. We developed a novel approach, integrating epidemiological with experimental biology to study the mixtures of EDC that were associated with neurodevelopmental and metabolic adverse effects in the biggest pregnancy cohort profiled so far. Our experiments were carried out on two complementary models i) human fetal primary neural stem cells, and ii) 3-dimensional cortical brain organoids and we identified the genes specifically dysregulated by EDC mixture exposure, unravelling a significant enrichment for autism spectrum disorders causative genes, thereby proposing a convergent paradigm of neurodevelopmental disorders pathophysiology between genetic and environmental factors. Finally, while EDCs are everywhere, their impact on adverse health outcomes can vary substantially among individuals, suggesting that other genetic factors may play a pivotal role for the onset of the disorders. We took advantage of organoids multiplexing to recapitulate, at the same time, neurodevelopmental trajectories on multiple genetic backgrounds, and showed that chimeric organoids preserved the overall morphological organization and transcriptomic signatures of the ones generated from single lines. In conclusion our work shows the possibility to perform population level studies in vitro and use the deep resolution of molecular biology to dissect key aspects of human neurodevelopment.
A UNIFYING FRAMEWORK TO STUDY THE GENETIC AND ENVIRONMENTAL FACTORS SHAPING HUMAN BRAIN DEVELOPMENT / N. Caporale ; internal supervisor: F. Nicassio ; external supervisor: N. Rajewsky ; supervisor: G. Testa. DIPARTIMENTO DI ONCOLOGIA ED EMATO-ONCOLOGIA, 2020 Jan 28. 31. ciclo, Anno Accademico 2019. [10.13130/caporale-nicolo-_phd2020-01-28].
PARTICIPATORY EVALUATION OF AGRICULTURAL DEVELOPMENT PROGRAMS AS A TOOL FOR IMPROVED SUSTAINABILITY IN DEVELOPMENT COOPERATION
‘Every human conduct that is not totally impulsive or just routine mechanics seems to be guided by an evaluation process’ (Dewey, 1939). Switching away from this individual definition of evaluation, the thesis acknowledges the definition given by OECD-DAC and focus the experimentation of well-known multicriteria decision aiding methodologies for ex-ante assessment in a specific development cooperation case study. Chapters 1 and 2, resume the structured bibliographic that has been conducted in order to identify an updated sample of literature about evaluation in development cooperation. During the evolution of the sector, several approaches and methodologies have been conceived and are continuously developing in order to match the variety of evaluanda and contexts. So, three main approaches can be defined, still keeping in mind that they are interlaced in their mutual evolution: the “Positivist-Experimental”, the “Pragmatist-Quality” and the “Constructive-Social Process” approaches group the contributions of several authors, ranging from the more theoretical philosophical basis to the development of specific methodologies and practical techniques. The reality of human development today, as defined by UN through SDGs, requests a multifaceted approach that is capable of mixing the existing approaches, methodologies and techniques. Pluralism of values and focus on the real use of evaluation results through participatory knowledge management are the key elements of the “Constructivist-Social Process approach” which is gaining attention in development cooperation evaluation systems. The literature review took in consideration the state of art of governmental and non-governmental organizations for worldwide and brought to the conclusion that ESs are nowadays in the middle of a renovation process which is far from being concluded. For this reason, we worked in order to provide new insights about sound methodologies able to integrate the different paradigms and methodologies in order to keep methodological robustness and allow a transformative evaluation process to take place in a specific case study. Chapter 3 frames our case study that is located in the Diocese of Goma, Nord Kivu, Democratic Republic of Congo. The framework for this case study is provided by a 3-year EU-funded project called ARDST ‘Appui au retour de réfugiés et déplacés par le biais de la sécurisation de terres en Diocèse de Goma’, led by Caritas Development Goma NGO. The project started in February 2016 and ended in June 2019. In this context our research group was asked to lead a strategic sectoral evaluation. The aim of the evaluation was to inform the choice of interventions meant to trigger sustainable agricultural development in the Diocese of Goma. Chapters 4 to 6 report the pre-print version of three scientific papers issued from the research. Chapter 4 deals with the sketching of a spatial decision support system aimed at Multi-Criteria Evaluation of potential pilot sites for agricultural development and refugees’ resettlement. Land disputes are considered both key sources and perpetuating factors of conflict in the eastern Democratic Republic of the Congo. Existing literature demonstrates that remote sensing is a useful tool for systematically monitor the spatial-temporal land use/land cover dynamics in many regions of the world. For this reason, in this Chapter we propose a methodology for the integration of different sources of information, namely satellite imagery and census information, in order to set up multi-method approach for evidence-based decision-making in development cooperation. Chapter 5 deals with the use of a simplified multicriteria decision-making methodology and namely the application of a simplified form of pairwise ranking (SPWR). International Aid initiatives involve complex real-world decision-making problems in all their relevant phases. Agri-environmental and generally landscape-sc
PARTICIPATORY EVALUATION OF AGRICULTURAL DEVELOPMENT PROGRAMS AS A TOOL FOR IMPROVED SUSTAINABILITY IN DEVELOPMENT COOPERATION / P. De Marinis ; tutor: G. Sali ; coordinatore scuola dottorato: D. Bassi. DIPARTIMENTO DI SCIENZE AGRARIE E AMBIENTALI - PRODUZIONE, TERRITORIO, AGROENERGIA, 2020 Feb 04. 32. ciclo, Anno Accademico 2019. [10.13130/de-marinis-pietro_phd2020-02-04].
INKT CELLS CONTRIBUTE TO COLORECTAL CANCER PROGRESSION INDUCING NEUTROPHILS PRO-TUMORIGENIC FUNCTIONS
Colorectal cancer (CRC) is a multifactorial disease driven by genetic alterations, environmental factors and inflammation. Inflammation is a hallmark of cancer with opposite roles in CRC. Hyperactivation of immune cells can lead to chronic colitis and colitis-associated CRC, equally hyporesponsive immunity can promote disease progression, while anti-tumorigenic immunity efficiently limits tumor growth. Mucosal immunity is involved in the patrolling of intestinal tissues and in the maintenance of homeostasis. Disruption of intestinal homeostasis leads to different pathologies, among which CRC. Invariant Natural Killer T (iNKT) cells are a lipid-specific, CD1d-restricted population of unconventional T cells residing also in the intestinal mucosa. Although their role in inflammation, infection and in tumor immune surveillance has been extensively elucidated, iNKT cells involvement in CRC progression remains controversial.Here, by analyzing a cohort of 118 CRC patients we showed that human tumor-infiltrating iNKT cells acquire a pro-tumorigenic, exhausted phenotype characterized by high expression of PD-1, GM-CSF and IL17, while they maintain cytotoxic properties in the paired non-tumor tissue site. The phenotype of intra-tumor iNKT cells was induced by CRC-associated microbiota and Fusobacterium nucleatum (Fn). Priming with Fn did not influence the killing capabilities of iNKT cells in vitro, while it induced their expression of GM-CSF, IL17 and neutrophil-chemotactic genes. The existence of an iNKT-neutrophil crosstalk was confirmed by ex vivo correlative analysis and in vitro functional assays. Three different murine models of CRC revealed that iNKT cells promote tumor growth inducing a pro-tumorigenic polymorphonuclear-myeloid derived suppressor cell (PMN-MDSC) gene signature in tumor associated neutrophils (TANs). Importantly, in vivo treatment with αGalCer restored tumor control and iNKT cell cytotoxic functions. Survival analyses of human CRC patients showed that co-infiltration by iNKT cells and TANs led to unfavorable prognosis.Our results identified a functional role for iNKT cells in the pathogenesis of CRC, promoting pro-tumorigenic properties on TANs, and suggest a direct implication of the CRC-associated microbiota in the process. The targeted manipulation of iNKT cells restored their anti-tumorigenic properties, thus highlighting their potential use for novel cancer immunotherapies.
INKT CELLS CONTRIBUTE TO COLORECTAL CANCER PROGRESSION INDUCING NEUTROPHILS PRO-TUMORIGENIC FUNCTIONS / G. Lattanzi ; tutor: F. Facciotti ; co-tutor: G. Testa ; internal advisor: S. Marsoni ; internal examiner: M. Pagani ; external examiner: P. Barral. Dipartimento di Oncologia ed Emato-Oncologia, 2023 Apr 13. 34. ciclo, Anno Accademico 2022.
UNA REPUBBLICA SENZA DEMOCRAZIA. PER UNA STORIA TRANSNAZIONALE DEL MODELLO REPUBBLICANO CONSERVATORE TRA FRANCIA, SANTO DOMINGO E STATI UNITI ((1793 - 1804).
The aim of this research project is to investigate and evaluate the transnational nature of the conservative republican model at the end of the 18th century. Moreover, this project examines how this conservative sentiment was conveyed between France, Santo Domingo and the United States through white Santo Domingo exiled settlers’ strategies and political organizations. Their adaptation to the profound changes of the colonial situation, accompanied by their lobbying action between the years 1794 and 1804, propose a new reading of the Atlantic reconfiguraion: originated by the reconquest of their plantations, soon the reflection would be articulated around the concepts of republic and modernity, republic and property, and, consequently, republic and hierarchization of the human beings according to supposed scientific bases.
UNA REPUBBLICA SENZA DEMOCRAZIA. PER UNA STORIA TRANSNAZIONALE DEL MODELLO REPUBBLICANO CONSERVATORE TRA FRANCIA, SANTO DOMINGO E STATI UNITI ((1793 - 1804) / A. Maffei ; tutor: A. De Francesco ; co-tutor: P. Serna. - Milano. Dipartimento di Studi Storici, 2024 Apr 02. 35. ciclo, Anno Accademico 2022.
QUALITATIVE ANALYSIS OF LIPOPROTEINS AND STUDY OF THE METABOLIC PROFILE IN DIFFERENT CLINICAL SETTINGS BY RAMAN SPECTROSCOPY
Raman spectroscopy (RS) is a promising optical technique, able to offer a rapid specific overview of the biochemical composition of the analyzed sample without any preparation or labeling. This non-invasive technique is based on the radiation/matter interaction where a sample is illuminated by a laser which interacts with the vibrational motions of the molecules, leading to the emission of light at wavelengths different from the incident one. This property is used to obtain a spectrum characterized by peaks referable to the biochemical composition of the sample analyzed.This approach has been integrated with conventional biochemical assay to study metabolic disorders such as: obesity (Ob) and type 2 diabetes mellitus (T2DM). RS was used to study the metabolic/biochemical profile of obese diabetic patients undergoing bariatric surgery (BS), to evaluate the impact of the surgery on body composition and systemic metabolism. The results obtained showed that, despite the improvement of the clinical parameters and levels of adipokines, ghrelin and FGF-19 dosed, Raman data identified a persistent alteration of the patients’ biochemical profile in the post-operative follow-up period (6-12 months), underlining the need for more extended longitudinal studies to assess the powerful of RS to monitor the progression of the disease.This method has also been used to study the quality of lipoproteins (LPs), multimolecular complexes responsible for the transport of lipids in the body. Currently, the amount of cholesterol carried by LPs is one of the main parameters used in clinical practice to assess the risk of cardiovascular events. In particular, the level of circulating low-density lipoprotein cholesterol (LDL-C) is considered one of the main risk factors for cardiovascular disease (CVD). It can deposit on the wall of the arteries, causing the formation and progression of atherosclerotic plaques. Conversely, elevated levels of circulating high-density lipoprotein (HDL-C) cholesterol have long been associated with decreased risk of CVD, given their ability to remove cholesterol deposited on arterial walls. However, recent clinical studies have challenged the concept that high HDL-C levels would automatically translate into clinical benefits. In this regard, the research is focusing on the study of the composition and function of the LPs which are altered in different pathological contexts. However, the study of the quality of LPs lacks a universally accepted gold standard. The development of new methods capable of providing indications on the biochemical profile of the LPs, in a rapid and simple way, is therefore of great interest. In this context, LPs extracted from a cohort of obese patients and a control group of healthy subjects were analyzed by RS. The results obtained demonstrated the suitability of RS to obtain an immediate measure of the LPs quality, defined by their biochemical composition and oxidative state.Overall, this thesis work reveals the potential efficacy of the RS in clinical studies and opens the way to a better understanding of metabolic disorders, for an improved definition of CVD risk.
QUALITATIVE ANALYSIS OF LIPOPROTEINS AND STUDY OF THE METABOLIC PROFILE IN DIFFERENT CLINICAL SETTINGS BY RAMAN SPECTROSCOPY / A. Bonizzi ; tutor: F. R. M. Corsi ; co-tutor: S. Mazzucchelli ; coordinator: L. Pinotti. Dipartimento di Scienze Biomediche e Cliniche, 2023 Mar 30. 35. ciclo, Anno Accademico 2022.
This paper deals with arbitration clauses and class action waiver clauses inserted in consumer contract forms. Both clauses are deemed to be valid and enforceable in the Italian law system, although the decisions of the Corte di Cassazione consider the arbitration clause an unfair term, therefore null and void. Arbitration clause, indeed, is not an unfair term since it does not prevent the exercise of the jurisdiction according to article 33, section 2, letter t) of the Italian Consumer Code. Further, arbitration clause does not lead to a material unbalance of the contract according to article 33, section 1 of the Italian Consumer Code: as a matter of fact, arbitration clause does not restrict the rights of action of consumers, as they can recur to a class-wide suit, anyhow. In particular, consumer may recur to class arbitration suits. Similarly, neither class action waiver clause could be considered an unfair term according to article 33 of the Italian Consumer Code, since it does not refrain consumers from recurring to individual suits in order to enforce their rights. Furthermore, class action waiver clause does not lead to a material unbalance of the agreement, since Italian class action system is largely underexploited, yet. Comparative method has been crucial for the scope of work: on one hand, the study of the American model has inspired the work, since Federal Supreme Court already settled arbitration clause issue and class action waiver clause issue by four decisions issued within years 2009/2015. Indeed, the Supreme Court ruled for the validity and enforceability of these clauses, therefore, barring consumers from recurring to class action suits or class arbitration suits as well as to any kind of individual suit, since cost-benefit ratio is totally inefficient. On the other hand, the in-depth analysis of origins of class action in the common law systems revealed the real nature of Italian class action regulations adopted in 2009 and 2019. American class action provision currently in force, i.e. Rule 23 FRCP, is the outcome of principles of representative suit arising from centuries of case law. Indeed, procedural provisions disclaimed by Rule 23 FRCP, especially those regarding the certification phase, are provided in order to grant the plaintiff with a standing to sue in representation of the absent class members: class members which will be actually bound by the decision issued in the class action suit. In other words, procedural prescriptions become the source of the power of the plaintiff to sue in representation of the class. On the contrary, Italian class action regulations do not provide any representative power of the plaintiff, although American rules were the blueprint for the Italian legislator. Therefore, unlike Rule 23 FRCP, Italian class action reg-ulations, instead of being the source of power of the plaintiff to introduce a class action, are just procedural provisions adding to ordinary ones. Thus, the consumer has the right to freely use these special procedural devices and, therefore, consumers are free (i) to renounce to class action suit; or (ii) to adopt class action provisions in a class arbitration regulation.
CLASS ACTION WAIVER CLAUSE / A. Curti ; co-tutor: P. F. G. Giuggioli ; tutor: R. E. Cerchia ; coordinator: M. T. Carinci. DIPARTIMENTO DI DIRITTO PRIVATO E STORIA DEL DIRITTO, 2020 Jan 13. 32. ciclo, Anno Accademico 2019. [10.13130/curti-adriano_phd2020-01-13].
STUDY OF PHYSIO-PATHOLOGICAL MECHANISMS AT THE BASE OF SCHWANNOMA DEVELOPMENT
Schwannomas are the most common type of peripheral nerve tumor. Usually, schwannomas are benign, well‐circumscribed tumors clinging to peripheral nerves, consisting of a clonal population of Schwann cells (SCs), which often undergo cystic and degenerative changes. Most cases are sporadic, however, some are associated with neurofibromatosis type 2 (NF2), schwannomatosis (SCH) or the pathology called Carney's complex. The schwannoma affecting the auditory nerve, called vestibular schwannoma (VS), is the most common benign tumor of the cerebellopontine angle. The occurrence of VSs is frequent among patients with NF2, the autosomal dominant multiple syndromes resulting from mutations in the NF2 gene, coding for the protein merlin. The tumor suppressor merlin is a cytoskeleton-associated protein, which loss of function is the main cause for transformation of SCs into schwannomas. However, a large amount of VSs is sporadic and unilateral suggesting the presence of unknown pathogenic mechanisms beside NF2 gene mutation. Also, SCH is due to changes in NF2 gene, although recently other mutations in SMARCB1 and LZTR1 genes were found. Considerable overlap has been found between SCH and NF2, in terms of the occurrence of associated types of tumors, whereas both diseases are considered as separated clinical entities. Therefore, it is not excluded that mutations in other genes, may be responsible for the appearance of schwannomas presenting a characteristic clinical phenotype. The identification of such genes and/or cellular mechanisms has found limitations in the lack of in vitro cell models of human origin that may be useful to this purpose. In this thesis, human SCs were isolated, purified and characterized, for the first time, from the schwannomas, by using a positive immunomagnetic cell labelling technique, allowing the specific and high-purity enrichment of SCs in culture. The SCs from peripheral schwannoma were then immortalized using the LtAg-SV40, in order to obtain a continuous cell line for future studies addressed to better understand the schwannoma pathogenesis.Notably, the onset of schwannoma may be influenced by the exposure to environmental challenges, likely the electromagnetic fields (EMF), such as those generated by mobile phones or electronic devices. In fact, it was suggested that EMFs can induce cellular changes towards non-physiological behavior, that may be pathologically relevant to the development of schwannoma. Overall, in this work of thesis we focused our attention on the investigation of putative genes, pathways, and molecular targets responsible for the oncotrasformation of SCs, studying the possible factors, also environmental, that could induce the development of schwannomas. The data are promising but further confirmatory studies will be needed for a more direct cause–effect correlation between EMF and SCs oncotransformation and finally to arrive at definitive conclusions widely accepted by the international scientific community.
STUDY OF PHYSIO-PATHOLOGICAL MECHANISMS AT THE BASE OF SCHWANNOMA DEVELOPMENT / V. Melfi ; tutor: V. Magnaghi; coordinatore: G.D. Norata. Dipartimento di Scienze Farmacologiche e Biomolecolari, 2022 Dec 12. 35. ciclo, Anno Accademico 2022.
LO SPECCHIO DI SÉ. 'AMBASCIATORI', GESUITI E SOVRANI IN SCENA NEL VIAGGIO ITALIANO DI QUATTRO PRINCIPI GIAPPONESI (1585)
"This thesis investigated how the image of Japan was exploited and adapted to the needs of 16th century Europe during the Counter-Reformation.Through the mediation of the Society of Jesus, active in the Archipelago since 1549, Japan has taken on well-defined traits in the European imaginary. On the one hand, those of a culture with which dialogue took place with simplicity on the basis of a - supposed - religious similarity, and on the other, that of a still ""wild"" world that was to be tamed through the religious and cultural colonization carried out by the Fathers.This idea became predominant during the 1580s, when the Jesuits conducted an embassy of four young princes to the main Italian courts to make their narration immediately identifiable: the idea of Japan and its representation would have to match perfectly.The journey, which lasted from 1582 to 1587, with the return to Japan only in 1590 due to the changed political conditions of the Archipelago, was the first moment in which the image of the ""other"", until then confined to the imaginary, found its reification. In a ceremonial journey in which scriptwriters, actors and spectators each played their own role, the Peninsula found itself to be the stage covered by the young princes with their double role: symbol of missionary successes and at the same time medium through which the sovereigns challenged each other with a political exploitation of welcome.The delegation can also be used to analyse the Italian society at the end of the 16th century, with a privileged focus on the mass of spectators. The citizens, in fact, had to be gratified and educated through the theatrical representation of the ""different"" who had to be primarily conquered from a cultural point of view."
LO SPECCHIO DI SÉ. 'AMBASCIATORI', GESUITI E SOVRANI IN SCENA NEL VIAGGIO ITALIANO DI QUATTRO PRINCIPI GIAPPONESI (1585) / A. Tripepi ; tutor: G. Civale, M. M. Benzoni ; coordinatore del dottorato: D. Saresella. DIPARTIMENTO DI STUDI STORICI, 2020 Feb 05. 32. ciclo, Anno Accademico 2019. [10.13130/tripepi-alessandro_phd2020-02-05].
NATURAL AND NATURE-INSPIRED COMPOUNDS AS ANTIMICROBIAL AGENTS: EXTRACTION, SYNTHESIS AND BIOLOGICAL INVESTIGATION
Nowadays the presence of agents and pathogens in food supply chain that developed antimicrobial resistance (AMR) is an increasing problem for human health. World Health Organization (WHO) defined the antimicrobial resistance as one of the top five public health threats. Antimicrobial resistance can be transmitted from microorganisms to microorganisms in different ways (phages, trasposoms and plasmins), resulting in horizontal gene transfer (HGT). It is important to highlight that antimicrobials are used in all sectors of food supply chain: as antifungal and pesticides to maximise the crops; as antibiotics in animal production and in aquaculture; as preservatives for the shelves food life; as antiviral agents in functional foods. The escalating emergence of AMR in food supply chain make it necessary the search for new antimicrobial compounds. In this context, nature can be a potent source of inspiration for the development of new and innovative antimicrobials.The focus of this PhD thesis was to contribute to the expansion of the arsenal of antifungal, antibacterial and antiviral compounds by paying particular attention to natural and nature-inspired compounds. Two different strategies were followed.The first strategy was based on the development of a synthetic approach to build novel antifungal and antibacterial agents. In the field of antifungals, we concentrated on the design and synthesis of a collection of strobilurin derivatives. Strobilurins, or more correctly quinone outside inhibitors (QoI), originate from a natural compound discovered in 1977 and cover almost 20% of the fungicide’s world sales. In particular, this class of compounds is used against Pyricularia oryzae, a rice blast responsible of the biggest part of losses of global production. In the last decades, several resistance mechanisms, based on the mutation in the target site gene of strobilurins, have been developed. Mutation G143A is by far the most important mechanism of resistance and has a strong effect on the activity of QoIs. For this reason, our strategy was based on the development of multi-target compounds, which can improve the efficacy and overcame the resistance, acting simultaneously on multiple, well-known and validated targets. In fact, starting from strobilurins’ pharmacophore, we created new analogues connecting in different position the pharmacophore of another class of antifungal agents, e.g. succinate-dehydrogenase inhibitors. The novel compounds were tested against wild-type and resistant-strobilurin strains of P. oryzae. In silico modeling studies gave important information about the structural requirement of the new antifungal compounds. About antibacterial and preservatives agents, we focused on stilbenoids. Stilbenoids belong to the class of polyphenols, in which resveratrol is the most studied compound. They are produced by plants as means of defence against pathogens invasion and stress factors. Several studies showed their surprising biological activities, such as anticancer, antioxidant, neuroprotective and antimicrobial. In particular, we investigated dimers of resveratrol such as dehydro--viniferin and dehydro--viniferin, carrying out structure-activity-relationship (SAR) studies. Moreover, we modified the scaffold of monomeric stilbenoids with the aim of obtaining compounds able to overcome the outer membrane of Gram-negative bacteria.The second approach was directed to extraction and isolation of bioactive natural products from vegetal matrices. In order to give our contribution on the research against COVID-19, we focused on grapefruit seeds, inspired by the known antiviral activities of grapefruit seeds extract (GSE). A small collection of extracts and five pure isolated secondary metabolites, belonging to the class of limonoids and flavonoid glycosides, were isolated and investigated for the ability to counteract SARS-CoV-2 infection, by studying both the potential
NATURAL AND NATURE-INSPIRED COMPOUNDS AS ANTIMICROBIAL AGENTS: EXTRACTION, SYNTHESIS AND BIOLOGICAL INVESTIGATION / G. Catinella ; tutor: S. Dallavalle; coordinatore: A. Pagliarini. Dipartimento di Scienze per gli Alimenti, la Nutrizione e l'Ambiente, 2021 Dec 03. 34. ciclo, Anno Accademico 2021.
LIPID COMPOSITION OF MICROGLIAL CELLS AND EFFECTS OF A REMYELINATION-PROMOTING ANTIBODY ON THE SPHINGOLIPID PATTERNS
Multiple sclerosis (MS) is one of the most prevalent demyelinating diseases among young adults (Lemus et al., 2018). Myelin damage and oligodendrocyte death are caused by the autoimmune attack of T cells that cross the blood-brain barrier resulting in the formation of sclerotic plaques (Pearce, 2005, Lubetzki and Stankoff, 2014). In the early stages of the disease, damaged myelin is to some extent repaired, however the process is inefficient due to several factors, including impairment in OPCs recruitment and/or OLs differentiation likely due to the inflamed environment, and the inhibitory effect of myelin debris present at the lesion sites (Barres, 2008, Lassmann, 2014, Domingues et al., 2016). There are currently no cures for multiple sclerosis, only treatments that aim at alleviating its symptoms or slowing down its progression. Therefore, interventions that stimulate and promote endogenous myelin repair mechanisms may be a feasible treatment option (Lemus et al., 2018). In animal models of demyelination, the ability of certain monoclonal antibodies to stimulate myelin sheath synthesis has been proven. In this regard, the rHIgM22 monoclonal antibody has showed particularly promising results. This antibody seems to act mainly on cells of the oligodendrocyte lineage, in particular on oligodendrocyte precursor cells promoting their proliferation and inhibiting their apoptosis (Warrington et al., 2000). However, experimental evidence suggests that not only OPCs and OLs are targeted by rHIgM22, but also other glial cells, e.g., astrocytes and microglia, In particular, it has been shown that rHIgM22 stimulates an increase in the phagocytic activity in microglia (Zorina et al., 2018). The phagocytosis of myelin debris by microglia is one of the most delicate and limiting steps in the myelin repair mechanism (Neumann et al., 2009, Domingues et al., 2016, Napoli and Neumann, 2009, Lloyd and Miron, 2019).The protective effects exerted by rHIgM22, at least towards the primary targeted glial cells, appear to be mediated by the reorganisation of membrane lipids leading to the recruitment of a multimolecular complex that results in the development of numerous signalling mechanisms (Watzlawik et al., 2010, Watzlawik et al., 2013a). Indeed, data obtained in our laboratory showed that different glycosphingolipids were involved in rHIgM22 binding at the cell surface but also that rHIgM22 treatment induced changes in the sphingolipid pattern of oligodendroglial cells (Grassi et al., 2015; S. Grassi et al., 2021). Since microglia seem to be a crucial player in the processes that are at the center of myelin repair, they have been the focus of this thesis. We used as an experimental model the BV-2 microglial cell line. The BV-2 cell line is widely used because it retains the morphological, phenotypical, and functional characteristics described for freshly isolated primary microglia and has functional markers of macrophages (Blasi et al., 1990). BV-2 is a semi adherent cell line, where adherent and floating cells may represent two different sub-populations. Due to the paucity of information in the literature regarding the lipid composition of microglia, as the first step, we analysed the lipid composition of BV-2 cells using a combination of traditional chemical analysis and lipidomic using ESI-mass spectrometry. We analysed lipids in different culture conditions since it is well known that culture conditions affect the composition of certain lipid classes (Frechin et al., 2015). We identified differences in the lipid pattern depending on cell culture conditions but also some differences in the lipid composition among adherent and floating cells. Specifically, we observed key differences between adherent and floating cells at 80–90% confluence. In this experimental condition, ceramide was present at higher levels in the floating cells while sphingomyelin was more enriched in the adherent
LIPID COMPOSITION OF MICROGLIAL CELLS AND EFFECTS OF A REMYELINATION-PROMOTING ANTIBODY ON THE SPHINGOLIPID PATTERNS / C. D'aprile ; tutor: A. PRINETTI ; coordinatore: C. SFORZA. Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, 2023 Apr 18. 35. ciclo, Anno Accademico 2022.
ANALYSIS OF WHOLE-BLOOD TRANSCRIPTOME DIFFERENCES AND FUNCTIONAL INFERENCES IN SUBJECTS WITH NON-OBSTRUCTIVE OR OBSTRUCTIVE CAD, DETECTED BY CCTA, COMPARED TO SUBJECTS WITHOUT CORONARY ATHEROSCLEROSIS
Background - Despite the presence of the same CAD symptoms and pre-tests suggestive for the presence of CAD, a not irrelevant number of coronary computed tomography angiography do not detect the presence of coronary atherosclerosis. In fact, individuals could share the same symptoms even having different disease profiles from a biological point of view. Thus, there is an urgent unmet need of new tools/gene-expression patterns that allowed to detect the presence and the severity of coronary artery disease. In this context, the aim of this thesis project is to identify, through RNA-sequencing: i) gene-expression patterns in the whole blood that could differentiate patients with obstructive and non-obstructive coronary atherosclerosis, detected by coronary CT angiography (CCTA), from subjects without the presence of coronary stenosis; ii) biological processes and/or pathways associated to obstructive or non-obstructive CAD.Methods - A total of 730 consecutive patients, undergoing coronary computed tomography angiography (CCTA) for suspected CAD, have been enrolled and coronary stenosis was quantified. Total RNA was isolated from whole blood collected at baseline. The whole-blood transcriptome of 315 patients, matched for age, sex and with (obCAD, n=105) or without (non-obCAD, n=105) obstructive coronary stenosis and of subjects with complete absence of coronary atherosclerosis (noCAD=105), was profiled and compared by RNA-sequencing. Statistical and data analysis was performed with R packages, GSEA and GraphPad Prism.Results - Differential gene expression analysis reveals that whole blood gene expression patterns could differentiate obstructive and non-obstructive CAD patients from subjects with the complete absence of coronary atherosclerosis. Gene-Set Enrichment Analysis revealed that both obstructive and non-obstructive CAD were negatively associated to the regulation of innate and adaptive immunity. Non-obstructive coronary artery disease were positively associated to B cell receptor signalling pathway while obstructive CAD to circadian rhythms.Conclusions - Whole-blood transcriptome profiling revealed differences between patients with obstructive or non-obstructive coronary artery disease as detected by CCTA and subjects with complete absence of coronary atherosclerosis. The main findings confirm the hypothesis that the integration of non-invasive imaging and whole-blood transcriptional profiles could be useful for new tools/blood-based test, able to detect the presence and the severity of coronary atherosclerosis as detected by CCTA and find biological processes associated with obstructive and non-obstructive CAD.
ANALYSIS OF WHOLE-BLOOD TRANSCRIPTOME DIFFERENCES AND FUNCTIONAL INFERENCES IN SUBJECTS WITH NON-OBSTRUCTIVE OR OBSTRUCTIVE CAD, DETECTED BY CCTA, COMPARED TO SUBJECTS WITHOUT CORONARY ATHEROSCLEROSIS / C. Vavassori ; supervisore: G. I. Colombo ; tutor: P. Agostoni ; coordinatore: C. Sforza. Dipartimento di Scienze Cliniche e di Comunità, 2023 Apr 20. 35. ciclo, Anno Accademico 2022.
SELECTIVE ASSEMBLY, PHASE TRANSITIONS AND MOLECULAR KINETICS OF DNA OLIGOMERS.
In the last few years it has been shown that the spontaneous self-assembly process of short DNA and RNA duplexes into liquid crystal ordering is a likely potential route that led to the formation of first nucleic acids able to support biological activities. In particular, it has been experimentally demonstrated that liquid crystal domains behave as suitable micro-reactors to trigger polymerization between the stacked and not initially chemically linked short nucleic acids. Even paired mononucleotides at high enough concentration exhibit liquid crystal ordering, unveiling the crucial role of Watson-Crick selectivity and stacking attractive interactions among base pairs. In such a possible prebiotic context, DNA sequences with both random nucleobases sequence and length are likely to be formed. Surprisingly, it has been shown that even random DNA sequence of fixed length can support liquid crystal ordering at high concentration. The aim of this PhD thesis is to extend the knowledge of DNA liquid crystals self-assembly in the following four directions.First, I explored the selectivity of interaction in nucleic acids solutions of random-sequence DNA oligomers of different length L. The combination of experimental results and a suitable developed theoretical model revealed a not negligible percentage of perfect duplexes.Second, I investigated the process that leads to the onset of the nematic liquid crystal phase in aqueous solutions of DNA duplexes. The combination of static light scattering experiments and computer simulations made possible the study of both aggregation and local ordering of DNA duplexes in the isotropic phase, where no positional order is developed, and in proximity of the isotropic-nematic phase boundary. This study gives an insight of the role on the development of local orientational order among DNA duplexes both far and in proximity of the isotropic-nematic phase boundary.Third, I studied the diffusion of short DNA duplexes with attractive and repulsive interactions in the isotropic phase as a function of temperature. I found that the temperature dependence of diffusion coefficients reflects via an Arrhenius law the interduplex attractive interactions, whereas diffusion of repulsive duplexes is partially well described in terms of repulsive hard spheres.Fourth, I investigated phase diagrams of mixtures of DNA single strands and duplexes with various polycations that show liquid-liquid phase separations. This phenomena leads to the onset of a concentrated but still liquid phase of polyelectrolytes, called coacervate, in a bulk phase where polyelectrolytes are diluted. The most surprising result I found, it is the insurgence of liquid crystals in coacervates with 12 nucleobases long random DNA oligomers and polylysine at different ionic strengths.I believe that this PhD thesis adds important pieces to the self-assembly of nucleic acids puzzle, and in particular it shows how randomness of nucleic acids is not an impasse to both hybridization of defectless duplexes and liquid crystal ordering.
SELECTIVE ASSEMBLY, PHASE TRANSITIONS AND MOLECULAR KINETICS OF DNA OLIGOMERS / S. Di Leo ; supervisor: T. Bellini ; co-supervisor: T. Fraccia. Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, 2022 May 10. 34. ciclo, Anno Accademico 2021.
Parton Distribution Functions (PDF) are the theoretical object describing the colliding hadron structure, arising from the factorization property of perturbative Quantum Chromodynamics (QCD).However, these objects encode the content of the non-perturbative dynamics of large scale QCD, and it is not currently possible to compute them from first principles with the available techniques. But their universality, that is a direct consequence of factorization, allows determining PDFs from experimental data, and use the extracted value to make predictions even for any kind of processes.The main part of this thesis is dedicated to provide fast and reliable theory predictions to be used in a global PDF determination, developing a set of essential tools to make this process scalable and reproducible.Furthermore, some specific early applications of the tool set are presented, leading to remarkable results regarding proton properties.A final part is dedicated to some methodological improvements in the PDFs extraction techniques, following a novel theoretical analysis, that lead to new insights, or taking into account an alternative statisticalframework.
THEORY PREDICTIONS FOR PDF FITTING / A. Candido ; tutor: S. Forte. Dipartimento di Fisica Aldo Pontremoli, 2023 Jan 26. 35. ciclo, Anno Accademico 2022.
DIRITTI FONDAMENTALI E PREVENZIONE DEL TERRORISMO NEL SISTEMA MULTILIVELLO. ALLA RICERCA DI UN BILANCIAMENTO TRA ESIGENZE DI SICUREZZA E TUTELA DELLE LIBERTÀ AI MARGINI DELLA 'MATERIA PENALE'
"The thesis focuses on some of the terrorism prevention tools in the light of the related guarantee statute; in particular, the listing systems developed by the UN and the European Union were considered, as well as the prevention measures governed by Legislative Decree 6 September 2011, no. 159; the attempt is to identify the common thread that unites these apparently very distant measures. From this point of view, the work moves along a twofold direction: on the one hand, the identification of the guarantees that must guard the prevention institutes, also in light of their legal nature in the perspective of the conventional concept of ""criminal matter""; on the other hand, the reconstruction of the discipline of the institutes, based on national and supranational sources and jurisprudence.This analysis is contained mainly in Chapters III and IV, dedicated respectively to the identification of a possible guarantee statute valid for the prevention system as a whole and the reconstruction of the positive discipline of the institutions considered. This part, which constitutes the main focus of the work, is preceded by two chapters, with function of theoretical (Ch.. I) and historical framework (Ch. II).The first Chapter, in fact, wants to provide a frame of the conceptual coordinates in which we move in the next part of the work; it is ideally divided into three parts, which represent the three pillars of the entire thesis. The first axis, that of guarantees, is the object of attention by reconstructing the conventional and constitutional concept of ""criminal matter""; the second, in some ways opposed to the previous one, focuses on the analysis of the concept of ""security"", understood as the value that the entire preventive anti-terrorism system is aimed at protecting. Finally, the last axis represents an attempt to synthesize the two areas already outlined, through the so-called balancing mechanism in terms of proportionality.The scheme adopted in the first Chapter is therefore re-proposed in the following and central part of the work, which has already been mentioned: the issue of guarantees is taken up and elaborated on in Chap. III; in Chap. IV, then, the analysis focuses on the institutes elaborated by the national and supranational legislator with the aim of guaranteeing “security”; in the fifth and final Chapter, an attempt is made to sift - with a view to balancing - the mechanisms set up to protect security claims in the light of fundamental individual guarantees. It is here that attempts have also been made to put forward some proposals for remodeling the system, in order to eliminate some of the aspects of its incompatibility with fundamental rights.The issue of the legal nature of the institutes examined, in the perspective in which it has been placed, is therefore played down by the centrality attributed to the criterion of proportionality as a balancing tool between opposing needs. Similarly, categories such as ""enemy criminal law"", ""struggle criminal law "" and ""emergency law"", even if considered in the first Chapter, are not used critically. The chosen perspective, in fact, is not that of an all-encompassing evaluation in terms of legitimacy-illegitimacy of the entire system, but, rather, an analysis that is as precise and specific as possible of the institutions examined in the light of the guarantee statute. elaborated by the Courts of Rights."
DIRITTI FONDAMENTALI E PREVENZIONE DEL TERRORISMO NEL SISTEMA MULTILIVELLO. ALLA RICERCA DI UN BILANCIAMENTO TRA ESIGENZE DI SICUREZZA E TUTELA DELLE LIBERTÀ AI MARGINI DELLA 'MATERIA PENALE' / P. Bernardoni ; tutor: C. Meloni ; coordinatore: C. Luzzati. Dipartimento di Scienze Giuridiche Cesare Beccaria, 2021 Jun 24. 33. ciclo, Anno Accademico 2020. [10.13130/bernardoni-pietro_phd2021-06-24].
IMPACT OF VITAMIN D DEFICIENCY, DYSLIPIDEMIA AND OBESITY ON SERUM LIPIDOMIC PROFILE. SEARCH FOR NEW BIOMARKERS AS EARLY PREDICTORS OF OBESITY-ASSOCIATED COMORBIDITIES.
Research studies indicate that up to 58% of the world adult population will be overweight or obese by 2030 [1]. Obesity is not only related to food intake, but factors such as lifestyle and genetic background contribute to its onset [3–5]. This disease [2] is commonly associated with vitamin D (Vit. D) deficiency [6,7], and genetic associations have been identified to explain this link [8]; however, differences in dietary intake, sun exposure, or Vit. D metabolism are also involved [9]. Moreover, along with Vit. D deficiency, lipids, and particularly sphingolipids (SLs) as ceramides (Cers) and sphingomyelins (SMs) have been described as involved, not only in increasing inflammation [10,11], but also in the development of cardiovascular disease and type two diabetes [12–14], two common conditions observed in obese subjects. Given the pivotal role of SLs in obesity associated co-morbidities and the association of Vit. D, dyslipidemia and obesity, our study was aimed at profiling circulating SLs in human subjects under these conditions, in order to provide hints for the identification of new biomarkers to be introduced in clinical settings.To define human SLs profile in obesity, dyslipidemia, and Vit. D associated deficiency, sera from 23 normal-weight normolipidemic (NWNL), 46 normal-weight, dyslipidemic, Vit. D deficient (NWDL) and 60 obese dyslipidemic, Vit. D deficient (ODL) Saudi Arabian subjects were analyzed with a dual approach, characterized by the use of two complementary techniques: the HPTLC-Primuline profiling and the LC-MS analysis. Furthermore, to define SLs profiles in the context of human adaptation to high altitude hypoxia, sera from 59 Vit. D deficient dyslipidemic children living at high altitude were analyzed by LC-MS. Children were grouped based on their BMI percentiles in 7 underweight (UW), 30 normal-weight (NW), 13 overweight (OW) and 9 obese (O).SLs profile analysis of NWNL and ODL Saudi Arabian subjects displayed differences in total Cer and total SM caused by dyslipidemia and vitamin D deficiency, whereas specific Cers, and SMs acyl chains characterize obese subjects, only. Gender differences were found in SLs profiles independently from dyslipidemia and Vit. D status. Obesity-associated Cers, SMs, and dihydrosphingomyelins (dhSMs) specific acyl chains were identified in the NWDL vs. ODL comparison independently from dyslipidemia and Vit D status, and are thought to be drivers of increased risk of developing obesity-associated morbidities.The analysis of SLs profiles from dyslipidemic children with Vit. D deficiency allowed to confirm the results of Saudi Arabian subjects regarding SLs association with dyslipidemia and associated Vit. D deficiency. Furthermore, SLs profile analysis led to the identification of a characteristic SLs portraits associated with BMI and related to hypoxia metabolic adaptation.
IMPACT OF VITAMIN D DEFICIENCY, DYSLIPIDEMIA AND OBESITY ON SERUM LIPIDOMIC PROFILE. SEARCH FOR NEW BIOMARKERS AS EARLY PREDICTORS OF OBESITY-ASSOCIATED COMORBIDITIES / P. Barbacini ; tutore: C. Gelfi ; supervisore: J. Casas, V. Hirschler ; coordinatore: M. Samaja. DIPARTIMENTO DI SCIENZE BIOMEDICHE PER LA SALUTE, 2020 Jan 29. 32. ciclo, Anno Accademico 2019. [10.13130/barbacini-pietro_phd2020-01-29].
Women account for about half of the world’s population, and they generally faceseveral barriers to participation in the labor force, run their business, and reach thehigher echelon of their careers. Culture, religious views, and gender-related stereotypesare among the major identified constraints (Comi et al., 2019). A burgeoningliterature in economics seeks to understand the effect of top managers’ or boardof directors’ gender on firm performance. However, prior studies have focused onthe cases of developed countries such as Italy, Norway, among others. (Comi et al.,2019; Ferreira, 2009; Flabbi et al., 2016; Kenneth and Amy, 2012).This thesis attempts to contribute to the research gap by examining the relationshipbetween the gender of the owners or the board directors and the firm performancein two developing countries—Brazil and Malaysia. In the first chapter, I specificallyfocus on the role of gender in firm performance and access to credit in the case ofBrazil. In the second chapter, instead, I explore the impact of gender diversificationof the board of directors on firm performance in the case of Malaysia. In contrast tosome studies that focus on the difference in firm performance based on the gender ofowners or board directors, this study investigates multiple dimensions of firm performance.This thesis uses various firm performance metrics based on the availableinformation in each dataset. The number of employees, total revenue, sales growth,employee growth, labor productivity, value-added per worker, and export propensityis considered for the study in case of Brazil; while total revenue, asset, profitmargin, Return on Equity (ROE), and Tobin’s Q are used for the study in case ofMalaysia. This thesis, therefore, provides a well-rounded and compressive analysisof the role of gender of owners or board of directors in firm performance. Further,it addresses the impact of gender diversification among the board of directors onfirm performance. The majority of prior studies use cross-sectional data, makingit difficult to identify causal links (Hillman et al., 2007; Julizaerma and Mohamad,2012; Post and Byron, 2014). It also considers the influence of other factors such asage, education, and experience of board directors and investigates the existence ofheterogeneous effect of gender diversification among the board of directors based onthe firm size, sectors, and the status of competitiveness of the firm. Furthermore,it explores an important point that never explored in connection with gender diversification,which is the relationship between the share of directors having familyrelationships and firm performance. It also investigates the impact of gender diversificationrequirement in the Malaysian board of directors on the share of directorswho have family relations.
ESSAY ON GENDER AND FIRM PERFORMANCE / B.g. Firew ; PhD program Director Eduardo Rossi; supervisors: T. Frattini, A. Maida. Dipartimento di Economia, Management e Metodi Quantitativi, 2020 Jul 24. 32. ciclo, Anno Accademico 2019. [10.13130/firew-banchayehu-girma_phd2020-07-24].
OPENING NEW CHEMICAL SPACE IN DRUG DISCOVERY
Small molecules have always been of interest in chemistry and biology because of their ability to exert powerful effects on the functions of macromolecules comprising living systems. Screening collections or libraries of small molecules is the most common way to identify new chemical modulators of protein function and constitutes one of the bases of medicinal chemistry. While a general consensus has emerged that library size is not decisive, the concept of library diversity, in terms of molecular structure and, more importantly, molecular function, has become crucial, making the efficient creation of functionally diverse small molecule collections a formidable challenge. When a specific biological target is chosen, the compounds considered in the screening process are usually selected or designed on the basis of knowledge of the target structure or mimicking the structure of known natural ligands. Their synthesis can be planned effectively with retrosynthetic analysis (Target-Oriented Synthesis, TOS).However, when the precise nature of the biological target is unknown, the structural features required in potentially interacting small molecules cannot be defined a priori, and a productive approach can be the synthesis and evaluation of molecular libraries that achieve wide coverage of chemical space (Diversity-Oriented Synthesis, DOS). Indeed, the overall aim of DOS is to generate collections of small molecules with a high degree of structural, and thus functional, diversity that interrogates large areas of biologically-intriguing chemical space simultaneously. Within these two general synthetic strategies, the research herein described is focused on the development of new molecular scaffolds, which, in some cases, proved to be promising lead compounds. Within different synthetic approaches, the high potential of the multicomponent reactions with unprecedented components has been exploited and, on the other hand, new protocols have been finely tuned to achieve new transformations. The synthesis of promising bioactive compounds and the creation of chemical libraries, mainly focused on boron-containing derivatives and oxindole-based molecular frameworks, have been successfully carried out.
OPENING NEW CHEMICAL SPACE IN DRUG DISCOVERY / M. Manenti ; tutor: A. Silvani ; coordinatore D. Roberto. Dipartimento di Chimica, 2024 Jan 29. 36. ciclo, Anno Accademico 2023.
NUTRITION AND FRAILTY: HOW TO CAPTURE THE COMPLEXITY OF OLDER PEOPLE
The older population is characterized by a high clinical complexity. Poor nutritional status is highlyprevalent in older people and has been repeatedly evoked as a major contributor to several health-related negative outcomes. The aim of this thesis was to evaluate the contribution of nutrition in someage-related conditions, taking into account the older people’s complexity.Study I. We explored the association between lack of energy and malnutrition in nursing homeresidents. Lack of energy was highly prevalent in our sample (i.e., 43.2%). We found a significantassociation between malnutrition and lack of energy, independent of potential confounders. Decreasein food intake, reduced mobility, and psychological stress or acute disease in the past 3 months wereidentified as the items of the Mini Nutritional Assessment contributing the most to the associationbetween malnutrition and lack of energy.Study II. We investigated the association between unintentional weight loss and mortality in apopulation of older patients undergoing hemodialysis. Older patients experiencing unintentionalweight loss during the follow-up period had a 3.4-fold risk of death independently of comorbiditiesand other risk factors.Study III. We explored the prevalence of frailty and its association with hospitalization and mortalityin a population of older patients undergoing haemodialysis. Study III outlined a high prevalence offrailty (i.e., 55.2%) among older patients undergoing haemodialysis. Frailty was significantlyassociated with a higher risk of hospitalization and all-cause mortality in our sample.Conclusions. Nutritional status is an aspect frequently overlooked, despite it has been repeatedlyevoked as a major determinant of health status in all stages of life. Our findings clearly show the roleof nutritional status as a mediator of several health-related conditions in older people. Indeed, giventhe burden it poses in terms of adverse health outcomes, increased (research) efforts should be paidto nutritional status. A multidimensional and multidisciplinary approach is pivotal for the early identification of people presenting nutritional issues in order to prevent the onset of seriouscomplications.
NUTRITION AND FRAILTY: HOW TO CAPTURE THE COMPLEXITY OF OLDER PEOPLE / D. Azzolino ; tutor: M. Cesari; coordinatore: L. Pinotti. Dipartimento di Scienze Cliniche e di Comunità, 2021 Dec 16. 34. ciclo, Anno Accademico 2021.
ANALISI CRITICA DEL DISCORSO LESSICOGRAFICO CINESE: UNO STUDIO DIACRONICO DELLO XIANDAI HANYU CIDIAN 现代汉语词典
This dissertation proposes a study, from a critical and diachronic perspective, of the Xiandai hanyu cidian 现代汉语词典 (hereinafter XHC), a monolingual dictionary of standard modern Chinese (putonghua 普通话) which plays a central role within the lexicographical field of the People’s Republic of China. The compilation of the dictionary is entrusted to the Institute of Linguistics belonging to the Chinese Academy of Sciences and, since its foundation in 1977, to the Chinese Academy of Social Sciences, which are both research institutions placed under direct control of the State Council. Officially published for the first time in 1978, and now in its seventh edition (2016), the XHC was conceived by the political leadership as a tool for promoting the standardisation and the popularisation of the common language, within the broader project for language reform in the 1950s. This research adopts the theoretical framework of critical lexicography, and it is based on the premise that a dictionary can be analysed as a form of discourse and, specifically, as a form of lexicographical discourse, which is also to be understood as being historically specific (Benson 2001). According to this perspective, the contents of language dictionaries and, therefore, their lexicographical discourse, should be considered in relation to the extra-textual reality where their compilation and their fruition take place. They can be seen both as products of their extra-textual reality and as tools playing an active role in the reception, construction, and consolidation, as well as in the dissemination of those meanings that are considered as correct and acceptable by the dominant ideology (Fairclough 1989; Fishman 1995; Hornscheidt 2008, Rodríguez Barcia 2012). The objective of the analysis is to verify whether and to what extent the lexicographical discourse constructed by the XHC reflects and interacts with its extra-textual, ideological, and, therefore, political and social context. The research intends to fill a gap in the critical literature on this dictionary, in which the studies investigating the relationship between lexicographical discourse and ideological elements are, at the present day, still very limited (Lee 2014; Hu H., Hai X. and Hao J. 2019). This dissertation is divided into two main parts: the first consists of the first, the second and the third chapters and the second includes the fourth and the fifth chapters. The first chapter centres upon the definition of the theoretical framework of the study and, specifically, on the relationship between language and ideology, as well as between lexicography and ideology. The second chapter aims firstly at reconstructing the history of the XHC and, subsequently, describing its formal and lexicographical features. The third chapter is dedicated to an evaluation of the state of the art of the research on the dictionary. The fourth chapter presents the methodology constructed and followed for the selection of the dictionary entries taken into analysis. The selection has been carried out through the use of the software AntConc for the analysis of a corpus specifically built for this study. This corpus comprises reports on the work of the government, usually presented to the National People’s Congress by the Prime Minister of the People’s Republic of China, and reports presented by political leaders to the National Congress of the Chinese Communist party (1964-2017). The analysis and the selection of the words (and, therefore, of the dictionary entries) from the political corpus was guided by the definition of both qualitative criteria (morphological and semantic features) and quantitative criteria (number of occurrences). Finally, the fifth chapter is devoted to the analysis, in a critical, ideological, and diachronic perspective, of the definitions of the selected entries, which were collected through the consultation of successive editions of the XHC. Mor
ANALISI CRITICA DEL DISCORSO LESSICOGRAFICO CINESE: UNO STUDIO DIACRONICO DELLO XIANDAI HANYU CIDIAN 现代汉语词典 / C. Bertulessi ; tutor: B. Mottura ; coordinatrice di dottorato: M. V. Calvi. Università degli Studi di Milano, 2020 Jan 24. 32. ciclo, Anno Accademico 2019. [10.13130/bertulessi-chiara_phd2020-01-24].
FROM SYNTHESIS TO APPLICATION:PORPHYRINS AS VERSATILE COMPOUNDS IN ENERGY, SENSING, AND CATALYSIS
Porphyrins, belonging to the class of heterocyclic tetrapyrrolic organic compounds, exhibit notable prevalence in nature. Structurally, these compounds feature a planar arrangement of four interconnected pyrrolic units, forming a flat macrocycle with an extensive conjugated system comprising 18 π-electrons. Their central cavity readily accommodates metal cations, forming stable complexes with transition metal ions. This PhD Thesis delves into the synthesis, characterization, and varied applications of porphyrins. The distinctive colour spectrum of porphyrins, spanning from red to green, originates from robust absorption within the visible spectrum. UV-VIS absorption spectroscopy plays a crucial role in examining their light-harvesting properties. When coupled with electrochemical methods and theoretical calculations, it provides insights into the involved molecular orbitals. Tailoring the chemical structure enables the precise adjustment of the HOMO-LUMO energy gap, resulting in a macrocycle with distinctive absorption and redox properties. This versatility makes it suitable for a wide range of applications, including disease treatment, sensor technology, catalysis, nonlinear optics, and molecular photovoltaics.The synthesis of porphyrins involves conventional approaches such as the Adler-Longo process and Lindsey's one-flask two-step methodology. Challenges in this process include the generation of by-products. Subsequent modifications, including metal coordination and the introduction of functional groups, further refine the porphyrinic core for specific applications.This PhD Thesis presents a comprehensive three-year research project, elucidating the synthesis, characterization, and applications of porphyrins. The exploration encompasses various applications such as Dye-Sensitized Photo-Electrochemical Cells (DSPEC), chemiresistive gas sensors, CO2 reduction catalysis, and second-order Nonlinear Optics (NLO). Each section explores design strategies, synthetic optimization, and performance evaluation, providing an in-depth understanding of the versatile realm of porphyrins.
FROM SYNTHESIS TO APPLICATION: PORPHYRINS AS VERSATILE COMPOUNDS IN ENERGY, SENSING, AND CATALYSIS / C. Albanese ; tutor: F. Tessore ; cotutor: G. Di Carlo ; director of the school: D. Passarella. Dipartimento di Chimica, 2024 May 15. 36. ciclo, Anno Accademico 2023/2024.
INTEGRATED OPTICAL CHARACTERIZATION OF AIRBORNE AND DEPOSITED MINERAL DUST TO IMPROVE THE UNDERSTANDING OF RADIATIVE TRANSFER PROCESSES THROUGH POLAR AND ALPINE SNOWPACKS
The Earth’s cryosphere is undergoing rapid transformation due to climate change, withsignificant reductions in glacier mass and snow cover expected by the end of the century.According to the 2021 IPCC report, mid-latitude glaciers may lose up to 80% oftheir mass, while snow cover could decline by 90%. These changes will disrupt ecosystemsand have profound effects on human activities. Atmospheric aerosols play a crucialrole in climate regulation by interacting with solar and terrestrial radiation. These particlesinfluence the Earth’s radiative energy balance through scattering and absorptionprocesses, affecting cloud formation and precipitation. When deposited on snow andice, aerosols reduce surface albedo, accelerating melting. However, uncertainties persistregarding the full extent of their impact due to incomplete knowledge of aerosol interactionswith visible and near-infrared light. Aeolian mineral dust, in particular, remains anarea of significant research interest due to its uncertain contribution to global radiativebalance. Framed in this context, ice cores from polar and Alpine regions also provideessential historical records of atmospheric composition and climate variability. Chemicaland physical analyses reveal information on greenhouse gases, heavy metals, andmineral dust, aiding climate reconstructions and model improvements. Another criticalresearch area concerns sunlight propagation in snowpacks. Fresh snow exhibits highalbedo, but impurities such as black carbon and mineral dust lower reflectivity, enhancingmelting. Despite numerical modeling efforts, experimental data on light penetrationwithin snow remains limited. To address these knowledge gaps, optical techniques providea non-invasive and non-destructive approach. This research aligns with nationaland international objectives, contributing to improved climate modeling and a betterunderstanding of cryosphere changes.
INTEGRATED OPTICAL CHARACTERIZATION OF AIRBORNE AND DEPOSITED MINERAL DUST TO IMPROVE THE UNDERSTANDING OF RADIATIVE TRANSFER PROCESSES THROUGH POLAR AND ALPINE SNOWPACKS / L. Teruzzi ; tutor: M. A. C. Potenza ; co-supervisor: R. Vecchi ; director of the phd school: A. Mennella. Dipartimento di Fisica Aldo Pontremoli, 2025 Mar 31. 37. ciclo, Anno Accademico 2024/2025.
DEVELOPMENT OF AN INTEGRATED METHODOLOGY TO ASSESS THE PRESENCE AND THE IMPACT OF (MICRO)PLASTICS ON THE RIVER BENTHIC COMMUNITY
Plastic pollution is one of the main environmental issues of the modern society since the development of synthetic polymers with exceptional chemical/physical characteristics in the 20th century led to a continuous production and use of these materials with a great increase in plastic wastes. Consequently, the mismanagement of the end-of-life plastic products induces a constant flux of these materials into the environment, especially in aquatic ecosystems. In this context, rivers are the main collectors of plastics from the continental areas, representing also an important sink of these contaminants in the sediments, with a possible affection of benthic communities.Since most of the studies about plastic pollution in freshwaters primarily deals with water matrix, the main aim of this project was to develop an integrated methodology to monitor the plastic pollution in several freshwater ecosystems, considering the impact of some variables, as the environmental matrix selection and the temporal variation of the contamination, associated to their toxicity evaluation on the benthic macroinvertebrate Chironomus riparius.Firstly, the method setting was crucial to determine the best sampling and analytical setup for plastic monitoring in sediments. This preliminary step was performed in the Lambro River (Northern Italy), which represents an excellent site for the method development due to its high plastic pollution. In detail, plastics were separated from the collected sediments through two density extraction methods, comparing the efficiency of sodium chloride (NaCl) and zinc chloride (ZnCl2) solutions. Results showed a significant difference (p < 0.05) in the extraction performance, with ZnCl2 extracting more plastics (53.3 ± 16.1 plastics/kg sediments wet weight - w.w.) than NaCl (8.3 ± 10.4 plastics/kg sediments w.w.; see manuscript 1).The study moved subsequently to the comparison of plastic pollution in both waters and sediments in four of the main Po River tributaries (Ticino, Adda, Oglio, and Mincio Rivers), using the ZnCl2 solution in the separation processes. Results showed that contamination in sediments was four times higher than in waters, highlighting heterogeneous pollution from both quantitative and qualitative point of view, with the highest levels in Mincio River (62.9 ± 53.9 plastics/m³ in waters and 26.5 ± 13.3 plastics/kg dry weight - d.w. - in sediments) and the lowest in Ticino River (0.9 ± 0.5 plastics/m3 in waters and 6.8 ± 4.5 plastics/kg d.w. - in sediments; see manuscript 2). This study suggested that the monitoring of both matrices provide complementary information: waters offer a picture of an instantaneous contamination, while sediments, representing a sink of plastics, a “historical” pollution. Since waters represent a snapshot of plastic contamination in a water body, the temporal variation of these contaminants in the water matrix was also assessed. Lake Maggiore, from which Ticino River is the main emissary, was selected as reference site due to the highest plastic contamination compared to the other great sub-alpine Lakes. This work revealed a monthly variation in plastic content, from0.02 plastics/m3 in September to 0.29 plastics/m3 in December (see manuscript 3). These findings underscore the importance of comprehensive monitoring plans considering both abiotic matrices (waters and sediments), as well as multi-sampling campaigns throughout the year, to better understand the plastic contamination in freshwater ecosystems.Since the most of studies investigated the toxicity of virgin standard plastic particles, ignoring the wide plethora of shapes, sizes and polymers detected in the environment, in this project the sampled plastics in the abovementioned freshwater lotic environments were used in the exposures of Chironomus riparius. Since no normalized methods for the Chironomid exposure to plastics were available, a modification of the
DEVELOPMENT OF AN INTEGRATED METHODOLOGY TO ASSESS THE PRESENCE AND THE IMPACT OF (MICRO)PLASTICS ON THE RIVER BENTHIC COMMUNITY / R. Sbarberi ; tutor: S. Magni, co-tutor: A. Binelli ; coordinator: M. P. M. Guarino. Dipartimento di Bioscienze, 2025 Apr 07. 37. ciclo, Anno Accademico 2023/2024.
EXPLORING THE DIVERSITY AND FUNCTIONAL PROPRIERTIES OF AKKERMANSIA MUCINIPHILA STRAINS OF HUMAN ORIGIN: A GENOTYPIC AND PHENOTYPIC ANALYSIS
The gastrointestinal (GI) microbiota comprises a complex community of microorganisms that significantly influence the health of their hosts. In recent years, many research efforts have focused on deciphering the role of certain commensal species that seem critical to human well-being. In fact, their presence in the intestine is frequently correlated with healthy hosts and inversely correlated with various pathological states. This has prompted further studies aimed at clarifying whether their use as supplements and/or therapeutics can recapitulate a healthy phenotype.A. muciniphila is one of the few microbiota members that is shown to have an actual causal relationship with improved host health, especially in the context of metabolic disorders. This microorganism, belonging to the phylum Verrucomicrobia, has been described for the first time in 2004 as an anaerobic, Gram-negative bacterium, that colonizes the colonic mucosal layer. A. muciniphila represents one of the main players in this niche acting as mucus degrader, which has been shown to have important implications on host physiology and microbiome composition. Overall, these years' efforts have further strengthened the evidence of the effect of this microorganism on metabolic and immune health by describing casual links and modes of action. For a long time, A. muciniphila represented the only fully described species of the phylum Verrucomicrobia, with the vast majority of studies focused on the type strain, A. muciniphila MucT. Over the past 6 years, large scale genome-based studies have led to the accumulation of evidence that the genus Akkermansia contains other species that likely colonize our gastrointestinal tract, and that even the A. muciniphila species itself is characterized by a wide intra-species genetic heterogeneity. However, how this diversity translates at the phenotypic level and in the relationship with the host is much less investigated. In this context, the isolation and characterization of new strains of A. muciniphila is critical to better understand the physiological diversity and overall health relevance of this species, other than to discover new interesting strains to be further developed as supplements. In this thesis, we have isolated five new strains of A. muciniphila from faecal samples of healthy subjects. The new strains were characterized by performing a series of genotypic and phenotypic analyzes associated with the biology of this species, in order to highlight differences between strains and characteristics that might be relevant in the relationship with the host (e.g. surface properties, anti-inflammatory potential) and related to their safety. Additionally, we worked at the development of an innovative method for the selective detection and quantification of A. muciniphila in multi-species samples based on immunolabeling and flow cytometry.
EXPLORING THE DIVERSITY AND FUNCTIONAL PROPRIERTIES OF AKKERMANSIA MUCINIPHILA STRAINS OF HUMAN ORIGIN: A GENOTYPIC AND PHENOTYPIC ANALYSIS / R. Filardi ; tutor: S. Arioli ; phd dean: D. Mora. Dipartimento di Scienze per gli Alimenti, la Nutrizione e l'Ambiente, 2023. 35. ciclo, Anno Accademico 2022.
BUENOS AIRES: RAPPRESENTAZIONI CULTURALI E RISEMANTIZZAZIONI URBANE DALLA FINE DELLA DITTATURA AL XXI SECOLO
ABSTRACTBetween the end of the last dictatorship (1976-1983) and the early years after the economic crisis of 2001, Buenos Aires undergoes a deep transformation in response to the social fabric erosion perpetrated by the regime (OSZLAK: 2017). This is exacerbated by an assertive privatization policy stemming from adherence to the neoliberal economic model (TORRES: 2001; STANCANELLI: 2002; GORELIK: 2004, 2009, 2013; CICCOLELLA; BAER: 2008; PÍREZ: 2009).Consequently, specific neighborhoods and urban spaces experience a redefinition, witnessing substantial changes in the forms and locations of social aggregation (CANCLINI: 1995; WORTMAN: 2007, 2009). Ethnic barrios emerge, inhabited by new Latin American and Asian migrants, while villa miseria and informal settlements proliferate, often juxtaposed with opulent country clubs and torres jardín, enclosed and monitored residential areas (TORRES: 2001, 2006; GORELIK: 2004, 2009; 2015; PRÉVÔT SCHAPIRA: 2002). Following the 2001 crisis, the capital also witnesses the rise of self-managed initiatives by civil society in response to the economic and social crisis, contributing to the city's redefinition (SARLO: 1994; LECHNER: 2000; ROSBOCH: 2006; CECCONI: 2009). This entails a process of reshaping the use and practices of the territory (SARLO: 1994; LECHNER: 2000; ROSBOCH: 2006; CECCONI: 2009). Additionally, from 2003 onward, during Néstor Kirchner's presidency, Buenos Aires evolves into a stage for a new definition, focused on the reconstruction of the memory of dictatorship crimes (FÈVRE: 1993; SILVESTRI: 1999; JELIN: 2002; CATELA DA SILVA: 2019; GUGLIELMUCCI, LÓPEZ: 2019; GUGLIELMUCCI: 2011; SAID: 2011).Different cultural expressions give voice to individuals inhabiting this new urban landscape: the desaparecidos, absent yet omnipresent; the insiliados, reclaiming their voice after a prolonged silence; survivors returning from exile seeking to reclaim their city; Latin American migrants, alongside diverse sub-proletarians (cabecitas negras, villeros, indigenous) and those facing severe poverty due to the crisis.This research examines a corpus of literary works and songs wherein the city assumes a central role, aiming to offer a comprehensive overview of representations of Buenos Aires. It establishes a dialectical relationship between cultural expressions subject to diverse sectoral studies or not always considered by academic criticism. Furthermore, through the analysis of two self-managed cultural initiatives - the Pachamama Cultural Center and the Club Atlético Fernández Fierro - the study highlights the efficacy of such experiences.The thesis is structured into five chapters. The first chapter paints a portrait of the city through «palabras y sonidos» (PUJOL: 1993), literary and musical texts that have represented the city throughout history until 1983, emphasizing the extensive literary and musical tradition associated with Buenos Aires. The second chapter outlines the historical context of the city from the restoration of democracy to the early years after the 2001 crisis, considering the trauma of dictatorship, mourning, and the realization of the neoliberal model's failure. The third chapter analyzes two self-managed cultural centers, the Pachamama Cultural Center and the Club Atlético Fernández Fierro, as case studies. The fourth chapter focuses on a broad corpus of literary texts, exploring the concept of civilibarbarie introduced by Elsa Drucaroff (2017) and the relationship between political events and literary production. In the fifth chapter, a vast corpus of song lyrics belonging to the three most representative genres during the considered period - tango, rock, and cumbia - is examined. The thesis aims to provide a comprehensive image of a city deeply marked by dictatorship and neoliberalism yet functioning as a fertile cultural laboratory capable of redefining the concepts of center and periphery, margin a
BUENOS AIRES: RAPPRESENTAZIONI CULTURALI E RISEMANTIZZAZIONI URBANE DALLA FINE DELLA DITTATURA AL XXI SECOLO / M. Fumagalli ; tutor: I. M. Bajini ; coordinatore: L. Pinnavaia, M. V. Calvi. - Università degli Studi di Milano. Dipartimento di Beni Culturali e Ambientali, 2024 May 27. 36. ciclo, Anno Accademico 2023.
THE POLITICS OF SANCTIONS COMPLIANCE. EXPLAINING SANCTIONS EFFECTIVENESS THROUGH THE LENS OF DOMESTIC VETO PLAYERS. EMPIRICAL EVIDENCE FROM THE CASES OF ERITREA, IRAN, AND SUDAN.
Sanctions scholarship overwhelmingly expects that authoritarian states defy and resist the demands attached to sanctions imposed on them. In practice, however, authoritarian states do sometimes accommodate to sanctions-related demands and offer political concessions consistent with them. What explains their choice of compliance or defiance? When do they decide to offer political concessions aligned with sanctions-related demands, and why are those concessions sometimes short-lived and not robust, and why can they be more or less expedited? This research project argues the answer lies in the domestic configuration of institutional and political constraints, also called veto players, of the states under sanctions. As a contribution to the domestic-politics branch of the literature on international sanctions, which has become increasingly more relevant in light of the evolution of sanctions from comprehensive to targeted, this research project aims to investigate the under-researched application of a typically public-policy approach such as the veto player analysis to the literature on international sanctions and understand if and how domestic constraints such as veto players, who cut across regime type classifications that are frequently –but often inconclusively– used in the literature on sanctions, affect the effectiveness of sanctions in achieving the desired political concessions. Differently from previous studies, this project intends to examine this interaction along three different dimensions of the targeted state’s aggregate policy response to sanctions, that are the direction, robustness, and expedition of the policy change which together describe the overall effectiveness of sanctions. To test how three different elements of the configuration of the targeted state’s veto players such as the concentration of the agenda power, the congruence of policy preferences, and the internal cohesion, can affect those three outcome dimensions, the project adopts different methodological strategies. After performing some statistical analyses which expose some issues which remain largely unaddressed by the extant literature and probe alternative hypotheses, the project builds on those findings to develop a new theoretical framework and related spatial model which is applied to few selected comparative case studies to trace how veto power operates inside a state targeted by sanctions and test the hypotheses identified, one per dimension of the policy response to sanctions. This empirical part aims to verify the empirical support for the hypotheses and illustrate how the explanatory power of the veto player lens outperforms explanations that focus instead only on regime type classifications in decoding the politics of sanctions compliance.
THE POLITICS OF SANCTIONS COMPLIANCE. EXPLAINING SANCTIONS EFFECTIVENESS THROUGH THE LENS OF DOMESTIC VETO PLAYERS. EMPIRICAL EVIDENCE FROM THE CASES OF ERITREA, IRAN, AND SUDAN / T. Corda ; tutor: A. Carati ; coordinatore del dottorato: M. Jessoula. Dipartimento di Studi Internazionali, Giuridici e Storico - Politici, 2022 Apr 13. 34. ciclo, Anno Accademico 2021.
RABPHILIN3A: FROM NMDA RECEPTOR MEMBRANE DYNAMICS TO NEURODEVELOPMENTAL DISORDERS
Rabphilin3A (Rph3A) is a synaptic protein essential for regulating exo- and endocytosis processes at presynaptic terminals and dendritic spines. It promotes the stabilization of the NMDA-type glutamate receptors (NMDARs) at the cell surface, forming a ternary complex with the PSD-95 and GluN2A subunit of the receptor. This complex is needed for long-term potentiation (LTP) induction and NMDAR-dependent hippocampal behaviors, such as spatial learning. Under physiological conditions, Rph3A is present in approximately 50% of hippocampal spines. Notably, Rph3A positive (Rph3A+) spines exhibit enlarged spine head and postsynaptic density size, indicating enhanced synaptic stability. LTP induction increases both the number of Rph3A+ spines and NMDAR synaptic retention, thus indicating that plasticity promotes the postsynaptic localization of the protein. Importantly, the disruption of Rph3A/NMDAR complex impairs GluN2A accumulation at postsynaptic membrane, preventing LTP induction. Our group has also previously demonstrated that overexpression of Rph3A in primary hippocampal neurons is sufficient to occlude the LTP-induced formation of new dendritic spines, confirming a key role for the Rph3A/GluN2A pathway in these processes. Given these premises, the first aim of my thesis was to determine whether Rph3A serves as a molecular marker for potentiated synapses. Using calcium imaging in primary hippocampal neurons, I found that overexpression of Rph3A leads to significantly increased postsynaptic calcium influx. To assess the role of Rph3A at the level of individual synapses, I performed single-spine glutamate uncaging in live neurons. Spines overexpressing Rph3A failed to undergo further potentiation upon stimulation, suggesting that they were already in a potentiated state. Additionally, I investigated how Rph3A influences the dynamics and synaptic localization of NMDARs. In the second part of my thesis, I explored the pathological consequences associated with RPH3A gene. By combining trio-based exome sequencing, GeneMatcher data, and analysis of the 100,000 Genomes Project, our collaborators identified six heterozygous missense variants in RPH3A gene. Four cases had a neurodevelopmental disorder (NDD) with untreatable epileptic seizures [p.(Gln73His)dn; p.(Thr450Ser)dn; p.(Arg209Lys); p.(Gln508His)], and two cases [p.(Asn618Ser)dn; p.(Arg235Ser)] showed high functioning autism spectrum disorder (ASD). Using primary hippocampal neurons, I demonstrated that four of these variants (p.(Arg209Lys), p.(Thr450Ser), p.(Gln508His), and p.(Asn618Ser)) disrupted the synaptic co-localization of PSD-95 and GluN2A and led to increased surface levels of GluN2A at extrasynaptic sites. In addition, RPH3A mutants induce defects in NMDAR activity at synaptic sites, affecting postsynaptic calcium and glutamate influx at dendritic level. Collectively, this PhD work identifies Rph3A as a molecular marker of potentiated synapses and a critical regulator of synaptic NMDAR localization. Moreover, it provides the first evidence that missense gain-of-function variants in RPH3A can disrupt synaptic function and contribute to a spectrum of neurodevelopmental disorders, from untreatable epilepsy to autism spectrum disorder.
RABPHILIN3A: FROM NMDA RECEPTOR MEMBRANE DYNAMICS TO NEURODEVELOPMENTAL DISORDERS / M. Barzasi ; tutor: F. Gardoni ; coordinatore: G. D. Norata. Dipartimento di Scienze Farmacologiche e Biomolecolari Rodolfo Paoletti, 2025. 38. ciclo, Anno Accademico 2024/2025.
VALORIZATION OF BY-PRODUCTS FROM AGRI-FOOD SUPPLY CHAINS FOR THE DEVELOPMENT OF FUNCTIONAL INGREDIENTS AND NUTRACEUTICAL
The integration of sustainable practices, advanced analytical methodologies, and the valorization of bioactive compounds derived from agri-food supply chains is crucial for addressing global challenges in health, industry, and environmental management. This doctoral research explores the potential of agro-industrial by-products, focusing on their transformation into valuable resources through eco-friendly processes and state-of-art analytical characterization. A significant component of this research is the study of bergamot by-products. A scalable, eco-friendly extraction process was developed to obtain phytochemically complex materials rich in flavonoids and other bioactive compounds, such as stachydrine. This bergamot-derived extract demonstrated promising anti-inflammatory and antioxidant properties, which resulted to be helpful in treating obesity in an animal model and related physiological dysfunctions, offering a sustainable and efficient approach to valorize industrial residues. By avoiding conventional chemical purification steps and focusing on preserving bioactive compounds, this research highlights the potential of bergamot in nutraceutical applications, including the prevention of oxidative stress and inflammation-related conditions. The study also focuses on the valorization of white grape skins (WGS), a by-product of grappa production, which were transformed into VITUVA®, an industrial-scale extract with a balanced polyphenol profile. Employing green and water-based extraction methods, the study underscores the scalability, safety, and environmental compatibility of this approach. Advanced LC-HRMS profiling identified metabolites such as flavonoids and phenolic acids, linked to the extract's significant anti-inflammatory activity (via NF-κB modulation) and antioxidant properties, despite a limited NRF2 activation. These findings highlight the dual potential of VITUVA® in addressing inflammation and oxidative stress, opening the way for its application in the nutraceutical field and cosmetics. In parallel, this research investigates the parahormetic mechanisms underlying the health-protecting effects of grape seed procyanidins. These compounds demonstrated a remarkable capacity to modulate redox signaling and mitigate oxidative damage through the conversion into species, such as 5-(3′,4′-dihydroxyphenyl)-γ-valerolactone, that acts as electrophiles and thus activating Nrf2 pathway. By focusing on their hormetic effects, this study highlights the delicate balance between beneficial and potentially adverse responses at different doses, a crucial aspect for future therapeutic applications. Additionally, pre-ADMET evaluations of 5-(3′,4′-dihydroxyphenyl)-γ-valerolactone, the main bioactive metabolite derived from proanthocyanidins consumption, provided valuable insights into its pharmacokinetic profile, safety, and bioactivity, further supporting the potential in nutraceutical and pharmaceutical contexts of proanthocyanidins.Overall, this doctoral research bridges the principles of circular economy, green chemistry, and advanced omics techniques, demonstrating how the transformation of industrial by-products into high-value compounds can drive innovation while reducing environmental impact. By integrating environmental oversight with scientific discovery, the findings contribute to a sustainable model for industrial processes and pharmaceutical development. Future directions involve not only exploring the application of products such as VITUVA®, grapeseed procyanidins and bergamot-derived extracts in cosmetic formulations and nutraceuticals but also continuing studies to elucidate the mechanisms of action of bergamot compounds and grape-derived polyphenols. These efforts aim to definitively establish their role in advancing the circular economy and sustainability within the domain of raw materials for nutraceutical and cosmetic use, highlighting their phy
VALORIZATION OF BY-PRODUCTS FROM AGRI-FOOD SUPPLY CHAINS FOR THE DEVELOPMENT OF FUNCTIONAL INGREDIENTS AND NUTRACEUTICAL / L. Della Vedova ; tutor: G. Aldini ; co-tutor: G. Baron ; coordinatore: G. Vistoli. Dipartimento di Scienze Farmaceutiche, 2025 May 09. 37. ciclo
JNKS AS THERAPEUTIC TARGETS TO TACKLE SYNAPTIC DYSFUNCTION IN NEURODEVELOPMENTAL AND NEURODEGENERATIVE DISEASES
Brain disorders are the first leading cause of disability and the second leading cause of death worldwide. Decoding the mechanisms of brain diseases is mandatory to improve diagnosis, neuroprotective strategies, and treatments.Although there are many different pathological and clinical manifestations, as well as causes and molecular pathways underlying neurological disorders, increasing evidence suggest that synapses, having a crucial role in neuronal communication and brain functions, are the first station that degenerates and loses functionality in neurological diseases.This concept has led to the theory of brain disease, from neurodevelopmental to neurodegenerative, as synaptopathies, in which the synaptic impairment is a shared pathogenic feature.The main aim of this thesis was to dissect this concept, focusing on a specific pathway particularly involved in brain functions and dysfunctions: the c-Jun N-terminal Kinase (JNK) signalling pathway. More in details, we studied the role of JNK in two neurodevelopmental syndromes, Rett and Angelman, and in a chronic brain illness, Alzheimer disease, focusing on the synaptopathy as the first initial mechanism of many different brain diseases.In Rett Syndrome, a rare severe developmental disease, we studied the synaptic dysfunction of two different murine models identifying JNK as an important actor downstream MECP2, the gene mutated in the pathology. We demonstrated that the specific inhibition of JNK, by D-JNKI1 treatment, strongly improved the symptoms and the molecular disorganization of the PSD region in both mice models. Then, we proved JNK activation in human neurons, differentiated from human MECP2-mutated iPSCs (MECP2mut), compared to the isogenic control expressing wild-type MECP2 allele (MECP2wt). The JNK signal was activated in the MECP2-mutated iPSCs and not in control iPSCs, and D-JNKI1 blocked the MECP2mut -induced neuronal death.In Angelman Syndrome, we analysed the synaptic dysfunction in the UBE3A+/- mouse model. JNK was strongly activated in the brain of these mice, suggesting its important role also in this disorder. The D- JNKI1 treatment improved the behavioural defects, and this correlated with the stabilization of the synaptic biomarkers.Finally, we focused on the Alzheimer’s synaptic dysfunction, the most characterize synaptopathy, to study JNK role in a chronic illness.Alzheimer mice model 5XFAD presented a powerful JNK activation together with synaptic dysfunction and cognitive impairment. We then centre our attention on the selective brain JNK isoform, JNK3, the most responsive to stress stimuli, finding that 5XFAD mice presented a powerful increase of JNK3 levels. Therefore, we decided to test the potential neuroprotective effect of the specific inhibition of JNK3. In fact, thanks to the collaboration with Prof. Falconi, we have in the laboratory the specific JNK3 inhibitor, dSIMBA2. Before testing this in-vivo, we studied the effect of JNK3 inhibition in an in-vitro model of synaptic dysfunction induced by Ab oligomers (ABO) to define the right dose and the neuroprotection against ABO-induced synaptic dysfunction, finding that dSIMBA2 prevents ABO toxicity and the induced-synaptopathy.5To summarize, the data obtained in this thesis participate in the reinforcement of JNK as a central actor in the degeneration mechanisms of the synapses that characterized both neurodevelopmental and neurodegenerative diseases. In addition, this work adds new elements to define JNK3, the most responsive JNK isoform to stress, as a key player in the AD, and an important target for the treatment of synaptic injury, potentially also in other brain illnesses. Developing new compounds able to inhibit one of the most responsive kinases to stress stimuli may be an intriguing field to explore to prevent synaptic dysfunction.
JNKS AS THERAPEUTIC TARGETS TO TACKLE SYNAPTIC DYSFUNCTION IN NEURODEVELOPMENTAL AND NEURODEGENERATIVE DISEASES / C.a. Musi ; tutor: T. Borsello ; coordinator: G. D. Norata ; reviewer: M. Boido ; reviewer: M. Courtney. Dipartimento di Scienze Farmacologiche e Biomolecolari, 2022 Apr 04. 34. ciclo, Anno Accademico 2021.
UN ESTATUTO COMÚN DE DERECHOS PARA LOS SOLICITANTES DE PROTECCIÓN INTERNACIONAL EN LA UNIÓN EUROPEA
For decades the European Union has developed a regulatory system that has no equal on the international scene: the Common European Asylum System ( hereinafter CEAS) in order to achieve the objective enshrined in Article 78 of the TFEU: to grant an adequate protection status to any third-country national in need of protection. However, despite its many achievements, the CEAS remains far from being perfect. Following the massive migratory influx that reached Europe between 2015 and 2016, it became evident that the EU's international protection policy was deficient; and the consequence of its failures was a succession of grave violations of the international protection seekers' rights. Many used the collapse of overburdened national systems to justify the mistreatment of thousands of applicants for protection. This served the EU and its states to justify a controversial policy of externalization of migration controls and the adoption of dissuasive measures as varied as they were questionable. However, such violations of the rights of applicants were already occurring years before, when the number of arrivals was much lower.Following a study of the CEAS, it has been observed that it suffers from shortcomings that make it an inefficient and non-solidarity-based system, which provides an insufficient degree of harmonization of international protection procedures and conditions related to the reception of applicants. Thus, due to a plurality of procedures, deadlines, guarantees and rights, the recognition rates and reception conditions granted by one State or the other can make a transcendental difference for the applicant. For all these reasons, the legal status of the applicant for protection is affected by great legal uncertainty, which undoubtedly weakens their protection.Consequently, the incorporation of a new regulation to the CEAS is proposed: A regulation concerning the common statute of rights of the applicant for international protection in the European Union. It is argued that it can constitute a normative tool against the systematic violations suffered by a particularly vulnerable group, favouring the efficacy of the system. Thus, this study analyses the current state of the CEAS, the reasons that support the proposal, and its objectives, concluding with an explanation of the guidelines that should be developed in the regulation.
UN ESTATUTO COMÚN DE DERECHOS PARA LOS SOLICITANTES DE PROTECCIÓN INTERNACIONAL EN LA UNIÓN EUROPEA / M.d. Requena De Torre ; tutor: F. Scuto ; tutor: J.M. Porras Ramírez (Granada) ; coordinatore del Dottorato: F. Biondi ; coordinadora del doctorado (Granada): F. Villalba Pérezcoordinadora del doctorado (Granada): F. Villalba Pérez. Dipartimento di Studi Internazionali, Giuridici e Storico - Politici, 2023 Oct 25. 35. ciclo, Anno Accademico 2022.
STUDY OF THE IMMUNE SYSTEM IN TWO LIVESTOCK SPECIES OF VETERINARY INTEREST
AbstractPathogenic microorganisms employ various strategies to replicate, spread, and pose threats to host functions. Meanwhile, the immune system's task involves eliminating these harmful microbes and toxins while avoiding excessive damage to self-tissues and beneficial microbiota. This complex task necessitates a diverse array of protective mechanisms. In mammals, the immune system comprises both innate and adaptive components, working together to respond to microbial infections. A comprehensive understanding of the host immune system is crucial for advancing vaccine and prevention strategies, as well as diagnostic markers. In light of these considerations, my PhD thesis investigates different aspects of the immune system with an emphasis on my latest discoveries in two pivotal projects. The virulence factors of pathogens play a crucial role in determining the adaptation of bacteria to specific host niches. Enteric colibacillosis is a commonly occurring illness and cause of death globally in pigs that are being nursed or weaned. F4 (K88) or F18 adhesive fimbriae are involved in enterotoxigenic (ETEC) infections, leading to post-weaning diarrhoea (PWD). They mediate bacterial binding on enterocytes of the small intestine. After colonisation, F4 and F18 E. coli strains produce enterotoxins that can cause local or systemic effects. These enterotoxins induce secretory diarrhoea, leading to severe clinical signs, reducing production performances and increasing use of drugs. Vaccination of pregnant sows with inactivated E. coli stimulates the production of specific antibodies passed into colostrum and milk for the protection of suckling piglets against ETEC diarrhoea. However, passive protection is rapidly lost after weaning, and newborns can rapidly become susceptible to disease. At weaning, triggering gut mucosal immunity with local production of IgA antibodies by secreting B cells (SIgA) could represent a valid approach to prevent PWD. The present study aims to clarify the immunogenicity of a new oral inactivated F4/F18 vaccine for active immunisation of piglets against PWD caused by F4-ETEC or F18-ETEC by ELISA and ELISPOT assay. Furthermore, some pigs can be genetically resistant to ETEC due to the lack of specific receptors in the gut for fimbriae. It is important to consider the piglet's genotype when interpreting the immunogenicity of the vaccine. This could affect the outcome of the main objective of the project, which is to evaluate the effectiveness of the oral vaccine. To determine whether a pig is susceptible or resistant to F4/F18, I applied a genotyping protocol based on PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism). Two heat-inactivated ETEC strains along with human-interferon-alpha (IFNα) as adjuvant were administrated to piglets born from unvaccinated sows. Piglets were divided into vaccinated and control groups, and various samples were analysed using ELISA for anti-F4/F18 IgA and IgG levels. Specific F4/F18 immunoglobulins (IgG and IgA) were found in sow colostrum regardless of vaccination, suggesting E. coli circulation in the farm. Results revealed elevated anti-F4/F18 IgA levels in vaccinated piglets' faecal and saliva samples post-vaccination and significant differences in serum IgA levels between vaccinated and control groups and within the control group. The control group had higher levels of maternal-derived anti-F4 IgG, indicating potential pathogen circulation. The pregnant sows, each at day 70 of gestation, were obtained from a conventional herd in which there had been no clinical outbreaks of E. coli enteritis in post-weaning piglets. The selection of sows (from a conventional farm instead of from an SPF facility) was in line with our goal to create realistic field conditions, ensuring the vaccine's effectiveness in a typical farm environment where E. coli exposure is common. Once the animals arrived at the IZSLER a
STUDY OF THE IMMUNE SYSTEM IN TWO LIVESTOCK SPECIES OF VETERINARY INTEREST / A. Inglesi ; tutor: F. Riva;coordinatore: F. Ceciliani. - Lodi. Dipartimento di Medicina Veterinaria e Scienze Animali, 2025 Feb 25. 36. ciclo, Anno Accademico 2022/2023.
THE SPINTERFACE PROBLEM: AB-INITIO STUDIES OF FE-PHTHLOCYANINE ON TRANTITION-METAL OXIDES.
At the interface between a transition-metal oxide and a metal-organic molecule, electrons form interesting hybrid electronic states that characterize what is commonly called an organic spinterface. The central idea of our study is the possibility of exciting a spin coherent wave inside the substrate through a light-induced excitation of the molecule - in other words, actively perturbing the spinterface to modify its magnetic state. In this framework, the spinterface is studied for its electronic, magnetic and optical properties through ab-initio approaches. Specifically, we selected the transition-metal oxides: NiO(001), CoO(001) and Cr2O3(0001) as substrates. These materials are antiferromagnetic at high and/or room temperature and are characterized by low spin-orbit interaction, making them promising candidates for spin-excitation transport. For the metal-organic molecule, we chose Fe-phthalocyanine, a versatile species valued for its adaptability across various applications.In addition to the insights specific to each molecule/substrate combination revealed by our ab-initio simulations, three main findings emerged regarding the goal of activating the spinterface by light. First, significant hybridization between the molecule and the substrate is advantageous for maximizing the transfer of the light-induced molecular excitation onto the magnetic couplings between substrate spins. Second, to impact the substrate's magnetic couplings, this hybridization needs to be spin-unbalanced. In this regard an adsorption configuration breaking the spin symmetry of the substrate is sufficient, so that the adsorption of a magnetic molecule is not required. Third, effective control over spinterface tunability requires a distinct retention of molecular and substrate character: hybridization should not be so strong that they become indistinguishable. In terms of light excitation spectra, this translates into high-intensity transitions between states with identifiable molecular and substrate characteristic.
THE SPINTERFACE PROBLEM : AB-INITIO STUDIES OF FE-PHTHALOCYANINE ON TRANSITION-METAL OXIDES / M. Marino ; supervisor: G. Fratesi ; co-supervisor: S. Achilli ; coordinator: A. Mennella ; external referee: M. Cococcioni ; external referee: P. Puschnig. , 2024 Dec 22. 37. ciclo, Anno Accademico 2024/2025.
WHEN AESTHETICS MEETS ETHICS: THE SOUTH AFRICAN SHORT STORY IN ENGLISH, 1920-2010
This thesis explores the development of the short-story genre in English in South Africa from the late 1920s to the present day. I intend to close read a corpus of short stories by eight South African writers (Rolfes and Herbert Dhlomo, Peter Abrahams, Can Themba, Alex La Guma, Mtutuzeli Matshoba, Ahmed Essop, and Zoë Wicomb) with particular focus on the dialogue between ethics and aesthetics performed by these texts vis-à-vis the evolution of South Africa’s socio-political situation (before, during, and after apartheid). Often considered a minor genre, the short story actually thrives in post-colonial contexts, like South Africa, where the writing and publishing conditions available to writers can be under a set of material constraints; the brevity of the short story, however, allows for swifter conditions of production and circulation than the ones required by the novel. Consequently, my analysis also focuses on the various publishing venues of these short stories, in particular local newspapers and little magazines, considered as pivotal spaces for the intersection between ethics and aesthetics, which often results in a cross-fertilization between journalism and the short story. Negotiating the different tensions of literature, politics, journalism, and commercial interests, the shifting contexts and editorial policies of newspapers and magazines represent particularly interesting case studies from which the unstable nature of the definition of the ‘literary’ emerges, which is never detached from a political dimension.This study also seeks to expose the fictional and ideological nature of the ‘poetics of authenticity’ that frequently characterises the selected short stories, which display (and often play with) the conventions of realism to achieve a certain patina of authenticity. The poetics (and politics) of authenticity is, I believe, the most significant feature of the interdependence of ethics and aesthetics in the South African short stories in English analysed in this thesis. Between the late 1920s and the early 1980s – that is, before and during apartheid – claims to authenticity mark, albeit in different ways and through different fictional strategies, the selected short stories. This reflectionist trend, which is always a literary construct, responds to the ethical imperative of documenting the lives of the Black population under a segregationist regime, but it is also influenced by the available publishing venues (newspapers and magazines). As the South African short story in English progresses in its post-apartheid phase, essentialist, fixed notions such as ‘authenticity’ are exposed as a fictional construct by authors such as Zoë Wicomb. Ultimately, this study aims to foreground the impossibility of separating the literary (both the text and its materiality) from the political.
WHEN AESTHETICS MEETS ETHICS: THE SOUTH AFRICAN SHORT STORY IN ENGLISH, 1920-2010 / M. Fossati ; tutor: G. Iannaccaro ; co-tutor: A. Van Der Vlies ; coordinatore: M. Calvi. Dipartimento di Lingue, Letterature, Culture e Mediazioni, 2022 May 05. 34. ciclo, Anno Accademico 2021.
A GUT PATCH FOR OBESITY - SUPERABSORBENT HYDROGEL AS A NOVEL THERAPEUTIC INTERVENTION
The intestinal barrier is the first line of defense against pathogens that separates our body from the external environment. Its function is to maintain intestinal tissue homeostasis, regulating the passage of molecules and protecting from harmful microbes. This barrier is made up by the mucus layer, critical for limiting the exposure to the gut microbiota, and by intestinal epithelial cells sealed by inter-epithelial tight junctions. A strong cause-effect relation between intestinal permeability alterations and both intestinal and systemic diseases (obesity, type 2 diabetes and NAFLD) is becoming evident, suggesting that restoring proper gut permeability by targeting the intestinal epithelial barrier could represent a successful intervention. The present study proposes the use of a naturally-derived superabsorbent hydrogel to restore gut permeability, likely acting on the intestinal epithelial barrier, either directly or indirectly. In a mouse model of HFD-induced obesity, hydrogel diet supplementation prevented early development of adiposity, hepatic steatosis and intestinal barrier perturbations, mainly reinforcing intestinal epithelial TJ and inhibiting bacterial LPS translocation and endotoxemia. In long-term HFD experiments, both prophylactic and therapeutic hydrogel administration reduced body weight, adipose tissue deposition and improved metabolic parameters; notably, this was paralleled by the restoration of intestinal barrier permeability, thus limiting endotoxemia, as well as the reshaping of the gut microbiota composition despite dysbiosis associated to HFD feeding. Indeed, both dysbiosis and low-grade inflammation are hallmarks of metabolic diseases.Collectively our data show the potentiality of this superabsorbent hydrogel as a completely naturally-derived and non-invasive therapeutic tool for the treatment of obesity and metabolic disorders.
A GUT PATCH FOR OBESITY - SUPERABSORBENT HYDROGEL AS A NOVEL THERAPEUTIC INTERVENTION / A. Silvestri ; tutor: M. Alcalay ; co-tutor: M. Rescigno ; internal advisor: D. Pasini ; external advisor: A. Sannino ; coordinator: G. Viale. Dipartimento di Scienze della Salute, 2020 Dec 11. 32. ciclo, Anno Accademico 2020. [10.13130/silvestri-alessandra_phd2020-12-11].
A TRIPLE FLORIGEN SYSTEM IS ESSENTIAL FOR FLOWERING AND PANICLE ARCHITECTURE IN RICE
The transition from a vegetative phase to a reproductive one has always been deeply studied in plants and in particular in the cereals due to the relevance of this trait for seed yield. The transition occurs upon the arrival of a mobile flowering signal, the florigen, at the shoot apical meristem (SAM).In rice (Oryza sativa), there are two different florigens: Heading Date 3a (Hd3a), that promotes flowering in short day (SD) conditions and Rice Flowering Locus T 1 (RFT1) that promotes flowering in both short and long- days (LD). They are transcribed and translated in leaves once environmental conditions are optimal, and then transported through the phloem to the SAM. After reaching the shoot they form a heterohexameric complex named Florigen Activation Complex (FAC) binding to two 14-3-3 proteins and to two bZIP transcription factors and activate conversion of the shoot apex into an inflorescence by promoting expression of inflorescence- identity genes.Here, we show that Hd3a and RFT1 arrival at the SAM activates FLOWERING LOCUS T-LIKE 1 (FT-L1), encoding a third florigen-like protein showing atypical characteristics.Expression data from qRT-PCR, in situ hybridization and transcriptional marker lines indicated that its mRNA is mainly expressed in the SAM and persists during all developmental stages of inflorescence development. Misexpression of FT-L1 from meristem- and tissue-specific promoters indicates that only when expressed in the SAM, FT-L1 has florigenic activity.Isolation and analysis of EMS and CRISPR mutants showed that FT-L1 strengthens the effects of Hd3a and RFT1 during conversion of the vegetative meristem into inflorescence, in an addictive manner. Furthermore, it controls panicle branching by inhibiting secondary branching and promoting spikelet meristem differentiation.Thus, we propose a triple florigenic module that promotes a balanced progression of inflorescence identity and determinacy.
A TRIPLE FLORIGEN SYSTEM IS ESSENTIAL FOR FLOWERING AND PANICLE ARCHITECTURE IN RICE / F. Giaume ; scientific supervisor: F. Fornara ; phd school coordinator: M. Kater ; revisore: G. Valè, S. Boden. Dipartimento di Bioscienze, 2022 Jul 21. 34. ciclo, Anno Accademico 2021.
PHOTOPERIODIC REGULATION OF RICE STEM ELONGATION BY THE GENE PINE1
In several plant species, inflorescence formation is accompanied by stem elongation. While these two processes need to progress harmoniously, until recently no genetic link had been found that could integrate and coordinate both pathways. Only a recent study has shed light on a new and unexplored pathway that, via the PREMATURE INTERNODE ELONGATION 1 (PINE1) gene, links flowering with stem elongation [1]. PINE1 is a zinc finger transcription factor able to repress stem elongation during vegetative growth. During floral induction, the florigenic signal can silence PINE1 expression, triggering internode elongation. This thesis aims to explore the mechanism of action of PINE1 and the potential exploitation of this pathway for rice breeding for agriculture. The hypothesis that PINE1 could be regulated by genes involved in the flowering machinery has been tested by generating overexpressors of the known flowering related genes OsMADS14, OsMADS15 and FTL1 to assess their phenotype regarding stem elongation. Indeed, FTL1, the expression of which is promoted by florigens, could be involved in PINE1 inhibition during flowering. Previous literature on genes similar to PINE1 reported interaction with co-repressors proteins. Thus, some of the recurring interactors were cloned to test whether they can effectively interact with PINE1. The data suggests that PINE1 is able to form a complex with TOPLESS (TPL) proteins, which may function in transcriptional repression through chromatin remodeling. The recognition of PINE1 by TPL depends on the presence of EAR-motifs, which are also important for PINE1 function overall. As PINE1 is a regulator of stem elongation and plant height is an important breeding trait, we attempted to modify plant height by modulating PINE1 expression by introducing mutations in its promoter region. Some of the generated plants did show strong dwarfism, indicating that some mutation in the promoter region lead do a PINE1 overexpression. For the same purpose of finding ways to control plant height, we also identified some candidate genes controlled by PINE1 using RNA sequencing and created knock-out mutations using the CRISPR/Cas9 system to further explore their role in stem elongation.Overall, this work establishes the framework for a new pathway that controls plant architecture and has implications for crop improvement. Further studies will be needed in the coming years to elucidate the entirety of this cascade of events from flowering to stem elongation and to explore the potential applications of this research for agriculture.
PHOTOPERIODIC REGULATION OF RICE STEM ELONGATION BY THE GENE PINE1 / G. Vicentini ; tutor: S. Pierce, V. Brambilla ; coordinatori: D. Bassi, P. A. Bianco. Dipartimento di Scienze Agrarie e Ambientali - Produzione, Territorio, Agroenergia, 2023 Jun 28. 35. ciclo, Anno Accademico 2022.
INTERPLAY BETWEEN RAW MATERIAL AND PROCESSING CONDITIONS FOR THE FORMULATION OF PULSE-BASED PRODUCTS
As modern consumers are becoming increasingly conscious about health and sustainability, theyare demanding plant-based tasty and convenient food products. In this context, using pulses infood formulations has strongly grown owing to their agronomic, nutritional and health benefits. Inthe scenario of reformulating food products with pulses, extrusion plays a key role as it can leadto a wide range of products such as snacks, pasta, and meat analogues. Although some studiesprovided information on the suitability of pulse extrusion, it is still unclear which pulse performsbetter for each type of application and how to perform the process to obtain products withenhanced quality.This project focused on understanding the relationship between raw material, processingconditions and product quality, with the final goal of identifying the most suitable combination oftype of pulse and extrusion process for the production of extruded products. Three extrudedproducts were chosen as representative of the extrusion carried out at different moisture levels:snacks (10% moisture), pasta (30% moisture), and meat analogues (50% moisture).As regards snacks, a literature review was necessary to frame the industry (chapter 4.1.1). Thisrevealed the lack of information regarding the use of pulses (as whole or fractions) in extrudedsnacks. Therefore, the research focused on the use of bran from chickpeas and green peas (at15% and 30% enrichment level) on the chemical and physical characteristics of snacks (chapter4.1.2). Although the enrichment with pulse bran affected the snack features, the developedproducts were preferred by the consumers compared to the control. These results lead to a focuson sensory aspects (chapter 4.1.3), highlighting that crumbliness and mild flavor positivelyinfluenced hedonic scores, whereas stickiness, dryness, hardness, and to a lesser extent visualaspect, affected them negatively. Comparing the extrusion aptitude of chickpeas, green peas,and red lentils, the latter gave the best results for both snacks and pasta products (chapter 4.1.4).In the case of pasta, the literature review showed great interest in using unconventional rawmaterial (chapter 4.2.1). A picture of the pulse pasta market highlighted differences according tothe type of pulse and brand (chapter 4.2.2), suggesting further investigation on processing.Comparing the conventional extrusion process with the extrusion-cooking (chapter 4.2.3), it wasfound that thermal treatment enhances pasta quality. Thus, the effect of pregelatinization onstarch and proteins was studied (chapter 4.2.4) and related to the quality of pasta produced atlab- (chapter 4.2.5) and pilot- (chapter 4.2.6) scale.Finally, high moisture extrusion (50%) was considered. The aptitude of air-classified pulse fractionhigh in protein was proposed as a sustainable alternative to soy protein concentrates for meatanalogues production (chapter 4.3.1).Overall, this PhD project provides evidence on how the extrusion process can be uniquely usefulfrom a pulse-product reformulation perspective and what variables are responsible fordetermining product features.
INTERPLAY BETWEEN RAW MATERIAL AND PROCESSING CONDITIONS FOR THE FORMULATION OF PULSE-BASED PRODUCTS / A. Bresciani ; tutor: A. Marti; coordinatore: D. Mora. Dipartimento di Scienze per gli Alimenti, la Nutrizione e l'Ambiente, 2022 Oct 20. 35. ciclo, Anno Accademico 2022.
STRUCTURE-BASED APPROACHES TO DATA-DRIVEN PROTEIN FOLDING, AGGREGATION, AND SELF-ASSEMBLY
Predicting protein dynamics on a molecular level is central to understanding and ultimately controlling the biomolecular machines that govern life. Despite advances in molecular dynamics and AI-based structure prediction, the accurate and efficient simulation of complex self-assembly processes - such as protein aggregation, protein folding, and the dynamics of intrinsically disordered proteins - remains a limitation of most approaches due to system size and sampling limitations. This work presents the development and application of the multi-eGO model, a data-driven, hybrid structure-based approach designed to overcome these barriers. By combining an informative prior with high-resolution structural information or lower-resolution experimental data, the multi-eGO force field learns conformational ensembles across multiple energy minima while maintaining atomistic resolution. Applications include the folding dynamics of protein G and X11-PDZ1-PDZ2, the structural ensemble of amyloid-b42 the aggregation of transthyretin peptides, and the self-assembly of ferritin complexes. Complementary to these studies, conventional molecular dynamics simulations are used to research the effect of electric fields on the dynamics of amyloid b42 fibrils, revealing the potential of electric fields to disrupt assembly and secondary nucleation. Results demonstrate that multi-eGO can reproduce equilibrium, out-of-equilibrium, and kinetic features, and integrate heterogeneous data sources, such as SAXS and PRE NMR data, to refine the model without the need for explicit training. At the same time, limitations such as finite-size effects and kinetic trapping for intermolecular processes highlight the need for further refinement.
STRUCTURE-BASED APPROACHES TO DATA-DRIVEN PROTEIN FOLDING, AGGREGATION, AND SELF-ASSEMBLY / F. Bacic Toplek ; tutor: C. Camilloni ; coordinatore: S. Ricagno. Dipartimento di Bioscienze, 2025 Nov 24. 38. ciclo
CROSSTALK BETWEEN TUMOUR VASCULATURE AND OVARIAN CANCER STEM CELLS: THE ROLE OF L1CAM.
Ovarian cancer is the most lethal gynaecologic tumour, mainly due to relapse and chemoresistance, two events fuelled by ovarian cancer stem cells (OCSC). The tumour microenvironment (TME) is thought to act as a niche for OCSC, preserving stem cells properties and sustaining tumour recurrence. The crosstalk between TME and OCSC, therefore, offers a source of new targets and mechanisms to overcome the limitations of current treatments. We identified L1CAM, an adhesion glycoprotein, expressed both in tumour endothelium and epithelial cancer cells, as a mediator of the interaction between the vascular TME (vTME) and OCSC. L1CAM expression correlates with metastatic spread and poor outcome, because of its ability to induce cancer proliferation and chemoresistance, together with the increase of vessels permeability and neo-angiogenesis. Notably, endothelial cells express a new isoform of L1CAM (L1-ΔTM), that is released as a soluble factor. We found that endothelial L1-ΔTM enhances stem-like properties in OCSC, a process mediated by IL6/JAK2/STAT3 signalling axis. Furthermore, the RNA-sequencing analysis revealed that L1-ΔTM confers a proinflammatory phenotype on OCSC, able to support cancer initiation, proliferation and progression. Moreover, I have setup a perivascular niche-on-chip confirming the capability of L1-ΔTM-expressing vessels to foster proliferation and spreading in the tumoral compartment. Our approach highlights L1-ΔTM as a druggable target in the crosstalk between vTME and OCSC. Indeed, our findings could provide the rationale for exploring novel L1CAM-based therapeutic approaches aimed at the eradication of OC through the elimination of its stem cell compartment.
CROSSTALK BETWEEN TUMOUR VASCULATURE AND OVARIAN CANCER STEM CELLS: THE ROLE OF L1CAM / M. Baronio ; supervisor: U. Cavallaro ; internal advisor: G. Scita ; phd coordinator: S. Minucci. Dipartimento di Oncologia ed Emato-Oncologia, 2024 Apr 11. 35. ciclo, Anno Accademico 2023.
EPILEPSY AND MICROBIOTA: TIME- AND DRUG-RELATED SHIFTS IN THE BACTERIAL COMMUNITY
Epilepsy is a well-known condition characterized by seizures, episodes of electricinstability provoking involuntary vigorous shaking in patients. Nowadays there aremany strategies, of which the pharmaceutical is the main one, but around one-thirdof the patients show drug resistance and, despite the different therapeuticapproaches for drug-resistant patients, many cannot reach seizure control. Indeed,it is important to look for new complementary therapeutic strategies that caninfluence the clinical picture and improve the patient’s quality of life. The connectionbetween human microbiota and neurological diseases has been thoroughly studiedover the last years. In particular, the gut-brain axis has been addressed to be a keyplayer in the development of the disease and to seizure susceptibility. Nonetheless,there is limited information so far, in literature, about the composition of theintestinal microbiota in patients with epilepsy.To check for bacterial influence on epilepsy development and treatment, during myPh.D. I’ve analyzed 3 datasets of epileptic patients: 1) a dataset of children admittedto the hospital after their first seizure and sampled 2 more times, after 4 and 12months subjected to drug monotherapy; 2) a dataset of adults that are using dailyfor more than 2 years the most common drug available with no more seizures; 3) adataset of girls affected by the Rett syndrome, a genetic disorder that comprisesimpairments in language and coordination and repetitive movements.Complications of Rett syndrome patients include seizures, and the patients take thesame drugs as the epileptic dataset. This cohort has been added to the project as anegative control, as opposed to the healthy controls enrolled. This diverse datasetset the stage for speculations on the microbiota changes and development duringepilepsy onset and constant drug therapy in terms of bacterial diversity, composition,and interaction.The bacterial DNA extracted from the fecal samples collected from the patients hasbeen sequenced through next-generation sequencing techniques on a MiSeqIllumina platform, and the resulting reads have been analyzed through abioinformatic pipeline using the QIIME software, R, and Matlab.VBiodiversity was observed to be decreased in pathological conditions and alongwith the drug assumption in all comparisons. Of particular interest is the peak ofAkkermansiaceae in the children’s drug-naive gut microbiota. The children showedan interesting intra-individual signature and a diversity separation overpharmacological treatment time, possibly related to a reduced abundance ofbeneficial groups such as Faecalibacterium. From the co-abundance networkanalysis, the opposite trend of the group of genera related to Subdoligranulum andthe ones related to Bacteroides have emerged. The adults in stable drug therapyshowed some trends similar to the Rett patients, as if, despite the different clinicalpicture, the gut bacterial community was comparable and similar in terms ofbiodiversity. The feature selection analysis performed on the drug-naive and thedrug-assuming patients confirmed the families Akkermansiaceae and Christensenellaceae to be relevant for the bacterial composition alteration during the pharmacological therapy. The separation of Gram-positive and Gram-negative bacteria showed an interesting change of the fractions with a trend in favor of theGram-negative: the different protective layers could influence or be influenced bythe drug assumption.The results shown in this thesis highlight the existence of differences in terms ofgeneral microbial diversity and taxonomy. Further studies carried out on a largernumber of patients at different times and disease development, and a metabolicanalysis implementation taking into account the nutritional intakes, could contributeto reinforcing these findings in te
EPILEPSY AND MICROBIOTA: TIME- AND DRUG-RELATED SHIFTS IN THE BACTERIAL COMMUNITY / C. Ceccarani ; tutor: E. Borghi ; co-tutore: M. Severgnini ; coordinatore: M. Samaja. Dipartimento di Scienze della Salute, 2021 Jan 21. 33. ciclo, Anno Accademico 2020. [10.13130/ceccarani-camilla_phd2021-01-21].
NOVEL APPROACHES IN COLOR ASSESSMENT: FROM POINT-WISE COLORIMETRY TO FILM RESTORATION
Colorimetry is the science which studies the quantification, reproduction and management of color from its optical, chemical, physical, but also physiological and perceptual point of view. Today, colorimetry has been standardized and color management can count on a robust workflow in point-wise conditions, thus when a color stimulus is considered under specific constraints (e.g., standard illumination and observation). As a consequence, the rigorous application of standard colorimetry is insufficient to reproduce and manage color in real conditions with non-uniform illumination and with complex spatial arrangements. This work of research is organized into three threads of work, which follow the standard pipeline of color signal evaluation: color assessment, color acquisition and processing, and quality assessment.In the first Part of this study, the assumptions and constraints of standard colorimetry are recalled, together with specific comments on their limits in non-standard and practical applications. The open problems and misuses of colorimetry discussed from the theoretical point of view are supported by preliminary tests and experiments, aiming at highlighting the limits of the application of standard colorimetry in cultural heritage field.In the second Part of this work, the standard digitization workflow is analyzed, and the issues and effects which could affect the acquisition of contrast and tones in imaging systems are discussed. In particular, the limits of hyperspectral imaging have been tested and examined, in order to assess the main source of noise in the acquisition and to evaluate the reliability of the acquired data. Subsequently, an implementation of the digitization protocol for film restoration is proposed and different methods to perform image enhancement and processing have been examined. For this aim, the family of Spatial Color Algorithms (SCAs), derived from Retinex, has been found successful for cultural heritage applications and a novel approach to film restoration is proposed. This specific family of algorithms, enhance colors according to the spatial distribution of pixel values in the scene, thus include the visual spatial mechanisms in color computation, overcoming the main constraints of standard colorimetry. Nevertheless, SCAs present high computational costs. To address this problem, in this Thesis a new speed-up algorithm is presented in order to allow the enhancement of video streams. Regarding quality assessment thread, an overview of image quality metrics is presented, and different metrics and measures have been examined and tested, to define which metrics could be the most effective and usable for cultural heritage applications. In this context, a new framework of image quality measures has been presented and applied on historical images and videos.The field of cultural heritage has been particularly suitable to underline the limits in the application of standard colorimetry, because, in this context, color is never observed as isolated phenomenon, but always inside a spatial arrangement. In this work, film digital restoration has been the main field of application and different approaches and alternative methods have been proposed to overcome the limits of standard color analysis. Anyway, the highlighted limits and solutions are applicable to film restoration, as well as to many other fields, where color sensation cannot be limited to point-wise colorimetry.
NOVEL APPROACHES IN COLOR ASSESSMENT: FROM POINT-WISE COLORIMETRY TO FILM RESTORATION / A. Plutino ; supervisor: A. Rizzi, B. R. Barricelli ; coordinator: P. Boldi. Dipartimento di Informatica Giovanni Degli Antoni, 2021 Mar 22. 33. ciclo, Anno Accademico 2020. [10.13130/plutino-alice_phd2021-03-22].
DYNAMIC INTERPLAY BETWEEN SPECTRIN, ACTIN AND PLASMA MEMBRANE DURING CELL MECHANORESPONSE
The spectrin cytoskeleton is a major component of the mammalian cell cortex. While long known and ubiquitously expressed, its dynamic behaviour and cooperation with other major components of the cell cortex is poorly understood. Here we investigated spectrin reactions upon different mechanical cues, such as cell-driven perturbations, like cell adhesion, spreading and contraction, or environmentally driven ones, like compression, stretch and osmotic changes. Upon all of these challenges we observed that spectrin meshwork spatially adapts and reorganizes under the plasma membrane together with the acto-myosin cytoskeleton. Working together to maintain cell integrity, both cytoskeletons define specific membrane territories. Actin-rich regions control protrusions, adhesions and stress fibers, whereas spectrin-rich regions concentrate in retractile zones, covering low actin density territories of the cortex. Given this interplay, we wondered if spectrin could be potentially involved in the spatial and temporal regulation of membrane trafficking. We followed spectrin, actin and clathrin dynamics through live TIRF microscopy and observed an inverted correlation between spectrin and actin densities and endocytic capacities, suggesting a spectrin contribution to clathrin-mediated endocytosis. Our results pinpoint a role for spectrin in the support of the lipid bilayer in regions where actin cytoskeleton is not established, creating a fencing mechanism for actin remodelling and cargoes internalization. All these mechanisms potentially unveil why the spectrin family of protein is evolutionary highly conserved and ubiquitously expressed in eukaryotic cells, and might explain its involvement in a broad range of pathological condition.
DYNAMIC INTERPLAY BETWEEN SPECTRIN, ACTIN AND PLASMA MEMBRANE DURING CELL MECHANORESPONSE / C. Galli ; supervisor: N. Gauthier. DIPARTIMENTO DI ONCOLOGIA ED EMATO-ONCOLOGIA, 2020 Jan 28. 31. ciclo, Anno Accademico 2019. [10.13130/galli-camilla_phd2020-01-28].
THE CHALLENGING DIAGNOSIS OF BONE DISEASE: A REAPPRAISAL OF PATHOLOGICAL BONE MARKERS BASED ON THE CAL MILANO CEMETERY SKELETAL COLLECTION
The correct identification of diseases is fundamental for the construction of the biological profile of forensic and archaeological cases. The diagnosis of bone diseases consists in the comparison of the location and morphological characteristics of bone changes with the clinical literature and previous published cases for the identification of the causative agent. While the clinical literature may not be adequate to understand the morphology and distribution of dry bone lesions, the current paleopathological literature is mostly based on archaeological skeletons, on which very little is known, and specimens from pathology museums, with extreme manifestations of diseases often unrepresentative of classic cases.This thesis aims to investigate the macroscopic diagnosis of pathological conditions based on skeletons with antemortem clinical diagnoses from a reference osteological collection: the CAL Milano Cemetery Skeletal Collection, and in particular atherosclerosis, rheumatoid arthritis, diabetes mellitus, HIV/AIDS, multiple myeloma and solid metastatic cancer. In addition to this main objective, several other research lines were explored including the histological analysis of non-skeletal calcified manifestations of diseases, the comparison between macroscopic and radiographic analysis of bone lesions and the description of bone lesions for pathological analysis on dry bones, through the examination of skeletal material from cemeterial, forensic and archaeological cases as well as samples extracted from well- preserved cadavers during autopsies.This research not only implemented the scientific literature on bone diseases but also provided specific documentation to diagnose conditions previously unexamined and raised some important issues on the diagnosis of skeletal conditions on dry bone.
THE CHALLENGING DIAGNOSIS OF BONE DISEASE: A REAPPRAISAL OF PATHOLOGICAL BONE MARKERS BASED ON THE CAL MILANO CEMETERY SKELETAL COLLECTION / L. Biehler Gomez ; tutor: C. Cattaneo. Dipartimento di Scienze Biomediche per la Salute, 2020 Feb 10. 32. ciclo, Anno Accademico 2019. [10.13130/biehler-gomez-lucie_phd2020-02-10].
UNRAVELING THE MOLECULAR PATHOGENESIS OF INEFFECTIVE ERYTHROPOIESIS IN CONGENITAL DYSERYTHROPOIETIC ANEMIA TYPE II: IN VITRO EVALUATION OF RAP-011 TREATMENT
Congenital Dyserythropoietic Anemias (CDAs) are subtypes of bone marrow failure syndromes, hallmarked by ineffective erythropoiesis. The most common form is CDA type II (CDAII), showing moderate/severe anemia, relative reticulocytopenia, jaundice, splenomegaly, and iron overload. It is inherited as an autosomal recessive disorder due to loss-of-function mutations in the SEC23B gene. Molecular pathogenesis of CDA II still has to be investigated because the described animal models did not recapitulate the clinical features observed in humans. To date, treatments for CDAII patients consist of supportive therapy, such as erythrocyte transfusions, or bone marrow transplantation or splenectomy in transfusion-dependent cases. Recently, members of TGF-β superfamily have been studied as potential regulators of erythropoiesis, especially the growth differentiation factor 11 (GDF11). Through the binding of specific receptors, GDF11 leads to an inhibited late-stage erythropoiesis. Indeed, two GDF11 inhibitors, ACE-011 and ACE-536, have been associated with an improvement of hematologic parameters. Studies with the mouse counterpart of ACE-011, RAP-011, on a mouse model of β-thalassemia showed increased differentiation of erythroid cells, improvement of the anemic condition and reduced iron overload in treated mice. The first aim of our study was the establishment of a cellular model of CDA II, that could reproduce the main defects of the disease, such as the lack of the erythroid differentiation due to the low or absent expression of SEC23B gene. For this aim, we selected the K562 cell line and, through short-hairpin RNA-based strategy, we obtained two different clones of K562 showing a stable silencing of SEC23B. Then, we decided to assess the effects of RAP-011 on this CDA II model, by investigating the pathway involved in the GDF11 signaling. This treatment simulated the ligand trap function played by RAP-011 towards GDF11. The administration of RAP-011 resulted in a reduction of SMAD2 phosphorylation induced by GDF11 and, moreover, in an increase of different erythroid differentiation markers.
UNRAVELING THE MOLECULAR PATHOGENESIS OF INEFFECTIVE ERYTHROPOIESIS IN CONGENITAL DYSERYTHROPOIETIC ANEMIA TYPE II: IN VITRO EVALUATION OF RAP-011 TREATMENT / G. De Rosa ; supervisor: A. IOLASCON ; internal advisor: B. FRANCO ; added supervisor: R. RUSSO. DIPARTIMENTO DI ONCOLOGIA ED EMATO-ONCOLOGIA, 2020 Jan 28. 31. ciclo, Anno Accademico 2019. [10.13130/de-rosa-gianluca_phd2020-01-28].
NEW TRENDS IN BREAST IMAGING FOR BREAST CANCER AND CARDIOVASCULAR RISK
Background Qualitative, subjective reading of medical images have been the backbone of image interpretation for the past century, providing useful information to the treating physician. During the past two decades, advances in medical imaging technology have offered the possibility to extract high-resolution anatomic, physiologic, functional, biochemical, and metabolic information from clinical images, all of which reflect the molecular composition of the healthy or diseased tissue of organs imaged in the human body. We are now entering the era of ―quantitative imaging‖ recently formally defined as ―the extraction of quantifiable features from medical images for the assessment of normality, or the severity, degree of change, or status of a disease, injury, or chronic condition relative to normal‖. With appropriate calibration, most of the current imaging technologies can provide quantitative information about specific properties of the tissues being imaged. Purpose This doctoral thesis aims at exploring the possible use of imaging methods such as mammography and breast magnetic resonance imaging (MRI) as imaging biomarkers, measuring functional, biochemical and metabolic characteristics of the breast through medical images. Part I. Breast arterial calcifications for cardiovascular risk Breast arterial calcifications (BAC) are easily recognizable on screening mammography and are associated with coronary artery disease. We tried to implement the estimation of BAC to be easily applicable in clinical prevention of cardiovascular disease. In particular, we evaluated the intra- and inter-observer reproducibility of i) a specifically developed semi-automatic tool and of ii) a semi-quantitative scale for BAC quantification on digital mammograms. Part II. Multiparametric breast MRI for breast cancer management Multiparametric breast MRI allows to simultaneously quantify and visualize multiple functional processes at the cellular and molecular levels to further elucidate the development and progression of breast cancer (BC) and the response to treatment. The purpose of our study was to verify the correlation between enhancement parameters derived from routine breast contrast-enhancement MRI and pathological prognostic factors in invasive BC as a condition for the use of MRI-derived imaging biomarkers in adjunct to traditional prognostic tools in clinical decision making. Part III. Artificial intelligence in Breast MRI Recent enthusiasm regarding the introduction of artificial intelligence (AI) into health care and, in particular, into radiology has increased clinicians‘ expectations and also fears regarding the possible impact of AI on their profession. The large datasets provided by and potentially extractable from breast MRI make it the right 6 stuff for fitting AI applications. This session focuses on a systematic mapping review of the literature on AI application in breast MRI published during the past decade, analysing the phenomenon in terms of spread, clinical aim, used approach, and achieved results. Conclusions Medical images represent imaging biomarkers of considerable interest in evidence- based clinical decisionmaking, for therapeutic development and treatment monitoring. Among imaging biomarkers, BAC represent the added value of an ongoing and consolidated cancer screening to act for preventing the main cause of death among women in which traditional CV risk scores do not adequately perform. Breast MRI may act as a prognostic tool to improve BC management through the extraction of a plenty of functional cancer parameters. AI might certainly implement the use of imaging data interacting with and integrating quantitative imaging for improving patient outcome and reducing several sources of bias and variance in the quantitative results obtained from clinical images. The intrinsic multiparametric nature of MRI has the greatest potential to incorporate AI applications into the so
NEW TRENDS IN BREAST IMAGING FOR BREAST CANCER AND CARDIOVASCULAR RISK / R.m. Trimboli ; avisor: F. Sardanelli. Università degli Studi di Milano, 2020 Jan 20. 32. ciclo, Anno Accademico 2019. [10.13130/trimboli-rubina-manuela_phd2020-01-20].
DISSECTING THE INTERACTION OF a-SYNUCLEIN WITH TUBULIN/MICROTUBULES
Parkinson’s disease (PD) is a neurodegenerative disorder characterized by the loss of dopaminergic neurons and the accumulation and misfolding of α-synuclein. Remarkably, PD is even the fastest-growing neurological disorder; by 2040, the number of PD patients worldwide is expected to exceed 12 million. Despite tremendous scientific research and financial investments in PD, a cure for this devastating disorder remains elusive. Therefore, a better understanding of the biological mechanisms involved in PD is crucial for the discovery of disease-modifying therapies. Here, we focus on the interaction between α-synuclein and tubulin/microtubules and its potential role in neurodegeneration. Under physiological conditions, α-synuclein is proposed to function as a so-called microtubule dynamase, by interacting with tubulin α2β2 tetramer and promoting both microtubule growth rate and catastrophe frequency in vitro. However, the interaction of PD-associated α-synuclein mutations with tubulin, and its role in PD pathogenesis has not been fully elucidated yet. In this work, we report, for the first time, that tubulin directly interacts with various PD-related α-synuclein variants and diminishes tubulin polymerization in cell-free systems. Moreover, α- synuclein aggregation is reduced in the presence of purified tubulin, suggesting that tubulin could modulate α-synuclein pathology by inhibiting its aggregation through its interaction with α-synuclein. An advanced live-cell imaging approach in a cellular model demonstrated that α- synuclein does not impact microtubule dynamics in axon-like processes. Interestingly, the expression of α-synuclein-A53T, one of the most aggregation-prone PD-related α-synuclein mutations, reduced the velocity and processivity of APP-vesicle transport along microtubules in neuronally differentiated cells. Remarkably, we disclosed that treatment with an α-synuclein aggregation inhibitor can rescue these deficits in axonal transport, suggesting that α-synuclein inclusions impact vesicle transport. In addition, we present a novel α-synuclein aggregation inhibitor that effectively inhibits α-synuclein oligomeric species in cells, which could potentially function as a disease-modifying therapeutical drug in PD. To conclude, in this study we emphasize the importance of the α-synuclein/tubulin interaction, which directly or indirectly may play a major role in neurodegeneration.
DISSECTING THE INTERACTION OF a-SYNUCLEIN WITH TUBULIN/MICROTUBULES / J.m. Kothuis ; tutor: G. CAPPELLETTI ; co-tutor: R. BRANDT. Dipartimento di Bioscienze, 2023 Sep 22. 35. ciclo, Anno Accademico 2022.
AGGREGATION MECHANISMS OF TDP-43 PROTEIN IN RESPONSE TO STRESS IN AMYOTROPHIC LATERAL SCLEROSIS AND THERAPEUTIC APPROACHES
Amyotrophic Lateral Sclerosis (ALS) is a rare neurodegenerative disorder affecting upperand/or lower motor neurons for which no effective cure is available to date. The neuropathological hallmark of the disease, present in 97% of ALS cases, consists in TDP-43 proteinopathy characterized by the accumulation of ubiquitinated and phosphorylated TAR DNA-binding protein 43 (TDP-43) in the cytoplasm, accompanied by the concomitant loss of TDP-43 splicing activity in the nucleus of affected neurons. The scarcity of experimental models of TDP-43 proteinopathy capable of recapitulating both gain and loss of TDP-43 function has hindered the discovery of novel therapeutic approaches for the treatment of ALS.Aim of this work was therefore to generate human 2D and 3D in vitro models of TDP-43proteinopathy, recapitulating TDP-43 cytoplasmic mislocalization and loss of nuclearfunction, to be used for drug screening approaches.We induced a chronic and mild oxidative insult by sodium arsenite (ARS) in humanneuroblastoma SK-N-BE cells and ALS patient-derived primary fibroblasts, inducedpluripotent stem cells (iPSCs)-motor neurons and brain organoids. As a readout of TDP-43 functionality, we tested its subcellular localization and its splicing activity towards the target genes UNC13A and POLDIP3. We demonstrated that chronic oxidative stress in these cellular models induced TDP-43 cytoplasmic mislocalization and defective TDP-43 splicing activity, thus recapitulating both TDP-43 proteinopathy features. Since we observed dysregulated gene expression of autophagy and senescence markers in ARS-treated human neuroblastoma cells, we preliminarily tested in this experimental model three drugs involved in promoting autophagy, namely rapamycin, lithium carbonate and metformin. Only rapamycin was capable of significantly rescuing ARS-induced loss of TDP-43 splicing activity on UNC13A. We therefore tested rapamycin in primary fibroblasts obtained from ALS patients with mutations in the C9orf72 gene, where we demonstrated that it reduced ARS-induced formation of phosphorylated TDP-43 (P-TDP-43) aggregates and stress granules (SGs). We also tested rapamycin in iPSCs-motor neurons from C9orf72-mutated patients, where we confirmed its efficacy in attenuating P-TDP-43 aggregates, SGs and loss of TDP-43 splicing activity on UNC13A induced by ARS treatment. We then employed a more complex 3D neuroglial model, namely iPSC-brain organoids, obtained from C9orf72 ALS patients and exposed to chronic ARS, where we observed that rapamycin rescued loss of TDP-43 splicing activity on UNC13A gene.In conclusion, we generated and characterized different human cell models of TDP-43proteinopathy in which rapamycin was able to rescue chronic oxidative stress-induced TDP-43 cytoplasmic mislocalization and defects in its splicing activity. We therefore propose that human SK-N-BE cells and ALS patient-derived 2D and 3D iPSC models chronically treated with ARS can be exploited as in vitro models to be used for future drug screening approaches.
AGGREGATION MECHANISMS OF TDP-43 PROTEIN IN RESPONSE TO STRESS IN AMYOTROPHIC LATERAL SCLEROSIS AND THERAPEUTIC APPROACHES / V. Casiraghi ; tutor: A. Ratti ; director: N. Landsberger. Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, 2024 May 09. 36. ciclo, Anno Accademico 2023.
COFFEE AND TEA CONSUMPTION AND GASTRIC CANCER: A POOLED ANALYSIS IN AN INTERNATIONAL CONSORTIUM OF EPIDEMIOLOGICAL STUDIES.
The main focus of my doctoral research was to examine the effect of coffee and tea drinking on gastric (stomach) cancer risk. Coffee and tea are among the most popular drinks worldwide after water, and their consumption is expected to rise by an average of 17% over the next several years. The World Cancer Research Fund and the American Institute for Cancer Research (WCRF/AICR) concluded in their most recent report (2018) that there is insufficient data to draw any conclusions about the relationship between coffee and tea consumption and gastric cancer. To estimate their relationship with gastric cancer risk, individual-participant pooled analyses in a unique global consortium of epidemiological studies on gastric cancer - the Stomach cancer Pooling (StoP) project- were carried out. The StoP project included thirty case-control (CC) studies and five nested case-control within the cohort (NCC) studies, from seventeen different countries worldwide. For each study, coffee and tea consumption were assessed using self- or interviewer-administered food frequency questionnaires (FFQ) prior to the diagnosis of gastric cancer cases or prior to the onset of disease, hospital admission for hospital-based controls, or recruitment for population-based controls. Data on coffee intake were available from eighteen studies, which included 8,198 gastric cancer cases and 21,419 controls, while on tea twenty-two studies totaled 9,438 gastric cancer cases and 20,451 controls. Coffee and tea consumption was measured by the number of cups, times, or frequency of consumption reported in each study and further expressed in the standard unit of cups per day. Coffee was considered either caffeinated coffee or decaffeinated coffee separately, as well as their combined intake. Various types of tea reported were calculated as total tea intake. In addition, the temperature and strength at which tea was consumed were assessed. Logistic generalized linear mixed-effects models including terms Dissertation Advisor: Professor Carlo La Vecchia Georgia Martimianakifor sex, age, study design, Helicobacter pylori (H. pylori) infection, and several other main recognized risk factors for gastric cancer were used to estimate the associations between gastric cancer and coffee and tea consumption, through both two- and one-stage modeling analyses. Subgroup analyses across strata of several factors and dose-response relationships were also carried out. Multinomial mixed-effects models were also applied to estimate the ORs and corresponding 95% CIs of gastric cancer by the anatomical site (cardia and non-cardia) and histological type (intestinal, diffuse, and mixed/unspecified by Lauren categorization) for regular tea drinkers and high coffee drinkers. Heterogeneity between strata was evaluated by Cohran’s Q test and by the I2 test between studies. The results showed a slightly inverse association (OR: 0.92, 95% CI: 0.82–1.05) between tea drinking and gastric cancer but no relevant association (OR: 1.03, 95% CI: 0.94-1.13) between coffee consumption and gastric cancer. Regular tea drinkers had a lower gastric cardia cancer risk (OR: 0.64, 95% CI: 0.49-0.84), than heavy coffee drinkers, who were positively associated (OR:1.61, 95% CI: 1.27–2.05) with gastric cardia cancer. Contrary to tea intake, which was more strongly linked to lower risk in Asian countries (OR: 0.67, 95% CI: 0.49-0.91, in studies from China and Japan), there was no evidence of regional differences in the effect of coffee consumption on the risk of gastric cancer. Consuming hot or very hot tea did not increase the risk of gastric cancer, drinking warm or cold tea was related to a lower risk.Last but not least, I also discuss additional research projects I have undertaken through my doctoral studies. These include other dietary analyses within the Stomach Cancer Pooling project co
COFFEE AND TEA CONSUMPTION AND GASTRIC CANCER: A POOLED ANALYSIS IN AN INTERNATIONAL CONSORTIUM OF EPIDEMIOLOGICAL STUDIES / G. Martimianaki ; tutor: LA VECCHIA, CARLO VITANTONIO BATTISTA. Dipartimento di Scienze Cliniche e di Comunità, 2023 Jan 17. 35. ciclo
A POWER INDEX BASED FRAMEWORKFOR FEATURE SELECTION PROBLEMS
One of the most challenging tasks in the Machine Learning context is the feature selection. It consists in selecting the best set of features to use in the training and prediction processes. There are several benefits from pruning the set of actually operational features: the consequent reduction of the computation time, often a better quality of the prediction, the possibility to use less data to create a good predictor.In its most common form, the problem is called single-view feature selection problem, to distinguish it from the feature selection task in Multi-view learning. In the latter, each view corresponds to a set of features and one would like to enact feature selection on each view, subject to some global constraints.A related problem in the context of Multi-View Learning, is Feature Partitioning: it consists in splitting the set of features of a single large view into two or more views so that it becomes possible to create a good predictor based on each view. In this case, the best features must be distributed between the views, each view should contain synergistic features, while features that interfere disruptively must be placed in different views.In the semi-supervised multi-view task known as Co-training, one requires also that each predictor trained on an individual view is able to teach something to the other views: in classification tasks for instance, one view should learn to classify unlabelled examples based on the guess provided by the other views.There are several ways to address these problems. A set of techniques is inspired by Coalitional Game Theory. Such theory defines several useful concepts, among which two are of high practical importance: the concept of power index and the concept of interaction index. When used in the context of feature selection, they take the following meaning: the power index is a (context-dependent) synthesis measure of the prediction’s capability of a feature, the interaction index is a (context-dependent) synthesis measure of the interaction (constructive/disruptive interference) between two features:it can be used to quantify how the collaboration between two features enhances their prediction capabilities. An important point is that the powerindex of a feature is different from the predicting power of the feature in isolation: it takes into account, by a suitable averaging, the context, i.e. the fact that the feature is acting, together with other features, to train a model.Similarly, the interaction index between two features takes into account the context, by suitably averaging the interaction with all the other features.In this work we address both the single-view and the multi-view problems as follows.The single-view feature selection problem, is formalized as the problem of maximization of a pseudo-boolean function, i.e. a real valued set function (that maps sets of features into a performance metric). Since one has to enact a search over (a considerable portion of) the Boolean lattice (without any special guarantees, except, perhaps, positivity) the problem is in general NP-hard. We address the problem producing candidate maximum coalitions through the selection of the subset of features characterized by the highestpower indices and using the coalition to approximate the actual maximum.Although the exact computation of the power indices is an exponential task, the estimates of the power indices for the purposes of the present problem can be achieved in polynomial time.The multi-view feature selection problem is formalized as the generalization of the above set-up to the case of multi-variable pseudo-boolean functions.The multi-view splitting problem is formalized instead as the problem of maximization of a real function defined over the partition lattice. Also this problem is typically NP-hard. However, candidate solutions can be found by suitably partitioning the top power-index features
"A POWER INDEX BASED FRAMEWORKFOR FEATURE SELECTION PROBLEMS / C. Mio ; tutor: G. Gianini; supervisore: E. Damiani ; coordinatore: P. Boldi. DIPARTIMENTO DI INFORMATICA ""Giovanni Degli Antoni"", 2020 Jan 31. 32. ciclo, Anno Accademico 2019. [10.13130/mio-corrado_phd2020-01-31]."
MOLECULAR PATHWAYS ANALYSIS THROUGH MULTI-OMICS APPROACHES TO STUDY THE BIOLOGICAL BASIS OF ANOREXIA NERVOSA
Anorexia nervosa is a complex and multifactorial disorder with a high genetic component and severe metabolic consequences. Despite its high heritability, the molecular mechanisms underlying AN remain unclear, and current treatment strategies are often ineffective. Recent advancements in genomics and metabolomics offer promising tools for understanding the genetic and biochemical pathways involved in AN. This study aimed to investigate the genetic and metabolomic basis of AN using next-generation sequencing, polygenic risk scoring, and hair metabolomic profiling, with the ultimate goal of improving diagnosis and treatment strategies.This study analyzed three distinct case studies involving AN patients. The first case study involved sequencing the DNA of 68 AN patients using a panel of 162 genes to identify rare variants associated with the disorder. The second case study expanded the cohort to 135 AN patients and included the calculation of PRS using a newly designed panel of 163 genes and 730 single nucleotide polymorphisms. The third case study focused on metabolomic analysis of hair samples from 25 AN patients and 25 healthy controls, with an emphasis on amino acid and carnitine profiles. NGS, statistical analysis, and machine learning algorithms were employed to analyze genetic variants and metabolomic profiles.In the first case study, rare variants in key genes were identified, suggesting a potential monogenic form of AN. The second case study revealed a significant genetic contribution to AN risk, with the PRS explaining 21.3% of the variance in the phenotype. Variants in genes related to dopamine signaling, skeletal muscle function, and appetite regulation were identified, highlighting diverse molecular mechanisms contributing to AN. The third case study demonstrated significant metabolic alterations in AN patients, particularly deficiencies in branched-chain amino acids and essential amino acids. These findings suggest a biochemical basis for appetite dysregulation observed in AN patients. Given the emerging evidence from genetic studies, genetic testing in AN is becoming a clinical reality, with identified candidate genes such as NNAT, EPHX2, LEP, and MC4R offering the potential to tailor treatments, predict disease progression, and assess recurrence risk within families. Moreover, genetic testing is crucial for distinguishing between true AN and syndromic forms that mimic AN but have distinct etiologies, enabling more accurate diagnoses and avoiding mismanagement.This study provides insights into the genetic and metabolic foundations of AN. The integration of genomic and metabolomic data highlights the potential for personalized medicine approaches in AN diagnosis and treatment. Genetic testing, particularly for rare variants, could play a key role in early diagnosis and risk assessment, while targeted nutritional interventions may address the metabolic imbalances associated with the disorder. This study investigates a large Italian cohort of 228 anorexia nervosa (AN) patients to validate genetic associations with AN. We developed a genetic test targeting syndromic forms that mimic AN phenotypically but have distinct underlying etiologies. Our analysis identified key variants in genes such as NNAT, EPHX2, and ESR2, highlighting their role in AN pathogenesis. Syndromic forms related to genes like ARG1 and ALPL were also explored. This genetic test represents a crucial tool for differential diagnosis, improving clinical outcomes for patients with AN and its syndromic forms. Future research should focus on expanding these findings to larger cohorts and further exploring the interaction between genetic predisposition and environmental factors in AN.
MOLECULAR PATHWAYS ANALYSIS THROUGH MULTI-OMICS APPROACHES TO STUDY THE BIOLOGICAL BASIS OF ANOREXIA NERVOSA / K. Donato ; tutor accademico: R. C. Paroni ; tutor industriale: M. Bertelli ; coordinatore: C. Sforza. - Università degli Studi di Milano. Dipartimento di Scienze della Salute, 2025 Jan 24. 37. ciclo
A RANDOMIZED STUDY OF THE GLUNOVO REAL-TIME CGM EFFECTIVENESS ON METABOLIC CONTROL IN PATIENTS WITH T2D
Objective: This study aimed to evaluate the effectiveness of the real-time continuous glucose monitoring (RT-CGM) system, Glunovo, in improving glycemic control and patient outcomes in individuals with poorly controlled type 2 diabetes (T2D) compared to traditional self-monitoring of blood glucose (SMBG).Research Design and Methods: This prospective, open-label, randomized controlled trial included 178 patients with T2D from the Fatebenefratelli-Sacco Hospital in Milan. Participants were randomized into 2 groups: 89 patients received the Glunovo RT-CGM (case group), while 89 continued SMBG with standard glucometers (control group). The primary outcome was the change in HbA1c levels after 6 months. Secondary outcomes included time in range (TIR), time above range (TAR), time below range (TBR), mean glucose levels, Glucose Management Indicator GMI and patient satisfaction and well-being assessed by a numeric scale and the WHO-5 Well-Being Scale.Results: After 6 months, the RT-CGM group showed a significant reduction in HbA1c levels (Δ -1.4%, p<0.001), compared to the control group (Δ -0.6% p<0.001). TIR significantly increased in the RT-CGM group (Δ +18.4%, p<0.001). TAR significantly decreased in the RT-CGM group (Δ -17.2%, p<0.001). There was no significant change in TBR in RT-CMG group. The GMI in the RT-CGM group dropped by 1.4% (p<0.001). Satisfaction with the RT-CGM significantly increased (p<0.001), but no significant improvements were observed in the well-being (WHO-5 index).Conclusions: The use of the Glunovo RT-CGM system significantly improved glycemic control and patient satisfaction compared to SMBG. These findings suggest that Glunovo RT-CGM is an effective tool for managing poorly controlled T2D, providing a valuable option for enhancing both glucose regulation and patient self-management through improved dietary awareness and optimized therapy adherence.
A RANDOMIZED STUDY OF THE GLUNOVO REAL-TIME CGM EFFECTIVENESS ON METABOLIC CONTROL IN PATIENTS WITH T2D / E. Lazzaroni ; tutor: P. Fiorina, F. D'Addio ; coordinator: F. Cheli. Dipartimento di Scienze Biomediche e Cliniche, 2025 Jun 30. 37. ciclo, Anno Accademico 2023/2024.
SOCIAL AND REPUTATIONAL DRIVERS OF SUNK-COST SENSITIVITY IN INDIVIDUAL AND COLLECTIVE DECISION-MAKING
Why do people stick with failing plans after investing resources? This dissertation argues that sunk-cost behavior is shaped more by social pressures than by cognitive bias. Across three preregistered, “waste-free” studies, I show that people persist to protect reputation and relationships—especially when others have invested or when commitment signals reliability. When social audiences or obligations disappear, escalation fades. Additional chapters show that voters reward decision-makers who stay committed and develop a framework explaining both cognitive and social costs of changing course. Together, the work explains when sunk costs bind—and when they don’t.
SOCIAL AND REPUTATIONAL DRIVERS OF SUNK-COST SENSITIVITY IN INDIVIDUAL AND COLLECTIVE DECISION-MAKING / M. Mattei ; tutor: J. Michael ;secondo tutor: D. Buehlersupervisori periodo visiting: N. Sebanz; G. Knoblich. Dipartimento di Filosofia Piero Martinetti, 2025 Jan. 38. ciclo, Anno Accademico 2024/2025.
REPRODUCTION AND PERINATOLOGY IN COMPANION ANIMALS INVESTIGATED BY USE OF NONINVASIVE MATRICES
Reproduction in companion animals is a topic of growing interest for scientific research, but knowledge about some aspects is still lacking. This is likely due to the need of collecting a large number of serial samples in order to perform longitudinal studies of the longer-lasting reproductive phases, which is now incompatible with the current concept of animal welfare, according to which any distress must be avoided, especially where pregnant and neonatal subjects are concerned. For these reasons, complying with the need of respecting animal welfare and aiming to study reproductive phases that are still incompletely explored in companion animals, the project lines of the present PhD thesis were focused on three crucial phases of reproduction: pregnancy and post-partum, perinatology and puberty. This was achieved by using matrices, like coat and the claws, and, to a lesser extent, fetal fluids obtained by noninvasive sampling. The sudden Covid-19 pandemic and the consequent lockdown rules have partially impaired the sample collection and laboratory analyses, but some interesting and satisfactory data were provided nonetheless. In the pregnancy and post-partum study, the results showed changes in the concentrations of cortisol (C) in coat from mating to 60 days post- partum, in line with what was reported for cats, suggesting that canine maternity could be considered as a challenge for the bitches, and dehydroepiandrosterone-sulfate DHEA(S) was also analyzed. About perinatology, the claws concentrations of C, DHEA(S), E2 and T tended to decrease from birth until 60 days of age, adding precious information about the perinatal physiology. In fetal fluids, leptin amniotic concentrations were higher in smaller-seized breeds, highlighting the role of breed body-size in affecting the intrauterine fetal metabolism. In the puberty study, in cats 17β-estradiol (E2) and testosterone (T) concentrations were assessed in coat and dewclaws. The concentrations of T in coat significantly differed between pubertal males and females and between gonadectomized males and females. The concentrations of T in coat were higher in pubertal than prepubertal male cats. In dogs, the analyses of the concentrations of C, DHEA(S), T and E2 in coat showed lower DHEA(S) concentration in pubertal than prepubertal and gonadectomized female dogs. In males, T concentrations in coat were higher in pubertal than prepubertal dogs, and C concentrations in coat were higher in pubertal than prepubertal and gonadectomized dogs, suggesting that pubertal status leads to a higher activation of the HPA axis in male dogs. The results shown in the present thesis evidenced the usefulness of these matrices for longitudinal and long-term hormonal studies of reproduction in dogs and cats, providing useful data about crucial reproductive phases and new interesting insights. Further investigations are needed to better understand some pending questions about the use of these matrices.
REPRODUCTION AND PERINATOLOGY IN COMPANION ANIMALS INVESTIGATED BY USE OF NONINVASIVE MATRICES / J. Fusi ; tutor: M. C. Veronesi ; PhD Program Coordinator: V. Grieco. Dipartimento di Medicina Veterinaria, 2021 Jan 28. 33. ciclo, Anno Accademico 2020.
EPIDEMIOLOGY AND MOLECULAR CHARACTERIZATION OF SELECTED PROTOZOA IN DOMESTIC RUMINANTS
Apicomplexa parasites are single-celled, obligate intracellular cyst-forming protozoa, infecting humans and animals, that pose major threats to world health and global economy. Among the most relevant species for farm animals, Besnoitia besnoiti, Neospora caninum and Toxoplasma gondii are parasites of medical (T. gondii) and veterinary (T. gondii, N. caninum, B. besnoiti) importance in domestic ruminants.Bovine besnoitiosis, caused by Besnoitia besnoiti, is a chronic and debilitating parasitic disease of cattle, characterized by both cutaneous and systemic manifestations, compromising animal welfare and responsible for economic losses on affected farms. In Europe, including Italy, bovine besnoitiosis is an emerging or re-emerging disease, with an increasing geographical distribution and the number of cases of infection. Neospora caninum, a coccidian protozoan, represents an important cause of bovine abortion. It was suggested that the parasite may have adverse effects on fertility and milk production, but only few and contrasting data are available to date. Besides, while only a single genotype of N. caninum exists worldwide, available parasite strains show considerable variation in vitro and in vivo, including different virulence in cattle. Microsatellite markers allow to fingerprint N. caninum isolates or DNAs and undertake population studies. Toxoplasmosis represents an important public health issue, with the consumption of raw or undercooked meat being a major way of human infection. The role of beef in the transmission of the parasite to humans is questioned due to lower quantity of tissue cysts compared with other meat-producing species. However, the habit of consuming raw beef is regionally diffused, and the risk posed by Toxoplasma gondii infection in cattle should not be overlooked. The aim of my doctoral thesis project was to investigate on the epidemiology and molecular characterization of selected protozoa parasites of medical and veterinary importance in domestic ruminants, i.e., B. besnoiti, N. caninum and T. gondii in cattle. A multidisciplinary approach based on clinical features, laboratory tests including serological and molecular techniques, was applied throughout the research project, to achieve a multi-level comprehension of the epidemiology of these parasite infections. Three main research lines were developed: Research Line 1: Exploring bovine besnoitiosis: a multi method approach. A case of bovine besnoitiosis in a dairy farm housing 217 cattle in Italy was reported. A serological screening was performed on the whole herd using the recommended approach of ELISA and confirmatory Western Blot. Seropositive animals were clinically examined to reveal symptoms and lesions of besnoitiosis. Risk factors and the effects of the parasite infection on reproductive and productive performances were evaluated. Histopathology and molecular analyses on tissues from a slaughtered cow affected by the chronic phase of the disease were carried out. An overall seroprevalence of 23.5%, which increased up to 43.5% considering only cows, was recorded. Clinical examination of 33 of the seropositive cows evidenced the presence of tissue cysts in at least one of the typical localizations (sclera, vulva, or skin) in 25 animals. Statistical analysis did not evidence any significative impact of the parasite infection on herd efficiency; however, a decrease of productive parameters was recorded in cows showing cutaneous cysts. Concerning the chronically affected cow, histopathology revealed B. besnoiti tissue cysts in the skin of the neck, rump, hind legs, eyelid and vulva, in the muzzle, in mucosal membranes of the upper respiratory tract, and in the lungs. Parasite DNA was detected also in masseter muscles, tonsils, mediastinal lymph nodes, liver, cardiac muscle, aorta wall, ovaries, uterus, and vulva.Moreover, alterations of laboratory parameters, i.e., hematological and biochemical parameters, enzyme activ
EPIDEMIOLOGY AND MOLECULAR CHARACTERIZATION OF SELECTED PROTOZOA IN DOMESTIC RUMINANTS / L. Villa ; tutor: M. T. Manfredi ; coordinatore: V. Grieco. Dipartimento di Medicina Veterinaria, 2021 Jan 28. 33. ciclo, Anno Accademico 2020.
DEPICTING MOTION IN STATIC IMAGES. A PHILOSOPHICAL INQUIRY
In my thesis, I deal with the rarely discussed issue of whether motion can be depicted in static images and I provide a novel, and composite, view on the matter. Part I focuses on whether various pictures usually considered to be effective in representing motion are, actually, proper depictions of motion. Part II takes up two issues concerning distinctive aspects of depiction as a peculiar kind of representation – pictorial realism and inflection - and considers them in light of motion depiction and vice versa.In the first chapter, I focus on pictures that depict objects caught in the middle of dynamic actions, and, building on psychological studies on implied motion, I argue they can properly depict motion. In fact, I maintain, motion is part of the perceptual content of our experience of such pictures – and, hence, of their figurative content - even in the absence of motion-like phenomenology. In the second chapter, I deal with various devices exploited in different pictorial styles – such as comics’ motion marks or multiple images - and argue that some of them are actual depictions of aspects of our experiences of motion since they elicit recognition responses for actual optical effects we can experience in real-life situations when tracking fast-moving objects. In the third chapter, I discuss long exposure photographs resulting in streaky images, chronophotographs, and futurists’ paintings and argue we do not interpret them pictorially, but that to understand the temporal content of such images we pretend: we play games of make-believe using what we properly see in the picture – which is what is properly depicted – as a prop. In the fourth chapter, I consider two cases of optical illusions of movement - Op art’s scintillating effects and peripheral drift illusions - and conclude that one of them, the latter, is involved in the depiction of movement. All the accounts developed in the first part have profound consequences for thinking about the nature of pictures and pictorial experience – in particular, they often act as stress tests for different theories of depiction.In the first chapter of the second part, I then argue that various instances of both depicted and merely represented motion analyzed in the previous part enlighten different aspects of the concepts of pictorial realism (and unrealism); furthermore, I show how, vice versa, existing accounts of realism make the limits of motion depiction manifest. Finally, I argue there is a sense in which we can say that motion – or, better, the sense(s) of motion - can be inflected in static pictures: this analysis, on the one hand, expands the notion of inflection and its reach, and, on the other, gives a framework for analyzing the relationships between what has often been acknowledged to be a central aspect of our aesthetic appreciation of pictures – their dynamism – and pictorial experience – intended as seeing-in.
DEPICTING MOTION IN STATIC IMAGES. A PHILOSOPHICAL INQUIRY / L. Marchetti ; tutor: P. Spinicci, C. Calabi ; coordinatore: A. Pinotti. Università degli Studi di Milano, 2022 Oct 24. 34. ciclo, Anno Accademico 2021.
GIUSTIZIA PENALE E DECISIONI ALGORITMICHE PROSPETTIVE DEI RISK ASSESSMENT TOOLS NORDAMERICANI TRA PROGRESSO TECNOLOGICO, INTELLIGENZA ARTIFICIALE E GIUSTO PROCESSO
The main goal of this research is to assess the compatibility between the use of “predictive tools” in the criminal process and the protection of due process principles and values. In particular, the research primarily focuses on algorithmic risk assessment tools that assist judges in assessing the defendant’s dangerousness and likelihood of recidivism. The aim is to take into consideration the advantages of applying these algorithmic tools in some key junctures of criminal proceedings, both in terms of efficiency and objectivity and uniformity of decisions. At the same time, it is of crucial importance to carefully consider the risks associated with the use of algorithmic systems with regard to the fundamental tenets of due process and fair trial rights (and in particular, right to defense counsel, the protection against self- incrimination, the presumption of innocence, and judicial impartiality).The research will be carried out through a comparison between the US and Italian legal systems. In the United States, the use of AI, big data, and machine learning tools in legal practice has now become a reality, and in recent years we have been witnessing a noticeable growth in the use of algorithmic risk assessment tools at all stages of the criminal process. In the Old Continent, on the other hand, the use of algorithmic tools has not yet reached the level of diffusion comparable to U.S. jurisdictions, and the first official reports and policy documents on the development of algorithmic and AI systems have only recently been published. This research aims to assess whether similar tools, if undestood and corrected, could be implemented also in the Italian criminal justice, and what “natural defenses” the Italian system has for protecting the fundamental rights of individuals involved in the assessment of these tools.
"GIUSTIZIA PENALE E DECISIONI ALGORITMICHE: PROSPETTIVE DEI RISK ASSESSMENT TOOLS NORDAMERICANI TRA PROGRESSO TECNOLOGICO, INTELLIGENZA ARTIFICIALE E GIUSTO PROCESSO / A. Di Domenico ; tutor: L. Camaldo ; co-tutor: N. Galantini ; coordinatrice: F. Poggi. - Dipartimento di Scienze Giuridiche ""Cesare Beccaria"". Dipartimento di Scienze Giuridiche Cesare Beccaria, 2024 Jul 12. 36. ciclo, Anno Accademico 2023/2024."
ELUCIDATING THE IMPACT OF ASCITES ON HIGH GRADE SEROUS OVARIAN CANCER THROUGH MULTIPLE IN VITRO PATIENT-SPECIFIC MODELS
High-grade serous ovarian cancer (HGSOC) is a highly heterogeneous and aggressive malignancy characterized by marked inter-/intra-tumor variability and a dynamic tumor microenvironment (TME). The TME, and particularly ascitic fluid (AF), plays a critical role in OC progression by influencing tumor and stromal cell behavior, promoting metastasis and driving therapy resistance. Current in vitro models often fail to replicate the complexity of the TME, being limited in their ability to fully capture HGSOC biology. Here, we explored the impact of AF supplementation on multiple patient-derived in vitro models, specifically 2D cells, 3D stem-enriched spheroids and polyclonal organoids, to develop preclinical models able to capture and faithfully reproduce the heterogeneity and complexity of this tumor and its TME.We could show that AF supplementation improves culturing conditions by increasing cell proliferation, sphere formation and propagation, while also preserving the original metabolic profiles of patient-derived samples. In parallel, 2D cells and 3D spheroids revealed a model-specific effect of ascites, with a downregulation of genes involved in the biosynthesis of cholesterol and in inflammatory response in 2D cells, and an upregulation of proliferation and glycolysis in 3D spheres. Additionally, we identified a common effect of AF on both models, showing AF-dependent genes and pathways related to cell cycle regulation and cellular proliferation. Among them, CDKN2B was notably downregulated in the presence of AF, highlighting a novel regulatory mechanism directly driven by the ascitic TME in HGSOC.Integrative analyses revealed that the in vitro models were mostly enriched for tumor stroma cellular subpopulations, warranting a perspective shift from a tumor epithelial-centric view to a systematic, and now finally experimentally tractable, appraisal of the TME, particularly cancer-associated fibroblasts (CAFs), paving the way to a refined understanding of tumor progression in light of tumor-TME cross-interaction. In this context, the study underscores the critical role of AF in shaping the interactions between tumor and non-tumor cells and the surrounding microenvironment, highlighting the importance of TME for a comprehensive understanding of HGSOC biology. The integration of AF into in vitro systems represents a significant step forward in creating more defined models for studying stroma and/or tumor interactions, allowing in the future to position these models within the broader landscape of HGSOC biology and potentially guide more refined therapeutic strategies.
ELUCIDATING THE IMPACT OF ASCITES ON HIGH GRADE SEROUS OVARIAN CANCER THROUGH MULTIPLE IN VITRO PATIENT-SPECIFIC MODELS / B. Barzaghi ; tutor: G. Testa; co-supervisor: P. Lo Riso ; coordinator: D. Pasini. Dipartimento di Oncologia ed Emato-Oncologia, 2025. 36. ciclo, Anno Accademico 2023/2024.
HABILIDAD ARGUMENTATIVA EN HISPANOHABLANTES CON AFASIA PROGRESIVA PRIMARIA: UNA COMPARACIÓN TRANSCULTURAL SEGÚN EL NIVEL DE ESCOLARIZACIÓN Y EL REPERTORIO LINGÜÍSTICO
Primary Progressive Aphasia (PPA) is a neurological syndrome related to several neurodegenerative diseases and characterised by a progressive loss of speech. Within the discourse studies on APP, there is a lack of focus on one of the most complex discursive skills, i.e. argumentative skills. For this reason, the aim of of this thesis is to detect and quantitatively analyse the argumentative ability of a group of patients with PPA compared to a control group, according to two parameters of comparison: the level of schooling and their background linguistic knowledge. During a research stay at the San Carlos Clinical Hospital in Madrid, 30 PPA patients and their controls were given an argumentative test, created ad-hoc. The test, which was audio-visually recorded to capture gestures and proxemics, is structured in three parts, each corresponding to a different argumentative stimulus (textual, graphic, conversational). By means of a text, images and a debate, it was possible to collect data from three different perspectives of argumentative production or comprehension. After the test, two questionnaires were administered in order to collect information on levels of schooling (My linguistic biography) and the languages spoken before (Levels of language proficiency). After having transcribed the tests with the audio transcription software WhisperAI and merging the transcripts into a corpus of PPA argumentative discourse, it was possible to insert the answers to the textual analysis questions and the interpretations of the images into a matrix of SPSS programme, in order to make a statistical analysis of the data. At the same time, it was possible to create a PPA argumentative profile model by classifying the arguments used by patients (according to the Wagemans’ classification, 2020) and calculating the frequency of use of the argumentative markers. By means of the Chi-square test in SPSS, it was possible to see the percentage of success of the PPA patients in the textual analysis and in the interpretation of the images, with respect to the controls and, consequently, the survival or not of certain persuasive skills. The influence of schooling level and linguistic background wascalculated through a comparison of means and, in the case of schooling, also through Spearman's correlation index. The main finding of the study is that patients with APP show impaired argumentative ability and no significant correlations are observed with school level or linguistic reserve. The finding that a subgroup of APP patients show preserved argumentative ability opens the door for future studies to better understand the reasons underlying such preservation in APP. In addition, there are qualitative differences in argumentative procedures that could be addressed in future research.
HABILIDAD ARGUMENTATIVA EN HISPANOHABLANTES CON AFASIA PROGRESIVA PRIMARIA: UNA COMPARACIÓN TRANSCULTURAL SEGÚN EL NIVEL DE ESCOLARIZACIÓN Y EL REPERTORIO LINGÜÍSTICO / A. Donati ; tutor: M. V. Calvi ; co-tutor: O. Ivanova, J. Matías-Guiu. Dipartimento di Lingue, Letterature, Culture e Mediazioni, 2025 Jan 27. 37. ciclo, Anno Accademico 2023/2024.
ACTIONS DERIVING DIRECTLY FROM INSOLVENCY PROCEEDINGS AND CLOSELY LINKED WITH THEM UNDER REGULATION EU 848/2015 ON INSOLVENCY PROCEEDINGS
Historically, the topic of individual actions directly deriving from and closely linked with insolvency proceedings (hereinafter referred to as “Annex Actions”) has been addressed in the European scenario with reference to the scope ratione materiae of the Brussels Convention dated 1968 on the jurisdiction, recognition and enforcement of judgments in civil and commercial matters (the “Brussels Convention”). In the absence (at that time) of Community rules on cross-border insolvencies, in the Gourdain judgment the ECJ interpreted the notion of Annex Actions under the prism of the exception relating to “bankruptcy, proceedings relating to the winding-up of insolvent companies or other legal persons, judicial arrangements, compositions and analogous proceedings”, set forth by Article 1(2)(2) Brussels Convention. It was on that occasion that the Court held that actions which are related to insolvency proceedings are excluded from the scope of the Brussels Convention (now the Brussels 1a Regulation), with the proviso that they derive directly from insolvency proceedings and are closely linked with them (thus coining what has been called, over the years, the “Gourdain Formula”). The interpretative uncertainties arising from such an ambiguous wording - at the time referring only to the scope of application of the Brussels Convention - were not removed (and possibly were even exacerbated) by the introduction of the European regime on insolvency proceedings. Indeed, the Regulation (EC) 1346/2000 (the “Insolvency Regulation”), on the one hand, revealed a serious regulatory gap, since it did not provide for a rule on the jurisdiction of Annex Actions and, on the other hand, even where it mentioned them (for purposes other than jurisdiction), it laconically restated the Gourdain Formula, with no further clarifications.The nebulous legislative framework described above has been partly clarified by Regulation (EU) 2015/848 (the “Recast Regulation”), which has remedied the aforementioned omission, expressly providing for the (halved) European vis attractiva concursus. According to that principle, the courts of the Member State in the territory of which insolvency proceedings are opened, are vested with the jurisdiction to hear and determine Annex Actions. The impact of the reform over the “dynamic” profile of the vis attractiva concursus must be positively assessed since it has dispelled many of the doubts concerning the allocation of jurisdiction on Annex Actions. Yet, the problem of the autonomous definition of Annex Actions remains partially unsolved, because also under the Recast Regulation, the contours of that concept continue to be defined by the vague Gourdain Formula.Only partial indications can be drawn from Recital 35 Recast Regulation and, above all, from the extensive case-law of the ECJ. The latter, however, allows only to some extent to draw a systematic notion of Annex Actions, as it substantially reveals a wavering orientation, which does not permit to trace (yet) a general criterion, certain and shared on this point. It is suggested that, for the purposes of the European legislation, Annex Actions are those actions underpinning a right or obligation which stems from the opening of insolvency proceedings, whose DNA, we might say, is ontologically linked to insolvency proceedings. They would count a very small number of actions such as (predictably) avoidance actions, liability actions against the trustee and other bodies of the procedure, and actions arising from the termination of contracts exercised by the trustee by virtue of the express powers conferred upon him by insolvency law. On the contrary, all other actions in respect of which the procedure is a neutral (legal) event should not be characterised as Annex Actions (e.g. actions that have only an occasional link with insolvency proceedings and those that existed in the legal sphere of the insolvent debto
ACTIONS DERIVING DIRECTLY FROM INSOLVENCY PROCEEDINGS AND CLOSELY LINKED WITH THEM UNDER REGULATION EU 848/2015 ON INSOLVENCY PROCEEDINGS / C.t.m. Lunetti ; tutor: . Bariatti ; co-tutor: E. Pataut ; coordinator: D. U. Galetta. Università degli Studi di Milano, 2020 Feb 14. 32. ciclo, Anno Accademico 2019. [10.13130/lunetti-chiara-teresa-maria_phd2020-02-14].
ROLE OF NATURAL KILLER CELLS IN THE PATHOGENESIS OF PRIMARY BILIARY CHOLANGITIS
"Background: Primary biliary cholangitis (PBC) is the most prevalent autoimmune liver disease and involves the selective destruction of small intrahepatic biliary epithelial cells (BECs), potentially leading to biliary cirrhosis and liver transplantation, if left untreated. As of now, anticholestatic therapy, in particular Ursodeoxycholic acid (UDCA), stands as the primary treatment approach for PBC, serving as the established first-line therapy irrespective of the disease stage. Unfortunately, up to 40% of PBC patients experience treatment failure with UDCA, facing an elevated risk of disease progression, while the survival rate among biochemical ""responders"" aligns with that of the general population. Therefore, there is urgent need to better characterize the features of PBC patients and find tools that allow the identification of “non-responders” early in the history of the disease.Research problem: PBC is characterized by immune dysregulation and increasing evidence suggests that innate immune cells play an active role early in the disease. Despite recent studies demonstrated a central role for Natural Killer (NK) cells in the destruction of BECs, a deep and comprehensive characterization of these cells in PBC patients is still lacking, especially in those patients that fails UDCA treatment. Methodology: In this project, we characterized the transcriptomic profile of circulating NK cell subsets in PBC patients and healthy donors (HDs) by single-cell RNA sequencing (scRNAseq); we also investigated differences in the transcriptomic profile of NK cells from PBC patients stratified based on their response to UDCA. By using a 23-color flow cytometry panel, we characterized and compared the frequencies and phenotype of NK cell subsets in the PB and liver from PBC patients and HDs. We also investigated differences in the immunophenotype of circulating NK cells from PBC patients based on their response to UDCA. We also set-up an “in vitro” experimental model to investigate the pathogenesis of the disease, by obtaining iPSC-derived BECs to be co-cultured with autologous PBMCs and LMNCs from end-stage “non-responder” patients and HDs.Key results: When we evaluated the transcriptomic profile of PB NK cells from PBC patients, we observed an overall activated transcriptomic profile compared with HDs, suggestive of higher cytotoxic activity, production of lytic granules and migration to inflammatory sites. We confirmed these results by flow cytometry, demonstrating a higher expression of activation markers and chemokine receptors on circulating NK cells from PBC patients than HDs. We also demonstrated that all subsets of NK cells in “non-responder” patients were characterized by a transcriptomic signature suggestive of higher cellular activation, inflammation, cytotoxic functions and pro-fibrotic profile compared with “responder” patients. Notably, we confirmed an overall activation of PB NK cells from “non-responder” patients also when we analyzed the NK cell phenotype by flow cytometry, as assessed by both manual gating strategies and unbiased PhenoGraph analysis. When we applied PhenoGraph analysis in the liver, we observed that the liver of end-stage “non-responder” PBC patients was infiltrated by a high proportion of liver resident (lr) CD56Bright NK cells that may be likely actively involved in the process of liver cirrhosis though the release of inflammatory cytokines. We also identified a cluster of adaptive-like lr-CD49a+ CD56Bright NK cells enriched in PBC patients that have been reported to have low cytotoxic properties but high cytokine production activity upon stimulation.Conclusions: By demonstrating an activated and pro-fibrotic profile of NK cells in PBC patients who failed first line treatment with UDCA, the results of this study suggest that NK cells, and in particular the CD56Bright NK cell subset, may play a relevant role in the active uncontrolled disease like"
ROLE OF NATURAL KILLER CELLS IN THE PATHOGENESIS OF PRIMARY BILIARY CHOLANGITIS / S. Franzese ; tutor: S. Della Bella ; director: N. Landsberger. Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, 2024 Jul 22. 36. ciclo
THE DIMENSIONS OF VARIABILITY MODELING OR, ON THE DESIGN OF SOFTWARE AND LANGUAGE PRODUCT LINES
Modern software systems must fulfill the needs of an ever-growing customer base. Due to the innate diversity of human needs, software should be customizable and reconfigurable. During the last decades, researchers and practitioners gained interest in Software Product Lines as an engineering technique suited to manage the complexity of highly-variable software systems without resorting to the clone-and-own approach, which is empirically proven to reduce the maintainability of software assets. This dissertation discusses several design aspects of Software Product Lines, with particular attention dedicated to the topic of Language Product Lines, i.e., Software Product Lines for the creation of families of programming language compilers and interpreters. The dissertation will overview the entire life-cycle of a Language Product Line: conception, creation, development, testing, deployment and maintenance. Each phase requires the proper tools and techniques, including an evaluation framework for the assessment of the quality of the results.
THE DIMENSIONS OF VARIABILITY MODELING OR, ON THE DESIGN OF SOFTWARE AND LANGUAGE PRODUCT LINES / L. Favalli ; tutor: W. Cazzola ; phd school headmaster: R. Sassi. Dipartimento di Informatica Giovanni Degli Antoni, 2023 Apr 27. 35. ciclo, Anno Accademico 2022.
THE POLITICISATION OF SOCIAL EUROPE. CONFLICT DYNAMICS IN THE POST-CRISIS DEBATE OVER EU SOCIAL AND EMPLOYMENT POLICIES
The bulk of the literature on ‘Social Europe’ has described the latest development of the social dimensions of the E(M)U in negative terms by referring to the absorption, the displacement, the decline and the marginalization of the EU social policies. Notably, the ruling of the European Court of Justice and the new post-crisis governance of the Economic and Monetary Union have contributed to increase the centrality of social issues in the European public debate, exacerbate the conflicts between political actors on EU social and employment policy and mount the dissensus towards the European integration process. While academic literature has broadly focused on the mobilization of political parties in defense of national welfare states against the EU “intrusiveness” into domestic decision-making and on the heightened politicization of EU affairs at the domestic level, less attention has been paid to the ‘politicization of Social Europe’, and especially to the configuration of the political conflicts over social integration at the EU level. Indeed, traditional literature on dimensions of politics in the European Parliament and the Council has ignored the specific conflict dynamics that characterize the political debate in this specific policy area. Therefore, the objective of this thesis is to fill this gap, and especially so by focusing on the new conflict constellations that emerged in the aftermath of the EU crisis. Drawing on the “clash syndrome” theory elaborated by Ferrera, I argue that the political debate over EU social integration is characterized by the overlapping of four lines of conflict of a functional, normative and territorial nature. Contrary to traditional literature, which has described the conflict over EU integration as mainly one- or bi-dimensional, the main finding of this thesis is that the political confrontation over EU social and employment policy is characterised by the coexistence of multiple and criss-crossing divides, which differently combine according to the arena where the debate takes place, the actors involved, the rules of the decision-making process and the issue at stake. The way these political divides interact leads to the creation of different conflict constellations, which can hinder the adoption of specific social policy proposal, but that can also open possibility spaces for the emergence of new coalitions that facilitate the adoption of an ambitious Social policy agenda.
THE POLITICISATION OF SOCIAL EUROPE. CONFLICT DYNAMICS IN THE POST-CRISIS DEBATE OVER EU SOCIAL AND EMPLOYMENT POLICIES / F. Corti ; tutor: M. Ferrera ; coordinatore NASP: M. Jessoula. Dipartimento di Scienze Sociali e Politiche, 2020 May 21. 32. ciclo, Anno Accademico 2019. [10.13130/corti-francesco_phd2020-05-21].
TRACTABLE DEPTH-BOUNDED APPROXIMATIONS TO SOME PROPOSITIONAL LOGICS. TOWARDS MORE REALISTIC MODELS OF LOGICAL AGENTS.
The depth-bounded approach seeks to provide realistic models of reasoners. Recognizing that most useful logics are idealizations in that they are either undecidable or likely to be intractable, the approach accounts for how they can be approximated in practice by resource-bounded agents. The approach has been applied to Classical Propositional Logic (CPL), yielding a hierarchy of tractable depth-bounded approximations to that logic, which in turn has been based on a KE/KI system. This Thesis shows that the approach can be naturally extended to useful non-classical logics such as First-Degree Entailment (FDE), the Logic of Paradox (LP), Strong Kleene Logic (K3) and Intuitionistic Propositional Logic (IPL). To do this, we introduce a KE/KI-style system for each of those logics such that: is formulated via signed formulae, consist of linear operational rules and branching structural rule(s), can be used as a direct-proof and a refutation method, and is interesting independently of the approach in that it has an exponential speed-up on its tableau system counterpart. The latter given that we introduce a new class of examples which we prove to be hard for all tableau systems sharing the ∨/∧ rules with the classical one, but easy for their analogous KE-style systems. Then we focus on showing that each of our KE/KI-style systems naturally yields a hierarchy of tractable depth-bounded approximations to the respective logic, in terms of the maximum number of allowed nested applications of the branching rule(s). The rule(s) express(es) a generalized rule of bivalence, is (are) essentially cut rule(s) and govern(s) the manipulation of virtual information, which is information that an agent does not hold but she temporarily assumes as if she held it. Intuitively, the more virtual information needs to be invoked via the branching rule(s), the harder the inference is for the agent. So, the nested application of the branching rule(s) provides a sensible measure of inferential depth. We also show that each hierarchy approximating FDE, LP, and K3, admits of a 5-valued non-deterministic semantics; whereas, paving the way for a semantical characterization of the hierarchy approximating IPL, we provide a 3-valued non-deterministic semantics for the full logic that fixes the meaning of the connectives without appealing to “structural” conditions. Moreover, we show a super-polynomial lower bound for the strongest possible version of clausal tableaux on the well-known class of “truly fat” expressions (which are easy for KE), settling a problem left open in the literature. Further, we investigate a hierarchy of tractable depth-bounded approximations to CPL based only on KE. Finally, we propose a refinement of the p-simulation relation which is adequate to establish positive results about the superiority of a system over another with respect to proof-search.
TRACTABLE DEPTH-BOUNDED APPROXIMATIONS TO SOME PROPOSITIONAL LOGICS. TOWARDS MORE REALISTIC MODELS OF LOGICAL AGENTS / A.j. Solares Rojas ; supervisore: A. Zamansky, M. D'Agostino ; coordinatore: A. Pinotti. Dipartimento di Filosofia Piero Martinetti, 2022 Jul 15. 34. ciclo, Anno Accademico 2021.
ANALYSIS OF THE STRIP OPTICAL SYSTEM FOR CMB POLARIZATION MEASUREMENTS
Nowadays, the detection of B-mode polarization anisotropies on large angular scales in the Cosmic Microwave Background polarization pattern is one of the major challenges in observational cosmology. In fact, it would give us an evidence in favor of the inflationary paradigm, shedding light on the physics of the very early Universe. Unfortunately, the amplitude of the signal is extremely low, at fractions of µK, and its detection requires high sensitivity instruments with thousands of detectors, a rigorous control of systematic effects, and a precise knowledge of the foreground polarized emission of our Galaxy.The Large Scale Polarization Explorer is one of the upcoming experiments devoted to the observation of the CMB polarization on large angular scales. It is based on two independent instruments: the STRIP ground-based telescope and the SWIPE balloon-borne mission.This thesis has been carried out in the context of the development of the STRIP instrument, a coherent polarimeter array that will observe the microwave sky from the Teide Observatory in Tenerife in two frequency bands centred at 43 and 95 GHz through a dual-reflector crossed-Dragone telescope of 1.5 m aperture. We present the implementation of the realistic electromagnetic model of the STRIP telescope, which includes all its optical elements and the surrounding structures. Since accurate predictions and measurements of the beam shape are essential both during the instrument design phase and for an in-depth knowledge of the whole-instrument response, we performed detailed optical simulations of the beam radiation properties. The results have been essential to assess the impact of the optical response on CMB polarization measurements when observing the sky with the nominal scanning strategy. This activity was mandatory because the optics is one of the major limiting factors for high precision measurements.
ANALYSIS OF THE STRIP OPTICAL SYSTEM FOR CMB POLARIZATION MEASUREMENTS / S. Realini ; director of the school: M. Paris ; supervisor of the thesis: A. Meniello ; cosupervisor of the thesis: C. Franceschet. Dipartimento di Fisica Aldo Pontremoli, 2020 Nov 18. 33. ciclo, Anno Accademico 2020. [10.13130/realini-sabrina_phd2020-11-18].
DOCTORAL DISSERTATION ON SUBCRESTAL IMPLANT PLACEMENT: ANALYSIS OF MARGINAL BONE LEVELS RELATED TO SURGICAL AND PROSTHETIC FACTORS
ABSTRACTBACKGROUND: The subcrestal placement of dental implants is becoming increasingly common. Modern biomedical devices with features to make them employable in such a manner have been developed to reduce the implant exposure within the oral environment. Indeed, it is universally recognized that, after placement, dental implants undergo a process of bone remodeling that has the greatest clinical impact at the level of the bony ridge. The exposure of the implant surface into the oral environment is potentially harmful because the accumulation of bacterial plaque on such surfaces can trigger inflammatory processes that, over time, may potentially lead to the loss of the implant itself. Subcrestal placement is based on the rationale that, provided a certain amount of crestal bone resorption cannot be avoided, this phenomenon remains superficially confined. According to this, the implant remains integrally within the bone envelope. Nevertheless, not all commercially available implant systems can be used in such a manner. Moreover, there are only a few clinical studies with short-to-medium-term follow-up in the literature. The primary outcome of this thesis is to investigate the prosthetic and surgical factors that may influence MBL of dental implants designed to be placed subcrestally. OBJECTIVES: General objectives of this thesis are: to present the clinical validation of subcrestal implants and to identify surgical and prosthetic factors that may affect MBL. Secondarily, the relationship between subcrestal placement and peri-implant biometric parameters is evaluated. Specific objectives of each study: (I.) To retrospectively evaluate 410 subcrestal dental implants placed in 93 patients in order to identify potential factors associated with MBL variation by multiple linear regression analysis. (II.) To investigate a possible correlation between the prosthetic emergence angle (EA) (greater or less than 30 degrees) of fixed rehabilitations screwed onto subcrestal implants and the peri-implant probing depth. Secondarily, to evaluate the correlation between emergence angle and gingival and plaque indices. (III.) To retrospectively evaluate the impact of vestibular emergence angle (EA) in the anterior zone with respect to peri-implant biometric indices of subcrestally placed implants. (IV.) systematic review of the literature to investigate the role of emergence angle (EA) on the development of peri-implantitis; (V.) A randomized controlled trial (RCT) with 3-year follow-up was conducted in order to evaluate the MBL of subcrestal implants placed by static computer-guided surgery versus freehand placement. Secondarily, to evaluate biological and prosthetic complications as well as peri-implant biometric parameters.MATERIALS & METHODS, RESULTS: Study IV.: A narrative review of the literature was performed to investigate the role of implant emergence angle on the development of peri-implantitis. Electronic search was performed on MEDLINE (PubMed) and Scopus by combination of MeSH terms and free text words combined through Boolean operators (AND or OR). Only English-language studies published up to June 2022 were selected. Prospective studies, retrospective studies, and case series with at least 10 patients were considered as eligible. All type of fixed implant-supported rehabilitations were evaluated: implant-supported single crowns, fixed partial restorations, and fixed complete restorations. By electronic search, 264 articles were identified; duplicate articles and those having only the abstract were removed. After title/abstract screening and full-texts reading, 5 studies were included. The studies selected for review employed different methods for measuring implant emergence angle. Katafuchi and Yi measured the angle of emergence (EA) radiographically from the mesio-distal aspect. The result was a higher prevalence of peri-implantitis
DOCTORAL DISSERTATION ON SUBCRESTAL IMPLANT PLACEMENT: ANALYSIS OF MARGINAL BONE LEVELS RELATED TO SURGICAL AND PROSTHETIC FACTORS / A. Palazzolo ; tutor: E. Romeo ; director: M. Del Fabbro. Dipartimento di Scienze Biomediche, Chirurgiche ed Odontoiatriche, 2024 Jul 01. 36. ciclo, Anno Accademico 2022/2023.
REDEFINING PROMOTER DNA METHYLATION CHANGE IN CANCER
Promoter DNA methylation has been recognized for decades as one of the majormechanisms of epigenetic gene regulation and has consistently been found to be perturbedduring cancer development. However, among the many genomic regions that have beenfound to display altered methylation in different cancer types, exactly which changes play acausative role in cancer development and how they impact the cancer transcriptomeremains poorly understood. We developed two computational tools, MethylDriver andMethodical, to advance our understanding of the relationship between DNA methylationand transcription and how this relationship is perturbed in cancer and to identifymethylation changes that are most likely playing a causative role in cancer development.MethylDriver leverages epigenetic covariates to separate promoter methylation changeswhich may be under selection and playing a causative role during tumour progression fromthe many changes which are likely incidental to cancer development. We appliedMethylDriver to 13 different cancer types from The Cancer Genome atlas (TCGA) toidentify putative epigenetic drivers in a variety of cancer types, uncovering relatively smallsets of differentially methylated promoters with increased relevance to cancer biology.Despite these findings, we were surprised by the generally poor correlations we observedbetween promoter DNA methylation and transcriptional expression of their associatedgenes and also by the lack of consensus for the general extent of promoter regions aroundtranscriptional start sites (TSS). We thus developed Methodical, a computational methodthat combines RNA-seq and WGBS data to identify genomic regions where DNAmethylation tends to be highly correlated with nearby TSS activity. We refer to theseregions as TSS-proximal methylation-controlled regulatory sites (TMRs). We appliedMethodical to one normal prostate tissue data set, one prostate tumour dataset and oneprostate metastases dataset and characterized the identified TMRs. We reveal that, incontrast with the conventional idea that DNA methylation silences transcription, asubstantial minority of TMRs display positive methylation-transcription correlations. Wealso show that the region just downstream of the TSS is the most common location forTMRs and that negative and positive TMRs are enriched for different genomic features and8chromatin states. Finally, we demonstrate that TMRs are among the regions most affectedby DNA methylation change in diverse cancer types.
REDEFINING PROMOTER DNA METHYLATION CHANGE IN CANCER / R.o. Heery ; tutor: M. Schaefer, S. Santaguida ; phd coordinator: S. Minucci. Dipartimento di Oncologia ed Emato-Oncologia, 2024 Apr 11. 35. ciclo, Anno Accademico 2023.
A FULL-STACK SOFTWARE ARCHITECTURE FOR THE CONTROL AND CALIBRATION OF QUANTUM HARDWARE
Over the last 20 years, thanks to the development of quantum technologies, it has been possible to deploy quantum algorithms and applicationsthat before were only accessible through simulation on real quantumhardware. The current devices available are often referred to as noisyintermediate-scale quantum (NISQ) computers, and they require calibration routines in order to obtain consistent results.In this context, we present Qibo, an open-source framework for quantum computing. Qibo was initially born as a tool for simulating quantumcircuits. Through its modular layout for backend abstraction, it is possibleto change effortlessly between different backends, including a simulatorbased on just-in-time compilation, Qibojit.In order to enable the execution and calibration of self-hosted quantumhardware we have developed two open-source libraries integrated withthe Qibo framework: Qibolab and Qibocal. Qibolab provides the software layer required to automatically execute circuit-based algorithmson custom self-hosted quantum hardware platforms. It enables experimentalists and developers to delegate all complex aspects of hardwareimplementation to the library so they can standardize the deployment ofquantum computing algorithms in a hardware-agnostic way.Qibocal is based on a modular QPU (Quantum Processor Unit) platform agnostic approach and introduces tools that support the calibrationand characterization of QPUs on three different levels: development,deployment and distribution. Qibocal provides a code library to rapidlydevelop protocols for different hardware abstraction layers. The integration with Qibo allows one to easily switch between hardware executionand high-perfomance simulation.Finally, to showcase the capabilities of the framework, we present thecalibration of a superconducting qubit pair with Qibocal and applicationsdeveloped with Qibo, including a novel method for improving groundstate preparation through Variational Quantum Eigensolvers and DoubleBracket algorithms.
A FULL-STACK SOFTWARE ARCHITECTURE FOR THE CONTROL AND CALIBRATION OF QUANTUM HARDWARE / E. Pedicillo ; supervisor: S. Carrazza ; coordinator: A. Mennella. - Università degli Studi di Milano. Dipartimento di Fisica Aldo Pontremoli, 2025 Nov 28. 38. ciclo, Anno Accademico 2024/2025.
PNPLA3 P.I148M VARIANT INCREASES LIPID DROPLETS NUMBER AND SIZE IN PATIENT-DERIVED LIVER ORGANOIDS
PNPLA3 rs738409 C>G p.I148M variant is the main genetic determinant of metabolic associated steatotic liver disease (MASLD). To this day, this association still lacks a molecular characterization, due to the lack of comprehensive human models. To overcome this gap in knowledge, we generated patient-derived human liver organoids (HLOs) and studied lipid accumulation across different PNPLA3 genotypes. HLOs were differentiated toward a hepatocyte-like phenotype (Hep-HLO) and exposed to 300 µM Palmitic and Oleic fatty acids (FAs) for 72h. Lipid droplets (LDs) average size and number (normalized for nuclei) were quantified by NileRed staining and compared between each PNPLA3 genotype.Regardless of FA treatment, M/M Hep-HLOs show increased LDs size when compared to I/M and I/I. Conversely, LDs number and total area showed direct correlation with the number of mutated alleles, indicating that the p.I148M substitution promotes the formation of new LDs, which was exacerbated by exposure to FAs. Overall, these data confirm on novel 3D cellular models the seminal role of rs738409 in the development of MASLD and suggest that LDs size and number are regulated by independent mechanisms.Preliminary lipidomic analyses highlighted an enrichment in sphyngomyelins, lysophosphatidylcholines and ceramides in PNPLA3 M/M Hep-HLO, which was exacerbated by FAs administration. These lipid species are associated with cellular damage, inflammation, Insulin resistance, oxidative stress and MASH development. The opposite trend was observed for phosphatidylcholines, which are essential for LDL loading and triacylglycerols export from the liver and whose levels drop during MASLD development; highlighting an unbalance towards lipid species that are proinflammatory and pro-apoptotic in Hep-HLO homozygous for PNPLA3 rs738409. Overall, these data corroborate the robustness of Hep-HLO as a preclinical in vitro model for genetic predisposition for MASLD. We are now developing siRNAs specific for silencing PNPLA3 rs738409 transcripts to examine the impact on LD phenotypes.
PNPLA3 P.I148M VARIANT INCREASES LIPID DROPLETS NUMBER AND SIZE IN PATIENT-DERIVED LIVER ORGANOIDS / E. Casirati ; TUTOR: Prof. LUCA VITTORIO CARLO VALENTICO-TUTOR: Dr. ALESSANDRO CHERUBINICOORDINATORE DEL DOTTORATO: Prof.ssa CHIARELLA SFORZA. Università degli Studi di Milano, 2024 Jun 17. 36. ciclo, Anno Accademico 2023.
INTEGRATIVE GENOMICS UNVEILS TRANSCRIPTION FACTOR ROLES IN CELLULAR FATEAND SPATIALLY RESOLVED TRIPLE-NEGATIVE BREAST CANCER IMMUNITY CAPACITY
In this thesis, we explore integrative genomics through advanced sequencing to understand biological systems and processes, focusing on two main projects. The first investigates transcription factors (TFs) in cellular differentiation, addressing the inefficiency in current cellular conversion strategies due to the complex TF regulatory landscape. We developed a comprehensive screening of 130 developmental TFs, comparing their dosages in a single experiment to identify novel pioneer factors crucial for cellular identity. This approach aims to enhance our understanding of cell fate decisions. The second project examines triple-negative breast cancer (TNBC), characterized by its aggressiveness and treatment challenges due to tumor heterogeneity. We propose a novel clinical workflow combining spatial transcriptomics, RNA sequencing, and Immunohistochemistry to analyze TNBC, focusing on PD-L1 status. This method revealed sub-tumoral variations and identified LY6D as a new diagnostic marker, highlighting the potential of spatial transcriptomics in personalized medicine for TNBC. Overall, this dissertation contributes to the fields of cellular reprogramming and cancer diagnostics by integrating various genomic data and technologies to uncover complex molecular mechanisms and identify new therapeutic targets.
INTEGRATIVE GENOMICS UNVEILS TRANSCRIPTION FACTOR ROLES IN CELLULAR FATEAND SPATIALLY RESOLVED TRIPLE-NEGATIVE BREAST CANCER IMMUNITY CAPACITY / S. Slovin ; supervisor: D. Cacchiarelli ; internal advisor: G. Gambardella ; external advisor: P. Carninci. - English. Dipartimento di Oncologia ed Emato-Oncologia, 2024 Apr 22. 35. ciclo, Anno Accademico 2023.
EVOLUTION EQUATIONS WITH APPLICATIONS TO POPULATION DYNAMICS
The main topic of this thesis is the analysis of evolution equations reflecting issues in ecology and population dynamics. In mathematical modelling, the impact of environmental elements and the interaction between species is read into the role of heterogeneity in equations and interactions in coupled systems. In this direction, we investigate three separate problems, each corresponding to a chapter of this thesis. The first problem addresses the evolution of a single population living in a periodic medium with a fast diffusion line; this corresponds to the study of a reaction-diffusion system with equations in different dimensions. We derive results on asymptotic behaviour through the study of some generalised principal eigenvalues. We find that the road has no impact on the survival chances of the population, despite the deleterious effect expected from fragmentation. The second investigation regards a model describing the competition between two populations in a situation of asymmetrically aggressive interactions; this consists of a system of two ODEs. The evolution progresses through two possible scenarios, where only one population survives. Then, the interpretation of one of the parameters as the aggressiveness of the attacker population naturally raises questions of controllability. We characterise the set of initial conditions leading to the victory of the attacker through a suitable (possibly time-dependant) strategy. The third and last part of this thesis analyses the time decay of some evolution equations with classical and fractional time derivatives. Depending on the type of derivative and some degree of non-degeneracy of the spatial operator, quantitative polynomial or exponential estimates are entailed.
EVOLUTION EQUATIONS WITH APPLICATIONS TO POPULATION DYNAMICS / E. Affili ; tutor: E. Valdinoci, L. Rossi ( Sorbonne Universite'). Università degli Studi di Milano, 2021 Mar 11. 33. ciclo, Anno Accademico 2020. [10.13130/affili-elisa_phd2021-03-11].
SURVIVING UNDER PRESSURE: TRANSLATIONAL REWIRING IN RESPONSE TO MECHANICAL STIMULI
An outcome of high mechanical stress in the context of hyperproliferation and increased cellular tension is the reduction in cell volume, that holds the potential to impact cellular density. Both cell size and density homeostasis are integral features of healthy tissues, intricately linked to the regulation of the cell cycle and biosynthesis processes. This connection suggested that achieving density homeostasis could involve a coupling mechanism between the regulation of macromolecular production and osmolytes. Considering the importance of density and protein homeostasis, we decided to explore whether similar mechanisms may play a role in the responses of tumor cells upon mechanical stress.To shed light on the mechanisms, we have employed several approaches to study the acute alterations in cell size induced by mechanical stress. We used osmotic shocks and compressive stresses, as well as genetic encoded microparticle (GEMs) tracking to infer cytoplasmic density. The combination of these tools with measures of translation rate, polysome profiling, ribogenesis, and protein degradation assays uncovered interesting evidences demonstrating a mechanism that couples the regulation of macromolecular production with rheological adaptation in colorectal cancer cells HCT116. In particular, we have found that the early response to mechanical compression is independent of the mechanistic target of rapamycin complex 1 (mTORC1), while adaptation requires mTORC1 inactivation and subsequent increase of autophagy. Our data provided clues suggesting alternative signaling and biophysical pathways involved in the regulation of protein homeostasis in response to mechanical stress, which provide valuable insights into the pathogenesis of cancer and may represent promising targets for therapeutic intervention.
SURVIVING UNDER PRESSURE: TRANSLATIONAL REWIRING IN RESPONSE TO MECHANICAL STIMULI / G. Fragale ; tutor: K. Havas ; co-tutor: M. Foiani. Dipartimento di Oncologia ed Emato-Oncologia, 2024 Apr 11. 35. ciclo, Anno Accademico 2023.
PARADIGMS OF EVIDENCE. REFRAMING FORENSIC IMAGES THROUGH EXTENDED REALITY MEDIA
This research explores contemporary transformations of visual evidence in the context of anexpanding visual culture shaped by hybridisations between photography and extended realitymedia. Starting from the historical centrality of photography as a medium of evidence, traditionallybased on notions of transparency and mechanical objectivity, the thesis develops a reflection basedon Carlo Ginzburg's evidential paradigm, which is reinterpreted in terms of a forensic gaze. Thisparadigm allows evidence to be conceived not simply in terms of the apparent absence ofmediation, but as a clue to be interpreted and an operational tool for investigation.Starting from this theoretical framework, the research examines the forensic uses of photographyfrom the nineteenth century to the present day, identifying a genealogy of XR media in thecartographic and metric tradition of crime scene documentation. Through the analysis ofcontemporary investigative practices, in particular those developed by Forensic Architecture, itshows how virtual reconstructions do not mark a break with photographic images, inviting anintermedial reconceptualisation of visual evidence that goes beyond the specificity of thephotographic medium.This theoretical framework is then tested through archival research conducted at the Archivesnationales in Paris, focusing on forensic images used in the so-called V13 trial, i.e. the legal trialfollowing the 13 November 2015 attacks in Paris. The thesis analyses the institutional, archival andmedia conditions under which forensic images were presented in court, situating their presence inthe courtroom through the semiopragmatic device of recontextualisation. The research highlightsthe tensions that arise when visual evidence of the attack scenes is shown in court: tensions betweenvisibility and concealment, between testimonial function and control logic, between renunciationand remediation.Finally, the thesis concludes with an art-based research experiment that explores the audiovisualarchive of the V13 trial in light of its difficult accessibility through an immersive installation.Developed from interviews with archivists and spatialised through a generative sound score andphotogrammetric scans of archival working documents, the work evokes a documentary heritage bysummoning it, without showing it, in terms of “missing pictures”.
PARADIGMS OF EVIDENCE. REFRAMING FORENSIC IMAGES THROUGH EXTENDED REALITY MEDIA / R. Cinelli ; supervised by: B. Grespi, M. Treleani, M. Sobieszsanski ; coordinator: N. Guicciardini Salviati. Dipartimento di Filosofia Piero Martinetti, 2025 Jun 30. 37. ciclo, Anno Accademico 2024/2025.
SECURE PLANT FERTILITY IN A CHANGING CLIMATE
The megaspore mother cell (MMC) development is a crucial step for plant reproduction. MMC specification is regulated by hormones, and many genes are involved. SPOROCYTELESS/NOZZLE (SPL/NZZ) is considered the master regulator of this process since its mutant cannot specify the MMC. It encodes a transcriptional repressor protein, which is present in all land plants. Despite the importance of this gene, its role is not completely unveiled. Thanks to the work presented in this thesis, it is possible to propose a mechanism of action for SPL, which involves its interaction with MADS-domain transcription factors. In a complex with SEPALLATA3 (SEP3), SPL negatively regulates the expression of AINTEGUMENTA (ANT) in the nucellus. ANT is a repressor of the auxin transporter PIN1, in this way the ANT repression allows PIN1 expression in the nucellus. In this work, PIN1 is described to be fundamental for auxin transport at the tip of ovule primordia. Here, thanks to the ARF transcription factors, auxin promotes the differentiation of the MMC. The function of SPL seems to be conserved in plants. Indeed, preliminary data show that the protein encoded by the AtSPL homologous in Marchantia polymorpha, MpSPL, can rescue the MMC formation in Arabidopsis spl.1 mutant. The presented results show that the formation of the MMC is regulated by mechanism conserved in evolution and that, in Arabidopsis thaliana, it involves auxin transport and signalling, regulated by SPL and MADS-domain transcription factors.
SECURE PLANT FERTILITY IN A CHANGING CLIMATE / C. Astori ; tutor: L. Colombo ; supervisor: A. Cavalleri ; PhD course coordinator: S. Ricagno. Dipartimento di Bioscienze, 2025 Feb 27. 37. ciclo, Anno Accademico 2023/2024.
REPLICAS IN COMPLEX SYSTEMS: APPLICATIONS TO LARGE DEVIATIONS AND NEURAL NETWORKS
In this thesis, we apply methods from replica theory to deal with two contemporary problems in the study of complex systems. In Part I, we discuss the behavior of the rare fluctuations of the observable of choice in most models of spin glasses: the free energy. Due to the quenched disorder, this and other thermodynamic quantities are self-averaging random variables, whose probability distribution can be evaluated within replica theory. In particular, the probability of the free energy fluctuations above its typical value shows an anomalous scaling with the number of degrees of freedom, at variance with the ordinary exponential suppression for fluctuations below. We explain how the introduction of a small magnetic field can remove this anomalous behavior. In Part II, we apply the replica formalism to the problem of linear classification of objects with a geometrical structure, in the context of machine learning. In particular, using combinatorial techniques we evaluate the number of dichotomies (binary classifications) of a set of structured inputs achievable by a linear classifier, as a function of the number of inputs to classify. We prove that this number shows an additional critical point beyond the usual storage capacity for isolated points, at which the number of admissible dichotomies becomes zero in the thermodynamic limit; the associated phase transition present a certain degree of replica symmetry breaking. This behavior is due to a trade-off between the increasing number of points to classify, and the increasing volume excluded by their geometrical structure. This approach goes in the direction of finding bounds on the generalization error of certain simple neural network architectures, more stringent for structured data than the ones known from Statistical Learning Theory.
REPLICAS IN COMPLEX SYSTEMS: APPLICATIONS TO LARGE DEVIATIONS AND NEURAL NETWORKS / M. Pastore ; tutor: S. Caracciolo ; coordinatore: M. Paris. Università degli Studi di Milano, 2020 Oct 27. 33. ciclo, Anno Accademico 2020. [10.13130/pastore-mauro_phd2020-10-27].
CONFIGURATION MIXING INVESTIGATION IN GERMANIUM ISOTOPES
Protons and neutrons of atomic nuclei can arrange in both spherical and non-spherical configurations. Nuclei can also exhibit configurations which lie close in energy, yet present different deformations. Such phenomenon is referred to as shape coexistence and its occurrence in nuclei is thought to pervade the entire nuclear chart. Excited states with similar energies and same spin and parity can also mix, so that the observed states are the result of mixing of different configurations.In this context, the structure of Ge isotopes has been widely investigated over the years. Previous transfer reaction experiments pointed to the occurrence of configuration mixing between the ground state and first excited 0+ state in 72Ge (N = 40). Furthermore, they indicated a weakening of such mixing moving away from N = 40, in correspondence with the increase of the excitation energy of the 0+[2] state. However, the nature of the configurations that mix and their evolution along the Ge isotopic chain is still elusive. In this thesis, a study of configuration mixing in the even-even Ge isotopes at and near stability (A = 72, 74, 76, 78) has been carried out through the measurement of electric monopole (E0) transition strengths, which have been identified as a sensitive probe of configuration mixing. In particular, the focus has been set on the transitions with I = 2, 2+[2]→2+[1]. The experiments presented in this thesis have been performed at the TRIUMF laboratory (Vancouver, Canada). The isotopes of interest have been populated via the β decay of radioactive ion beams of 72,74,76,78Ga, which were implanted at the center of the GRIFFIN decay station. The GRIFFIN spectrometer, combined with the PACES silicon array, enabled us to perform both γ-ray and electron spectroscopic investigations, to measure E0 strengths between states with I = 2.
CONFIGURATION MIXING INVESTIGATION IN GERMANIUM ISOTOPES / C. Porzio ; supervisor: S. Leoni ; co-supervisor: A. Garnsworthy ; coordinatore: M. Paris. Università degli Studi di Milano, 2021 Dec 13. 34. ciclo, Anno Accademico 2021. [10.13130/porzio-carlotta_phd2021-12-13].
Emperor Honorius has not received much attention from historiography, which has often limited itself to emphasising his inadequacy and weakness in relation to the various problems that the western part of the empire was experiencing; indeed, barbarian incursions, complex religious issues, and the constant need for considerable economic resources were formally entrusted to the purple of a teenager, given that Honorius came to power in 395 when he was only eleven years old. In fact, it was others who influenced the political direction of Honorius' reign, among them Stilicho and Constantius III: the former as regent appointed by Theodosius I on his deathbed, the latter also appointed emperor in 421. This does not detract from the fact that the figure and reign of Honorius nevertheless deserves to be framed in the light of both the few literary testimonies we have (e.g. Zosimus, Claudian and the chronological sources) and, above all, through the more than four hundred and fifty constitutions issued by his chancellery in the twenty-eight years of his reign. In particular, the biography of the young emperor will be traced, highlighting the most salient and most debated moments in the literature, with particular reference to his childhood and the role of Stilicho. It has been decided to provide a list of all constitutions attributed to Honorius that are known from the Theodosian Codex and the Codex Iustinianus, giving their respective correspondences, a summary of their content and data on their addressee and dating.Two Appendices, dedicated respectively to the addressees of the constitutions and the places of promulgation, data that can be deduced from the inscriptiones and subscriptiones, conclude this section, to offer hints for prosopographical research and to assess the movements of the imperial comitatus.Then, a number of thematic in-depth studies were carried out: the first encompasses some interventions by Honorius' chancellery on African issues, both from an anti-heretical and anti-pagan perspective, as well as disciplining public order (discipline against raids by nomadic peoples and the case of Heraclianus). Finally, we wanted to identify some of Honorius' constitutions on the subject of succession in order to verify how the institutions and classical law were transposed and possibly adapted to new situations.Many of the major questions concerning late antique legislation stand out when dealing with Honorius' legacy of legislation: from the relationship between the West and the East to the choices of legislative technique oscillating between a concrete case and a tension towards generalisation, from the effects of the textual manipulations carried out by Theodosius II's commissioners to the manuscript tradition of the two Codices.
Ricerche sulla legislazione di Onorio / L. Lanti ; tutor: I. Fargnoli ; coordinatrice: F. Poggi. Dipartimento di Diritto Privato e Storia del Diritto, 2024. 36. ciclo
MOLECULAR CHARACTERIZATION OF STAPHYLOCOCCUS PSEUDINTERMEDIUS: RESEARCH ON VIRULENCE FACTORS, ANTIBIOTIC-RESISTANCE AND QUORUM-SENSING
Staphylococcus pseudintermedius (S. pseudintermedius) is one of the most important pathogens in small animal veterinary medicine. In the dog, it is a commensal bacterium able to cause severe diseases (mainly dermatological), which rapidly evolve to persistent and chronic infections. This pathogen is also considered a public health problem due to the rapid evolution and spread of antibiotic-resistant clones, in particular, the worldwide dissemination of Multi-Drug Resistant (MDR) strains and methicillin-resistant S. pseudintermedius (MRSP). Nowadays, its zoonotic potential is under constant investigations. The first chapter of the thesis centers the attention on the typing techniques used to study a population of S. pseudintermedius isolated from the skin of dogs and raw milk of Holstein-Fresian dairy cattle. The aims of this study were the investigation of the genetic relatedness of different S. pseudintermedius strains using three molecular typing techniques; the Ribosomal Spacers Amplification PCR (RSA-PCR), the Random Amplification of Polymorphic DNA PCR (RAPD-PCR) and the Restriction Fragment Length Polymorphism PCR (RAPD-PCR). Moreover, the dissemination of species-specific genes coding for exfoliative toxins (siet, expA, and expB), enterotoxin (seCcanine) and a bicomponent leukocidin (lukS-F) was studied to understand the virulence of the strains. The study was carried out on a final population of 40 S. pseudintermedius strains isolated from 42 clinical cases of deep canine pyoderma and from 5 samples of raw milk from Holstein-Fresian dairy cattle with intramammary infection. After the microbiological examination and phenotypic typing, 47 S. pseudintermedius strains were collected and genetically analyzed. RSA-PCR revealed the presence of a unique biotype. The dendrogram obtained from RAPD-PCR disclosed two important results: i) the staphylococcal strains isolated from dogs and cows are genetically similar underlying the ability of this pathogen to colonize also dairy cattle; ii) the presence of a single cluster grouping all the S. pseudintermedius strains. Based on the presence of a specific restriction site in the phosphotransacetylase (pta) gene, RFLP-PCR confirmed the presence of 40 S. pseudintermedius and 7 non-S. pseudintermedius strains. The sequencing unveiled the effective presence of one strain of S. aureus and one of E. faecalis, 5 strains of S. schleiferi and 40 of S. pseudintermedius. All the S. pseudintermedius isolates were positive for siet gene, while expA and expB were amplified by, respectively, 10% and 17,5%. Specific enterotoxin seCcanine and leukotoxin luk-S-F were found with a prevalence of 35% and 92,5%, respectively. Therefore, the combined use of molecular typing techniques and virulence factors screening allows to better characterize S. pseudintermedius and to understand its zoonotic potential. The second chapter points out the attention to the problem of antibiotic resistance. Frequently, in Veterinary and Human medicine too, wide-spectrum antibiotics are prescribed to face with bacteria-based diseases without the microbiological examination and relative antibiotic-resistance screening. This could lead to the selection and spread of antibiotic-resistant strains. The aim of this work was to study the antibiotic-resistance profile of a large population of S. pseudintermedius strains collected in two Italian veterinary teaching hospitals, the first in Milan (Università degli Studi di Milano) and the second in Naples (Università degli Studi di Napoli “Federico II”) One-hundred sixteen and one-hundred twenty-six strains were collected in Milan and Naples, respectively. A panel of 10 different antibiotic molecules was tested with the Kirby-Bauer assay, qualitative PCR was used to describe the dissemination of mecA and tetM/K genes. The results showed a high rate of resistance for amoxicillin+clavulanate, clindamycin, tobramycin and tetracycline also
MOLECULAR CHARACTERIZATION OF STAPHYLOCOCCUS PSEUDINTERMEDIUS: RESEARCH ON VIRULENCE FACTORS, ANTIBIOTIC-RESISTANCE AND QUORUM-SENSING / G. Meroni ; tutor: P. A. Martino ; coordinatore: V. Grieco. DIPARTIMENTO DI MEDICINA VETERINARIA, 2020 Feb 05. 32. ciclo, Anno Accademico 2019. [10.13130/meroni-gabriele_phd2020-02-05].
GENOMICS OF EFFICIENCY, RESILIENCE AND BIODIVERSITY IN HOLSTEIN AND LOCAL CATTLE BREEDS
This thesis explores the genomics of biodiversity, resilience, and efficiency in Holstein (a cosmopolitan population) and local cattle breeds, focusing on their genetic diversity, adaptation to environmental challenges and the application of genomic tools in breeding programs. The research primarily investigates genetic diversity among autochthonous cattle breeds in the Aosta Valley, particularly the Aosta Red Pied, Aosta Black Pied-Chestnut, and their subgroup Aosta Chestnut-Hérens, alongside the highly selected Holstein Friesian breed.The study leveraged genomic information such as Single Nucleotide Polymorphism (SNP) markers to assess genetic variation across breeds and conduct genome-wide association studies. Emphasis was placed on exploring the impact of genomic variations on traits linked to disease resistance (e.g., mastitis and bovine respiratory disease) and milk production efficiency. The Holstein breed, a specialised cosmopolitan population known for its high milk production and intensive farming system, served as a reference. In contrast, local breeds like Aosta were studied for their resilience and production efficiency in harsh environmental conditions, such as alpine mountain pastures.Key findings include the identification of Runs of Homozygosity (ROH), used to estimate inbreeding and genomic diversity and to map genomic regions under selection. The ROH analysis showed that local breeds possess unique adaptations, exhibiting lower inbreeding coefficients than the Holstein, which has faced a loss of genetic variation due to the intense selection that occurred for milk production traits in the early past. This thesis also mapped genomic regions associated with mastitis resistance in Holstein, contributing to better breeding strategies aimed at improving animal health and welfare.Additionally, this work highlights the critical role of local breeds in maintaining biodiversity. These breeds offer a source of genetic variation essential for sustainable cattle farming and their resilience could help in facing future environmental changes. The genetic makeup of local breeds also supports cultural and economic activities in the Aosta Valley, such as the production of Fontina cheese, a Product of Designated Origin (PDO), and the Bataille de Reines.Finally, this thesis proposes that coupling genomic data with phenotypic variability enhances the understanding of resilience traits in cattle. It suggests that local breeds, despite their small population size, are valuable resources in the face of global challenges such as climate change and the increasing demand for sustainable livestock systems.
GENOMICS OF EFFICIENCY, RESILIENCE AND BIODIVERSITY IN HOLSTEIN AND LOCAL CATTLE BREEDS / F. Bernini ; tutor: A. Bagnato ; coordinatore: F. Ceciliani. Dipartimento di Medicina Veterinaria e Scienze Animali, 2025 Mar 20. 37. ciclo, Anno Accademico 2023/2024.
HIGH-MOUNTAIN ENVIRONMENT UNDER CLIMATE CHANGE, THE INTERACTIONS BETWEEN CLIMATE, METEOROLOGY AND GEOMORPHOLOGICAL FACTORS
The Alps, and in general the high mountain areas, in recent decades are under going profound changes due to climate change. The peculiarities and the greater vulnerability of these territories, characterized by complex and delicate balances, are threatened by global warming. In fact, it considerably reduces the habitat of many plant and animal species, pushing them towards ever greater elevations. Once this living zones have reached the tops of the reliefs may no longer be available, and may lead to the disappearance, from these environments, of endemic biodiversity (Hannah et al., 2002). At the same time, however, the progressive debris coverage of the glacial tongues (Deline, 2005; Caccianiga et al., 2011) and the progressive expansion of the proglacial areas, offer new spaces and areas that can be colonised by the vegetation systems and animal communities (Whittaker, 1993).The changes underway in these areas not only affect biological systems, but also geodiversity and the geoheritage of the alpine and mountain environment. “Geodiversity” can be defined simply as “the natural range (diversity) of geological (rocks, minerals, fossils), geomorphological (land form, physical processes) and soil features. It includes their assemblages, relationships, properties, interpretations and systems” (Gray, 2004). For definition of “geodiversity” it follows that an impact on geomorphological processes can affect the local diversity of an area. Examples are the changes in the speed and intensity of processes, such as for example the acceleration of glacial shrinkage, the degradation of permafrost (Deline et al., 2015), the impact of the action of running waters connected to extreme events, the mass wasting processes (Cossart and Fort, 2008; Lane et al., 2017) and the related interactions with biological systems (Eichel, 2015).More in detail in the Alps, but as happens in other mountain ranges, glaciers are shrinking and retreating at ever increasing levels, losing the valley tongues and more and more extensive areas are leaved by the ice. The newly formed proglacial zones are characterized by an abundance of incoherent debris, easily dislocated by the running waters, especially during intense meteorological phenomena (Luino, 2005). These areas, in many cases easily accessible by mountain-goers and not only by expert mountaineers, may therefore present geomorphological dangerous situations, increased by the extremes of meteorological events (for example drought or intense precipitation events). This entails an increase in risks for tourist and even productive activities. The increase in the risk factor for hikers can also derive not only from an increase in danger, for example along the paths, but also from an increase in vulnerability given by sudden changes in weather conditions and the greater difficulty of walking on the paths (Fort, 2015; Pelfini et al., 2007a).The current studies must face the challenge of knowing more in detail the complex interactions that take place in these changing territories, and moreover it is needed to at least partly predict their evolution and behaviour. This challenge has a double value, both scientific, and therefore to broaden the knowledge on the complexity of nature, and civil, because the knowledge of phenomena and the environment allows a more conscious attendance of the high mountain territories. This is the first step for the mitigation of the risk, furthermore, a development that is as environmentally sustainable as possible.Certainly in the coming decades the Alps landscapes will be profoundly different from those of the last century. The progressive reduction of the cryosphere will also have repercussions on many aspects of everyday life of the population that lives not in close contact with the ice giants, but also at a considerable distance from the glacial areas. For example, it has to be considered the availability of drinking water,
HIGH-MOUNTAIN ENVIRONMENT UNDER CLIMATE CHANGE, THE INTERACTIONS BETWEEN CLIMATE, METEOROLOGY AND GEOMORPHOLOGICAL FACTORS / A. Golzio ; supervisors: M. Pelfini, M. Maugeri ; coordinator: G. F. Ficetola ; curators: G. Diolaiuti, C.Cassardo, S. Ferrarese, M. Manfrin. Dipartimento di Scienze della Terra Ardito Desio, 2020 Feb 10. 32. ciclo, Anno Accademico 2019. [10.13130/golzio-alessio_phd2020-02-10].
SOFT POLYMERIC DEVICES FOR MORPHO-FUNCTIONAL MIMICRY OF BIOLOGICAL STRUCTURES
In the time-lapse of hundreds of millions of years, living organisms have continuously evolved, developing a myriad of astonishing forms and abilities to be often inspirational for mankind. In 1997, Janine Benyus, paved the onset of a revolutionary discipline called biomimicry. Biomimicry can be elegantly defined as “innovation inspired by nature”. From a practical point of view, it is an approach to innovation that aims to solve human challenges by emulating nature’s time-tested strategies. A range of technological improvements have been inspired by terrestrial life and examples of biomimicry now include synthetic materials with new mechanical properties that emulate the brick-and-mortar architecture found in bone and mollusc shells; nanoparticles and cell-based therapies with strong potential for the advancement novel and more effective therapeutic and imaging systems; high-speed trains with a nose emulation the kingfisher bird’s beak, as a way of eliminating the buildup of atmospheric pressure when passing through tunnels; and renewable energy production by using bio-engineered bacteria capable of synthesising almost one thousand litres of hydrogen gas per hectare.We are at the threshold of a new era for science and engineering in which biologically inspired designs offer novel and sustainable solutions to society’s problems that may not be provided as quickly or economically by traditional paths. To address this demand, there is a continual quest to seek new materials and fabrication techniques with unprecedented integration of properties and functionalities at minimal production costs. My PhD research work focused on the development of materials, manufacturing processes and devices that aim at solving current bottlenecks in the fields of soft robotics, tissue engineering and environmental control of harmful insects.
SOFT POLYMERIC DEVICES FOR MORPHO-FUNCTIONAL MIMICRY OF BIOLOGICAL STRUCTURES / M. Piazzoni ; supervisor: C. Lenardi ; director of the school: M. Paris. Dipartimento di Bioscienze, 2021 Dec 13. 34. ciclo, Anno Accademico 2021.
NEW RESEARCH ON AGROTECHNICS FOR ORGANIC RICE PRODUCTION AND THEIR SUSTAINABILITY ASSESSMENT
"This thesis studies and evaluates the agronomical techniques associated with organic rice cultivation in the Northern Italy area. The study funding is from the Italian MIPAAF project ""Risobiosystems"", which started in 2017 and ended in 2020.The studies presented are connected to the European sustainable food production strategy ""Farm to Fork,"" which is the core of the European Green Deal to make food systems fair, healthy and environmentally friendly, encouraging the expansion of the organic agriculture sector (European Commission 2020). As reported in the literature, organic farming is capable of reducing the environmental impact of agriculture by avoiding the use of synthetic compounds (e.g. fertilisers, pesticides) and by promoting practices (e.g. crop rotation, leguminous cultivation, organic fertilisers, green manure crops, green mulching.) able to increase the soil carbon stock, and prevent the indirect environmental impacts due to the industrial production of inputs (Acuna et al. 2018). Focusing on the rice sector, the organic system was observed able to increase the soil carbon storage capacity (Komatsuzaki and Syuaib, 2010) and organic matter content, facilitating the soil preparation (Mendoza, 2004) and promoting the ecological succession and temporal heterogeneity of the macrophyte communities into the soil (Martínez- Eixarch et al.; 2017). The thesis is a collection of articles published or under review associated with the organic rice production connected with Risobiosystems project results. Each paper explains my work and contribution, and each Chapter gave rise to new and ongoing studies.Chapter I (https://doi.org/10.1016/j.agsy.2019.102739) reveals the lack of scientific research and information about organic rice farming, especially in Europe and Italy. The studies mainly propose three different weed management techniques in the paddy field that can be considered models that can be standardised on farms specialising in their use. These models are the basis of a sustainable fight against ecological and environmental problems thanks to the strict avoidance of chemicals such as herbicides. Value is given to methodologies whose effectiveness has been widely tested. The green mulching technique uses cover crops that effectively control weeds thanks to four main mechanisms. The first consists of the partial inhibition of weeds' germination thanks to competition for water, nutrients, and shading, which are phenomena caused by the presence of cover crops. The second consists of the mulching effect when the crops are chopped or placed on the ground with rice sowing. The third is represented by the phenomena of allelopathy that arise between cover crops and weeds (this aspect will be further explored in Chapter II, https://doi.org/10.3390/plants9030324, https://doi.org/10.1007/s11356-020-10140-4 ). At the same time, the fourth mechanism is linked to the accumulation of phytotoxic compounds following the fermentation caused by the submersion water of the paddy field: the anaerobic environment that is created leads to the formation of organic acids such as acetic acid, butyric and propionic acid. Allelopathy is an interesting aspect offered by plants in this particular agrotechnical. Allelopathy generally produces and releases secondary metabolites, generating inhibitory effects against nearby plant species. The germination, growth and reproduction of target plants can thus be impaired; these aspects were deepened in Chapter II, starting from the organic farmers' experience. The study aimed to define the inhibitory action of Lolium multiflorum Lam., used as a cover crop before rice sowing against Echinochloa oryzoides (Ard.) Fritsch is one of the main rice weeds. Chapter III focuses on evaluating the environmental impact of organic rice cultivation through LCA considering the production scenarios and the agrotechnics described in Chapter I. The LCA approach was adopted"
NEW RESEARCH ON AGROTECHNICS FOR ORGANIC RICE PRODUCTION AND THEIR SUSTAINABILITY ASSESSMENT / V.a.r. Vaglia ; supervisori: S.Monaco Ph.D, C. Bertora PhD ; tutor: S. Bocchi ; coordinator: P. A. Bianco. Dipartimento di Scienze e Politiche Ambientali, 2022 Apr 27. 34. ciclo, Anno Accademico 2021.
VIRTUAL ANTHROPOLOGY AND COMMINGLED SKELETAL REMAINS. RESOURCES AND POTENTIAL FOR PERSONAL IDENTIFICATION IN HUMANITARIAN FORENSIC CONTEXTS
Commingled skeletal remains represent a compelling topic in forensic anthropology, especially in view of the recent mass disasters in the Mediterranean Sea related to the migration crisis. With over 1,000 alleged victims, and thousands of commingled skeletal remains, the shipwreck of April, 8th, 2015 stands among these recurrent tragedies. The advancements of imaging techniques and 3D modelling have brought about innovative tools to forensic anthropology and paved the way to novel research avenues for the analysis of skeletal remains and for personal identification both of the living and the dead, as well. Foremost, research has extensively explored the quantification of similarities and differences between 3D models of anatomical structures as a proxy to infer relationships between AM e PM records and between bones of an individual. The general assumption is that the lowest is the distance between two superimposed models, the most likely is the possibility that the models share a common origin, hence they belong to the same individual.This dissertation delves into these innovative approaches utilizing the 3D-3D superimposition of virtual bone models to facilitate the sorting of commingled remains, ultimately seeking to comprehend the potential practical application in real case scenarios. The structure of this work follows the two main steps for the re- association of commingled bones, i.e., the joint-matching and the pair-matching, resulting in four research lines. The 3D models were produced with several acquisition techniques (laser scanning, sterephotogrammetry, segmentation and surface scanning) from bones of the CAL (Collezione Antropologica LABANOF) skeletal collection, from CT images of a hospital database and from the skeletal remains of the April, 18th, 2015 shipwreck.The 3D-3D joint-matching test on atlases and axes provided an exclusionary method and brought into question the validity of this method to re-associate planar joints. The 3D-3D pair-matching of iliac and of pubic bones further demonstrated that the method can be applied to portions of bones, as long as the extent of fragmentation preserves comparable structures. Indeed, the systematic test on the effect of postmortem damage on the pair-matching verified that 3D-3D superimposition is heavily impaired by the incompleteness of the remains. This study therefore confirmed with novel evidence the major setback that postmortem damage createsfor any anthropological analysis. The application of the technique to a set of scapulae from the commingled remains of the shipwreck of April 18th, 2015, showed that 3D- 3D superimposition may represent a beneficial tool for the resolution of the commingling.This PhD project investigated unaddressed questions about 3D-3D superimposition applied to commingled remains. Indeed, the aim was not to replace visual and osteometric comparisons but to explore the possible application of an additional tool to be used together with traditional methods to strengthen the results within a multidisciplinary approach. Future research would better enlighten the role of up-to- date virtual techniques in the resolution of commingled assemblages from humanitarian contexts.
VIRTUAL ANTHROPOLOGY AND COMMINGLED SKELETAL REMAINS. RESOURCES AND POTENTIAL FOR PERSONAL IDENTIFICATION IN HUMANITARIAN FORENSIC CONTEXTS / A. Palamenghi ; tutor: D. M. Gibelli ; co-tutor: D. De Angelis ; coordinator: C. Sforza. Dipartimento di Scienze Biomediche per la Salute, 2024 May 28. 36. ciclo, Anno Accademico 2023.
PARADIGMI GEOGRAFICI E IDENTITARI IN ELIAS CANETTI ETHOMAS BERNHARD: UNA PROSPETTIVA COMPARATIVA
This dissertation investigates the works of Thomas Bernhard and Elias Canetti, examining their narratives through an interdisciplinary approach that considers the relationship between geography and literature. The authors, whose personal relationship fluctuated between cordiality and conflict, shared a profound Herkunftskomplex – understood as a troubled relationship with their origins – which significantly influenced their literary engagement with space. The purpose of this study is to investigate how this problematic connection to places tied to their roots shaped the construction of the literary geography within their works, where spatial dynamics function as central narrative elements.The thesis is divided into four chapters. The first chapter outlines the theoretical and methodological framework of the study, emphasizing the significance of the ongoing dialogue between geography and literature in contemporary culture.The second chapter delves into the intricate relationship between Elias Canetti and Thomas Bernhard, tracing their personal and literary affinities and divergences. Their acquaintance, initially based on mutual admiration, gradually evolved into a bitter public rivalry. Despite personal tensions, both recognized each other’s literary value. Notably, Canetti’s suggestion that his novel Die Blendung may have influenced Bernhard’s Frost prompts a close examination of the interaction between their poetics.Consequently, the third chapter shows how the concept of Herkunftskomplex is consistently and systematically developed throughout the work of both authors, albeit in distinct ways. Canetti reconstructs his fragmented identity through memory within a cosmopolitan context, while Bernhard uses places as repositories of traumatic experiences, thereby reflecting on the identity crisis of postwar Austria.The fourth chapter undertakes a comparative analysis of Canetti’s Die Blendung and Bernhard’s Frost, examining how these paradigmatic works employ spatial dynamics to shape their narratives. Considering Canetti’s hypothesis regarding the influence of his novel on the genesis of Frost, the comparison focuses on the role of space as a structural narrative element capable of both orienting and disorienting the narrative.
PARADIGMI GEOGRAFICI E IDENTITARI IN ELIAS CANETTI ETHOMAS BERNHARD: UNA PROSPETTIVA COMPARATIVA / I. Manenti ; tutor: M. Castellari ; coordinatrice: L. Pinnavaia. Dipartimento di Lingue, Letterature, Culture e Mediazioni, 2025 Apr 02. 37. ciclo, Anno Accademico 2023/2024.
NEUROINFLAMMATION ACROSS BOUNDARIES: INSIGHTS FROM HUMAN ISCHEMIC STROKE AND BIOMATERIAL CRANIOPLASTY
Neuroinflammation is a complex process, involving cellular and molecular events, which is activated in the central nervous system by acute and chronic insults such as ischemic stroke and traumatic brain injury (TBI). While this response can initially protect against tissue damage and promote repair, its chronic or dysregulated activation often exacerbates neural injury and impairs functional recovery. Glial cells—microglia, astrocytes, and oligodendrocytes—are key players in these mechanisms, influencing both tissue repair and degeneration. In clinical practice, interventions such as cranioplasty (CP) add further complexity, as the introduction of biomaterials and the risk of post-surgical infection can directly and indirectly modulate the inflammatory environment and patient outcomes. On these bases, the overarching aim of this thesis is to elucidate the role of neuroinflammation in two major clinical contexts: (1) human ischemic stroke, with a focus on glial cell responses and fluid biomarker analysis, and (2) CP with biomaterials, particularly in relation to infection risk and strategies for clinical management. To this end, the work is organized in two sections. The first part combines advanced histopathological and molecular analyses to dissect the spatial and temporal dynamics of glial activation and tissue repair in human post-mortem stroke specimens. A particular focus is given to microglial and astrocytic responses in the peri-infarct region, the accumulation and fate of oligodendrocyte precursor cells, and sex-specific differences in glial reactivity. Bulk transcriptomics analysis provides molecular context, revealing robust upregulation of TREM2-driven phagocytic pathways, providing mechanistic context to histological findings. In parallel, fluid biomarker analysis demonstrates that glia-associated markers (sTNFR2 and sTREM2) are increased in the cerebrospinal fluid (CSF) of female stroke patients, while these changes are less pronounced in plasma, indicating a compartment-specific immune response. Ongoing spatial transcriptomics analyses will further refine the understanding of region-specific gene expression and glial dynamics in stroke lesions. Post-mortem human stroke brain specimens, plasma and CSF were made available through a collaboration with prof. Kate Lykke Lambertsen from the University of Southern Denmark (Odense, DK). The second part addresses the management of patients undergoing CP with porous hydroxyapatite implants (CustomBone Service, FinCeramica Faenza Spa, Faenza, Italy), using data from a multicenter cohort and experimental in vitro assays. This work identifies key clinical and surgical predictors of post-CP infection and demonstrates that tailored infection control strategies—including antibiotic regimens and surgical debridement—can increase implant retention and facilitate patient recovery. In vitro, hydroxyapatite exhibits antibacterial properties, supporting its role as a bioactive material that may contribute to a tissue environment more permissive to brain repair, likely indirectly modulating neuroinflammation by reducing infectious complications.In summary, this thesis advances understanding of neuroinflammatory mechanisms and their clinical implications in both spontaneous (ischemic) and post-surgical brain injury. The findings highlight the value of integrating post-mortem human tissue and fluid biomarkers with clinical data to refine patient monitoring and personalized management and support the pursuit of targeted therapies aimed at optimizing neural recovery and outcomes after stroke and neurosurgical intervention.
NEUROINFLAMMATION ACROSS BOUNDARIES: INSIGHTS FROM HUMAN ISCHEMIC STROKE AND BIOMATERIAL CRANIOPLASTY / F.c. Mannella ; tutor: G. D. Norata ; co-tutor: M. Fumagalli ; coordinatore: G. D. Norata. Dipartimento di Scienze Farmacologiche e Biomolecolari Rodolfo Paoletti, 2025. 38. ciclo, Anno Accademico 2024/2025.
BASIC SCIENCE FOR TISSUE ENGINEERING IN VETERINARY ORTHOPAEDIC RESEARCH
The complex micro- and macro-structure of the meniscus requires a multidisciplinary approach to increase the knowledge currently available about the biology that characterizes its development. In the tissue of adult animals, it is possible to distinguish three zones according to the vascularisation, to the biochemical composition and to the regenerative abilities: 1) an outer zone, completely vascularised, with a fibrous-like matrix and capable of regenerating itself; 2) an intermediate zone with transitional characteristics; 3) an inner zone, non-vascularized, with a cartilaginous-like matrix and with poor repair capacity. Therefore, the meniscal tissue engineering, linking the structure and function of the tissues under normal conditions, seeks to develop a compound able to repair injuries of the inner zone, restoring the physiological characteristics to the tissue and avoiding degenerative phenomena. In particular, the aim of this PhD project was to analyse selected endogenous and exogenous factors that can influence the differentiation of meniscal cells. To do this, the project was divided into four parts: 1) role of hypoxia on chondrogenic differentiation of immature meniscal cells; 2) role of hypoxia on vascular factors involved in meniscus maturation; 3) role of endostatin on the chondrogenic differentiation of meniscus cells in a three-dimensional construct; and 4) characterization and validation of micro-computed tomography for the visualization of complex micro-structures such as the meniscal vascular network. Data were obtained through morphological, biochemical, biomechanical, molecular analyses and high-resolution imaging techniques.
BASIC SCIENCE FOR TISSUE ENGINEERING IN VETERINARY ORTHOPAEDIC RESEARCH / V.r. Herrera Millar ; tutor: A. Di Giancamillo ; coordinatore: F. Ceciliani. Dipartimento di Scienze Biomediche per la Salute, 2023 Jul 13. 35. ciclo, Anno Accademico 2022.
STRUCTURE-BASED DESIGN OF GLYCOMIMETIC LIGANDS FOR THE N-TERMINAL DOMAIN OF BC2L-C LECTIN
The prevalence of drug-resistant infections has challenged the existing treatment regimen using antibiotics. There is a need to discover and employ alternative and complementary therapies to counteract these life threatening infections. In fast few decades, the use of anti-adhesion molecules targeting virulence factors such as lectins has been proven an attractive approach to counteract the infections by disarming the pathogens. This thesis work aimed to design glycomimetic antagonists of the N-terminal domain of the BC2L-C lectin (BC2L-C-nt) from the drug-resistant pathogen known as B. cenocepacia. We employed a fragment-based approach to design glycomimetic antagonists of the target protein (BC2L-C-nt). The initial studies were focused towards the binding site prediction and target evaluation by computational tools which identified additional druggable regions near the fucoside binding site in BC2L-C-nt. These additional regions have been explored further to evaluate the druggability by employing virtual screening of a small fragment library. This identified an interesting region (region ‘X’) that could host the drug-like fragments by establishing some key interactions. These interactions with the lectin have been confirmed using a group of biophysical techniques, including X-ray crystallography. Remarkably, the binding mode of one of the fragment (KL3) has been validated by X-ray crystallography at high resolution confirming the ability of site X to host drug-like fragments. Further, the fragments have been chemically connected to the fucose core to obtain high-affinity bifunctional glycomimetic ligands. Interestingly, the crystal complexes of BC2L-C-nt with two bifunctional glycomimetic ligands again confirmed the druggability of the identified site, thus also validated the computational predictions. Hence, the first generation of glycomimetic ligands with binding affinities in micromolar range have been successfully designed. These glycomimetics provide further opportunities to design high-affinity ligands. Future studies based on structure-based approaches and robust synthetic routes to synthesize glycomimetics can lead towards the high-affinity ligands as anti-adhesive agents against B. cenocepacia.
STRUCTURE-BASED DESIGN OF GLYCOMIMETIC LIGANDS FOR THE N-TERMINAL DOMAIN OF BC2L-C LECTIN / K. Lal ; tutor: A. Bernardi, L. Belvisi, A. Imberty, A. Varrot ; coordinatore: E. Licandro. Dipartimento di Chimica, 2021 Dec 15. 33. ciclo, Anno Accademico 2020. [10.13130/lal-kanhaya_phd2021-12-15].
NON - RELIGIOUS DISCOURSES AND THE SECULARIZATION OF THE SECULAR: A COMPUTATIONAL STUDY OF ORGANIZED NON - RELIGION IN THE UNITED STATES AND THE UNITED KINGDOM (1881-2019).
Despite the growth of Non-Religion Studies in recent years, historical changes within secularity remain underexplored and there is a shortage of extensive longitudinal studies dealing with the phenomenon. This shortcoming arguably stems, on one side, from the lack of sufficient granularity in the commonly used longitudinal survey data and, on the other, from a chronic indifference of secularization theory toward the changes occurring within secularity itself. It was argued that secularization is a process of broader (non-)religious change, and that secularization theory can and should be extended to the analysis of non-religion. This thesis employs a series of computational methods, ranging from Topic Modeling to Sentiment Analysis and Dynamic Word Embedding, to explore a large collection of magazines published by two American and two British non-religious organizations between 1881 and 2019. The analysis reveals that the historical fluctuations in the relevance attributed to religion by militant non-religious organizations closely follows fluctuations in the public visibility of religion. Furthermore, in line with the theoretical expectations, the relevance of these topics is consistently higher among groups with a more radical orientation (i.e., among atheists rather than humanists) and in contexts characterized by a lower degree of secularization (i.e., in the US rather than the UK). On the contrary, the demand for positive non-religious identities remained prevalently stable over time showing signs of a descending trend only throughout the latest decades. Sentiment Analysis, in combination with linear regression, confirmed that the proportion of negative sentiment in religion-related topics is positively associated with the perceived relevance of religion. Finally, Dynamic Word Embedding revealed two prevalent shifts. On one side, the critique of religion progressively moved away from an ideological confrontation of religious others and towards a more moderate focus on pragmatic concerns. Second, the conceptualizations of atheism and humanism increasingly converged, emphasizing a positive non-religious identity, political activism, and the organizational aspects of the secular movement.
NON-RELIGIOUS DISCOURSES AND THE SECULARIZATION OF THE SECULAR: A COMPUTATIONAL STUDY OF ORGANIZED NON-RELIGION IN THE UNITED STATES AND THE UNITED KINGDOM (1881-2019) / D. Balazka ; supervisor: F. Biolcati Rinaldi, D. Houtman ; co-supervisor: A. Ferrara, G. Watts ; coordinatore: P. Rebughini. - Milano. Università degli Studi di Milano, 2025 Feb 03. 36. ciclo
Let S be a quasi-compact, quasi-separated scheme, such as the spectrum of a field k, and let SH(S) be Morel and Voevodski's stable motivic homotopy category over S. Let R be a motivic ring spectrum in SH(S); we define the motivic Hochschild homology of R via the derived tensor product: MHH(R)=R ∧_{R ∧ R^{op}} R.This spectrum can be considered the motivic analogue of topological Hochschild homology.The purpose of the first part of this thesis is to investigate the homotopy structure of motivic Hochschild homology of MZ/p- the Suslin-Voevodsky mod-p motivic cohomology ring spectrum for p any prime number. We will in particular consider the case S=Spec(k) is the spectrum of an algebraically closed field k of characteristic different from p. In this context, the homotopy groups of M Z/p are a polynomial ring over the field with p elements on a single generator τ in bidegree (0,-1). To do so, we introduce a tri-graded spectral sequence in motivic homotopy groups inspired by an analogue in stable homotopy theory defined by J. Greenlees in a 2016 paper. We will use our spectral sequence to investigate different spectra.As a motivation for the following computations, we first have a look at MHH(MZ/p) itself. Here we produce a first quadrant, multiplicative, tri-graded spectral sequence with E^2_{s,t,*}=π_{s,*}(MHH(MZ/p)) ⊗_{π_{0,*}(MZ/p)} π_{t,*}(A(p)) converging to π_{s+t,*}(MZ/p). A(p) here denotes the dual motivic Steenrod algebra. Notice that each page in our spectral sequences is tri-graded, meaning we can decompose the associated graded object as a direct sum of modules indexed on three natural numbers- the degrees. In this case, we will not provide a direct argument computing all the homotopy groups: we will simply use the spectral sequence to directly determine the structure in low degrees at the prime p=2. This allows us to perceive how abundant are elements of τ-torsion and motivates the following step: study a τ-inverted and mod-τ^{p-1} versions of MHH(MZ/p), which we are abe to determine via analogous spectral sequences.This strategy proves effective: by combining these two computations with some additional algebraic results, it is possible to reconstruct π_{*,*}(MHH(MZ/p)), as proven in a recent paper by Dundas, Hill, Ormsby and Oestvaer.The final part (chapter D) begins with the following observation: in the event the motivic ring spectrum R is E_∞, one has an equivalence MHH(R)=S^{1}_s ⊗ R, where the tensor symbol encodes an action of simplicial sets on Alg_{E_∞}(SH(S)) given by geometric realization along the associated simplicial object. This equivalence is proven as its classical analogue in topology via the equivalences using the bar complex resolution. In this part of the thesis, we extend the above action along the embedding of simplicial sets into motivic spaces, so that we are entitled to tensoring with more exotic motivic spheres, such as the multiplicative group scheme or the projective line.To do so, we tackle the problem from the more general perspective of left tensored ∞-categories. The construction of the extended action moreover allows us to prove that in the event R= R ∧ R is an idempotent E_∞ motivic ring spectrum, such as rational motivic cohomology, we have equivalences: S^1_s ⊗ R=G_m ⊗ R=P^1 ⊗ R=R.Chapters B, C and D already appeared on the arXiv.
MOTIVIC HOCHSCHILD HOMOLOGY / F. Mocchetti ; supervisor: P. A. Oestvaer, M. Spitzweck ; coordinatori: D. P. Bambusi, G. Ciraolo. Dipartimento di Matematica Federigo Enriques, 2025 Feb 10. 37. ciclo, Anno Accademico 2024/2025.
CHARACTERIZATION OF BIOLOGICALLY ACTIVE MOLECULES OFPHARMACOLOGICAL INTEREST EXTRACTED FROM MICROALGAE BYBIOSUSTAINABLE PROCESSES
Microalgae represent a promising source of high-value secondary metabolites, such as antimicrobial, antioxidant, and immunomodulatory compounds, which are of considerable interest to the pharmaceutical industry. However, conventional extraction technologies—often based on environmentally harmful organic solvents—limit their potential for large-scale industrial exploitation.In this context, the research carried out during my PhD program aimed to develop integrated strategies for the sustainable extraction of microalgal bioactive molecules, prioritizing the use of green solvents such as dimethyl carbonate (DMC), in line with the principles of green chemistry.The study focused on the cultivation of the Micractinium pusillum (CELP1) strain, isolated from agricultural water sources in Sicily (Italy), and its comparison with the model species Chlamydomonas reinhardtii cc-1374. Both strains were evaluated in terms of growth kinetics, biomass productivity, extraction yield, and antimicrobial activity, revealing significant differences in their metabolic composition and bioactivity profiles.In parallel, the industrial strain Chlorella sorokiniana was cultivated under both autotrophic and mixotrophic conditions using agro-industrial by-products as alternative nutrient sources, thereby promoting circular bioeconomy models and reducing cultivation costs. The biomass of Chlorella sorokiniana was then subjected to a controlled alkaline treatment with sodium hydroxide (NaOH) to selectively remove chlorophyll, enhancing the purity and quality of the extracts without compromising the integrity of the bioactive compounds.Extracts from Chlorella sorokiniana were characterized by gas chromatography–mass spectrometry (GC–MS) and tested for antimicrobial activity against selected bacterial strains. The results confirmed the effectiveness of green solvent-based extraction methods in preserving and enhancing the bioactive content of microalgal biomass, achieving a significant improvement in antimicrobial properties compared to conventional methods based on chlorinated solvents.
CHARACTERIZATION OF BIOLOGICALLY ACTIVE MOLECULES OFPHARMACOLOGICAL INTEREST EXTRACTED FROM MICROALGAE BYBIOSUSTAINABLE PROCESSES / E. Lio ; supervisore: G. Aldini ; co-supervisore: F. Secundo ; coordinator: G. Vistoli. - Milano. Dipartimento di Scienze Farmaceutiche, 2025 May 09. 37. ciclo
UNE EXPERIENCE COMMUNALE. SOCIETE ET CULTURE POLITIQUE A MARSEILLE ENTRE XIIE ET XIIIE SIECLE.
Until recently, the historiography of medieval communes has struggled to assert itself outside the analytical framework of central and northern Italy of the twelfth and thirteenth centuries. Yet during these two centuries, urban Provence provided a geographical space and a cultural horizon that were particularly conducive to a full development and a conscious use of the political institutions and social languages specific to medieval communal civilisation. Based on the case study of Marseilles, the most important Provençal city of the medieval period and one of the most dynamic Mediterranean ports of the thirteenth century, the aim of this research is to examine once again the question of the socio-political structures of the medieval communes, placing at the centre of the approach the study of city society itself as well as that of the political culture that this society promoted. In contrast to the old interpretative tradition which sought to reduce the importance of towns in the contextof the French Middle Ages, in favour of the prestige accorded to the formation of the central State, a new approach to the Marseilles « communal experience » is thus proposed, reassessing in particular the original political culture that underpinned this experience. This underlines the importance of socio-institutional and socio-political dynamics inunderstanding the nature, locals adaptation and historical evolution of the cultural matrix of the medieval communes.
"UNE EXPERIENCE COMMUNALE. SOCIETE ET CULTURE POLITIQUE A MARSEILLE ENTRE XIIE ET XIIIE SIECLE / G. Ziggiotto ; rapporteur (PR, Université Versailles Saint-Quentin) : Pierre CHASTANG rapporteur (Pr. Ordinario, Università degli Studi di Cagliari)Lorenzo TANZINI examinatrice (Pr. Associato, Università degli Studi di Pisa)Alma POLONI examinateur (MCF, Aix-Marseille Université)François OTCHAKOVSKY-LAURENS examinateur (Pr. Associato, Università degli studi di Verona)Simone BALOSSINO président du jury (PR, Université Paris I Panthéon Sorbonne)Florian MAZEL tutor principale (PR, Aix-Marseille Université)Laure VERDON tutor secondario (Pr. Ordinario, Università degli studi di Milano)Paolo GRILLO. - Aix-en-Provence (MMSH, salle G. Duby). Dipartimento di Studi Storici ""Federico Chabod"", 2024 Sep 14. 36. ciclo, Anno Accademico 2023/2024."
BAYESIAN INFERENCE ON NUCLEAR DATA AND NEUTRON STAR OBSERVATIONS FOR THE NUCLEAR EQUATION OF STATE
Nuclear matter is defined as a uniform infinite system composed of neutrons and protons at zero temperature interacting only through the nuclear force. Although an idealisation, it is achieved, to a good extent, in the interior of nuclei, where the density is roughly constant and the interaction is dominated by the nuclear force, and in neutron stars, where the high densities in their interior exceed the nuclear ones. Finite nuclei deliver information only on symmetric or quasi-symmetric matter, i.e., when the proton and neutron densities are similar; neutron stars deliver information on highly asymmetric matter instead, since they are mostly composed of neutrons. Hence, low-energy nuclear physics has the potential of imposing tight constraints into the physics of the star, and in turn neutron star observations can inform us on the behaviour of nuclear matter at densities unachievable here on Earth.Recently, two nuclear physics observables that are thought to be intimately connected to the physics of a neutron star have challenged nuclear theory. These observables are the nuclear electric dipole polarizability –in a sense analogous to the tidal deformability in a star– and the parity violating asymmetry –that is very sensitive to the pressure felt among neutrons in the surface of the atomic nucleus. No model to date is able to describe these two observables (within 1 standard deviation).The aim of this project is to study the nuclear Equation of State (EoS) using a the EDF formalism that properly describe the above mentioned observables and that includes statistically sound theoretical errors, and then bridge this information to the study of neutron stars.Within the standard Skyrme functional ansatz, we build a reliable probability distribution for a combination of nuclear matter parameters (NMP) and Skyrme parameters (which are needed to constrain all the terms of the functional) using a combined Bayesian inference of a large set of EoS-sensitive nuclear structure data. Beyond the usual ground state properties like binding energies and charge radii, we also included the above mentioned polarizabilities and parity-violating asymmetries of 208Pb and 48Ca.The result is a multivariate probability distribution for the NMPs and Skyrme parameters.One of the most delicate aspects of the analysis was handling the uncertainties associated with each measurement, as Skyrme energy density functionals cannot always reproduce experimental data within experimental errors, particularly for binding energies.This led us to discover an interplay between the binding energies and the polarizabilities of 208Pb and 48Ca, which favours a soft nuclear equation of state.Furthermore, the posterior distribution can be used as a prior distribution in a successive Bayesian analysis, this time using astrophysical observations as constraints. This way, this second posterior distribution of NMPs will be informed by both nuclear physics and astrophysics. We will show that the constraints from nuclear experiments are well compatible with the theoretical predictions for infinite pure neutron matter from ab initio modelling, and those constraints additionally indicate the existence of interesting structures in the EoS of neutron stars. We will discuss the final predictions on some selected static properties of neutron stars, which can be computed from the distribution of NMPs. We will devote further attention to the composition of the star crust, which is computed consistently with the star EoS within the extended Thomas-Fermi formalism.
BAYESIAN INFERENCE ON NUCLEAR DATA AND NEUTRON STAR OBSERVATIONS FOR THE NUCLEAR EQUATION OF STATE / P. Klausner ; direttore di tesi: G. Colò, F. Gulminelli ; coordinatori: A. Mennella, A. Viveut. Dipartimento di Fisica Aldo Pontremoli, 2025 Dec 09. 38. ciclo, Anno Accademico 2024/2025.
LA SUPERFICIE DI MEZZO. L¿ECOSISTEMA DELLE INTERFACCE DIGITALI: UNA TASSONOMIA CRITICA
Although part of our daily lives for decades, interfaces are a hard to define technology. Whileas many different disciplines and approaches have tried to give a definition of it, little effort has been done to classify, distinguish and define different categories of interfaces. The present work aims to investigate different definitions and mechanisms pertaining to the concept of interface through philosophy, media archaeology and disability and critical studies to structure a critical taxonomy of the computer interface. The grand set computer interfaces constitute has been divided in three main ontological classes, each associated with a strong norm that defines all other categories in the same class. Furthermore, the work proposes to observe how the normalizing mechanisms and the processes of exclusion, descending from different devices and previous technology, contribute to the definition of the norm itself on the bodily, cultural, social and political levels.
LA SUPERFICIE DI MEZZO. L' ECOSISTEMA DELLE INTERFACCE DIGITALI: UNA TASSONOMIA CRITICA / A. Costella ; tutor: A. Pinotti ; cotutor: A. Ferrara ; coordinatore: N. Guicciardini. Dipartimento di Filosofia Piero Martinetti, 2025. 37. ciclo, Anno Accademico 2024/2025.
INVOLVEMENT OF THE CONTACT PATHWAY IN COVID-19 COAGULOPATHY
Background. A novel acquired coagulopathy characterized by a severe procoagulant imbalance is common in COVID-19 patients and is associated with the clinical severity of the disease.Aims. Our study aims at elucidating the underlying mechanisms of coagulation activation in COVID- 19 patients.Methods. The study is composed of two phases. In the first and second part, 62 and 112 symptomatic COVID-19 patients, respectively, were consecutively enrolled and stratified into 3 groups based on the intensity of care: low, requiring only high-flow oxygen by nasal cannula (21 and 23 patients); intermediate, requiring continuous positive airway pressure (20 and 47 patients); high, requiring mechanical ventilation (20 and 42 patients). During the first phase of the study blood samples were tested for markers currently used to diagnose disseminated intravascular coagulation (DIC) (PT, aPTT, fibrinogen, D-dimer, platelet count), the pro- (FII, FVIII) and anticoagulant factors (antithrombin, protein C, protein S) and those indicating endothelial perturbation (von Willebrand factor). During the second phase of the study blood samples were tested for markers of activation of the intrinsic pathway (FXI, FXII, FXIa, FXIIa) together with its physiologic inhibitor (C1-inhibitor), of the extrinsic pathway (FVII, FVIIa), global activation of the coagulation cascade (D-dimer, FDP, FM) and fibrinolysis (plasminogen, t-PA, 𝛼2-antiplasmin, PAI-1).Results. Results of the first phase of the study demonstrated that COVID-19 coagulopathy represents a new entity, not fulfilling the criteria for DIC, characterized by endothelial activation, while the second part of the study showed a prevalent activation of the contact pathway over the extrinsic pathway of the coagulation cascade.Conclusions. Our study showed an acquired coagulopathy associated with hyperacute inflammation, hypercoagulability, and endothelial perturbation proportional to the clinical severity3of the infection, characterized by a prevalent activation of the intrinsic pathway of the coagulation cascade, opening the possibility for targeted therapies.
INVOLVEMENT OF THE CONTACT PATHWAY IN COVID-19 COAGULOPATHY / M. Capecchi ; tutor: F. Payvandi ; coordinator: C. Sforza. Università degli Studi di Milano, 2022 Jun 07. 34. ciclo, Anno Accademico 2021.
STRUCTURAL INVESTIGATION OF EXTRACELLULAR VESICLES AND THEIR INTERACTION WITH BIOMIMETIC INTERFACES
ABDALLA, SALLY MAHMOUD HELMY MOHAMED ABDELHAMID
Over the last decade, research about extracellular vesicles (EVs), the nanosized membrane bound vesicles, secreted by all kinds of cells to deliver a physiological cargo and their influence on the cell-mediated communication has been rising as a hot topic. Different EVs, have different characteristics and activities, so their assessment will allow for their engineering and development for medical purposes. According to that, researchers have been applying mainly biological and biochemical approaches for studying EVs’ properties, including their common biomarkers, protein, and lipid profiles, to exploit them for therapeutic or diagnostic purposes. Here, an alternative novel physical approach to study EVs’ properties is carried by inspecting the EVs’ physical characteristics and connecting them to their activity. In the current study, multiple complementary physical methods are applied to study the characteristics of a new isolation of EVs from neuronal cells (N2a cells) after being treated with a promising Parkinson’s Disease drug molecule, namely the oligosaccharide OligoGM1 (hence, named as Oligo EVs). Those Oligo EVs (that are EVs purified from cells treated with OligoGM1) were studied in comparison with the Ctrl EVs (isolated from the untreated cells). Dynamic light scattering (DLS), nanoparticle tracking analysis (NTA) and atomic force microscopy (AFM) were applied for studying the difference between the EVs’ hydrodynamic and geometric diameters. Additionally, the EVs’ spectroscopic molecular profiles and protein/lipid ratios were investigated by attenuated-total-reflection Fourier-transform infrared spectroscopy (ATR-FTIR), in addition to performing a structural study of their surrounding membrane, using small-angle X-ray scattering (SAXS). Each applied technique has provided particular information of the subtle differences between the characteristics of both types of EVs under study. Then, an investigation of the EVs uptake mechanisms was carried with bottom-up approaches. In this contest, there is an urging need to design different membrane models with different complexities and geometries, to be used as biomimetic interfaces for mimicking the interaction mechanisms of incoming macromolecules such as the EVs. Indeed, the design of such model membranes can be considered as technique-specific. The requirements of each applied physical approach entail specific parameters to be tuned in the designed model membrane, such as its transition temperature, deuteration degree, bilayer thickness, transverse, and lateral structure. Therefore, in parallel, an original study was conducted on a novel model membrane made of the phospholipid 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), sphingomyelin (SM) and cholesterol (Chol), according to the molar ratio of DMPC:SM:Chol; 2:1:0.15, to mimic the structure of the lipid rafts (i.e. phase separated highly ordered lipid domains within a bulk fluid phase lipid), to be used as a biomimetic interface for studying the uptake mechanisms of the EVs. DMPC was used as the main matrix for this model in contrast to other common lipids of lower transition temperatures (Tm) as 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), due to its better accessibility by a wider range of physical techniques such as differential scanning calorimetry (DSC) because of its higher Tm, and neutron reflectometry (NR) by benefiting from its commercially available deuterated forms. Hence, after building-up the lipid raft model, DSC, NR and AFM, were used in a complementary fashion to fully characterize that model membrane thermotropically, as well as its transverse/lateral structure at different temperatures following the organisation and formation of its phase separated domains upon lipid melting. After setting up the different model membranes to be used, the interaction and uptake mechanisms of the EVs were thoroughly studied where many details were unveiled. The pr
STRUCTURAL INVESTIGATION OF EXTRACELLULAR VESICLES AND THEIR INTERACTION WITH BIOMIMETIC INTERFACES / S.m.h.m.a. Abdalla ; main supervisor: V. M. Rondelli ; co-supervisor: P. Brocca ; external co-supervisor: L. Casalis ; director of the thesis: R. Vecchi. - Dipartimento di Fisica, UNIMI. Dipartimento di Biotecnologie Mediche e Medicina Traslazionale, 2024 Jul 16. 36. ciclo, Anno Accademico 2023/2024.
WATER BUFFALO MICROBIOTA AND IMMUNITY DURING INFECTIOUS DISEASES
The thesis aims to investigate the immune response of water buffaloes (Bubalus bubalis) during infectious diseases (e.g. mastitis, brucellosis and tuberculosis) and productions diseases, such as peripartum related metabolic diseases or stress. Given the relationship between the microbial environment and the immune system, the microbiota content of milk has been identified as well. The immune system of water buffalo has been poorly addressed so far, not to mention the microbiota, which was unknown. Dairy water buffaloes are sensitive to the same diseases as dairy ruminants, such as for example mastitis, tuberculosis, brucellosis, but the impact of these diseases on water buffaloes’ immune system are unknown. Water buffalo presents anatomical (e.g. at mammary gland and skin level) and physiological (peripartum and infectious diseases) differences as compared to cow and other dairy ruminants. Therefore, it is evident that both the immune system and microbiota could present several differences. In my PhD thesis, I tried to address some of these issues. Firstly, the milk microbiota of water buffaloes was investigated in relation to mastitis disease. Secondly, the evaluation of the immune response, in terms of gene expression ad miRNAs, was carried out in animals affected by brucellosis and tuberculosis. Finally, the characterization of the transition period was performed measuring acute phase proteins to assess the inflammation status during the transition period. The water buffalo milk microbiota was described herein for the first time. The variation in composition and structure was demonstrated depending on the pathological status of the animals. A change in milk microbial community was observed in clinical and subclinical mastitis-affected quarter milk samples as compared with a health condition, as well as a reduction of microbiota diversity in mastitis-affected quarters. The treatment of subclinical mastitis quarters using inactivated cultures of Lactobacillus rhamnosus was performed in order to unravel the effect of alternative strategies to the therapeutic use of antibiotics. A chemotactic response at quarter level was observed, but no change neither in milk microbiota nor in microbiological culture was found after treatment, suggesting that the use of pro- and pre-biotic for mastitis treatment needs to be further investigated. Given that, so far and in cow, each udder quarter has been regarded as an entity not related to the other three, the hypothesis of the presence of interdependence within milk udder quarters was demonstrated, studying the intra- and inter-individual variation in milk microbiota. Individual signatures may explain the higher similarity between milk microbiota of quarters within the same udder, suggesting that the independence of milk quarters has been challenged and needs to be taken into account when performing microbiota studies. Finally, in order to improve the taxonomic resolution of the experimental design, milk microbiota was characterized using a longer amplicon sequencing by using the full-length 16S marker ( ̴ 1500pb) taking advantage of Nanopore technology, that confirmed the higher resolution power as compared with short-read sequencing ( ̴ 250pb) and could provide a supportive role for microbiological culture technique. The investigation of a possible biomarker for brucellosis in vaginal fluid and serum revealed that levels of 10 miRNAs were modulated in vaginal secretion of seropositive compared to seronegative animals. Furthermore, the diagnostic accuracy of the combination of six miRNAs was good with a sensitivity of 95.45 and a specificity of 85, suggesting that a multimiRNAs panel would allow an early and fast procedure for brucellosis diagnosis. A modification in the immune response during tuberculosis disease was observed, aiming to address the main issue of false-positive during routine TB diagnosis. An unbalanced of cytokines expressi
WATER BUFFALO MICROBIOTA AND IMMUNITY DURING INFECTIOUS DISEASES / C. Catozzi ; tutor: F. Ceciliani, A. Sanchez ; phd coordinator: v. Grieco. DIPARTIMENTO DI MEDICINA VETERINARIA, 2020 Feb 06. 32. ciclo, Anno Accademico 2019. [10.13130/catozzi-carlotta_phd2020-02-06].
CHILDREN'S MATTERS: NEGOTIATING ILLNESS IN EVERYDAY INTERACTIONS AT HOME AND SCHOOL IN GHANA
Focusing on sensations of not feeling well or illness complaints that have not yet been given a diagnosis by a health professional, this research asks how children in an adult-centered and pluralistic context of health care negotiate their illness experiences at home and school. The ways in which children actively construct, frame and deal with their illness, and the role their social position (gender and social class) in the Ghanaian context plays in this process is explored. The research highlights how children intentionally navigate relations and interactions with adults, make use of their social and physical environment to contest, self-diagnose, self-medicate and to deal with their sensations of not feeling well.
CHILDREN'S MATTERS: NEGOTIATING ILLNESS IN EVERYDAY INTERACTIONS AT HOME AND SCHOOL IN GHANA / C. Santah ; supervisors: R. Reis, L. M. Leonini ; co-supervisor: C. Broer, R. Bosisio ; director of doctoral program: M. Barisione. Dipartimento di Scienze Sociali e Politiche, 2020 Mar 27. 30. ciclo, Anno Accademico 2017. [10.13130/santah-colette_phd2020-03-27].