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Emerging plant-based proteins: comparison from extraction to functional properties and food application
Get PDFThe increasing demand for alternatives to meat food products, which is linked to ethical and environmental reasons, highlights the necessity of using different protein sources. Plant proteins provide a valid option, thanks to the relative low costs, high availability and wide supply sources. The current process used to produce plant concentrates and isolates is the alkaline extraction followed by isoelectric precipitation. However, despite the high purity of the proteins, it presents some drawbacks. Innovative protein extraction processes are emerging, with the aim of reducing the environmental impact and the costs, as well as improving the functional properties. In this study, the traditional wet protein extraction and another simplified wet process were used to obtain protein-rich extracts out of different plants. The sources considered in the project were de-oiled sunflower and canola, chickpea, lentils, and the camelina meal, an emerging oleaginous seed interesting for its high content of omega 3. The extracts obtained from the two processes were then analysed for their capacities to hold water and fat, to form gel and a stable foam. Results highlighted strong differences concerning the protein content, yield and functionalities. The extracts obtained with the alkaline process confirmed the literature data about the four plant sources (sunflower, canola, chickpea and lentils) and allow to obtain a camelina concentrate with a protein content of 63 % and a protein recovery of 41 %. The second easiest process was not effective to obtain a protein enrichment in oleaginous sources, whereas an enrichment of 10 and 15 % was obtained in chickpea and lentils, respectively. The functional properties were also completely different: the easiest process produced protein ingredients completely water-soluble at pH 7, with a discrete foaming capacity compared to the extracts obtained with alkaline process. These characteristics could make these extracts suitable for the plant milk-analogue products
Mathematical and Numerical Models for Boundary Optimal Control Problems Applied to Fluid-Structure Interaction
Get PDFThe main purpose of this work is to develop mathematical and
numerical methods for the optimal control of fluid-structure interaction simulations. In particular, we focus on the Koiter shell fluid-structure model and on the adjoint formalism for the control problem. Using the Koiter approach, the dimensionality of the solid is reduced to reduce the computational cost of the fluid-structure simulations. In order to couple the fluid and the structure domains, the Koiter shell equations are embedded into the fluid equations as a Robin boundary condition. The coupling fluid-structure conditions are automatically treated in an implicit way, so the stability of the numerical scheme is preserved.
This model has many applications in cases where a fluid interacts with a thin membrane that deforms mainly in the normal direction. Then, an adjoint-based optimal control theory of the presented Koiter fluid-structure model in the steady case is studied. In fact, a boundary optimal control theory is applied to the fluid-structure Koiter model, including the existence of the solution of the fluid-structure problem, the existence of the optimal solution and regularity and differentiability properties.
Moreover, the fractional operators are introduced to be applied to the framework described above, in order to model properly the regularization term in the boundary optimal control problems.
All the numerical simulations presented in this work, with the exception of the fractional simulations, have been simulated with the in-house multigrid finite element based code FEMuS
Innovative methodologies for enhancing the regionalisation of rainfall-runoff model parameters
Get PDFThe present research work focuses on the regionalisation of rainfall-runoff model parameters, fundamental for the implementation of hydrological models in ungauged basins and needed to reproduce the actual sequence of river discharge in time. Regionalisation of parameters is based on the transfer of information from hydrologically similar gauged basins to ungauged basins. This work provides further insights on parameter regionalisation and catchment similarity through the application of innovative methodologies to support the existing knowledge.
The first experiment develops a methodology to test the robustness of regionalisation procedures to the availability of data in the study region. In particular, the effect of the density of streamflow gauging stations and the topological relationships between their corresponding drainage basins on the different regionalisation techniques is investigated. Such work provides useful information for the choice of the most appropriate method, based on data availability in the region.
The focus is then moved to the value of hydrological similarity at sub-basin scale. Driven by the fact that similarity is generally defined between entire catchments, neglecting some significant differences in the within-basin rainfall-runoff transformation processes, a methodology to differentiate hydrological processes at sub-basin scale and to transfer model parameters from similar sub-basins is proposed. The analysis is based on the diversification of the parameter values, and therefore of the corresponding hydrological dynamics, across elevation, one of the main factors influencing the runoff generation processes.
Finally, an innovative catchment signature is proposed for improving our knowledge about hydrological similarity, meant for the delineation of hydrologically similar regions. A new methodology for identifying the dominant rainfall-runoff transformation dynamics is presented: the interaction between the entire time-series of runoff generation forcings and runoff itself is quantified taking advantage of the concepts of the Information Theory and used to characterise catchments with promising results
La valeur contraignante des droits sociaux fondamentaux fixés par le Titre IV de la Charte des droits fondamentaux de l’UE
Get PDFIl dibattito giuridico, politico ed economico in tema di protezione dei diritti sociali nel contesto del processo di integrazione europea risale alle origini stesse di tale processo. A circa dieci anni dall’attribuzione alla Carta dei diritti fondamentali dell’UE dello stesso valore giuridico dei Trattati, sembra possibile tracciare un primo bilancio in termini di aspettative inverate ed aspettative non soddisfatte quanto alla giustiziabilità dei diritti sociali fondamentali contenuti nel Titolo IV direttamente o indirettamente al rapporto di impiego (articoli da 27 a 34). A questo fine, la tesi ha adottato una struttura tripartita capace di coniugare una metodologia di analisi pratica, in relazione alla giurisprudenza della Corte di giustizia UE post Lisbona (Capitolo III), con un’analisi dal carattere maggiormente teorico-ricostruttivo (Capitoli I e II). La ricerca cerca di dimostrare come tanto i più interessanti sviluppi quanto i limiti registratisi nella giurisprudenza della CGUE (rispettivamente, l’efficacia diretta, anche orizzontale, di alcuni di essi negli ordinamenti nazionali, e la determinazione del loro ambito di applicazione e delle modalità di interazione con il diritto secondario e primario) possano essere meglio compresi tenendo in debito conto il percorso ed il significato di ciascuno di tali diritti nel contesto del processo di integrazione europea. In breve, si sostiene che la “costituzionalizzazione” dei diritti sociali in esame abbia certamente un “valore aggiunto” che, tuttavia, riflette la natura sui generis e le finalità dell’Unione, il sistema di riparto di competenze tra Stati membri e Unione, ed il significato ultimo del sistema UE di tutela dei diritti fondamentali. Ciò non toglie che questi caratteri, con il tempo, possano infine mutare.The legal, political, and economic debate on the protection of social rights in the context of the European integration process dates back to the very origins of this process. The Charter of Fundamental Rights of the EU acquired binding character in December 2009, and most notably it acquired the same legal value as the Treaties. Roughly ten years later, it seems possible to carry out a first assessment draw of the promises kept and the promises broken as for the justiciability of the fundamental social rights enshrined in Title IV which directly or indirectly relating to the employment relationship (Articles 27-34). To this end, the thesis is comprised of three parts. This tripartite structure allows combining two research methods, namely a case law analysis, in relation to the post-Lisbon case law of the Court of Justice of the EU (Chapter III), and a historical and theoretical analysis (Chapters I and II). The major goal of the research is to demonstrate that both the most interesting developments in the CJEU’s case law as well as the most significant open issues can be better understood by taking due account the trajectory and the role of each of these rights within the European integration process. In short, it is argued that the ‘constitutionalisation’ of social rights analysed in this thesis undoubtedly has an ‘added value’. The latter, however, reflects the sui generis nature and goals of the Union, its system of competences, and the ultimate function of the EU system of fundamental rights protection. Despite the fact that these characteristics may eventually change over time.Le débat juridique, politique et économique sur la protection des droits sociaux dans le contexte du processus d’intégration européenne remonte aux origines mêmes de ce processus. Une dizaine d’années après l’attribution à la Charte des droits fondamentaux de l’UE de la même valeur juridique que les traités, il semble possible de dresser un premier bilan en termes d’attentes avérées et d’attentes non satisfaites en ce qui concerne la justiciabilité des droits sociaux fondamentaux contenus dans son Titre IV et directement ou indirectement liés à la relation de travail (articles 27 à 34). À cette fin, la thèse a adopté une structure tripartite capable de combiner une méthodologie d’analyse pratique, en rapport avec la jurisprudence de la Cour de justice de l’UE post-Lisbonne (Chapitre III), avec une analyse ayant un caractère plus théorique-reconstructif (Chapitres I et II). La recherche vise à montrer comment autant les développements les plus intéressants que les limites relevées dans la jurisprudence de la CJUE peuvent être mieux compris en prenant dûment compte du parcours et de l’importance de chacun de ces droits dans le contexte du processus d’intégration européenne. En somme, il est soutenu que la “constitutionnalisation” des droits sociaux analysés présente certainement une «valeur ajoutée» qui, toutefois, reflète la nature sui generis et les objectifs de l’Union, son système des compétences, et la fonction du système de protection des droits fondamentaux de l’UE. Ce qui précède n’implique pas que ces caractéristiques ne puissent pas éventuellement changer à l’avenir
Reasonableness in European and Chinese Contract Law. A Comparative Law Study
Get PDFContemporary private law, in teh last few decades, TEMPhas been increasingly characterized by teh spread of general clauses and standards and by teh growing role of interpreters in teh framework of teh sources of law. dis process TEMPhas also consistently effected those systems dat are not typically centered on judge-made law. In particular in contract law general clauses and standards has assumed a leading role and has become protagonists of processes of integration and harmonization of teh law. Wifin dis context, teh reasonableness clause TEMPhas come to teh attention of scholars, emerging as a new element of connection between different legal systems -first of all between common law and civil law – and even between different legal traditions.
dis research aims at reconstructing teh patterns of emersion and evolution of teh TEMPprincipal of reasonableness in contract law both wifin European Union Law and in teh Chinese legal system, in order to identify evolutionary trends, processes of emersion and circulation of legal models and teh scope of operation of teh TEMPprincipal in teh two contexts.
In view of teh increasingly intense economic relations between Europe and China, wifin teh framework of teh new project called Belt and Road Initiative, a comparative survey of dis type can foster mutual understanding and make communications more TEMPeffective, at teh level of legal culture and commercial relations, and to support teh processes of supranational harmonization of contract law rules
Role of the OPA3 protein in the pathogenesis of neurodegenerative diseases
Get PDFOPA3 è una proteina codificata dal genoma nucleare che, grazie a una sequenza di targeting mitocondriale, viene indirizzata ai mitocondri dopo la sua sintesi. Le mutazioni nel gene OPA3 sono associate a due patologie neurodegenerative: la Sindrome di Costeff, causata da mutazioni recessive, e una forma di atrofia ottica dominante che si manifesta con cataratta e spesso sordità.
L’esatta funzione e regolazione della proteina non sono ancora state completamente chiarite, così come la sua localizzazione nella membrana mitocondriale esterna o interna.
Lo scopo di questa tesi era quello di fare luce sulla funzione della proteina OPA3, con particolare interesse alla dinamica mitocondriale e all’autofagia, sulla sua localizzazione subcellulare ed infine di definire il meccanismo patogenetico nelle patologie neurodegenerative causate da mutazioni in questo gene. A questo scopo abbiamo utilizzato sia una linea di neuroblastoma silenziata stabilmente per OPA3 che linee cellulari primarie derivate da pazienti.
I risultati del presente studio dimostrano che la riduzione di OPA3, indotta nelle cellule del neuroblastoma e presente nei fibroblasti derivati dai pazienti, produce alterazioni nel network mitocondriale con uno sbilanciamento a favore della fusione. Questo fenomeno è probabilmente dovuto all’aumento della forma long della proteina OPA1 che è stato riscontrato in entrambi i modelli cellulari. Inoltre, seppur con direzione apparentemente opposta, in entrambi i modelli abbiamo osservato un’alterata regolazione dell’autofagia.
Infine, abbiamo confermato che OPA3 localizza nella membrana mitocondriale interna ed è esposta per gran parte nella matrice. Inoltre, un segnale della proteina è stato trovato anche nelle mitochondrial associated membranes, suggerendo un possibile ruolo di OPA3 nel trasferimento dei lipidi tra i mitocondri e il reticolo endoplasmatico. Abbiamo rilevato un’interazione della proteina OPA3 con l’acido fosfatidico che non era mai stata evidenziata fino ad oggi. Queste osservazioni sono compatibili con le alterazioni della dinamica mitocondriale e la disregolazione dell’autofagia documentate nei modelli studiati.OPA3 is a protein encoded by the nuclear genome and, after its synthesis, is directed to mitochondria by a mitochondrial targeting sequence. Mutations in the OPA3 gene are responsible for two neurodegenerative diseases: Costeff syndrome, caused by recessive mutations, and a form of dominant optic atrophy with cataracts and hearing loss.
To date, the exact function and regulation of the protein have not yet been understood, as well as its localization in the outer or inner mitochondrial membrane.
This thesis aimed to shed light on the subcellular localization and function of the OPA3 protein, with particular focus on mitochondrial dynamics and autophagy, and to define the pathogenic mechanism of OPA3 mutations causing neurodegenerative diseases. For this purpose, we used neuroblastoma cell line stably silenced for OPA3 and patient-derived primary cell lines.
The results of the present study demonstrate that a reduction of OPA3 content, induced in neuroblastoma cells and naturally occurring in patient-derived fibroblasts, produces alterations in the mitochondrial network with an unbalance toward fusion. This phenomenon is probably due to the increase in the long form of the OPA1 protein in both cell models. Furthermore, we observed an altered regulation of autophagy.
Finally, we confirmed that OPA3 localizes in the inner mitochondrial membrane and is largely exposed in the matrix. Furthermore, the presence of OPA3 was also found in mitochondrial associated membranes, suggesting a possible role of the protein in lipid transfer between mitochondria and the endoplasmic reticulum.
We detected an interaction of the OPA3 protein with phosphatidic acid that had never been shown until now. These observations are compatible with the alterations in mitochondrial dynamics and autophagic dysregulation documented in the models examined
Altered transcriptional and epigenetic regulation affects brain cells proliferation and differentiation in the ultra-rare genetic demyelinating disease AGC1 deficiency: a study on in vitro models
Get PDFAGC1 deficiency is a rare demyelinating disease caused by mutations in the SLC25A12 gene, which encodes for the mitochondrial glutamate-aspartate carrier 1 (AGC1/Alarar), highly expressed in the central nervous system. In neurons, impairment in AGC1 activity leads to reduction in N-acetyl-aspartate, the main lipid precursor for myelin synthesis (Profilo et al., 2017); in oligodendrocytes progenitors cells, AGC1 down regulation has been related to early arrest proliferation and premature differentiation (Petralla et al., 2019). Additionally, in vivo AGC1 deficiency models i.e., heterozygous mice for AGC1 knock-out and neurospheres from their subventricular zone, respectively, showed a global decrease in cells proliferation and a switch in neural stem cells (NSCs) commitment, with specific reduction in OPCs number and increase in neural and astrocytic pools (Petralla et al., 2019). Therefore, the present study aims to investigate the transcriptional and epigenetic regulation underlying the alterations observed in OPCs and NSCs biological mechanisms, in either AGC1 deficiency models of Oli-neu cells (murine immortalized oligodendrocytes precursors cells), partially silenced by a shRNA for SLC25A12 gene, and SVZ-derived neurospheres from AGC1+/- mice. Western blot and immunofluorescence analysis revealed significant variations in the expression of transcription factors involved in brain cells’ proliferation and differentiation, in association with altered histone post-translational modifications, as well as histone acetylases (HATs) and deacetylases (HDACs) activity/expression, suggesting an improper transcriptional and epigenetic regulation affecting both AGC1 deficiency in vitro models. Furthermore, given the large role of acetylation in controlling in specific time-windows OPC maturation (Hernandez and Casaccia; 2015), pharmacological HATs/HDACs inhibitions were performed, confirming the involvement of chromatin remodelling enzymes in the altered proliferation and early differentiation observed in the AGC1 deficiency models of siAGC1 Oli-neu cells and AGC1+/- mice-derived neurospheres
Analytics and Methods for Logistic System Design and Operations Management
Get PDFAnalytics is the technology working with the manipulation of data to produce information able to change the world we live every day. Analytics have been largely used within the last decade to cluster people’s behaviour to predict their preferences of items to buy, music to listen, movies to watch and even electoral preference. The most advanced companies succeded in controlling people’s behaviour using analytics. Despite the evidence of the super-power of analytics, they are rarely applied to the big data collected within supply chain systems (i.e. distribution network, storage systems and production plants).
This PhD thesis explores the fourth research paradigm (i.e. the generation of knowledge from data) applied to supply chain system design and operations management. An ontology defining the entities and the metrics of supply chain systems is used to design data structures for data collection in supply chain systems. The consistency of this data is provided by mathematical demonstrations inspired by the factory physics theory.
The availability, quantity and quality of the data within these data structures define different decision patterns. Ten decision patterns are identified, and validated on-field, to address ten different class of design and control problems in the field of supply chain systems research
Novel Semiconducting Materials and Thin Film Technologies for High Energy Radiation Detection
Get PDFNowadays the development of real-time ionizing radiation detection system operating over large areas is crucial. Increasing quest for flexible, portable, low cost and low power consumption sensors pushed the scientific community to look for alternative materials and technologies able to fulfill these new requirements. In this thesis the potentiality of organic semiconductors and metal oxides as material platforms for novel ionizing radiation sensors is demonstrated. In particular, organic semiconductors are human tissue-equivalent and this represents a unique and desirable property for the development of dosimeters to be employed in the medical field. The ionizing radiation sensors described in this thesis have been designed, fabricated and characterized during my PhD research and are realized onto polymeric foils leading to flexible devices operating at low voltages, in ambient condition and able to directly detect X-rays, gamma-rays and protons. Following the study of the properties and of the mechanisms of interaction between the radiation and the active layers of the sensors, several strategies have been adopted to enhance the efficiency of these detectors. X-rays dosimeters based on organic semiconductors have been realized presenting record sensitivity values compared with the state of the art for large area radiation detection. The unprecedentedly reported performance led to the possibility to testing these devices in actual medical environments. Moreover, the proof-of-principle demonstration of a dosimetric detection of proton beams by organic-based sensors is reported. Finally, a new sensing platform based on metal oxides is introduced. Combining the advantages of amorphous high mobility oxide semiconductors with a multilayer dielectric, novel devices have been designed, capable of providing a sensitivity one order of magnitude higher than the one shown by the standard RADFETs. Thanks to their unique properties, these sensors have been integrated with a wireless readout system based on a commercial RFID tag and its assessment is presented
Protective effects of podocyte-expressed decay accelerating factor in focal and segmental glomerulosclerosis
Get PDFKidney glomerulosclerosis commonly progresses to end-stage kidney failure, but pathogenic mechanisms are still poorly understood. Here, we show that podocyte expression of decay-accelerating factor (DAF/CD55), a complement C3 convertase regulator, crucially controls disease in murine models of adriamycin (ADR)-induced focal and segmental glomerulosclerosis (FSGS) and streptozotocin (STZ)-induced diabetic glomerulosclerosis. ADR induces enzymatic cleavage of DAF from podocyte surfaces, leading to complement activation. C3 deficiency or prevention of C3a receptor (C3aR) signaling abrogates disease despite DAF deficiency, confirming complement dependence. Mechanistic studies show that C3a/C3aR ligations on podocytes initiate an autocrine IL-1β/IL-1R1 signaling loop that reduces nephrin expression, causing actin cytoskeleton rearrangement. Uncoupling IL-1β/IL-1R1 signaling prevents disease, providing a causal link. Glomeruli of patients with FSGS lack DAF and stain positive for C3d, and urinary C3a positively correlates with the degree of proteinuria. Together, our data indicate that the development and progression of glomerulosclerosis involve loss of podocyte DAF, triggering local, complement-dependent, IL-1β-induced podocyte injury, potentially identifying new therapeutic targets