Analisi molecolare delle fasi evolutive della micosi fungoide

Lo scopo del progetto triennale del dottorato di ricerca è lo studio delle alterazioni genetiche in un gruppo di pazienti affetti da micosi fungoide ed un gruppo di pazienti affetti da sindrome di Sezary. Dalle biopsie cutanee è stato estratto il DNA e analizzato, comparandolo con DNA sano di riferimento, utilizzando la tecnica array-CGH, allo scopo di identificare la presenza di geni potenzialmente implicati nel processo di oncogenesi. Questa analisi è stata eseguita, per ogni paziente, su biopsie effettuate ad una fase iniziale di malattia e ad una fase di progressione della stessa. Sugli stessi pazienti è stata inoltre eseguita un’analisi miRNA. Si ipotizza che il profilo d’espressione dei miRNA possa infatti dare informazioni utili per predire lo stato di malattia, il decorso clinico, la progressione tumorale e la riposta terapeutica. Questo lavoro è stato poi eseguito su biopsie effettuate in pazienti affetti da sindrome di Sezary che, quando non insorge primitivamente come tale, si può considerare una fase evolutiva della micosi fungoide. La valutazione delle alterazioni genetiche, ed in particolare la correlazione esistente tra duplicazione e delezione genetica e sovra/sottoespressione genetica, è stata possibile attraverso l’interpretazione e la comparazione dei dati ottenuti attraverso le tecniche array-CGH e miRNA. Sono stati comparati i risultati ottenuti per valutare quali fossero le alterazioni cromosomiche riscontrate nei diversi stadi di malattia. L’applicazione dell’array-CGH e della metodica di analisi mi-RNA si sono rivelate molto utili per l’identificazione delle diverse aberrazioni cromosomiche presenti nel genoma dei pazienti affetti da micosi fungoide e sindrome di Sezary, per valutare la prognosi del paziente e per cercare di migliorare o trovare nuove linee terapeutiche per il trattamento delle due patologie. Lo studio di questi profili può rappresentare quindi uno strumento di grande importanza nella classificazione e nella diagnosi dei tumori.

Design and implementation of a testbed for data distribution management

Data Distribution Management (DDM) is a core part of High Level Architecture standard, as its goal is to optimize the resources used by simulation environments to exchange data. It has to filter and match the set of information generated during a simulation, so that each federate, that is a simulation entity, only receives the information it needs. It is important that this is done quickly and to the best in order to get better performances and avoiding the transmission of irrelevant data, otherwise network resources may saturate quickly. The main topic of this thesis is the implementation of a super partes DDM testbed. It evaluates the goodness of DDM approaches, of all kinds. In fact it supports both region and grid based approaches, and it may support other different methods still unknown too. It uses three factors to rank them: execution time, memory and distance from the optimal solution. A prearranged set of instances is already available, but we also allow the creation of instances with user-provided parameters. This is how this thesis is structured. We start introducing what DDM and HLA are and what do they do in details. Then in the first chapter we describe the state of the art, providing an overview of the most well known resolution approaches and the pseudocode of the most interesting ones. The third chapter describes how the testbed we implemented is structured. In the fourth chapter we expose and compare the results we got from the execution of four approaches we have implemented. The result of the work described in this thesis can be downloaded on sourceforge using the following link: https://sourceforge.net/projects/ddmtestbed/. It is licensed under the GNU General Public License version 3.0 (GPLv3).

Innovative Strategies for the Synthesis of Biologically Active Small Molecules

The post genomic era, set the challenge to develop drugs that target an ever-growing list of proteins associated with diseases. However, an increase in the number of drugs approved every year is nowadays still not observed. To overcome this gap, innovative approaches should be applied in drug discovery for target validation, and at the same time organic synthetic chemistry has to find new fruitful strategies to obtain biologically active small molecules not only as therapeutic agents, but also as diagnostic tools to identify possible cellular targets. In this context, in view of the multifactorial mechanistic nature of cancer, new chimeric molecules, which can be either antitumor lead candidates, or valuable chemical tools to study molecular pathways in cancer cells, were developed using a multitarget-directed drug design strategy. According to this approach, the desired hybrid compounds were obtained by combining in a single chemical entity SAHA analogues, targeting histone deacetylases (HDACs), with substituted stilbene or terphenyl derivatives able to block cell cycle, to induce apoptosis and cell differentiation and with Sorafenib derivative, a multikinase inhibitor. The new chimeric derivatives were characterized with respect to their cytotoxic activity and their effects on cell cycle progression on leukemia Bcr-Abl-expressing K562 cell lines, as well as their HDACs inhibition. Preliminary results confirmed that one of the hybrid compounds has the desired chimeric profile. A distinct project was developed in the laboratory of Dr Spring, regarding the synthesis of a diversity-oriented synthesis (DOS) library of macrocyclic peptidomimetics. From a biological point of view, this class of molecules is extremely interesting but underrepresented in drug discovery due to the poor synthetic accessibility. Therefore it represents a valid challenge for DOS to take on. A build/couple/pair (B/C/P) approach provided, in an efficient manner and in few steps, the structural diversity and complexity required for such compounds.

Topological aspects of the human protein interaction network

It is currently widely accepted that the understanding of complex cell functions depends on an integrated network theoretical approach and not on an isolated view of the different molecular agents. Aim of this thesis was the examination of topological properties that mirror known biological aspects by depicting the human protein network with methods from graph- and network theory. The presented network is a partial human interactome of 9222 proteins and 36324 interactions, consisting of single interactions reliably extracted from peer-reviewed scientific publications. In general, one can focus on intra- or intermodular characteristics, where a functional module is defined as "a discrete entity whose function is separable from those of other modules". It is found that the presented human network is also scale-free and hierarchically organised, as shown for yeast networks before. The interactome also exhibits proteins with high betweenness and low connectivity which are biologically analyzed and interpreted here as shuttling proteins between organelles (e.g. ER to Golgi, internal ER protein translocation, peroxisomal import, nuclear pores import/export) for the first time. As an optimisation for finding proteins that connect modules, a new method is developed here based on proteins located between highly clustered regions, rather than regarding highly connected regions. As a proof of principle, the Mediator complex is found in first place, the prime example for a connector complex. Focusing on intramodular aspects, the measurement of k-clique communities discriminates overlapping modules very well. Twenty of the largest identified modules are analysed in detail and annotated to known biological structures (e.g. proteasome, the NFκB-, TGF-β complex). Additionally, two large and highly interconnected modules for signal transducer and transcription factor proteins are revealed, separated by known shuttling proteins. These proteins yield also the highest number of redundant shortcuts (by calculating the skeleton), exhibit the highest numbers of interactions and might constitute highly interconnected but spatially separated rich-clubs either for signal transduction or for transcription factors. This design principle allows manifold regulatory events for signal transduction and enables a high diversity of transcription events in the nucleus by a limited set of proteins. Altogether, biological aspects are mirrored by pure topological features, leading to a new view and to new methods that assist the annotation of proteins to biological functions, structures and subcellular localisations. As the human protein network is one of the most complex networks at all, these results will be fruitful for other fields of network theory and will help understanding complex network functions in general.

Biomedicine and pain

The focus of my research is on contemporary biomedical construction of pain as an object, i.e. the different ways in which pain has been conceptualized and approached as a specific site of investigation in biomedicine. A significant shift in the scientific conception of pain occured in the second half of XXth century. In 1965, Ronald Melzack and Patrick D. Wall propose the Gate Control theory of pain mechanism. This theory denies a fixed and direct relationship between stimulus and pain perception, and emphazises the role played by psychological factors in pain. The IASP utilizes this perspective on the phenomenon, describing pain as “an unpleasant sensory and emotional experience associated to an actual or potential tissue damage or described in the terms of such a damage.” The relationship between pain and damage is pivotal in the definition of pain as a pathological entity. In particular, the biomedical approach to pain appears to be strongly characterized by a dualistic view of its aetiology. Disease conceptions such as “psychogenic pain” and chronic pain are deeply influenced by the ways in which psychological factors have been interpreted as components, or as causes of pain. In the second part of my dissertation, I focus on fibromyalgia, which is emblematic of the problematic acknowledgment of chronic pain as a disease. Even if fibromyalgia is actually treated in Rheumatology, its status as a disease is blurred, mainly because of its complex symptomatology including both physiological manifestations and psychological ones. In the conclusion, I present a scenario of the different ways in which this disease is dealt with in biomedical knowledge, through medical literature, clinical practice, and patients’ accounts. The findings of an ethnographic enquiry in the Rheumatology Division of a local clinic and a visual research on patients’ experiences are analyzed and discussed.

Assessing the functional structure of molecular transporters by EPR spectroscopy

In this thesis, the self-assembled functional structure of a broad range of amphiphilic molecular transporters is studied. By employing paramagnetic probe molecules and ions, continuous-wave and pulse electron paramagnetic resonance spectroscopy reveal information about the local structure of these materials from the perspective of incorporated guest molecules. First, the transport function of human serum albumin for fatty acids is in the focus. As suggested by the crystal structure, the anchor points for the fatty acids are distributed asymmetrically in the protein. In contrast to the crystallographic findings, a remarkably symmetric entry point distribution of the fatty acid binding channels is found, which may facilitate the uptake and release of the guest molecules. Further, the metal binding of 1,2,3-triazole modified star-shaped cholic acid oligomers is studied. These biomimetic molecules are able to include and transport molecules in solvents of different polarity. A pre-arrangement of the triazole groups induces a strong chelate-like binding and close contact between guest molecule and metal ion. In absence of a preordering, each triazole moiety acts as a single entity and the binding affinity for metal ions is strongly decreased. Hydrogels based on N-isopropylacrylamide phase separate from water above a certain temperature. The macroscopic thermal collapse of these hydrogels is utilized as a tool for dynamic nuclear polarization. It is shown that a radical-free hyperpolarized solution can be achieved with a spin-labeled gel as separable matrix. On the nanoscale, these hydrogels form static heterogeneities in both structure and function. Collapsed regions protect the spin probes from a chemical decay while open, water-swollen regions act as catalytic centers. Similarly, thermoresponsive dendronized polymers form structural heterogeneities, which are, however, highly dynamic. At the critical temperature, they trigger the aggregation of the polymer into mesoglobules. The dehydration of these aggregates is a molecularly controlled non-equilibrium process that is facilitated by a hydrophobic dendritic core. Further, a slow heating rate results in a kinetically entrapped non-equilibrium state due to the formation of an impermeable dense polymeric layer at the periphery of the mesoglobule.

"Tra centro e periferia" Istituzioni e processi di industrializzazione nella Sicilia del secondo dopoguerra

In Sicilia lo strumento principale che ha caratterizzato le modalità d’intervento dei poteri pubblici in economia è stato quello della partecipazione diretta o indiretta, quello cioè della Regione che si propone come ente fautore del sostegno allo sviluppo dell’industria e successivamente come soggetto imprenditore. Le formule organizzative attraverso le quali si è concretizzato l’intervento regionale in economia furono la società per azioni a partecipazione regionale, l’azienda autonoma regionale e l’ente pubblico regionale1. La storia dello sviluppo economico siciliano nel secondo dopoguerra conferma come il ricorso allo strumento dell’ente pubblico economico sia stato molto frequente nella realtà locale come, peraltro, anche in altre Regioni d’Italia. Tracce, queste, di una vicenda storica che intuiamo subito avere complesse implicazioni tali da generare la necessità di interrogarsi sul modo nel quale le istituzioni politiche hanno influito sulle dinamiche economiche siciliane nel secondo dopoguerra; per quanto noti e approfonditi siano stati infatti gli elementi caratterizzanti e i percorsi peculiari dello sviluppo economico siciliano, rimangono scarsamente approfonditi il tenore dei rapporti e i nessi politici, istituzionali ed economici tra centro e periferia, in altre parole rimane ancora parzialmente inesplorata quella parte dell’indagine inerente l’evoluzione dei processi di industrializzazione della Sicilia nel secondo dopoguerra attuata parallelamente dalle autorità regionali e dallo Stato attraverso i loro enti e strumenti. È lecito chiedersi quali siano stati i tempi, le modalità, gli ostacoli e gli eventuali risultati delle azioni di pianificazione intraprese dai poteri pubblici centrali e regionali nella prospettiva dello sviluppo economico del territorio; il coordinamento delle azioni di promozione del progresso industriale si presentava in tal senso, sin dall’inizio, come una delle sfide fondamentali per un adeguato e consistente rilancio economico delle aree più arretrate del Mezzogiorno italiano; ecco che lo studio dei provvedimenti legislativi emanati rappresenta un approfondimento indispensabile e obiettivo primario di questo lavoro.

Metagenomic trajectory of gut microbiome in the human lifespan

Co-evolving with the human host, gut microbiota establishes configurations, which vary under the pressure of inflammation, disease, ageing, diet and lifestyle. In order to describe the multi-stability of the microbiome-host relationship, we studied specific tracts of the bacterial trajectory during the human lifespan and we characterized peculiar deviations from the hypothetical development, caused by disease, using molecular techniques, such as phylogenetic microarray and next-generation sequencing. Firstly, we characterized the enterocyte-associated microbiota in breast-fed infants and adults, describing remarkable differences between the two groups of subjects. Successively, we investigated the impact of atopy on the development of the microbiome in Italian childrens, highlithing conspicuous deviations from the child-type microbiota of the Italian controls. To explore variation in the gut microbiota depending on geographical origins, which reflect different lifestyles, we compared the phylogenetic diversity of the intestinal microbiota of the Hadza hunter-gatherers of Tanzania and Italian adults. Additionally, we characterized the aged-type microbiome, describing the changes occurred in the metabolic potential of the gut microbiota of centenarians with respect to younger individuals, as a part of the pathophysiolology of the ageing process. Finally, we evaluated the impact of a probiotics intervention on the intestinal microbiota of elderly people, showing the repair of some age-related dysbioses. These studies contribute to elucidate several aspects of the intestinal microbiome development during the human lifespan, depicting the microbiota as an extremely plastic entity, capable of being reconfigured in response to different environmental factors and/or stressors of endogenous origin.

Identity formation at the beginning of the twenty-first century: Intersubjectivity, art, and medicine in Siri Hustvedt’s works

„Ich bin, weil du bist“ – so lautet eines der Schlüsselzitate in What I Loved, dem 2003 erschienenen dritten Roman der zeitgenössischen amerikanischen Autorin Siri Hustvedt. Die Bedeutung von Beziehung und Interaktion für die Identitätsbildung spielt eine zentrale Rolle nicht nur in diesem Roman, sondern auch in ihrem Gesamtwerk, das vier Romane, ein memoir, drei Essay-Sammlungen und einen Lyrikband umfasst. Hustvedt erforscht die Identität als ein vielschichtiges Produkt bewusster und unbewusster Verknüpfungen innerhalb der sozialen und biologischen Umwelt. Das Bewusstsein wird als eine dialogisch geprägte Entität gezeigt, dessen Identität erst durch die Beziehung auf ein Anderes geformt werden kann. Um dem Mysterium der menschlichen Identitätsfindung nachzuspüren, bedient sich Hustvedt sowohl philosophischer, psychoanalytischer, biologischer als auch kunsttheoretischer Diskurse. In ihren Romanen stellt sich die Frage nach der Erklärung von Identität als komplexe Problematik dar: Ist die Beziehung zu anderen Menschen vor allem durch unsere Entwicklung als Kind und die Nähe zu Bezugspersonen geprägt? In welchem Ausmaß ist das Empfinden von Subjektivität beeinflusst von körperlichen und unbewussten Mechanismen? Inwiefern ist die Wahrnehmung visueller Kunst eine Kooperation zwischen Betrachter und Künstler? rnDiesen und anderen Fragen geht diese Dissertation nach, indem sie Hustvedts Werk als Anlass für eine Analyse intersubjektiver Strukturen der Identität nimmt. Die Intersubjektivitätsphiloso¬phien von Hegel, Buber, Bakhtin, Husserl, und Merleau-Ponty dienen hierbei als Ausgangspunkt für die Interpretation von relationaler Identität in Hustvedts Werken. Die Dissertation konzentriert sich auf Hustvedts Darstellung der Beziehung zwischen Selbst und Anderem in der Photographie und in der Malerei, der Überschreitung von Körpergrenzen in Hysterie und Anorexie sowie der Auswirkung des Verlustes von Bezugspersonen auf die persönliche Identität. Entscheidend für den Hustvedtschen Kunstbegriff ist das Zusammenspiel von Kunstobjekt, Künstler und Betrachter. Die Grenzen zwischen Innerem und Äußeren werden aufgelöst: mal wird der Rezipient Teil des Kunstwerks, mal verschmilzt der Künstler förmlich mit seinem Objekt. Auch hier wird wiederum deutlich, dass Identität nur in Wechselbeziehung und als zwischenmenschliche Kooperation entsteht. Hustvedt betritt durch ihre einzigartige Auseinandersetzung mit den Wechselbeziehungen und fragilen Grenzen zwischen Ich und Umwelt Neuland auf dem Gebiet der literarischen Identitätsforschung, da sie ihr Prinzip des „mixing,“ des unausweichlichen Eindringens fremder Substanz in die eigene Identität, aus dem Blickwinkel dieser verschiedenen Erklärungsansätze beleuchtet. rn

Knowledge Patterns for the Web: extraction, tranformation and reuse

This thesis aims at investigating methods and software architectures for discovering what are the typical and frequently occurring structures used for organizing knowledge in the Web. We identify these structures as Knowledge Patterns (KPs). KP discovery needs to address two main research problems: the heterogeneity of sources, formats and semantics in the Web (i.e., the knowledge soup problem) and the difficulty to draw relevant boundary around data that allows to capture the meaningful knowledge with respect to a certain context (i.e., the knowledge boundary problem). Hence, we introduce two methods that provide different solutions to these two problems by tackling KP discovery from two different perspectives: (i) the transformation of KP-like artifacts to KPs formalized as OWL2 ontologies; (ii) the bottom-up extraction of KPs by analyzing how data are organized in Linked Data. The two methods address the knowledge soup and boundary problems in different ways. The first method provides a solution to the two aforementioned problems that is based on a purely syntactic transformation step of the original source to RDF followed by a refactoring step whose aim is to add semantics to RDF by select meaningful RDF triples. The second method allows to draw boundaries around RDF in Linked Data by analyzing type paths. A type path is a possible route through an RDF that takes into account the types associated to the nodes of a path. Then we present K~ore, a software architecture conceived to be the basis for developing KP discovery systems and designed according to two software architectural styles, i.e, the Component-based and REST. Finally we provide an example of reuse of KP based on Aemoo, an exploratory search tool which exploits KPs for performing entity summarization.

Proteomics-driven approach for the detection of breast cancer biomarkers

Breast cancer (BC) is the most often diagnosed cancer entity of women worldwide. No molecular biomarkers are usable in the clinical routine for the early detection of BC. Proteomics is one of the dynamic tools for the successful examination of changes on the protein level. In this thesis different proteomics-based investigations were performed for the detection of protein and autoantibody biomarkers in serum samples of BC and healthy (CTRL) subjects. First, protein levels of candidates from previous profiling studies were investigated via antibody-microarray platform. Three proteins were found in distinct levels in both groups: secretoglobin family 1D member 1, alpha-2 macroglobulin and inter-alpha-trypsin inhibitor heavy chain family member 4. The second part was dedicated to the de novo exploration of potentially immunogenic tumor antigens (TA’s) with immunoprecipitation and Western immunoblotting followed by identification over mass spectrometry. Autoantibody levels were verified in individual serum profiling via the protein microarray platform. Two autoantibody’ cohorts (anti-Histone 2B and anti-Recoverin) were found in different levels in both groups. The findings of this PhD thesis underline deregulated serum protein and autoantibody levels in the presence of BC. Further investigations are needed to confirm the results in an independent study population.

Doveri e responsabilita' degli amministratori nella crisi dei gruppi di societa'

Il presente studio si propone di individuare i doveri e le responsabilità, di tipo risarcitorio, degli amministratori, in particolare degli amministratori della società che esercita attività di direzione e coordinamento, in una situazione di crisi o insolvenza nel gruppo, anche in un’ottica di “prevenzione”, e, più precisamente, il complesso di regole di corretta gestione societaria e imprenditoriale, con le quali il silenzio della legge fallimentare in tema di gruppi di società non può non confrontarsi. In particolare, si indagherà sulla possibilità di individuare nel nostro ordinamento giuridico, nel momento di emersione della crisi, doveri di comportamento in capo agli organi di governo della società o ente che esercita attività di direzione e coordinamento, al fine di fronteggiare la crisi, evitando il peggioramento della stessa, ovvero per un risanamento anticipato e, quindi, più suscettibile di esito positivo, nella prospettiva di tutela dei soci c.d. esterni e dei creditori delle società figlie e, nello stesso tempo, dei soci della capogruppo medesima e, quindi, in una prospettiva più ampia e articolata rispetto a una società individualmente considerata. L’oggetto dell’analisi viene introdotto mediante un inquadramento generale della disciplina in materia di gruppi di società presente nel nostro sistema normativo, con particolare riguardo alla disciplina dell’attività di direzione e coordinamento introdotta dal legislatore della riforma del diritto societario (d.lgs. 17 gennaio 2003, n. 6) con gli artt. 2497 ss. cod. civ.. Nella seconda parte verranno individuati e approfonditi i criteri e i principi dai quali ricavare le regole di governance nei gruppi di società e la relativa responsabilità degli amministratori nelle situazioni di crisi nel gruppo. Sulla scorta delle suddette argomentazioni, nell'ultima parte verranno individuate le regole di gestione nell'ambito del gruppo nel momento di “emersione” della crisi e, in particolare, i possibili “strumenti” che il nostro legislatore offre per fronteggiarla.

Untersuchungen zur Strahldynamik am Harmonischen Doppelseitigen Mikrotron von MAMI-C

Das Institut für Kernphysik der Universität Mainz betreibt seit 1990 eine weltweit einzigartige Beschleunigeranlage für kern- und teilchenphysikalische Experimente – das Mainzer Mikrotron (MAMI-B). Diese Beschleunigerkaskade besteht aus drei Rennbahn-Mikrotrons (RTMs) mit Hochfrequenzlinearbeschleunigern bei 2.45 GHz, mit denen ein quasi kontinuierlicher Elektronenstrahl von bis zu 100 μA auf 855MeV beschleunigt werden kann.rnrnIm Jahr 1999 wurde die Umsetzung der letzten Ausbaustufe – ein Harmonisches Doppelseitiges Mikrotron (HDSM, MAMI-C) – mit einer Endenergie von 1.5 GeV begonnen. Die Planung erforderte einige mutige Schritte, z.B. Umlenkmagnete mit Feldgradient und ihren daraus resultierenden strahloptischen Eigenschaften, die einen großen Einfluss auf die Longitudinaldynamik des Beschleunigers haben. Dies erforderte die Einführung der „harmonischen“ Betriebsweise mit zwei Frequenzen der zwei Linearbeschleuniger.rnrnViele Maschinenparameter (wie z.B. HF-Amplituden oder -Phasen) wirken direkt auf den Beschleunigungsprozess ein, ihre physikalischen Größen sind indes nicht immer auf einfache Weise messtechnisch zugänglich. Bei einem RTM mit einer verhältnismäßig einfachen und wohldefinierten Strahldynamik ist das im Routinebetrieb unproblematisch, beim HDSM hingegen ist schon allein wegen der größeren Zahl an Parametern die Kenntnis der physikalischen Größen von deutlich größerer Bedeutung. Es gelang im Rahmen dieser Arbeit, geeignete Methoden der Strahldiagnose zu entwickeln, mit denen diese Maschinenparameter überprüft und mit den Planungsvorgaben verglichen werden können.rnrnDa die Anpassung des Maschinenmodells an eine einzelne Phasenmessung aufgrund der unvermeidlichen Messfehler nicht immer eindeutige Ergebnisse liefert, wird eine Form der Tomographie verwendet. Der longitudinale Phasenraum wird dann in Form einer Akzeptanzmessung untersucht. Anschließend kann ein erweitertes Modell an die gewonnene Datenvielfalt angepasst werden, wodurch eine größere Signifikanz der Modellparameter erreicht wird.rnrnDie Ergebnisse dieser Untersuchungen zeigen, dass sich der Beschleuniger als Gesamtsystem im Wesentlichen wie vorhergesagt verhält und eine große Zahl unterschiedlicher Konfigurationen zum Strahlbetrieb möglich sind – im Routinebetrieb wird dies jedoch vermieden und eine bewährte Konfiguration für die meisten Situationen eingesetzt. Das führt zu einer guten Reproduzierbarkeit z.B. der Endenergie oder des Spinpolarisationswinkels an den Experimentierplätzen.rnrnDie Erkenntnisse aus diesen Untersuchungen wurden teilweise automatisiert, so dass nun den Operateuren zusätzliche und hilfreiche Diagnose zur Verfügung steht, mit denen der Maschinenbetrieb noch zuverlässiger durchgeführt werden kann.

Un sistema informatico per la gestione di patologie oncologiche croniche

L'assistenza sanitaria in Italia e nel mondo è caratterizzata da bisogni elevati in continua crescita, ad essi si contrappone l'attuale crisi delle risorse economiche determinando per il Sistema una valutazione di scelte quali la riduzione o la rimodulazione dell'offerta sanitaria pubblica. L'idea di questo lavoro, nata all'interno dell'Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, è di approcciare questo problema in ottica di miglioramento delle performance anziché riduzione dei servizi, nella convinzione che vi siano importanti margini di perfezionamento. Per questi motivi si è valutata la necessità di sviluppare un metodo e un'applicazione software per l'identificazione dei percorsi diagnostici terapeutici assistenziali (PDTA), per la raccolta di dati dalle strutture coinvolte, per l'analisi dei costi e dei risultati, mirando ad una analisi di tipo costi - efficacia e di benchmarking in ottica di presa in carico dei bisogni di salute. La tesi descrive la fase di raccolta e analisi dei requisiti comprensiva della profilazione utenti e descrizione di alcuni aspetti dinamici salienti, la fase di progettazione concettuale (schema Entity/Relationship, glossario e volumi dei dati), la fase di progettazione logica e la prototipazione dell'interfaccia utente. Riporta inoltre una valutazione dei tempi di sviluppo realizzata tramite metodologia di calcolo dei punti per caso d'uso. L'applicazione progettata è oggetto di valutazione di fattibilità presso l'IRST, che ha utilizzato alcune delle metodologie descritte nella tesi per descrivere il percorso di patologia mammaria e presentarne i primi risultati all'interno di un progetto di ricerca in collaborazione con l'Agenzia Nazionale per i Servizi Sanitari Regionali (Agenas).

Oligodendroglial exosomes in glia to neuron signaling

In the CNS, myelinating oligodendrocytes and axons form a functional unit based on intimate cell-cell interactions. In addition to axonal insulation serving to increase the conduction velocity of electrical impulses, oligodendrocytes provide trophic support to neurons essential for the long-term functional integrity of axons. The glial signals maintaining axonal functions are just at the beginning to become uncovered. Yet, their determination is highly relevant for all types of demyelinating diseases, where lack of glial support significantly contributes to pathology. rnThe present PhD thesis uncovers exosomes as a novel signaling entity in the CNS by which cargo can be transferred from oligodendrocytes to neurons. Exosomes are small membranous vesicles of endocytic origin, which are released by almost every cell type and have been implicated in intercellular communication. Oligodendrocytes secrete exosomes containing a distinct set of proteins as well as mRNA and microRNA. Intriguingly, oligodendroglial exosome release is stimulated by the neurotransmitter glutamate indicating that neuronal electrical activity controls glial exosome release. In this study, the role of exosomes in neuron-glia communication and their implications on glial support was examined. Cortical neurons internalized and accumulated oligodendroglial exosomes in the neuronal cell soma in a time-dependent manner. Moreover, uptake occurred likewise at the somatodendritic and axonal compartment of the neurons via dynamin and clathrin dependent endocytosis. Intriguingly, neuronal internalization of exosomes resulted in functional retrieval of exosomal cargo in vitro and in vivo upon stereotactic injection of Cre recombinase bearing exosomes. Functional recovery of Cre recombinase from transferred exosomes was indicated by acquired reporter recombination in the target cell. Electrophysiological analysis showed an increased firing rate in neurons exposed to oligodendroglial exosomes. Moreover, microarray analysis revealed differentially expressed genes after exosome treatment, indicating functional implications on neuronal gene expression and activity. rnTaken together, the results of this PhD thesis represent a proof of principle for exosome transmission from oligodendrocytes to neurons suggesting a new route of horizontal transfer in the CNS.rn