Fenomenologia del concetto di élite e network analysis come metodo di information retrieval

According to Sartori, “Language is the sine-qua-non instrument of knowing”. Nevertheless, social sciences often adopt terms lacking clear and precise definitions. Adopting the method proposed in Sartori (1984), the present Master’s thesis consists of an explicatory study aiming to clarify the concept of “elites”. First, the essential works concerning elites are identified by analyzing a large citation network. Then, the thesis proposes a definition of elite able to encompass those of previous works. The new definition attempts to include the concept of elites within the broader framework of the resource-dependence theories (Pfeffer e Salancik, 1978) and purposive theories (Coleman, 2005)

Tecniche di biologia molecolare nello studio dei sistemi chemosensoriali

TECNICHE DI BIOLOGIA MOLECOLARE NELLO STUDIO DEI SISTEMI CHEMOSENSORIALI RIASSUNTO La caratterizzazione ultrastrutturale a partire dagli anni sessanta e in seguito recentemente quella immunoistologica ha permesso di identificare cellule specializzate che possiedono diverse analogie morfologiche e molecolari con le cellule gustative ma non sono organizzate in calici gustativi. Queste cellule chemosensoriali solitarie (CCS) sono molto diffuse oltre che nella cavità orale anche nelle vie aeree e nell’apparato digerente. L’identificazione di queste cellule singole o disposte in cluster ha portato a concettualizzare l’esistenza di un sistema chemosensoriale diffuso (DCS) di cui le cellule gustative organizzate nei taste buds nella cavità orale sono solo la parte più evidente. Le funzioni del DCS sembrano coinvolgere la secrezione, la risposta immunitaria innata, l’assorbimento ma altri ruoli sono in parte ancora speculativi, e inoltre non tutti i tipi di cellule sono stati caratterizzati, lasciando aperte molte questioni che questo studio ha in parte analizzato utilizzando tecniche di biologia molecolare. In particolare sono state indagate tramite RT-PCR e western blot le differenti capacità chemorecettoriali degli organi di origine endodermica ricercando l’espressione genica dei recettori implicati nella percezione del gusto. Inoltre sono state coadiuvate ricerche finalizzate a meglio caratterizzare le cellule recettoriali all’interno dei taste buds, che risultano possedere un ventaglio di pathway molecolari molto ampio, comprendente elementi tipici delle vie aeree e dell’apparato gastrointestinale. È stato analizzato se l’espressione degli stessi geni sia modificabile a livello intestinale da diversi tipi di diete. I dati raccolti indicano una differente localizzazione dei recettori del dolce e dell’amaro lungo le vie aeree che hanno un ruolo di modulazione del movimento ciliare e della secrezione. Mentre la presenza dei recettori del dolce non va in maggior profondità della trachea, alcuni recettori dell’amaro risultano espressi anche nei polmoni. Invece lungo l’apparato gastrointestinale i recettori dell’amaro sembrano essere espressi in maniera selettiva e differenziata. L’intestino risulta inoltre sensibile a diete prolungate sovra esprimendo o sotto regolando l’espressione di proteine coinvolte nell’assorbimento e nella secrezione. In particolare è stato approfondito il sensing intestinale verso gli acidi grassi mediante quantificazione tramite PCR real time. I risultati indicano che diete iperlipidiche croniche abbattono sia l’espressione di CD36, una proteina con alta affinità per gli acidi grassi presente nella membrana apicale degli enterociti, sia l’espressione dei meccanismi di segnalazione di “saziazione” indotta dai grassi. Si tratta di un segnale, che diversamente da altri ormoni intestinali ha la capacità di prolungare la latenza tra i pasti, viene generato principalmente dalla produzione dell’ormone lipidico oleoil-etanolamide (OEA). Questo ulteriore studio oltre a evidenziare un nuovo aspetto negativo delle diete iperlipidiche, trova l’esistenza di una correlazione tra i diversi geni indagati che suggerisce che CD36 funga come sensore intestinale degli acidi grassi e abbia ruoli regolatori sui meccanismi di produzione di OEAMOLECULAR BIOLOGY TECHNIQUES IN CHEMOSENSORY SYSTEMS STUDY ABSTRACT Since 1960s, the ultrastructural and more recently the immunohystological characterization have identified specialized cells with several morphological and molecular similarities with the taste cells, without being organized into taste buds. These solitary chemosensory cells (CCS) are widespread not only in the oral cavity but even in the respiratory and gastrointestinal tract. The identification of these cell, that can be individually arranged or clustered, has led to conceptualize the existence of a diffuse chemosensory system (DCS) where the taste receptor cells organized within taste buds in the oral cavity are only the most obvious. The functions appear to involve the secretion and absorption of the DCS, and the innate immune response, but other roles are still partly speculative as not all cell types have been characterized, leaving opened many questions that this study has partly analyzed using molecular biology techniques. In particular, the different chemoreceptorial capacities of endodermal origin organs were assessed by RT-PCR and western blot examining the gene expression of those receptors that are involved in taste perception. We also performed experiments to better characterize the receptor cells within taste buds, showing that they have a very wide range of molecular pathways including typical features of the respiratory and gastrointestinal tract. It was analyzed whether the expression of these genes is modified by different types of diets at intestinal level. The data indicate a different localization of the bitter and sweet receptors along the respiratory pathway that have a role in the modulation of ciliary movement and secretion. Whereas the sweet receptors cannot be found deeper than trachea, some of the bitter receptors are also expressed in the lungs. On the other hand, along the gastrointestinal tract the bitter receptors appear to be expressed in a selective and differentiated manner. The intestine is also sensitive to prolonged diets, overexpressing or downregulating the expression of proteins involved in absorption and secretion. In particular, we studied the intestinal fatty acid sensing by real time PCR utilize. The results indicate that chronic high-fat diets cut down both the expression of CD36, a protein with high affinity for fatty acids present in the apical membrane of enterocytes, and the expression of the signaling mechanisms of satiation induced by fat. This signal, which unlike other intestinal hormones has the ability to prolong the latency between meals, is generated mainly by the production of the lipid hormone oleoylethanolamide (OEA). This study further highlights a new additional disadvantage of high-fat diets and the existence of a correlation between different genes, suggesting that CD36 act as a fatty acids sensor and have regulator roles upon the intestinal mechanisms of OEA production

Loss of the candidate tumor suppressor ZEB1 (TCF8, ZFHX1A) in Sézary syndrome

Cutaneous T-cell lymphoma is a group of incurable extranodal non-Hodgkin lymphomas that develop from the skin-homing CD4+ T cell. Mycosis fungoides and Sézary syndrome are the most common histological subtypes. Although next-generation sequencing data provided significant advances in the comprehension of the genetic basis of this lymphoma, there is not uniform consensus on the identity and prevalence of putative driver genes for this heterogeneous group of tumors. Additional studies may increase the knowledge about the complex genetic etiology characterizing this lymphoma. We used SNP6 arrays and GISTIC algorithm to prioritize a list of focal somatic copy-number alterations in a dataset of multiple sequential samples from 21 Sézary syndrome patients. Our results confirmed a prevalence of significant focal deletions over amplifications: single well-known tumor suppressors, such as TP53, PTEN, and RB1, are targeted by these aberrations. In our cohort, ZEB1 (TCF8, ZFHX1A) spans a deletion having the highest level of significance. In a larger group of 43 patients, we found that ZEB1 is affected by deletions and somatic inactivating mutations in 46.5% of cases; also, we found potentially relevant ZEB1 germline variants. The survival analysis shows a worse clinical course for patients with ZEB1 biallelic inactivation. Multiple abnormal expression signatures were found associated with ZEB1 depletion in Sézary patients we verified that ZEB1 exerts a role in oxidative response of Sézary cells. Our data confirm the importance of deletions in the pathogenesis of cutaneous T-cell lymphoma. The characterization of ZEB1 abnormalities in Sézary syndrome fulfils the criteria of a canonical tumor suppressor gene. Although additional confirmations are needed, our findings suggest, for the first time, that ZEB1 germline variants might contribute to the risk of developing this disease. Also, we provide evidence that ZEB1 activity in Sézary cells, influencing the reactive oxygen species production, affects cell viability and apoptosis

Hospital de Clinicas da UFPR : desafios para implantação de um plano estratégico

Orientador : Clverson Renan da CunhaMonografia (especialização) - Universidade Federal do Paraná, Setor de Ciências Sociais Aplicadas, Curso de Especialização MBA em Gestão EstratégicaInclui referênciasResumo : O Hospital de Clínicas da Universidade Federal é um dos maiores hospitais públicos do país. Uma instituição federal altamente influenciada pelo cenário externo, principalmente pelas ações, projetos e políticas do governo federal. Uma administração estratégica, para uma organização complexa como o Hospital, é essencial para uma gestão eficiente num ambiente altamente instável e turbulento. Permite que o gestor se centre no controle da direção da instituição, buscando a melhoria do seu desempenho criando um ambiente propício para a pesquisa e inovação e garantindo a sustentabilidade do Hospital. Porém, essa mesma condição, frustrou a consecução do plano estratégico 2012-2015, justamente no momento da sua implantação. Analisando o processo de implantação do Plano elaborado para o período de 2012-2015, deverá buscar formas de superar as dificuldades apresentadas e meios para concluir a implantação do próximo plano, de forma que possibilite uma avaliação e análise dos resultados atingidos

T Cell Leukemia/Lymphoma 1A is essential for mouse epidermal keratinocytes proliferation promoted by insulin-like growth factor 1

T Cell Leukemia/Lymphoma 1A is expressed during B-cell differentiation and, when overexpressed, acts as an oncogene in mouse (Tcl1a) and human (TCL1A) B-cell chronic lymphocytic leukemia (B-CLL) and T-cell prolymphocytic leukemia (T-PLL). Furthermore, in the murine system Tcl1a is expressed in the ovary, testis and in pre-implantation embryos, where it plays an important role in blastomere proliferation and in embryonic stem cell (ESC) proliferation and self-renewal. We have also observed that Tcl1-/-adult mice exhibit alopecia and deep ulcerations. This finding has led us to investigate the role of TCL1 in mouse skin and hair follicles. We have found that TCL1 is expressed in the proliferative structure (i.e.The secondary hair germ) and in the stem cell niche (i.e.The bulge) of the hair follicle during regeneration phase and it is constitutively expressed in the basal layer of epidermis where it is required for the correct proliferative-differentiation program of the keratinocytes (KCs). Taking advantage of the murine models we have generated, including the Tcl1-/-and the K14-TCL1 transgenic mouse, we have analysed the function of TCL1 in mouse KCs and the molecular pathways involved. We provide evidence that in the epidermal compartment TCL1 has a role in the regulation of KC proliferation, differentiation, and apoptosis. In particular, the colony-forming efficiency (CFE) and the insulin-like growth factor 1 (IGF1)-induced proliferation are dramatically impaired, while apoptosis is increased, in KCs from Tcl1-/-mice when compared to WT. Moreover, the expression of differentiation markers such as cytokeratin 6 (KRT6), filaggrin (FLG) and involucrin (IVL) are profoundly altered in mutant mice (Tcl1-/-). Importantly, by over-expressing TCL1A in basal KCs of the K14-TCL1 transgenic mouse model, we observed a significant rescue of cell proliferation, differentiation and apoptosis of the mutant phenotype. Finally, we found TCL1 to act, at least in part, via increasing phospho-ERK1/2 and decreasing phospho-P38 MAPK. Hence, our data demonstrate that regulated levels of Tcl1a are necessary for the correct proliferation and differentiation of the interfollicular KC

Gênese e metagênese: o artista e o geneticista em busca da fonte

Pretende-se investigar a complexa, mas profícua, rede de interconexões que subjazem à criação do balé La rosa del deserto idealizado pela artista plástica italiana Silva Cavalli Felci. No mesmo espaço criativo e, no meio de uma densa teia de estímulos procedentes de vários signos, convivem artes plásticas,fotografia, dança, desenho, música,coreografias, figurinos, cenografia,concretizando um corpus privilegiado para o estudo do processo criativo da obra de arte. A artífice dessa complexa rede perceptiva e criativa é a artista plástica Silva Cavalli Felcique, além de conceder o material que constitui o prototexto em apreço, torna-seco-pesquisadora da sua criação, em um processo metagenético

The relationship between Unicamp and social segments under the perspective of extension courses

Orientador: Milena Pavan SerafimDissertação (mestrado) - Universidade Estadual de Campinas, Instituto de GeociênciasResumo: O objetivo da pesquisa é investigar como se dá a relação entre a Universidade Estadual de Campinas (Unicamp) e os diferentes setores da sociedade através dos cursos de extensão. Três perguntas nos motivam: 1) a quem ¿ "quais atores" ¿ e com quais temas ¿ "quais problemas sociais e interesses" ¿ este tipo de extensão está comprometida? 2) Essa modalidade extensionista se aproxima de quais perspectivas conceituais e históricas da extensão? 3) Como esta modalidade se insere na totalidade extensionista da Unicamp? Como referencial teórico, utilizou-se diversas bibliográficas acadêmicas sobre a extensão universitária a fim de traçar sua trajetória histórica no Brasil, identificando nesta a própria origem e surgimento dos cursos de extensão ¿ também chamados de práticas de educação continuada. Além do mais, identificou-se uma série de concepções, práticas e conceitos extensionistas: extensão enquanto prestação de serviços e relação entre universidade, empresa e mercado; enquanto mecanismo de divulgação e difusão do conhecimento científico; enquanto práticas assistenciais e voluntaristas; a extensão enquanto relação dialógica entre a universidade e a classe trabalhadora; e os conceitos referentes à extensão enquanto função articuladora do ensino e da pesquisa com buscas a estabelecer relação dialógica e horizontal com segmentos sociais com vistas ao cumprimento do compromisso social da universidade. Com efeito, sob a posse deste referencial teórico e histórico, empreendeu-se uma descrição e análise dos cursos de extensão da Unicamp, bem como da própria atuação da Escola de Extensão (EXTECAMP) enquanto organização que gerencia e divulga esta modalidade. Metodologicamente, adotou-se diversos procedimentos. O primeiro deles foi uma ampla revisão nos documentos e normas da Unicamp e da EXTECAMP ¿ como estatutos, deliberações, relatórios, site institucional ¿ para descrevermos e analisarmos o objeto de estudo e traçarmos suas características e, assim, responder as perguntas de pesquisa. Empreendeuse, ainda através de revisão documental, uma análise quantitativa da evolução dos cursos de extensão ao longo do tempo, no que se refere ao número de cursos, arrecadação, perfil do aluno e institutos e faculdades que mais se destacam na oferta e arrecadação. Além disto, aplicou-se um questionário aos docentes que ministram cursos de extensão e aos funcionários que trabalham com o tema para identificar suas percepções extensionistas e, assim, melhor compreender os motivos pelos quais ofertam cursos. Como resultados, basicamente, descobriu-se que os cursos se alinham as concepções extensionistas relativos à divulgação científica e transferência de conhecimentos técnicos e à prestação de serviços de formação utilitária ao mercado e a profissões liberais. Assim, apesar de cada área do conhecimento possuir especificidades no que se refere aos segmentos e interesses sociais pelos com os quais se conectam, percebeu-se inclinação, por parte dos docentes e da própria EXTECAMP, a privilegiar interesses formativos ao mercado e ao setor produtivo. Além do mais, constatou-se que esta modalidade: não possuí vínculos formais com as outras duas funções universitárias; possuem grau de ofertismo elevado; são legitimados academicamente pois captam recursos ¿ que são usados pelos institutos como "quebra-galho" orçamentário, permitem o acesso ao ensino de indivíduos excluídos da graduação convencional e ajudam a cumprir "a missão social da Unicamp"Abstract: The objective of the research is to investigate the relationship between the State University of Campinas (Unicamp) and the different sectors of society through extension courses. Three questions motivate us: 1) whom - "which actors" - and with what themes - "what social problems and interests" - is this type of extension compromised? 2) This extensionist approach approaches which conceptual and historical perspectives of extension? 3) How does this modality fit into the total extension of Unicamp? As a theoretical reference, several academic bibliographies on university extension were used to trace their historical trajectory in Brazil, identifying in this the very origin and appearance of extension courses ¿ also called continuing education practices. Moreover, a series of extensionist conceptions, practices and concepts were identified: extension as service rendering and relationship between university, company and market; as a mechanism for disseminating and disseminating scientific knowledge; as caring and voluntary practices; extension as a dialogical relationship between the university and the working class; And the concepts related to extension as an articulating function of teaching and research with a search to establish a dialogical and horizontal relationship with social segments in order to fulfill the social commitment of the university. After this, a description and analysis of the extension courses of Unicamp was carried out, as well as the own performance of the School of Extension (EXTECAMP) as an organization that manages and disseminates this modality. Methodologically, several procedures were adopted. The first of these was a broad revision in the documents and standards of Unicamp and EXTECAMP ¿ such as statutes, deliberations, reports, institutional site ¿ to describe and analyze the object of study and to trace its characteristics and thus answer the research questions. A quantitative analysis of the evolution of extension courses over time was also carried out through a documentary review. In addition, a questionnaire was applied to teachers who teach extension courses and to staff who work with the subject to identify their extensionist perceptions, and thus, to better understand the reasons for which they offer courses. As a result, it was basically found that the courses aligned with extensionist conceptions related to scientific dissemination and transfer of technical knowledge and to the provision of utilitarian training services to the market and to the liberal professions. Thus, although each area of knowledge has specificities in terms of the segments and social interests by which they connect, there was an inclination, on the part of teachers and of EXTECAMP itself, to privilege formative interests to the market and to the productive sector. Moreover, it was found that this modality: has no formal ties with the other two university functions; Have a high degree of offer; Are legitimized academically because they capture resources - which are used by institutes as a budget "break", allow access to the education of individuals excluded from conventional graduation and help fulfill "the social mission of Unicamp"MestradoPolitica Cientifica e TecnologicaMestre em Política Científica e TecnológicaCAPE

Establishing virtual bioequivalence and clinically relevant specifications using in vitro biorelevant dissolution testing and physiologically-based population pharmacokinetic modeling. Case example:Naproxen

Background Physiologically-based population pharmacokinetic modeling (popPBPK) coupled with in vitro biopharmaceutics tools such as biorelevant dissolution testing can serve as a powerful tool to establish virtual bioequivalence and set clinically relevant specifications. One of several applications of popPBPK modeling is in the emerging field of virtual bioequivalence (VBE), where it can be used to streamline drug development by implementing model-informed formulation design and to inform regulatory decision-making e.g., with respect to evaluating the possibility of extending BCS-based biowaivers beyond BCS Class I and III compounds in certain cases. Methods In this study, Naproxen, a BCS class II weak acid was chosen as the model compound. In vitro biorelevant solubility and dissolution experiments were performed and the resulting data were used as an input to the PBPK model, following a stepwise workflow for the confirmation of the biopharmaceutical parameters. The naproxen PBPK model was developed by implementing a middle-out approach and verified against clinical data obtained from the literature. Once confidence in the performance of the model was achieved, several in vivo dissolution scenarios, based on model-based analysis of the in vitro data, were used to simulate clinical trials in healthy adults. Inter-occasion variability (IOV) was also added to critical physiological parameters and mechanistically propagated through the simulations. The various trials were simulated on a “worst/best case” dissolution scenario and average bioequivalence was assessed according to Cmax, AUC and Tmax. Results VBE results demonstrated that naproxen products with in vitro dissolution reaching 85% dissolved within 90 min would lie comfortably within the bioequivalence limits for Cmax and AUC. Based on the establishment of VBE, a dissolution “safe space” was designed and a clinically relevant specification for naproxen products was proposed. The interplay between formulation-related and drug-specific PK parameters (e.g., t1/2) to predict the in vivo performance was also investigated. Conclusion Over a wide range of values, the in vitro dissolution rate is not critical for the clinical performance of naproxen products and therefore naproxen could be eligible for BCS-based biowaivers based on in vitro dissolution under intestinal conditions. This approach may also be applicable to other poorly soluble acidic compounds with long half-lives, providing an opportunity to streamline drug development and regulatory decision-making without putting the patient at a risk

Establishing virtual bioequivalence and clinically relevant specifications using in vitro biorelevant dissolution testing and physiologically-based population pharmacokinetic modeling. Case example:Naproxen

Background: Physiologically-based population pharmacokinetic modeling (popPBPK) coupled with in vitro biopharmaceutics tools such as biorelevant dissolution testing can serve as a powerful tool to establish virtual bioequivalence and set clinically relevant specifications. One of several applications of popPBPK modeling is in the emerging field of virtual bioequivalence (VBE), where it can be used to streamline drug development by implementing model-informed formulation design and to inform regulatory decision-making e.g., with respect to evaluating the possibility of extending BCS-based biowaivers beyond BCS Class I and III compounds in certain cases. Methods: In this study, Naproxen, a BCS class II weak acid was chosen as the model compound. In vitro biorelevant solubility and dissolution experiments were performed and the resulting data were used as an input to the PBPK model, following a stepwise workflow for the confirmation of the biopharmaceutical parameters. The naproxen PBPK model was developed by implementing a middle-out approach and verified against clinical data obtained from the literature. Once confidence in the performance of the model was achieved, several in vivo dissolution scenarios, based on model-based analysis of the in vitro data, were used to simulate clinical trials in healthy adults. Inter-occasion variability (IOV) was also added to critical physiological parameters and mechanistically propagated through the simulations. The various trials were simulated on a “worst/best case” dissolution scenario and average bioequivalence was assessed according to Cmax, AUC and tmax. Results: VBE results demonstrated that naproxen products with in vitro dissolution reaching 85% dissolved within 90 minutes would lie comfortably within the bioequivalence limits for Cmax and AUC. Based on the establishment of VBE, a dissolution “safe space” was designed and a clinically relevant specification for naproxen products was proposed. The interplay between formulation-related and drug-specific PK parameters (e.g., t1/2) to predict the in vivo performance was also investigated. Conclusion: Over a wide range of values, the in vitro dissolution rate is not critical for the clinical performance of naproxen products and therefore naproxen could be eligible for BCS-based biowaivers based on in vitro dissolution under intestinal conditions. This approach may also be applicable to other poorly soluble acidic compounds with long half-lives, providing an opportunity to streamline drug development and regulatory decision-making without putting the patient at a risk

Purification, characterization and molecular cloning of the major chitinase from Tenebrio molitor larval midgut

Insect chitinases are involved in degradation of chitin from the exoskeleton cuticle or from midgut peritrophic membrane during molts. cDNAs coding for insect cuticular and gut chitinases were cloned, but only chitinases from moulting fluid were purified and characterized. In this study the major digestive chitinase from T. molitor midgut (TmChi) was purified to homogeneity, characterized and sequenced after cDNA cloning. TmChi is secreted by midgut epithelial cells, has a molecular weight of 44 kDa and is unstable in the presence of midgut proteinases. TmChi shows strong substrate inhibition when acting on umbelliferyl-derivatives of chitobio- and chitotriosaccharides, but has normal Michaelis kinetics with the N-acetylglucosamine derivative as substrate. TmChi has very low activity against colloidal chitin, but effectively converts oligosaccharides to shorter fragments. The best substrate for TmChi is chitopentaose, with highest kcat/KM value. Sequence analysis and chemical modification experiments showed that the TmChi active site contains carboxylic groups and a tryptophane, which are known to be important for catalysis in family 18 chitinases. Modification with p-hidroximercuribenzoate of a cysteine residue, which is exposed after substrate binding, leads to complete inactivation of the enzyme. TmChi mRNA encodes a signal peptide plus a protein with 37 kDa and high similarity with other insect chitinases from family 18. Surprisingly, this gene does not encode the C-terminal Ser-Thr-rich connector and chitin-binding domain normally present in chitinases. The special features of TmChi probably result from its adaptation to digest chitin-rich food without damaging the peritrophic membrane. © 2006 Elsevier Ltd. All rights reserved