Os sistemas não aderentes de fixação com placas de \"granito\", em comparação com os métodos que empregam argamassas, exigem maior acuidade no projeto e uma avaliação estrutural criteriosa. As metodologias empregadas no país consideram apenas o parâmetro de resistência à tração na flexão na ruptura, não levando em conta outros parâmetros estruturais importantes. Para preencher esta lacuna, o presente trabalho propõe uma metodologia para dimensionamento de três sistemas não aderentes de fixação: fachadas ventiladas, pisos elevados e lajes estruturais. Três \"granitos\" brasileiros de alto valor comercial - Vermelho Capão Bonito (VCB), Preto São Gabriel (PSG) e Branco Desireé (BD) - foram submetidos a ensaios laboratoriais para a determinação do comportamento tensão versus deformação, da tensão de ruptura, dos deslocamentos verticais, do módulo de Young e do coeficiente de Poisson. Estes parâmetros foram empregados para o dimensionamento de fachadas ventiladas e de pisos elevados. Os modelos computacionais do programa STRAP foram calibrados com dados obtidos do monitoramento de protótipos de fachadas e de pisos elevados. Para fachadas ventiladas (placas de 60 x 100 cm) as espessuras calculadas foram de 30 mm para o VCB, de 25 mm para o PSG e de 55 mm para o BD. Para pisos elevados (placas de 60 x 60 cm) as espessuras foram de 35 mm para o VCB, de 30 mm para o PSG e de 35 mm para o BD. Os resultados sugerem que a especificação da espessura mínima de 20 mm, comumente usada em diversos projetos nacionais, não considera aspectos estruturais importantes. / Design on \"granite\" plates for non adherent covering systems, in comparison with mortar methods, needs a structural evaluation. The Brazilian design methodology considers only the breaking load bending tensile strength and do not consider another important structural characteristics. To fill this gap, this thesis presents a methodology for design on three \"granite\" plates for non adherent covering systems: ventilated façades, pedestal paving system and structural slab. Three Brazilian \"granites\" with great commercial acceptance - Capão Bonito Red (CBR), São Gabriel Black (SGB) and Desireé White (DW) were tested to obtain the strength versus strain behavior, the breaking load strength, the modulus of Young and the Poisson coefficient. To structural design this characteristics were applied in computational models, that were calibrated with tests data. The thickness calculated for ventilated façades (slab of 60 x 100 cm) is 30 mm for CBR, 25 mm for SGB and 55 mm for DW. The thickness calculated for pedestal paving system (slab of 60 x 60 cm) is 35 mm for CBR, 30 mm for SGB and 35 mm for DW. This results show that the thickness of 20mm applied for Brazilian practice design criteria do not consider important structural issues.

Este trabalho teve por finalidade analisar a importância e a qualidade percebida (desempenho) de determinados atributos, em serviços de atendimento secundário em saúde, sob a ótica de seus pacientes, visando gerar subsídios para aprovação e elaboração do planejamento estratégico e de orçamentos (investimentos) de seus serviços. O estudo foi realizado no Ambulatório Central (Amce) da Universidade de Caxias do Sul (UCS), que tem sido um ponto de apoio à população caxiense e da região na prestação de serviços nas áreas da saúde, incluindo medicina, enfermagem, psicologia, fisioterapia, nutrição e serviço social. Foi realizada uma pesquisa quantitativa com a utilização de questionários distribuídos aos pacientes em três fases do serviço do Amce (fase A - recepção e atendimento clínico, fase B realização do exame e fase C resultado do exame e revisão da consulta). As principais variáveis investigadas incluem: a facilidade de acesso ao Amce, recepção e sala de espera, estrutura física e funcional, atendimento pelo profissional da saúde, confiança na própria recuperação, aceitação de atendimento por profissional da saúde junto com alunos, tempo de espera para ser atendido e propensão para indicar o Amce a outras pessoas. Os dados foram analisados por intermédio de uma matriz importância versus desempenho, sendo que todos os construtos permaneceram no quadrante superior direito (quadrante B), para o qual é sugerido que se mantenha o bom serviço. Foram realizadas análises estatísticas adicionais, as quais mostraram que as principais variáveis em estudo apresentaram diferenças significativas entre suas médias. Com análise dos resultados foi possível detectar que há um espaço para a realização de melhorias nos serviços prestados no desempenho dos serviços ambulatoriais. / Submitted by Marcelo Teixeira (mvteixeira@ucs.br) on 2014-05-22T18:03:29Z No. of bitstreams: 1 Dissertacao Alexandre Calcagnotto.pdf: 655666 bytes, checksum: f4d497bb79461c6e26d6afc396ec2fce (MD5) / Made available in DSpace on 2014-05-22T18:03:29Z (GMT). No. of bitstreams: 1 Dissertacao Alexandre Calcagnotto.pdf: 655666 bytes, checksum: f4d497bb79461c6e26d6afc396ec2fce (MD5) / The aim of this work was to measure the importance and the quality of some specific topics (performance) offered by a secondary health care Out-patient Clinic as perceived by the patients. It had the purpose of generating data that can help to develop strategic planning to manage the clinical services. This study was carried out at the Central Out-patient Clinic (Amce) of the University of Caxias do Sul (UCS). The Amce has been playing an important role in providing health care (medicine, nursing, psychology, physiotherapy, nutrition and social service) to the population of Caxias do Sul and neighborhood areas. To implement this study a quantitative research was performed, giving a questionnaire to a sample of patients of Amce (Phase A reception and clinical assistance, phase B - carrying out of complementary exams and phase C exams results and clinical follow up). The key variables of the study included: accessibility to the Amce, reception and waiting room, building conditions, behavior of the healthcare professional, confidence on the self-recovery, acceptance to be treated by a healthcare professional and students together, quickness to handle pacient´s demands, and readiness to indicate Amce services to other people. The data was analyzed by using a matrix method correlating importance versus performance. Additional statistical analyses were carried out, which indicated significant differences between means of the main research variables. With this study it was possible to verify that the importance and performance had relevant results. Therefore, the Acme may have room to improve the rendering of health care services.

This article provides a different approach to the debated issue of the criterion of solution of what has come to be called “property not registered vs. embargo registered”. In it, it argues that all the solutions that has been raised (even the adopted as “binding precedent” in the judgment of the Seventh Civil CassationPlenary) are based on an optical error, because they look at the embargo act of a static way, as if it were an act that creates a final situation, forgetting that the embargo is a procedural act that is not an end in itself, but is inserted into the dynamic of the execution process, a process in which the final situation occurs (the awarding of the asset under seizure to the successful bidder or creditor, that is, the acquisition of a real right). Observed, however, theembargo on the dynamic of the executive procedure, as an act that prepares the forced alienation of the asset, is postulated, as a criterion of solution to the problem, that of priority registration (that is, the contained in the first paragraph of the article 2022 of the Civil Code, but with the tempering of the appreciation of good faith), once the annotation of the embargo “reserve priority” to the act of forced alienation of the asset. / El presente trabajo pretende aportar un enfoque distinto al debatido tema del criterio de solución de lo que ha venido a llamarse “propiedad no inscrita vs. embargo inscrito”. En él, se sostiene que todas las soluciones que se han planteado (incluso la adoptada como “precedente vinculante” en la Sentencia del VII Pleno Casatorio) parten de un error de óptica, pues miran el acto del embargo de manera estática, como si se tratara de un acto que crea una situación final, olvidando así que el embargo es un acto procesal que no es fin en sí mismo, sino que se inserta en la dinámica del proceso de ejecución, proceso en el cual se produce la situación final (la adjudicación del bien embargado al postor adjudicatario o al acreedoradjudicatario, esto es, la adquisición de un derecho real). Observado, en cambio, el embargo en la dinámica del procedimiento ejecutivo, como acto que prepara la enajenación forzada del bien, se postula, como criterio de solución al problema, el de la prioridad registral (esto es, la contenida en el primer párrafo del artículo 2022 del Código Civil, pero con el atemperante de la apreciación de la buena fe), en cuanto la anotación del embargo “reserva prioridad” al acto de enajenación forzada del bien.

Conselho Nacional de Desenvolvimento Científico e Tecnológico / The present paper, entitled "Technologies and their Interfaces in the Field as Strategy of Capital Control", has as objective analyzing the entrance of technologies in the agrarian space of the municipal district of Lagarto - Sergipe, as a subordination strategy and control of the work, above all, of the familiar farmer production by the capital. Starting from the Marxist critical analysis, based in the conflict capital x work-nature, it was possible to unmask the contradictions imposed by the productive restructuring of the capital in the field, in which the State is mediator of the entrance of technologies, guaranteeing the accumulation of the capital. Among the technologies that penetrated in the municipal district of Lagarto, there is a larger importance to the ones which are applied in the orange agribusiness (main agricultural product of Sergipe), and inserted in the area Center-south and in specific, the municipal district of Lagarto, starting from 2003 with the Program of Revitalization of Citricultura, by the action of the state Government. In spite of the width and of the force of the technologies found in the orange production, other present technologies in the municipal district were also analyzed: the one of production of tobacco in greenhouses under command of the company Souza Cruz; the irrigation techniques in the Irrigated Perimeter Piauí and the biotechnologies with the action of EMBRAPA. In the diffusion of those technologies, it is present the speech of the improvement of life conditions of the population of the field, through the employment generation and income, however, the study of the interfaces (two sides in which is covered the process of entrance of technologies in the field), it allowed to prove the unequal and combined movement of the capital in the limitless search for profit. The technologies in the municipal district of Lagarto, at the same time that signal the increase of productivity, they engender the spacialization in new work ways and production without, however altering in the capital cycle, the reproduction farmer's process. The redefinition of the production relations and work happens, especially, by the monopolization of the territory by the capital in the appropriation of the surplus of the unit production of farmer family, predominant in the municipal district. / O presente trabalho, intitulado As Tecnologias e suas Interfaces no Campo Como Estratégia de Controle do Capital , teve como objetivo analisar a entrada de tecnologias no espaço agrário do município de Lagarto - Sergipe, como uma estratégia de subordinação e controle do trabalho, sobretudo da unidade de produção familiar camponesa, pelo capital. A partir da análise crítica marxiana, fundamentada no conflito capital x trabalho-Natureza, foi possível desvendar as contradições impostas pela reestruturação produtiva do capital no campo, na qual o Estado é mediador da entrada de tecnologias, garantindo a acumulação do capital. Entre as tecnologias que adentram o município de Lagarto, têm maior importância as que são aplicadas no agronegócio da laranja (principal produto agrícola de Sergipe) inseridas na região Centro-Sul e em especificidade o município de Lagarto, a partir de 2003 com o Programa de Revitalização da Citricultura pela ação do Governo estadual. Apesar da amplitude e da força das tecnologias se encontrarem na produção de laranja, outras tecnologias presentes no município também foram analisadas: a da produção de fumo em estufas sob comando da empresa Souza Cruz; as técnicas de irrigação no Perímetro Irrigado Piauí e as biotecnologias com a ação da EMBRAPA. Na difusão dessas tecnologias, é presente o discurso da melhoria das condições de vida da população do campo, por meio da geração de emprego e renda, entretanto, o estudo das interfaces (dois lados em que se reveste o processo de entrada de tecnologias no campo), permitiu comprovar o movimento desigual e combinado do capital na busca ilimitada do lucro. As tecnologias no município de Lagarto, ao mesmo tempo em que sinalizam o aumento da produtividade, engendram a espacialização de novas formas de trabalho e produção sem, contudo, alterar no ciclo do capital o processo da reprodução camponesa. A redefinição das relações de produção e trabalho ocorre, especialmente, pela monopolização do território pelo capital na apropriação do excedente da unidade de produção familiar camponesa, predominante no município.

Este trabalho teve por finalidade analisar a importância e a qualidade percebida (desempenho) de determinados atributos, em serviços de atendimento secundário em saúde, sob a ótica de seus pacientes, visando gerar subsídios para aprovação e elaboração do planejamento estratégico e de orçamentos (investimentos) de seus serviços. O estudo foi realizado no Ambulatório Central (Amce) da Universidade de Caxias do Sul (UCS), que tem sido um ponto de apoio à população caxiense e da região na prestação de serviços nas áreas da saúde, incluindo medicina, enfermagem, psicologia, fisioterapia, nutrição e serviço social. Foi realizada uma pesquisa quantitativa com a utilização de questionários distribuídos aos pacientes em três fases do serviço do Amce (fase A - recepção e atendimento clínico, fase B realização do exame e fase C resultado do exame e revisão da consulta). As principais variáveis investigadas incluem: a facilidade de acesso ao Amce, recepção e sala de espera, estrutura física e funcional, atendimento pelo profissional da saúde, confiança na própria recuperação, aceitação de atendimento por profissional da saúde junto com alunos, tempo de espera para ser atendido e propensão para indicar o Amce a outras pessoas. Os dados foram analisados por intermédio de uma matriz importância versus desempenho, sendo que todos os construtos permaneceram no quadrante superior direito (quadrante B), para o qual é sugerido que se mantenha o bom serviço. Foram realizadas análises estatísticas adicionais, as quais mostraram que as principais variáveis em estudo apresentaram diferenças significativas entre suas médias. Com análise dos resultados foi possível detectar que há um espaço para a realização de melhorias nos serviços prestados no desempenho dos serviços ambulatoriais. / The aim of this work was to measure the importance and the quality of some specific topics (performance) offered by a secondary health care Out-patient Clinic as perceived by the patients. It had the purpose of generating data that can help to develop strategic planning to manage the clinical services. This study was carried out at the Central Out-patient Clinic (Amce) of the University of Caxias do Sul (UCS). The Amce has been playing an important role in providing health care (medicine, nursing, psychology, physiotherapy, nutrition and social service) to the population of Caxias do Sul and neighborhood areas. To implement this study a quantitative research was performed, giving a questionnaire to a sample of patients of Amce (Phase A reception and clinical assistance, phase B - carrying out of complementary exams and phase C exams results and clinical follow up). The key variables of the study included: accessibility to the Amce, reception and waiting room, building conditions, behavior of the healthcare professional, confidence on the self-recovery, acceptance to be treated by a healthcare professional and students together, quickness to handle pacient´s demands, and readiness to indicate Amce services to other people. The data was analyzed by using a matrix method correlating importance versus performance. Additional statistical analyses were carried out, which indicated significant differences between means of the main research variables. With this study it was possible to verify that the importance and performance had relevant results. Therefore, the Acme may have room to improve the rendering of health care services.

Ce travail étudie les composants de l’évolution territoriale et socio-économique de la région du Jari, située au nord de l’Amazonie brésilienne, en identifiant les différents processus de transformation ainsi que les structures spatiales actuelles. Longtemps à l’écart des dynamiques nationales du peuplement et du développement, celle-ci connait, au XXe siècle, un développement socioéconomique en deux étapes : Le premier, au début du siècle, à partir d’une occupation conditionnée par l’extractivisme (exploitation de ressources naturelles de la forêt, comme le caoutchouc ou la noix du Brésil), et organisée autour d’un latifundiaire sur environ 2 millions d’hectares. Le deuxième, plus important, à la fin des années 1960, à partir de l’installation, sur ce même territoire, d’une grande entreprise agro-industrielle, le "projet Jari", par l’entrepreneur et multimillionnaire Américain D.K. Ludwig, qui visait à la production de la cellulose à grande échelle, et entraînait la construction d’une première infrastructure urbaine et industrielle dans une région de forêt primaire jusque-là intacte. Désormais indissociablement liées, les transformations socioéconomiques et territoriales de la région du Jari se sont depuis produites en tant qu’influence ou interaction directe avec l’évolution du projet Jari. Celui-ci, toujours en fonction dans la région aujourd’hui, a lui-même connu des transformations significatives depuis son installation en 1967: fortement polémiquée pour la non rentabilité économique, l’impact environnemental, et le gaspillage en ressources humaines de ses activités d’agrobusiness, le projet Jari du XXIe siècle émerge aujourd’hui, sous une nouvelle gestion (Orsa), comme une entreprise viable, certifiée, et pionnière en matière de responsabilité sociale et environnementale des entreprises en contexte amazonien. Les nouvelles stratégies territoriales développées par celle-ci et basées dans un discours de responsabilité sociale et environnementale des entreprises, ont permis à la Jari non seulement de conquérir une place stable sur le marché national et international de la cellulose, mais aussi à atténuer une conflit foncier de long date dans la région du Jari, qui a durablement marqué les relations entre l’entreprise, le gouvernement et les populations locales, dont plusieurs ont été déplacées de leurs terres lors de la transformation de larges parties de forêt primaire en plantations d’eucalyptus. La création, par la "nouvelle" entreprise, d’une Fondation sociale, censée de mettre en œuvre des petits projets socio-économiques dans la région, a en outre permis d’avancer le processus d’intégration territoriale visant à inclure enfin les communautés locales et les acteurs politiques à la gestion régionale et à l’utilisation des ressources, jusque-là monopolisée par l’entreprise. Sans être achevé, ce processus forme aujourd’hui la base solide pour une transformation "durable" du Jari, d’une vieille "enclave" économique de l’Américain à une région intégrée dont le potentiel est partagé parmi ses habitants. / This thesis studies the components and phases of the socioeconomic and territorial evolution of the Jari river region, located at the north of the Brazilian Amazon, by identifying its different transformation processes and contemporary spatial structures. Longtime left aloof from national population and development dynamics, the region experiences a two-step economic and social evolution in the XX century: A first occupation, at the beginning of the century, that was conditioned by the exploitation of natural resources (rubber, Brazil nut) and led to the establishment of a "latifundium" covering about 2 million ha; and a second, more important occupation at the end of 1960ies by the an American businessman and multimillionaire D.K. Ludwig. The initiation, by the latter, of a huge agro-industrial enterprise, the "Jari project", aiming at a large-scale production of cellulose, lead to the construction of first urban infrastructures and industrial facilities in a region until then covered with intact primary forest. The socio-cultural, economical and political development of the Jari River region has since been intrinsically linked to the evolution of the Jari project. The project itself, which is still active in the region, has itself known significant change since its building up in 1967: strongly criticized during the 1970ies and 1980ies due to the lack of economic viability of its agro-industrial activities, as well as its ecological impact and its waste of human resources, the project of the XXI century, revived by a new management (Orsa), presents itself as an innovative, lucrative and certified firm, henceforth conscious of its social and environmental responsibility and thus of its pioneer role for the socioeconomic development of the Jari River region where it operates. The territorial strategies developed in this context by the company, which are embedded in a discourse of social and environmental corporate responsibility, have indeed permitted the company not only to establish itself on the national and international pulp market, but also to mitigate the long-standing land conflict between the Jari firm, the government and the local populations, who had been partly displaced from their lands by the transformation of huge parts of the rainforest into eucalyptus plantations. The creation of a firm-interne social Foundation in 2000, commissioned to carry into action small socioeconomic projects in the region, has also allowed to accelerate a process of territorial integration, aiming at including the local communities and political actors into the management of the region and its use of its natural resources, until then monopolized by the company. Not yet fully achieved, this process of territorial integration constitutes today a solid basis for a sustainable transformation of the region in the future, from the old "enclave" of the American into an integrated region whose potential is commonly shared by its habitants.

La thèse se concentre sur l’étude des impacts de la directive européenne concernant les Marchés d’Instruments Financiers (MIF) et de sa révision (MIF II et MiFIR) sur le processus de découverte des prix. Selon Schreiber et Schwartz (1986), celui-ci est défini comme l´incorporation de l´information nouvelle dans le prix des actifs et la recherche de l´équilibre par le marché. Cette directive clé a pour objectif d´augmenter la concurrence et l´efficience au niveau des marchés européens tout en assurant la protection des investisseurs, ceci via une augmentation de la transparence et une exigence de politique de meilleure exécution des ordres de la part des firmes d´investissement. Plus particulièrement, l´étude se focalise sur les actions françaises du CAC40 qui peuvent désormais être échangées en dehors du marché national réglementé, Euronext Paris. Les plateformes multilatérales de trading, les internalisateurs systématiques et les dark pools sont des alternatives qui ont été introduites par la directive. En l´absence de consolidation du marché européen dans son ensemble et en présence d’une fragmentation spatiale des ordres de bourse, le risque est que certaines places d´échanges reçoivent plus d´ordres d´achat et d´autres, plus d´ordres de vente. Certains pensent que la technologie devrait lier des marchés spatialement fragmentés. Cependant, si suffisamment de flux d´ordres est retiré du marché réglementé et transparent, ce dernier pourrait ne plus assurer la découverte des prix car les prix et les quantités d´équilibre n´auraient pas été découverts par le marché dans son ensemble. De plus, même en présence d´un marché consolidé au niveau spatial, une fragmentation temporelle peut subsister. Elle correspond à la fracturation du flux d’ordres dans le temps, rendant la rencontre des ordres d´achat et de vente plus compliquée. […] La thèse apporte tout d’abord un éclairage sur les enjeux et les implications de la directive sur l’efficience des marchés européens. Dans le premier chapitre, nous proposons un cadre d´évaluation de la directive. Une sélection des principaux travaux académiques est réalisée dans le domaine de la microstructure des marchés afin d´identifier les problématiques restant sans réponse et les enjeux pour sa révision en cours (MIF II). Ensuite, une revue de la littérature sur le processus de découverte des prix du marché est opérée par la mise en lumière des principaux travaux théoriques, méthodologiques et empiriques. Les deux principales fonctions d´un marché sont de fournir de la liquidité et de permettre la découverte des prix. Cependant, la fonction de découverte des prix a souvent été un objectif de régulation négligé par rapport aux objectifs de transparence et de concurrence. […] / The thesis focusses on the impacts of the European Markets in Financial Instruments Directive (MiFID) and its revisions (the MiFID II and the MiFIR) on the price discovery process. According to Schreiber and Schwartz (1986), the price discovery process is defined as the incorporation of new information into the prices of assets and the search for an equilibrium by the market participants. This key directive aims to increase competition and efficiency at the European level without neglecting investor protection by increasing transparency and by requiring a best execution policy for the execution of client orders from investment firms.The study specifically highlights the CAC40 stocks, which, with the implementation of the MiFID, can be exchanged outside the regulated domestic market (Euronext Paris). The directive has introduced Multilateral Trading Facilities (MTFs), Systematic Internalisers (SI) and Dark Pools as alternative trading venues.In absence of an overall consolidation of the European market and in presence of a spatial fragmentation of orders, there is a risk that some exchange places may receive more buy orders and others more sell orders. Technology should bind spatially fragmented markets; however, if enough of the order flow were removed from the regulated and transparent market, it would be unable to ensure the price discovery because the equilibrium prices and quantities would not befound by the overall market. In addition, even in the presence of a consolidated market at the spatial level, temporal fragmentation may still exist. […]The first chapter studies the challenges and the implications of the MiFID on the efficiency of the European financial markets. This research proposes a regulatory framework to assess the directive. A selection of the principal academic work in the microstructure research area has been carried out in order to identify the remaining unanswered issues and challenges for the current revision of the MIF. The second chapter proposes a literature review of the concept of price discovery by highlighting the principal theoretical, methodological and empirical academic research. The two main functions of a market are to provide liquidity and to allow price discovery. However, the price discovery function has often been a neglected regulatory objective in comparison to transparency and competition objectives. It is important to assess the impacts of fragmentation on the quality of the market after the implementation of the MiFID. The object of study is the price discovery accuracy in the post-crisis context of more high-frequency and algorithmic trading. At this level, the thesis first offers a quantification of the degree of spatial and temporal fragmentation of CAC40 shares in the post-MiFID context. This study shows an increase in fragmentation. Furthermore, the quality of the market is evaluated from a price discovery perspective through the study of an indicator developed by Ozenbas et al. (2002, 2011) called the normalised volatility ratio. The study confirms the existence of a price discovery risk at the opening of the market before and after the implementation of MiFID. The potential causes of price discovery accuracy have been studied using three types of variables that characterise each transaction. The number of ransactions and the proportion of high-frequency traders on the buy side for the first half-hour of the day are significant variables for price discovery accuracy. In the post-MiFID scenario, spatial fragmentation does not significantly affect the market quality of CAC40 shares. At this level, temporal fragmentation seems to be a greater determinant. […]

L’allogreffe de cellules hématopoïétiques est une stratégie thérapeutique importante dans les hémopathies malignes. La maladie du greffon contre l’hôte (GvH) en est une complication majeure menaçant le pronostic vital. Elle est due à la reconnaissance des antigènes du receveur par les lymphocytes T du donneur et à l’activation de ceux-ci, à l’origine de dommages tissulaires. L’altération de la barrière intestinale joue un rôle critique dans la GvH. La famille des protéines de choc thermique (HSP)90 comporte cinq membres dont trois cytosoliques dénommés HSP90, et un localisé dans le réticulum endoplasmique (RE), Gp96, qui peut être sécrété en cas de stress. Nous montrons dans nos travaux de thèse que la 17AAG, un inhibiteur des HSP90, réduit la mortalité liée à la GvH dans un modèle murin. Cet effet est associé à une augmentation de la réponse au stress du RE dans les cellules épithéliales intestinales comme en atteste l’augmentation de l’épissage du facteur de transcription XBP-1, corrélée à une diminution du dommage tissulaire intestinal. Ces résultats permettent d’envisager une place pour la 17AAG ou d’autres inhibiteurs de HSP90 dans la prévention de la GvH chez l’homme. D’autre part, nous montrons que Gp96 est sécrétée dans le sérum de patients développant une GvH aiguë sévère avec atteinte intestinale. Nous suggérons de valider la pertinence de Gp96 comme biomarqueur de GvH intestinale dans une étude de plus grande ampleur. Enfin, nous trouvons que Gp96 s’associe avec le composant 3 du complément, une protéine impliquée dans l’immunité innée et adaptative, et inhibe certaines de ses fonctions. Les conséquences fonctionnelles de cette association sont discutées. / Allogeneic hematopoietic cell transplantation is a treatment for certain disorders including hematologic malignancies. Graft-versus-host-disease (GvHD) is a major, life-threatening complication. It is due to the recognition of recipient antigens by donor T cells, which activate and damage tissues. Intestinal barrier alteration plays a critical role in GvHD. Heat shock proteins (HSP)90 include five members, three cytosolic members named HSP90, and one member localized in endoplasmic reticulum (ER) called Gp96 and able to gain extracellular level in case of stress. We show in our thesis that 17AAG, a HSP90 inhibitor, reduces GvHD mortality in a mouse model. This effect is associated with an increase in ER stress pathway in intestinal epithelial cells as figured by transcription factor XBP-1 splicing, correlated to a decrease in intestinal tissue damage. These results suggest that 17AAG could be considered in GvHD prevention in human. Moreover, we show that Gp96 is secreted in serum of patients developing an acute GvHD with intestinal involvement. We propose to validate the relevance of Gp96 as an intestinal GvHD biomarker is a larger study. Finally, we find that Gp96 associate with complement component 3, a protein involved in innate and adaptive immunity, and inhibit some of its functions. Functional consequences of this association are discussed.

On admet communément que les tumeurs proviennent de cellules échappant aux contrôles homéostatiques qui sous-tendent les structures histologiques saines et que le phénotype d’une cellule n’est pas le résultat de processus génétiques et biochimiques déterministes mais la conséquence stochastique de réseaux de régulation intra- et intercellulaires. Ce doctorat vise à étudier quantitativement l’homéostasie phénotypique de populations cellulaires et à présenter une approche à la question fondamentale, mais jusqu’alors jamais étudiée, concernant l’autonomie versus le contrôle collectif du devenir des cellules. Nous avons étudié sur le long terme, par cytométrie de flux et dans des conditions 2D puis 3D, le niveau d’expression de CD24 et CD44 de deux lignées cellulaires de cancer du sein (SUM149-PT et SUM159-PT). Trois phénotypes ont été isolés (CD24-/CD44+, CD24+/CD44+, CD24-/CD44-), ce dernier n’avait pour le moment pas été documenté dans la littérature. Le comportement phénotypique des sous-populations CD44-low et CD44-high a été caractérisé en évaluant leur proportion et en analysant leur spectre de fluorescence. Ainsi nous avons observé des comportements périodiques d’apparition et de disparition de pool de cellules caractéristiques des lignées et une re-diversification des phénotypes pour chacune des sous-population. Seule la population issue de CD24-/CD44- re-diversifiée présente le même équilibre que la population initiale non triée. En 3D, le processus de re-diversification a été observé dans les tumorsphères issues de CD24-/CD44+ et CD24+/CD44+. Les cellules CD24-/CD44- n’ont pas ce potentiel mais survivent néanmoins à l’anoïkis. Ces comportements laissent penser qu’il existe une coordination intercellulaire régulant l’équilibre des proportions phénotypiques. Pour découvrir les règles sociales régissant l’organisation spatiale inter-phénotypique, nous avons mis en place un rapporteur des variations du niveau d’expression endogène des marqueurs d’intérêt et élaboré un modèle théorique d’interactions cellulaires. Ce travail a conforté notre hypothèse selon laquelle il s’établit des règles sociales inter-cellulaires déterminant l’expression phénotypique à l’échelle uni- et pluricellulaire. / It is commonly accepted that tumors arise from cells that escape the homeostatic controls which underlie the healthy histological structure and that cell phenotype is not the result of deterministic biochemical and genetic processes, but rather the stochastic and dynamic outcome of multiple intra- and intercellular regulation networks. This PhD aims to quantitatively study the phenotypic homeostasis of the cell populations and to present an approach to the fundamental question, never heretofore studied, regarding the autonomy versus collective control of cell fate. We studied in the long run, using flow cytometry and in 2D and 3D conditions, the level of expression of CD24 and CD44 of two breast cancer cell lines (SUM149-PT and SUM159-PT). Three phenotypes were isolated (CD24-/CD44+, CD24+/CD44+, CD24-/CD44-), the latter had not previously been documented in the literature. The phenotypic behavior of CD44-low and CD44-high subpopulations has been characterized by assessing their proportion and analyzing the fluorescence map. Thereby, we observed both a periodic behavior of appearance and disappearance of pool of cells characteristics of each cell lines and a phenotypic re-diversification for each subpopulation. Only the resulting population derived from CD24-/CD44- provided the same balance as the original unsorted population. 3D re-diversification process was observed in tumorspheres from CD24-/CD44+ and CD24+/CD44+. The cells CD24-/CD44did not have that potential but nonetheless outlived anoikis. These behaviors suggest that there is an inter-cell coordination regulating the balance of phenotypic proportions. To discover the social rules regulating inter-phenotypic spatial organization, we have set up a reporter of the endogenous variations of CD24 and CD44 and developed a theoretical model of cell interactions. This work has confirmed our hypothesis that inter-cellular social rules are determining the phenotypic expression at both the uni- and multicellular scales.

Indiana University-Purdue University Indianapolis (IUPUI) / Graft-versus-host disease (GVHD) remains the major complication after allogeneic hematopoietic stem cell transplantation (HSCT), resulting from immunological attack on target organs such as gastrointestinal (GI) tract, liver and skin from donor allogeneic T cells. The most common treatment for GVHD is immunosuppressive drugs such as corticosteroids, which may result in many side effects including the loss of the beneficial graft-versus-leukemia (GVL) effect and increased infection rates. However, GVHD-specific drugs have yet to be implemented. Here we show that by targeting on a novel pathogenic CD4+ T cell subpopulation that our lab previously found in patients with GI GVHD, we can develop new avenues to treat GVHD. This novel population is characterized as CD146+CCR5+ T cells, co-expressing IL-17A and IFN-γ. We found that the inducible T-cell costimulator (ICOS), which has been reported to be important for human Th17 differentiation in vitro, is critical for the development of this nonconventional T Helper 1 (Th1*)-polarized CD146+CCR5+ conventional T cells (Tconvs) population. Furthermore, we found that ICOS can induce the generation of Th1*-polarized CD146+CCR5+ regulatory T cells (Tregs) population, lowering the frequencies of phenotypic markers of functional Tregs. Our data also showed that inhibiting the major transcriptional factor of Th17, RAR-related orphan receptor gamma t (RORγt), could prevent the development of CD146+CCR5+ Tconvs in vitro. Our results demonstrate how pathogenic CD146+CCR5+ T cells are induced through ICOS or RORγt, suggesting new targets for GVHD treatment. We anticipate our assay to be a starting point for the development of novel GVHD-specific drugs. For example, the treatments that focus on inhibiting RORγ would have fewer side effects than general immunosuppressive drugs that GVHD patients use today and inhibit GVHD while sparing the GVL effect. Furthermore, we expect the CD146+CCR5+ Tconvs and/or Tregs can be used as GVHD biomarkers. These biomarkers may guide preemptive treatments such as RORγt inhibitor.