Introdução: A disfunção neurogênica do trato urinário inferior (DNTUI) é comum, tendo como causa diversas doenças neurológicas e se apresentando de maneira diversa e heterogênea. O tratamento da DNTUI pode incluir medicamentos, utilização de toxina botulínica intravesical e tratamento cirúrgico. A cirurgia de ampliação vesical é uma alternativa consagrada, usada principalmente no caso de bexigas fibrosadas ou em pacientes refratários ao tratamento conservador ou minimamente invasivo. Os mecanismos que levam à progressão da fibrose vesical e à refratariedade aos tratamentos conservadores nessa população de pacientes não são bem conhecidos. Objetivos: Avaliar a expressão gênica dos fatores relacionados à hipóxia e fibrose nos pacientes com DNTUI e correlacionar o padrão de expressão gênica com as características urodinâmicas dos pacientes estudados. Métodos: Foram avaliados prospectivamente 17 pacientes com DNTUI que foram submetidos à cirurgia de ampliação vesical pelo Departamento de Urologia do HCFMUSP. Os pacientes foram avaliados de acordo com os sintomas clínicos, exames laboratoriais e avaliação urodinâmica completa. Durante a cirurgia foi obtido um fragmento vesical interessando todas as camadas da bexiga para análise de expressão gênica com utilização de qRT-PCR de MMP-1, TIMP-1, HIF1alfa, HIF2alfa, HIF3alfa, iNOS, eNOS, VEGF e CTGF. Amostras vesicais de 9 doadores cadavéricos foram utilizados como controles. Resultados: A média dos pacientes estudados foi de 37,5 anos, com complacência vesical variando entre 3,8 e 50 ml/cmH20, com média de 11,16ml/cmH20. Os pacientes apresentaram superexpressão estatisticamente significativa de TIMP-1 e HIF3alfa e subexpressão estatisticamente significativa de MMP-1. A expressão gênica de HIF1alfa e HIF2alfa também apresentou uma tendência à superexpressão, apesar de não ser estatisticamente significativa (p = 0,14 e p = 0,11). Os outros genes avaliados foram expressos de forma heterogênea. Conclusão: Em pacientes com DNTUI associados a fibrose vesical e/ou refratariedade ao tratamento conservador, que foram submetidos à cirurgia de ampliação vesical, os fatores relacionados ao aumento da deposição da matriz extracelular e à hipóxia parecem ser relevantes na progressão da disfunção vesical. Esse achados são compatíveis com estudos em modelos animais de obstrução infravesical e hipóxia e reforçam a necessidade de estudos complementares para o melhor entendimento da fisiopatologia da progressão da disfunção vesical na DNTUI / Introduction: The neurogenic lower urinary tract dysfunction (NLUTD) is a common complication of a variety of neurological diseases and can present itself in a diverse and heterogeneous way. NLUTD can be treated with anticholinergic or alfa3 agonists drugs, intravesical botulinum toxin and/or surgical treatment. The bladder augmentation surgery is a classic alternative, used mainly in cases of fibrotic bladders or in patients with refractory symptoms with conservative treatment. The mechanisms that lead to the progression of bladder fibrosis and to conservative treatments failure are not well known. Objectives: To evaluate the expression of fibrosis and hypoxia related genes in patients with NLTUD and correlate the pattern of gene expression with urodynamic data of the population studied. Methods: We analyzed bladder specimens of 17 patients with NLUTD undergoing bladder augmentation surgery due to bladder fibrosis or conservative treatment failure. Urodynamic tests were performed in all patients. A bladder fragment was obtained during surgery for relative gene epression with quantitative real-time polymerase chain reaction (qRT-PCR) of MMP-1, TIMP-1, HIF1alfa, HIF2alfa, HIF3alfa, iNOS, eNOS, VEGF e CTGF. 9 cadaveric organ donors composed the control group. Results: The mean age of the patients was 37.5 years. Bladder compliance ranged form 3,8 to 50 ml/cmH20, with a mean of 11,16 ml/cmH20. Patients undergoing bladder augmentation surgery presented a statistically significant overexpression of TIMP-1 and HIF3alfa. MMP-1 was statistically significant underexpressed. The gene expression of HIF1alfa and HIF2alfa also showed a tendency to overexpression, although it was not statistically significant (p = 0,14 and p = 0,11). The other genes were expressed heterogenously. Discussion: In patients with NLUTD associated with bladder fibrosis and/or failure to conservative treatment, the gene expression of factors related to increased extracellular matrix deposition and hypoxia appears to be relevant in the progression of bladder dysfunction. These findings are comparable with studies in animal models of bladder oulet obstrucion and hypoxia and reinforces the need for complentary studies to better understand the pathophysiology of the progression of bladder dysfunction in the NLTUD

As pointed out by I. Moerdijk and G. Reyes in [63], C&#8734-rings have been studied specially for their use in Singularity Theory and in order to construct topos models for Synthetic Differential Geometry. In this work, we follow a complementary trail, deepening our knowledge about them through a more pure bias, making use of Category Theory and accounting them from a logical-categorial viewpoint. We begin by giving a comprehensive systematization of the fundamental facts of the (equational) theory of C&#8734-rings, widespread here and there in the current literature - mostly without proof - which underly the theory of C&#8734-rings. Next we develop some topics of what we call a &#8734Commutative Algebra, expanding some partial results of [66] and [67]. We make a systematic study of von Neumann-regular C&#8734-rings (following [2]) and we present some interesting results about them, together with their (functorial) relationship with Boolean spaces. We study some sheaf theoretic notions on C&#8734-rings, such as &#8734(locally)-ringed spaces and the smooth Zariski site. Finally we describe classifying toposes for the (algebraic) theory of &#8734 rings, the (coherent) theory of local C&#8734-rings and the (algebraic) theory of von Neumann regular C&#8734-rings. / Conforme observado por I. Moerdijk e G. Reyes em [63], os anéis C&#8734 têm sido estudados especialmente tendo em vista suas aplicações em Teoria de Singularidades e para construir toposes que sirvam de modelos para a Geometria Diferencial Sintética. Neste trabalho, seguimos um caminho complementar, aprofundando nosso conhecimento sobre eles por um viés mais puro, fazendo uso da Teoria das Categorias e os analisando a partir de pontos de vista algébrico e lógico-categorial. Iniciamos o trabalho apresentando uma sistematização abrangente dos fatos fundamentais da teoria (equacional) dos anéis C&#8734, distribuídos aqui e ali na literatura atual - a maioria sem demonstrações - mas que servem de base para a teoria. Na sequência, desenvolvemos alguns tópicos do que denominamos Álgebra Comutativa C&#8734, expandindo resultados parciais de [66] e [67]. Realizamos um estudo sistemático dos anéis C&#8734 von Neumann-regulares - na linha do estudo algébrico realizado em [2]- e apresentamos alguns resultados interessantes a seu respeito, juntamente com sua relação (funtorial) com os espaços booleanos. Estudamos algumas noções pertinentes à Teoria de Feixes para anéis &#8734, tais como espaços (localmente) &#8734anelados e o sítio de Zariski liso. Finalmente, descrevemos toposes classicantes para a teoria (algébrica) dos anéis C&#8734, a teoria (coerente) dos anéis locais C&#8734 e a teoria (algébrica) dos anéis C&#8734 von Neumann regulares.

Introdução: As células do músculo liso vascular (CMLV) são quiescentes nos vasos adultos, com baixa capacidade de migração e de secreção de matriz extracelular, caracterizando fenótipo contrátil. Evidências apontam para a heterogeneidade fenotípica das CMLV ao longo da árvore arterial: há distribuição heterogênea de doenças e de resposta a determinadas drogas nos diferentes vasos, além de variabilidade de expressão dos genes de proteínas contráteis de músculo liso entre eles. O papel das CMLV, em fase adulta, é classicamente descrito como restrito à regulação do tônus de pequenos vasos, sendo insignificante a contribuição da mecânica das CMLV para a complacência das artérias elásticas. Existe a hipótese de que a viscoelasticidade das CMLV contribua para a mecânica final das artérias, sendo o enrijecimento dessas células associado à rigidez arterial. Objetivo: Estudar a variabilidade das propriedades mecânicas e de expressão proteica das CMLV, ao longo da árvore arterial, buscando identificar moduladores regionais para esse fenótipo. Avaliar se situações clínicas sabidamente associadas à rigidez arterial (envelhecimento, sexo feminino pós-menopausa, ancestralidade genética africana, diabetes mellitus e tabagismo) cursam com enrijecimento de CMLV. Métodos: 1) Estudou-se a composição e a organização da camada média de diferentes artérias. As CMLV desses vasos foram avaliadas quanto à viscoelasticidade de citoplasma (G), por meio do ensaio de Citometria Magnético Ótica de Oscilação e, quanto à expressão proteica global, usando cromatografia multidimensional e espectrometria de massas em tandem de alta resolução (Proteômica Shotgun). Os dados mecânicos obtidos foram correlacionados com as características da matriz extracelular (MEC) dos vasos de origem (porcentagem de elastina e quantidade de MEC). Em paralelo, foi realizado experimento de estiramento cíclico (10%/1Hz) das CMLV das diferentes artérias por 24 e 48h, seguido pela mensuração de rigidez de citoplasma. 2) Foram isoladas as CMLV de fragmentos de artéria mamária de 80 pacientes submetidos à cirurgia de revascularização do miocárdio, células essas que foram avaliadas quanto à viscoelasticidade de citoplasma (G, G\' e G\'\'). Elaborou-se modelo estatístico para avaliar se as variáveis clínicas idade, sexo feminino, ancestralidade africana, tabagismo e diabetes mellitus estavam associadas a alterações de mecânica celular. Resultados: 1) A viscoelasticidade das CMLV variou significativamente entre as artérias. As CMLV provenientes de artérias distais (artérias femoral e renal) mostraram-se significativamente mais rígidas que as CMLV de aorta torácica (p < 0,001). Identificou-se correlação negativa entre rigidez de CMLV e quantidade de MEC / elastina na camada média vascular. O regime de estiramento cíclico por 48h reduziu globalmente a rigidez das CMLV. As CMLV provenientes da aorta torácica expressaram maior quantidade de proteínas relacionadas com a estrutura e a organização do citoesqueleto em relação às CMLV da artéria femoral. 2) Constatou-se variabilidade interindividual de viscoelasticidade de CMLV e associação entre tabagismo e sexo feminino com enrijecimento de CMLV. Conclusões: As CMLV são heterogêneas quanto às propriedades mecânicas, à organização do citoesqueleto e à expressão proteica ao longo da árvore arterial, reforçando o conceito de plasticidade fenotípica das CMLV. A mecânica das CMLV é modulada pelas características da MEC e pela tensão circunferencial cíclica aplicada às paredes vasculares pelo fluxo sanguíneo. Mulheres pós-menopausa e tabagistas exibem enrijecimento de CMLV, sendo esse fato um provável contribuinte para a rigidez arterial associada a essas condições e um possível alvo terapêutico a ser avaliado futuramente / Rational: Vascular smooth muscle cells (VSMC) lose their ability to migrate and secrete extracellular matrix (ECM) with the end of vascular development, condition known as contractile phenotype and reversible in the presence of vascular injury. There is evidence of heterogeneity of VSMC phenotype along arterial tree, as the distribution of diseases (atherosclerosis) and the response to drugs vary between different vessels, as well as the expression of smooth muscle-contractile protein genes. The role played by VSMC mechanics on determining large arteries\' compliance was always considered irrelevant. It has been hypothesized that the VSMC mechanical properties are important for vascular mechanics, especially in the pathological scenario, where VSMC stiffening may be associated with arterial rigidity. Goals: Study the variation of VSMC mechanics and protein expression along arterial tree, identifying regional modulators of this phenotype. Evaluate if clinical situations associated with arterial rigidity (ageing, post-menopausal women, African ancestry, diabetes mellitus and smoking) concur with VSMC stiffening. Methods: 1) Different arteries were studied in terms of composition and organization of their media layer. VSMC isolated from these arteries were evaluated regarding cytoplasm viscoelasticity, measured using Optical Magnetic Twisting Cytometry Assay (OMTC), and protein expression, using two-dimensional liquid chromatography and tandem mass spectrometry (Shotgun Proteomics). Mechanical data were correlated with ECM characteristics (percentage of elastin and ECM amount) of the vessels of origin. In parallel, VSMC of different arteries were subjected to cyclic stretching (10%/1Hz) during 24 and 48h, followed by the measurement of their cytoplasm rigidity. 2) VSMC were isolated from fragments of mammary artery of 80 patients subjected to coronary bypass surgery and evaluated regarding their viscoelasticity (G, G\' e G\'\'). A statistic model was elaborated to address if the clinical variables age, female sex, African ancestry, smoking and diabetes mellitus were associated with changes of VSMC mechanics. Results: 1) VSMC viscoelasticity varied significantly amongst the studied arteries. VSMC from heart-distant arteries (femoral and renal arteries) were stiffer than VSMC from thoracic aorta (p < 0,001). There was a negative correlation between VSMC rigidity and the amount of ECM / percentage of elastin within the media layer. 48h-cyclic stretching was associated with a global reduction of VSMC rigidity. VSMC of thoracic aorta expressed significantly more proteins associated with cytoskeleton structure and organization than VSMC of femoral artery. 2) There was a significant inter-individual variation of VSMC viscoelasticity. Smoking and female sex were associated with VSMC stiffening. Conclusion: VSMC mechanics, cytoskeleton organization and protein expression are heterogeneous along arterial tree. VSMC mechanical properties are modulated by ECM characteristics and by regional mechanical forces. This reinforces the concept of phenotypic heterogeneity of VSMC. Post-menopausal women and smokers exhibit stiffer VSMC, representing an important factor for the understanding of the arterial rigidity associated with these conditions and also a possible future therapeutic target

As metaloproteinases, abundantes em venenos de serpentes da família Viperidae, apresentam homologia estrutural e funcional com as metaloproteinases de mamíferos (MMPs), cujos níveis estão elevados em doenças de natureza inflamatória, como a aterosclerose. A metaloproteinase BaP1, do veneno da serpente Bothrops asper, apresenta potente atividade inflamatória e constitui ferramenta científica importante para o estudo das ações das MMPs. Durante a aterosclerose, as células de músculo liso vascular (CMLVs) mudam do fenótipo contrátil para sintético, migram para a camada subendotelial do vaso, liberam mediadores inflamatórios e expressam MMPs. No entanto, o papel destas enzimas na resposta inflamatória das CMLVs e a potencial relação deste efeito com a migração das mesmas, não foi esclarecida. Neste estudo, investigou-se os efeitos da BaP1 em CMLVs, quanto à 1) indução da migração; 2) liberação de diferentes classes de mediadores inflamatórios e expressão de moléculas de adesão; 3) indução da mudança fenotípica das CMLVs; 4) expressão e participação de enzimas de síntese de prostaglandinas e de receptores de PGE2 na liberação deste eicosanoide; 5) participação de eicosanoides e da IL-1&#946 na migração e mudança de fenótipo das CMLVs. Os resultados obtidos, a partir dos ensaios de transwell e wound healing, mostraram que a BaP1(50nM) induziu a migração das CMLVs, após 48 h, mas não a proliferação celular, observada pelo ensaio de ciclo celular. Além disso, a metaloproteinase induziu aumento da liberação de PGE2 (1-48h), LTB4 (1-3h), IL-1&#946 (12-24h), MCP-1 (24-48h) e fractalcina (24-48h), mas não de PGI2 e nem TXA2, analisados por ensaios de EIA e multiplex. Ainda, a BaP1 aumentou a expressão proteica de COX-2 (1° h) e de PGESm-1 (4° h), analisada por Western blotting, e expressão gênica das sFLA2-IIA (30 min) e cFLA2-IVA (30 min), verificada por PCR em tempo real, sem alterar os níveis de COX-1, dos receptores EP1, EP2, EP3 e EP4, de ICAM-1 e VCAM-1 e da iFLA2. A intervenção farmacológica com os inibidores de COX-2 ou de FLA2 intracelulares reduziu a liberação de PGE2 induzida pela BaP1. Além disso, o pré-tratamento das células com o inibidor da FLAP e com os antagonistas do receptor de IL-1&#946 ou do receptor EP3 reduziu a migração celular induzida pela BaP1. Esta metaloproteinase também induziu a mudança de fenótipo contrátil para o sintético, das CMLVs, verificada pela diminuição da expressão de &#945-actina pelo ensaio de citometria de fluxo. A inibição da COX-2 e da FLAP não alterou este efeito. Este conjunto de dados demonstra a capacidade da BaP1 estimular diretamente as CMLVs para migração, liberação de mediadores inflamatórios e a expressão de COX-2, PGESm-1, sFLA2-IIA e cFLA2-IVA. A produção de PGE2 induzida pela BaP1 depende das FLA2s intracelulares, com ativação das vias da COX-1 e -2. A migração das CMLVs, induzida pela BaP1, depende da PGE2 via ativação do receptor EP3, do LTB4 e da IL-1&#946. Ainda, esta metaloproteinase estimula a mudança fenotípica das CMLVs para o estágio sintético, em que as CMLVs migram e proliferam. Os dados deste estudo, ao demonstrarem que as metaloproteinases contribuem para o desenvolvimento de eventos inflamatórios, em CMLVs, apontam um papel adicional desta classe de enzimas em doenças de natureza inflamatória, como a aterosclerose. / Metalloproteinases are abundant enzymes in Viperidae family snake venoms and exhibit structural and functional homology with mammalian matrix metalloproteinases (MMPs). The levels of these enzymes are incresead in inflammatory diseases, such as atherosclerosis. The BaP1 metalloproteinase from Bothrops asper snake venom presents potent inflammatory activity and constitutes important scientific tool for the study of the actions of MMPs. During atherosclerosis, vascular smooth muscle cells (VSMCs) switch their phenotype from a contractile to a synthetic state, migrate into the subendothelial vessel layer, release inflammatory mediators and express high levels of MMPs. However, the role of these enzymes in the inflammatory response of VSMCs and the potential relationship of this effect with cell migration have not been clarified. In this study, we investigated the effects of BaP1 on CMLVs with focus on: 1) induction of cell migration; 2) release of different classes of inflammatory mediators and protein expression of adhesion molecules; 3) induction of VSMCs phenotype switching; 4) expression and participation of prostaglandin synthesis enzymes and PGE2 receptors in the release of this eicosanoid; 5) participation of eicosanoids and IL-1&#946 in migration and phenotype switching of VSMCs. Results obtained from the transwell and wound healing assays showed that BaP1 (50nM) induced VSMCs migration after 48 h, but not cell proliferation, observed by the cell cycle assay. In addition, this metalloproteinase caused release of PGE2 (1-48h), LTB4 (1-3h), IL-1 (12-24h), MCP-1 (24-48h) and fractalkine (24-48h), but not PGI2 and TXA2, analyzed by EIA and multiplex assays. Furthermore, BaP1 increased protein expression of COX-2 (1 h) and PGESm-1 (4 h), analyzed by western blotting and gene expression of sFLA2-IIA (30 min) and cFLA2-IVA (30 min), evaluated by real-time PCR, without altering COX-1, EP1, EP2, EP3 and EP4, ICAM-1 and VCAM-1 and iFLA2 levels. Pharmacological intervention with COX-2 or intracellular FLA2 inhibitors reduced PGE2 release induced by BaP1. In addition, pretreatment of cells with either a FLAP inhibitor, or IL-1&#946 receptor, or EP3 receptor antagonist reduced cell migration induced by BaP1. This metalloproteinase also induced conversion of contractile VSMCs to an synthetic phenotype, as evidenced by decrease of -actin expression, analyzed by flow cytometry assay. Inhibition of COX-2 and FLAP did not alter this effect. Altogether, these data demonstrate the ability of BaP1 to directly stimulate VSMCs for migration, release of inflammatory mediators and expression of COX-2, PGESm-1, sFLA2-IIA and cFLA2-IVA. PGE2 production induced by BaP1 depends on the intracellular FLA2s, with activation of COX-1 and -2 pathways. VSMCs migration induced by BaP1 depends on PGE2 via EP3 receptor engagement, LTB4 and IL-1&#946. Furthermore, this metalloproteinase stimulates VSMCs phenotypic switching to a synthetic phenotype, in which these cells migrate and proliferate. These data demonstrate that metalloproteinases can contribute to the development of inflammatory events in VSMCs, evidencing an additional role of this class of enzymes in inflammatory diseases, such as atherosclerosis.

Une des propriétés majeures de la thrombine est le caractère pléiotropique de ses effets physiologiques et pathologiques, à la fois dans le compartiment sanguin et tissulaire de la paroi. Notre hypothèse est que les changements phénotypiques des cellules musculaires lisses vasculaires (CMLVs) participent aux modifications des propriétés pro- et anticoagulantes de la paroi. Les objectifs ont été d’étudier : (i) le rôle prothrombotique des CMLVs dans l’hypertension chez le rat SHR et le syndrome métabolique (Smet) chez le rat Zucker, (ii) les mécanismes de régulation de la génération de thrombine par l’intégrine αvβ3 des CMLVs (récepteur de la pro- thrombine), et de développer des glyco-peptides fluorés pour l’imagerie permettant d’évaluer l’activité de cette intégrine dans la paroi, et (iii) d’évaluer l’effet de variants génétiques du locus 9p21 de susceptibilité aux maladies coronariennes sur le phénotype de coagulation. Résultats : Les CMLVs sont responsables du phénotype prothrombotique de la paroi artérielle associée à l’hypertension chez le rat SHR. Les acides gras libres et l’inflammation vasculaire augmentent la génération de thrombine dans les 2 compartiments ce qui se traduit par une fibrinolyse diminuée et une activité métallo-protéinase augmentée chez Le rat Zucker. L’invalidation de l’intégrine αvβ3 des CMLVs diminue la génération de thrombine dans les 2 compartiments et ralentit la survenue de thrombose carotidienne en réponse à une stimulation l’angiotensine. Le traçage de l’intégrine αvβ3 par des glyco-peptides comprenant une séquence RGD a été validé au niveau plaquettaire et des CMLVs. La souris invalidée pour le locus 9p21 exprime un phénotype pro-thrombotique qui est retrouvé chez l’homme pour certains variants (rs10120688 et rs1333040) dans ce locus. En conclusion, la CML est un support cellulaire clé de réactions procoagulants et pourrait être impliqué via les intégrines et/ou ses récepteurs pour la thrombine dans un couplage thrombine tissulaire – rigidité cellulaire dans les pathologies vasculaires / One of the major properties of thrombin is the pleiotropic character of its physiological and pathological effects, both in the blood compartment and the tissue of the arterial wall. We hypothesized that the phenotypic changes of vascular smooth muscle cells (VSMCs) are involved in modifications of pro- and anti-coagulant properties of the arterial wall. The objectives were to examine: (i) the prothrombotic role of VSMCs in hypertension of SHR rats and in the metabolic syndrome (Smet) of Zucker rats, (ii) regulatory mechanisms of thrombin generation by integrin αvβ3 of VSMCs (a pro-thrombin receptor), and to develop fluorinated glyco- peptides for imaging, to assess the activity of this integrin in the wall, and (iii) evaluate the effect of genetic variants of the 9p21 locus that give a susceptibility to coronary heart disease on the coagulation phenotype. Results: The VSMCs are responsible for the prothrombotic phenotype of the arterial wall associated with hypertension in SHR rats. Free fatty acids and vascular inflammation increase thrombin generation in the two compartments resulting in decreased fibrinolysis and an increased metallo-proteinase activity in the Zucker rats. The invalidation of integrin αvβ3 of VSMCs reduced thrombin generation in the two compartments and slowed angiotensin-induced carotid thrombosis. Tracing of the integrin αvβ3 by glyco-peptides including RGD was validated at the platelet level and VSMCs. Mice invalidated for the 9p21 locus express a prothrombotic phenotype that is found in humans for certain variants (rs10120688 and rs1333040) in this locus. In conclusion, the VSMC is a cell supported key to procoagulant reactions and may be involved via integrins and/or its receptors for thrombin in the ”tissular thrombin - cell rigidity” coupling in vascular pathologies

Les propriétés du béton constituant les enceintes de confinement des centrales électronucléaires évoluent sous les effets de mécanismes de vieillissement résultant notamment de transferts couplés de chaleur et de masse au sein du matériau. Ces phénomènes peuvent être modélisés par des équations de transports moyennées : lois de Fick pour le transport d’espèces en solution et lois de Fourier pour la description de la diffusion thermique. Dans cette étude, les développements concernent la diffusion de la thermique dans un milieu hétérogène fissuré représentant un matériau cimentaire dégradé chimiquement. Le problème thermo-mécanique est traité à l'aide d'une approche multi-corps reliés par des lois d’interactions enrichies (zones cohésives). La diffusion thermique est écrite dans le formalisme cohésif-volumique en prenant en compte le couplage entre un état d'endommagement local de la zone cohésive et une conductivité homogénéisée. Afin d'optimiser les coûts de calculs, une étude est menée sur la dimension d'un volume élémentaire représentatif (VER). Pour cela, la méthode d'eigenerosion est étendue à la fissuration de milieux hétérogènes puis appliquée aux milieux cimentaires. La propagation de fissures sous chargement thermique est ensuite analysée dans des VERs de béton dégradés représentatifs des enceintes de confinement des centrales nucléaires après plusieurs années. Le vieillissement est modélisé par un taux de pré-dégradation initial entre le mortier et les granulats. Le développement de multi-fissures est relié au taux de pré-dégradation et la formation "d'écrans" à la diffusion de la thermique est mise en avant. / During their confinement in a nuclear power plant, the mechanical properties of the constitutive materials of concrete change as a result of ageing. This is due to the transportation of chemical species at the microscopic level of the media. Firstly, this can be modelled with average equations. The Fick laws represent the evolution of chemical diffusion and the Fourier laws, the transportation of heat at a mesoscopic level. In this research, we will consider thermal evolution on a fractured media.This thermomechanical problem is solved with a staggered method. The mechanical contribution used an approach based on multi-bodies system linked with cohesive zone models. The thermal problem is based on the approximation of the heat transfer equation at the cohesive interface. This approach has been implemented and validated. The description of the heat trough the interface is composed with the definition of an homogenised conductivity and the local damage parameter. In order to optimize the computational cost with a good agreement of the crack propagation, a criterion is proposed for sizing a representative elementary volume (REV). The eigenerosion method is used, validated and extended to heterogeneous media. Two studies are carried out on the morphological properties on a cementious media. As a result of those studies, a minimal size for a REV is defined.Crack spread under thermal loads are investigated on a media representing the concrete of the containment of a nuclear power station. The ageing effect are taken into account as an initial damage between the mortar and the aggregates. These parameters are expressed in terms of rate of initial damage. A study is proposed for different values of this rate. As assumed, the development of multi-cracks is linked with the rate of initial damage and the creation of thermal border is proposed.

No description available.

As pointed out by I. Moerdijk and G. Reyes in [63], C&#8734-rings have been studied specially for their use in Singularity Theory and in order to construct topos models for Synthetic Differential Geometry. In this work, we follow a complementary trail, deepening our knowledge about them through a more pure bias, making use of Category Theory and accounting them from a logical-categorial viewpoint. We begin by giving a comprehensive systematization of the fundamental facts of the (equational) theory of C&#8734-rings, widespread here and there in the current literature - mostly without proof - which underly the theory of C&#8734-rings. Next we develop some topics of what we call a &#8734Commutative Algebra, expanding some partial results of [66] and [67]. We make a systematic study of von Neumann-regular C&#8734-rings (following [2]) and we present some interesting results about them, together with their (functorial) relationship with Boolean spaces. We study some sheaf theoretic notions on C&#8734-rings, such as &#8734(locally)-ringed spaces and the smooth Zariski site. Finally we describe classifying toposes for the (algebraic) theory of &#8734 rings, the (coherent) theory of local C&#8734-rings and the (algebraic) theory of von Neumann regular C&#8734-rings. / Conforme observado por I. Moerdijk e G. Reyes em [63], os anéis C&#8734 têm sido estudados especialmente tendo em vista suas aplicações em Teoria de Singularidades e para construir toposes que sirvam de modelos para a Geometria Diferencial Sintética. Neste trabalho, seguimos um caminho complementar, aprofundando nosso conhecimento sobre eles por um viés mais puro, fazendo uso da Teoria das Categorias e os analisando a partir de pontos de vista algébrico e lógico-categorial. Iniciamos o trabalho apresentando uma sistematização abrangente dos fatos fundamentais da teoria (equacional) dos anéis C&#8734, distribuídos aqui e ali na literatura atual - a maioria sem demonstrações - mas que servem de base para a teoria. Na sequência, desenvolvemos alguns tópicos do que denominamos Álgebra Comutativa C&#8734, expandindo resultados parciais de [66] e [67]. Realizamos um estudo sistemático dos anéis C&#8734 von Neumann-regulares - na linha do estudo algébrico realizado em [2]- e apresentamos alguns resultados interessantes a seu respeito, juntamente com sua relação (funtorial) com os espaços booleanos. Estudamos algumas noções pertinentes à Teoria de Feixes para anéis &#8734, tais como espaços (localmente) &#8734anelados e o sítio de Zariski liso. Finalmente, descrevemos toposes classicantes para a teoria (algébrica) dos anéis C&#8734, a teoria (coerente) dos anéis locais C&#8734 e a teoria (algébrica) dos anéis C&#8734 von Neumann regulares.

To this day heart failure with preserved ejection fraction (HFpEF) remains a poorly understood malady. Half of all heart failure (HF) cases are HFpEF, and the prevalence of HF is on the rise. Unlike HF with reduced ejection fraction, HFpEF has no treatment options and is often times difficult to diagnose because victims of HFpEF often have pre-existing conditions. Vascular endothelial growth factor (VEGF) has been implicated in maintaining myocardial health and is thought to play a role in HFpEF. We sought to test the hypothesis that VEGF-A plays a role in HFpEF in a hypertensive murine model of HFpEF. Using Western blot analysis we found that there was an up regulation of VEGF-A in the homogenized left ventricle (LV) of our HFpEF mice. Unexpectedly, there was a down regulation of VEGF-A in the homogenized tissue from the aorta in those mice. To study the circulating levels of VEGF in our HFpEF mice we used an ELISA. We found that our HFpEF mice had similar levels of circulating VEGF as our control. This suggests that VEGF has paracrine/autocrine role in our HFpEF model rather than endocrine, like our human data suggested. To identify the cells responsible for the expression profile we saw in the homogenized tissue data we looked at the response of adult rat ventricular myocytes (ARVM) and vascular smooth muscle cells (VSMC) to aldosterone stimulation at short (1hr) and long (24hr) time points at both physiological (50nm) and pathological (1μm) concentrations. To do this analysis we recruited the help of Western blot, ELISA and RT-PCR techniques to construct a consistent VEGF expression profile. The Western blot ARVM data showed statistically significant (P<0.05) increase in VEGF-A to pathological doses of aldosterone, especially at the longer time point. When we tested the VSMC using Western blot analysis, we found that the trend of our n=1 sample suggested a strong response to the physiological dose of aldosterone in the short term. Using the more sensitive ELISA technique to measure the VEGF content of our VCMS we increasing our sample size to n=4 and found no statistically significant (p=NS) response to aldosterone stimulation from the VSMC. However, looking at the trends in the data it is clear that VSMC increases VEGF in response to long-term physiological doses of aldosterone. This is contrary to what we found using Western blot analysis, so we queried the VEGF mRNA from the VSMC to settle the score. Unfortunately, this too proved fruitless. The RT-PCR data was not significant and the trend was that of the ARVM expression profile. We initially turned to VSMC because we hypothesized that they could contribute to the paracrine/autocrine activity similar to what we saw in the LV from the ARVM. It is unclear if VSMC play a role in HFpEF progression, but their lack of consistent response to aldosterone could potential explain the down regulation of VEGF-A we observed in the aorta of our HFpEF mice. We initially sough to test the hypothesis that VEGF-A plays a role in our HFpEF mouse model, what we found was that ARVM contribute to localized VEGF-A increased production in the LV while in the aorta there is a down regulation of VEGF-A in our HFpEF model, we are unable to make any conclusion about VSMC response to aldosterone because of insufficient sample size. Thus in conclusion, it appears that VEGF-A does play a role in our HFpEF model specifically in a paracrine/autocrine manner in the LV where the ARVM contributes to the increased production of the cytokine.

L’objectif de cette thèse était de caractériser le rôle des différentes familles de phosphodiestérases (PDEs), les enzymes de dégradation du 3'-5'- adénosine monophosphate cyclique (AMPc), dans la régulation de la signalisation de l’AMPc dans deux types de cellules musculaires lisses (CMLs), l’aorte de rat (CMLAR) et la vessie de rat néonatal (CMLVRN). Dans les CMLARs en culture, nous avons déterminé le profil d’expression et d’activité des PDE-AMPc. Nous avons alors montré, à l’aide de la technique de FRET basée sur une sonde sensible à l’AMPc pour mesurer l’AMPc en temps réel dans une cellule isolée, que l’inhibition de la PDE4 démasque un effet d’hydrolyse de l’AMPc cytosolique par la PDE1 et la PDE3, alors que les PDE3 et PDE4 agissent de façon synergistique dans le compartiment sous-membranaire. Les mécanismes de cette compartimentation subcellulaire des signaux restent à caractériser.Dans les CMLVRNs, les PDE3 et PDE4 régulent les contractions phasiques, par des mécanismes différents. L’inhibition de la PDE4 limite les contractions stimulées par le carbachol par un mécanisme dépendant de la protéine kinase A, impliquant une augmentation de la fréquence des sparks calciques, qui entrainent l’activation des canaux potassiques BK, assurant en final une diminution des transitoires calciques. Au contraire, l’effet de l’inhibition de la PDE3 implique la protéine kinase G mais par un mécanisme qui reste à définir.En conclusion, ce travail montre que dans les CMLs, les différents familles de PDE-AMPc sont douées de spécificité de fonction et/ou de mécanisme d’action, et participent ainsi à une compartimentation subcellulaire des voies de signalisation. / The aim of the present thesis was to characterize the role of the different families of phosphodiesterases (PDEs), the enzymes degrading 3'-5'-cyclic adenosine monophosphate (cAMP), in controlling the cAMP signalling in two distinct smooth muscle cells (SMCs), the rat aorta SMC (RASMCs) and the rat bladder SMC (RBSMCs).In cultured RASMCs, we firstly characterized the pattern of cAMP-PDE expression and activity. We then showed, by using a FRET-based cAMP sensor to explore cAMP signals in living cells, that PDE4 inhibition unmasks an effect of PDE1 and PDE3 on cytosolic cAMP hydrolyzis, whereas PDE3 and PDE4 act synergistically at the submembrane compartment. The mechanisms of this subcellular compartmentation need to be characterized. In neonatal RBSMCs, we showed that both PDE3 and PDE4 are involved in regulating the phasic contractions albeit through distinct mechanisms. PDE4 inhibition inhibits the carbachol-enhanced contractions through a protein kinase A-dependent pathway involving an increase in Ca2+ sparks frequency which activates BK channels to ultimately decrease Ca2+ transients, whereas PDE3 inhibition acts through a protein kinase G-dependent pathway through a still unknown mechanism.In conclusion, our work shows that in the SMC, the different cAMP-PDE families exhibit a specificity in their function and/or mechanism of action, thus participating to a subcellular signaling compartmentation.