Os mecanismos pelos quais os agentes infecciosos, independentes ou não de meio ambiente permissivo, podem promover a aterogênese e as manifestações clínicas não estão completamente esclarecidos. Apesar das numerosas publicações demonstrando a presença de antígenos ou DNA de agentes infecciosos nas placas de ateroma, a questão se o agente infeccioso pode iniciar o processo aterosclerótico ou agravá-lo permanece sem resposta, possibilitando o aprofundamento das pesquisas neste assunto. Desta forma, este trabalho tem como objetivo estudar se a infecção experimental, por C.pneumoniae e/ou M.pneumoniae, em camundongos C57BL/6 apoE KO induziria ou afetaria a intensidade e a característica de vulnerabilidade da placa ateromatosa, de acordo com o sexo e/ou a dieta rica em colesterol. Métodos: um grupo de camundongos recebeu dieta enriquecida com 1% de colesterol (hiperlipidêmica), e o outro ração com formulação adequada para espécie (normolipidêmica), desde os dois meses de idade. Aos 8 meses de idade foram subdivididos, inoculados com 106 UFI de C.pneumoniae e/ou 106 UFC de M.pneumoniae, por via intraperitoneal, reinoculados um mês após e sacrificados aos 10 meses de idade. Para análise histopatológica secções transversais das aortas torácicas foram processadas para emblocamento em parafina, cortadas com 5 µm de espessura e coradas pelas técnicas de hematoxilina-eosina e tricrômico de Masson. As medianas das variáveis: altura da placa, área da placa, área de gordura da placa, área da artéria, área da luz e porcentagem de obstrução da luz da artéria dos diferentes grupos foram submetidas ao teste de Mann Whitney, com o nível de rejeição de 5%. Resultados: a infecção por C.pneumoniae e/ou M.pneumoniae causou agravamento da aterosclerose tanto em camundongos C57BL/6 apoE KO machos quanto em fêmeas. No entanto, as fêmeas infectadas somente com M.pneumoniae evoluíram com placas mais instáveis, representadas por maior remodelamento positivo. A co-infecção por C.pneumoniae e M.pneumoniae induziu placas mais estáveis, ou seja, com menor conteúdo de gordura e sem remodelamento, tanto nos machos quanto nas fêmeas. A introdução de dieta rica em colesterol levou ao não desenvolvimento de remodelamento positivo do vaso nas fêmeas infectadas por M.pneumoniae, mas sim nas co-infectadas por C.pneumoniae e M.pneumoniae que apresentaram placas mais instáveis, por serem mais volumosas e com maior conteúdo de gordura. Nos machos houve desenvolvimento de placas mais gordurosas nos infectados por C.pneumoniae. Conclusão: A infecção por C.pneumoniae e/ou M.pneumoniae em camundongos C57BL/6 apoE KO levou ao desenvolvimento ou agravamento de placas de aterosclerose, com diferenças em relação a intensidade e padrões de vulnerabilidade de acordo com o sexo versus o tipo de agente infecciosos. Os subgrupos infectados de fêmeas apresentaram maior agravamento da aterosclerose do que os machos. A dieta rica em colesterol agravou a intensidade da aterosclerose e mudou os padrões de vulnerabilidade dos subgrupos infectados. / Independent of the presence or not a favorable ambient, mechanisms by which infectious agents may boost atherogenesis and clinical aspects are not fully elucidated. In spite of many demonstrations of infeccious agent antigens or DNA, the question if the infection may iniciate or aggravate the atherosclerotic process remains unanswered, requiring further studies. Therefore, the present work studies if the experimental infection of C57BL/6 apoE KO mice by C. pneumoniae and/or M. pneumoniae induces or affects the intensity of atherosclerosis and its characteristics of plaque vulnerability, with regard to the gender and/or the cholesterolrich diet. Methods: a group of mice was fed with 1% cholesterol-enriched diet (hyperlipidemic), from two months of age; the other group received adequately formulated food for the species (normolipidemic). At eight months of age, the mice were subgrouped according to inoculation intra-peritoneally with 106 IFU of C. pneumoniae and/or 106 CFU of M. pneumoniae, and re-inoculation one month later. They were killed at ten months of age. Cross-sectional thoracic aorta fragments were studied in embedded in paraffin block sections stained Hematoxylin-eosin and Masson?s trichromic techniques. Differences in the median of the variables: plaque height, plaque area, area of fat plaque, luminal area and percent obstruction of the lumen searched using the Mann Whitney?s test, with a 5% level of rejection. Results: the infection by C. pneumoniae and/or M. pneumoniae worsened atherosclerosis in both males and females C57BL/6 apoE KO mice. However, the M. pneumoniae inoculated female group presented more unstable plaques represented by positive remodeling of the vessel. The co-infection by both bacteria induced more stable plaque represented by low fat content and absence of vessel remodeling, in both male and female mice. The introduction of cholesterol enriched diet led to lack of positive vessel remodeling in M.pneumoniae inoculated female group, but development of unstable plaques characterized by large plaque are with high content of fat, in co-infected ones. In male groups there was development of plaque with higher fat content in the subgroup inoculated with C.pneumoniae. Conclusion: Infection by M. pneumoniae and/or C. pneumoniae, in C57BL/6 apoE KO mice, led to development or aggravation of atherosclerotic plaques, with differences regarding intensity and pattern of vulnerability according to the gender versus type of infectious agents. The infected female groups presented more aggravation of atherosclerosis than male ones. Cholesterol enriched diet aggravated the intensity of atherosclerosis and changed the patterns of vulnerability of infected subgroups.

La prévalence de la maladie asthmatique est en constante augmentation à travers le monde. Au cours de la vie dun asthmatique, la fonction pulmonaire diminue plus fortement comparativement aux sujets normaux. Ceci est lié à linflammation et au remodelage des voies aériennes. Les caractéristiques principales du remodelage bronchique consistent en des dommages épithéliaux, une hyperplasie des cellules musculaires lisses, une hyperplasie glandulaire, et une fibrose sous-épithéliale incluant un dépôt de collagène dans la lamina reticularis de lépithélium bronchique. Les traitements actuels de lasthme ne sont pas efficaces dans le contrôle du remodelage bronchique et ne permettent pas de contrôler efficacement la maladie dun point de vue clinique. Pour cette raison, le développement de nouveaux agents thérapeutiques semble nécessaire afin de permettre un contrôle de linflammation aigue, de lhyperréactivité et du remodelage bronchiques. Dans ce travail, nous avons dans un premier temps, mis au point un modèle murin permettant létude de la réponse inflammatoire aigue mais également du remodelage bronchique. Grâce au développement de ce modèle murin, nous avons pu détecter par analyse transcriptomique de nouvelles cibles thérapeutiques potentielles pour la maladie asthmatique telles que lAgr2, le Cd209e, le Col6 et le C1qa. Dautres gènes préalablement décrits comme surexprimés dans lasthme ont également été étudiés afin de valider notre modèle (MMP-12, Fcgr2b, Ccl8, chi3l3 et Arg1). Suite à cela, nous avons décidé dinvestiguer le rôle de la MMP-12 dans la pathologie asthmatique car bien que ce gène soit connu pour être surexprimé dans la maladie, son rôle exact dans la pathologie asthmatique nétait pas encore clair. La MMP-12, également appelée métalloélastase du macrophage, est une pro-enzyme de 54 kDa qui est clivée en deux formes actives de 45 kDa et de 22 kDa. Elle est exprimée principalement par les macrophages alvéolaires mais également par les cellules épithéliales bronchiques, les cellules dendritiques et les cellules musculaires lisses. Le substrat principal de cette enzyme est lélastine qui représente environ 2.5% du poids sec des poumons. En plus de son activité élastolytique, la MMP-12 peut dégrader dautres composants de la matrice extracellulaire comme le collagène de type IV, la fibronectine, la laminine, la vitronectine, les protéoglycans, etc. Elle a également la capacité dactiver la pro-MMP-2 et la pro-MMP-3 qui peuvent à leur tour activer la pro-MMP-9 et la pro-MMP-1. Cette cascade protéolytique explique comment la MMP-12 peut provoquer une dégradation exagérée dune grande variété de composant de la matrice extracellulaire. La MMP-12 est également capable de dégrader dautres substrats non liés à la matrice extracellulaire. Une régulation anormale de lexpression de la MMP-12 est rencontrée dans différentes pathologies telles que les maladies inflammatoires de la peau, lanévrisme de laorte, le cancer et lemphysème pulmonaire. Afin détudier plus avant le rôle de la MMP-12 dans linflammation allergique et dans le remodelage bronchique, nous avons appliqué notre modèle murin à des souris déficientes pour le gène de la MMP-12 ainsi quaux souris wild-type correspondantes. A la fin de chaque protocole expérimental, la résistance bronchique mesurée à laide du système Flexivent est moins importante chez les souris déficientes pour le gène de la MMP-12 exposées à lallergène lorsquon les compare aux souris de type sauvage correspondantes. Les pourcentages déosinophiles présents dans le lavage bronchoalvéolaire des souris déficientes pour le gène de la MMP-12 exposées à lallergène sont significativement moins importants que ceux présents chez les souris de type sauvage correspondantes et ce quelque soit le temps dexposition. Les mêmes différences ont été observées pour le score inflammatoire et pour linfiltration des éosinophiles au sein des parois bronchiques. Le pourcentage de cellules caliciformes présentes dans lépithélium bronchique est significativement moins important chez les souris déficientes pour le gène de la MMP-12 exposées à lallergène durant 1 et 5 semaines comparativement aux souris de type sauvage correspondantes mais plus aucune différence na pu être observée après 10 semaines dexposition. Le nombre de mastocyte est significativement moins important chez les souris déficientes pour le gène de la MMP-12 exposées à lallergène durant 5 et 10 semaines comparativement aux souris sauvages correspondantes. Aucune différence na pu être observée après 1 semaine dexposition. Enfin, lépaisseur de la couche de muscle lisse est significativement moins importante chez les souris déficientes pour le gène de la MMP-12 exposées à lovalbumine durant 5 et 10 semaines comparativement aux souris de type sauvage correspondantes. Suite à ces résultats, nous avons montré que le MMP-12 était une cible thérapeutique potentielle pour lasthme. Nous avons également montré que les principales modifications dues au déficit de ce gène survenaient dès la première semaine dexposition. Nous avons donc décidé de tester les effets de linstillation dun siRNA ciblant la MMP-12 dans notre modèle murin dinflammation aigue. Un western blot et une PCR spécifiques de la MMP-12 ont été réalisés afin de sassurer que le siRNA permettait bien de diminuer lexpression de ce gène. Linstillation de siRNA ciblant la MMP-12 entraine bien une diminution de lexpression de ce gène tant au niveau de lARNm quau niveau protéique. Le pourcentage déosinophiles présents dans le lavage bronchoalvéolaire des souris exposées à lallergène ayant été instillées avec le siRNA ciblant la MMP-12 est significativement diminué par rapport à celui des souris exposées à lallergène ayant été instillées avec du PBS ou avec un siRNA scramble. De plus, cette diminution dépend de la dose de siRNA utilisée. Les mêmes observations ont pu être réalisées en ce qui concerne linfiltration des éosinophiles au sein des parois bronchiques ainsi que pour le pourcentage de cellules à mucus présentes dans lépithélium bronchique. Linstillation du siRNA chez les souris exposées à lallergène entraine une diminution du score inflammatoire par rapport aux souris ayant été instillées avec du PBS ou avec le siRNA scramble mais cette diminution semble moins dépendre de la dose de siRNA utilisé. Les niveaux dexpression de CCL-11 et de lIL-13 mesurés par ELISA sont diminués suite au traitement avec le siRNA ciblant lARNm de la MMP-12 comparativement aux souris contrôles. De même, les niveaux dexpression du TNF-α et de larginase I mesurés par western blot sont diminués après linstillation du siRNA ciblant la MMP-12. Au terme de ce travail, nous pouvons donc conclure que le MMP-12 joue un rôle important dans la pathologie asthmatique et que ladministration de siRNA ciblant ce gène permet de diminuer linflammation à éosinophiles ainsi que lhyperplasie des cellules à mucus. Ceci permet de penser que la MMP-12 est une nouvelle cible thérapeutique potentielle pour le traitement de lasthme.

L'électroconvulsivothérapie (ECT) est une réponse thérapeutique reconnue comme efficace face à la résistance et au délai d'action des agents pharmacologiques, pour les épisodes dépressifs majeurs. Bien que l'efficacité thérapeutique des ECT soit reconnue de tous, la connaissance incomplète des effets biologiques qui sous tendent l'effet thérapeutique contribue à discréditer ce soin. Les marqueurs biologiques ne sont pas facilement accessible chez l’Homme, il a donc été développé chez les rongeurs, l’équivalent des ECT : les stimulations électro-convulsives (ECS). A ce jour, les divers résultats provenant des études chez les animaux n’ont permis d’obtenir des conclusions claires, et ceci est majoritairement dû à l’utilisation d’animaux naïfs. Dans ce contexte, l'objectif du projet de thèse était d'analyser les effets biologiques et comportementaux des ECS répétées, modèle de l'électro convulsivothérapie, sur un modèle animal de troubles psychiatriques : la souris MAP6 KO. En effet, ces souris expriment constitutivement des altérations biologiques et comportementales assimilées à certains symptômes de la dépression. Nous avons montré qu’un traitement par ECS améliore certaines des altérations comportementales des souris MAP6 KO, avec un maintien dans le temps différent selon le comportement analysé. Au niveau biologique, le traitement par ECS n’induit pas de modification détectable de l’expression des protéines dans le cerveau, mais induit une augmentation de la neurogénèse adulte hippocampal, et plus particulièrement une augmentation du taux d'intégration des neurones néoformés. L'augmentation du nombre de nouveaux neurones survivants pourrait être favorisée par l’augmentation de la complexité de l’arborisation dendritique et de la densité synaptique. De plus nous observons une augmentation de la densité des épines dendritiques dans les neurones corticaux et une augmentation de l’expression du BDNF dans l’hippocampe. En conclusion, le projet de thèse montre que les ECS appliquées aux souris MAP6 KO-ont un effet important au niveau de la plasticité cérébrale et plus particulièrement au niveau de l’intégration des nouveaux neurones issus de la neurogénèse adulte hippocampique. / In Major depressive disorder, the electroconvulsive therapy (ECT) is recognized as an effective treatment to face drug-resistance and action latency of pharmacological agents. Although the therapeutic efficacy of ECT is internationally recognized, the ill-characterized biological effects of ECT contribute to discredit this treatment. Direct access of specific markers in human is not easy, prompting the development of the animal counterpart of ECT, the electro convulsive stimulations (ECS). To date, current results arising from ECS studies in animals remain not fully conclusive because mainly data have been obtained with unchallenged/naive animals. The objective of this project was to analyze the biological and behavioral effects of ECS treatment, on an animal model of psychiatric disorder: the MAP6-KO mice. Indeed MAP6-KO mice (also known as STOP KO mice) constitutively exhibit behavioural and biological features relevant to some aspects of major depressive disorder. We found that ECS treatment has an overall beneficial effect on several constitutive behavioural defects, displayed by MAP6 KO mice, with variable lasting times. At biological levels, ECS treatment did not induce detectable modification of brain proteins expression pattern, but induce an overall increase of hippocampal neurogenesis, and more particularly potentiate the survival rate of newborn neurons, probably through an increase dendritic complexity and dendritic spines density. We also found that ECS enhance dendritic spines density in vivo in cortical neurons and increase hippocampal levels of the trophic factor, BDNF. In summary our work provide evidence that ECS treatment when applied to MAP6 KO mice induces major neuronal plasticity events, the stronger being the increased integration rate of hippocampal new-born neurons.

Os leucotrienos (LTs) são mediadores inflamatórios derivados da via 5- lipoxigenase (5-LO), com contribuição relevante na reabsorção óssea. Neste estudo investigamos o papel dos LTs na diferenciação osteogênica e o seu impacto na osteoclatogênese. Assim, foi avaliado o perfil ósseo dos camundongos 129/Sv (WT) e 5-LO Knockout (5-LO KO) por meio de microtomografia computadorizada, evidenciando maior densidade óssea vertebral e trabéculas mais espessas em machos 5-LO KO. Após isso, osteoblastos primários (OBL) foram isolados e cultivados para determinar a atividade de fosfatase alcalina (ALP) e o potencial de mineralização. Resultados mostraram que OBL KO possui maior atividade de ALP e mineralização, em todos os períodos quando comparados com WT. Em adição, o tratamento com os LTs B4 e D4 inibiu a deposição de cálcio. Os inibidores da síntese de LTs e os antagonistas do BLT1/2 foram efetivos em recuperar a formação dos nódulos mineralizados. A cinética do Alox5 apresentou um aumento da expressão nos períodos de maior diferenciação celular em OBL WT. Além disso, a expressão de OCN, MMPs 2 e 9 e RANKL foram aumentadas em células 5-LO KO em quase todos os períodos avaliados. Em geral, o estímulo com LTs, seus inibidores e antagonistas diminuiu a expressão de Sp7, Col1a1, Opg e MMP-9 e aumentou RANKL em células KO. A sinalização por meio de segundos mensageiros também foi avaliada. Células 5-LO KO apresentam menor concentração de cálcio intracelular (Ca2+i) em relação ao WT. No período de 14 dias, o estímulo com LTD4 inibiu a liberação Ca2+i independente da linhagem, em relação ao controle. Os níveis de cAMP foram menores em OBL 5- LO KO, em todos os grupos tratados ou controle. LTD4 diminuiu a concentração de cAMP, mas não LTB4, em OBL 5-LO KO. O estudo também quantificou a produção de LTB4 e outros eicosanoides em osteoblastos mostrando a sua capacidade de síntese. A análise proteômica revelou 89 proteínas com expressão diminuída em OBL 5-LO KO, de um total de 154, sendo a maioria relacionada ao citoesqueleto e ao metabolismo energético. Também foram identificadas 59 proteínas exclusivas em OBL 5-LO KO e 06 unicamente expressas em células WT, revelando as diferenças intrínsecas de cada animal. O perfil osteoclastogênico de camundongos WT vs. 5-LO KO mostrou diferenças significativas na análise fenotípica, TRAP e na expressão gênica de células derivadas da linhagem monocítica-macrofágica. Após o estímulo com M-CSF e RANKL, as células WT apresentaram osteoclastos gigantes multinucleados, porém, células 5-LO KO apresentaram uma população de células com formas e tamanhos variáveis, e menor grau de maturação. Em adição, os LTsexógenos não modularam a atividade da TRAP. O meio condicionado proveniente dos OBL WT e KO, retardaram o processo de formação dos osteoclastos. A análise da expressão gênica em osteoclastos mostrou diminuição da expressão de Alox5, Il- 1b, Il-6 e TNFa em células 5-LO KO. BLT1/2, CysLt1 e os marcadores da diferenciação Acp5, Ctsk e Nfact1 não apresentaram diferenças entre os animais. Em adição, o LTB4 diminuiu a expressão do Alox5 e a Il-1b foi aumentada em osteoclastos WT. Assim, os resultados demonstram que os LTs são capazes de modular o metabolismo ósseo, e a ausência do gene da 5-LO está relacionada ao maior perfil osteogênico. / Leukotrienes (LTs) are inflammatory mediators derived from the 5-lipoxygenase (5-LO) pathway, with a relevant contribution in bone resorption. In this study we investigated the role of LTs in osteogenic differentiation and its impact on osteoclastogenesis.Thus, the bone profile of the 129/Sv (WT) and 5-LO Knockout mice (5-LO KO) was evaluated by computerized microtomography, showing higher vertebral bone density and thicker trabeculae in 5-LO KO males. After that, primary osteoblasts (OBL) were isolated and cultured to determine alkaline phosphatase activity (ALP) and mineralization potential. Results showed that OBL KO has higher ALP activity and mineralization, in all periods when compared with WT. In addition, the treatment with LTB4 and LTD4 inhibited calcium deposition. Inhibitors of LT synthesis and BLT1/2 antagonists were effective to recover the mineralized nodules formation. The kinetics of Alox5 showed an increase in expression during cellular differentiation period in WT OBL. In addition, expression of OCN, MMPs 2 and 9 and RANKL were increased in 5- LO KO cells in almost all evaluated periods. In general, the stimulation with LTs, their inhibitors and antagonists decreased the expression of Sp7, Col1a1, Opg and MMP- 9. But it increased the RANKL expression in KO cells. The second messengers signaling was also evaluated. 5-LO KO cells showed lower concentration levels of intracellular calcium (Ca2+ i) when compared to WT cells. In the 14-day period, the LTD4 treatment inhibited the Ca2+i independent of the murine lineage, relative to the control. cAMP levels were lower in OBL 5-LO KO, in all treated or control groups. LTD4 decreased the concentration of cAMP, but not LTB4, in KO cells. The study also quantified the production of LTB4 and other eicosanoids in osteoblasts showing their ability to synthesize those metabolites. The proteomic analysis revealed 89 downregulated proteins in OBL KO, out of a total of 154, most of them related to cytoskeleton and energy metabolism. Also 59 identified proteins were unique in OBL 5-LO KO and 06 exclusively expressed in WT cells, revealing the intrinsic differences of each strain. The osteoclastogenic profile of WT vs. 5-LO KO showed significant differences in phenotypic analysis, TRAP and in the gene expression of cells derived from the monocyte-macrophage-lineage. After M-CSF and RANKL stimulation, WT cells showed multinucleated giant osteoclasts. However, 5-LO KO cells presented a population of cells with variable shapes and sizes, and a lower maturation stage. In addition, exogenous LTs did not modulate TRAP activity. The conditioned medium from OBL WT and 5-LO KO delayed the formation process of osteoclasts. Gene expression analysis in osteoclasts showed decreased expression of Alox 5, Il-1b, Il-6 and TNF in 5-LO KO cells. BLT1/2, CysLt1 and the osteoclast differentiation markers Acp5, Ctsk and Nfact1 showed no differences between the strains. In addition, LTB4 decreased the expression of Alox5, and IL-1b was increased in WT osteoclasts. Thus, the results demonstrate that the LTs are able to modulate the bone metabolism, and the absence of the 5-LO gene is related to the greater osteogenic profile.

MMA (mixed martial arts) has expanded globally as entertainment, training and competition in recent years. MMA is a sport where techniques such as punches, kicks, throws and joints are used to defeat the opponent in unarmed close combat. Standing techniques such as punches and kicks are of great importance in sports and coaches / athletes strive to develop efficient techniques. The study investigates how UFC athletes work with distance moves relative to each other in knockout or tech knockout matches, comparisons were also made between a heavier and an easier weight class to detect any differences in behavior. Data was collected through a match observation where a total of 50 matches from the MMA event were collected with reverse chronological order starting from fight night Johnson VS Reis. The results showed that there were differences between movement and non-occupation at KO / TKO, but also that they were not significant. The same conclusion was made for comparisons between the weight classes. When there was an individual move, it was most common for it to move forward and the most common movement between athletes was forwards. Handling of dynamic and static distances is likely to have impact on elite athletes within MMA, but the results show that the difference between athletes in different weight classes is not significant and is likely to be affected to an equal extent by individual athletes' skills. / Kampsporten MMA (mixed martial arts) har expanderat globalt som underhållning, träning och tävling under de senaste åren. MMA är en sport där tekniker som slag, sparkar, kast och ledlås används för att besegra motståndaren i obeväpnad närkamp. Stående tekniker som slag och spark är av stor vikt inom sporten och tränare/atleter strävar efter att utveckla dessa tekniker på effektivaste sätt. Studien undersöker hur atleter inom UFC arbetar med distanshantering i förhållande till varandra vid matcher som avgjordes med knockout eller tekniskknockout, jämförelser gjordes också mellan en tyngre och en lättare viktklass för att påvisa eventuella skillnader i beteende. Data samlades in genom en matchobservation där totalt 50 matcher från MMA event samlades in med omvänd kronologisk ordning med start från galan fight night Johnson VS Reis. Resultatet visade att det fanns skillnader mellan rörelse och ickerörelse vid KO/TKO, men också att dessa inte var signifikanta. Samma slutsats gällde vid jämförelser mellan viktklasserna. När det skedde en enskild rörelse var det vanligaste att den skedde framåt och den vanligaste rörelsekombinationen mellan atleterna var framåt-framåt. Hantering av dynamisk och statiskdistans har med stor sannolikhet en betydande inverkan för elitatleter inom MMA, men resultatet visar att skillnaden mellan atleter i olika viktklasser inte är signifikant och påverkas troligen i lika stor utsträckning av individuella färdigheter hos atleterna.

L'apparition d'une protéinurie néphrotique constitue un effet secondaire aux thérapies ciblées anti-angiogéniques utilisées en oncologie (anti-VEGF ligand et les inhibiteurs de récepteurs tyrosine kinase). Dans ces travaux, nous avons étudié 29 patients traités par ces thérapies. 8 d'entre eux ont développé des lésions glomérulaires minimes ou hyalinose segmentaire et focale (LGM/HSF) majoritairement suite au traitement par les inhibiteurs des récepteurs tyrosine kinases (RTKIs) et 13 présentaient principalement des lésions de microangiopathie thrombotique (MAT) après thérapie anti-VEGF ligand. Dans cette étude, nous avons mis en évidence que c-mip est un acteur majeur du développement de protéinurie néphrotique consécutive aux traitements par les RTKIs. En effet, le Sorafenib (RTKI) induit l’expression de c-mip en inhibant l’activité de RelA et ce mécanisme serait impliqué dans le déclenchement des lésions LGM/HSF. Le rôle de c-mip dans la physiopathologie des podocytopathies acquises nous a conduit à générer un modèle murin d'invalidation de c-mip, conditionnelle et spécifique du podocyte. Les souris déficientes pour c-mip ont été étudiées dans deux modèles expérimentaux de protéinurie, induits par le LPS et le Sorafenib, respectivement. Dans les deux modèles, la protéinurie était significativement atténuée chez les souris déficientes avec préservation de l'architecture glomérulaire en comparaison des souris témoins. Le sorafenib a entraîné chez les souris témoins des lésions glomérulaires caractérisées par des rétractions du floculus, des thrombi intraglomérulaires et des lésions podocytaires. Ces résultats suggèrent que le sorafenib constitue un nouveau modèle murin d'induction d'une glomérulopathie expérimentale et que l'invalidation de c-mip spécifiquement dans le podocyte confèrerait une résistance au développement de protéinurie et de lésions rénales, suggérant que c-mip serait une cible thérapeutique potentielle. / Nephrotic proteinuria constitutes a serious side effect of anti-angiogenic therapies commonly used in oncology (anti-VEGF and tyrosine kinase receptorinhibitors, RTKI). In this work, we studied 29 patients treated by anti-angiogenic therapies. Eight of them developed minimal change nephrotic syndrome and focal and segmental glomerulosclerosis (MCNS/FSGS), mainly after RTKI treatment, and 13 underwent thrombotic microangiopathy lesions, mostly associated with anti-VEGF ligand therapy. C-mip overexpression was strongly related to the onset of nephrotic proteinuria after RTKI. Sorafenib (RTKI) induced c-mip expression by inhibiting RelA activity, ultimately leading to MCNS/FSGS. To confirm and clarify the pathophysiological role of c-mip in acquired podocytopathies, we generated a conditional, podocyte-specific c-mip knock-out mouse model. C-mip knockout mice were subjected to two experimental models of proteinuria, induced by LPS and Sorafenib, respectively. In each model, proteinuria was significantly decreased in cmip-invalidated mice, while glomerular architecture was preserved as compared to control mice. In the latter, Sorafenib led to glomerular tuft retractions, intraglomerular thrombi and podocyte lesions. This is suggested as the first experimental model of RTKI-induced glomerulopathy. Moreover, the podocyte specific knock out of c-mip confers resistance to proteinuria and renal injury, confirming c-mip as a potential therapeutic target in idiopathic nephrotic syndrome.

Les dommages de l'ADN peuvent s’accumuler dans les cellules souches hématopoïétiques(CSH) suite aux stress externes ou métaboliques et perturber leur fonctionnement et/ou leur maintien.La réparation par excision de nucléotides (NER), initiée par l’arrêt de la transcription (TCR) ou par lareconnaissance de distorsions des régions non transcrites (GGR) de l’ADN, est nécessaire àl’hématopoïèse à long terme. XPC, un facteur clé du système GGR, participe à d’autres réponses austress oxydatif. Le laboratoire a montré que la perte de XPC provoque l’accumulation de mutations, unstress métabolique et la carcinogenèse. Notre objectif est d’évaluer son expression et son rôle dans lemaintien et la différenciation des CSH. Nos résultats montrent qu’il est plus exprimé dans les cellulesimmatures CD34+ que dans les CD34- matures. Aussi, XPC apparaît sous trois poids moléculairesdifférents certainement liés à des modifications post-traductionnelles. Son extinction par ARNinterférence n'affecte ni la prolifération ni la capacité progénitrice in vitro des cellules CD34+.Cependant, les cellules déficientes implantées chez des souris immunodéficientes disparaissentprogressivement suggérant une perte des CSH ou de leur capacité de différenciation. Postulant queles mutations s’accumulent avec le temps, nous avons étudié l’hématopoïèse chez des sourisdéficientes en XPC jeunes et âgées. Les différences décrites dans l’hématopoïèse chez les individusjeunes et âgés sont retrouvées mais, de manière surprenante, aucune différence entre les animauxsauvages et mutés quelque soit l’âge ou le stress génotoxique n’est observée. Les résultats obtenussur les cellules humaines démontrent un rôle potentiel de XPC dans l’hématopoïèse, mais denouvelles investigations sont nécessaires pour mieux comprendre les mécanismes impliqués, et lapossible participation de XPC dans la leucémogenèse. / DNA damage may accumulate in hematopoietic stem cells (HSC) due to external ormetabolic stresses, leading to perturbation in their function and/or maintenance. Nucleotide excisionrepair (NER), initiated in the DNA by the stop of transcription (TCR) or by the recognition of distortionsin transcribed regions (GGR), is necessary for long-term hematopoiesis. XPC, a key factor in GGR, isimplicated in oxidative stress. The laboratory has demonstrated that XPC loss leads to theaccumulation of mutations, metabolic stress and carcinogenesis. Our objective is to evaluate XPCexpression and its role in HSC maintenance and differentiation. Results showed that XPC is highlyexpressed in immature CD34+ cells compared to mature CD34- cells. In addition, XPC appeared withthree different molecular weights, certainly linked to post-translational modifications. XPC silencing byshRNA did not affect the proliferation or the progenitor ability of CD34+ cells in vitro. However, deficientcells transplanted in immunodeficient mice disappeared progressively, suggesting the loss of HSCs ortheir differentiation capacity. Postulating that mutations accumulate with time, we have studiedhematopoiesis in young and aged XPC deficient mice. Differences described in young and agedhematopoiesis systems were found but, surprisingly, no difference was observed between wild typeand mutant mice at any age or genotoxic stress. Data from human cells demonstrate a potential rolefor XPC in HSC but new investigations are necessary to better understand the mechanisms implicatedand if XPC may participate in leukemogenesis.

Objective knowledge about quantity, composition and physicochemical characteristics of municipal solid waste (MSW) in the Czech Republic will form the basis for decision- making when considering processing facilities or regional waste management systems. As the country continues to streamline its legislation to the European Union's solid waste mandates, the results of these studies were employed by the Czech Ministry of Environment to optimise the national waste management strategy. This doctoral thesis focuses on the composition of MSW, primarily the mixed MSW from three types of households in the Czech Republic (urban, mixed and rural) differentiated by their heating methods and possibility of waste disposal. The respective experimental work and data-collection took place in years 2008 and 2009. Methodology used in the analyses is based on the sieve analysis of mixed MSW within predetermined grain size fraction and on the final manual sorting of the waste by material categories. The physicochemical characteristics were measured according to the standard analysis methods. The composition of mixed MSW in urban and mixed households are similar. Some of the biggest differences were found in the quantities of certain subsample categories, especially fine fraction (matter smaller than 8 mm), between...

<p>Cilj ovo rada je da obezbedi informacije o apstraktnim osobinama<br />klonova ko-operacija, mrezi klonova operacija, ali i ostalim objektima koji<br />prate pojam klona. U Uvodu je pokazano kako je nastao pojam ko-algebraske strukture. Kratko se razmatraju i dva pristupa ko-algebrama: Kleislijev, koji je potekao iz konteksta teorije kategorija [Kle 65], i Drbohlavov, koji je po osnovnoj ideji blizi<br />univerzalnoj algebri, i koji je usvojen u ovom radu. Glava 0 sadrzi spisak osnovnih pojmova i oznaka vezanih za skupove, preslikavanja, relacije, mreze i particije koji se koriste u celom radu, dok se u Glavama 1 i 2 uvode standardni termini teorije klonova operacija i ko-operacija i daje pregled poznatih rezultata. Originalni rezultati rada sadrzani su u Glavama 3&ndash;7. Glava 3 sadrzi osnovni rezultat rada. Pomocu kontravarijantnog liftinga kooperacija uspostavlja se izomorfizam izmedju klona svih ko-operacija skupa X s jedne strane i jednog posebnog klona operacija skupa P(X) sa druge strane. Pokazuje se da je isto preslikavanje ujedno i potapanje algebre kooperacija skupa X u algebru operacija skupa P(X) koja je standardna u teoriji klonova. Takodje se pokazuje da je kontravarijantni lifting klonova ujedno i izomorfizam mreze klonova ko-operacija skupa X i jednog glavnog ideala u mrezi klonova na P(X). Nakon ispitivanja lokalno ko-zatvorenih klonova ko-operacija pokazujemo u u kakvom odnosu stoje reprezentacije klonova ko-operacija selektivnim operacijama s jedne strane [Cs&acute;a 85], i operacijama na partitivnom skupu s druge strane. Reprezentaciju operacijama na partitivnom skupu, mada izomorfna jednom specijalnom slucaju reprezentacije selektivnim operacijama, smatramo bitnom, zato sto se njome klonovi ko-operacija smestaju u poznati ambijent skupovnih Booleovih algebri, umesto u prilicno opskuran prostor selektivnih operacija. U pretposlednjem odeljku se razmatra odnos klonova ko-operacija i klonova operacija kroz proces liftinga, sto je omoguceno cinjenicom da i kontravarijantni lifting klona svih ko-operacija i kovarijantni lifting klona svih operacija skupa X odredjuju klon operacija skupa P X. Na samom kraju ove pomalo dugacke glave se ispituju odnosi klonova ko-relacija i klonova relacija kroz prizmu monoida<br />transformacija. U Glavi 4 se izlazu neke osobine mreze klonova ko-operacija kao parcijalno uredjenog skupa. Opisani su intervali Int(M) za neke posebne monoide transformacija M. Pokazano je da u slucaju M = TX u mrezi klonova ko-operacijane postoji &ldquo;Burlova anomalija&rdquo; (ispitivanja kolapsirajucih monoida su, medjutim, odlozena do Glave 5). Nakon toga je predlozena jedna konstrukcija skupa nezavisnih ko-operacija na osnovu koje je dobijena donja granica za broj klonova ko-operacija na konacnom skupu i tacan broj klonova ko-operacija na beskonacnom skupu. Iako je dobijena donja granica prilicno neprecizna, na osnovu nje se sasvim jasno uocava &ldquo;veoma eksponencijalna priroda&rdquo; ovog broja. Ispitivanja jednog posebnog glavnog filtera mreze klonova ko-operacija nam daju gornju granicu za visinu mreze. Glava 5 je posve&acute;cena ispitivanjima maksimalnih klonova ko-operacija na kona-cnom skupu. Maksimalni klonovi ko-operacija su opisani u radu [Sz&acute;ek 89] kao skupovi operacija koji slabo cuvaju regularne familije skupova. Prvo se daje interpretacija ovog rezultata u terminima ko-relacija i pokazuje se da je ovakav opis najbolji moguci kada se u obzir uzme arnost dobijenih ko-relacija. Nakon toga se pokazuje da nijedan maksimalan klon nema Shefferovu ko-operaciju i daje se opis U&rdquo;&iexcl;1-maksimalnih klonova ko-operacija. Paznja se dalje prenosi na preseke maksimalnih klonova ko-operacija. Prvo se razmatraju preseci nekih parova maksimalnih klonova i pokazuje se da to ne mora uvek biti maksimalni klon, a kasnije se konstruiˇse potapanje mreze cLB1 &pound; : : : &pound; cLBk u mrezu cLX. Potom se pokazuje da je mreza cLX komplementirana, a na samom kraju se razmatraju kolapsirajuci klonovi ko-operacija i monoidi. Glava 6 je posvecena opisu minimalnih klonova ko-operacija i svih supminimalnih klonova ko-operacija koji nisu esencijalno unarni. Uvid u strukturu supminimalnih klonova omogucuje izvodjenje donje granice za visinu mreze klonova ko-operacija. Glava sadrzi i kratak komentar o asocijativnosti kooperacija. U Glavi 7 se razmatraju enumerativne osobine mreze klonova ko-operacija na troelementnom skupu, kao i nekih njenih pratecih objekata. Osim utvrdjivanja kardinalnosti mreze na skupu f0; 1; 2g, odredjena je njena visina, kao<br />i svi submaksimalni klonovi. Poseban odeljak je posvecen enumeraciji baz&macr;a<br />klona svih ko-operacija i svih maksimalnih klonova, klasicnoj temi u teoriji<br />klonova. U dodacima su navedene tabele kojima se sumarizuju rezultati ove<br />glave i dat je opis jednostavnog softverskog alata koji ima ulogu &ldquo;raˇcunarskog<br />atlasa&rdquo; mreˇze cL3. S obzirom da mreza ima previse elemenata da bi se mogao<br />dati njen Hasse dijagram, &ldquo;racunarski atlas&rdquo; se pokazao kao najjednostavniji<br />i najefikasniji nacin da se dodje do informacija o mrezi. Napomenimo da su mnogi rezultati u ovom radu dobijeni uopstavanjem osobina mreze koje su otkrivene &ldquo;prelistavanjem atlasa&rdquo;. Navedimo kao primer strukturu U&rdquo;&iexcl;1- maksimalnih klonova, gornju i donju granicu za visinu mreze, cinjenicu da je mreza komplementirana, kao i to da maksimalni klonovi nemaju Shefferovu ko-operaciju.</p> / <p>The aim of this thesis is to provide information on abstract properties of clones of co- operations, the lattice of clones of co-operations and other accompanying objects.&nbsp; Introduction to the thesis demonstrates in short the genesis of concepts of co-operation and co-algebra and presents two approaches to the topic: the Kleisli&nbsp; approach which originated in the category theory, and the approach<br />of Drbohlav which is more in the spirit of universal algebra and which is adopted in the thesis.<br />Chapter 0 is a short display of standard set-theoretic terminology and notation which is used in the thesis.<br />Chapters 1 and 2 present standard notions of theory of clones of operations and co-operations, respectively. They contain lists of the most important known results. The original contribution of this thesis is contained in Chapters 3&ndash;7.<br />Chapter 3 contains the basic result of the thesis. By means of contravariant lifting of co-operations we establish an isomorphism between the clone&nbsp; of all co-operations on a set X and one special clone of operations on the set P(X). This isomorphism is not only the abstract clone isomorphism, but also a lattice isomorphism between the lattice of all clones of co-operations on X and a principal ideal in the lattice of all clones of operations on P(X). The same mapping is an embedding of the algebra of co-operations on X into the algebra of operations on P(X). Locally co-closed clones of operations are also characterised via this most useful mapping. Representation of clones of co-operations by operations on the powerset is compared to the representation by selective operations. Although isomorphic to a special case of the representation by selective operations, representation by operations on the powerset is highly important because it places clones of co-operations into a familiar setting of set-theoretic Boolean algebras, rather then in the quite obscure setting of selective algebras. At the end of this lengthy chapter, we investigate the relationship of corresponding liftings of the clone of all operations and the clone of all co-operations, and ellaborate the lifting proces and the interplay between description of transformation monoids by relations and co-relations.<br />Chapter 4 exibits some order-theoretic properties of the lattice of clones of co-operations. Intervals of the form Int(M) are described for some special transformation monoids M. In case M = TX it is demonstrated that the socalled &ldquo;Burle anomaly&rdquo; does not occur in the lattice of clones of co-operations. The investigation of collapsing clones and monoids is, however, deferred until&nbsp; Chapter 5. After that a construction of an independent set of co-operations is presented, based on which a lower bound for the number of clones of cooperations is obtained. Although pretty rough, this lower bound shows that the number of clones of co-operations on a finite set is of a &ldquo;very exponental nature&rdquo;. The number of clones of operations on an infinite set is also obtained. The investigations of a particular principal filter of the lattice provide an upper bound for the height of the lattice. Chapter 5 is devoted to the investigation of maximal clones of co-operations on a finite set. All the maximal clones of co-operations are described in [Sz&acute;ek 89] in terms of regular families. We first reinterpret that result in terms of co-relations and show that the description is the best possible as far as arities of co-relations involved are considered. After that we supply some more information on the maximal clones. We show that no maximal clone of co-operations has a Sheffer co-operation and describe clones covered by U&rdquo;&iexcl;1.<br />Then we turn to intersections of maximal clones of co-operations. First we consider intersections of some special pairs of maximal clones and show that in some cases this is not a maximal clone. As for the intersection of several maximal clones of co-operations, we show how to embed cLB1 &pound; : : : &pound; cLBk<br />in cLX. Using results on the structure of maximal clones we show that the lattice cLX is complemented. The chaptr ends with a note on collapsing clones of co-operations.<br />Chapter 6 provides the description of minimal clones of co-operations and those supminimal clones of co-operations which are not essentially unary. The structure of supminimal clones of co-operations provides a lower bound for the height of the lattice of clones of co-operations. As a spin-off, there is<br />a brief discussion on associativity.<br />Chapter 7 is devoted to enumerations of various objects connected to the lattice of clones of co-operations on a three element set. Besides the enumeration of the lattice itself, the submaximal clones have been listed and the height of the lattice on a three element set is determined. A separate section deals with a classical clone-theoretic topic of enumerating the bases for the clone of all co-operations and for the maximal clones of co-operations. The Appendices to this chapter contain some tables that summarize various enumerations, as well as a description of a modest software tool that palys the role of the &ldquo;computer atlas&rdquo; of cL3. Since the lattice has too many elements to be drawn explicitely, this was the simplest and the most efficient way to handle it. Let us remark that many results of this thesis were obtained by gathering the information on the particular case from the &ldquo;computer atlas&rdquo;<br />and by generalisation, such as: the structure of U&rdquo;&iexcl;1 maximal clones, the bounds for the height of the lattice, the fact that maximal clones have noSheffer co-operation and the fact that cLX is complemented.</p>

Based on accumulating evidence, adequate intake of whole grains is associated with reduced cardiovascular disease CVD risk. Germinated brown rice (GBR) has been used in East Asian countries as an alternative grain. Preliminary studies suggest GBR has potential health benefits, including reducing CVD risk, but the mechanism remains unclear. The hypothesis of the project is that long-term consumption of GBR would reduce atherogenic risk factors in low-density lipoprotein receptor knockout (LDLr-KO) mice. To test the hypothesis, three groups of male LDLr-KO mice were fed with one of the following diets for 24 weeks: (a) commercial mouse chow, used as the control diet; (b) chow was replaced with 60% (w/w) Chinese white rice (CWR); and (c) chow was replaced with 60% (w/w) GBR. All diets were supplemented with 0.06% (w/w) dietary cholesterol to accelerate atherogenesis. Blood samples, hearts, livers and feces were collected and used for biochemical and histological analyses. The results demonstrated that no significant difference was detected in body weights, plasma or fecal lipid profiles and antioxidant enzyme activities among groups. However, GBR consumption significantly decreased atherosclerotic lesion (P = 0.003) in the aortic roots as compared with that in the CWR group, but there was no significant difference as compared with that in the control group (P = 0.4). In addition, GBR significantly decreased monocyte adhesion to the aorta in LDLr-KO mice as compared to that in the CWR group (P=0.0001), but not with the control group. These data suggested that GBR may be beneficial for the prevention of vascular inflammation and atherogenesis in LDLr-KO mice. Additional studies in animal models and humans may further investigate the mechanisms of the beneficial effects of GBR on vascular inflammation and atherogenesis. / May 2017