Submitted by JÚLIO HEBER SILVA (julioheber@yahoo.com.br) on 2016-12-06T16:54:56Z No. of bitstreams: 2 Dissertação - Bruno César Ferreira Gonzaga - 2016.pdf: 2851863 bytes, checksum: 3316d056c52b9a8ca6ed7608909e2b80 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Jaqueline Silva (jtas29@gmail.com) on 2016-12-13T15:35:12Z (GMT) No. of bitstreams: 2 Dissertação - Bruno César Ferreira Gonzaga - 2016.pdf: 2851863 bytes, checksum: 3316d056c52b9a8ca6ed7608909e2b80 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-12-13T15:35:12Z (GMT). No. of bitstreams: 2 Dissertação - Bruno César Ferreira Gonzaga - 2016.pdf: 2851863 bytes, checksum: 3316d056c52b9a8ca6ed7608909e2b80 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-09-30 / Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG / The Brachiaria grass, with broad geographic distribution in Brazil, has been shown to cause photosensitization and appearance of foamy macrophages in the liver and liver and mesenteric lymph nodes of ruminants. Weight loss of 2.52% in cattle with subclinical hepatogenous photosensitization was documented and it is estimated that in one year in a single slaughterhouse more than R$ 100,000 in annual losses are caused by liver damage resulting from consumption of Brachiaria spp. The aim of this study was to corelate the extent of liver parenchyma containing foamy macrophages with hot carcass weight of cattle. Carcasses of cattle aged 20-36 months were weighed and divided into two groups: Group 1 (544.87-607.13 pounds) and Group 2 (689.42-757.24 pounds) The livers of bovines were weighed and samples of the right lobe were collected for histopathologic analysis. The areas of hepatic parenchyma containing foamy macrophages were measured using the ImageJ® program. Statistical analysis of the parameters (liver weight, hot carcass weights and affected area of liver tissue) was performed quantitatively, by calculating the mean, standard deviation and coefficient of variation. These data were analyzed for normality and homogeneity of variances, through the Kolmogorov-Smirnov and Bartlett tests. As the variables were homogeneous and had a normal distribution were evaluated by parametric test analysis of variance. The correlation of variables was performed by Spearman test. The significance level was set at 5% and all descriptive analyzes were performed by EXCEL program and statistical tests by the software R. Cattle grazing Brachiaria spp. presented hepatic lesions characterized by degeneration, necrosis, presence of mild inflammatory infiltrates and small area of hepatic tissue containing foamy macrophages. There was no influence of the affected area of the hepatic parenchyma containing foamy macrophages with hot carcass weight of cattle. / O capim braquiária, com ampla distribuição geográfica no Brasil, vem sendo apontado como causa de fotossensibilização e do aparecimento de macrófagos espumosos no fígado e linfonodos hepáticos e mesentéricos de ruminantes. Foram relatadas perdas de peso na ordem de 2,52% em bovinos com fotossensibilização hepatógena subclínica e estimou-se que mais de R$ 100.000 em perdas anuais por lesões hepáticas em apenas um frigorífico fosse decorrente do consumo de Brachiaria spp. O objetivo deste estudo foi correlacionar a extensão do parênquima hepático contendo macrófagos espumosos com o peso de carcaça quente dos bovinos. As carcaças dos bovinos, com idade entre 20 a 36 meses, foram pesadas e distribuídas em dois grupos: Grupo 1 (247,15 a 275,39 kg) e Grupo 2 (312,72 a 343,48 kg). Os fígados dos bovinos foram pesados e amostras do lobo direito coletadas para análise histopatológica. As áreas do parênquima hepático contendo macrófagos espumosos hepáticos foram mensuradas utilizando-se o programa ImageJ®. A análise estatística dos parâmetros (peso do fígado, peso da carcaça quente e área comprometida do tecido hepático) foi realizada de forma quantitativa, calculando a média, desvio padrão e coeficiente de variação. Esses dados foram analisados quanto à normalidade e homogeneidade das variâncias, por meio dos testes de Kolmogorov-Smirnov e Bartlett. Como as variáveis foram homogêneas e tiveram uma distribuição normal, foram avaliadas pelo teste paramétrico de análise de variâncias. A correlação das variáveis foi realizada pela correlação de Spearman. O grau de significância adotado foi de 5% e todas as análises descritivas foram realizadas pelo programa EXCEL e os testes estatísticos pelo programa R. Os bovinos sob pastejo de Brachiaria spp. apresentaram lesões hepáticas caracterizadas por degeneração, necrose, presença de infiltrados inflamatórios de intensidade leve e pequena área do tecido hepático contendo macrófagos espumosos. Não houve influência da área comprometida do parênquima hepático contendo macrófagos espumosos com o peso de carcaça quente dos bovinos.

Submitted by ANA CAROLINA ORTEGAL ALMEIDA null (ortegalmedvet@gmail.com) on 2016-03-21T11:23:32Z No. of bitstreams: 1 Dissertação_Ana_Carolina_Ortegal_Almeida.pdf: 2529882 bytes, checksum: 7d592a7135492a17094d124933d025a2 (MD5) / Approved for entry into archive by Juliano Benedito Ferreira (julianoferreira@reitoria.unesp.br) on 2016-03-22T14:01:48Z (GMT) No. of bitstreams: 1 almeida_aco_me_jabo.pdf: 2529882 bytes, checksum: 7d592a7135492a17094d124933d025a2 (MD5) / Made available in DSpace on 2016-03-22T14:01:48Z (GMT). No. of bitstreams: 1 almeida_aco_me_jabo.pdf: 2529882 bytes, checksum: 7d592a7135492a17094d124933d025a2 (MD5) Previous issue date: 2016-01-20 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A inspeção da carcaça e órgãos nos abatedouros objetiva limitar o aproveitamento de produtos impróprios para consumo humano, protegendo a população contra doenças transmitidas pelos alimentos. O fígado é uma víscera nutritiva e bem aceita pelos consumidores, mas suas funções metabólicas o tornam susceptível a diversas lesões, fazendo com que seja condenado frequentemente na rotina de inspeção. Este estudo analisou fígados bovinos e relacionou as lesões macro e microscópicas com os sistemas de produção dos animais abatidos (sistemas intensivo, semi-intensivo e extensivo). Avaliou também a presença de lesão microscópica em fígados sem lesão macroscópica. Foram coletadas 450 amostras de fígado bovino dos diferentes sistemas de produção, as quais foram processadas para exame histológico. As alterações macroscópicas observadas nos fígados bovinos foram: fibrose capsular (32%), aderências/peri-hepatite (25,8%), teleangiectasia (24,4%), manchas pálidas ou amareladas (20,9%), hemorragia subcapsular (7,1%), abscessos (5,3%), granulomas (3,5%), congestão (1,8%) e cistos (0,9%). Os achados microscópicos foram: infiltrado inflamatório mononuclear (66,4%), macrófagos espumosos (40,9%), tumefação hepatocelular (30%), tumefação hepatocelular centrolobular (22,7%), fibrose capsular (26,4%), degeneração gordurosa (15,1%), peri-hepatite (13,8%), necrose (13,5%), teleangiectasia (12,2%), abscessos (2,9%), granulomas (1,8%), congestão (16%), hemorragia subcapsular (4,6%), infiltrado inflamatório polimorfonuclear (2,5%), infiltrado inflamatório misto (8,7%), hiperplasia ductal (4,9%) e cistos (0,9%). As lesões que apresentaram relação com os sistemas de produção (P<0,05) foram: infiltrado inflamatório mononuclear, macrófagos espumosos e tumefação hepatocelular. Os fígados de bovinos criados em sistema intensivo apresentaram mais lesões macroscópicas e os fígados de bovinos criados em sistema extensivo apresentaram mais lesões microscópicas. Somente 19,5% dos fígados sem lesão macroscópica não apresentaram nenhum tipo de lesão microscópica. Conclui-se que o sistema de produção pode influenciar na ocorrência de algumas lesões hepáticas e que fígados sem lesão macroscópica, liberados para consumo humano, podem apresentar várias lesões microscópicas importantes. / The inspection of cattle carcasses and organs at slaughterhouses aims at limiting the use of products with abnormalities that makes them improper for human consumption, thereby protecting the public from food-borne diseases. The liver is a nutritious viscera and it is well accepted by consumers. However, its metabolic functions make it susceptible to various injuries, which often condemn it during inspections routine. This study evaluated bovine livers and linked the macro and microscopic lesions found to the beef cattle production systems (intensive, semi intensive and extensive). Also evaluated the presence of microscopic lesions on livers without macroscopic findings. Some 450 samples of beef liver were collected from various production systems and were processed for histological examination. Macroscopic changes observed on the samples were: capsular fibrosis (32%), adhesions/perihepatitis (25.8%), telangiectasia (24.4%), pale or yellowish spots (20.9%), subcapsular hemorrhage (7.1%), abscesses (5.3%), granulomas (3.5%), congestion (1.8%) and cysts (0.9%). Microscopic findings were mononuclear cell infiltration (66.4%), foamy macrophages (40.9%), hepatocellular swelling (30%), centrilobular swelling (22.7%), capsular fibrosis (26.4%), fatty degeneration (15.1%), perihepatitis (13.8%), necrosis (13.5%), telangiectasia (12.2%), abscesses (2.9%), granulomas (1.8%) congestion (16%), subcapsular hemorrhage (4.6%), polymorphonuclear inflammatory infiltrate (2.5%), mixed inflammatory infiltrate (8.7%), ductal hyperplasia (4.9%) and cysts (0.9%). Those injuries related to production systems (P<0.05) were: mononuclear cell infiltration, foamy macrophages and cellular swelling. The livers of bovines raised in extensive system present more macroscopic lesions and the livers of bovines raised in intensive system present more microscopic lesions. Only 19.5% of livers without macroscopic findings did not show histopathological lesions. This study concludes that beef cattle production systems can influence the occurrence of some liver lesions and that liver without macroscopic findings, released for human consumption may have several important microscopic lesions.

L'une des particularités de Mycobacterium tuberculosis, agent pathogène de la tuberculose, est sa capacité à accumuler des lipides dans son cytoplasme, ce qui favorise son entrée en dormance. Le séquençage du génome de M. tuberculosis a permis d'identifier certains gènes codant pour des enzymes lipolytiques, parmi ceux-ci : le gène codant pour la protéine Rv3097c aussi appelée LipY (composée d'un domaine PE et d'un domaine lipase relié par un Linker). Dans la première partie de ce travail de thèse, nous avons procédé à la caractérisation biochimique de LipY, mais aussi de ses formes mutantes LipY(∆PE), LipY(∆149) et LipY(∆170), et à l'étude d'inhibition des membres de la famille Lip apparentés à la lipase hormono-sensible humaine (Lip-HSL). Nous avons pu déterminer les propriétés cinétiques de l'activité lipase de LipY et de ses différentes formes mutantes.Dans la seconde partie, nous nous sommes intéressés à la compréhension du rôle des différents domaines de LipY, et du linker dans l'hydrolyse des lipides au cours de l'infection en utilisant des macrophages spumeux (macrophage riche en lipides) infectés au préalable par des souches de M. bovis BCG recombinantes. Ces résultats et les hypothèses posées durant ce travail de thèse, pourraient être appuyés par l'obtention de la structure tridimensionnelle de LipY. Pour cela, nous avons initié et procédé à la cristallogenèse de LipY. La poursuite des études d'optimisation des cristaux obtenus pourrait permettre d'aller plus en profondeur dans l'élucidation du rôle et du mécanisme d'action de LipY. / Mycobacterium tuberculosis is a pathogenic agent, responsible of the tuberculosis, which can store lipids into the cytoplasm. This accumulation allows the bacteria to enter in the dormancy phase. The sequencing of M. tuberculosis genome, allows to identify some genes coding for lipolytic enzymes, among which a gene coding for Rv3097c protein, also called LipY. (possessing PE domain linkto a lipase domain). During my PhD thesis, we first biochemically characterized LipY and its mutant forms LipY(ΔPE); LipY(Δ149) and Lip(YΔ170) and studied the inhibition of Lip family members related to the human hormone-sensible lipase (Lip-HSL). We determined the kinetic properties for the lipase activity of LipY and its mutants. In the second part, based on these previous results, we studied the role of these different domains and the linker on the hydrolysis of lipids during the infection phase, in infected foamy macrophages (lipids rich). For these studies, we used foamy macrophages infected by recombinants strains of M. bovis BCG (LipY(wt) and its mutants.These results and hypothesis can be confirmed and supported by resolving the tridimensional structure of LipY. Crystallogenesis tests allowed us to have some crystals of LipY(wt), which after optimization would allow us to have a better understanding of the role and action mechanism of LipY.

<p dir="ltr">Though mammalian spinal cord injury (SCI) has long been a topic of study, effective therapies that promote functional recovery are not yet available. The axolotl, <i>Ambystoma mexicanum</i>, is a valuable animal model in the investigation of spinal cord regeneration, as this urodele is able to achieve functional recovery even after complete spinal cord transection. Understanding the similarities and differences between the mammalian SCI response and that of the axolotl provides insight into the process of successful regeneration, and bolsters the fundamental knowledge used in the development of future mammalian SCI treatments. This thesis provides a detailed analysis of the ultrastructure of the axolotl meninges, as this has not yet been presented in existing literature, and reveals that the axolotl meninges consist of 3 distinct layers as does mammalian meninges; the dura mater, arachnoid mater, and pia mater. The role of reactive meningeal and ependymal cells is also investigated in regard to the deposition and remodeling of the fibrotic ECM, which is found to be similar in composition to hydrogel scaffolds being studied in mammalian SCI. It is shown that meningeal fibroblasts are the primary source of the extensive fibrillar collagen deposition that fills the entire spinal canal, peaking at approximately 3 weeks post transection and remaining until approximately 5 weeks post transection, and that there is no deposition of type IV collagen within the lesion site. Mesenchymal ependymal cells are shown to contribute to the ECM deposition through the production of glycosaminoglycans that are used in sidechains of both unsulfated and sulfated proteoglycans, while simultaneously remodeling the ECM through the production of MMPs and phagocytosis of cellular debris. Further, this study shows that mesenchymal ependymal cells and a population of foamy macrophages contribute to the degradation of the fibrin clot that forms in the acute phase of injury, and that this fibrin clot provides a necessary and permissive substrate for early mesenchymal outgrowth.</p>