Submitted by Sara Ribeiro (sara.ribeiro@ucb.br) on 2017-09-04T12:26:15Z No. of bitstreams: 1 CelsoAntonioPiresFerreiraDissertacao2016.pdf: 1073723 bytes, checksum: ce1f93231c8de5e5a55f8b87e4d88d7b (MD5) / Approved for entry into archive by Sara Ribeiro (sara.ribeiro@ucb.br) on 2017-09-04T12:26:24Z (GMT) No. of bitstreams: 1 CelsoAntonioPiresFerreiraDissertacao2016.pdf: 1073723 bytes, checksum: ce1f93231c8de5e5a55f8b87e4d88d7b (MD5) / Made available in DSpace on 2017-09-04T12:26:24Z (GMT). No. of bitstreams: 1 CelsoAntonioPiresFerreiraDissertacao2016.pdf: 1073723 bytes, checksum: ce1f93231c8de5e5a55f8b87e4d88d7b (MD5) Previous issue date: 2016-06-03 / The regional integration system that is based on the European Union was the result of a strong mutation in classical conceptual framework of the Nation. Sovereignty, according to the Westphalian???s concept began to be relaxed because of the internationalization of supranational rules based on cooperation between governments. In the European bloc, because of the economic crisis, it was found that the centralized political system in supranational entity began to be faced with structural problems of legitimacy, to be exercised by the States. These problems were consequences of the attempt of an economic and social planning within the European bloc. Experiment in which a reduction of economic imbalances have been made through paternalism, throughout the States economically stronger provide financial assistance during the integration process, the economically weaker countries. Such actions of social policy have proven insufficient to maintain economic stabilization block when member states are inserted in a recessionary environment. It was found that now, the States economically stronger tend to influence the decisions of the European block and remove the self-management capacity of the economically weaker states, leading to submission of central supranational decision models. In this process, a mutation in the interpretative content of the historical concept of sovereignty is unfolding. The difficulty of weak states in overcoming the economic crisis and internalize Community law produced by these supranational organisms has led to an ideology of resurgence of nationalist discourse. Authoritative this juncture, the danger of fragmentation of European unity can become a growing possibility. / O sistema de integra????o regional, em que se baseia a Uni??o Europeia, foi resultado de uma forte muta????o na estrutura conceitual cl??ssica do Estado na????o. A soberania, segundo o conceito westfaliano, come??ou a ser flexibilizada em virtude da internacionaliza????o de normas supranacionais baseadas na coopera????o entre governos. No bloco europeu, em virtude das crises econ??micas, verificou-se que o sistema pol??tico centralizado na entidade supranacional come??ou a se deparar com dificuldades estruturais de legitima????o, a serem exercidas pelos Estados membros. Esses problemas foram consequ??ncias da tentativa de uma planifica????o econ??mico-social dentro do bloco europeu. Experimento no qual a diminui????o dos desequil??brios econ??micos tem sido feita por meio de assistencialismos, por meio dos quais os Estados membros economicamente mais fortes providenciam ajuda financeira durante o processo de integra????o, aos pa??ses economicamente mais fracos. Tais a????es de pol??tica social t??m se mostrado insuficientes para manter a estabiliza????o econ??mica do bloco quando os Estados membros se encontram inseridos em um contexto recessivo. Verificou-se que, nesta conjuntura, os Estados economicamente mais fortes tendem a influenciar as decis??es do bloco europeu e retiram a capacidade de autogest??o dos Estados economicamente mais fracos, conduzindo-os ?? submiss??o dos modelos de decis??o supranacional central. Nesse processo, uma muta????o no conte??do interpretativo do conceito hist??rico de soberania vem se revelando. A dificuldade dos Estados fracos em superar as crises econ??micas e internalizar o direito comunit??rio produzido pelos ??rg??os supranacionais tem conduzido estes a uma ideologia de recrudescimento do discurso nacionalista. Abalizado nesta conjuntura, o perigo da fragmenta????o da unidade europeia pode se tornar uma possibilidade crescente.

Submitted by Sara Ribeiro (sara.ribeiro@ucb.br) on 2017-09-08T18:11:30Z No. of bitstreams: 1 OrzenilBonfimdaSilvaJuniorTeseParcial2017.pdf: 781918 bytes, checksum: 8eef627ca550957f6bfa1f46e54c687c (MD5) / Approved for entry into archive by Sara Ribeiro (sara.ribeiro@ucb.br) on 2017-09-08T18:11:38Z (GMT) No. of bitstreams: 1 OrzenilBonfimdaSilvaJuniorTeseParcial2017.pdf: 781918 bytes, checksum: 8eef627ca550957f6bfa1f46e54c687c (MD5) / Made available in DSpace on 2017-09-08T18:11:38Z (GMT). No. of bitstreams: 1 OrzenilBonfimdaSilvaJuniorTeseParcial2017.pdf: 781918 bytes, checksum: 8eef627ca550957f6bfa1f46e54c687c (MD5) Previous issue date: 2017-07-11 / In the last twenty-five years, we have witnessed the wide adoption of DNA markers for the study of genetic variation in many organisms. A DNA marker must have two or more identifiable allelic DNA sequences to be useful. It usually does not have a biological effect, but instead functions as a traceable landmark in the genome, found in a specific location, and transmitted by the standard laws of inheritance from one generation to the next. Its application goes beyond genetic mapping and includes the analysis of genetic diversity, marker-trait association studies, marker assisted selection and, more recently, with the advent of wholegenome sequencing, whole-genome association and genomic selection. Among the several types of DNA sequence polymorphisms that can be used as DNA marker, Single Nucleotide Polymorphisms (SNPs) are the most powerful for large-scale variation analysis. There are vast numbers of SNPs in every genome and they can be typed by methods that have been proven easy to automate. Detection of alternative alleles is rapid and effortless because it is based on well-known polymerase chain reaction and DNA oligomer hybridization assays. Various strategies have been devised to discriminate alleles at a SNP, including fixed DNA arrays technologies, solution hybridization techniques and many sequencing-based genotyping. In our study, we have developed high-throughput DNA marker systems for non-model, highly heterozygous, diploid tree species. We took advantage of the combined power of Next Generation Sequencing (NGS) technologies, well-established highly automated methods of SNP typing and bioinformatics algorithms to perform genome-wide DNA variation analysis. We used whole genome resequencing of pooled individuals to develop a high-density 60K SNP chip for Eucalyptus species (EucHIP60k) providing a 96% genome-wide coverage with 1 SNP/12???20 kbp, and 47,069 SNPs at ??? 10 kb from 30,444 of the 33,917 genes in the Eucalyptus genome. We then used high-density SNP data and whole-genome pooled resequencing to examine the landscape of population recombination (??) and theta (??), assess the extent of linkage disequilibrium (r2) and build the highest density linkage maps for Eucalyptus to date. Chromosome-wide ancestral recombination graphs allowed us to date the split of Eucalytpus grandis (1.7???4.8 million yr. ago) and identify a scenario for the recent demographic history of the species. In a final set of studies, we built the first genome assembly for a Neotropical forest tree, the Pink Ip?? (Handroanthus impetiginosus), a highly-valued keystone timber species. Genome sequence was screened for the development of a targeted-capture sequencing system for SNP genotyping consisting of nearly 24,000 probe sequences. This genotyping system showed flexibility as it allowed the identification of SNPs across different populations of the species in moderate sample sizes. The good genome coverage, consistent Ts/Tv ratio estimated across samples and fair technical reproducibility between replicates, in terms of recall and precision of the SNP calling and accuracy on genotypes, indicate that this genotyping platform can be confidently used to estimate population genetics parameters and carry out population genomics investigations at the genome-wide scale / ***

Submitted by Sara Ribeiro (sara.ribeiro@ucb.br) on 2018-02-16T15:39:34Z No. of bitstreams: 1 GilceliadePaulaSantosDissertacaoParcial2017.pdf: 1342828 bytes, checksum: 280d86560833a80e06f4a639d91b59e9 (MD5) / Approved for entry into archive by Sara Ribeiro (sara.ribeiro@ucb.br) on 2018-02-16T15:39:42Z (GMT) No. of bitstreams: 1 GilceliadePaulaSantosDissertacaoParcial2017.pdf: 1342828 bytes, checksum: 280d86560833a80e06f4a639d91b59e9 (MD5) / Made available in DSpace on 2018-02-16T15:39:42Z (GMT). No. of bitstreams: 1 GilceliadePaulaSantosDissertacaoParcial2017.pdf: 1342828 bytes, checksum: 280d86560833a80e06f4a639d91b59e9 (MD5) Previous issue date: 2017-11-29 / The present work consists of two essays and an experiment in which the behavior of the agents of the real estate market (Financial Institutions and alleged borrowers) is analyzed, using the Economic Analysis of Law combined with the Analytical-Behavioral Theory of Law. The objective of the work is to identify how these agents behave through the incentives granted by the Minha Casa, Minha Vida government program and the consequences of these behaviors in the real estate market. These studies are justified since the Federal Government has been working with the objective of reducing the country's housing deficit by means of actions to subsidize the acquisition of housing units for the lower income population, which are guided by a vast set of legal, Guiding the performance of individuals, builders and financial institutions, which ended up influencing the market, understood here as free interaction among the agents to perform barter trade. / O presente trabalho comp??e-se de dois estudos e um experimento, nos quais se analisa o comportamento dos agentes do mercado imobili??rio, institui????es financeiras e pretensos mutu??rios, utilizando-se a An??lise Econ??mica do Direito ??? AED. O objetivo dos trabalhos ?? identificar como comportam-se esses agentes mediante os incentivos concedidos pelo programa de governo Minha Casa Minha Vida e as consequ??ncias desses comportamentos no mercado imobili??rio. Estes estudos se justificam uma vez que o Governo Federal vem atuando com objetivo de reduzir o d??ficit habitacional do pa??s, por meio de a????es de subs??dios ?? aquisi????o de unidades habitacionais ?? popula????o de menor renda. Estas a????es s??o norteadas por um vasto conjunto de regras jur??dicas, guiando a atua????o de indiv??duos, construtoras e institui????es financeiras, que acabam por influenciar o mercado, aqui compreendido como intera????o livre entre os agentes para realiza????o de trocas por meio de barganha.

Made available in DSpace on 2015-04-14T13:34:31Z (GMT). No. of bitstreams: 1 403013.pdf: 1643144 bytes, checksum: 5ccd3fb28b7fe3934f949e3f92a8d4ac (MD5) Previous issue date: 2008-03-28 / A fam?lia erbB ? composta por quatro membros: erbB1, erbB2, erbB3 e erbB4. Essa fam?lia tem aparecido amplamente em diversos trabalhos, por?m, o estudo particular da prote?na erbB3 no c?ncer de pulm?o ainda permanece muito restrito. O objetivo desse estudo foi descrever a freq??ncia de express?o da prote?na erbB3 em amostras de c?ncer de pulm?o n?o-pequenas (NSCLC) c?lulas, assim como correlacionar os achados da freq??ncia de express?o da prote?na com as seguintes caracter?sticas clinico-patol?gicas dos pacientes como: idade, g?nero, tipo histol?gico, estadiamento patol?gico, grau histol?gico, ?bito e sobrevida. Um total de 66 esp?cimes foram analisados por imuno-histoqu?mica. A freq??ncia de express?o da prote?na erbB3 esteve presente em 57 casos (86,4%). N?o foi encontrada rela??o estatisticamente significativa entre a freq??ncia de express?o da prote?na e as vari?veis: idade, g?nero, tipo histol?gico, estadiamento, grau histol?gico, ?bito e sobrevida. Entretanto, quando as amostras foram agrupadas em categorias de express?o entre 0 e 75% e 75 e 100% das c?lulas tumorais positivas para a prote?na erbB3, houve signific?ncia estat?stica com rela??o ao grau histol?gico (P=0,04) sendo o de maior freq??ncia o grau histol?gico tipo II (72,5%). Embora n?o tenha sido poss?vel correlacionar a presen?a da express?o da prote?na erbB3 com o progn?stico e os outros par?metros cl?nicos, provavelmente devido ao pequeno n?mero de amostras investigadas neste estudo, foi poss?vel observar a presen?a desta prote?na em 86,4% das amostras estudadas, comprovando que, assim como o EGFR e o erbB2, o erbB3 pode ser encontrado no NSCLC de forma bastante pronunciada. ? poss?vel que informa??es relativas ? express?o dos outros membros desta fam?lia nestas mesmas amostras, possam trazer novas interpreta??es, lembrando, novamente, que seus membros, j? caracterizados como prote?nas participantes na oncog?nese, atuam de forma sin?rgica

Made available in DSpace on 2015-04-14T13:34:44Z (GMT). No. of bitstreams: 1 347982.pdf: 1600667 bytes, checksum: 2a60ad3de906ac67f5a05683369e9a69 (MD5) Previous issue date: 2006-06-07 / A import?ncia da intera??o entre o mecanismo de coagula??o e o c?ncer ? reconhecida desde as descri??es pioneiras de Trousseau. Mais recentemente, estudos demonstraram a import?ncia do fator tecidual, uma prote?na de membrana celular ligada ao desencadeamento da cascata da coagula??o, na progress?o de neoplasias malignas atrav?s da estimula??o do processo de angiog?nese. Realizamos marca??o imunoistoqu?mica para o fator tecidual em 43 adenocarcinomas colo-retais de pacientes submetidos a tratamento cir?rgico no Servi?o de Coloproctologia do HSL-PUCRS e comparamos a intensidade da sua express?o com a densidade microvascular, crit?rios de estadiamento, idade, sexo e sobrevida geral. Oitenta e tr?s por cento dos tumores apresentaram alta express?o do fator tecidual, havendo uma rela??o estatisticamente significativa entre esta e a maior densidade microvascular (p=0,02). Observamos tamb?m que os pacientes com alta express?o do fator tecidual apresentavam uma m?dia de idade significativamente maior do que a dos pacientes com baixa express?o desta prote?na (p<0,01). Em conclus?o, a elevada intensidade de marca??o imunoistoqu?mica para o fator tecidual se relacionou com uma m?dia de densidade microvascular mais elevada e com pacientes mais idosos quando comparados aos casos com baixa express?o desta prote?na.

Made available in DSpace on 2015-04-14T13:35:52Z (GMT). No. of bitstreams: 1 455926.pdf: 1580649 bytes, checksum: 43ccab4db0f539ea4361b47f57d0cedd (MD5) Previous issue date: 2014-01-24 / We assessed the effects of pharmacological spinal inhibition of voltage-gated calcium channels (VGCC) in mouse pruritus. The epidural administration of P/Q-type MVIIC or PhTx3.3, L-type verapamil, T-type NNC 55-0396 or R-type SNX-482 VGCC blockers failed to alter the scratching behavior caused by the PAR-2 activator trypsin, injected into the mouse nape skin. Otherwise, trypsin-elicited pruritus was markedly reduced by the spinal administration of preferential N-type VGCC inhibitors MVIIA and Ph&#945;1&#946;. C. magus-obtained toxin MVIIA displayed significant effects when dosed from 1 to 4 h before trypsin, whereas the effects of P. nigriventer-derived Ph&#945;1&#946; remained for up to 12 h. MVIIA or Ph&#945;1&#946; also prevented the itching elicited by intradermal (i.d.) injection of SLIGRL-NH2, compound 48/80 or chloroquine, although they did not affect H2O2-induced itching. Furthermore, the co-administration of MVIIA or Ph&#945;1&#946; markedly inhibited the pruritus caused by the spinal injection of gastrin-releasing peptide (GRP), but not morphine. Notably, spinal MVIIA or Ph&#945;1&#946; greatly prevented chronic pruritus allied to dry skin. However, either toxin failed to alter the edema formation or neutrophil influx caused by trypsin. In addition, epidural MVIIA or Ph&#945;1&#946; did not modify the expression of GRP receptor (GRP-R) in the spinal cord, whilst they brought c-Fos activation to control levels. Finally, the in vitro incubation of MVIIA or Ph&#945;1&#946; prevented the calcium influx evoked by the synthetic PAR-2 agonist AC264613 in spinal cord synaptosomes. Data brings novel evidence on itching transmission mechanisms, pointing out the therapeutic relevance of N-type VGCC inhibitors to control refractory pruritus. / Foram avaliados os efeitos da inibi??o farmacol?gica dos canais de c?lcio voltagem-dependentes (CCVD) no prurido em camundongos. A administra??o intratecal (i.t.) dos bloqueadores CCVD do subtipo P/Q (MVIIA ou PhTx3.3), subtipo L (verapamil), subtipo T (NNC 55-0396) ou do subtipo R (SNX-482) n?o alterou o comportamento de co?ar induzido pela tripsina (agonista do receptor PAR-2), aplicada na regi?o dorsal do pesco?o de camundongos. Entretanto, o comportamento de co?ar induzido pela tripsina foi significativamente diminu?do pela administra??o i.t. dos bloqueadores de CCVD do subtipo N (MVIIA e Ph&#945;1&#946;). A toxina MVIIA, derivada do C. magus, apresentou efeitos significativos quando administrada entre 1 a 4 horas antes da aplica??o de tripsina. Por outro lado, a toxina Ph&#945;1&#946; derivada do veneno da aranha P. nigriventer demonstrou atividade, quando injetada at? 12 h antes da tripsina. O pr?-tratamento com MVIIA ou Ph&#945;1&#946; foi efetivo em inibir o comportamento de co?ar induzido pela aplica??o intrad?rmica (i.d.) de SLIGRL-NH2, composto 48/80 ou cloroquina; entretanto, as toxinas n?o inibiram o comportamento de co?ar induzido pela inje??o i.d. de H2O2. A co-inje??o de MVIIA ou Ph&#945;1&#946; inibiu o comportamento de co?ar induzido pela aplica??o i.t. do pept?deo liberador de gastrina (GRP), mas n?o inibiu o prurido causado pela morfina. Relevantemente, a administra??o i.t. da toxina MVIIA ou Ph&#945;1&#946; inibiu a coceira cr?nica induzida pelo modelo de pele seca em camundongos. A atividade anti-pruritog?nica de ambas as toxinas n?o parece estar relacionada com a modula??o do processo inflamat?rio na pele, uma vez que as toxinas MVIIA ou Ph&#945;1&#946; n?o foram capazes de inibir o edema e a migra??o de neutr?filos induzidos pela aplica??o i.d. de tripsina. A aplica??o i.t. de MVIIA ou Ph&#945;1&#946; n?o modificou a express?o do receptor para o GRP (GRP-R) na medula espinhal. Por outro lado, as duas reduziram a express?o de c-Fos aos n?veis do grupo controle, de acordo com a avalia??o na medula dos camundongos. Finalmente, a incuba??o de MVIIA ou Ph&#945;1&#946; preveniu o influxo de c?lcio estimulado pelo agonista do receptor PAR-2 AC264613 em sinaptossomas de medula de camundongos. Estes resultados trazem uma nova perspectiva acerca dos mecanismos envolvidos na sinaliza??o da coceira, indicando os inibidores de CCVD do subtipo N como poss?veis estrat?gias para o tratamento do prurido, especialmente nas condi??es onde h? aus?ncia de resposta ? terapia atual.

Submitted by Setor de Tratamento da Informa??o - BC/PUCRS (tede2@pucrs.br) on 2016-09-26T11:17:54Z No. of bitstreams: 1 DIS_JOSE_JACQUES_GARCEZ_COMPLETO.pdf: 2188213 bytes, checksum: 52d9379f9b56a2cbd55ff55765b34197 (MD5) / Made available in DSpace on 2016-09-26T11:17:54Z (GMT). No. of bitstreams: 1 DIS_JOSE_JACQUES_GARCEZ_COMPLETO.pdf: 2188213 bytes, checksum: 52d9379f9b56a2cbd55ff55765b34197 (MD5) Previous issue date: 2016-08-03 / This master thesis aimed the study of different extraction techniques and the influence of different particle sizes in maximizing the yield of essential oil from the seeds of dill, an herb widely used in cooking and in traditional folk medicine and has a wide range of properties conferred such as antimicrobial, antibacterial and antioxidant. The techniques used for obtaining the essential oil were extracted by steam distillation and hydrodistillation, where different average particle sizes of crushed seed and anise were considered, and extraction with supercritical carbon dioxide, where besides the variation of average particle sizes it was used different pressure values. For the supercritical extraction, it was then used the response surface method in which the variables pressure and average particle diameter were investigated, also a 2? factorial design was used to determine the number of experiments required. The best conditions for the three extractive methods were found, being larger diameters of particles to steam distillation and hydrodistillation, and 10,000 kPa and smaller diameter particles to the supercritical extraction. In this evaluation, it was performed mathematical modeling of the yield versus time curves of the best extraction conditions for each of the techniques. The extracts were analyzed in a gas chromatograph coupled to a mass spectrometer, after, the results have undergone a statistical analysis of major components, which were divided into 3 different groups according to their similar compositions. Finally, its antibacterial activities were tested where all extracts showed activity against E. coli. / Este trabalho tem como objetivo o estudo sobre algumas diferentes t?cnicas de extra??o e a influ?ncia de diferentes granulometrias na maximiza??o do rendimento do ?leo essencial das sementes do endro, uma erva utilizada na culin?ria e na medicina popular tradicional e que possui uma ampla gama de propriedades atribu?das tais como atividade antibacteriana, antif?ngica e antioxidante. As t?cnicas realizadas para a obten??o do ?leo essencial foram: extra??o por arraste a vapor e hidrodestila??o, onde diferentes tamanhos m?dios das part?culas de sementes mo?das do endro foram considerados, e extra??o com di?xido de carbono supercr?tico, onde al?m da varia??o dos tamanhos m?dios das part?culas, foram empregados diferentes valores de press?es. Para a extra??o supercr?tica, foi ent?o utilizado o m?todo de superf?cie de respostas onde as vari?veis, press?o e di?metro m?dio de part?culas foram investigados, tamb?m, um planejamento fatorial 2? foi aplicado para determinar a quantidade de experimentos necess?rios. As melhores condi??es nos tr?s m?todos extrativos foram encontradas, sendo maiores di?metros de part?culas para o arraste a vapor e a hidrodestila??o e 10.000 kPa e menor di?metro de part?culas para a extra??o supercr?tica. Al?m desta avalia??o, foram realizadas modelagens matem?ticas das curvas de rendimento versus tempo das melhores condi??es de extra??o referente a cada uma das t?cnicas. Os extratos foram analisados em um cromat?grafo a g?s acoplado a um espectr?metro de massas, ap?s, os resultados passaram por uma an?lise estat?stica de componentes principais, onde foram divididos em tr?s diferentes grupos de acordo com suas semelhantes composi??es. Por fim, suas a??es antibacterianas foram testadas onde todos os extratos apresentaram atividade frente a E. coli.

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2016-10-26T18:07:29Z No. of bitstreams: 1 2015 - Ileana Costa Miranda.pdf: 4377950 bytes, checksum: 0fd81889ff042fdd5ef81568986b1e7a (MD5) / Made available in DSpace on 2016-10-26T18:07:29Z (GMT). No. of bitstreams: 1 2015 - Ileana Costa Miranda.pdf: 4377950 bytes, checksum: 0fd81889ff042fdd5ef81568986b1e7a (MD5) Previous issue date: 2015-12-15 / CAPES / Animals grazing Brachiaria spp commonly present foamy macrophages isolated or grouped in the liver, and crystals within biliary ducts. The pathogenesis of formation and the nature of the material stored on these cells, however, are not completely known. Through lectin histochemistry evaluation, steroidal saponins (secondary glycosylated metabolites) have been identified in the crystals and within the cytoplasm of the foam cells, which are probably liable for damage the liver leading to accumulation of phylloerythrin. This study aims to standardize the use of lectin histochemistry to detect glycosylated metabolites in tissues of buffaloes kept in different Brachiaria spp pastures in Brazil. Fragments of liver and mesenteric lymph node from 40 animals were analyzed: 10 buffaloes that were kept in predominant pasture of B. decumbens for 12 months; 10 buffaloes that were kept in pasture with a predominance of B. brizantha for 18 months; 10 buffaloes that were kept in pasture of B. brizantha for approximately four years; and, as a negative control, 10 buffaloes that were maintained in native pasture without Brachiaria spp since birth. Fourteen lectins were tested (Con-A, SBA, WGA, DBA, UEA, RCA, PNA, GSL-I, PSA, LCA, PHA-E, PHA-L, SJA and SWGA), in a total of 1120 evaluated fragments. Previous studies demonstrated that PNA showed great binding reactivity for foamy macrophages in bovine and ovine. In the present study, SWGA presented high specificity and marked binding reactivity for foamy macrophages; WGA, GSL, PHA-E and PHA-L showed moderate to marked reactivity but low specificity for foamy macrophages; the other lectins didn't show significant reactivity or specificity. It remains unclear why there is this difference in lectins binding reactivity to foamy macrophages; it is suggested that divergences may occur depending on the species of Brachiaria ingested, the plant growth stage, the type and proportion of saponins stored in the plant due to seasonality, the differences in the metabolism of animal species, the presence of photosensitivity, the clinical course of the disease and the plant intake time. Moreover, there was no significant reactivity difference between the collected fragments of animals that grazed in B. decumbens for 12 months and B. brizantha for 18 months. However, the decreased presence of foamy macrophages and its lectin histochemical binding in animals that fed on B. brizantha for a longer time indicates that the animals can pass through an adaptation process according to the the plant intake time. Lectin histochemistry analysis can be used to characterize the material stored in foamy macrophages present in liver and mesenteric lymph node of buffaloes that graze in Brachiaria spp pastures and helps to clarify the pathogenesis of these cells. / Animais que se alimentam em pastos de Brachiaria spp comumente apresentam macr?fagos espumosos isolados ou agrupados no f?gado, al?m de cristais no interior de ductos biliares. A patog?nese da forma??o e a natureza do material armazenado nestas c?lulas, contudo, ainda n?o s?o completamente conhecidas. Atrav?s da avalia??o lectino-histoqu?mica, saponinas esteroidais (metab?litos glicosilados secund?rios) t?m sido identificadas nos cristais e no citoplasma das c?lulas espumosas, que provavelmente s?o respons?veis por danificar o f?gado e levar ao ac?mulo de filoeritrina. Por meio deste trabalho, objetivou-se padronizar e caracterizar a utiliza??o da lectino-histoqu?mica na detec??o de metab?litos glicosilados nos tecidos de b?falos mantidos em diferentes pastos de Brachiaria spp no Brasil. Fragmentos de f?gado e linfonodo mesent?rico de 40 animais foram analisados: 10 b?falos mantidos em pastagem predominante de B. decumbens por aproximadamente 12 meses; 10 b?falos mantidos em pastagem predominante de B. brizantha por aproximadamente 18 meses; 10 b?falos mantidos em pastagem de B. brizantha por aproximadamente quatro anos; e, como controle negativo, 10 b?falos mantidos em pastagem livre de Brachiaria spp desde o nascimento. Quatorze lectinas foram testadas (Con-A, SBA, WGA, DBA, UEA, RCA, PNA, GSL-I, PSA, LCA, PHA-E, PHA-L, SJA e SWGA), em um total de 1120 fragmentos avaliados. Estudos anteriores demonstraram que a lectina PNA possui marcada reatividade para macr?fagos espumosos de bovinos e ovinos. No presente estudo, a lectina SWGA apresentou acentuada reatividade e alta especificidade para macr?fagos espumosos; WGA, GSL, PHA-E e PHA-L mostraram moderada a acentuada reatividade, mas baixa especificidade aos macr?fagos espumosos; as outras lectinas n?o apresentaram reatividade ou especificidade significativas. Ainda n?o se sabe exatamente a que atribuir a diferen?a de reatividade aos macr?fagos espumosos. Sugere-se que diverg?ncias ocorram em fun??o da esp?cie de Brachiaria ingerida, da fase de crescimento da planta, do tipo e propor??o dos glicoconjugados armazenados na planta em decorr?ncia da ?poca do ano, das diferen?as no metabolismo da esp?cie do animal em quest?o, da presen?a de fotossensibiliza??o, da evolu??o cl?nica da doen?a e do tempo de ingest?o da planta. N?o houve diferen?a de marca??o significativa entre os fragmentos coletados de animais que se alimentaram de B. decumbens por 12 meses e B. brizantha por 18 meses. Por?m, a diminui??o da presen?a e marca??o lectino-histoqu?mica dos macr?fagos espumosos nos tecidos dos b?falos que ingeriram B. brizantha durante mais tempo indica que os animais podem passar por um processo de adapta??o de acordo com o tempo de ingest?o da planta. A avalia??o lectino-histoqu?mica pode ser utilizada para caracterizar o material armazenado em macr?fagos espumosos presentes no f?gado e linfonodo mesent?rico de b?falos que se alimentam em pastagens de Brachiaria spp e ajuda na compreens?o da patog?nese de forma??o destas c?lulas.

Submitted by Setor de Tratamento da Informa??o - BC/PUCRS (tede2@pucrs.br) on 2017-03-10T15:09:55Z No. of bitstreams: 1 TES_JULIANO_SARTORI_COMPLETO.pdf: 9083435 bytes, checksum: 6cadde5ff4cace9cc886d4131547c873 (MD5) / Made available in DSpace on 2017-03-10T15:09:55Z (GMT). No. of bitstreams: 1 TES_JULIANO_SARTORI_COMPLETO.pdf: 9083435 bytes, checksum: 6cadde5ff4cace9cc886d4131547c873 (MD5) Previous issue date: 2016-12-02 / Despite the advances resulting from the molecular characterization of breast cancer that allowed the use of more selective therapeutic targets, the disease still causes different clinical outcomes with high rates of recurrence and mortality. In this context of complexity and heterogeneity of breast cancer, are important the investigations of new biomolecular markers related to breast oncogenesis that may contribute to know the prognosis and improve the clinical management of patients. The sirtuin-1 (SIRT-1) is a histone deacetylase implicated in various epigenetic critical functions into cells, among these, the maintenance of genomic stability, proliferation and cell aging. The aim of this study was to investigate the grade of expression of the sirtuin- 1 (SIRT-1) in a cohort of 457 women with breast cancer and verify the effect, independent or in combination with other variables in the prognosis of these patients. It is a survival analysis study based on hospital medical records of women with breast cancer undergoing treatment in Erechim-RS from 2003 to 2013 and followed until 31 July 2015. The analysis of the grade of SIRT-1 expression was performed by immunohistochemistry in 123 patients (26.9%) of the total cohort. Overall survival specific disease (OS) was estimated by the Kaplan-Meier method and the risk of death from breast cancer by the method of Cox proportional hazards. The research was approved by the Research Ethics Committee of PUC-RS as reported number 465.362. The median age was 57.4 years and the median estimate of breast cancer survival was 79.6% at 5 years and 69.1% at 10 years, with median follow-up time of 61.9 months. Risk factors associated with worse prognosis were: age between 60 and 69 years (HR = 1.88; 95% CI 1.02 - 3.44; p = 0.042); older than 70 years (HR = 2.92; CI 95% 1.70 - 5.01; p = 0.000); tumor size greater than 2 cm (HR = 1.86; CI 95% 1.04 -3.36; p = 0.038); metastasis in 4 or more axillary lymph nodes (HR = 2.37; CI 95% 1.38 - 4.08; p = 0.002); located clinical staging (CS II, TNM) (HR = 3.39; CI 95% 1.35 - 8.51; p = 0.009); advanced clinical staging (CS III, TNM) (HR = 6.32; CI 95% 2.49-16.08; p = 0.000); high histologic grade (HR = 6.32; CI 95% 1.63 - 29.99; p = 0.008); triple negative molecular profile (HR = 2.33; CI 95% 1.19 - 4.58; p = 0.014) and radical surgery (HR = 2.10; CI 95% 1.31 - 3.36; p = 0.002). The positive expression of progesterone receptor (HR = 0.52, CI 95% 0.34 - 0.79; p = 0.002) was a better prognostic factor for patients. The grade of overexpression of SIRT-1, defined as nuclear expression of SIRT-1 greater than 80% was observed in 6.5% of cases. The SIRT-1 overexpression characterized a subgroup of women who had a worse prognosis with shorter survival and increased risk of death from breast cancer (HR = 2.66; CI 95% 1.03 - 6.86; p = 0.043). Multivariate regressive models (Cox) were constructed and the overexpression of SIRT-1 remained significant statistic demonstrating independent factor associated with worse prognostic in breast cancer. Therefore, the evaluation of the grade of expression of the SIRT-1, in the cohort of Erechim-RS, proved to be an independent prognostic marker for analysis of the risk of death from breast cancer. / Apesar dos avan?os decorrentes da caracteriza??o molecular do c?ncer de mama que permitiram o emprego de alvos terap?uticos mais seletivos, a doen?a ainda ocasiona diferentes desfechos cl?nicos, com elevadas taxas de recidiva e mortalidade. Neste contexto de complexidade e heterogeneidade do c?ncer de mama, s?o importantes as investiga??es de novos marcadores biomoleculares relacionados ? oncog?nese mam?ria que possam contribuir para conhecer o progn?stico e aprimorar o manejo cl?nico das pacientes. A Sirtu?na-1 (SIRT-1) ? uma histona desacetilase implicada em diversas fun??es epigen?ticas cr?ticas para as c?lulas, dentre estas, a manuten??o da estabilidade gen?mica, a prolifera??o e o envelhecimento celular. O objetivo deste estudo foi investigar o grau de express?o da Sirtu?na-1 (SIRT-1) em uma coorte de 457 mulheres portadoras de c?ncer de mama e verificar o seu efeito, independente ou em associa??o a outras vari?veis, no progn?stico destas pacientes. Trata-se de um estudo de an?lise de sobrevida com base em registros de mulheres portadoras de c?ncer de mama submetidas a tratamento em Erechim-RS no per?odo de 2003 a 2013 com seguimento at? 31 de julho de 2015. A an?lise do grau de express?o de SIRT-1 foi realizada por t?cnica de imuno-histoqu?mica em 123 pacientes (26,9%) do total da coorte. A sobrevida global doen?a espec?fica foi estimada pelo m?todo de Kaplan-Meier e, o risco de morte, por c?ncer de mama, pelo m?todo de riscos proporcionais de Cox. A pesquisa foi aprovada no Comit? de ?tica e Pesquisa da PUC-RS, sob n? 465.362. A idade mediana foi de 57,4 anos e a estimativa mediana de sobrevida por c?ncer de mama foi de 79,6% em 5 anos e 69,1% em 10 anos, com tempo mediano de seguimento de 61,9 meses. Os fatores de risco associados a pior progn?stico foram: faixa et?ria entre 60 e 69 anos (HR=1,88; IC95% 1,02-3,44; p=0,042); faixa et?ria maior que 70 anos (HR=2,92; IC95% 1,70-5,01; p=0,000); tamanho tumoral acima de 2 cm (HR=1,86; IC95% 1,04-3,36; p=0,038); presen?a de met?stase em 4 linfonodos axilares ou mais (HR=2,37; IC95% 1,38-4,08; p=0,002); estadiamento cl?nico localizado (EC II,TNM) (HR=3,39; IC95% 1,35-8,51; p=0,009); estadiamento cl?nico avan?ado (EC III,TNM) (HR=6,32; IC95% 2,49-16,08; p=0,000); grau histol?gico alto (HR=6,32; IC95% 1,63-29,99; p=0,008); perfil molecular triplo negativo (HR=2,33; IC95% 1,19-4,58; p=0,014) e a cirurgia radical (HR=2,10; IC95% 1,31-3,36; p=0,002). A express?o positiva do receptor de progesterona (HR=0,52; IC95% 0,34- 0,79; p=0,002) foi um fator de melhor progn?stico para as pacientes. O grau de hiperexpress?o da SIRT-1, definida como express?o nuclear da SIRT-1 maior que 80%, foi verificada em 6,5% dos casos. A hiperexpress?o da SIRT-1 caracterizou um subgrupo de mulheres que apresentaram pior progn?stico, com menor sobrevida e maior risco de morte por c?ncer de mama (HR=2,66; IC95% 1,03-6,86; p=0,043). Foram elaborados modelos regressivos multivariados (Cox) e a hiperexpress?o de SIRT-1 manteve signific?ncia estat?stica demonstrando fator independente associado a pior progn?stico no c?ncer de mama. Portanto, a avalia??o do grau de express?o da SIRT-1, na coorte de Erechim-RS, demonstrou ser um marcador independente para determinar o progn?stico no c?ncer de mama.

Submitted by Sara Ribeiro (sara.ribeiro@ucb.br) on 2017-06-02T12:47:54Z No. of bitstreams: 1 MarianaRochaMaximianoDissertacaoParcial2015.pdf: 459398 bytes, checksum: b2185a3a3375dbb7ac1ed82fc658146b (MD5) / Approved for entry into archive by Sara Ribeiro (sara.ribeiro@ucb.br) on 2017-06-02T12:48:21Z (GMT) No. of bitstreams: 1 MarianaRochaMaximianoDissertacaoParcial2015.pdf: 459398 bytes, checksum: b2185a3a3375dbb7ac1ed82fc658146b (MD5) / Made available in DSpace on 2017-06-02T12:48:21Z (GMT). No. of bitstreams: 1 MarianaRochaMaximianoDissertacaoParcial2015.pdf: 459398 bytes, checksum: b2185a3a3375dbb7ac1ed82fc658146b (MD5) Previous issue date: 2015-06-18 / Sclerotinia sclerotiorum, the causal agent of white mold, is a plant pathogenic fungus that has a characteristic feature, production of sclerotia, resistance structures that can be viable for up to 10 years in the soil, with ability to initiate a new cycle of infection under favorable conditions. The disease control methods are based on integrated management practices, including biological and chemical control. The proteomic study of this interaction may be a strategy for studying this pathosystem, however, the small amount of mycelium produced during the infectious process in vivo greatly limits the study of this pathosystem using this strategy. The main goals of this work were the development and validation of a culture medium that would partially mimic the host and allow the production of large amounts of micelium for in vitro studies of this pathosystem. For this purpose, a protocol was established for the production of culture media prepared with leaf extract of the hosts. These media were inoculated with sclerotia of the monosporic isolate SS 200 of S. sclerotiorum and the differential expression of effectors and candidate effectors of virulence of the fungus was evaluated by qPCR. The results showed that a large amount of micelia grew in the media and effector genes and candidate effector genes were induced in these media. These results indicate that the proposed culture media can be used to study the in vitro interaction between S. sclerotiorum and several plant hosts and that they can be useful especially in proteomic studies. / Sclerotinia sclerotiorum, agente causal do mofo branco, ?? um fungo fitopatog??nico que possui uma caracter??stica marcante, a produ????o de escler??dios, estruturas de resist??ncia que podem ser vi??veis por at?? 10 anos no solo, com capacidade de iniciar um novo ciclo de infec????o em condi????es favor??veis. Os m??todos de controle da doen??a baseiam-se em pr??ticas de manejo integrado, incluindo controle biol??gico e qu??mico. O estudo prote??mico desta intera????o pode ser uma estrat??gia para o estudo deste patossistema, entretanto, a pequena quantidade de mic??lio produzido pelo pat??geno durante o processo infeccioso in vivo limita bastante o estudo deste patossistema utilizando esta estrat??gia. Os objetivos deste trabalho foram o desenvolvimento e valida????o de um meio de cultura que mimetizasse parcialmente o hospedeiro e permitisse a produ????o de grande quantidade de massa micelial para estudos in vitro desse patossistema. Para tal, foi estabelecido um protocolo para a produ????o de meios de cultura, preparados a partir de extratos foliares dos hospedeiros. Estes meios foram inoculados com escler??dios do isolado monosp??rico de S. sclerotiorum SS 200 e a express??o diferencial de genes efetores e candidatos a efetores de virul??ncia do fungo foi avaliada por qPCR. Os resultados mostraram que em todos os meios testados houve crescimento de grande quantidade de massa micelial e que os meios foram capazes de induzir a express??o de genes efetores e candidatos a efetores. Portanto, os resultados indicam que os meios propostos podem ser usados no estudo da intera????o in vitro de S. sclerotiorum com v??rios hospedeiros, especialmente em estudos prote??micos.