MiRNAs são pequenas moléculas de RNAs endógenos não codificantes com aproximadamente 23nt que atuam na regulação da expressão gênica. A sua função é inibir a tradução de genes transcritos através de um mecanismo que viabiliza a ligação do miRNA com o mRNA alvo levando à inibição da tradução ou a degradação do RNA mensageiro. Estudos evidenciam a relação dos miRNAs com diversos processos biológicos como proliferação celular, diferenciação, desenvolvimento e doenças. Uma vez que estão envolvidos na regulação gênica, também alteram as vias metabólicas. Atualmente, as ferramentas computacionais disponíveis para o estudo dos miRNAs são o miRBase, microCosm, o miRGen e o miRNAmap. Elas possuem informações sobre as sequências dos miRNAs, genes alvos e sobre elementos que estão próximos à região dos miRNAs. Embora o avanço até o momento, não existia que relacionasse os miRNAs com as vias metabólicas, para isso foi construída a plataforma miRNApath que auxilia no estudo da função dos miRNAs por meio da análise do seus alvos dentro vias metabólicas. De modo semelhante, também não existia uma abordagem que relacione dados de expressão miRNAs e seus alvos dentro de um mesmo experimento. Para tanto, neste trabalho foi feita uma abordagem utilizando bibliotecas de SAGE (Serial Analysis of Gene Expression) que será incorporada no miRNApath. O miRNApath encontra-se disponível em http://lgmb.fmrp.usp.br/mirnapath. / MiRNAs are small molecules of endogenous non-coding RNAs with approximately 23nt in length that acts over gene expression regulation. Its function is inhibit the translation of gene transcripts through a mechanism that links the miRNA with its mRNA target leading to a translational repression or degradation. Studies show the relation of RNAs in many biological processes like cell proliferation, dierentiation and development of diseases. Since they are involved in gene regulation, they also change the metabolic pathways. Currently, the available computational tools for the study of miRNAs are miRBase, microCosm, miRGen and miRNAmap. They have information about miRNAs sequences, targets and features. Despite the the advances, until now, there is no tool that correlates the miRNAs with metabolic pathways, therefore we developed the miRNApath platform that helps in the analysis of miRNAs function through the study of its targets that are into the metabolic pathway. In the same way, there is no approach that put together information of expression of miRNAs and its targets in the same experiment. In this work we develop an approach with SAGE (Serial Analysis of Gene Expression ) libraries that will be integrated to miRNApath. The plataform is avaible at http://lgmb.fmrp.usp.br/mirnapath.

MALTA, T.M. Análise do perfil de expressão gênica em linfócitos T CD8+ isolados de pacientes infectados pelo HTLV-1. 2009. 137f. Dissertação de Mestrado. Faculdade de Ciências Farmacêuticas de Ribeirão Preto - Universidade de São Paulo, Ribeirão Preto, 2009. O vírus linfotrópico de células T humanas tipo 1 (HTLV-1) foi o primeiro retrovírus humano descrito e está etiologicamente associado a duas principais manifestações clínicas: a leucemia ou linfoma de células T do adulto (ATLL) e a mielopatia associada ao HTLV-1 ou paraparesia espástica tropical (HAM/TSP). É estimado que 3 a 5% das pessoas infectadas pelo HTLV-1 desenvolvem as doenças associadas, enquanto a maioria permanece assintomática. A HAM/TSP é uma manifestação inflamatória do sistema nervoso central e o mecanismo pelo qual o HTLV-1 induz o surgimento de HAM/TSP ainda é questão de debate. Os linfócitos T CD8+ citotóxicos (CTL) têm uma participação importante na resposta imunológica dirigida contra o HTLV-1 e a eficiência com que as CTLs eliminam as células infectadas pelo vírus está relacionada com a carga proviral (CPV) do indivíduo e consequentemente com o risco de desenvolvimento de HAM/TSP. No presente trabalho, foram avaliados os perfis de expressão gênica de linfócitos T CD8+ isolados de portadores assintomáticos (HAC), pacientes com HAM/TSP (HAM/TSP) e de indivíduos sadios (CT) por meio da metodologia de Serial Analysis of Gene Expression (SAGE). Os linfócitos T CD8+ utilizados foram isolados por meio de separação imunomagnética e um pool composto de quatro indivíduos infectados para cada grupo, HAC e HAM/TSP foi utilizado para a construção das bibliotecas. A análise do SAGE resultou num total de 51.017, 62.432 e 60.620 tags sequenciadas para as bibliotecas CT, HAC e HAM/TSP, respectivamente, o que permitiu a identificação de aproximadamente 12.000 transcritos diferentes em cada biblioteca. Foram identificados cerca de 900 genes diferencialmente expressos entre as bibliotecas CT e HAC ou HAM/TSP. A comparação dos grupos HAC e HAM/TSP revelou 290 genes. Um grande número destes genes diferencialmente expressos está envolvido com os processos de apoptose, adesão e migração celular. Os achados foram validados pela metodologia de PCR em tempo real em um total de 17 indivíduos assintomáticos, 14 com HAM/TSP e 24 indivíduos sadios. A validação evidenciou o aumento da expressão dos genes envolvidos na lise mediada por células PRF1, GZMB e GZMH, além do aumento de CCL5, ZAP70 e PXN nos indivíduos infectados pelo HTLV-1. Além disso, os resultados mostraram redução nos níveis de expressão de CXCR4 nesses pacientes. Adicionalmente, foi realizada a quantificação dos níveis protéicos de PRF1 e GZMB por citometria de fluxo e os resultados corroboraram com a PCR em tempo real. O perfil gênico dos linfócitos T CD8+ demonstrou a ocorrência de intensa ativação e migração celular durante a infecção pelo HTLV-1. Essas evidências indicam que as células CD8+ dos indivíduos infectados pelo HTLV-1 parecem contribuir mais para a proteção e controle da infecção pelo vírus, e menos para o desenvolvimento de HAM/TSP. Este foi o primeiro trabalho a analisar a expressão global de linfócitos T CD8+ nesta infecção por meio de SAGE e permitiu a geração de dados que serão empregados em diferentes abordagens na pesquisa com HTLV-1. / MALTA, T.M. Serial Analysis of Gene Expression in CD8+ T-cells isolated from HTLV-1 infected individuals. 2009. 137f. Dissertação de Mestrado. Faculdade de Ciências Farmacêuticas de Ribeirão Preto - Universidade de São Paulo, Ribeirão Preto, 2009. Human T-cell lymphotropic virus type 1 (HTLV-1) was the first human retrovirus discovered and is related with two major diseases: adult T cell lymphoma/leukaemia (ATLL) and HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP). About 3 to 5% of infected individuals will develop HTLV-1 related diseases, while the majority remains life-long asymptomatic carriers of the virus. HAM/TSP is an inflammatory manifestation of central nervous system and the mechanism involved in HAM/TSP development are not well elucidated. CD8+ cytotoxic T lymphocytes (CTL) have an important function in the immune response against the HTLV-1 and is related with the individual proviral load and so in the risk of HAM/TSP. To identify genes differentially expressed among non-infected individuals, asymptomatic (HAC) and HAM/TSP (HAM/TSP) patients we performed a serial analysis of gene expression (SAGE) using CD8+ T cells isolated from these individuals. The CD8+ T lymphocytes were isolated by magnetic cell separation system and HAC and HAM/TSP SAGE libraries were composed by pooled of four samples. SAGE analysis of 51,017, 62,432 and 60,620 tags from control, HAC and HAM/TSP groups respectively, allowed identification of approximately 12,000 different transcripts in each library. The expression profile revealed around 900 genes differentially expressed between control group and HAC or HAM/TSP, and 290 genes were identified between HAC and HAM/TSP groups. The expression profile revealed the presence of highly frequent transcripts related to apoptosis, cell adhesion and cell migration. These results were validated by real time PCR in 17 HTLV-1 asymptomatic carriers, 14 patients with HAM/TSP and 24 healthy individuals. The validation revealed the increased levels of expression of the cytolysis genes PRF1, GZMB and GZMH, and the increases of CCL5, ZAP70 and PXN in the HTLV-1 infected individuals. Besides, the CXCR4 was suppressed in these patients. Additionally, we performed the protein quantification of PRF1 and GZMB by flow cytometry and the real time PCR results were confirmed. The CD8+ T cells profiles showed strong cell activation and cell migration in HTLV-1 infection. These data suggests that CD8+ T cells from HTLV-1 individuals seem to contribute more to the protection and virus infection control than to the HAM/TSP development. This study represents the first extensive serial analysis of gene expression of CD8+T cells in HTLV-1 infection and allowed to generate data that will be used in different approaches in HTLV-1 research.

No description available.

No description available.

Management of greater sage-grouse (Centrocercus urophasianus) in the west has changed over the last several decades in response to environmental and anthropogenic causes. Many land and wildlife management agencies have begun manipulating sagebrush with herbicides, machinery, and fire. The intent of these manipulations (treatments) is to reduce sagebrush canopy cover and increase the density of grass and forb species, thus providing higher quality sage-grouse brood-rearing habitat. However, monitoring of sage-grouse response to such manipulations has often been lacking or non-existent. The objective of our study was to determine the response of sage-grouse to sagebrush reduction treatments that have occurred recently in Rich County, Utah. Our study areas were treated with a pasture aerator with the intent of creating sage-grouse brood-rearing habitat. We used pellet transects, occupancy sampling, and GPS radio telemetry to quantify sage-grouse habitat use in treated and untreated areas. Pellet transect, occupancy, and GPS radio telemetry methods all showed a strong pattern of sage-grouse use of treated sites during the breeding and early brood-rearing periods. Sage-grouse use of treated sites was greatest in lower elevation habitat (1950 to 2110 m), and use was highest during the breeding and early brood-rearing periods. We found very little use of higher elevation (2120 to 2250 m) treated or untreated sites. Our results suggest that sagebrush reduction treatments can have positive impacts on sage-grouse use at lower elevations and can be successful in creating brood-rearing habitat. Elevation differences and period of sage-grouse use were significant factors in our study in determining how beneficial sagebrush reduction treatments were for sage-grouse.

En traitant de la recherche personnelle de l'utile propre des individus, Spinoza semble s'inscrire dans une tradition philosophique égoïste, où l'intérêt pour soi empêche d'avoir un véritable souci pour l'autre. Pourtant, cette lecture de Spinoza entre en contradiction avec l'idée de générosité, generositas , telle que décrite au scolie de la proposition 59 de la troisième partie de l'Éthique. "Par générosité, j'entends un Désir par lequel un individu s'efforce en vertu du seul commandement de la raison à assister les autres hommes et à établir entre eux et lui un lien d'amitié. Je rapporte donc à la fermeté ces actions qui ont pour but l'utilité de l'agent seulement, et à la générosité celles qui ont aussi pour but l'utilité d'autrui." Cette définition nous apprend que le sage désire ce qui est utile pour l'autre. Cela pose de nombreuses questions : comment le conatus apparemment égoïste produit-il le désir qui vise l'utilité d'autrui? Comment le désir, individuel d'exister peut-il mécaniquement produire le désir d'aider les autres? Comment la générosité peut-elle venir de la Raison sans pour autant être le fruit d'un calcul égoïste? Quelle forme prendra la générosité spinoziste? Dans ce mémoire, nous avons fait une recherche systématique des occurrences du terme"générosité", recherché les sources de cet affect actif dans l'anthropologie spinoziste en l'opposant à la conception hobbesienne et expliqué comment l'homme généreux interagit avec son environnement.

A single-stage pulse tube cryocooler was designed and fabricated to provide cooling at 50 K for a high temperature superconducting (HTS) magnet, with a nominal electrical input frequency of 50 Hz and a maximum mean helium working gas pressure of 2.5 MPa. Sage software was used for the thermodynamic design of the pulse tube, with an initially predicted 30 W of cooling power at 50 K, and an input indicated power of 1800 W. Sage was found to be a useful tool for the design, and although not perfect, some correlation was established. The fabricated pulse tube was closely coupled to a metallic diaphragm pressure wave generator (PWG) with a 60 ml swept volume. The pulse tube achieved a lowest no-load temperature of 55 K and provided 46 W of cooling power at 77 K with a p-V input power of 675 W, which corresponded to 19.5% of Carnot COP. Recommendations included achieving the specified displacement from the PWG under the higher gas pressures, design and development of a more practical co-axial pulse tube and a multi-stage configuration to achieve the power at lower temperatures required by HTS.

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

The sage grouse or sage hen Centrocercus urophasianus (Bonaparte) was formerly the most important upland native game bird of the Western States, but has steadily been declining in numbers over most of its range in recent years. This reduction in numbers has aroused the interest of the conservationists of the nation, who, for the past decade or more, have been proposing that something be done for this game species. In recent years along with this interest of the conservationists, there has been an increasing local interest among the sportsmen of the West, with the result that several groups have become interested in making a study of the sage grouse and determining reasons for the marked decline in its numbers. Partly because of this aroused interest, the study of the sage grouse was selected as a research project.

Greater sage-grouse (Centrocercus urophasianus; sage-grouse) are sagebrush obligates and are therefore considered to be key indicators of sagebrush ecosystem health. Sage-grouse populations have declined range-wide over the last century due to loss and fragmentation of sagebrush (Artemisia spp.) habitats. Sage-grouse populations found in large intact sagebrush landscapes are considered to be more resilient, however, some small isolated populations persist and thrive in fragmented landscapes. Because of Utah’s unique topography and geography, sage-grouse habitat is discontinuous and populations are naturally dispersed throughout the state in suitable intact blocks or in disconnected islands of sagebrush habitat. Thus, Utah populations provide the ideal place to understand how landscape attributes may influence at risk populations. Of these, the Morgan-Summit population is important because very little was known about the general ecology of this population and it experiences a high level of anthropogenic disturbances. I examined seasonal movement patterns, habitat selection, vital rates (nest initiation rates, nest success, clutch size, breeding success, brood success, and survival probability of breeding age birds) and the influence of vegetation components on vital rates of a small geographically isolated sage-grouse population in Morgan and Summit Counties in northern Utah from 2015–2016. To collect the data, I deployed 25 very-high frequency radio collars and 10 platform terminal transmitters and completed micro-site vegetation surveys at nest, brood, and paired random sites and then made comparisons. Nest sites exhibited variation in vegetation structure that influenced nest success, while brood sites did not. This population is one of the most productive in Utah exhibiting high nest initiation rates, hatching rates, and brood success rates despite limited habitat space and small seasonal movements. Transmitter type had no influence on vital rates, which is contrary to other studies, and limited influence on habitat selection. Sage-grouse avoided trees and developed areas, especially during the breeding season. Selection of other landscape variables was season-dependent. This information suggests that a sage-grouse population can occupy areas of limited habitat on an annual basis if seasonal habitat requirements are met. This study provides information that stake holders can utilize to conserve critical seasonal habitats within this study area where the population could be negatively affected by anthropogenic development pressure.