Human IL-12 p40 as a reporter gene for high-throughput screening of engineered mouse embryonic stem cells

<p>Abstract</p> <p>Background</p> <p>Establishing a suitable level of exogenous gene expression in mammalian cells in general, and embryonic stem (ES) cells in particular, is an important aspect of understanding pathways of cell differentiation, signal transduction and cell physiology. Despite its importance, this process remains challenging because of the poor correlation between the presence of introduced exogenous DNA and its transcription. Consequently, many transfected cells must be screened to identify those with an appropriate level of expression. To improve the screening process, we investigated the utility of the human interleukin 12 (IL-12) p40 cDNA as a reporter gene for studies of mammalian gene expression and for high-throughput screening of engineered mouse embryonic stem cells.</p> <p>Results</p> <p>A series of expression plasmids were used to study the utility of IL-12 p40 as an accurate reporter of gene activity. These studies included a characterization of the IL-12 p40 expression system in terms of: (i) a time course of IL-12 p40 accumulation in the medium of transfected cells; (ii) the dose-response relationship between the input DNA and IL-12 p40 mRNA levels and IL-12 p40 protein secretion; (iii) the utility of IL-12 p40 as a reporter gene for analyzing the activity of <it>cis</it>-acting genetic elements; (iv) expression of the IL-12 p40 reporter protein driven by an IRES element in a bicistronic mRNA; (v) utility of IL-12 p40 as a reporter gene in a high-throughput screening strategy to identify successful transformed mouse embryonic stem cells; (vi) demonstration of pluripotency of IL-12 p40 expressing ES cells <it>in vitro </it>and <it>in vivo</it>; and (vii) germline transmission of the IL-12 p40 reporter gene.</p> <p>Conclusion</p> <p>IL-12 p40 showed several advantages as a reporter gene in terms of sensitivity and ease of the detection procedure. The IL-12 p40 assay was rapid and simple, in as much as the reporter protein secreted from the transfected cells was accurately measured by ELISA using a small aliquot of the culture medium. Remarkably, expression of Il-12 p40 does not affect the pluripotency of mouse ES cells. To our knowledge, human IL-12 p40 is the first secreted reporter protein suitable for high-throughput screening of mouse ES cells. In comparison to other secreted reporters, such as the widely used alkaline phosphatase (SEAP) reporter, the IL-12 p40 reporter system offers other real advantages.</p

Towards a genome-wide transcriptogram: the Saccharomyces cerevisiae case

A genome modular classification that associates cellular processes to modules could lead to a method to quantify the differences in gene expression levels in different cellular stages or conditions: the transcriptogram, a powerful tool for assessing cell performance, would be at hand. Here we present a computational method to order genes on a line that clusters strongly interacting genes, defining functional modules associated with gene ontology terms. The starting point is a list of genes and a matrix specifying their interactions, available at large gene interaction databases. Considering the Saccharomyces cerevisiae genome we produced a succession of plots of gene transcription levels for a fermentation process. These plots discriminate the fermentation stage the cell is going through and may be regarded as the first versions of a transcriptogram. This method is useful for extracting information from cell stimuli/responses experiments, and may be applied with diagnostic purposes to different organisms

Whipple's Disease With Neurological Manifestations: Case Report.

Whipple's disease (WD) is an uncommon multisystem condition caused by the bacillus Tropheryma whipplei. Central nervous system involvement is a classical feature of the disease observed in 20 to 40% of the patients. We report the case of a 62 year old man with WD that developed neurological manifestations during its course, and discuss the most usual signs and symptoms focusing on recent diagnostic criteria and novel treatment regimens.62342-

Avaliação da composição corporal de jogadores de futebol sub-20 da cidade de Uberlândia, Minas Gerais

Introduction: Football depends on several variables to be performed at its highest performance, among these variables, one of the most important is body composition, as it is related to the athlete's physical capacity and sports performance, which can influence the team's results. Objective: To evaluate the body composition of young soccer athletes, in an under-20 team. Methodology: Sixteen soccer players from a sub-20 team, aged 18 to 20 years, were evaluated. Weight and height were measured; in addition, they performed the electrical bioimpedance test to assess the parameters of body composition, mainly muscle mass and fat percentage. From the muscle mass, the muscle mass index (BMI) was calculated, dividing the total muscle mass by height in meters squared. Result: Most athletes were above the ideal fat percentage (n = 12), however, most individuals had values close to the ideal, which may have been a variation of the body composition assessment method itself. We observed that 4 individuals had an adequate percentage of fat. Regarding the IMM, the average values were within the normal range (10.9 ± 0.6 kg / m2), with only 4 athletes having little muscle mass. Conclusion: Almost all athletes had muscle mass within the normal range, while the majority had values above the adequate percentage of fat. These results show that future nutritional interventions should be carried out in these athletes, focusing mainly on reducing the percentage of fat.Trabalho de Conclusão de Curso (Graduação)Introdução: O futebol depende de diversas variáveis para ser realizado no seu mais alto rendimento, entre essas variáveis, uma das mais importantes é a composição corporal, pois está relacionada com a capacidade física e desempenho esportivo do atleta, podendo influenciar nos resultados da equipe. Objetivo: Avaliar a composição corporal de jovens atletas de futebol, em uma equipe sub-20. Metodologia: Foram avaliados 16 atletas de futebol de campo de uma equipe sub-20, com idade de 18 a 20 anos. Foram mensurados peso e estatura; além disso, fizeram o exame de bioimpedância elétrica para avaliar os parâmetros de composição corporal, principalmente massa muscular e percentual de gordura. A partir da massa muscular, foi calculado o índice de massa muscular (IMM), dividindo a massa muscular total pela altura em metros ao quadrado. Resultado: A maioria dos atletas estava acima do percentual de gordura ideal (n= 12), entretanto, a maior parte dos indivíduos apresentava valores próximos ao ideal, o que pode ter sido uma variação do próprio método de avaliação da composição corporal. Observamos que 4 indivíduos apresentavam percentual de gordura adequado. Em relação ao IMM, os valores médios estavam dentro da normalidade (10,9 ± 0,6 kg/m2 ), sendo que apenas 4 atletas apresentavam pouca massa muscular. Conclusão: Quase todos os atletas apresentaram massa muscular dentro da normalidade, enquanto que a maioria apresentou valores acima do adequado de percentual de gordura. Estes resultados mostram que devem ser realizadas futuras intervenções nutricionais nestes atletas, focando principalmente na redução do percentual de gordura

Análise de longo prazo e obtenção do ano meteorológico típico para a usina fotovoltica flutuante de sobradinho

Para avaliar a geração de energia de centrais fotovoltaicas, é necessário analisar as variáveis ambientais associadas ao recurso solar na região de interesse durante vários anos. Neste trabalho, medições realizadas no Lago de Sobradinho de irradiância global inclinada, temperatura ambiente e velocidade do vento são utilizadas para corrigir séries de longo prazo dessas variáveis disponibilizadas por bases de dados históricas. Após obter as séries de longo prazo adaptadas ao local, um passo a passo é descrito e exemplificado para obtenção do ano meteorológico típico para a região de interesse. Em seguida, o ano meteorológico típico obtido é usado como entrada no software PVsyst previamente parametrizado de acordo com as características da Usina Fotovoltaica Flutuante (UFF) de Sobradinho para avaliar a geração de energia típica da usina. Os resultados mostram que a geração típica de energia da UFF - Sobradinho no ano inicial do projeto deve ser, em média, de 1797 MWh/ano.To assess the energy generation of photovoltaic plants, it is necessary to analyze the environmental variables associated with the solar resource in the region of interest for several years. In this work, measurements performed at Sobradinho's Lake of global irradiance on inclined plane, ambient temperature and wind speed are used to correct long-term series of these variables provided by historical databases. After obtaining the long-term series adapted to the location, a step by step is described and exemplified to obtain the typical meteorological year for the region of interest. Then, the typical meteorological year obtained is used as input in the PVsyst software previously parameterized according to the characteristics of the floating photovoltaic power (FPP) plant of Sobradinho to evaluate the typical power generation of the plant. The results show that the typical energy generation of Sobradinho FPP in the initial year of the project should be, on average, 1797 MWh/year.Asociación Argentina de Energías Renovables y Medio Ambiente (ASADES

Global pressures, specific responses: effects of nutrient enrichment in streams from different biomes

Fil: Artigas, Joan. Clermont Université. Université Blaise Pascal. Laboratoire Microorganismes: Génome et Environnement; FranceFil: García-Berthou, Emili. Institute of Aquatic Ecology. University of Girona. Girona; SpainFil: Bauer, Delia Elena. Instituto de Limnología Dr. Raúl A. Ringuelet (ILPLA). Facultad de Ciencias Naturales y Museo. Universidad Nacional de La Plata; ArgentinaFil: Castro, Maria I.. Department of Biology. National University of Colombia. Bogotá DC; ColombiaFil: Cochero, Joaquín. Instituto de Limnología Dr. Raúl A. Ringuelet (ILPLA). Facultad de Ciencias Naturales y Museo. Universidad Nacional de La Plata; ArgentinaFil: Colautti, Darío César. Instituto de Limnología Dr. Raúl A. Ringuelet (ILPLA). Facultad de Ciencias Naturales y Museo. Universidad Nacional de La Plata; ArgentinaFil: Cortelezzi, Agustina. Instituto de Limnología Dr. Raúl A. Ringuelet (ILPLA). Facultad de Ciencias Naturales y Museo. Universidad Nacional de La PlataFil: Donato, John C.. Department of Biology. National University of Colombia. Bogotá DC; ColombiaFil: Elosegi, Arturo. Faculty of Science and Technology. The University of the Basque Country. Bilbao; SpainFil: Feijoó, Claudia S.. INEDES. Department of Basic Sciences. National University of Luján. Luján; ArgentinaFil: Giorgi, Adonis. INEDES. Department of Basic Sciences. National University of Luján. Luján; ArgentinaFil: Gómez, Nora. Institute of Aquatic Ecology. University of Girona. Girona; SpainFil: Leggieri, Leonardo. Institute of Aquatic Ecology. University of Girona. Girona; SpainFil: Muñoz, Isabel. Department of Ecology. University of Barcelona. Barcelona; SpainFil: Rodrigues Capítulo, Alberto. Instituto de Limnología Dr. Raúl A. Ringuelet (ILPLA). Facultad de Ciencias Naturales y Museo. Universidad Nacional de La Plata; ArgentinaFil: Romaní, Anna M.. Institute of Aquatic Ecology. University of Girona. Girona; SpainFil: Sabater, Sergi. Catalan Institute for Water Research (ICRA). Scientific and Technological Park of the University of Girona. Girona; Spai

Estudo da marcha em idosos: resultados preliminares

The objectives of this pilot study, were the evaluation of, time-spatial, parameters of Brazilian elderly gait and to compare comfortable gait speed’s value with foreign reference data (from Oberg). Methods: Subjects were 15 healthy volunteers (8 men, 7 women) 60 to 79 years of age. The measurements were made in theirs building playground. Gait was timed over a 6 m distance; step length, stride width and foot angle were measured from footprints. Cadence was calculated from velocity and step length. Results: mean comfortable speed was ranged from 1,05 + 0,14 m/s for women in their sixties to 1,10 + 0,13 m/s for men in their seventies. Mean step length, stride width, cadence and foot angle were respectively 52,1 + 8,75 cm; 11,2 + 3,49 cm; 119,4 + 11,07 step/min and 13,5 + 8,53 degree for men and 46,6 + 8,08 cm; 6,75 + 7,07cm; 137,4 + 22,64 step/min and 7,5 + 5,1 degree for women. Conclusion: The lowest elderly’s gait speed obtained despite the sample’s small number in confronting with Oberg data, suggest the importance of comprehensive studies to supply the lack of normative gait data for the Brazilian population.Os objetivos foram avaliar parâmetros tempo-espaciais da marcha de idosos brasileiros e comparar o valor médio da velocidade confortável da marcha com um banco de dados estrangeiro (de Oberg) de parâmetros básicos da marcha. Metodo: Foram estudados 15 voluntários saudáveis (8 homens, 7 mulheres) dos 60 aos 79 anos de idade. As medidas foram realizadas no playgroung dos prédios onde residiam. A velocidade da marcha foi medida para uma distância de 6 m; o comprimento do passo, a largura da passada e o ângulo dos pés foram medidos a partir de impressões plantares. A cadência foi calculada a partir da velocidade da marcha e do comprimento do passo. Resultados: O valor médio da velocidade confortável da marcha variou de 1,05 + 0,14 m/s para mulheres da faixa etária de 60 anos a 1,10 + 0,13 m/s para homens da faixa etária de 70 anos. Os valores médios do comprimento do passo, da largura da passada, do ângulo dos pés e da cadência foram respectivamente 52,1 + 8,75 cm; 11,2 + 3,49 cm; 119,4 + 11,07 passos/min e 13,5 + 8,53 graus para os homens e 46,6 + 8,08 cm; 6,75 + 7,07cm; 137,4 + 22,64 passos/min e 7,5 + 5,1 graus para as mulheres. Conclusão: O menor valor da velocidade da marcha encontrado para os nossos idosos (apesar da casuística pequena), quando confrontado com os dados de Oberg, sugere a importância de estudos completos para suprir a falta de dados normativos de parâmetros da marcha para a população brasileira