938 resultados para Affinity purification


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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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O texto apresenta dois conjuntos de informações de interesse histórico para a Psicologia: sobre o contexto no qual foi regulamentada a profi ssão de psicólogo e sobre a formação em nível de pós-graduação. Tais informações são utilizadas para discutir a dificuldade de promover articulação de fato entre a formação em nível de graduação e a pós-graduação. Trata-se de dificuldade especialmente curiosa, pois desde a fase inicial da organização da pós-graduação brasileira, a necessidade de considerar sua integração com a formação em nível de graduação foi sempre ressaltada. Ainda assim, persistem dificuldades relativas à essa integração. Algumas proposições de atividades que podem ensejar articulações entre os diferentes níveis de formação são apresentadas para o debate, tanto na esfera do ensino e da orientação como no âmbito da pesquisa, havendo menção, também, a atividades correlatas que podem servir ao mesmo objetivo.

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The properties of a previously unknown enzyme, denominated cyclodextrin glycosyltransferase, produced from Bacillus lehensis, were evaluated using affinity chromatography for protein purification. Enzyme characteristics (optimum pH and temperature; pH and temperature stability), the influence of substances on the enzyme activity, enzyme kinetics, and cyclodextrin production were analysed. Cyclodextrin glycosyltransferase was purified up to 320.74-fold by affinity chromatography using beta-cyclodextrin as the binder and it exhibited 8.71% activity recovery. This enzyme is a monomer with a molecular weight of 81.27 kDa, as estimated by SDS-PAGE. Optimum temperature and pH for cydodextrin glycosyltransferase were 55 degrees C and 8.0, respectively. The Michaelis-Menten constant was 8.62 g/l during maximum velocity of 0.858 g/l.h.

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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INTRODUCTION: We aimed to evaluate the effects of musical auditory stimulation on cardiac autonomic regulation in subjects who enjoy and who do not enjoy the music. METHOD: The study was performed in young women (18-27 years old) divided in two groups (1) volunteers who enjoyed the music and (2) volunteers who did not enjoy the music. Linear indices of heart rate variability were analyzed in the time domain. The subjects were exposed to a musical piece (Pachelbel: Canon in D Major) during 10 minutes. Heart rate variability was analyzed at rest with no music and during musical auditory stimulation. RESULTS: In the group that enjoyed the music the standard deviation of normal-to-normal R-R intervals (SDNN) was significantly reduced during exposure to musical auditory stimulation. We found no significant changes for the other linear indices. The group composed of women who did not enjoy the music did not present significant cardiac autonomic responses during exposure to musical auditory stimulation. CONCLUSION: Women who enjoyed the music presented a significant cardiac autonomic response consisting of a reduction in heart rate variability induced by the musical auditory stimulation. Those who did not enjoy the musical piece presented no such response.

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An extracellular pectin lyase secreted by Fusarium decemcellulare MTCC 2079 under solid state fermentation condition has been purified to electrophoretic homogeniety by using ammonium sulfate fractionation, carboxymethyl cellulose and gel filtration (Sephadex G-100) column chromatographies. The purified enzyme showed single protein band corresponding to molecular mass 45 +/- 01 kDa on sodium dodecyl sulfate polyacrylamide gel electrophoresis. The enzyme had maximum activity at pH 9.0 and showed maximum stability in the pH range of 9.0-12.0. The optimum temperature of the purified enzyme was 50 degrees C and it showed maximum stability upto 40 degrees C. The energy of activation for the thermal denaturation (Ea) was 59.06 kJ mol(-1) K-1. The K-m and k(cat) values using citrus pectin as the substrate were 0.125mgml(-1) and 72.9 s(-1) in 100mM sodium carbonate buffer pH 9.0 at 50 degrees C. The biophysical studies on pectin lyase showed that its secondary structure belongs to alpha+beta class of protein with comparatively less of beta-sheets. Purified pectin lyase showed efficient retting of Crotolaria juncea fibers.

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Mesenchymal stem cells (MSCs) are a heterogeneous population of cells that proliferate in vitro as plastic-adherent cells, have fibroblast-like morphology and can differentiate into bone, cartilage and fat cells. Therapeutic potential of MSCs have been studied in experimental models, such as rabbit, in Laboratory of Cell Engineering of Botucatu. However, no specific markers have been reported for expanded rabbit MSCs, which hampers the isolation of pure MSC populations by immunophenotypic characterization. Thus, the objective of this study was to produce monoclonal antibodies (mAbs) to rabbit MSCs. MSCs derived from rabbit bone marrow (BM) were isolated, cultured, expanded ex vivo, and immunized into three BALB/c mices, and spleen cells subsequently harvested were used to generate hibridoma cell lines secreting antibodies against MSCs. Hybridoma cells were screened by flow cytometry and antibody-producing cells were subjected to subsequent rounds of retests. MSC1-160 obtained the best positivity for IgG expression and was cloned by limiting dilutions and micromanipulation. Ascitic fluid from ten best clones was purified by affinity chromatography in Protein A-sepharose CL-4B column and purification control was performed by electrophoresis in agarose gels. The purified IgG were tested against rabbit MSCs, obtaining high positivity by flow Cytometry. In conclusion, we developed 10 mAbs, MSC1-160 A20, A30, A41, A47, A55, A60, A63, A69, A81, and A82, that recognize rabbit MSC cell surface antigens showing potential for immunophenotypic characterization of rabbit MSC cell lines

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Pós-graduação em Agronomia (Genética e Melhoramento de Plantas) - FCAV

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