476 resultados para purificação


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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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Pós-graduação em Engenharia Civil - FEIS

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Pós-graduação em Engenharia Civil - FEIS

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

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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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A esquistossomose é causada pelo Schistosoma mansoni e é considerada uma importante doença parasitária que afeta mais de 200 milhões de pessoas em países em desenvolvimento. Estima-se ainda que aproximadamente 600 milhões de pessoas vivam em áreas de risco e que o número de mortes por ano devido a esta parasitose chegue a 200.000. O tratamento existente hoje é eficiente, mas não previne contra re-infecção e contra as formas jovens dos parasitas. O desenvolvimento de uma vacina utilizando antígenos do parasita produzidos de modo recombinantes é o anseio para o controle da esquistossomose. A fosfodiesterase-5 (SmNPP-5a) é uma enzima presente na interface parasita-hospedeiro e acessível ao sistema imune, sendo portanto considerada um potencial candidato vacinal, sua avaliação se faz necessária. A obtenção de uma SmNPP-5a recombinante enzimaticamente ativa é um passo fundamental no processo de avaliação da sua capacidade protetora e determinação da sua função fisiológica para o parasita. Utilizando o sistema de expressão baseados em leveduras metilotróficas, Pichia pastoris, a proteína recombinante pode ser expressa com as modificações pós traducionais necessárias para ter sua estrutura original mantida, característica fundamental para se obter uma proteína enzimáticamente ativa. Para clonagem e expressão da proteína recombinante utilizamos o vetor pPICZαA, pois este possui um sistema de expressão baseado no promotor álcool oxidase (AOX1 e AOX2). Ele possui ainda, a capacidade de adicionar um peptídeo sinal e uma cauda de histidina, com objetivo de secretar a proteína expressa e facilitar sua purificação, respectivamente. A triagem e confirmação dos clones positivos foram feitas por PCR. A análise das amostras de sobrenadantes coletadas com 24, 48 e 72h de estímulo com metanol foi feita por SDS-PAGE e Western blot para verificação da expressão... (Resumo completo, clicar acesso eletrônico abaixo)

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The expansion of the energy matrix is a matter of great importance since the recent problems facing the country in this sector, such as rationing mobilized in 2001. Moreover, there is also concern with global sustainable development. Brazil is producing about 64 million cubic meters of biogas per day, but only a small percentage of this total is harnessed as an energy resource. Compounders are considered potential, for example, landfill energy (LE) and sewage treatment plants (STP), which can be adapted to produce biogas and market them. The work focuses its studies on biogas produced by these small and medium compounders. Are proposed and studied the following alternatives for the utilization of biogas energy: generation and sale of electricity through the installation of a small power station within the landfill, and purification and marketing of gas, whose price will be a consideration on the price of natural gas, taking into account the percentage of methane present in the biogas. Trade ensures that gas plus a destination ecologically appropriate to it, taking advantage of its energy

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Human development and population growth during the twentieth century increased the water demand, tripling its consumption between 1950 and 1990. As the water streams were polluted; and as water is the source of minerals and also regulates vital functions, it becomes the vehicle of transmission and consequently spreads many diseases. Probably, the industries are the major responsible for this pollution when they dump untreated effluents to water streams, saturating the already insufficient net of sanitation facilities polluting water and soil. An effective treatment has been established with low cost in Europe and the United States, through constructed systems on wetlands Constructed Wetland Systems - CWSs, gradually used in other countries in the last three decades. Lately, we observe a continuous growth in Brazilian poultry business, and poultry industry showed greatest dynamism in the country, following the global market. Pondering this information and the efficiency of such treatment, this work aimed to study prototypes, in a laboratory scale, simulating ascending and descending types of CWSs, vegetated with aquatic macrophytes Eichhornia crassipes and the uses of aggregates and soil, to treat industrial wastewater from slaughterhouses and aviary. We conducted the initial characterization of the effluent to have an idea of its constituents and to scale the system and the continuous flow. Furthermore, we characterized the soil to be used in this system. The collects are periodically made in the refrigeration industry FRICOCK FRIGORIFICAÇÃO AVICULTURA INDÚSTRIA E COMÉRCIO LTDA. for local treatment simulation. The effluent that was treated with 12 prototypes of CWSs are analyzed with some frequency. The results of these reviews were compared to the effluent coming from the industry... (Complete abstract click electronic access below)

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A number of methods and products have been developed in order to eliminate or reduce the negative effects that hydrocarbons cause to the environment, including hydrophobic expanded vermiculite, used in oil residue filtering systems at gas stations. However, upon adsorbing organic compounds, the vermiculite is no longer used and is sent to landfills. The aim of the present study was to wash granular and powdered vermiculite containing oil lubricant in its pores with distilled water and solutions of 0.1% SDS surfactant and rhamnolipids, with the aim of removing the lubricant and the possibility of reusing the mineral. The greatest amount of lubricant removal was obtained through washing with 0.1% SDS and both granulometric forms. This may be associated to the industrial purification received by the surfactant. However, the biosurfactant is ecologically more viable due to its low toxicity and ease of degradability. In the readsorption tests, greatest adsorption was obtained with the granular vermiculite washed in SDS solution. In order to enable the reuse of the mineral, further tests are needed to enhance desorption/adsorption efficiency.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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A glicerol quinase é uma proteína tetramérica que cataliza a fosforilação do glicerol a glicerol-3-fosfato, composta por quatro subunidades - sua análise eletroforética fornecerá apenas uma banda se for constatada a sua pureza num gel não desnaturante - e sua reação com o glicerol é dependente de magnésio e de ATP. A eletroforese de proteínas utiliza como suporte um gel de poliacrilamida, que é formado pela polimerização de monômeros de acrilamida ao longo da sua cadeia e ligações cruzadas de cadeias pela inclusão de um co-monômero bifuncional apropriado, usualmente a N,N’ – metileno-bis-acrilamida ou apenas Bis . A eletroforese de proteínas mais comum é a que utiliza SDS para um gel de poliacrilamida desnaturante. O SDS é um sal chamado Dodecil Sulfato de Sódio que tem por característica se ligar a cadeia proteína nos resíduos de aminoácidos apolares, deixando com a carga negativa, sendo assim, toda proteína fica com carga total negativa, migrando para o anodo na eletroforese. As técnicas de purificação utilizadas foram ultrafiltração e precipitação com ácido tricloro acético, etanol gelado e sulfato de amônio. A dupla precipitação com etanol resultou na recuperação de maior quantidade de proteínas. A coloração do gel com prata foi mais sensível do que Comassie blue. O gel de eletroforese mostrou quatro bandas, correspondentes às quatro subunidades da glicerol quinase quando revelados com prata em gel de SDS-PAGE.

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The volume of liquid effluent generated in cattle slaughterhouses is quite high and cannot be released untreated in water bodies due to its high pollution load of predominantly organic origin. To minimize the environmental impacts of its industrial wastewater and meet the local environmental legislation, abattoirs shall make the treatment of these effluents. The present work aims to develop the study of a reactor by sequential batch pilot scale, in order to optimize their performance in treating wastewater from a cattle slaughterhouse. The treatment system used was developed and installed in the Laboratory of Wastewater Treatment, in Faculty of Science and Technology UNESP, Presidente Prudente campus. The procedure used followed the operation of sequential batch reactors, in which all processes and treatment operations occurring sequentially in a single unit, by establishing specific operating cycles, which comprise the following separated phases: aerobic reaction, anoxic reaction, sedimentation and emptying. Aiming to improve the quality of treatment was planned the addition of coagulant Poly Aluminum Chloride (PAC) in the reactor, by determining their optimal dosage by Jar-test trials. Were prepared four steps with specific operating cycles: step one or acclimatization (10 hour of aeration, one hour and 30 minutes of sedimentation and 30 minutes for exchanging the effluent); step 2 (6 hours of aeration or aerobic phase, 4 hours and 45 minutes of stirring or anoxic phase and 1 hour and 15 minutes for sedimentation and exchange effluent); step 3 (2 hours and 30 minutes of aeration, 8 hours and 15 minutes of stirring and 1 hour and 15 minutes for sedimentation and exchange) and step 4 (2 hours of aeration, 8 hours and 45 minutes of stirring and 1 hour and 15 minutes for sedimentation and exchange)... (Complete abstract click electronic access below)