Metal ions bound to the human milk immunoglobulin A: Metalloproteomic approach

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

Pluronics F-127/L-81 Binary Hydrogels as Drug-Delivery Systems: Influence of Physicochemical Aspects on Release Kinetics and Cytotoxicity

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

Análise eletroforética de proteínas recombinantes obtidas de Pichia pastoris

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.

Análise de metilação do gene FOXE1 em carcinoma de células escamosas em cães: padronização metodológica

It is believed that epigenetic mechanisms such as DNA methylation are important for the tumorigenesis and maintenance of the altered state of tumor cells. DNA methylation occurs by the addition of a methyl group to carbon 5 of cytosine, catalyzed by the enzyme DNA methyl-transferase, which can change the expression of a gene, including the tumor suppressor genes. In human squamous cell carcinoma, several features have shown the etiological role of genes in tumor development. Among them, FOXE1 gene (forkhead box E1 - thyroid transcription factor) is presented with an important role in susceptibility to disease. Similarly the FOXE1 methylation pattern could alter the expression of this gene in dogs and predisposed to tumor on. Therefore, this study aims to investigate in dogs, the validity of the strategy employed in humans to analyze the FOXE1 methylation status. DNA extraction from fresh frozen tumoral samples was performed by Wizard Genomic® DNA Purification Kit. The methylation status was determined by MSP-PCR (methylation-specific polymerase chain reaction), using 2.0 ng of DNA treated with sodium bisulphate. One hundred micrograms of bisulphite-modified DNA was amplified using primers specific for either methylated or unmethylated DNA (primers sequences are available at http://pathology2.jhu.edu/pancreas/primer.pdf). The analysis of fragments was loaded on to 7% polyacrylamide gels and silver nitrate staining. In this stage of technical approach, 60% were FOXE1 hypermethylated. In conclusion, it was observed that the standard technique for assessing the methylation pattern of gene FOXE1 in humans can be used for the same evaluation in dogs. The correlation of these molecular data with clinical and histopathological parameters may have diagnostic and prognostic value and still be used as a tumor marker for therapeutic decision and surgical approach

Determinação de Cd, Cu, Ni, Pb e Zn em águas marinhas oriundas da Bacia Potiguar utilizando a técnica de difusão em filmes finos por gradiente de concentração (DGT)

The DGT technique has been used to determine and better understand the dynamics and bioavailability of metals in the sea near to marine outfalls. In this work, the DGT was assessed for the determination of Cd, Pb, Cu, Zn and Ni in marine water samples from Potiguar Basin in its conventional aspect (binding agent, Chelex-100 and diffusive agent: Diffusive polyacrylamide gel (D) and restrictive (R)) and determination of Pb and Cd through its alternative aspect (binding agent: Saccharomyces cerevisiae immobilized in agarose gels and diffusive agent: 3MM Chromatography Paper). The deployment curves for long periods of immersion in the sample, showed a good linear correlation for the conventional aspect to Zn (D: R²=0.9586, R: R²=0.9444), Ni (D: R²=0.9789, R: R²=0.9286) and Cu (D: R²=0.764, R: R²=0.8143), and alternative to Pb (R²=0.9228) and Cd (R²=0.9673). The results of the organic and inorganic composition analysis in conventional aspect, showed that for every element there is no significant fraction of organic compounds in the sample. In addition, considering the alternative aspect, the comparison of obtained and expected masses suggests that some of the Pb is not labile and that the main retention mechanism of Pb for S. Cerevisiae occurs through ion exchange

Avaliação da membrana P81 como agente ligante da técnica de difusão em filmes finos por gradiente de concentração (DGT) para determinação de mercúrio em sistemas aquáticos

Worldwide environmental degradation is an undesirable byproduct resulting from the increasing demand for natural resources. Water sources are suffering intense contamination since they usually receive a huge amount of domestic and industrial effluents - which are mostly wasted without proper treatment - inserting a large number of pollutants in the environment, heavy metals included. Mercury holds great toxicological importance because, under some physicochemical conditions in a water environment, Hg (II) ion turns into methylated compounds stemming from this element, such as methylmercury CH3Hg, which is highly toxic for the aquatic community in which bioaccumulation occurs. Nowadays passive sampling techniques are being developed to enable the analytical procedures which are applied in environmental monitoring. Diffusive gradients in thin-films technique (DGT) has been proven an interesting tool for the determination of labile metal species due to its in situ application. The DGT technique consists of a piston-like device on which the following series of agents is disposed: a binding agent (conventionally Chelex 100 resin), a diffusive agent, usually a polyacrylamide gel, and a membrane filter. Nevertheless, the agents conventinally used for this technique don't usually show satisfactory results in mercury sampling. The main goal of this study was to evaluate the phosphate-treated cellulose membrane (Whatman P 81), an alternative material, as binding agent in the DGT to determine labile mercury fractions in aquatic systems. In this context, we conducted a study of the behavior of this material in relation with system variables, pH and ionic strength. Afterwards we performed immersions of the DGT devices in real and enriched samples and in situ aiming the determination of mercury

Concentrações séricas e peritoneal de proteínas de fase aguda em equinos submetidos à endotoxemia experimental, tratados ou não com lidocaína

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

Expressão heteróloga e modelagem de uma enzima lipolítica utilizando a abordagem metagenômica

Pós-graduação em Microbiologia Agropecuária - FCAV

Análises isoenzimáticas, moleculares e ultraestruturais em camarões dulcícolas do gênero Macrobrachium

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Abordagem metalômica quantitativa de mercúrio em peixes da região amazônica

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

Modificação superficial de nanopartículas de óxido de ferro por poli(ácido aspártico) reticulado

Pós-graduação em Química - IQ

Obtenção e estudo de polímeros supramoleculares derivados de ácidos carboxílicos-meglumina em fase aquosa

Pós-graduação em Química - IQ

Epidemiologia molecular de rotavírus em rebanhos bovinos no estado de São Paulo no período de 2006 a 2010

Pós-graduação em Medicina Veterinária - FCAV

DDRT-PCR approaches applied for preeminent results in the isolation of DETs from fish brain tissues

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

Eletroforese das proteínas séricas e urinárias de cães com erliquiose na fase aguda

Pós-graduação em Medicina Veterinária - FCAV