Metalloproteomic profile determination of muscle samples from nile Tilapia (Oreochromis niloticus) using AAS and ESI-MS/MS after 2D-PAGE separation

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

Eucalyptus urograndis stem proteome is responsive to short-term cold stress

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

Potential application of Ganoderma lucidum in solid state fermentation of primary sludge and wheat straw

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Mercury fractionation in dourada (Brachyplatystoma rousseauxii) of the Madeira River in Brazil using metalloproteomic strategies

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

Padronização de técnica manual para obtenção de plasma rico em plaquetas de bovino

To evaluate the biochemical profile and protein concentration of whey from milk samples of healthy Murrah primiparous and pluriparous buffaloes, 30 female buffaloes were analyzed during a complete lactation. The animals were divided into three groups: G1 = 10 primiparous buffaloes, G2 = 10 pluriparous buffaloes with 2-3 lactations and G3 = 10 pluriparous buffaloes with > 3 lactations. The lactation period was divided into: early stage (I: 1-3 months of lactation), intermediate stage (T: 4-6 months of lactation) and final stage (F: 7-9 months of lactation). Before milk sampling, physical examination of the mammary gland, strip cup test and California Mastitis Test (CMT) were performed. After mammary quarters asepsis, 20mL of milk were collected monthly from each mammary quarter, during a complete lactation, in sterilized plastic bottles without preservative, in order to perform microbiological isolation, biochemical profile and protein electrophoresis in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and 30mL of milk from each mammary quarter were collect, in sterilized plastic bottles containing preservative bronopol to perform the somatic cell count (SCC). A total of 1,042 milk samples were collected from the experimental groups during lactation, of which 923 samples showed negative reaction to CMT and negative microbiological isolation and were selected to biochemical profile analysis and protein electrophoresis in SDS-PAGE. There were influence of parity order and stage of lactation in biochemical profile and protein concentration of healthy Murrah buffaloes'whey. Primiparous buffaloes (G1) showed higher gamma-glutamyltransferase (GGT: 2,346 U/L), alkaline phosphatase (ALP: 181 U/L), phosphorus (P; 56.6mg/dL), potassium (K; 32.0mg/dL) and alpha-lactalbumin (458mg/dL). Buffaloes with 2-3 lactations (G2) showed higher SCC (70,700 cells/mL) and higher concentrations of total protein (1.55g/dL), albumin (100mg/dL), magnesium (Mg; 8.80mg/dL), chlorides (Cl; 176mg/dL), iron (Fe; 10.7 mu g/dL), sodium (Na; 178mMol/L) and lactoferrin (59.5mg/dL). Bufalloes with > 3 lactations (G3) showed higher concentrations of total calcium (Ca; 41.8mg/dL), ionized calcium (iCa; 2.92mMol/L), immunoglobulin A (IgA; 1.32mg/dL), serum albumin (99.1mg/dL), immunoglobulin G (IgG; 49.7mg/dL) and beta-lactoglobulin (1,068mg/dL). During lactation it was observed increase in SCC, GGT, ALP, total protein, albumin, P, Mg, Cl, Na, lactoferrin, serum albumin, IgG and alpha-lactalbumin, as well as decrease in concentrations of Ca, Fe, iCa, K, IgA and beta-lactoglobulin in buffaloes'whey. The results may be used as reference for buffaloes and to support diagnosis and prognosis of diseases common to lactation periods.

Community-associated methicillin-resistant Staphylococcus aureus in non-outbreak skin infections

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

Caracterização química dos géis produzidos pelas bactérias diazotróficas Rhizobium tropici e Mesorhizobium sp.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Polyphenol oxidase from sweet potato: Purification and properties

Polyphenol oxidase (PPO, EC 1.14.18.1) extracted from sweet potato root [Ipomoea batatas (L.) Lam.] was purified 189-fold by precipitation with ammonium sulfate and elution from columns of Sephadex G-25, DEAE-cellulose, and Sephadex G-100. Polyacrylamide gel electrophoresis of the purified preparation revealed that PPO was highly purified by the procedure adopted. The purified enzyme had an estimated molecular weight of 96 000 and Km values of 26, 8, 5, and 96 mM for 4-methylcatechol, chlorogenic acid, caffeic acid, and catechol, respectively. The optimum pH varies from about 4.0 to 6.5, depending on the substrate. PPO activity was inhibited by p-coumaric and cinnamic acids, sodium metabisulfite, dithioerythritol, ascorbic acid, L-lysine, D-phenylalanine, L-methionine, glycine, L-isoleucine, and L-glutamine. Heat inactivation between 60 and 80 °C was biphasic. Sucrose, (NH4)2SO4, NaCl, and KCl appeared to be protective agents of sweet potato PPO against thermal denaturation. © 1992 American Chemical Society.

Hemoglobin patterns in different populations of Synbranchus marmoratus Bloch, 1795 (Pisces, Synbranchidae)

1. 1. Total hemolysates of Synbranchus marmoratus Bloch, 1795 captured at four different sites in the State of São Paulo, Brazil, showed two different hemoglobin phenotypes when submitted to agar-starch gel electrophoresis on glass slides in basic buffer. 2. 2. Phenotype I was characterized by 3 hemoglobin bands. When the total hemolysate was submitted to cellulose acetate electrophoresis in basic buffer containing 6 M urea and β-mercaptoethanol, Phenotype I showed four globins of the α 1, α 2, β and γ types, with 11.9 ± 1.9 g% total hemoglobin, 45.3 ± 3.6% globular volume, and 26.8 ± 4.4% mean corpuscular hemoglobin concentration (MCHC). 3. 3. Phenotype II showed three groups of hemoglobins, with a total of up to 12 hemoglobin bands. When the total hemolysate was submitted to cellulose acetate electrophoresis in basic buffer containing 6 M urea and β-mercaptoethanol, phenotype II showed five types of globins, denoted types α 1, α 2, γ 1, γ 2 and β, having electrophoretic positions different from those of Phenotype I globins, with 18.1 ± 3.3% total hemoglobin, 47.9 ± 6.4% globular volume, and 37.8 ± 4.4% MCHC. 4. 4. The distribution of the specimens having the two hemoglobin phenotypes is associated with the different geomorphological provinces of the State of São Paulo, suggesting the existence of at least two populational groups of Synbranchus marmoratus. © 1986.

Determination of the mercury fraction linked to protein of muscle and liver tissue of Tucunaré (Cichla spp.) from the Amazon Region of Brazil

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

Clonal relatedness and resistance patterns of Salmonella Corvallis from poultry carcasses in a Brazilian slaughterhouse

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Characterization of the hemoglobins and globins of Synbranchus marmoratus Bloch, 1795 (Pisces, Synbranchidae)

1. 1. Total hemolysates of Synbranchus marmoratus Bloch, 1795, captured in Vitoriana, district of Botucatu, State of São Paulo, Brazil, were submitted to agar-starch gel electrophoresis on glass slides using 42 mM-Tris 1.7 mM EDTA-6.1 mM borate buffer, pH 8.8, for the gel and 10 mM borate-1.7 mM NaOH buffer, pH 8.6, for the cuvette. 2. 2. Three distinct hemoglobin bands were detected, with Hb I being of the cathodic type. 3. 3. Cellulose acetate electrophoresis in 800 mM Tris-2.1 mM EDTA buffer, pH 8.9, containing 6 M urea and 2.25 mM β-mercaptoethanol indicated the presence of four globin chains denoted α 1, α 2, β and γ. 4. 4. It is suggested that the probable tetrameric constitution of the hemoglobin of Synbranchus marmoratus Bloch, 1795 is Hb I (α 2 2γ 2), Hb II (α 2 1γ 2) and Hb III (α 2 1β 2). © 1986.

Proteinograma sérico de bezerros recém-nascidos alimentados com colostro de vacas com mastite

The aim of this study was to evaluate the serum protein concentration in newborns fed with colostrum derived from healthy cows (n = 10), cows with subclinical mastitis (n = 10) and cows with clinical mastitis (n = 10). 30 Holstein cows were assigned to their respective groups according to macroscopic examination of colostral secretion, somatic cell count, CMT and presence of bacteria in colostrum samples. Blood samples of the calves were collected immediately after birth, at 24 and 48 hours after ingestion of colostrum. The total protein was measured by the biuret method and the concentrations of immunoglobulin A (IgA), immunoglobulin G (IgG), transferrin, albumin and haptoglobin was determined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). No differences were obser- ved amongst groups in the concentrations of albumin, total protein and IgA. In animals from cows with subclinical and clinical mastitis haptoglobin concentrations were higher than those of healthy animals. The concentrations of IgG and transferrin were significantly lower in calves from cows with mastitis. We concluded that the ingestion of colostrum from infected and uninfected glands from cows with mastitis (GII e GIII) is unlikely to be an important contributor to the high rate of failure of passive transfer of immunoglobulins in calves.

Comparação entre dois métodos de detecção de DNA de papilomavírus humano em carcinoma epidermoide de lábio

Introdução: Recentemente o papilomavírus humano (HPV) tem sido associado à carcinogênese oral. A metodologia empregada na detecção do vírus é uma das maiores causas observadas da grande variabilidade nas taxas de detecção do HPV. Objetivo: Este estudo comparou a sensibilidade de detecção do DNA do HPV em casos de carcinoma epidermoide de lábio utilizando a amplificação do DNA viral por reação em cadeia da polimerase (PCR) ou nPCR. Material e método: Foram utilizadas 33 amostras provenientes de casos de carcinoma epidermoide de lábio. Para as extrações do DNA utilizou-se o sistema QIAamp DNA Mini Kit. Como controle interno utilizou-se o gene da b-globina. Das 33 amostras iniciais, 30 foram positivas para o gene b-globina, sendo utilizadas para detectar o DNA viral. Comparou-se a amplificação do DNA viral pelos métodos da PCR com os oligonucleotídeos MY09/MY11 e nPCR, empregando-se os pares de oligonucleotídeos iniciadores MY09/MY11 e, na segunda etapa, o par GP5+/GP6+. O controle positivo para a presença do DNA do HPV utilizado foi a linhagem de células HeLa e, como controle negativo, a mistura de amplificação sem DNA. A análise dos produtos de PCR e nPCR para HPV foi realizada por eletroforese em gel de poliacrilamida a 8%. Resultados: Utilizando-se o método da PCR, a amplificação do DNA do HPV foi constatada em dois casos. Com a nPCR foi verificada presença de DNA viral em 13 das 30 amostras. Conclusão: Com a utilização da nPCR, a detecção do HPV nos casos estudados aumentou mais de seis vezes.

Epidemiologia molecular e estudo dos fatores de virulência de Staphylococcus aureus resistentes à oxacilina isolados de feridas em pacientes atendidos em unidades básicas de saúde da cidade de Botucatu

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