Vicilina de sementes da leguminosa selvagem Anadenanthera macrocarpa (AmV): purificação, caracterização, efeito deletério e mecanismo de ação para bruquídeo callosobruchus maculatus

Grains and legume seeds are foods that form the basis of the diets of many cultures around the world, winch contritbute to the daily nutrient requirements of humans. Vicilins (7S globulin) are storage proteins found in legume seeds, and may have an additional function constitutive defense of the embryo against pests and pathogens. In this work the vicilin from Anadenanthera macrocarpa - AmV (red-angico), was purified and partially characterized, its effect on development and larval survival and adult emergence of Callosobruchus maculatus was evaluated by determination of LD50, WD50 and ED50 in system bioassay. Purification of vicilin was initiated by the chitin affinity chromatography and then gel filtration (Superdex 75 Tricorn 10x300 mm) FPLC system followed by reverse phase chromatography (C8 phenomenex) on HPLC system. Bioassays WD50 and LD50 for larvae were 0.32% and 0.33% (w:w) respectively, since the ED50 for adults was 0.096%. The probable mechanism of action was evaluated by testing digestibility of AmV in vitro, and observed for the involvement of two fragments vicilins immunoreactive against polyclonal Anti-vicilin from Erythrina velutina (Anti-EvV) about of 22 and 13 kDa chitin binding. The AmV in its native form has been recognized by the anti-EvV, indicating that there is a conserved region in the vicilin and is probably corresponding to the chitin binding domains. These results point to a new vicilin chitin binding that can subsequently be used as a possible biopesticide protein source, in order to control insect pest C. maculatus and confirm literature findings that demonstrate vicilin in the presence of different kinds of ligands to conserved regions chitin not yet characterized

Estudo eritroleucométrico e proteinograma sérico do sangue do cordão umbilical e jugular de eqüinos ao nascimento e de suas respectivas mães

Colheram-se amostras de sangue do cordão umbilical (SCU) e do sangue circulante de cinco eqüinos neonatos, imediatamente após o nascimento, e o sangue da própria mãe, utilizando-se um sistema a vácuo. O material foi submetido à contagem global de hemácias e leucócitos e à determinação do volume globular e da concentração de hemoglobina; à contagem diferencial de leucócitos em esfregaços sangüíneos; e ao cálculo dos índices eritrocitométricos. Foram realizadas a dosagem de proteínas séricas totais e a eletroforese das proteínas séricas em gel de agarose. Não houve diferenças significativas entre os parâmetros do SCU e do sangue da jugular dos potros. No SCU dos potros observaram-se valores mais elevados para contagem global de hemácias (9,75x10(6)/µl), dosagem de hemoglobina (14,65g/dl) e concentração de hemoglobina corpuscular média (37,23g/dl); e valores menores para volume corpuscular médio (40,50fl), proteína total (4,37g/dl), alfa-globulinas (0,65g/dl), beta-globulinas (1,10g/dl), gama-globulinas (0g/dl) e contagens global (5,40 x 10³/µl) e diferencial de leucócitos, exceto contagem de neutrófilos bastonetes e monócitos, quando comparados com os valores obtidos no sangue de suas mães.

Proteinograma sérico de bovinos da raça Curraleir

Estudou-se o perfil eletroforético das proteínas séricas de bovinos sadios da raça Curraleiro por meio da técnica de eletroforese em gel de acrilamida contendo dodecil sulfato de sódio. Utilizaram-se amostras de soro sanguíneo de 228 bovinos da raça Curraleiro, 51 machos e 177 fêmeas, com idades entre sete meses e 12 anos, pertencentes a dois rebanhos localizados nos Estados de Goiás e Tocantins. Foram quantificadas proteína total e concentração plasmática de fibrinogênio. Verificaram-se variações nas concentrações das diferentes frações proteicas. Foram detectadas 26 proteínas e identificadas 10 delas. A ceruloplasmina esteve ausente em 78,1% dos indivíduos, e a α-antitripsina não foi detectada em nenhum animal. Proteína total, globulina, IgA, IgG e fibrinogênio aumentaram com a idade e houve correlação positiva entre os níveis séricos de haptoglobina e α1-glicoproteína ácida.

Hematological, biochemical and anatomopathological aspects of the experimental infection with Trypanosoma evansi in dogs

O presente estudo acompanhou durante 82 dias o curso da infecção experimental com T. evansi em quatro cães, realizando a avaliação dos achados hematológicos, bioquímicos e anatomopatológicos. Os animais infectados mostraram declínio acentuado na contagem de hemácias, hematócrito e teor de hemoglobina, permanecendo anêmicos a partir da terceira semana de infecção até o final do período experimental. Leucopenia com neutropenia foram observadas entre a segunda e a quinta semanas após a infecção. Os cães inoculados desenvolveram hiperproteinemia, sendo constatada diminuição na relação albumina:globulina. As atividades séricas de alamina aminotransferase e aspartato aminotransferase aumentaram significativamente nos cães infectados em relação aos animais controle. O exame histopatológico revelou hiperplasia linfóide no baço e linfonodos e infiltrado mononuclear periportal e esteatose de padrão centrolobular no fígado de todos os cães infectados. Intenso infiltrado mononuclear foi observado no miocárdio de três cães e acúmulos de células mononucleares junto às meninges foram evidenciados em dois animais infectados.

Características físico-químicas e citológicas do líquido sinovial da bainha tendínea digital de eqüinos

Foram estudadas as características físico-químicas e citológicas do líquido sinovial da bainha tendínea digital de nove eqüinos hígidos. Verificou-se que o líquido é viscoso, amarelo claro, límpido, livre de partículas e que não coagula à temperatura ambiente. Sua concentração média de ácido hialurônico foi 60,20mg/dl, a taxa de glicose, similar à plasmática e sua concentração protéica não ultrapassou 1,74g/dl, com relação média albumina:globulina de 0,94. O número médio de células nucleadas foi de 313 células/µl, com predominância de grandes células mononucleares e linfócitos. Houve correlação significativa (r = - 0,649, P<0,01) entre o aumento da concentração de ácido hialurônico e a diminuição percentual de linfócitos. As mensurações das características pesquisadas no líquido sinovial da bainha tendínea digital de eqüinos são de execução simples e passíveis de implantação na rotina de atendimentos clínico-cirúrgicos.

Dried yeast (Saccharomyces cerevisae) as a protein source for horses

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Sementes de feijão submetidas a ciclos e períodos de hidratação-secagem

A técnica de hidratação-secagem pode ocasionar alterações fisiológicas e bioquímicas em sementes de feijão e afetar sua qualidade fisiológica. Sementes do cultivar Carioca foram submetidas a tratamentos de hidratação-secagem por 6, 12 e 24 horas com três ciclos de hidratação, com o objetivo de avaliar os efeitos causados por períodos e ciclos de hidratação-secagem na qualidade fisiológica das sementes. O delineamento experimental utilizado foi inteiramente casualizado, com quatro repetições, analisado estatisticamente em esquema fatorial 3 x 3 com uma testemunha absoluta. Para a primeira contagem da germinação, os tratamentos de períodos combinados com ciclos de hidratação-secagem, apresentaram superioridade em relação ao tratamento testemunha. Na avaliação de grupos de proteínas, as concentrações de globulinas e prolaminas nas sementes submetidas aos tratamentos, foram significativamene superiores a testemunha. Os cátions (Ca, Mg e K) lixiviados para solução de embebição das sementes tiveram comportamento semelhante à condutividade elétrica das mesmas.

Enzymatic Hydrolysis of Sweet Lupin, Chickpea, and Lentil 11S Globulins Decreases their Antigenic Activity

We investigated the effects of treatments with the enzymes pepsin and trypsin on the in vitro immunological reactivity of the major globulins found in the seeds of sweet lupin, chickpea, and lentil. Polyclonal major globulin-specific antiserum was obtained by immunization of rabbits with a solution of the 11 S globulin of each legume. The globulins were hydrolyzed with pepsin and trypsin for 1, 5, 15, and 30 min. The native globulins and their hydrolysates were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting to identify the polypeptide bands with antigenic activity, and the hypoantigenicity of the hydrolysates was analyzed by enzyme-linked immunosorbent assay. Our results show that enzymatic treatment of the major storage protein (11 S globulin) of sweet lupin, chickpea, and lentil with pepsin or trypsin lead to the formation of large amounts of short peptides and free amino acids that do not allow antibody binding, resulting in a weakened immunoreactivity.

Nutritional Responses of Rats to Diets Based on Chickpea (Cicer arietinum L.) Seed Meal or Its Protein Fractions

The aim of this study was to isolate the protein fractions from chickpea, var. IAC-Marrocos, as well as to evaluate its in vivo nutritional protein quality. Among the proteins, albumins showed better nutritional value in the in vivo assays and amino acid contents, despite their higher trypsin inhibitor contents. Trypsin inhibitors were found to be heat labile in all samples, but the digestibility results for unheated and heated flour and albumins suggest that their contents are not very decisive. The PER values for casein (not supplemented) were very similar to those of heated flour and unheated or heated albumin and total globulins. The albumin and glutelin fractions showed the best results for PDCAAS, however, lower than those of casein. Despite the high digestibility of the globulin the very low essential amino acid content lowered its PDCAAS, and it had the lowest values.

Isolation, solubility and in vitro hydrolysis of chickpea vicilin-like protein

The chickpea vicilin-like globulin was isolated and chromatographed on Sepharose CL-6B and Sephacryl S-300. The native globulin with a molecular weight of 140 kDa was resolved in Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in seven polypeptide bands in the range of 12.4-67 kDa. The solubility profile of the protein in water and NaCl solutions was typical of a legume globulin. The purified vicilin-like globulin, native and heated, was hydrolyzed by pepsin, trypsin and chymotrypsin. The hydrolysis patterns indicated that the native vicilin-like protein was only partially degraded by the enzymes in comparison with casein. Heating increased its susceptibility to hydrolysis relative to the native form, for all the enzymes. However, the results obtained by the pH-drop method revealed that the in vitro digestibility of the vicilin-like protein was not altered by heating, while 11 S-like and total globulins suffered a small increase, indicating that the structural characteristics of storage globulins may be important factors limiting the protein digestion. (c) 2007 Swiss Society of Food Science and Technology. Published by Elsevier Ltd. All rights reserved.

Partial characterization of proteins from baru (Dipteryx alata Vog) seeds

BACKGROUND: Baru (Dipteryx alata Vog.) is a fruit distributed throughout the Brazilian savanna and contains a seed with a high protein content, whose properties have been rarely explored. The purpose of this study was to characterize this protein, especially by isolation and quantifying its fractions and measuring some of its molecular properties.RESULTS: Baru seeds contain 244 g kg(-1) protein on a dry weight basis. Solubility profiles showed a preponderance of globulins. This fraction dominated the seed composition, with 61.7 wt% of the total soluble proteins. Albumins and glutelins accounted for 14 and 3.3 wt%, respectively. SDS-PAGE resolution of albumin and globulin showed main bands with molecular weights of 84 kDa and 64,66 and 73 kDa, respectively. The total protein of the flour and the globulin showed values of in vitro digestibility of 85.59% and 90.54%, relative to casein. Total globulin produced only one chromatographic peak, both on Sepharose CL-6B gel filtration and on DEAE-cellulose ion-exchange columns, eluted at a concentration of 0.12 mol L(-1) NaCl.CONCLUSION: The baru seed had high protein content with large quantities of storage proteins. The chromatographic and solubility profiles indicate the predominance of a fraction with characteristics of a legumin-type protein. (C) 2011 Society of Chemical Industry

Soybean glycinin improves HDL-C and suppresses the effects of rosuvastatin on hypercholesterolemic rats

Background: This study was an investigation of the effects of ingesting a daily dose of isolated glycinin soy protein (11S globulin), in association with rosuvastatin, on the control of hypercholesterolemia in experimental animals.Methods: Male Wistar rats were kept in individual cages under appropriate controlled conditions of temperature, light and humidity. The animals were divided into five groups (n = 9): 1) standard (STD): fed on casein as protein source; 2) hypercholesterolemic (HC): STD plus 1% cholesterol and 0.5% cholic acid; 3) HC+11S: hypercholesterolemic + glycinin (300 mg/kg/day); 4) HC+ROS: hypercholesterolemic + rosuvastatin (10 mg/kg/day); 5) HC+11S+ROS: HC diet, the 11S protein and the drug in the doses given in (3) and (4). The protein and the drug were administered by gavage for 28 days. The results indicated that the addition of 1% cholesterol and 0.5% cholic acid induced hypercholesterolemia in the animals without interfering with their weight gain.Results: A single daily dose of glycinin contributed an additional 2.8% of dietary protein intake and demonstrated its functional role, particularly in raising HDL-C, decreasing triglycerides in the liver and improving the atherogenic index in animals exposed to a hypercholesterolemic diet.Conclusion: Most of the beneficial effects of the isolated treatments disappeared when the drug (rosuvastatin) and the protein (glycinin) were taken simultaneously. The association was shown not to interact additively, as noted in the plasma levels of total cholesterol and non-HDL cholesterol, and in the significant increase of cholesterol in the liver. Studies are in progress to identify the effects of peptides derived from the 11S globulin and their role in cholesterol metabolism.

Caracterização e hidrólise in vitro da globulina principal de grão-de-bico (Cicer arietinum L.), var. IAC-Marrocos

No presente estudo procedeu-se ao isolamento e caracterização da fração globulina majoritária (11 S) de grão-de-bico, var. IAC-Marrocos. A globulina majoritária extraída foi isolada por cromatografia de filtração em gel e de troca-iônica mostrando apenas uma banda de proteína na eletroforese em gel de poliacrilamida. A globulina majoritária, após passagem em coluna de Sephadex, revelou duas bandas protéicas de 55 e 52,5kDa e três bandas menores em gel de poliacrilamida dodecilsulfato de sódio. Na presença de 2-mercaptoetanol 6 polipeptídios na faixa de 18 a 42kDa foram revelados na eletroforese. A globulina isolada foi submetida à ação da tripsina e quimotripsina onde a forma nativa mostrou-se resistente à ação enzimática enquanto o aquecimento (96 e 121°C/15min) não foi suficiente para aumentar a susceptibilidade à hidrólise, significativamente. Adição de NaCl 0,3M levou a um aumento da estabilidade estrutural com menor susceptibilidade à digestão proteolítica, fato em parte perdido com o aquecimento. As hidrólises foram acompanhadas por eletroforese em gel de poliacrilamida dodecilsulfato de sódio.

Composição química e nutrientes em sementes das espécies florestais Pente de Macaco, Flor de Paca, Itaúba, Jatobá e Murici Manso

A considerable number of tree species is the flora in some cases, their fruits and seeds turn out to be good sources of nutrientes. The study aimed to verify the chemical composition of nutrientes in seeds of forest species Pente de Macaco, Flor de Paca, Ita ba, Jatoba and Murici Manso. of forest species of Monkey Comb, Flor de Paca, Ita ba, Jatoba and Murici Manso. The experimental design was randomized with five treatments. Analyses were carried out: moisture content, starch content (mg. g (1)), carbohydrates (mg. g (1)), and proteins (albumin, globulin, prolamin, glutelin) (mg. g (1)) and nutrients (N, P, K, Ca, Mg and S). The results showed that the seeds of Flor de Paca had the highest content of starch and protein of Jatoba is a significant source of carbohydrates and the highest levels of phosphorus, calcium and sulfur. For contents of nitrogen Ita ba showed the highest levels and also obtained satisfactory results for calcium and species Murici Manso had the highest levels of potassium and magnesium.

Influência do molibdênio e do cálcio aplicados via semente nas frações protéicas de amendoim cv.iac 886

Chemical composition of seed displays, in general, the same compounds found in other parts of the plant, and the environment where they grow plants, fertilizer and many other factors are able to change this constitution, increasing or decreasing the amount of certain components. The study aimed to determine the effect of application by seed doses of calcium and molybdenum on protein content of peanut seeds cv.IAC 886. The experimental design was randomized blocks in a factorial design with four replicates per treatment, which are constituted by the combination of molybdenum doses (0, 50, 100 and 150 g ha(-1)) and calcium by seeds (0, 1000, 2000 and 3000 mg L-1). The peanut harvest was done manually. Seeds were removed from the pod manually and individually for each treatment and were taken to the Laboratorio de Genetica de Populacoes e Silvicultura, do Departamento de Fitotecnia, Tecnologia de Alimentos e Socio-Economia, da Faculdade de Engenharia de Ilha Solteira da Universidade Estadual Paulista, where we determined the protein (albumin-Alb, prolamin-PRO, glutelin-GLU and globulin-GLO, mg g(-1). Regardless of the doses used the albumin protein fraction showed the highest in the peanut seeds. The addition of molybdenum resulted in increased seed prolamin content in the peanut seeds. The combination of calcium and molybdenum applied to seeds resulted in increased levels of albumin, globulin and glutelin in the peanut seeds.