Adult white leghorn hens hyperimmunised with Brazilian snake venoms of the genus Bothrops and/or Crotalus produced antibodies capable of recognising, combining with and neutralising the toxic and lethal components of the venoms. The antibodies were first detected by an enzyme-linked immunosorbent assay two weeks after starting the immunisation schedule, reached the highest titres by the third week and remained high for at least 24 weeks. These antibodies are transferred to the egg yolk from which they were isolated as enriched IgY preparations by a combination of methods using positive and negative precipitation with sodium sulphate and/or caprylic acid. The yolk-derived IgY preparations contained antibodies which blocked the phospholipase A2-dependent haemolytic activity of both venoms and the haemorrhagic activity of Bothrops venom, and neutralised the toxic lethal activities of the venoms with good efficacy. The median effective dose (ED50) of the IgY anti-Bothrops venom was 592.5 microliters/2LD50 and, 1.0 ml neutralised 0.0675 mg of venom. The ED50 of the IgY anti-Crotalus venom was 457.5 microliters/3LD50 and 1.0 ml neutralised 0.075 mg of venom.
ResumoInga striata Benth., popularmente conhecida como ingazeiro, é uma frutífera nativa das florestas tropicais e matas ciliares da Amazônia, Nordeste e Minas Gerais. É utilizada na recuperação de solos de áreas degradadas e na arborização urbana e rural. O objetivo do trabalho foi determinar os índices mais adequados para avaliação da maturidade e o ponto ideal de colheita das sementes de I. striata Benth. As árvores de I. striata estavam em floresta de brejo de altitude no Campus II da Universidade Federal da Paraíba (UFPB), em Areia-PB. As avaliações da maturação das sementes foram realizadas em seis épocas de colheitas de frutos após a antese (95, 110, 125, 140, 155 e 170 DAA) e em dois anos de observações consecutivos (duas safras). O delineamento utilizado foi o inteiramente casualizado seguindo um esquema fatorial 2 x 6 (dois períodos de observação e seis épocas de colheitas dos frutos). Em cada colheita foram avaliadas as dimensões de frutos e sementes, teor de água e massa seca das sementes, bem como a germinação e o vigor (primeira contagem de germinação, índice de velocidade de germinação, comprimento e massa seca da raiz primária e parte aérea). A maturidade fisiológica das sementes foi atingida aos 146 a 166 dias após a antese no primeiro ano e aos 155 dias no segundo ano, períodos em que apresentaram o menor teor de água, máximo acúmulo de massa seca e poder germinativo; O tamanho, teor de água, massa seca da semente, capacidade germinativa e ainda a massa seca da parte aérea da plântula, foram os parâmetros que melhor determinaram a maturação fisiológica das sementes de I. striata. Palavras-chave: Época de colheita, germinação, vigor, maturidade fisiológica AbstractInga striata Benth., Popularly known as ingazeiro, is a native fruit of tropical forests and riparian forests of the Amazon, Northeast and Minas Gerais. It is used in the reclamation of degraded areas and in rural and urban forestry. The objective of this study was to determine the most appropriate indices for assessing the maturity and ideal harvest seeds I. striata Benth. Trees I. striata were in swamp forest of altitude on Campus II of the Federal University of Paraíba (UFPB), Areia-PB. Assessments of seed maturation were performed in six seasons fruit harvests after anthesis (95, 110, 125, 140, 155 and 170 DAA) and two consecutive years of observations (two seasons). The experimental design was completely randomized following a factorial 2 x 6 (two periods of observation and six seasons crop of fruit). At each harvest were evaluated dimensions of fruits and seeds, water content and dry weight of seeds and their germination and vigor (first germination, speed of germination, length and mass of the primary root and shoot). The seed physiological maturity was reached at 146 to 166 days after flowering in the first year and 155 days in the second year, in which periods had the lowest water content, maximum dry matter accumulation and germination; The size, content water, dry mass of seed germination and the dry weight of the aerial p...
SUMMARYCoeliac disease (CD) is described as an autoimmune enteropathy associated with the presence of IgG and IgA antigliadin and antitransglutaminase autoantibodies. While of diagnostic significance, the role of these autoantibodies in the immunopathogenesis of CD is elucidated. An inappropriate T cell immune response to gluten is also involved in the pathogenesis of CD, as evidenced by autoantibody switching. The N-glycans released from serum IgG of CD patients and three groups of healthy controls, of differing age ranges, were analysed by NH2-high performance liquid chromatography (HPLC). The fucosylated biantennary N-glycans were the most abundant neutral oligosaccharides; in particular, the agalacto form (G0F) showed a mean value of 42% (s.d. ± 7·4), 30% (s.d. ± 5·9), 26% (s.d. ± 4·2) and 35% (s.d. ± 6·8) for CD patients, healthy children, healthy adults under 40 and healthy adults over 40 years old, respectively. The ratio of asialo agalacto fucosylated biantenna to asialo monogalacto fucosylated biantenna (G0F)/(G1F) for CD patients showed a significant increase compared to healthy children ( P < 0·0002), healthy adults under 40 ( P < 0·0002) and healthy adults over 40 years old ( P < 0·01). Hypogalactosylation was more pronounced for CD patients than for the patients with other autoimmune diseases such as rheumatoid arthritis or psoriatic arthritis.Keywords autoimmune disease coeliac disease immunoglobulin G oligosaccharides
-(Partial purification of trypsin/papain inhibitors from Hymenaea courbaril L. seeds and antibacterial effect of protein fractions). The crude extract and protein fractions of Hymenaea courbaril L. seeds were investigated for the presence of trypsin and papain inhibitors and antimicrobial activity against Vibrio parahaemolyticus, Staphylococcus aureus, and Escherichia coli. Protein fractions were obtained from the crude extract after precipitation with ammonium sulfate into three saturation ranges (0-30%, 30-60%, and 60-90%), called Hc030, Hc3060, and Hc6090, respectively. The crude extract and protein fractions inhibited trypsin and papain activity, but to different degrees. Antimicrobial activity was observed in Hc030 and Hc3060 fractions, but only against V. parahaemolyticus.The inhibitor isolated from the Hc3060 fraction was more effective in inhibiting trypsin (100% inhibition) than papain (54% inhibition), and showed an apparent molecular mass of 20 kDa. This study shows that H. courbaril seeds contain proteins with protease-inhibiting and antibacterial activity, indicating that this species is a source of bioactive compounds. Keywords: antimicrobial, cystatin, Fabaceae, protease RESUMO -(Purificação parcial de um inibidor de tripsina/papaína de sementes de Hymenaea courbaril L. e atividade antibacteriana de suas frações proteicas). O extrato bruto e frações proteicas de sementes de Hymenaea courbaril foram investigados para a presença de inibidores de tripsina e papaína e atividade antimicrobiana contra Vibrio parahaemolyticus, Staphylococcus aureus e Escherichia coli. As frações foram obtidas após precipitação do extrato bruto com sulfato de amônio em três faixas de saturação (0-30%, 30-60% e 60-90%) e denominadas Hc030, Hc3060 e Hc6090, respectivamente. O extrato bruto e as frações apresentaram diferente especificidade em inibir a atividade da tripsina e papaína. A atividade antimicrobiana foi observada nas frações Hc030 e Hc3060 e somente contra V. parahaemolyticus. O inibidor isolado a partir de Hc3060 foi mais eficiente na inibição da tripsina (100% de inibição) do que da papaína (54% de inibição), e mostrou uma massa molecular aparente de 20 kDa. Este estudo mostra que as sementes de H. courbaril possuem inibidores de proteases e proteínas com atividade antibacteriana, indicando que a espécie é uma fonte de compostos bioativos.
The aspartic proteinase gene of Mucor pusillus rennin expressed in Pichia pastoris was characterized in terms of structural and conformational stability induced by temperature. This enzyme is 12% glycosylated, with a similar specific activity to the native fungal enzyme. The secondary structure determined by CD is mainly due to beta-sheet structures with an important contribution of aromatic components. The calorimetric studies were carried out in the temperature range in which the enzyme is most stable. The enzyme undergoes an irreversible, highly scan-rate-dependent thermal denaturation under all the experimental conditions investigated. Between pH 3.0 and 7.0, only one endotherm characterized the thermal denaturation of enzyme. At pH 5.0, the most stable condition found, the denaturation can be fitted to the two-state irreversible model. Thus the kinetic constant and activation parameters of the denaturation process could be obtained. Upon reaching pH 7.5, the denaturation is characterized by two endotherms. This evidence indicates the complex tridimensional structure of this enzyme. Finally, taking into account the conservative tertiary structure of the aspartic proteinase family we comment on our results with reference to the crystallographic structure of M. pusillus pepsin [Newman, Watson, Roychowdhury, Badasso, Cleasby, Wood, Tickle and Blundell (1993) J. Mol. Biol. 221, 1295-1309].
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