Heparin-binding proteins of seminal plasma in Nellore bulls Proteínas ligadoras à heparina do plasma seminal em touros Nelore

The aim of this study was to identify heparin-binding proteins (HBPs) in seminal plasma of Nellore (Bos taurus indicus) bulls. Bulls (n=4), 30-36 months old, 500-550kg with satisfactory seminal quality were selected. After the centrifugation, samples of the seminal plasma were pooled and the HBPs were isolated by heparin-affinity chromatography. The recovered HBPs fractions were pooled. One-dimensional sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDSPAGE) 12.5% was performed in vertical minigels. Eight bands with molecular weights ranging from 15 to 63kDa were observed. Two proteins were identified (22 and 25kDa), similar to those previously described in Bos taurus taurus bulls. Other bands identified in this study (39, 53, 58 and 63kDa) have not been previously observed and possibly they are specific to Nellore semen.O objetivo deste estudo foi identificar proteínas ligadoras à heparina no plasma seminal de touros Nelore (Bos taurus indicus). Para tanto, foram selecionados quatro touros entre 30 e 36 meses de idade e peso aproximado de 500-550kg. Após centrifugação, amostras do plasma seminal foram misturadas e as proteínas ligadoras à heparina foram isoladas por meio da cromatografia por afinidade. As frações após a eluição foram agrupadas para caracterização das bandas protéicas (SDSPAGE, 12,5%). Foram identificadas oito bandas protéicas variando entre 15 e 63kDa. Duas proteínas com 22 e 25kDa foram similares às descritas em touros Bos taurus taurus. Outras proteínas identificadas com 39, 53, 58 e 63kDa ainda não foram descritas e possivelmente sejam específicas para Bos taurus indicus

Heparin-binding proteins of seminal plasma in Nellore bulls

O objetivo deste estudo foi identificar proteínas ligadoras à heparina no plasma seminal de touros Nelore (Bos taurus indicus). Para tanto, foram selecionados quatro touros entre 30 e 36 meses de idade e peso aproximado de 500-550kg. Após centrifugação, amostras do plasma seminal foram misturadas e as proteínas ligadoras à heparina foram isoladas por meio da cromatografia por afinidade. As frações após a eluição foram agrupadas para caracterização das bandas protéicas (SDSPAGE, 12,5%). Foram identificadas oito bandas protéicas variando entre 15 e 63kDa. Duas proteínas com 22 e 25kDa foram similares às descritas em touros Bos taurus taurus. Outras proteínas identificadas com 39, 53, 58 e 63kDa ainda não foram descritas e possivelmente sejam específicas para Bos taurus indicus.The aim of this study was to identify heparin-binding proteins (HBPs) in seminal plasma of Nellore (Bos taurus indicus) bulls. Bulls (n=4), 30-36 months old, 500-550kg with satisfactory seminal quality were selected. After the centrifugation, samples of the seminal plasma were pooled and the HBPs were isolated by heparin-affinity chromatography. The recovered HBPs fractions were pooled. One-dimensional sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDSPAGE) 12.5% was performed in vertical minigels. Eight bands with molecular weights ranging from 15 to 63kDa were observed. Two proteins were identified (22 and 25kDa), similar to those previously described in Bos taurus taurus bulls. Other bands identified in this study (39, 53, 58 and 63kDa) have not been previously observed and possibly they are specific to Nellore semen

Heparin-binding proteins of seminal plasma in Nellore bulls

IV RESUMO O objetivo deste estudo foi identificar proteínas ligadoras à heparina no plasma seminal de touros Nelore (Bos taurus indicus have not been previously observed and possibly they are specific to Nellore semen

Alpha-glucosidase promotes hemozoin formation in a blood-sucking bug: An evolutionary history

Background: Hematophagous insects digest large amounts of host hemoglobin and release heme inside their guts. In Rhodnius prolixus, hemoglobin-derived heme is detoxified by biomineralization, forming hemozoin (Hz). Recently, the involvement of the R. prolixus perimicrovillar membranes in Hz formation was demonstrated. Methodology/Principal Findings: Hz formation activity of an α-glucosidase was investigated. Hz formation was inhibited by specific α-glucosidase inhibitors. Moreover, Hz formation was sensitive to inhibition by Diethypyrocarbonate, suggesting a critical role of histidine residues in enzyme activity. Additionally, a polyclonal antibody raised against a phytophagous insect α-glucosidase was able to inhibit Hz formation. The α-glucosidase inhibitors have had no effects when used 10 h after the start of reaction, suggesting that α-glucosidase should act in the nucleation step of Hz formation. Hz formation was seen to be dependent on the substrate-binding site of enzyme, in a way that maltose, an enzyme substrate, blocks such activity. dsRNA, constructed using the sequence of α-glucosidase gene, was injected into R. prolixus females' hemocoel. Gene silencing was accomplished by reduction of both α-glucosidase and Hz formation activities. Insects were fed on plasma or hemin-enriched plasma and gene expression and activity of α-glucosidase were higher in the plasma plus hemin-fed insects. The deduced amino acid sequence of α-glucosidase shows a high similarity to the insect α-glucosidases, with critical histidine and aspartic residues conserved among the enzymes. Conclusions/Significance: Herein the Hz formation is shown to be associated to an a-glucosidase, the biochemical marker from Hemipteran perimicrovillar membranes. Usually, these enzymes catalyze the hydrolysis of glycosidic bond. The results strongly suggest that α-glucosidase is responsible for Hz nucleation in the R. prolixus midgut, indicating that the plasticity of this enzyme may play an important role in conferring fitness to hemipteran hematophagy, for instance. © 2009 Mury et al

A lipidomics approach in the characterization of zika-infected mosquito cells: potential targets for breaking the transmission cycle

Recent outbreaks of Zika virus in Oceania and Latin America, accompanied by unexpected clinical complications, made this infection a global public health concern. This virus has tropism to neural tissue, leading to microcephaly in newborns in a significant proportion of infected mothers. The clinical relevance of this infection, the difficulty to perform accurate diagnosis and the small amount of data in literature indicate the necessity of studies on Zika infection in order to characterize new biomarkers of this infection and to establish new targets for viral control in vertebrates and invertebrate vectors. Thus, this study aims at establishing a lipidomics profile of infected mosquito cells compared to a control group to define potential targets for viral control in mosquitoes. Thirteen lipids were elected as specific markers for Zika virus infection (Brazilian strain), which were identified as putatively linked to the intracellular mechanism of viral replication and/or cell recognition. Our findings bring biochemical information that may translate into useful targets for breaking the transmission cycle1110CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQCOORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPSem informaçãoSem informação2011/50400-0; 2015/06809-1; 2014/00084-2; 2014/00302-0; 2013/11343-