RESUMO
Glycosidase activities were detected as detergent-insoluble after sequential extractions of goat sperm with Triton X-100. Seventy percent of total beta-glucuronidase activity was found in the detergent-insoluble fraction. This portion of beta-glucuronidase was resistant to extractions in the presence of 1 M KCl, chaotropic agents, colchicine, or cytochalasin B, being only partially solubilized by 3 M KCl or DNAse I treatment. The treatment with 0.1% sodium deoxycholate was effective, releasing 73% of the enzyme activity. Treating the deoxycholate extract with DNAse I resulted in a change in the elution profile of beta-glucuronidase as judged by gel filtration chromatography. A polyclonal antibody was developed against pancreatic beta-glucuronidase, and the sperm enzyme was strongly inhibited by the IgG fraction of this antibody. Western blot analysis showed that the same protein correspond to both Triton-soluble and insoluble enzyme. Results demonstrate that beta-glucuronidase is tightly bound to the Triton X-100 resistant fraction, suggesting that the enzyme is associated to sperm cytoskeleton. J. Exp. Zool. 289:146-152, 2001.
Assuntos
Citoesqueleto/enzimologia , Glucuronidase/metabolismo , Espermatozoides/enzimologia , Animais , Cabras , Masculino , Octoxinol , Espermatozoides/citologiaAssuntos
Vacinas Bacterianas/administração & dosagem , Vacinas Bacterianas/efeitos adversos , Vacinas contra Influenza/administração & dosagem , Vacinas contra Influenza/efeitos adversos , Infecções Pneumocócicas/prevenção & controle , Adulto , Idoso , Brasil , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
Affinity chromatography on Concanavalin-A Sepharose, followed by gel filtration and hydrophobic interaction chromatography, permits the isolation of low molecular weight N-glycosidically linked oligomannosidic glycopeptides (MGp) from the autoproteolysis products of human seminal plasma. The monosaccharide composition of MGp showed only mannose, N-acetylglucosamine and a small amount of galactose. Structural studies were carried out by methylation analysis and chromium trioxide oxidation, and results were consistent with the structures accepted for high-mannose N-glycans. MGp was capable of inhibiting the sperm acrosomal exocytosis mediated by sperm-surface receptors. These data suggest that MGp act as a "decapacitation" factor preventing premature sperm exocytosis.
Assuntos
Glicopeptídeos/isolamento & purificação , Sêmen/química , Reação Acrossômica/efeitos dos fármacos , Configuração de Carboidratos , Cromatografia de Afinidade , Cromatografia Gasosa , Cromatografia em Gel , Exocitose , Glicopeptídeos/química , Glicopeptídeos/farmacologia , Humanos , Masculino , Manose/química , Oligossacarídeos/químicaRESUMO
The need to understand the meaning of parenteral nutrition to the client in hospital context has come from my personal questioning on nursing care. Participative observations have been made as well as interviews and diligences with seven clients in hospital, who have been having parental nutrition. These information have been analysed through interactive approach, showing the situation faced by patients who have hospital assistance; how they say parental nutrition within this context; and what meaning has been given to it. This study revealed feelings, behaviours and attitudes related to the biological, emotional and social cultural aspects of the patient. All of them have been linked to parental nutrition. The limits faced by parental nutrition and its obstacles in the nursing assistance were finally understood.