RESUMO
The modern influenza virus subtypes H3N2, H5N1, and H1N1 reduced the metabolism of the endothelial cells within the range from 20% to 60% (compared with control). The degree of the activity of the dehydrogenase reduction depended on the dose of virus and time of virus reproduction. HA and NA also actively reduced the metabolism of the cells ranging from 5% to 60%, depending on the concentration of the proteins and time of their impact on cells. Neuraminidase was more active than hemagglutinin in the MTT test (at concentration 50 microg protein/ml).
Assuntos
Células Endoteliais/efeitos dos fármacos , Endotélio Vascular/efeitos dos fármacos , Glicoproteínas de Hemaglutininação de Vírus da Influenza/farmacologia , Vírus da Influenza A Subtipo H1N1/crescimento & desenvolvimento , Vírus da Influenza A Subtipo H3N2/crescimento & desenvolvimento , Virus da Influenza A Subtipo H5N1/crescimento & desenvolvimento , Neuraminidase/farmacologia , Biomarcadores/metabolismo , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Células Endoteliais/citologia , Células Endoteliais/virologia , Endotélio Vascular/citologia , Endotélio Vascular/virologia , Glicoproteínas de Hemaglutininação de Vírus da Influenza/isolamento & purificação , Humanos , Vírus da Influenza A Subtipo H1N1/patogenicidade , Vírus da Influenza A Subtipo H3N2/patogenicidade , Virus da Influenza A Subtipo H5N1/patogenicidade , Neuraminidase/isolamento & purificação , Oxirredutases/antagonistas & inibidores , Oxirredutases/metabolismo , Sais de Tetrazólio , TiazóisRESUMO
Cold-adapted influenza virus A/HK/1/68/162/35(H3N2) was developed as unified donor of attenuation and high reproductive capacity forvaccine strains. The reassortant of this donor with surface antigens of highly pathogenic strain Alchicken/Astana/6/05 (H5N1) was tested in guinea pigs as a live or inactivated preparation. Immunization with both formulations induced equal levels of serum virus specific antibodies, while the level of mucosal antibodies was significantly higher in animals immunized with live virus. The challenge with the homologous virus demonstrated complete virus clearance only in this group, thereby indicating a direct correlation of the protection level with the level of mucosal antibodies.
Assuntos
Anticorpos Antivirais/imunologia , Vírus da Influenza A Subtipo H2N2/imunologia , Virus da Influenza A Subtipo H5N1/imunologia , Vacinas contra Influenza/imunologia , Neuraminidase/imunologia , Infecções por Orthomyxoviridae/prevenção & controle , Vírus Reordenados/imunologia , Adaptação Biológica , Animais , Anticorpos Antivirais/sangue , Temperatura Baixa , Cobaias , Imunidade nas Mucosas , Imunização , Vírus da Influenza A Subtipo H2N2/genética , Virus da Influenza A Subtipo H5N1/genética , Vacinas contra Influenza/administração & dosagem , Neuraminidase/genética , Testes de Neutralização , Infecções por Orthomyxoviridae/imunologia , Infecções por Orthomyxoviridae/virologia , Vírus Reordenados/genética , Mucosa Respiratória/imunologia , Mucosa Respiratória/virologia , Vacinas AtenuadasRESUMO
AIM: Detection of antibodies against neuraminidase (NA) of A/California/07/2009 (H1N1) influenza virus in blood sera of volunteers after the immunization with live monovalent influenza vaccine (LIV). MATERIALS AND METHODS: Neuraminidase enzyme activity inhibition by antibodies test with reassortant strain A(H7N1) containing NA of pandemic strain was used. Anti-neuraminidase IgG antibodies against whole reassortant virus A(H7N1) and purified NA of A/California/07/2009 (H1N1) strains were determined by enzyme immunoassay (EIA). RESULTS: After two immunizations with LIV of seronegative individuals a 1.5 times mean increase of antibodies against homologous neuraminidase was detected (by hemagglutinin inhibition reaction). The high level of anti-neuraminidase antibodies were detected in individuals that had been naturally infected. A correlation between anti-neuraminidase IgG antibody titers obtained in EIA with whole reassortant virus A(H7N1) and purified protein was demonstrated. CONCLUSION: Modified sialidase activity inhibition method and EIA with reassortant diagnostic strain can be applied to evaluate anti-neuraminidase antibodies.
Assuntos
Anticorpos Antivirais/biossíntese , Vírus da Influenza A Subtipo H1N1/isolamento & purificação , Vírus da Influenza A Subtipo H7N1/isolamento & purificação , Vacinas contra Influenza/administração & dosagem , Influenza Humana/prevenção & controle , Neuraminidase/imunologia , Vírus Reordenados/isolamento & purificação , Vacinação , Adolescente , Animais , Anticorpos Antivirais/análise , Formação de Anticorpos/efeitos dos fármacos , Formação de Anticorpos/imunologia , Antígenos Virais/imunologia , Embrião de Galinha , Ovos/virologia , Testes de Inibição da Hemaglutinação , Humanos , Imunoensaio , Vírus da Influenza A Subtipo H1N1/genética , Vírus da Influenza A Subtipo H1N1/imunologia , Vírus da Influenza A Subtipo H7N1/genética , Vírus da Influenza A Subtipo H7N1/imunologia , Influenza Humana/imunologia , Influenza Humana/virologia , Vírus Reordenados/genética , Vírus Reordenados/imunologia , Vacinas Atenuadas/administração & dosagem , Replicação Viral , Adulto JovemAssuntos
Surfactantes Pulmonares/farmacologia , Síndrome do Desconforto Respiratório/tratamento farmacológico , Líquido Amniótico/química , Animais , Gasometria , Líquido da Lavagem Broncoalveolar/química , Bovinos , Cães , Feminino , Humanos , Pulmão/química , Pulmão/efeitos dos fármacos , Pulmão/patologia , Camundongos , Microscopia Eletrônica , Fosfolipídeos/análise , Gravidez , Surfactantes Pulmonares/química , Surfactantes Pulmonares/toxicidade , Ratos , Síndrome do Desconforto Respiratório/patologiaRESUMO
A novel procedure for purification of copper-thionein from rabbit liver was developed. The molecular mass, copper, and thiol contents were determined. The properties of the protein were compared with those of Cd-thionein. Copper was removed from Cu-thionein and the reconstitution was achieved. The binding of copper leads to an increase in absorbance at 260 nm.
Assuntos
Fígado/análise , Metalotioneína/isolamento & purificação , Animais , Apoproteínas/análise , Apoproteínas/isolamento & purificação , Cromatografia em Gel , Cobre/análise , Metalotioneína/análise , Peso Molecular , Coelhos , Espectrofotometria Atômica , Compostos de Sulfidrila/análiseRESUMO
Superoxide dismutases (SOD) of high purity have been obtained from grey and white matter of bovine brain cerebral hemispheres. The SOD obtained have been shown to have three isoenzymes (a, b, c). A study of the catalytic and macromolecular properties of the SOD obtained from grey and white matter of cerebral hemispheres as well as their optical and electron paramagnetic resonance spectra indicates that both proteins possess similar properties. The possible function of SOD in grey and white matter of brain is discussed.
Assuntos
Encéfalo/enzimologia , Superóxido Dismutase , Animais , Catálise , Bovinos , Espectroscopia de Ressonância de Spin Eletrônica , Isoenzimas/isolamento & purificação , Cinética , Peso Molecular , Análise Espectral , Superóxido Dismutase/antagonistas & inibidores , Superóxido Dismutase/isolamento & purificaçãoRESUMO
Cytochrome c from grey matter of brain has been obtained as a homogeneous preparation by electrophoresis on polyacrylamide gel and following electrofocusing in ampholine solutions. Its molecular weight, content of iron, redox potential and isoelectric point have been established. The absolute spectra of its oxidized and reduced forms are presented. Cytochrome c of brain cortex is similar in its properties to that obtained from other animal tissues as the heart and adrenal cortex.