RESUMO
The laboratory verified cases of Crimean-Congo hemorrhagic fever (CCHF) in the piedmont steppes of the North Caucasus (Malgobeksky District, Republic of Ingushetia) are first described. The source of the first infection was Ixodidae ticks; three subsequent sources were contacts with the bloody discharges from patients. CCHF virus genome was detected in the blood of the cattle from an epidemic focus and in the pools of the Ixodes ticks Haemaphysalis parva Neum., 1897 and Boophilus annulatus Say, 1821, taken from cattle. The problem of including the piedmont steppes of the North Caucasus into the CCHF nosological area is discussed.
Assuntos
Surtos de Doenças , Reservatórios de Doenças/veterinária , Transmissão de Doença Infecciosa , Vírus da Febre Hemorrágica da Crimeia-Congo/isolamento & purificação , Febre Hemorrágica da Crimeia/epidemiologia , Adulto , Idoso , Animais , Anticorpos Antivirais/sangue , Bovinos , Feminino , Vírus da Febre Hemorrágica da Crimeia-Congo/genética , Vírus da Febre Hemorrágica da Crimeia-Congo/imunologia , Febre Hemorrágica da Crimeia/diagnóstico , Febre Hemorrágica da Crimeia/transmissão , Humanos , Transmissão de Doença Infecciosa do Paciente para o Profissional , Ixodidae/virologia , Pessoa de Meia-Idade , Morbidade , RNA Viral/sangue , Federação Russa/epidemiologiaRESUMO
Trypsin PC from the hepatopancreas of the king crab Paralithodes camtschatica was isolated and purified to apparent homogeneity by ion-exchange chromatography on Aminosilochrom and DEAE-Sephadex and affinity chromatography on arginine-agarose. The yield of the enzyme was 37.7%, and the purification degree was 21. Trypsin PC has a molecular mass of 29 kDa and pI < 2.5. It hydrolysis N-benzoyl-L-arginine p-nitroanilide at the optimum pH of 7.5-8.0 and at the temperature optimum of 55 degrees C (K(m) = 0.05 mM). Trypsin PC retained its activity within the pH range of 5.8-9.0 in the presence of Ca2+. The enzyme was inhibited by the specific inhibitors of serine proteases diisopropyl fluorophoshate and phenylmethylsulfonyl fluoride, by the trypsin inhibitor N-tosyl-L-lysylchloromethylketone, and by the trypsin inhibitors from soybean and potato. Trypsin PC was found to hydrolyze amide bonds formed by carboxylic groups of lysine and arginine in peptide substrates. The N-terminal sequence of this enzyme is IVGGTEVTPG.
Assuntos
Braquiúros/enzimologia , Tripsina/isolamento & purificação , Sequência de Aminoácidos , Animais , Bovinos , Cromatografia de Afinidade , Cromatografia por Troca Iônica , Colágeno/metabolismo , Eletroforese em Gel de Poliacrilamida , Estabilidade Enzimática , Fibrinogênio/metabolismo , Concentração de Íons de Hidrogênio , Ponto Isoelétrico , Fígado/enzimologia , Peso Molecular , Pâncreas/enzimologia , Homologia de Sequência de Aminoácidos , Especificidade por Substrato , Tripsina/química , Tripsina/metabolismo , Inibidores da Tripsina/farmacologiaRESUMO
Various methods for obtaining oxytocin from its recombinant precursor oxytocinoyllysine were studied. For splitting off the C-terminal lysine residue in oxytocinoyllysine, the carboxypeptidase B and an analogous carboxypeptidase from king crab hepatopancreas were used. Ammonolysis of oxytocinic acid methyl ester proved to be the most efficient method for the last stage of the oxytocin preparation. Reversed-phase HPLC was used for the product analysis at each stage of the recombinant oxytocinoyllysine conversion into oxytocin.
