[Molecular-biological and antigenic features of H5N1 subtype highly pathogenic influenza virus strains isolated in southern Siberia in 2005-2009].

AIM: Study of molecular-biological and antigenic features of H5N1 subtype virus strains isolated in southern Siberia in 2005-2009. MATERIALS AND METHODS: Study was performed by using standard procedures according to WHO recommendations. RESULTS: Hemagglutinin gene of H5N1 subtype virus strain isolated in Siberia belongs to 2 genetical lineages: 2.2 and 2.3.2. Strains of these 2 lineages have antigenic differences. All of the strains are highly pathogenic for chicken and white mice. CONCLUSION: H5N1 subtype highly pathogenic influenza virus variants of 2 different antigenic lineages that have antigenic differences circulated in Siberia in 2005-2009. A possible role of antigenic drift in evolution of H5N1 subtype influenza virus is discussed.

[Genetical features of influenza virus A (H1N1) strain that caused the 2009 pandemic].

Genetical features of the A(H1N1) influenza virus strain that caused the 2009 pandemic are analyzed in the review. Mutations typical for this strain, unique and similar to influenza viruses of swine, avian and seasonal types, and phenotypic (pathologic) features associated with them, that are experimentally confirmed, are described. A possibility of reassortation of avian and swine influenza viruses and possible epidemiologic consequences are discussed.

[Pandemic influenza virus A (H1N1) in Amur region in autumn 2009].

AIM: Isolation and study of molecular genetic characteristics of pandemic influenza virus A (H1N1) circulated in Amur region in autumn 2009 as well as testing of serum samples taken from citizens of this region during November- December 2009 in order to measure levels of antibodies to socially significant serotypes of influenza A virus. MATERIALS AND METHODS: Strain of pandemic influenza virus A/Blagoveschensk/01/2009 (H1N1) was isolated on MDCK cell culture and nucleotide sequences of all eight segments of viral genome were determined. Five hundred seventy-six serum samples taken in Amur region in autumn 2009 were tested by hemagglutination inhibition assay. RESULTS: Nucleotide sequence of A/Blagovechensk/01/2009 (H1N1) strain was 99.7% identical to reference influenza virus strain A/California/04/2009. Diagnostically significant titers of antibodies to pandemic influenza virus were observed in 46.3% of persons younger 30 years old and in 20.1% older persons. Antibodies to seasonal influenza virus H1N1 and H3N2 were detected in 39.5 and 29.8% of persons respectively. CONCLUSION: Final seroepidemiological picture of distribution of pandemic virus in Amur region matches with the one for seasonal influenza virus A (H1N1): > 60% of seropositive persons were registered in age group < 18 years old, and this proportion increases with increasing age.

[Study of the antiviral activity of Russian anti-influenza agents in cell culture and animal models].

The study of the antiviral activity of Russian anti-influenza agents in the cultured MDCK cells demonstrated that arbidol and ribavirin inhibited the reproduction of various influenza A virus strains, including rimantadine- and ozeltamivir-resistant variants, as well as influenza B viruses (IC50 2-8.5 microg/ml). Rimantadine at concentrations of 1-5 microg/ml completely inhibited the reproduction of reference and ozeltamivir-resistant influenza A virus strains, and it had no effect on the reproduction of influenza B viruses and rimantadine-resistant influenza A viruses. Arbidol and ribavirin also inhibited the reproduction of pandemic influenza A/California/04/2009(H1N1), A/California/07/2009(H1N1), and A/Moscow/01/2009(H1N1)swl viruses in the cultured MDCK cells (IC50 = 1.5-4.0 microg/ml) while rimantadine had no effect on their reproduction. The cultured cells showed no significant antiviral activity of ingavirin at nontoxic concentrations (up to 200 microg/ml) against all study strains of influenza A and B viruses, including pandemic A(H1N1) influenza virus strains. The activity of rimantadine, arbidol, and ingavirin was found on a model of Influenza pneumonia in mice infected with their adopted influenza A/Aichi/2/69(H3N2) virus. The preventive efficacy of the three test agents was similar and most pronounced when they were used 96 hours before infection, by preventing 40-50% death in the animals and their body weight loss and by increasing their survival by 1.3-1.5 times. Arbidol and rimantadine were more effective when used for treatment and prophylaxis in doses of 30 and 10 mg/kg/day, respectively, by protecting the infected animals from 60-80% death, increasing their survival by 1.7-2 times, and preventing their body weight loss as compared with the control. The same experiments with ingavirin showed that this agent was less effective than arbidol and rimantadine. Thus, arbidol and rimantadine have a pronounced antiviral infection in both cell culture and a model of influenza pneumonia. The found efficacy of ingavirin on an integral model of murine influenza pneumonia without its activity in the cultured cells is likely to be due to other pharmacological properties of the drug rather than its direct virus-specific action.

[Monitoring for influenza in population of Western Siberia in 2007-2009].

AIM: To analyze influenza viruses isolated in the 2008-2009 autumn-winter season, and to test sera collected in the south of Western Siberia during the beginning and the end of the epidemic seasons from 2007 until the A/H1N1 pandemic. MATERIALS AND METHODS: Total 149 clinical samples were analyzed and 2190 blood sera were tested. During the 2008-2009 season 17 influenza viruses were isolated. 9 of these were A/H1N1, 5-were A/H3N2, and 3 were influenza B viruses. The nucleotide sequences and amino acid composition of influenza A virus hemagglutinin (HA) were compared with reference strains. RESULTS: Among A/H1N1 viruses circulating in Novosibirsk region three viruses contained four amino acid replacements in antigen sites Ca, Cb and Sb. In A/ H3N2 viruses from Novosibirsk, 2 amino acid substitutions were detected in antigen sites B and E. CONCLUSION: Based on genotyping influenzae epidemic on February to April of 2009 in the south of western Siberia was associated with influenza viruses A/H1N1, A/H3N2, and B. All A/H3N2 influenza virus isolates were variants of reference A/Brisbane/10/2007(H3N2) and A/ H1N1 influenza viruses isolates were similar to reference A/Brisbane/59/2007(H1N1).

[Investigation of susceptibility of influenza viruses A (H1N1), the cause of infection in humans in April-May 2009, to antivirals in MDCK cell culture].

Biological properties of influenza viruses A (H1N1), that were the cause of the infection in humans in April - May 2009, and the action of the Russian antivirals on their reproduction were studied in vitro. The nucleotide sequence in the viruses was determined and followed by detection of the mutations responsible for resistance to the antiinfluenza drugs. The experiments showned that arbidol and ribavirin had a selective inhibitory action on reproduction of the viruses in the MDCK cell culture while rimantadine had no affect on their reproduction. The data were confirmed by the results of the genome analysis in influenza viruses A/California/04/2009(H1N1), A/California/07/2009(H1N1) and A/Moscow/01/2009(H1N1)swl, that revealed no replacements defining the resistance to arbidol while the viruses contained a mutation in position 31 of M2 protein, responsible for the resistance to adamantans.

[Effects of plasminogen, streptokinase and their equimolar complexes with pyruvate kinase on the human neuroblastoma IMR-32 cells].

The system of extracellular proteolysing, consists of plasminogen (PGn), its active protease (plasmin), PGn activation and PGn activators inhibitors, influences the nervous tissue functions, their growth, differentiation and proliferation in both, normal and pathological conditions. The purpose of the investigation was to study the effects of exogenous PGn, its activator streptokinase (SK), PK and their equimolar complex on the morpho-functional state neuroblastoma IMR-32 cells. PGn, SK, PK and their complexes stimulated cells proliferation during 1-3 days of incubation, shown by cell quantity increase. We also observed DNA, RNA and protein increase. The low lactate dehydrogenase efflux was evidence of that an addition of the proteins under investigation in the culture medium prevented the development of degenerative alterations connected with serum deprivation. The levels of extracellular PGn-activator activity, as measured by the biochemical fibrinolytic assay, increased over SK. This SK effect vanished on the 3rd day when SK formed complexes with PK. New original facts obtained testify the probability of initiation of neoplastic transformation and tumor growth potentiation.

[Effects of plasminogen and streptokinase on the vital functions of nervous tissue cells in culture].

In the protein-deficient media plasminogen stimulated the vital functions of cells and in concentrations 10(-7)-10(-10) M it protected cells of sympathetic ganglia, neocortex and continues cell lines under damaging actions of H2O2 (0.0001 M), NH4CI (0.01 M) and cooling. Streptokinase essentially influenced the mode of damaging effect of ATP(0.001 M). Even a short-term exposition (20 min) of PC12 cells with both proteins (each in the concentration 10(-9) M) led to sharp alterations in intracellular ATP- or Ca(2+)-activated proteolysis. In some cases plasminogen and streptokinase provided acceleration of cultured tissue maturation, improvement of cell adhesion, high survival rate, the increase in quantity and length of processes and their arborisation. Electronic microscopy established the character of structural rearrangements of nervous tissue cells (neurons, astrocytes, oligodendrocytes), reflecting the protective action of plasminogen and streptokinase. In the presence of plasminogen and especially streptokinase, the total number of cultured glioma C6 and neuroblastoma IMR-32 cells, the intracellular contents of protein, RNA and DNA increased several-fold. Addition of plasminogen promoted formation of processes by neuroblastoma cells, this suggests initiation of differentiation of cellular elements. In cultures of sensitive and sympathetic ganglia streptokinase increased proliferation of Schwann cells. These proteins did not cause transformation of PC12 enterochromaffine cells to neurons, though plasminogen facilitated it. Plasminogen addition to cell cultures did not increase fibrinolytic activity of the culture medium in the culture medium, and streptokinase did not lose its plasminogen-activating capacity.