[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.

[Migration of 51Cr-labeled neutrophils to rat lungs after administration of glucoocorticoid kenalog].

Cells from the rat bronchoalveolar lavage fluid comprising over 70% of neutrophils were labeled with 51Cr and administered intraperitoneally or intravenously to control animals and to rats which were injected subcutaneously with 2 mg of ketalog 5 days before the onset of experiment. Blood samples, lung, hepatic, splenic, renal, small intestinal, and muscular biposies were taken 1 hour after cell infusion. The intraperitoneal administration of 51Cr-labeled neutrophils did not significantly increase the radioactivity count rate in any organ examined. The rise was most notable in the lungs after v/v infusion of labeled neutrophils. Rats given kenalog showed a low level of radioactivity in the lung tissue compared with controls probably because the glucocorticoid caused a specific change in the mechanism of uptake of labeled neutrophils.

[Studies of properties of pandemic influenza virus A/H1N1 circulated in Russian Federation in 2009 - 2010].

AIM: Studies of cultural, virologic, antigenic properties of 89 samples of pandemic influenza A(H1N1) 2009 virus isolated in Russian Federation from May 2009 to March 2010. MATERIALS AND METHODS: Properties of isolated samples were compared with those of the reference strain A/ California/04/2009 (H1N1). RESULTS: Studies of biological properties and analysis of genome nucleotide sequences of the isolated samples showed that those strains are closely related to the reference strain. CONCLUSION: Monitoring of genetic, virologic and antigenic properties of pandemic influenza A(H1N1) 2009 virus isolates carried out from May 2009 to March 2010 did not reveal significant changes in the abovementioned properties of the virus or emergence of mutations that can lead to such changes.

[Influence of a glucocorticoid kenalog on functional activity of macrophages and neutrophils both in lungs and in peritoneal cavity].

The work purpose was comparative studying the number change and functional activity of macrophages and neutrophils both in lungs and in peritoneal cavity under the influence of a synthetic glucocorticoid kenalog (Kn). Kn was entered to rats Wistar unitary subcutaneously in a dose of 2 mg/kg (experience), the control rats obtained similarly 0.9% solution NaCl. The cell number of rat bronchoalveolar lavage liquid (BALL) and peritoneal lavage liquid (PLL) was defined in different terms after that. In cell monolayers of BALL we estimated ability of alveolar macrophages (AMph) and neutrophils to engulf of Staphylococcus and to generate of superoxide-anionradicals. In cell monolayers of PLL we estimated the functional activity of peritoneal macrophages (PMph) and neutrophils by means of the same tests. Through 6 (see symbol) after introduction Kn in BALL there was the reduction, and through 5 days--the increase of total cell number with the maximum of neutrophil accumulation and their considerable prevalence over AMph on number, phagocyte and superoxide-producing activity. At the same time the cell number in PLL was decreased and reached a minimum for 5 days after Kn-introduction with considerable decrease of phagocyte and superoxide-producing ability both PMph and neutrophils. The differing action of glucocorticoids on macrophages and neutrophils of different organism compartments and recruitment of considerable quantity of neutrophils, possessing in destructive potential, into lungs can serve as an explanation of many cases when high-dose glucocorticoid therapy is not effective and may be even harmful at attempts of preventing acute respiratory distress-syndrome.

[Study of efficacy of anaferon pediatric in mice infected by pandemic influenza virus A(H1N1/09)v].

AIM: To study efficacy of anaferon pediatric in mice infected by pandemic influenza virus A(H1N1/09)v. MATERIALS AND METHODS: Influenza virus strain A/California/07/2009 (H1N1)v was used. Three groups of BALB/c mice intranasally inoculated with influenza virus were studied. First group received solution of Anaferon pediatric during 5 days before and 8 days after inoculation, 2nd group received Tamiflu during 5 days after inoculation. Distilled water was administered orally to mice from control group. RESULTS: It was shown that Anaferon pediatric used as preventive and treatment agent in mice intranasally inoculated with 100% infectious dose of influenza virus strain A/ California/07/2009 (H1N1)v had antiviral effect, which expressed in 10-fold decreased reproduction of influenza virus in lungs of infected mice compared to control group measured 4, 6, and 8 days after inoculation. CONCLUSION: Use of anaferon pediatric before and after inoculation with influenza virus A(H1N1/09)v was not less effective than use of Tamiflu after inoculation.

[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 antiviral activity of extracts obtained from basidial fungi against influenza viruses of different subtypes in experiments in vitro and in vivo].

AIM: To study antiviral activity of extracts obtained from basidial fungi against influenza viruses of different subtypes. MATERIALS AND METHODS: Antiviral activity of extracts obtained from basidial fungi against influenza virus A/chicken/Kurgan/05/2005 (H5N1) was determined in in vitro experiments. Changes in infectiousness of pandemic influenza virus A/Moscow/226/2009 (HIN1)v caused by extracts of basidial fungi was studied in experiments in vitro and in vivo. RESULTS: Seventy water extracts of basidial fungi were studied, of which 10 were able to inhibit infectiousness of influenza virus strain A/ chicken/Kurgan/05/2005 (H5N1) in MDCK cell culture. Also, several studied extracts decreased infectiousness of pandemic influenza virus strain A/ Moscow/226/2009 (H1N1)v in MDCK cells and inhibit its reproduction in lungs of infected mice. CONCLUSION: High antiviral activity of extracts obtained from basidial fungi against influenza viruses opens perspectives for development of drugs with preventive and treatment effects.

[Entification of the Rubella virus genotype 1H in Western Siberia].

Molecular epidemiological study of novel strain of Rubella virus isolated during the outbreak in Western Siberia in 2004 was described. Detailed phylogenetic analysis performed based upon entire SP-region, which encodes all three Rubella structural proteins (C, E2, and E1), was implemented. This analysis provides characterization of this strain and classifies it as 1H genotype, thereby correcting previous classification of this strain based upon shorter nucleotide sequence, only encoding E1 protein. Therefore, this study identified the genotype of the Rubella virus not previously detected in Western Siberia (and even entire Russian Federation), which highlights the importance of more extensive characterization of genetic variability of the Rubella virus, especially with regard to potential influence of vaccination on the Rubella virus mutagenesis.

[Characterization of the H5N1 influenza virus isolated during an outbreak among wild birds in Russia (Tuva Republic) in 2010].

The study of basic biological properties of H5N1 subtype strain isolated during an outbreak among wild birds in Russia in 2010 was presented. The study was carried out using conventional methods according to the WHO recommendations. H5N1 influenza virus isolated in Siberia belonged to clade 2.3.2 of the hemagglutinin gene; the phylogenetic analysis was performed. The antigenic characteristics and the basic genetic markers of biological properties were studied. It was shown that all strains were highly pathogenic for chickens and white mice. Thus, it was shown that in Russia in the 2010 H5N1 virus phylogenetically closely related to Asian variants caused epizootic among wild birds. The potential danger of this variant of the virus for humans was confirmed by different methods. We discussed the possibility of formation of H5N1 influenza natural focus.

Antiviral activity of Anaferon (pediatric formulation) in mice infected with pandemic influenza virus A(H1N1/09).

Anaferon (pediatric formulation) administered in the therapeutic-and-prophylactic regimen to mice receiving intranasally 100% infecting dose of A/California/07/2009(H1N1)v influenza virus exhibited an antiviral effect and 10-fold reduced the production of influenza virus in the lungs of infected mice on days 4, 6, and 8 after infection compared to the control (distilled water). The efficiency of Anaferon (pediatric formulation) administered before and after infection with A/California/07/2009(H1N1)v influenza virus was not inferior to the use of Tamiflu after infection.

[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).

[Comparative analysis of the susceptibility and productivity of respiratory tract target cells of mice and rats exposed to inflienza virus in vitro].

The levels of susceptibility to influenza virus A/Aichi/2/68 H3N2 and the virus yield were determined using primary cells of the trachea and lungs of CD-1 mice and Wistar rats, and for 3 sets of cells obtained from primary lung cells of the both species by centrifugation in the gradient of density and by sedimentation on a surface. The values of ID50 virus dose for 10(6) cells and virus yield per 1 infected cell determined for primary mice cells were 4.0+/-0.47 and 3.2+/-0.27 IgEID50 (lung cells), 3.8+/-0.17 and 3.3+/-0.20 IgEID50 (tracheal cells), and those determined for primary rat cells were 4.0+/-0.35 and 2.1+/-0.24 IgEID50 (lung cells), 3.7+/-0.27 and 2.2+/-0.46 IgEID50 (tracheal cells). The values of ID50 and yield measured for mixtures of cells obtained from primary lung cells by centrifugation in gradient of density and by sedimentation on a surface differed insignificantly (p = 0.05) from the values of the corresponding parameters measured for lung and tracheal cells for both rats and mice. The analysis of data on the variation of the concentrations of different cell types in the experimental cell mixtures shows that type 1 and 2 alveolocytes possess significantly lower (p = 0.05) susceptibility and productivity vs. ciliated cells of the both species. The investigation was conducted within the frame of the ISTC/DARPA#450p project.

[Analysis of genomes of two rubella virus strains from the 2004-2005 outbreaks in West Siberia].

Two outbreaks of rubella infections notified in the Tomsk and Kemerovo Regions were investigated. Two rubella virus strains from one patient in each outbreak were isolated and genetically characterized. Reverse transcription polymerase chain reaction was used to reveal partial E1 gene sequence at a length of 915 nucleotides. Analysis indicated that the rubella virus strains circulating in the West-Siberian region belonged to international genetic 1g group, which had been first detected in Russia.

[Clinical and immunological characteristics of rubella in western Siberia].

Evaluations of immune system of 155 patients with rubella and 90 contacts with patients were examined. Detection of viral genetic material in blood, urine, and nasopharyngeal swabs has been performed using RT-PCR method. Clinical diagnosis has been confirmed by RT-PCR in 114 (73.5%) patients. Changes of laboratory tests for rubella without clinical signs of the infection were observed in 20% of contacts. Complex ELISA- and PCR-assisted examination of patients can help to determine the stage of disease and characteristics of immune response. For differential diagnostic of rubella and other infectious diseases with exanthema it is rational to perform complex examination of patients using immunologic and molecular biologic methods.

[Genotyping of rubella virus circulating in Western Siberia of Russia during 2004-2006 epidemic period].

Twenty one strains of rubella virus were isolated in the Western Siberia during 2004-2006 epidemic period. Genotyping of isolated strains was performed by partial sequencing of glycoprotein E1 gene. Phylogenetic analysis showed that 20 out of 21 isolated in the Western Siberia strains of rubella virus belonged to genotype 1g, and 1 strain (isolated in Altai region in 2006)--to genotype 1E.

Comparative study of pathological changes in the mouse viscera in infection caused by two orthopoxviruses.

The mechanisms of infection development in intraperitoneal inoculation of mice by ectromelia virus strain K-1 and cowpox strain EP-2 were studied. Ultrastructural parameters of virus assembly and maturation are described. Differences in the types of cells replicating the viruses and in the type of visceral injuries were detected. The studies showed a local type of strain EP-2 cowpox infection and dissemination of ectromelia strain K-1.

[The influence of glucocorticoid immunosuppression on the course of experimental influenza pneumonia].

The study demonstrates the effects of kenalog (Kn), a synthetic glucocorticoid hormone, on the course of virus A/Aichi/2/68 influenza in white mice. In doses of 5 and 10 mg/kg, Kn reduced the weight of the adrenal glands, thymus and spleen, which was accompanied by decrease of the resistance to the mentioned virus, judging by LD50 decrease vs. this index in the control infected group. Besides, four days after infecting with 5 LD50 of influenza virus (IV), lung virus and interferon titers were significantly lower in mice pretreated with Kn vs. mice treated with placebo. Lung cell susceptibility to IV in vitro was identical in mice treated with Kn or placebo. In ultrathin lung sections of IV-infected mice, both experimental and control ones, there was virus budding in bronchial epithelium cells and type I and II alveolocytes. Analysis of inflammatory effusion compound in semithin lung sections 6 days after IV infection, found a substantially smaller number of mature alveolar macrophages (AM) and a bigger number of neutrophiles vs. infected controls. The authors reckon that higher mortality of mice pretreated with Kn before infecting, is caused not by enhancement of IV reproduction in target lung cells during influenza development, but by the contribution of other pathogenic factors. One of those may be increase of neutrophilic migration into the lungs; neutrophiles are more able to realize their significant destructive potential under the condition of reduction in the clearing function of AM and IV infection.

[The morphology of the micro-capsulated polyacrylic acid-based formulation of measles vaccine].

The morphology and virus localization were studied in the microcapsulated measles vaccine formulation involving polyacrylic acid (PAA) copolymers as a matrix. Transmission electron microscopy and phosphotungsic staining at a pH value of 2 to 7 showed that the morphology of microparticles was related to the value of pH and to the concentration of a polymer in the matrix. In the neutral medium, the microcapsules had the sizes of 0.5 to 10 microm, which were optimal for transport through the intestinal wall when immunization was orally used. Immunocytochemistry revealed measles virus antigen within the microparticles. The specific activity of the microcapsulated formulation of measles virus was as high as 3.36 and 4.31 lg of TCD50/0.5 ml for the samples containing 1 and 0.1% polymer, respectively. The findings suggest that the microparticles of the vaccine contain live measles virus.