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The aim of this investigation was to study the effect of ingavirin on the structure and properties of influenza virions forming in its presence. The infectious activity of the virus and the morphology of the virions were analyzed by titration in cell culture and electron microscopy, respectively. The use of ingavirin was shown to reduce the proportion of morphologically intact virions and to increase that of filamentous and giant particles. No defects of surface glycoproteins were observed. The effect of the drug did not depend on the chosen model of virus replication and it was similarly shown in both cultured human cells and laboratory animals. In MDCK and A549 cells and in the mouse lungs, viral infectious activity was decreased by 1-2 orders of magnitude in relation to a model. The findings suggest that Ingavirin is able to impair the processes of viral morphogenesis, which in turn leads to a reduction in the infectivity of progeny virions.

Currently, a range of biologically active molecules have been attached to plant and bacterial viras nanoscaffolds, yielding stable nanoparticles that display multiple copies of the desired molecule. In this paper we propose a new method of non-covalent attachment of peptides to the surface of virios. We have demonstrated that this method is efficient in a model system that includes tobacco mosaic virus particles, synthetic polycation (quaternized poly(4-vinylpyridine) carrying ethyl ethyl pendant radicals) and polypeptide of interest. This principle of step-by-step binding to the surface of virions was used for electrostatic association with hydrophilic fragment of influenza virus haemagglutinin.

Aim: Characterization of features of influenza pandemic development in Russia in relation to global process., Materials and Methods: Pandemic monitoring was performed by using results of integrative analysis of laboratory diagnostic and population morbidity data from 49 supporting bases of Federal center of influenza from various cities in Russian Federation. Isolation of influenza virus was carried out in MDCK cells and chicken embryos under BSL-3 conditions. Reference virus A/California/07/09 obtained from CDC (Atlanta, USA) and antisera against this strain contained in WHO kit were used for antigenic analysis; rat antisera, new monoclonal antibodies against pandemic influenza virus developed by Research institute of influenza were also used., Results: Based on PCR monitoring during epidemic peak, rate of pandemic influenza identification reached 45-49% of examined patients. About 53% of lethal cases of respiratory infections were caused by pandemic influenza virus, while predominately young people died from pneumonia and acute respiratory distress syndrome. Russian isolates generally were antigenically and genetically similar to the parent pandemic strain--influenza virusA/California/07/09, but contained S203T substitution in hemagglutinin. A number of strains contained D222G mutation that is responsible for the expansion of substrate specificity, as well as strain specific substitutions in hemagglutinin and neuraminidase molecules. The investigated isolates were resistant to remantadin, but sensitive to oseltamivir., Conclusion: Due to the formation of population immunity after the end of the first pandemic wave new drift variants of the virus capable of overcoming this formed immunity should be expected that apparently will require the correction of vaccine composition for the 2011 - 2012 season.

Research Institute of Influenza, Ministry of Health and Social Development of Russia, Saint Petersburg The characteristics of the isolation of pandemic influenza A(H1N1)v viruses were studied on chick embryos (CE) and MDCK cell culture. The materials (nasal swabs and autopsies) were collected in different regions Russia in the period from 20 July to 30 December 2009. The paper gives the data of the antigenic analysis of isolates, their capacity to multiply in different species-specific and tissue cell cultures. The viruses isolated on CE were shown to have higher hemagglutination titers and to be more stable. Isolation from the autopsies was effective only on CE. All the test cell lines other than MDCK were insensitive to the isolated pandemic influenza strains. The antigenic analysis showed no significant antigenic drift of the viruses isolated during the first wave of the pandemic in the Russian Federation.

The paper describes dynamics, distribution and morbidity rate during the 2009 A(H1N1)v influenza epidemic in Russia. The epidemic appears to have been especially severe in the cities of the Far-East and Siberian Federal Districts where the average morbidity rate ranged from 6.4% to 19.2% (mean 10.3%) and the epidemic duration from 7.8 to 8 weeks. In less affected Southern and Central Federal Districts A(H1N1)v influenza occurred in 5.7% of the population. Schoolchildren aged 7-4 years showed the highest morbidity rate of 28.8%. The age group of 18-53 years accounted for 79.4% of the total lethality. Viral isolates were genetically stable and exhibited 98.9% hemagglutnin (HA) homology with reference viruses. None of the strains had an amino acid substitution at position 275 of neuraminidase (NA) responsible for resistance to oseltamivir. Towards the end of the epidemic, the viral population displayed a significant rise in the number of strains containing mutations in 4 genes (4 HA, 2 NA, 2 PB2 and 1 PA mutations respectively). 26.7% of the viral isolates obtained in the end of the epidemic had D222G substitution responsible for tropism of viruses to lung tissues. Epidemiologically, the 2009 A(H1NI)v influenza epidemic is described as moderate based on the absence of pathogenicity determinants typical of both A(H1N1) influenza virus of 1918 and A(H5N1) virus. The paper compares the 2009 epidemic with those caused by A/Honkong/68 and A/USSR/ 90/77 viruses. The necessity of classification for the discrimination between A(H1N1) subtype viruses is emphasized.

The analysis of 1558 clinical samples revealed influenza virus A(H1N1v) RNA in 339 patients with influenza and 163 fatal cases,which was made in May to December 2009. Data on the antigenic properties of more than 250 of pandemic virus strains isolated at the Research Institute of Influenza and the molecular genetic characteristics of 31 strains are presented. All the test isolates were found to have the S203 substitution in hemagglutinin, which was characteristic of one of 5 minor genome A(H1N1v) virus variants found in the United States and Mexico in 2009. All the test strains contain the S31N substitution in the M2 protein, which determines viral resistance to adamantine, and have no H275Y substitution in neuraminidase, which determines oseltamivir resistance. The substitution of amino acid residue of Asp to Gly at position 222 of HA was found in 8 (73%) of 11 isolates from postmortem lung and trachea samples and in 2 (10%) of 20 isolates from nasopharyngeal swabs. The determination of the pathogenic role of this substitution calls for further investigations.

Biological effects of water-soluble inclusion complexes of fullerene C60 with poly(vinyl pyrrolidone) (C60/PVP) and gamma-cyclodextrin (C60/g-CD) as well as solid C60 (C60-coated surface) on cell viability have been studied in vitro. It is established that both inclusion complexes (in a broad range of concentrations) and solid fullerene coatings are nontoxic in the dark for the cell of all lines tested. In contrast, under intense UV illumination, the C60/PVP complex reliably protected test cells from the UV radiation damage, whereas the C60/g-CD and fullerene-coated surface exhibited pronounced phototoxicity. Moreover, solid fullerene caused a photodynamic effect under irradiation with both UV and visible light. The radiation damage could be blocked by some antioxidants (e.g., hypoxen) and singlet-oxygen scavenger (sodium azide). This is evidence for the participation of 1O2 in phototoxicity manifestations. The results indicate that the biological properties of fullerene C60 in vitro depend on its aggregate state, form of solubilization, and, probably, the nature of solubilizing medium.

The effect of meglumine salt of acridonoacetic acid (cycloferon) on the in vivo morphogenesis of influenza infection caused by viruses of different origin (avian, swine and human) and variable susceptibility to antivirals (rimantadine and oseltamivir) has been studied. The administration of cycloferon results in stimulation of the immune response, restriction of the foci of post-influenza pneumonia, and normalization of the structure of respiratory zones independently of the susceptibility or resistance of infectious virus to the drugs. Among virions formed in the lungs of cycloferon-treated mice, prevalence of irregular-shaped virions with defects of surface glycoproteins was observed. The data obtained suggest that cycloferon is a drug with the complex mechanism of activity.

The influence of antivirals, such as rimantadine, ribavirine and triazavirine on influenza virus replication in human cell cultures was evaluated. All the antivirals inhibited viral nucleoprotein NP synthesis. The strongest effect was shown for ribavirine in lung carcinoma A-549 cells and endothelial ECV-304 cells. Hoechst-33258 staining revealed induction of apoptosis in all the cell lines. Rimantadine and ribavirine inhibited virus-induced apoptosis while ribavirine enhanced it. The effect was registered in monolayer cell cultures as well as in suspension cell cultures. The influence of the antiviral drugs on the virus-induced cell proliferation in the suspension cell cultures is also described.

Our study included 112 patients with squamous cell cervical carcinoma la-lib stages (FIGO). All of them were restaged on the basis of histological evidence after surgical treatment. Ninety-eight were staged at final analysis. Human papilloma virus (HPV) was detected in endocervical smears and paraffin blocks of lymph nodes by PCR and real-time PCR. Oncogenic HPV in primary tumor was detected in 86 (87.8%); two or more genotypes--65 (75.58%): still more--21 (24.4%). In the latter group, the frequency of regional lymph node metastases was significantly higher. HPV DNA was identified in iliac lymph nodes in 29 (26.6%); 27 of them had metastases to those nodes. HPV in DNA which was significantly more frequent in involved lymphocytes may be used as marker. Since our method for early detection of metastases to regional lymph nodes is highly specific, it may be recommended as a diagnostic procedure.

The conventional vaccines currently being used to deal with influenza are based on a virus obtained in chicken embryos or its components. The high variability of the major immunogenic surface proteins - hemagglutinin and neuraminidase-require the development of strain-specific vaccines that match the antigenic specificity of a newly emerging virus. Recombinant vaccines based on single viral proteins that could be easily produced in standard expression systems are attractive alternatives to traditional influenza vaccines. We constructed recombinant nanosized virus-like particles based on a nuclear antigen of the hepatitis B virus. These particles expose on the surface the extracellular domain of the M2 protein of the highly pathogenic A(H1N1) 2009 influenza virus. The methods of production of these virus-like particles in Escherichia coli and their purification were developed. Experiments on animals show that M2sHBc particles are highly immunogenic in mice and provide complete protection against the lethal influenza challenge.

Aim: To study tolerability and safety of pandemic vaccines against influenza A/H1N1 "INFLUVIR" and "PANDEFLU" on limited group of volunteers during phase I clinical trial., Materials and Methods: Thirty healthy volunteers were participated in phase I clinical trial. Clinical and laboratory tests of volunteers randomized to 2 groups (20 persons received vaccine and 10 persons - placebo) were performed. "INFLUVIR" vaccine was administered by intranasal route. Volunteers were hospitalized and followed up for development of local and systemic adverse events during 7 days after vaccination. "PANDEFLU" was administered intramuscularly; vaccinees were followed for 7 days in outpatient settings. RESULTS. Phase I clinical trial showed good tolerability and low reactogenicity of "INFLUVIR" and "PANDEFLU" vaccines. There were no moderate and severe local and systemic adverse events registered., Conclusion: On the basis of performed phase I clinical trial, phase II trial was recommended to perform in order to assess the reactogenicity, safety and immunogenicity of studied vaccines.

Chemotherapy and chemoprophylaxis of influenza is one of the most important directions of health protection activity. Due to the high rate of drug-resistant strains of influenza virus, there is a need for the search and further development of new potent antivirals against influenza with a broad spectrum of activity. In the present study, a set of di-, tri- and tetrazole derivatives of adamantane was efficiently prepared and their anti-influenza activities evaluated against rimantadine-resistant strain A/Puerto Rico/8/34. In general, derivatives of tetrazole possessed the highest virus-inhibiting activity. We demonstrated that several compounds of this set exhibited much higher activity than the currently used antiviral rimantadine, a compound of related structure. Moreover, we showed that these azolo-adamantanes were significantly less toxic. This study demonstrates that influenza viruses can be inhibited by adamantyl-azoles and thus have potential for developing antiviral agents with an alternate mechanism of action., (Copyright 2009 Elsevier Ltd. All rights reserved.)

The protective activity of Ingavirin against experimental infection caused by influenza B virus was studied on albino mice vs. Arbidol. Oral use of Ingavirin was shown to decrease the infectious titers of the virus in the animal lung tissue, to normalize the body weight dynamics, to lower the mortality and to increase the average lifespan vs. the placebo-treated animals. The activity of Ingavirin was higher than that of the reference drug. The results allowed to consider Ingavirin as a prospective agent for the treatment of influenza infection in humans.

This article analyzes the current situation in the field of construction and production of pandemic influenza vaccines. The main task of protecting the population against influenza pandemics requires state-of-the-art approaches to the construction of influenza vaccines to be based on reassortment and genetic engineering techniques, including the analysis of primary structures of influenza viral genes, synthesis and cloning of the main viral genes, reverse genetics techniques, and banks of plasmids bearing basic viral genes. Reassortant technologies are now giving way to new approaches for objective reasons. The state-of-the-art technologies provide safety not only at the laboratories where vaccine viruses are constructed but also make the production process wholly safe. We are using the following approaches to the development of industrial production: use of nanoparticles and nanoemulsions as functional adjuvants, construction of totally-safe strains for live attenuated influenza vaccines with deletions of molecular determinants of pathogenicity, application of protein and chemical chaperones to provide self-assembly of haemagglutinin molecules of the H1N1v-2009 virus, and impregnation of whole-virion preparations with nanoparticles to enhance antigenicity., (© Pleiades Publishing, Ltd. 2010.)

Despite obvious success in the vaccine development and chemotherapy of influenza, it remains a poorly controlled infection leading to emergence of new pandemic variants of the virus with high morbidity and mortality. We investigated the protective activity of Ingavirin against the lethal influenza A (H1N1) 2009 virus infection on albino mice. Oral use of Ingavirin resulted in sharp decreasing of the mortality (index of protection up to 57%), slight decreasing of the infectious titer of the virus in the lungs (up to 40-fold), normalizing of the body weight dynamics and the lung tissue structure vs. the placebo-treated control. The degree of the bronchial epithelium damage was also strongly decreased. The results allow to consider Ingavirin as an effective antiviral against the current pandemic influenza virus.

The Ingavirin antiviral properties with respect to the parainfluenza virus, as an actual human respiratory tract pathogen, were investigated by two methods, i.e. immunoenzymatic analysis and microtetrazolium test. The results showed that along with the immediate antiviral activity Ingavirin had nonspecific cytoprotective properties. While affecting the virus proteins synthesis, Ingavirin lowered the virus cytopathogenic action. The drug significantly decreased the portion of the bronchial epithelium cells killed at the stage of acute infection.

Antiviral properties of Ingavirin were investigated in the Hep-2 cell culture with respect to the human respiratory tract virus (type 5 adenovirus). In concentrations of Ingavirin of 1000, 100 and 10 mcg/ml the generated posterity showed lower infective capacity (by 250, 100 and 10 times respectively). The electron microscopy of the infected cells confirmed the Ingavirin ability to disturb the adenovirus normal morphogenesis.

The epitope presentation system for ectodomain of M2-protein of influenza A virus (M2e) based on Cowpea Mosaic Virus (CPMV) was constructed for expression in plants Vigna unguiculata. CPMV is widely used as a vector for production of immunogenic chimeric virus particles (CVPs) bearing epitopes of different infectious human and animal pathogens. To produce chimeric CPMV virus particles in plants, two binary vectors were constructed bearing modified gene coding for S-coat protein of CPMV with insertions of M2e epitopes of human influenza and bird influenza viruses. Antigenic and immunogenic properties of CVPs obtained were investigated in mice immunization experiments and it was shown that they can induce anti-M2e IgG production and partial protection mice against challenge with low doses of flu virus. However, low infectivity and immunogenicity of CPMV chimeric particles indicate the need for further optimization of plant virus-based systems for M2e-epitopes presentation to use plants as a possible source of flu vaccines.

Aim: To confirm and prove on the extended contingent of volunteers the non-reactogenicity, safety and immunogenicity of "Grifor" vaccine in comparative trial with registered in Russia commercial vaccine "Vaxigrip"., Materials and Methods: Phase II clinical trial was performed on the research bases of Mechnikov Institute of Vaccines and Sera and Institute of Influenza. In single-blind comparative prospective randomized trial 300 adult volunteers (150 volunteers on each base) aged 18 - 60 y.o. were divided on 3 equivalent groups. Assessment of antigenic characteristics of "Grifor" vaccine was performed using hemagglutination inhibition assay (HAI) with chicken erythrocytes measuring geometric mean titer (GMT), seroconversion factor as well as level of seroconversion and seroprotection., Results: Previously performed studies proved non-reactogenicity, safety and high immunogenicity of "Grifor", whereas this comparative trial performed with commercial vaccine "Vaxigrip" did not reveal significant advantage in any of studied vaccine., Conclusion: "Grifor" vaccine meet the requirements of both EMEA CPMP and methodic guidelines MY 3.3.2. 1758-03 for inactivated influenza vaccines, which allows to register vaccine "Grifor" in Russian Federation.

During the latter half of 2005 a widespread outbreak caused by influenza highly pathogenic H5N1 virus among wild and domestic birds occurred in Russia. As pathogenicity level is a polygenic feature and majority of individual genes of influenza A viruses contribute to pathogenicity of influenza viruses to birds, animals and humans. Nucleotide sequencing of the entire genome of influenza H5N1 virus isolates obtained in Kurgan region (Western Siberia) was performed. Structure of viral proteins was analyzed according to the predicted amino acid sequences. HA receptor-binding site of A/chicken/Kurgan/05/2005 and A/duck/Kurgan/08/2005 strains was typical for avian influenza viruses and contained Glu and Gly at positions 226 and 228, respectively. Structure of the cluster of positively charged amino acid residues at the cleavage site was identical for all isolates: QGERRRKKR. According to the data of neuraminidase structure analysis NA of the H5N1 isolates tested was suggested to belong to Z genotype. Amino acid residues typical for birds were revealed in 30 out of 32 positions of M1, M2, NP, PA and PB2 proteins determining host range specificity. One strain isolated in Kurgan contained lysine in position 627 of PB2 protein. Kurgan isolates was shown to have remantadine-sensitive genotype. Glutamic acid was found at position 92 of NS1 protein in both strains indicating virus resistance to interferon. Phylogenetic analyses allowed relating Kurgan isolates to subclade II of clade II of highly pathogenic H5N1 influenza viruses.

The possible antiviral activity of preparations with antioxidant and/or antihypoxant properties was studied on two in vitro models of influenza infection: (i) in cultures of chorio-allantoic membranes of chicken embryos and (ii) in MDCK cells. Preparations under study were hypoxene, reduced glutathione, dihydroquercetin, trolox, coenzyme Q10, and the enzymatic preparation of superoxide-dismutase (recsod). Preparations possessing combined antioxidant/antihypoxic and detoxicating properties (reduced glutathione and hypoxene) produced a significant antiviral effect and enhanced the antiviral effect of rimantadine. The antiviral effect of these preparations was manifested by a decrease in the production of viral particles and, to a more pronounced degree, by the inhibition of cytopathogenic action of virus on cultured cells, which was revealed in the tests for the activity of respiratory enzymes. In contrast to the compounds containing thio or sulfo groups, the antioxidants of "direct action" (free radical scavengers) - coenzyme Q 10, trolox, quercetin and the enzymatic preparation recsod did not show any pronounced protective effect and in some cases even enhanced the production of viral particles and decreased the antiviral action of rimantadine.

The effect of some preparations with antioxidant and/or antihypoxic properties was studied under the conditions of cytotoxic action of the antiviral drug rimantadine in an MDCK cell culture. The preparations under study were hypoxene, reduced and oxidized glutathione, metadoxil, trolox (water-soluble analog of vitamin E), dihydroquercetin, coenzyme Q, and recsod (superoxide dismutase enzyme preparation). The protective drug action on the model of cytotoxicity in vitro was observed for the preparations possessing a complex of antioxidant/antihypoxic and detoxicant properties: hypoxene, metadoxil, and reduced glutathione. The protective effect of preparations did not correlate with their direct antiradical activity with respect to the stable free radical DPPH. Some compounds of phenolic nature (trolox, coenzyme Q) and recsod enhanced the harmful effect of rimantadine on the culture cells under the conditions studied. A possible explanation of this fact could be the conversion, under certain conditions, of the effect of phenolic compounds from antioxidant to pro-oxidant, which is confirmed by some literature data. The results do not allow antioxidant preparations to be recommended for routine application as cytoprotectors during toxic stresses of different nature. The protective activity of such preparations should be estimated separately for each particular xenobiotic.

Aims: To determine whether influenza can trigger heart attacks, we investigated the impact of influenza epidemics on autopsy-proven coronary deaths., Methods and Results: We studied weekly death due to acute myocardial infarction (AMI) and chronic ischaemic heart disease (IHD) in autopsies conducted in 1993 to 2000 in St Petersburg, Russia. We plotted the weekly acute respiratory disease (ARD) counts and influenza epidemics against AMI and chronic IHD deaths. There were 11,892 subjects dying of AMI and 23 000 subjects dying of chronic IHD. Median age was 75 for women and 65 for men. In every year, a peak of AMI and chronic IHD deaths were present and coincided with the influenza epidemic and peak ARD activity. A similar pattern was seen for each subgroup of men, women, subjects 50 years or older, and subjects 70 years or older. When comparing the average influenza epidemic weeks to average off-season weeks, the odds for AMI and chronic IHD death increased by 1.30 (95% confidence interval (CI): 1.08-1.56) and 1.10 (95% CI: 0.97-1.26), respectively., Conclusion: Influenza epidemics are associated with a rise in autopsy-confirmed coronary deaths. Influenza vaccination should be advocated for patients at high risk of developing cardiovascular events.

Background: Viruses are the most dangerous infectious contaminants of human donor blood and blood products. The purpose of the study was to investigate the virus-inactivating properties of fullerene suspension regarding influenza virus in allantoic fluid of chicken embryos., Methods: Influenza virus was propagated in chicken embryos, water suspension of C60 fullerene was added to the allantoic fluid. The fluid was light-irradiated at constant oxygen flow through the specimen, and the dynamics of virus titer was studied by virus titration in MDCK cells. The morphology of virions was studied by electron microscopy (EM)., Results: Dramatic drop of infectious titer (8 to 1 log10 EID50) of the virus was observed within 2h after start of irradiation. No change of the titers was observed in control specimens without fullerene, or light, or oxygen. EM study revealed numerous defects of virions' morphology (destruction of outer membrane) leading to the loss of infectious properties of the virus., Conclusions: Water-insoluble fullerenes may be considered as a prospective way for inactivation of enveloped viruses in biological materials including blood products.

The purpose of the study was to investigate the influence of cationic polymer structure on the formation of DNA-polycation complexes and their transfection activity. Primary, tertiary, and quaternary polyamines with molecular masses ranging from 8000 to 200,000 were investigated. DNA-cationic polymer interaction was characterized by low gradient viscometry, dynamic light scattering, circular dichroism, UV spectrometry, flow birefringence, DNA electrophoresis, and electron microscopy. Transfection activity of the complexes was evaluated by the expression of reporter gene (beta-galactosidase) and using synthetic FITC-labelled oligonucleotides. Complex formation was found to be dependent on the structure and molecular weight of the polymer and the ionic strength of the solution. Secondary DNA structure in complexes was not disrupted, and DNA was protected from protonation. Cell lines of different origin were used for testing of transfection activity of the complexes. The sensitivity of the cells to transfection was established to be highly dependent on the cell line. DNA-polycation complexes are non-toxic according to MTT. Polyallylamine, and polydimethylaminoethylmethacrylate were found to be the most promising polycations for gene delivery. Transfection efficacy of their complexes with DNA to T-98G cells reaches up to 90-100%. It was found that optimal molecular mass of polydimethylaminoethylmethacrylate is in the range of 8000-50,000 Da.

This paper analyzes the results of cell therapy carried out during multimodality treatment of 23 patients with trophic ulcers of the lower limbs because of varicosity and postthrombotic disease. Twenty-five patients were entered into the control group provided the modern wound dressings Suprasorb. It has been demonstrated that cell therapy applying fibroblasts, strain 11 00/14, noticeably upgrades the treatment efficacy. Ulcer healing was attained in all the patients of the basic group. The times of epithelialization averaged 1.5 week for varicose and 3.2 weeks for postthrombotic ulcers. In the control group, the healing of varicose trophic ulcers was recorded in 86% and of postthrombotic ulcers in 78% of patients. The times of healing averaged 3.6 and 3.9 months respectively, Cell therapy as part of multimodality treatment of trophic venous ulcers excels the modern wound dressings as regards the efficacy.

Symmetrical peptide GYDTQAIVENNESTEYG (WT, Wild Type) identical to 35-51 aminoacid residues of human alpha-lactalbumin (HLA) and peptide GYDTQTVVNNNGHTDYG (ID, IDeal symmetry) homologous to beta-domain of mammalian alpha-lactalbumins can form amyloid-like fibrils in conditions required for fibrillogenesis of HLA. The latter peptide can also form fibrils in deionized water. Fibrils formed by these peptides can cause forming of HLA amyloid-like aggregates in physiological conditions. These results provide an evidence for presence of amyloidogenic determinant in beta-domain of alpha-lactalbumin. Thus, symmetry in the primary structure may play the role in fibrillogenesis of proteins.

Article is dedicated to analytical investigation of the problem of current technologies in construction and manufacturing of anti-influenza vaccines. Epidemiological events in July-November 2005 in Russia (mainly in Siberia) and later in Ukraine showed that Health Care system was not ready for that turn over of epidemiological situation. It was completely the same situation in other countries. There are two general questions of a readiness in pre-pandemic situation: level of a diagnostic monitoring of epidemiological situation and preparedness to fast production of actual vaccine preparations. First task can be solved by immediate production of diagnostic sets for regional branches of National WHO Centers, and a second one depends on application of a novel approaches in construction of a anti-influenza vaccines. The construction of anti-influenza vaccines is based on genetic engineering (reverse genetics) and manipulation with plasmids carried out basic viral genes. Reassortation technology for preparation of hybrid viruses is going to the past by objective reasons. Advanced technologies are safety in laboratories and in manufacturing facilities. Moreover, genetic engineering in this field allows to planing the construction of vaccines bank, when the prognoses for actual viruses include more then two strains with different antigenic properties.

We generated several attenuated recombinant influenza A vectors expressing the Mycobacterium tuberculosis early secretory antigenic target (ESAT-6) protein. The ESAT-6 protein was recently identified as one of the most promising protective antigens for cell-mediated immunity. The obtained vectors appeared to be capable of inducing ESAT-6 specific Th1 immune response in mice after intranasal immunization. We found that double immunization with two influenza vectors of different subtypes provided a significant level of protection in mice, when applied as prophylactic vaccine, as well as substantial therapeutic effect in mice with pre-established tuberculosis infection. Moreover, we found a strong synergistic effect when vaccination with Flu/ESAT-6 vectors was combined with isoniazid treatment, resulting in a dramatic reduction of bacterial load in the lungs of infected mice.

The aim of the work was to study the effectiveness of using human embryo fibroblast culture in complex treatment of trophic ulcers of venous etiology in 23 patients with trophic ulcers of lower extremities. The cause of the appearance of ulcers was postthrombophlebitic disease in 16 patients and varicose disease in 7 patients. A control group consisted of 25 patients (postthrombophlebitic disease in 18 patients and varicose disease in 7 patients). The human embryo cell culture grown on the wound cover "Foliderm" was used at the stage of epithelization in the main group, while in the control group the modern alginate, collagen, hydrogel, polyurethane and hydrocolloid covers Suprosorb--Suprosorb A, Suprosorb C, Suprosorb G, Suprosorb F and Suprosorb H were used. Healing of the varicose trophic ulcers in the control group was achieved in 86% of patients, of post-thrombophlebitic--in 78% of patients. The average period of healing was 3.6 and 3.9 months respectively. Healing of trophic ulcers in the main group took place in 100% of patients. The average period of healing was 1.5 week for varicose and 3.2 weeks for postthrombophlebitic ulcers. The cell therapy was shown to be a highly effective method in treatment of venous trophic ulcers.

A new test system based on in vitro assessment of the cellular viability and/or metabolism is proposed, which offers an informative approach to the screening of antiviral drugs. Cytotoxic effects of the antiviral drugs rimantadine and polyrem were studied on the cultures of some mammalian cells. The results of short-term (2 h) exposures with the drugs tested were close to their LD50 values in mammals, which makes the proposed test system promising for the assessment of drug toxicity in vivo for the whole organism. A study of the state of cell metabolism after a long-term (48 h) exposure showed that the system of endocytosis is more sensitive than other indices with respect to antiviral drugs. Is was demonstrated on the cell level that the binding of drugs into polymeric complexes can decrease the degree of its cytotoxicity: the toxicity of polyrem (a polymeric complex of rimantadine) was lower as compared to that of the equimolar concentration of rimantadine or a mixture of rimantadine with a polymeric carrier.

In this review of literature modern notions on the role of birds in the evolution of the pathogenicity signs and immune system of the main and intermediate hosts of influenza viruses, as well as on the mechanisms of overcoming interspecific barriers, are analyzed. The chronology of the spread of "avian" influenza among humans, starting from 1997, the properties of the natural reservoir of this infection, and in particular influenza viruses A, the ways of their variability and evolution are presented. The conclusion has been made that the mixing, joint evolution, recombination and reassortment of viral genomes may be caused by global events in individual geographical regions.

The paper contains an analysis of research on designing drugs based on acridine derivatives. The discussed series of compounds is of essential value since acridines belong to the group of natural compounds with the pronounced antibacterial and anti-tumor activity. Improved chemical-synthesis techniques made it possible to synthesize both simple and complex compounds of the acridine series; they displayed a clear pharmacological activity as anti-proliferative, anti-tumor and antiparasitic preparations. The ability to induce interferons (INF), type 1, is an expected property of simple acridine derivatives. A variety of INF inducers, now used clinically, have been designed recently on the basis of the above compounds. The most well-known acridine derivatives, their pharmacological properties, action mechanisms and outlooks for practical application are described in the paper. The unique qualities of acridines are primarily attractive due to the possibility of using them for the purpose-oriented designing of drugs. Thus, acridines were used as a basis to create the specific regulatory HIV-1 elements, proliferation inhibitors of leukemia cells and new anti-tumor drugs. The elaboration of complexes of acridines derivatives combined with peptides intercalating specifically into the DNA big or small grooves is the most outstanding trend of acridines' research--it opens up prospects for using them in the synthesis of compounds regulating the gene expression.

We studied the effect of influenza virus proteins--hemagglutinin, neuraminidase, nucleoprotein, and membrane protein--on hemostasis in vitro and in vivo. The obtained data demonstrated that the envelope proteins hemagglutinin and neuraminidase increased the plasma fibrinolytic and anticoagulant activities and the activity of human tissue plasminogen activator. Among the core proteins of influenza virus, membrane protein proved to have the highest activity; in contrast to hemagglutinin and neuraminidase, it inhibited fibrinolysis, increased the coagulation activity of the plasma, and decreased the activity of human tissue plasminogen activator. Combined action of hemagglutinin and neuraminidase increased the plasma fibrinolytic and anticoagulant activities exceeding their individual effects. Combined action of an envelope protein hemagglutinin and membrane protein also increased the plasma fibrinolytic and anticoagulant activities although to a lesser extent as compared to hemagglutinin alone. The obtained data indicate that the viral proteins are physiologically active and can induce hemostatic changes specific for influenza.

New water-soluble antiviral chemical agents, containing 10 to 30% of adamantane derivatives (amino-, aminopropyl-adamantane-, aminomethyl- and rimantadine), which were conjugated with polycarboxylic matrixes of the divinyl ether and maleic anhydride copolymers (DIVEMA), were developed. The polymeric drugs exhibited a low cytotoxicity (4 to 10 times less than rimantadine) and a wide spectrum of antiviral activity against influenza viruses, including both the remantadine-resistant strains of A/PR/8/34 (H1N1) and the B/Saint-Petersburg strain/71/77 as well as against herpes viruses of type 1, parainfluenza viruses of types 1 and 3 and RS-virus. A reduction of the viral infection titer in their reproduction in sensitive cells' cultures was more than 2.0 Ig ID50. Complete inhibition of viral-specific syntheses, registered by immune-enzyme assay (IEA) and by hemagglutination test was observed at low infection doses ranging from 1 to 100 ID50. The efficiency of the antiviral effect depends on a drug's molecular weight and a structure of chemical bonds between the adamantane nucleus and the polymeric matrix.

Special attention should be given to the safety of drugs prepared from animal products. Industrial production of cytomedines and cytamines includes treatment of organs and tissues from young animals. Studies of preparations by methods recommended by the World Health Organization indicate that this technology guarantees the absence of infectious agents, protooncogenes, nucleic acids, and prion proteins.

The present review deals with the analysis of lymphokine response in two main respiratory viral infections: influenza and respiratory syncytial (RS) infection. Immune response in these two diseases has great phenomenological differences. In particular, in RS infection the intensive response of macrophages and epithelial cells, accompanied mainly by the synthesis and secretion of tumor necrosis factor and interleukin 8, is observed. Due to this fact no protective immunity is formed in RS infection, in contrast to influenza. The specific features of immune and lymphokine response in RS infection make it difficult to develop protective vaccines. At the same time it is with RS infection that the development of chronic bronchopulmonary diseases is linked. The analysis of the role of lymphokine response in respiratory viral infections, particularly in influenza and RS infections, confirms the necessity of revising therapy both in the acute and subacute stages of these diseases.

The capacity of water-soluble complexes of fullerene C60-polyvinylpyrrolidone to inhibit the replication of influenza viruses was studied. In contrast to remantadine, these complexes inhibit the replication of both A and B viruses (including the remantadine-resistant strains). The complexes inhibit influenza virus replication at all stages of replication cycle.

Reviews reports on the creation of liposomal vaccines published during recent 25 years. Describes methods of antigen and immunomodulator incorporation in liposomes and their immunogenic properties. Presents data on development of influenza vaccines and prospects for their utilization as universal carriers of viral and bacterial antigens.

Sera from 926 patients were analyzed by PCR using universal primers to surface gene of hepatitis B virus (HBV). HBV DNA was detected in 195 specimens. There were no serological markers of HBV in 8.2% of these sera, but later they were detected in patients' sera. In patients without HBV DNA in the serum, specific HBV antigens and antibodies were detected in 62%. Only in 14% of them the clinical picture of the disease corresponded to acute viral hepatitis, although the blood for PCR analysis was collected during the late stages of the infection. The rest cases were referred to mixed hepatitis C + D. Comparison of the results of PCR test and detection of serological virus replication markers HBeAg and HBeAb showed the presence of HBV DNA in 28% cases without HBeAg.

The antigenic properties of 51 strains of influenza virus A(H1N1), isolated in different cities of Russia during the epidemic of 1998, were studied. Most of these strains (49) proved to be similar to virus A/Bern/07/95 in the antigenic structure of hemagglutinin, but 2 strains isolated in Ulan-Ude were found to be closely related to new antigenic variants of this virus: A/Beijing/262/95 and A/Fukuoka/c7/98. The analysis of the antigenic structure of influenza-like diseases (ILD) in different cities of Russia revealed that adenoviruses causing up to 10.9-14.6% of all acute respiratory virus infections dominated at the pre- and post-epidemic periods. RS-viruses, parainfluenza viruses of types 2 and 3 circulated during the whole season (their proportion was 5.1-6.6%). The intensity of the circulation of influenza viruses A(H1N1) and A(H3N2) increased, starting from January, and continued till April 1998; its peak was observed in February-March in most of the cities of Russia (up to 37.5-41.6% according to the results of immunofluorescent diagnostics and 53-73% of ILD according to the results of the hemagglutination inhibition test). The occurrence of influenza B during this season was very low.

We studied the effect of influenza virus hemagglutinin on the volume of erythrocytes and aggregation activity of thrombocytes. Hemagglutinin proved to increase the erythrocyte volume, which ends in their lysis, and to induce disaggregation of the thrombocytes. The effect of hemagglutinin on the erythrocytes and thrombocytes was dose-dependent.