Study of Antiviral Activity of Metabolites of a New Serratia species K-57 Strain.

The results of studies of a newly isolated Serratia species K-57 strain are presented. The strain is characterized by antiviral activity towards human influenza A/Aichi/2/68/H3N2, vaccinia, mouse smallpox, and herpes simplex-2 viruses. The detected characteristics of the strain, including the data on activities on nucleolytic enzymes, recommend it for the development of therapeutic and preventive antiviral drugs.

Evaluation of the Effectiveness of Metabolites of Bacterial Strains Bacillus thuringiensis against Human Influenza Virus A/Aichi/2/68 (H3N2) In Vitro and In Vivo.

The morphological and physiological characteristics of Bacillus thuringiensis strains were analyzed and conditions for obtaining culture fluid with maximum yield of secreted RNases were determined. Zymographic analysis showed that culture fluid of B. thuringiensis strains along with low-molecular-weight (15-20 kDa) RNases contained enzymes with a molecular weight ~55 kDa and their content depended on the duration and conditions of culturing. Preparations based on B. thuringiensis culture fluid were effective against human influenza virus A/Aichi/2/68 (H3N2). In experiments on mice infected with 10 LD50 influenza virus strain A/Aichi/2/68 (H3N2), we selected effective variants of preparations based on culture fluid of B. thuringiensi strains for preventive administration that provided reliable protection of infected animals (protection coefficient 50%), close to that of the reference drug Tamiflu.

[MODERN METHODS OF PHYSICAL-CHEMICAL RESEARCH IN GASTROENTEROLOGY PRACTICE: THE EXPERIENCE OF INTERACTION].

UNLABELLED: The aim of this work was to assess the potential of some physical and chemical methods for studying erythrocytes and blood serum in gastroenterological practice by the example of colorectal cancer (CC). MATERIALS AND METHODS: A total of 26 persons with various stages of colorectal cancer and 16 healthy (control group) were examined. Parameters of erythrocytes and blood serum were investigated by light microscopy, dielectrophoresis in a non-uniform alternating electric field (DEF in NUAEF), terahertz spectroscopy, ellipsometry, Raman-spectroscopy. RESULTS: Polymorphism of erythrocytes, rigidity, viscosity, indexes of aggregation and destruction were significantly higher in patients with CC and polarizability, amplitude of erythrocyte deformation in NUAEF being lower than those in the controls. The study of erythrocytes by terahertz spectroscopy revealed the low levels of amplitude transmittance over the whole frequency range in CC patients compared to the controls. The increasing of refractive index, degree of heterogeneity of thin films obtained from the serum in CC patients were observed in considering the ellipsometric parameters. We found a significant increasing of the concentration of antigens to CD24 at the early stage of the disease. The areas of some peaks in Raman spectra were significantly lower in patients with CC compared to the healthy ones, it is possible due to a carotin deficiency. Most of the studied parameters were correlated with the stage of the disease. A set of optical methods for studying blood serum compared with those of histology and radiological methods of diagnosis showed their high sensitivity and specificity, positive and negative predictive value (80 % and above). CONCLUSION: The obtained results of the pilot study demonstrate the prospects of using physical and chemical methods of research of erythrocytes and blood serum for early diagnosis, stage of disease and monitoring the effectiveness of treatment of CC.

[Validation of the modified algorithm for predicting host susceptibility to viruses taking into account susceptibility parameters of primary target cell cultures and natural immunity factors].

The paper presents results of testing a modified algorithm for predicting virus ID50 values in a host of interest by extrapolation from a model host taking into account immune neutralizing factors and thermal inactivation of the virus. The method was tested for A/Aichi/2/68 influenza virus in SPF Wistar rats, SPF CD-1 mice and conventional ICR mice. Each species was used as a host of interest while the other two served as model hosts. Primary lung and trachea cells and secretory factors of the rats' airway epithelium were used to measure parameters needed for the purpose of prediction. Predicted ID50 values were not significantly different (p = 0.05) from those experimentally measured in vivo. The study was supported by ISTC/DARPA Agreement 450p.

ADAPTATION OF THE CORN EARWORM SINGLE NUCLEOCAPCIDE NUCLEOPOLYHEDROVIRUS (HELICOVERPA ZEA SNPV) FOR THE CONTROL OF THE COTTON BOLLWORM (HELICOVERPA ARMIGERA) POPULATION.

Helicoverpa zea (Boddie, 1850) (Hz) single nucleocapcide nucleopolyhedrovirus (SNPV) was adapted to the cotton bollworm (Helicoverpa armigera, (Hübner, 1805) (Ha)) by five blind passages on larvae. The full genomic sequence of the resulting strain HS-18 has been determined (GenBank acc. №: KJ004000.1). Biological activity of the HS-18 strain is higher than the activity of all other Russian strains of NPV, as far as cotton bollworm strain HearSNPV-G4. HS-18-infected caterpillars at the 3-rd and 4-th ages died much faster than those infected with HearSNPV-G4 strain. A major difference of HS-18 genome is an 18 bp repeat in the RING-finger ORF that confirms high variability of this region. Three other insertions and seven base substitutions were observed earlier, while six base substitutions are new. Mutations are located at ORF42, lef-9, ORF58, VP39, PIF-4, P48, SOD, ORF111, ORF129 and ORF138 genes. Among all nucleotide mutation only one is synonymous. Thus we suppose the selective pressure to the virus. The resulting strain HS-18 is recommended as a biopesticide for controlling the number of cotton bollworm in cotton fields.

[Highly pathogenic avian influenza: a new pandemic threat and possibilities to resist it].

The review presents modern data on outbreaks of avian influenza subtype H5N1 and the incidence of the infection in animals, birds, and humans. The authors adduce data on the resistance of influenza A virus to physical and chemical factors. Factors making avian influenza A virus potentially capable of causing human pandemia are considered. The review also deals with public health possibilities in the event of influenza pandemia, and measures directed towards lowering its negative social and economic consequences.

[Clinical trials of oral recombinant bivaccine against variola and hepatitis B during double vaccination].

Clinical trials of oral live recombinant embryonic variola and hepatitis B bivaccine as tablets (Revax-BT) were performed. When volunteers were prevaccinated with oral variola vaccine first in a small dose and, 7, 14, 30, 90, and 180 days later, in a larger dose, a slight reactoginicity was sometimes observed after the first vaccination (with a small dose) whereas revaccination with a larger dose did not give rise to any clinical manifestations. A month after vaccination, a protective level of virus-neutralizing antibodies to vaccinia virus (VV) was observed in 90-100% of the volunteers twice immunized with the bivaccine (in a small dose and in a larger one at an administration intervals of 1-2 weeks under remote revaccination while 6-9 months following vaccination, this level was recorded in 80% of the volunteers. A month following vaccination, 50-55% seroconversion to VV was observed in the volunteers twice immunized with the bivaccine (at an interval of 1 or 3-6 months). Cellular immunity to VV was low (0-20%). Double immunization of volunteers with the oral bivaccine under remote vaccination failed to produce the significant levels of humoral and cellular immune responses to hepatitis B markers. Recombinant VV was not recorded in any blood, saliva, and urine samples taken in the volunteers twice immunized with the bivaccine.

A prototype prophylactic anti-influenza preparation in aerosol form on the basis of Abies sibirica polyprenols.

This research investigates a promising antiviral compound based on polyprenols from Siberian silver fir (Abies sibirica). The physico-chemical characteristics of a preparation developed in aerosol form and an estimation of its protective efficacy against aerosol challenge of laboratory animals are presented. It is shown that (1) by using a simple ultrasonic disperser one can obtain aerosol of three formulations studied with about 70% of its mass accumulated in the size range below 1.8 microm; (2) 40-100% of aerosol particles contain preparation for different formulations; (3) after delivering under specified schedules, the preparations as developed can protect up to 100% of mice against 5 LD(50) of influenza A/Aichi/2/68 (H3N2) virus aerosol infection. Animals inhaled twice the preparation doses (which were 100 times lower than injection ones of the same efficacy) and did not exceed 10 microg/mouse. It was shown that the mode of action of this immunomodulating preparation was nonspecific stimulation of immune cells' various activities.

Effect of intramuscularly injected polyprenols on influenza virus infection in mice.

This study demonstrates the possibility of achieving a prophylactic effect by intramuscular injection of Abies sibirica polyprenols for the control of influenza virus infection in mice. One of the five polyprenol preparations tested, preparation N1, which had the lowest hydrophilic-lipophilic balance (8.6), produced a significant protective effect when injected in a dose of 2000 microg/mouse 2 days before aerosol infection of mice with influenza virus. A moderate protective effect was also observed using a second preparation, designated N2. One day after aerosol infection, animals pre-treated with 2000 microg doses of the polyprenol preparations or Hanks' solution showed no difference in the level of interferon accumulation in the lungs. Three days after injection of preparation N2 and N1, a significant decrease in spleen and thymus weights was, observed in the mice. One day after injection of these preparations, the number of lymphocytes in the bronchoalveolar tract of the mice exceeded almost twice that seen in mice treated with placebo. After 3 days, relative and absolute numbers of macrophages decreased, whereas those of lymphocytes increased significantly. Three days after the administration of preparations N1 and N2, macrophages became approximately twice as active in absorbing zymozan granules. Preparation N1 affected the system of superoxide radical anion production to a greater extent than preparation N2. The production of radical anions by the macrophages of the bronchoalveolar tract in the mice, 1 day after intramuscular injection of preparation N1, was significantly higher than that seen on day 3 and that induced by preparation N2 1 and 3 days after injection. These data indicate that emulsions of polyprenols that have relatively low hydrophilic-lipophilic balance, inhibit influenza virus infection in mice through a modulation of the host immune response.

[Theoretical approaches to the evaluation of antiviral drug effectiveness]. / Nekotorye teoreticheskie podkhody k otsenke éffectivnosti protivovirusnykh preparatov.

It is suggested that (a) LD50 is the measure of the efficacy of antiviral drugs: (b) a disease is characterized by the "barrier" type of a pathological process when in pathogenesis there exists an event whose manifestation largely determines the outcome of a disease (recovery/severe form with a fatal outcome). If the event consists in the penetration of a pathogen into a susceptible cell and the consequent reproduction of an antigen, the value LD50 for man is expressed as the same parameter for a model animal and as the parameters determined by in vitro human and animal cell experiments. The parameters include the degree of viral production adsorption on the cell and the levels of its reproduction. The algorithms for evaluating the efficacy of viral drugs are based on the relationship. The adequacy of the relationship is illustrated by experimental findings.

[A new setup for the generation and studies of mono-disperse microbiological aerosols in medical-and-biological research]. / Novyi kompleks dlia polucheniia i izucheniia monodispersnykh mikrobiologicheskikh aérozolei v mediko-biologicheskikh issledovaniiakh.

A setup for the generation and studies of mono-disperse microbiological aerosols is described in the paper. Coefficients of 3 microm aerosol deposition in the respiratory tract of mice and rats were refined by using the above setup. The probability of deposition of such particles in the trachea and lungs of mice was proven to be equal to 1.2 +/- 0.1% and 2.6 +/- 0.2%, respectively. The probability for rats was equal to 3.2 +/- 0.2 and 11.8 +/- 0.9%, respectively. The distribution of deposited aerosol particles was determined by electron microscopy.

[The simple method of direct estimation of infectious process in mice and rats aerogenically infected with influenza virus]. / Prostoi metod priamoi otsenki infektsionnogo protsessa u myshei i krys, aerogenno zarazhennykh virusom grippa.

Multiplication of influenza virus in laboratory animals (mice and rats) after aerogenic inoculation was recorded directly (by the agent accumulation in the lungs and trachea) and indirectly (by interferon concentration in the lungs of mice). Thermal inactivation of influenza virus in chick embryo allantoic fluid was observed (by 4.5-6 Ig within 48 h at 37 degrees C). The authors claim that influenza (strain A/Aichi/2/68) infection in the respiratory tract of mice and rats can be experimentally validated by inoculation of chick embryos with 10 and 20% mouse or rat lung homogenate (undiluted or diluted 10-fold) or with 1 and 5% mouse and rat trachea homogenate, respectively, 48 h after aerogenic inoculation of animals, and the virus AID50 be thus determined.

[Prophylactic efficacy of aerosol preparations based on Abies siberica polyprenols in experimental influenza infection]. / Profilakticheskaia éffektivnost' aérozolei preparatov na osnove poliprenolov pikhty sibirskoi pri éksperimental'noi grippoznoi infektsii.

Preliminary investigations showed high preventive activity of two of three aerosol preparations of Abies sibirica polyprenols with nonionic surface active substances towards influenza infection. At least 2 aerosol administrations are needed to attain a high protective effect, the second dose depending on the first. Relationship between animal reaction to influenza virus infection changed in a nonmonotonous mode, depending on the drug dose injected during the first treatment: as the dose increased, the death rate first decreased and reached the minimum and then increased again. Such a reaction to aerosol treatment can be explained by the hypothesis of hyperstimulation followed by exhaustion of the host defense systems after high doses of the preparation.

[Mechanisms of action of aerosol preparations based on Abies siberica polyprenols in experimental influenza infection]. / Mekhanizmy deistviia aérozolei preparatov na osnove poliprenolov pikhty sibirskoi pri éksperimental'noi grippoznoi infektsii.

Humoral and cellular mechanisms of Abies sibirica polyprenol effects on nonspecific resistance of mice to influenza A/Aichi/2/68 virus were investigated. Two aerosol doses of polyprenols had a high protective effect in mice challenged with influenza virus. Aerosol polyprenol preparations in the studied doses induced no interferon or tumor necrosis factor production in the lungs. Lung macrophage counts and capacity to produce superoxide anion radicals increased in survivors after influenza in comparison with intact animals. Double aerosol administration of polyprenols prior to influenza infection promoted an increase in the thymus weight, bronchoalveolar tract cell counts (predominantly at the expense of lymphocytes), and of superoxide-producing potential of macrophages, which, in turn, can contribute to improvement of the defense potential of the organism towards influenza virus.

[Mechanisms of mice resistance to influenza virus A/AICHI/2/68 after preventive injection with polyprenols ]. / Mekhanizmy rezistentnosti myshei k virusu grippa A/AIChI/2/68 pri profilakticheskom vvedenii poliprenolov.

Preventive effect in influenza can be attained by intramuscular injections of fir (Abies) polyprenols. One of 5 tested polyprenol preparations (No. 1), injected 2 days before aerogenic infection with influenza virus, reliably protected mice from disease. Mice pretreated with polyprenol preparations or Hanks' solution did not differ by accumulation of interferon in the lungs One day after aerogenic infection. Three days after injection of polyprenol preparation No. 1 the weights of the spleen and thymus significantly decreased. One day after injection cell count in the bronchoalveolar tract of mice was almost 2-fold higher than in the control at the expense of lymphocytes and macrophages. After 3 days the relative and absolute counts of macrophages decreased and those of lymphocytes decreased significantly. Three days after injection macrophages were 2-fold more active in absorption of zymosan granules. Preparation No. 1 affected the production of superoxide anion radicals, whose production by all macrophages in the bronchoalveolar tract of mice was significantly higher on day 1 postinjection than on day 3 and higher than on days 1 and 3 after injection of preparation No. 2.

Infection of chickens caused by avian influenza virus A/H5N1 delivered by aerosol and other routes.

This study presents results of the study of infectivity of avian influenza virus (AIV) A subtype H5N1 strains isolated from agricultural birds across the territory of the Russian Federation and CIS countries. The results of the susceptibility of chickens to the AIV isolates delivered by the aerosol route and the dissemination of the virus in the organs of infected birds are presented. As was observed, the sensitivity of birds to AIV by the aerosol route of infection is 30 times higher than by intranasal route, 500 times higher than by the oral route and 10000 times higher than by the intragastric route of infection, which is indicative of higher permissivity of respiratory organs to AIV. The highest titres of AIV A subtype H5N1(A/Chicken/Kurgan/05/2005 strain) in aerosol-infected chickens were found in nasal cavity mucosa, lungs, cloaca, serum and kidney, where viable virus accumulation was detected by 18h post-infection (p.i.). The highest virus titres were observed 54h p.i. in lungs, serum and kidney, reaching the value of 8.16 lg EID50 /g(ml) in the lungs. The results showed that birds infected by the aerosol route developed higher titres of virus than those infected by other routes.

Inactivation and mineralization of aerosol deposited model pathogenic microorganisms over TiO2 and Pt/TiO2.

Air disinfection from bacteria and viruses by means of photocatalytic oxidation is investigated with microorganisms loaded over photocatalysts' films from aerosols. Deposition method and equipment have been developed to load Mycobacterium smegmatis , Bacillus thuringiensis , vaccinia virus, and influenza A (H3N2) virus on slides with undoped TiO(2) and platinized sulfated TiO(2) (Pt/TiO(2)). Inactivation dynamics was measured under UVA irradiation and in the dark. About 90% inactivation is reached in 30 min irradiation on TiO(2) and from 90 to 99.8% on Pt/TiO(2). The first-order inactivation rate coefficient ranged from 0.18 to 0.03 min(-1), over Pt/TiO(2) being higher than on TiO(2) for all microorganisms except Bacillus thuringiensis. The photocatalytic mineralization of Bacillus thuringiensis was performed on TiO(2) and Pt/TiO(2) with different photocatalyst and microorganism loadings. Completeness of mineralization depended on the TiO(2) to bacteria mass ratio. The rate of the photocatalytic carbon dioxide production grows with both the cell mass increase and the photocatalyst mass increase. Pt/TiO(2) showed increased rate of mineralization as well as of the inactivation likely due to a better charge carrier separation in the doped semiconductor photocatalyst. The results demonstrate that photocatalytic filters with deposited TiO(2) or Pt/TiO(2) are able to inactivate aerosol microorganisms and completely decompose them into inorganic products and Pt/TiO(2) provides higher disinfection and mineralization rates.

Inactivation of viruses in bubbling processes utilized for personal bioaerosol monitoring.

A new personal bioaerosol sampler has recently been developed and evaluated for sampling of viable airborne bacteria and fungi under controlled laboratory conditions and in the field. The operational principle of the device is based on the passage of air through porous medium immersed in liquid. This process leads to the formation of bubbles within the filter as the carrier gas passes through and thus provides effective mechanisms for aerosol removal. As demonstrated in previous studies, the culturability of sampled bacterium and fungi remained high for the entire 8-h sampling period. The present study is the first step of the evaluation of the new sampler for monitoring of viable airborne viruses. It focuses on the investigation of the inactivation rate of viruses in the bubbling process during 4 h of continuous operation. Four microbes were used in this study, influenza, measles, mumps, and vaccinia viruses. It was found that the use of distilled water as the collection fluid was associated with a relatively high decay rate. A significant improvement was achieved by utilizing virus maintenance fluid prepared by using Hank's solution with appropriate additives. The survival rates of the influenza, measles, and mumps viruses were increased by 1.4 log, 0.83 log, and 0.82 log, respectively, after the first hour of operation compared to bubbling through the sterile water. The same trend was observed throughout the entire 4-h experiment. There was no significant difference observed only for the robust vaccinia virus.