Vol 93, No 3 (2016)

Aim. Evaluation of antibacterial effect of y-globulin fraction bound and free copper and zinc cations, applied in cultures of S. aureus and P. aeruginosa at physiological (micromolar) concentrations. Materials and methods. Day cultures of S. aureus or P. aeruginosa were transferred from agar to physiological solution, and cell suspension was prepared, containing approximately 10³ - 10⁴ CFU/ml. Samples of metal-complexes of y-globulin with copper and zinc cations (30 and 45 pg/ml), control y-globulins (30 and 45 pg/ml) and salt solutions of copper and zinc, cation content in those corresponded to the quantity of the metal, that had bound with the protein at the stage of metal-complex obtaining (75 ng/ml), were introduced into the suspension. The suspensions were incubated at 37°C for 6 hours, sampling and CFU count according to the accepted micromethod was carried out every 2 hours. By the end of incubation (6 hours of observation) the suspensions were transferred into nutrient broth, thermostated for 1 day at 37°C, transparency of the nutrient broth compared with control (sterile) was evaluated afterwards. Results. Toxic effect of copper and zinc cations is detected starting from the 3rd hour of observation in S. aureus culture. Viable bacteria are absent in the culture with zinc after 6 hours, with copper - after 4 hours of incubation, y-globulin, that had bound copper cations, reduces the quantity of viable cells compared with control protein by 11.9 - 33.0% (p<0.05 - 0.1) at 4 and 6 hours of incubation. In P. aeruginosa culture, toxic effect of copper cations manifests immediately after initiation of the culture and results in realization of complete bactericidal effect after 4 hours of observation. Zinc cations do not have such properties, y-globulin, that had bound copper cations, reduces the quantity of viable cells compared with control protein at 4 and 6 hours of incubation by 19.3 - 25.8 % (p<0.001). Conclusion. S. aureus bacteria, supported in physiological solution are subject to toxic effect of physiological (micromolar) concentrations of free copper and zinc cations, and also copper cations, bound by human serum y-globulin. P. aeruginosa bacteria under the same conditions experience toxic effect of copper cations (but not zinc), free as well as bound by human serum y-globulin. Whereas a full bactericidal effect is realized in S. aureus and P. aeruginosa cultures in the presence of free cations of copper.

Aim. Typization of natural foci of HFRS of the Republic of Bashkortostan and differentiation of their territories by the degree of potential epidemic hazard. Materials and methods. Materials of epizootologic and epidemiologic monitoring of natural foci of HFRS of the Republic of Bashkortostan for 1980 - 2014 were used. Evaluation of area of territory with high, medium and low potential epidemic hazard by HFRS was carried out taking into account multi-year parameters of morbidity by administrative districts of the Republic of Bashkortostan. 3 gradations of morbidity level by HFRS - more than 201, 101 - 200, less than 100 per 100 thousand of the population - were used for epidemiologic differentiation of the territory. Results. 3 main landscape-epidemiologic types of HFRS foci were isolated - forest, forest-barrens, barrens, as well as their landscape variants. The area with a high degree of potential epidemic hazard by HFRS was established to be around 3.8%; medium - 15.9%, low - 80.3% of the whole territory of the Republic of Bashkortostan. Conclusion. A necessity of a differentiated approach to territories with various degree of potential epidemic hazard by HFRS is justified during execution of prophylaxis activities.

Aim. Comparative study of antigenic properties of recombinant proteins OspCgar and OspCafz and recombinant chimeric polypeptide OspCgar+afZ, that contains amino acid sequences of mature immune dominant OspC proteins of West-Siberian isolates of Borrelia garinii (OspCgar) and B. afzelii (OspCafz), and evaluation of possibility of their use as antigens during creation of test-systems for serodiagnostics of Lyme borreliosis (LB) on the territory of Western Siberia. Materials and methods. Recombinant chimeric polypeptide OspCgar+afz and recombinant mature proteins OspCgar and OspCafz, obtained by expression of the corresponding genes in Escherichia coli cells, purified by affinity chromatography in Ni-NTA-sepharose CL-6B and studied by EIA method for the ability to bind antibodies from sera of LB patients. Results. A difference in sensitivity of determination by EIA method of specific IgM and IgG against borreliae in blood sera of LB patients with localized stage of the disease during use of OspCgar, OspCafz and OspCgar+afZ chimera as antigens was shown. Chimeric antigen OspCgar+afz was established to show higher antigenic activity compared with each of the OspCgar or OspCafZ antigens separately. Conclusion. The results of the study allow to examine the recombinant chimeric polypeptide OspCgar+afz as a possible component during creation of test-systems for serodiagnostics of LB on the territory of West Siberia.

Aim. Study innate and adaptive immunity in patients with migrating erythema, clinical effectiveness of combined therapy using Immunovac vaccine and dynamics of immunologic parameters as a result of the therapy. Materials and methods. 37 adult patients with migrating erythema were examined. The patients were divided into 2 groups: 1st gr. (14 individuals) - Immunovac by intranasal-subcutaneous method against the background of basic therapy; 2nd gr. (23 individuals) - 200 mg/day doxycycline therapy for 21 days. Phagocytic activity of blood neutrophils; TLRs expression on mononuclear leukocytes of peripheral blood (PBML) and skin cells in foci by flow cytometry with mAT against TLR2, 3,4, 5,6, 7, 8,9 using flow cytometer FC-500; subpopulation composition of peripheral blood lymphocytes; levels of pro-, anti-inflammation and regulatory cytokines in blood sera by EIA method; IgG, IgM, IgA in blood sera were studied in patients before treatment and 1 month after therapy. Results. A high level of TLR2, 4, 7, 8 on skin cells in foci, TLR2, 4 - on blood cells; low content of CD95+ and CD25+, high level of serum IF-lb, IL-2 and IF-4, an increase of general IgE level was detected in patients with migrating erythema. Immunovac facilitated an increase of CD95+ and CD25+, IFN-y synthesis, reduced the level of general IgE in a more pronounced way than basic therapy. Conclusion. Inclusion of Immunovac into therapy of patients with migrating erythema facilitates increase of clinical effectiveness and correlates with correction of immunologic disorders.

Aim. Description of the first documented case of imported hepatitis E, associated with genotype 4 of HEV and introduced from southern France. Materials and methods. Clinical, epidemiologic and laboratory analysis of the imported case of disease of hepatitis E was carried out. Phylogenetic analysis of nucleotide sequences of HEV isolate, taken from the patient, was carried out. Results. Epidemiologic analysis allowed to assume imported character of the detected case of HEV-infection. Comparative analysis of nucleotide sequences of regions of the open reading frame 2 (300 nt) and open reading frame 1 (721 nt) of HEV genome, isolated from the patient, showed identity of this isolate with variants of genotype 4 HEV, isolated in France in 2009 - 2011 from patients with autochthonous hepatitis E. Conclusion. The results obtained confirm the case of import into Russia of genotype 4 HEV from south-eastern France (Corsica), where spread of this virus genotype is observed in recent years.

Aim. Analysis of results of epidemiologic monitoring of especially dangerous, natural-foci and other infectious diseases, as well as epizootologic activity of natural foci of infection on the territory of city-resort Sochi. Materials and methods. Laboratory studies of 820 samples by PCR, immune- and bacteriologic methods were carried out, among those 344 - clinical material, 12 - water from open bodies and 321 - field material. Molecular-genetic identification of 143 strains of Vibrio cholerae, isolated from open water bodies of city-resort Sochi, was carried out. Results. Circulation of causative agents of Q fever, tularemia and hemorrhagic fever with renal syndrome of Dobrava-Adler genotypes was established, as well as rickettsia of tick spotted fever group. Predomination of rotaviruses (70.9%) was detected during study of samples of clinical material in etiologic structure of sporadically emerging acute intestine infections. Relatively high temperature values of river water during summer months (from 18 to 30°C), exit of sulfide mineral waters into the riverbed, that create alkaline medium, seasonal reduction of river debit and speed of water current facilitated sustenance of contamination of water of Agura river by V. cholerae. Conclusion. Activity of natural foci of the infectious disease and contamination of Agura river by V. cholerae 01 (atoxigenic) gives evidence on the necessity of constant control of epidemiologic situation by dangerous and natural-foci infectious diseases on the territory of city-resort Sochi, as well as study regional features of their causative agents, including using genetic methods.

Pathogenicity factors of Corynebacterium non diphtheriae - pili, microcapsule, cell wall, pathogenicity enzymes, toxins, that determine the ability of microorganisms to consequentially interact with epithelium of entry gates of the organism, replicate in vivo, overcome cell and humoral mechanisms of protection, are examined in the review. Particular attention in the paper is given to species of non-diphtheria corynebacteria, that are pathogenic for human and able to produce toxins - Corynebacterium ulcerans and Corynebacterium pseudotuberculosis. Mechanisms of expression regulation of PLD-exotoxins, its interaction with immune system cells are described.

Five influenza pandemics emerged in the 20th and 21st centuries. Data regarding possibility of infection of swine with human influenza viruses and persistent circulation of these strains among swine with subsequent infection of humans with these viruses were obtained in the recent years. A possibility of prediction of influenza pandemics by constant observation and study of influenza viruses circulating among swine is discussed in the paper.