[The development of experimental sample of immune chromatographic test-system for detecting protein of pathogenicity of CagA Helicobacter pylori.]

The immunochromatographic test-system was developed for detecting protein of pathogenicity of CagA Helicobacter pylori in various biological samples (feces, content of dento-gingival recesses) and also in culture of H.pylori. The test-system represents multi-membrane composite on the basis of membranes manufactured by "MDI" (India). The main immunochemical components of test-system are conjugate of nanoparticles of colloid gold with size of 30 nm with monoclonal antibodies (clone HP-387), applied to membrane for conjugate; monoclonal antibodies (clone HP-1811) and anti-species antibodies of goat against Ig of mouse, applied to nitrocellulose membrane correspondingly in test and control zones. All antibodies are produced by the firm "Bialeksa" (Russia).

[The comparative evaluation of components used in development of immune chromatographic test-systems].

The article summarizes data of research studies concerning searching optimal conditions of development of highly specific and highly sensitive immune chromatography test-systems designed for detection of agents of infectious diseases. The analysis was implemented concerning choosing of optimal size of nano-particles of colloid gold and concentration of antibodies for production of conjugate. The alternatives are presented concerning the most frequently used combinations of buffer solutions applied for development and analysis of immune chromatography test-systems. The preferences are established in the field of characteristics of membranes used in composition of multi-membrane composite of immune chromatographic test-systems.

[INTESTINAL FAILURE AND YERSINIA PSEUDOTUBERCULOSIS TRANSLOCATION IN THE DEVELOPMENTOF EXPERIMENTAL GENERALIZED INFECTION].

THE OBJECTIVE: To determine the value of intestinal failure and translocation of bacteria Y. pseudotuberculosis, and normal intestinal microbiota in the initiation and generalization of infection in experimental pseudotuberculosis in conventional white mice, as well as pathological manifestation of it as a response to the adhesion and colonization of the mucosus membrane by pathogenic bacteria Y. pseudotuberculosis. MATERIALS AND METHODS: Experimental models of pseudotuberculosis in conventional white mice used the pathogenic Y. pseudotuberculosis 147 serotype I strain, containing a calcium-dependence plasmid with a molecular weight of 47 MDa. Cultivation of the pseudotuberculosis pathogen given its psychrophilic was performed on Hottinger agar at a temperature of (4-5) °C. The lactobacilli strain L plantarum 8P-A3 was isolated from a lyophilized commercial probiotic Lactobacterin (manufactured by "NPO Microgen", Russia) and used to obtain native culture supernatant fluid of lactobacilli, the composition of which was detected by gas-liquid chromatography with mass-selective detection. Gentamicin for parenteral administration was manufactured by JSC "Biochemist", Russia. Pathomorphological examination was performed on the 4-6th day of the experiment. Fragments of the small intestine, liver, kidneys, and lungs from dead animals were chosen for examination. Tissues were fixed in 10% neutral formalin, dehydrated in isopropanol and embedded in paraffin. Preparations were stained with Ehrlich hematoxylin and eosin, examined on the microscope "Mikmed-2" (JSC "LOMO", Russia) under magnification x 200-x1000. Statistical processing of the experimental results was carried out according to the method of Kerber in modification of I.P. Ashmarin and A.A. Vorobyov. RESULTS: The role of intestinal failure and translocation of bacteria Y. pseudotuberculosis, and normal intestinal microbiota in the initiation and generalization of infection in animals has been found. It has been proved that the oral administration of supernatant fluid containing microbial metabolites to animals as intramuscular administration of gentamicin equally prevent the development of generalized pseudotuberculosis and intensity of pathomorphological changes in the intestine and other organs of animals. CONCLUSION: Metabolites of the probiotic lactobacilli strain L plantarum 8P-A3 jugulate the development of pseudotuberculosis at an early stage of the pathological process in experimental animals infected with pathogen Y pseudotuberculosis, not only causing the preservation of the colonization resistance of the intestinal mucosa that prevents the adhesion and colonization of the pathogen, but also through their antimicrobial impact on the dissiminated pseudotuberculosis patho-gen bacteria in animals.

[COMPARATIVE EXPERIMENTAL EVALUATION OF THE EFFECTIVENESS OF MODERN PROBIOTICS, PREBIOTICS, SYNBIOTICS AND METABIOTICS FOR CORRECTION OF INTESTINAL MICROBIOCENOSIS IN ANIMALS WITH ANTIBIOTIC-ASSOCIATED DYSBIOSIS].

Probiotics for decades were leading in the arsenal of tools for correction of intestinal microbiocenosis. However, the response of representatives of intestinal microbiota to enteral intake of probiotics is variable, and in some individuals may be missing. Despite the fact that scientists still disagree on the necessity of forced settlement of normal microbiota at intestinal dysbiosis, large number of medicinal products, functional foods, dietary supplements, which efficiency is not always straightforward, is available for sale. The results of comparative experimental study of the effectiveness of 18 modern preparations for correction of intestinal microbiocenosis at antibiotic-associated dysbiosis in animals are presented. Conclusion is done on the need to evaluate efficacy and safety of tools developed for intestinal microbiota correction in animal experiments, followed by confirmation of clinical trials.

[Identification of genes encoding ligninolytic enzymes in naturally occurring basidiomycete isolates].

The presence of genes encoding lignin peroxidase, laccase, and manganese peroxidase was assessed in more than 20 types of polypore fungi collected in the woods of Kirov oblast; the fungi studied had not been previously characterized with regard to ligninolytic enzyme production. Fifteen isolates of eleven basidiomycete species were shown to contain genes coding for all three ligninolytic enzymes. Genes coding for these enzymes were detected in D. mollis, D. quercina, F. pinicola, G. trabeum, G. lucidum, H. fasciculare, L. betulina, P. betulinus, P. igniarus, P. pomaceus, P. pini, and P. cinnabarinus for the first time.

[The experimental evaluation with flow cytofluorimetry technique of the level of cellular immunologic memory in persons vaccinated against plague and anthrax].

The article deals with experimental evaluation with flow cytofluorimetry technique of the level of cellular immunologic memory in persons vaccinated with plague and anthrax live dry vaccines. It is established that the introduction of plague and anthrax live dry vaccines into organism of vaccinated persons ignites immunologic rearrangement manifested by reliable increase of level of blood concentration of Th1-lymphocytes (immunologic memory cells) against the background of vaccination. The higher correlation coefficient is detected between leucocytes lysis coefficient and stimulation coefficient according blood concentration level of T-lymphocytes predominantly at the expense of Th1-lymphocytes. The values of stimulation coefficient were calculated for corresponding blood cells of vaccinated persons. This data testifies the effectiveness of application of vaccination against plague and anthrax.

[Current laboratory methods to evaluate the efficiency of immunization against dangerous and extremely dangerous infections].

By taking into account the leading role of specific cellular immunity in the development of protection against the majority of dangerous and extremely dangerous infections of bacterial, viral, and rickettsial nature, the study of the above immunity should be considered as most important on estimating the body's immunological rearrangement when vaccines against respective infections are administered. The battery of the test used to date for the analysis of specific cellular immunity is mainly restricted to skin tests that have disadvantages. This paper reviews the currently available laboratory methods for in vitro studies of a specific cellular immune response as a criterion for evaluating the efficiency of immunization.

[Survival of probiotic microorganisms in the conditions in vitro imitating the process of human digestion].

PURPOSE OF THE STUDY: Assessment of survival bifidobacteria and lactobacteria under the conditions in vitro, simulating digestion in human stomach and intestine, and study of survival probiotic and indigenous microorganisms in co-cultivation on solid nutrient medium. MATERIALS AND METHODS: Probiotic microorganisms from commercial preparations Bifidobacterin and Lactobacterin, clinical isolates lactobacillus (Lactobacillus acidophilus No 1, L. brevis No 2) were used in experiments. Survival study of probiotic microorganisms was performed on a model in vitro, simulating the process of digestion in the human body. Assessment of the relationship of probiotic microorganisms and indigenous microorganisms was carried out in co-cultivation in vitro on solid nutrient medium. RESULTS: A significant reduction in the number of viable probiotic microorganisms during their incubation in model media was set as well as suppression of probiotic microorganisms growth by cultures of a clinical strains of lactobacillus, corresponding to biocompatibility by type "host against probiotic". CONCLUSION: While choosing probiotics in the treatment of dysbacterioses the character of relationship between probiotic microorganisms and indigenous microorganisms of a patient is recommended to be preliminarily tested. Also microorganisms of own microflora should be stimulated using modern prebiotics.

[The development and testing of a molecular biological test system for DNA detection of anthrax pathogen by real-time polymerase chain reaction assay].

The paper presents the results of the development and testing of a molecular biological test system for DNA detection of anthrax pathogen (Bacillus anthracis) by real-time polymerase chain reaction assay. The test system has shown high sensitivity, specificity, and reproducibility of results of analysis, as exemplified by aqueous suspensions of daily agar cultures of Bacillus anthracis strains, related and heterologous species of microorganisms, and clinical materials of experimental animals. There is evidence for the persistence of the basic characteristics of the test system when stored at 22 +/- 2 degrees C for 12 months.

[Comparative assessment of survival of probiotic microorganisms from commercial preparations under the conditions in vitro].

PURPOSE OF THE STUDY: Assessment of survival of probiotic microorganisms from commercial preparations under the conditions in vitro simulates digestion in human stomach and intestine. MATERIALS AND METHODS: Probiotic microorganisms from domestic and foreign commercial preparations were used in experiments. Study of survival of probiotic microorganisms was carried out on the model in vitro simulates the conditions of digestion in the human body. RESULTS: A significant reduction in the number of viable probiotic microorganisms in the 20 of 23 studied commercial preparations during their incubation in the model media is established. Probiotic microorganisms in the preparations Bactisubtil, Sporobacterin and Biosporin possess a good survival under the conditions in vitro. CONCLUSION: The selection of probiotics for the correction of intestinal microbiocenosis requires an individual approach that takes into account the character of relations of probiotic microorganisms and indigenous microorganisms of patient as well as the possibility of using advanced prebiotics in the combine treatment of dysbacteriosis.

[Experimental study of sensitivity of probiotic microorganisms to antibacterial drugs].

PURPOSE OF THE STUDY: Investigation of sensitivity of probiotic microorganisms to antibacterial drugs. MATERIALS AND METHODS: Probiotic microorganisms from domestic and foreign commercial preparations were used in experiments. The study of sensitivity of probiotic microorganisms to antibacterial drugs was carried out using solid nutrient medium containing calculated amount of antibacterial drugs. RESULTS: Probiotic microorganisms are mainly sensitive to the representatives of the major classes of antibacterial drugs. Only some of the studied microorganisms are resistant to 1-4 antibiotics. CONCLUSION: When choosing a probiotic in the treatment of dysbacteriosis need to consider whether th antibiotic therapy was carried out. The use of antimicrobial drugs, even in therapeutic doses, reduces the viability of probiotic microorganisms and their survival in the gut.

[Genotyping of the Bacillus anthracis vaccine strains using the multiloci VNTR-analysis].

The genotyping variety of 5 known anthracis vaccine strains using 18 variable loci of the chromosomal localization taken from a microbe culture collection of 48 Research Institute of Ministry of Defense was revealed in the research. The stability of the VNTR-loci was shown to be inherited from the B. anthracis strains with common origin and an opportunity of their gene-identification application. The gene profile of each analyzed vaccine strain using every 18 polymorphic loci was determined and the amplification products were sequenced. The variation of electrophoretic mobility of the amplifiers was found to be caused by the presence of the replication elements with various numbers of copies in their structure.

[The time course of changes in cell immunological parameters during administration of live dry plague vaccine].

The study of the time course of changes in cell immunological parameters by a magnetic separation technique in human beings during the administration of plague vaccine in relation to the immunological load revealed the higher blood levels of all T lymphocyte subpopulations on day 14 after vaccination. These changes are most typical of a primary vaccinated cohort. The increased frequency of plague vaccine administration and multiple immunizations with live plague, anthrax, and tularemia vaccines produce the time-course of changes in T lymphocyte populations (subpopulations) in response to the regular administration of plague vaccine. A high immunological load in man also promotes a significant reduction in the level of B lymphocytes.

[New immunoglobulin for treatment of anthrax].

AIM: To experimentally assess activity and safety of anti-anthrax intravenous immunoglobulin manufactured on standard technology. MATERIALS AND METHODS: Plasma from selected donors vaccinated with combined anthrax vaccine was tested by enzyme immunoassay. Samples of plasma with increased titer of anti-anthrax antibodies were merged in one manufacturing load and fractionated in ethanol at negative temperature according to standard technology. Formulation of intravenous immunoglobulin was manufactured according to standard technology of acid-enzyme hydrolysis. RESULTS: Proved medical technology of donors immune plasma fractionation provided 4 - 8-fold concentration of anti-anthrax antibodies. The finished product contained 5% of protein and was apyrogenic, non-toxic, thermostable, electrophoretically homogenous, had pH 6.65 and meet the requirements for manufacturing batches of human intravenous immunoglobulin. CONCLUSION: Protective effects of experimental human anti-anthrax immunoglobulin were comparable with control biological--equine anti-anthrax immunoglobulin for intramuscular use.

[Temperate Legionella bacteriophage: discovery and characteristics].

For the first time, temperate Legionella bacteriophage was isolated from organs of guinea pig infected with Philadelphia 1 strain of Legionella pneumophila. Negative colonies of bactriophage from 1.5 to 2.5 mm in diameter were detected. Central part of them was transparent and surrounded by peripheral zone of partial lysis. Electron microscopy showed that corpuscles of the phage consist from multifaceted elongated head of stretched hexagonal form and short tail. The bacteriophage lyzed bacteria, which cause Legionnaires' disease, and also had certain lytic activity against causative agent of tularemia.

[Identification of clinical isolates of Burkholderia cepacia and assessment of their pathogenicity in animal experiments].

In experiments on animals study of pathogenicity of 9 clinical strains of Burkholderia cepacia isolated from patients with chronic lung diseases was performed. Preliminary identification of studied strains by means of biochemical and genetic methods allowed to establish their belonging to B. cepacia species. It was determined that 6 of 9 strains are epidemiologically significant. Experiments showed that bacteria of studied strains are not able to cause infectious process in white mice and hamadryas baboons. Conclusion about appropriateness of development and use of other biological models was made.

[Assessment of immunity and allergy after vaccination with dry combined anthrax vaccine].

Study of humoral immune response and allergy in recipients of dry combined anthrax vaccine was performed. Immune response was assessed by antibody titers to protective antigen and by index of preventive properties of blood serum (PPS) of recipients. Relation of index of PPS and antibody titers in blood serum of the donors was established. Distribution of erythrocyte antigens in recipients of live dry and combined anthraxvaccines depending on blood group, Rh-factor, and age was studied. It has been shown that 80% of recipients of dry combined anthrax vaccine formed potent immunity with its high level lasted for 8 months. Study of allergenic properties of the combined anthrax vaccine using registration of neutrophils chemiluminescence in vivo showed low level of sensitization of vacinees.

[Treatment of late stages of anthracic infection].

Ways for increasing the anthrax treatment efficacy by combined use of antibiotics, toxin neutralizing, immunomodulating, symptomatic and pathogenetic agents are considered. The dynamics of the infection development was studied on rabbits by the microbiological, cytochemical, biochemical and cytomorphologic criteria. The efficacy of monoantibacterial and complex schemes of the etiopathogenetic therapy of the disease at the early and late stages (phases) of the infection generalization was estimated.