Dynamics of Changes in the cAMP/cGMP Concentration Ratio in the Thymus and Spleen of Laboratory Mice during Vaccination against Plague and Tularemia against the Background of Immunomodulation.

Vaccine strains Yersinia pestis EV NIIEG at a dose of 103 CFU and Francisella tularensis 15 NIIEG at a dose of 102 CFU induced changes in the concentration of cyclic nucleotides in the thymus and spleen of white mice. Antigen-induced changes in the cAMP/cGMP ratio in immunocompetent organs had a phase or oscillatory character, which seems to be related to the regulation of postvaccination immunoreactivity in the body. Synthetic organoselenium compound 974zh stimulated an increase in the amplitude of cAMP/cGMP oscillations, indicating its stimulating effect on the immunogenic properties of vaccine strains at doses an order of magnitude below the standard doses.

The Influence of the Synthetic Organoselenium Preparation 974zh on the Immunogenic Activity of Yersinia pestis EV Vaccine Strain NIIEG.

In the present study, a stimulating effect of a new synthetic organoselenium compound 2,6-dipyridinium-9-selenabicyclo[3.3.1]nonandibromide (974zh) on the immunogenic activity of the vaccine strain Yersinia pestis EV NIIEG was revealed. After infection with the virulent plague strain, the survival rate of laboratory mice immunized with the vaccine strain grown on Hottinger's agar in the presence of 974zh (300 µg/ml) increased in comparison with control animals immunized with the Y. pestis EV NIIEG culture grown on agar without the studied compound. Plasmid screening of cultures grown on medium with and without 974zh showed that plasmid DNA of Y. pestis EV culture grown in the presence of 974zh had broader bands in the control grown without 974zh. This phenomenon can indicate activation of replication of plasmid DNA of Y. pestis EV under the influence of the experimental compound.

Peculiarities of Humoral Immunity Formation in Medical Institution Employees Recovered from or Vaccinated against COVID-19 in Irkutsk.

We performed a seroepidemiological survey of the level and structure of population immunity to SARS-CoV-2 among employees of medical institution in the Irkutsk region during the COVID-19 pandemic. Seroprevalence assessment was organized from May 2020 to April 2021. The level of antibodies to SARS-CoV-2 was measured by ELISA. It was found that 139 (46%) of 299 examined workers were seropositive, including 50 (36%) vaccinated against COVID-19, 75 (54%) patients diagnosed with COVID-19, and 14 (10%) asymptomatic cases of SARS-CoV-2. The results obtained should be taken into account when predicting the dynamics of the epidemic process and organizing preventive (antiepidemic) measures, including vaccination.

Effect of a Synthetic Organoselenium Compound on Post-Vaccination Immunopoiesis in the Red Bone Marrow and Cell Composition of Peripheral Blood of White Mice Vaccinated with Yersinia pestis EV.

We studied the effect of an experimental synthetic organoselenium compound 2,6-dipyridinium- 9-selenabicyclo[3.3.1]nonane dibromide (974zh) on the cell composition of the red bone marrow and peripheral blood in white mice. The study drug co-administered with Yersinia pestis EV vaccine strain (103 CFU) potentiated maturation and migration of mature neutrophils from the bone marrow into the circulation. Reducing the dose of the live vaccine and the anti-inflammatory properties of the study drug made it possible to reduce the allergic reaction during the vaccination process.

Phylogenetic reconstruction of the initial stages of the spread of the SARS-CoV-2 virus in the Eurasian and American continents by analyzing genomic data.

Samples from complete genomes of SARS-CoV-2 isolated during the first wave (December 2019-July 2020) of the global COVID-19 pandemic from 21 countries (Asia, Europe, Middle East and America) around the world, were analyzed using the phylogenetic method with molecular clock dating. Results showed that the first cases of COVID-19 in the human population appeared in the period between July and November 2019 in China. The spread of the virus into other countries of the world began in the autumn of 2019. In mid-February 2020, the virus appeared in all the countries we analyzed. During this time, the global population of SARS-CoV-2 was characterized by low levels of the genetic polymorphism, making it difficult to accurately assess the pathways of infection. The rate of evolution of the coding region of the SARS-CoV-2 genome equal to 7.3 × 10-4 (5.95 × 10-4-8.68 × 10-4) nucleotide substitutions per site per year is comparable to those of other human RNA viruses (Measles morbillivirus, Rubella virus, Enterovirus C). SARS-CoV-2 was separated from its known close relative, the bat coronavirus RaTG13 of the genus Betacoronavirus, approximately 15-43 years ago (the end of the 20th century).

Antigen-Specific Activation of Blood Lymphocytes in Mice Immunized with Brucella abortus 19 BA.

We studied the expression of activation markers CD25 and CD69 by blood lymphocytes in white mice vaccinated with Brucella abortus 19 BA in antigen-specific tests in vitro. During incubation of blood lymphocytes with brucellosis polysaccharide-protein antigen, a statistically significant increase in the expression of CD25 by B cells and CD69 by T cells was observed; brucellin increased the expression of CD25 by B and T cells. Comparative analysis of the action of antigen preparations B. abortus showed that only brucellin has antigen-specific activity against CD19+CD25+ cells. The used method can be considered as a promising test for evaluation of the effectiveness of brucellosis immunoprophylaxis, which substantiates the need for further research.

Immunotropic Properties of an Experimental Synthetic Selenium-Organic Compound.

We studied immunotropic properties of synthetic selenium-organic preparation 2,6-dipyridinium-9-selenabicyclo[3.3.1]nonyl dibromide (974zh). The experimental preparation reduced the cAMP/cGMP ratio, which indicated an increase in proliferative activity of cells of immunocompetent organs (thymus and spleen) in experimental animals. It was shown that 974zh intensified the immune response to Yersinia pestis EV thereby increasing the resistance to the plague agent.

Effect of Synthetic Organoselenium Drug on the Degree of Pathological Changes in the Organs of White Mice Immunized with Tularemia and Brucellosis Vaccines.

We studied the effect of the organoselenium compound 2,6-dipyridinium-9-selenium-bicyclo[ 3,3,1]nonan dibromide (974zh) on the severity of pathological changes in the organs of experimental animals immunized with live tularemia and brucellosis vaccines. It was found that 974zh reduced reactogenicity of vaccines for experimental animals. Our findings indicate the prospects for further studies of the effects of 974zh on the functional state of experimental animals.

Effects of Thermoextracts of Brucella S and L Forms on Lipid Peroxidation and Antioxidant Defense in Organs of Laboratory Animals.

The dynamics of LPO marker malondialdehyde formation and peroxidase-destroying activity was studied in homogenized organs of guinea pigs, immunized with thermoextracts from S and L forms Brucella abortus I-206. The L form brucella thermoextract exhibited a lower reactogenicity and adequately activated the antioxidant system, due to which the destructive effects of ROS could be partially neutralized during the vaccinal process.

Effect of Metal-Containing Nanocomposites on Functional Status of the Thymus in Experimental Animals.

The effects of nanostructurized composites argentogalactomannan and argento-poly-1-vinyl-1,2,4-triasole on the kinetics of T cell maturation in the thymus of albino mice were studied. These nanocomposites stimulated the formation of CD3+CD4+CD8- and CD3+CD4-CD8+ cells from CD3-CD4-CD8- cells, the effect of argentogalactomannan was more potent. The efficiency of these compounds as agents improving the defense potential deserves special studies.

Immunogenesis in White Mice Infected with Yersinia pestis with Different Plasmid Composition.

Morphological changes in the immunocompetent organs of white mice with experimental plague infection manifested in activation of the immune response of different degree and pathological process of different severity that depended on the plasmid composition of Y. pestis. Widening of the T-dependent zones in the immune organs of white mice infected with isogenic strains of Y. pestis with different plasmid composition attests to activation of cellular immunity. Our findings allow considering Y. pestis subsp. altaica I-2948/3, Y. pestis subsp. pestis I-3479 and Y. pestis subsp. pestis I-3480 as promising candidates for vaccine strains.

[The loop mediated isothermal amplification of DNA: principle of method and perspectives of application in molecular diagnostic of cholera: publications review].

The article considers characteristics of technology of reaction of loop mediated isothermal amplification of DNA (LAMP), issues of optimization of reaction and perspectives of its application as a quick highly-specific test in molecular diagnostics of infectious diseases and monitoring of contamination of environment objects with pathogens. The analysis of publications data concerning application of LAMP in diagnostics of cholera testifies high diagnostic value. The LAMP supports possibility of direct rapid detection of toxin-producing strains of Vibrio cholerae in clinical samples. This technique also provides identification of determinants of cholera vibrio in pure culture, samples from environment objects and food products. The research studies established exceeding of parameters of sensitivity and specificity of LAMP as compared with polymerase chain reaction that permits considering LAMP as a perspective technique for express-analysis of clinical material from patients with suspicion on cholera. The LAMP technique can be also used in screening studies of environment objects. The development of test-systems based on application of this technology is required.

[ON SOME DEBATABLE PROBLEMS OF THE NATURAL NIDALITY OF PLAGUE].

The communication substantiates the opinion that the theory of natural nidality of plague; which is based on the fundamental recognition that fleas play a leading role in the transmission and accumulation of the plague pathogen, cannot be disproved or substantially changed on the alternative weakly reasoned assumptions and hypotheses. All its "bottlenecks" are quite understandable when considering the long-term volumetric materials that have been gathered directly in nature and generalized in multiple publications. Plague is an obligate transmissive infection; its, agent is a highly specialized parasite that is completely associated in its vital activity with the only group of the blood-sucking insects--fleas and that is transmitted through periodic colonization of warm-blooded animals for a short time. All other types of plague microbe persistence in nature are either occasional or minor and do not play any significant role in pathogen persistence in the natural foci of this disease. There are no strong grounds for seriously considering the attempts to revise the main points of the theory of natural nidality of plague, which are widely held in current academic publications.

[PLAGUE IN MANCHURIA (1910-1911) AND EBOLA VIRUS DISEASE IN WEST AFRICA (2014-2015): COMMON PREREQUISITES FOR THE DEVELOPMENT OF EPIDEMICS].

The paper gives the results of a comparative analysis of the prerequisites for the emergence and spread of epidemics of particularly dangerous infections, by using plague in Manchuria (1910-1911) and Ebola virus disease in West Africa (2014-2015) as examples. Analysis of literature and archival data and online information could reveal a number of common factors and conditions, which substantially contributed to the epidemics. Organization of anti-epidemic (preventive) measures in cases of the threatening epidemic spread, of particularly dangerous diseases must be based on the minimization, of the influence of the specific factors and conditions, which facilitate disease transmission in a given area in a given time.

[SPECIFIC FEATURES OF RELATIONSHIPS BETWEEN THE FRONTOPSYLLA LUCULENTA LUCULENTA (JORDAN ET ROTHSCHILD, 1923) FLEA AND THE PLAGUE PATHOGEN WITH DIFFERENT PLASMID COMPOSITION].

It was experimentally established that plague pathogen strains with different plasmid composition variously suppressed the viability of Frontopsylla luculenta luculenta fleas. Dead insects were most frequently observed among those infected with a virulent strain having the cryptic plasmid pTP33. The presence of the avirulent and apesticinogenic plasmid I-3480 in the fleas less deteriorated their state. Biofilm formation by different F.l.luculenta strains in the body was characterized by quantitative and qualitative differences. The strains that had the cryptic plasmid and were able to form the biofilm in the F.l.Iuculenta fleas surpassed the three-plasmid strain I-3230 and their formned aggregates achieved very large sizes and frequently persisted until the end of the experiment. Small solitary masses were generally observed in the insects infected with the three-plasmid strain. Thus, the pTP33 plasmid potentiated the pYT plasmid-encoded ability to colonize the F.l.Iuculenta fleas with the plague pathogen; in this case the products of the pYV and pYP plasmids (or one of them) are toxic to ectoparasites.

[Analysis of complete sequence of cryptic plasmid pTP33 from Yersinia pestis isolated in Tuva natural focus of plague].

This paper studies a full nucleotide sequence of cryptic plasmid pTP33, which was isolated from the typical plague strain of the Tuvinian natural focus, Yersinia pestis I-2638. Sequencing was carried out using the 454 GS Junior platform (Roche). In analysis using the software package GS De Novo Assembler v. 2.7 (Roche) and the algorithm Newbler v. 2.7, 1855 nucleotide reads, which contained 1101246 nucleotides, were assembled to a contig of 33 978 bp. The GC content of the obtained nucleotide sequence was 50.25%. During annotation, we found 56 open reading frames. Homologs of the predicted reading frames were sought in the BLAST databases. We detected 22 reading frames coding hypothetical proteins, 23 frames coding phagerelated proteins, and 11 frames coding proteins with known functions, including toxin­antitoxin system YefM-YoeB, nucleic acids and polysaccharides metabolism proteins (exopolysaccharide production protein ExoZ, exodeoxyribonuclease VIII), and replication proteins (ParA). Some predicted pTP33 proteins were found to be homologs (from 45 to 75%) with sequences of phage-related proteins of certain microorganisms­endosymbionts of insects (Sodalis glossinidius) and endosymbionts of entomopathogenic nematodes (Photorhabdus luminescens, P. asymbiotica, Xenorhabdus bovienii).

[APPLICATION OF PULSED FIELD GEL ELECTROPHORESIS FOR MOLECULAR TYPING OF CAUSATIVE AGENTS OF ESPECIALLY DANGEROUS INFECTIONS].

The macro-restriction analysis of the microorganism DNA with the use of gel electrophoresis in pulsed field (PFGE typing, pulse electrophoresis) is applied in molecular biology to study the clonal structure and typing of causative agents of infectious diseases. Determining the degree of the relationship and definition of epidemiological interrelations of studied isolates, as well as studying the evolutionary history of pathogens, is performed by comparing DNA restriction patterns. This review presents an analysis of the use of the pulse electrophoresis in molecular-epidemiological research and the study of phylogeny of especially dangerous infections, cholera, and plague. The possibility of genetic heterogeneity of the Vibrio cholerae and Yersinia pestis populations is demonstrated; territorial and epidemiological characteristics of the spread of different isolate pulso-types, problems and prospects of the PFGE typing method in the system of epidemiological surveillance of cholera and plague in Russia are discussed.

Chitin and Products of Its Hydrolysis in Vibrio cholerae Ecology.

The role of chitin and its hydrolysis products generated by Vibrio cholerae chitinases in mechanisms of its adaptation in water environments, metabolism, preservation, acquisition of pathogenic potential, and its epidemiological value are reviewed. Chitin utilization by V. cholerae as a source of energy, carbon, and nitrogen is described. Chitin association promotes biofilm formation on natural chitinous surfaces, increasing V. cholerae resistance to adverse factors in ecological niches: the human body and water environments with its inhabitants. Hydrolytic enzymes regulated by the corresponding genes result in complete chitin biodegradation by a chitinolytic catabolic cascade. Consequences of V. cholerae cell and chitin interaction at different hierarchical levels include metabolic and physiological cell reactions such as chemotaxis, cell division, biofilm formation, induction of genetic competence, and commensalic and symbiotic mutual relations with higher organisms, nutrient cycle, pathogenicity for humans, and water organisms that is an example of successful interrelation of bacteria and substratum in the ecology of the microorganism.

[Maldi-tof ms analysis for yersinia pestis, vibrio cholera, and francisella tularensis identification].

Numerous studies showed that a new technology for the clinical microbiology laboratories, Matrix-Assisted Laser Desorption Ionization--Time of Flight Mass Spectrometry (MALDI-ToF MS), allows fast, accurate, and effective identification of most clinically relevant microorganisms to be implemented. In the present review, we discuss applications of this approach for identification and typing of extremely dangerous pathogens--Yersinia pestis, Vibrio cholera, and Francisella tularensis, including the advantages and disadvantages of the method, sample preparation and biosafety problems.

[Multilocus sequence-typing of vibrio cholerae strains with various epidemic importance].

The allele polymorphism of the housekeeping genes (dnaE, lap, recA, pgm, gyrB, cat, chi, gmd) from the Vibrio cholerae strains with different epidemic importance (n = 41) isolated in Siberia and at the Far East during the cholera pandemic VII was tested. All toxigenic strains isolated at the period of epidemic complications irrespective of time and source of isolation were characterized by the identical allele profile and belonged to the same sequence-type. Nine sequence types were detected in non-epidemic isolates. The dendrogram clustering was associated with the serogroup and in some cases with the territory and time of isolation. The structure heterogeneity of the non-toxigenic V. cholerae housekeeping genes was in most cases caused by the synonymous nucleotide replacements (Dn/Ds < 1) indicating the prevalence of the negative V. cholerae at the analyzed genome sites. The revealed distinctions in the structure of housekeeping genes of the V. cholerae with different epidemic importance can be regarded as evidence of various evolutional directions in these strain groups.