Experimental Modeling of Leprosy in BALB/c, BALB/c Nude, CBA, and C57BL/6ТNF-/- Mice.

Leprosy was modeled in an experiment on BALB/c, BALB/cNude, CBA, and C57BL/6ТNF-/- mice using three Mycobacterium leprae strains obtained from patients with a diagnosis of A30 according to ICD-10 from different regions of the Russian Federation. Proliferation of M. leprae of the used strains showed a temporal-quantitative dependence on the used mouse line. CBA and BALB/cNude mice were optimal for strain R and BALB/c and BALB/cNude lines were optimal for strain I. BALB/cNude mice infected with strain I had low lifespan. M. leprae strain M showed low proliferation activity in BALB/cNude and C57BL/6ТNF-/- mice.

[The refinement of leprosy PCR diagnostics by the amplification of specie-specific repeated fragment of the Mycobacterium leprae genome.]

Certain level of new registered cases of leprosy in a number of endemic countries in the world, as well as growing rate of transboundary migratory flows, raise the issue of effective diagnosis of this disease in countries with sporadic incidence of leprosy, including the Russian Federation. The purpose of the study was to develop a highly sensitive PCR test for detecting the genetic material of Mycobacterium leprae and to compare the test robustness and sensitivity with the commercially available Leprosy Genesig Standard Kit (Primerdesign Ltd., UK). The proposed approach uses real time PCR of non-coding repeating element RLEP, unique for the M. leprae genome, using TaqMan probe. The high test specificity was shown using the reference DNA samples of pathogenic and conditionally pathogenic mycobacterium, as well and its comparison with single-copy genes of M. leprae (rrs, fbp, MntH) PCR detection. The use of a commercially available test system based on the single-copy rpoB gene detection provided 59.4% sensitivity to the detection of M. leprae in the clinical material, while the application of the developed approach increased this index to 96.8%. The developed PCR diagnostics test of leprosy is submitted for state clinical approval process, whereupon the practical use of the test diagnostics allows solving a wide range of tasks to identify and confirm new cases of leprosy, and monitoring both the effectiveness of leprosy treatment, and epidemiological (including transboundary) the spread of the disease.

Oligonucleotide Microchip for the Identification of Infectious Agents of Reproductive System with Simultaneous Analysis of Determinants of Resistance to Antimicrobial Substances.

We developed a multiplexed DNA microarray-based assay allowing identification of 12 causative agents of reproductive tract infections with the simultaneous detection of 47 genetic determinants of resistance to antimicrobial substances. The microarray was tested on 93 isolates of Neisseria gonorrhoeae, 32 isolates of Treponema pallidum and 29 samples of Ureaplasma spp./Mycoplasma spp. The N. gonorrhoeae isolates had multiple mutations in the penA, ponA, rpsJ, gyrA, parC, and mtrR genes; their prognostic value significantly increased when combinations of mutations were detected. In the analyzed T. pallidum isolates, single A2058G substitution in the 23S rRNA gene responsible for macrolide resistance was found. DNA sequences of Ureaplasma spp./Mycoplasma spp. were determined as wild type, which was not fully consistent with the results of analysis of their antimicrobial susceptibility.

MOLECULAR EPIDEMIOLOGY OF TREPONEMA PALLIDUM IN BORDER REGION OF RUSSIAN FEDERATION (TUVA REPUBLIC).

The 111 strains of T. pallidum subsp. pallidum collected in Tuva Republic in 2013-2016 were typed by the arp, tpr (E, G, J) and tp0548 genes. The 7 subtypes were identified, in which the 14 d/f type was predominant (90.1%). The minor subtypes 14 b/f, 14 c/f, 14 d/g and 14 i/f constituted 0.9-1.8%. Single strains of 4 d/f H 9 d/f types (each 0.9%) previously described in China were detected in 2015. Both 9 and 14 arp gene variants were found in 3 clinical specimens for the first time in 2015-2016. Similarities in the molecular epidemiology of syphilis in Tuva Republic and Russia were demonstrated as well as differences from the T. pallidum subsp. pallidum population in China and Western Europe.

[Identification of biological processes on the composition of the urine proteome in cosmonauts on the first day after long space flights].

There are several difficulties that impede medical support of space missions. We can assume that the long-term space missions will be accompanied by functional changes in the organism. These changes will be a natural reaction to the factors of space flight. Complex action of factors can lead to the development of both non-specific changes (general adaptation syndrome), and specific changes. We analyzed the physiological changes after long space flights and carried out the correlation of these changes with previously identified proteins in the urine. It is possible to determine the mechanisms of adaptation of the human organism to the conditions of life on Earth after a long stay in Earth orbit.