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.

IntroductionThe majority of the genetic variance of systemic lupus erythematosus (SLE) remains unexplained by the common disease-common variant hypothesis. Rare variants, which are not detectable by genome-wide association studies because of their low frequencies, are predicted to explain part of this ”missing heritability.” However, recent studies identifying rare variants within known disease-susceptibility loci have failed to show genetic associations because of their extremely low frequencies, leading to the questioning of the contribution of rare variants to disease susceptibility. A common (minor allele frequency = 17.4% in cases) nonsynonymous coding variant rs1143679 (R77H) in ITGAM (CD11b), which forms half of the heterodimeric integrin receptor, complement receptor 3 (CR3), is robustly associated with SLE and has been shown to impair CR3-mediated phagocytosis.MethodsWe resequenced ITGAM in 73 SLE cases and identified two previously unidentified, case-specific nonsynonymous variants, F941V and G1145S. Both variants were genotyped in 2,107 and 949 additional SLE cases, respectively, to estimate their frequencies in a disease population. An in vitro model was used to assess the impact of F941V and G1145S, together with two nonsynonymous ITGAM polymorphisms, A858V (rs1143683) and M441T (rs11861251), on CR3-mediated phagocytosis. A paired two-tailed t test was used to compare the phagocytic capabilities of each variant with that of wild-type CR3.ResultsBoth rare variants, F941V and G1145S, significantly impair CR3-mediated phagocytosis in an in vitro model (61% reduction, P = 0.006; 26% reduction, P = 0.0232). However, neither of the common variants, M441T and A858V, had an effect on phagocytosis. Neither rare variant was observed again in the genotyping of additional SLE cases, suggesting that there frequencies are extremely low.ConclusionsOur results add further evidence to the functional importance of ITGAM in SLE pathogenesis through impaired phagocytosis. Additionally, this study provides a new example of the identification of rare variants in common-allele-associated loci, which, because of their extremely low frequencies, are not statistically associated. However, the demonstration of their functional effects adds support to their contribution to disease risk, and questions the current notion of dismissing the contribution of very rare variants on purely statistical analyses.

The situation of small and medium-sized enterprises (SMEs) in the Russian Federation during the COVID-19 pandemic and the resulting restrictive measures introduced by the state are analyzed in the context of the long-term dynamics of the sector and changes in public policy towards SMEs over the last two decades. Based on the analysis of official statistics and business demographics in 2005–2020, it is revealed that unfavorable trend – first of all, a decline in business start ups and an increase in firms’ shutdowns – in the sector had developed already before the crisis of 2008 and further it continued to progress. The main factors for such negative dynamics have been analyzed by the authors both in Russia as a whole and in different groups of regions. The study shows that negative developments of the SME in the country should be understood in the context of negative socio-economic changes influenced by a series of established institutional traps which occurred during the systemic transition and the limited effectiveness of the measures applied to support SMEs. An analysis of the feasibility and efficiency of the urgent state support measures of small business under the CIVID-19 has been carried out. In conclusion, the authors draw practical recommendations on the principles of reloading the public policy toward SME which should be formed in future after the pandemic, as well as indicate possible junctions in their implementation.

Objective. To investigate genetic determinants of resistance to antimicrobial agents recommended for the treatment of syphilis and assess their prevalence in molecular subtypes of Treponema pallidum subsp. pallidum in the Russian Federation over the period of 2014-2017. Materials and Methods. A total of 161 clinical isolates of T. pallidum obtained from Tyva, Stavropol, Irkutsk, Kaluga, Novosibirsk and Omsk regions were included in this study. Genetic material of T. pallidum was detected by PCR with primers to polA gene. Determinants of resistance to penicillins (tromp1, tp47), tetracyclines (16S rRNA) and macrolides (23S rRNA) were determined using the gene sequence analysis. Molecular typing was performed by characterizing variable arp, tpr (E, G, J) and tp0548 genes according to the CDC protocol. Results of this study were compared to historical data on antimicrobial resistance of T. pallidum over the period of 2011-2012. Results. Analysis of tromp1 and tp47 gene sequences detected C22G and G208T substitutions, respectively. These polymorphisms were not significant for activity of the corresponding proteins, but differed the studied clinical isolates from the reference strain Nichols, therefore, linking them with epidemic genogroup T. pallidum Street Strain 14. Based on the analysis of G1058C mutation in the 16S rRNA gene, all clinical isolates obtained in 2014-2017 belonged to wild type, whereas this genetic determinant of resistance to tetracyclines was determined in 2 of 190 isolates obtained in 2011-2012. Also, A2059G/C mutation in the 23S rRNA gene was not found, whereas a significant A2058G substitution in this gene was determined in 4 isolates obtained in 2014-2017. Results of this study confirm sporadic resistance to macrolides in the Russian Federation, which was previously (2011-2012) found in 3 of 190 isolates of T. pallidum. A2058G mutation was detected predominantly in minor subtypes of T. pallidum (14 b/f, 14 b/g and 14 d/g) and was unrepresentative for molecular subtype 14 d/f which is a predominant one in the Russian Federation. Conclusions. The long-term use of penicillins for the treatment of syphilis did not result in emergence of T. pallidum resistance to this antibiotic class. An absence of genetic determinants of resistance to tetracyclines confirms them to be second-line drugs. A sporadic prevalence of determinants of resistance to macrolides requires they be used for the treatment of syphilis with caution.