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This study aimed to determine phenotypic and genotypic drug resistance patterns of Mycobacterium tuberculosis strains from children with tuberculosis (TB) in China and Russia, two high-burden countries for multi/extensively-drug resistant (MDR/XDR) TB. Whole-genome sequencing data of M. tuberculosis isolates from China (n = 137) and Russia (n = 60) were analyzed for phylogenetic markers and drug-resistance mutations, followed by comparison with phenotypic susceptibility data. The Beijing genotype was detected in 126 Chinese and 50 Russian isolates. The Euro-American lineage was detected in 10 Russian and 11 Chinese isolates. In the Russian collection, the Beijing genotype and Beijing B0/W148-cluster were dominated by MDR strains (68% and 94%, respectively). Ninety percent of B0/W148 strains were phenotypically pre-XDR. In the Chinese collection, neither of the Beijing sublineages was associated with MDR/pre-XDR status. MDR was mostly caused by low fitness cost mutations (rpoB S450L, katG S315T, rpsL K43R). Chinese rifampicin-resistant strains demonstrated a higher diversity of resistance mutations than Russian isolates (p = 0.003). The rifampicin and isoniazid resistance compensatory mutations were detected in some MDR strains, but they were not widespread. The molecular mechanisms of M. tuberculosis adaptation to anti-TB treatment are not unique to the pediatric strains, but they reflect the general situation with TB in Russia and China.

Ancient sublineage of the Mycobacterium tuberculosis Beijing genotype is endemic and prevalent in East Asia and rare in other world regions. While these strains are mainly drug susceptible, we recently identified a novel clonal group Beijing 1071-32 within this sublineage emerging in Siberia, Russia and present in other Russian regions. This cluster included only multi/extensive drug resistant (MDR/XDR) isolates. Based on the phylogenetic analysis of the available WGS data, we identified three synonymous SNPs in the genes Rv0144, Rv0373c, and Rv0334 that were specific for the Beijing 1071-32-cluster and developed a real-time PCR assay for their detection. Analysis of the 2375 genetically diverse M. tuberculosis isolates collected between 1996 and 2020 in different locations (European and Asian parts of Russia, former Soviet Union countries, Albania, Greece, China, Vietnam, Japan and Brazil), confirmed 100% specificity and sensitivity of this real-time PCR assay. Moreover, the epidemiological importance of this strain and the newly developed screening assay is further stressed by the fact that all identified Beijing 1071-32 isolates were found to exhibit MDR genotypic profiles with concomitant resistance to additional first-line drugs due to a characteristic signature of six mutations in rpoB450, rpoC485, katG315, katG335, rpsL43 and embB497. In conclusion, this study provides a set of three concordant SNPs for the detection and screening of Beijing 1071-32 isolates along with a validated real-time PCR assay easily deployable across multiple settings for the epidemiological tracking of this significant MDR cluster.

We performed synthesis of new nitrofuranyl amides and investigated their anti-TB activity and primary genetic response of mycobacteria through whole-genome sequencing (WGS) of spontaneous resistant mutants. The in vitro activity was assessed on reference strain

espanolEste articulo aborda la contribucion de MeteoGalicia en el campo de la educacion ambiental, a traves de su participacion en el Plan del Proyecto, con el programa MeteoEscolas. La metodologia seguida en MeteoEscolas es el aprendizaje basado en proyectos basado en el estudio interdisciplinario, en este caso de meteorologia y observacion climatologica, que involucra las diferentes materias que conforman el curriculo academico en las diferentes etapas educativas. El resultado de este trabajo conjunto sera un proyecto compuesto por actividades desarrolladas en el centro, con la participacion de toda la comunidad educativa. EnglishThis paper treats the work of MeteoGalicia, dependent of the department of environment, territory and housing, within the scope of the environmental education, through its participation in the plan Proxecta of the department of education, with MeteoEscolas. The methodology that is followed in MeteoEscolas is the projects based learning, which is based on the interdisciplinary studio, in this case of the meteorology and climatology, involving the different subjects that build up the academic curriculum in the educational stages. The result will be a project with the activities developed by the school center, with the participation of the whole educational community. galegoNeste articulo tratase da aportacion de MeteoGalicia no ambito da educacion ambiental, a traves da sua participacion no Plan Proxecta, co programa MeteoEscolas. A metodoloxia que se sigue en MeteoEscolas e a aprendizaxe baseada en proxectos tendo como eixo o estudio interdisciplinar, neste caso da meteoroloxia e observacion climatoloxica, implicando as distintas mateiras que forman o currriculum academico nas distintas etapas educativas. O resultado deste traballo comun sera un proxecto composto polas actividades desenvolvidas no centro, coa participacion de toda a comunidade educativa. portuguesEste artigo trata da contribuicao do MeteoGalicia no campo da educacao ambiental, atraves de sua participacao no Plano do Projeto, com o programa MeteoEscolas. A metodologia seguida no MeteoEscolas e a aprendizagem baseada em projetos, baseada em estudos interdisciplinares, neste caso de meteorologia e observacao climatologica, envolvendo as diferentes disciplinas que compoem o curriculo academico nas diferentes etapas educacionais. O resultado desse trabalho conjunto sera um projeto composto por atividades desenvolvidas no centro, com a participacao de toda a comunidade educacional.

The synthesis of 20 arylidenecamphors and 15 pyrimidines with camphane skeleton is described in the current report. A modified method for preparation of sterically hindered 2- aminopyrimidines in two steps was demonstrated. The evaluation of their in vitro activity against Mycobacterium tuberculosis H37Rv showed different MIC values (up to 0.91 μM for ketone 39). Compound 35 demonstrated moderate (8.23 μM), but sustainable activity toward a collection of drug-resistant M. tuberculosis strains. Many of the compounds (especially among 2-aminopyridines 42–56) exhibited good to excellent activity against different strains of pathogenic bacteria and fungi (MIC up to 0.60 μM for compound 50), compared with reference antibiotics. Many of the newly designed compounds possess also in vitro cytotoxicity., This study was supported by: Bulgarian National Science Fund- project KP-06-H39/7 and Spanish Ministry of Science, Innovation and Universities- Grant RTI2018-094629-BI00. MEDINA’s authors disclosed the receipt of financial support from Fundación MEDINA, a public-private partnership of Merck Sharp and Dohme de EspañaS.A./Universidad de Granada/Junta de Andalucía.

Mycobacterium tuberculosis is a highly studied pathogen due to public health importance. Despite this, problems like early drug resistance, diagnostics and treatment success prediction are still not fully resolved. Here, we analyze the incidence of point mutations widely used for drug resistance detection in laboratory practice and conduct comparative analysis of whole-genome sequence (WGS) for clinical M. tuberculosis strains collected from patients with pulmonary tuberculosis (PTB) and extra-pulmonary tuberculosis (XPTB) localization. A total of 72 pulmonary and 73 extrapulmonary microbiologically characterized M. tuberculosis isolates were collected from patients from 2007 to 2014 in Russia. Genomic DNA was used for WGS and obtained data allowed identifying major mutations known to be associated with drug resistance to first-line and second-line antituberculous drugs. In some cases previously described mutations were not identified. Using genome-based phylogenetic analysis we identified M. tuberculosis substrains associated with distinctions in the occurrence in PTB vs. XPTB cases. Phylogenetic analyses did reveal M. tuberculosis genetic substrains associated with TB localization. XPTB was associated with Beijing sublineages Central Asia (Beijing CAO), Central Asia Clade A (Beijing A) and 4.8 groups, while PTB localization was associated with group LAM (4.3). Further, the XPTB strain in some cases showed elevated drug resistance patterns relative to PTB isolates. HIV was significantly associated with the development of XPTB in the Beijing B0/W148 group and among unclustered Beijing isolates.

In this paper, the specificity of patent images was studied, a method was developed that uses a neural network for image preprocessing (classification) and their subsequent comparison, the architectures of neural networks for working with images and deep learning libraries were analyzed, and a comparative analysis of existing methods for searching and classifying patent images was carried out. A number of practical tasks have been completed for the search and collection of patent images, the selection of the main classes of patent images, the formation of a training sample; the training of the neural network for the recognition of the selected classes of patent images was carried out, the analysis of the trained model was carried out; a software module was created based on the developed method.

Mycobacterium tuberculosis strains of the early ancient sublineage of the Beijing genotype are mostly drug susceptible and mainly circulate in East Asia. We have recently discovered two clusters of this sublineage emerging in the Asian part of Russia (VNTR-defined 1071-32 and 14717-15 types) and, to our surprise, both were strongly MDR/XDR-associated. Here, we evaluated their pathogenic features. The clinical isolates and reference laboratory strain H37Rv were investigated in the C57BL/6 mouse model to assess their virulence and lethality properties. The BACTEC MGIT 960 system was used to study the in vitro growth characteristics. In the murine model, strains 396 (14717-15-cluster, from Buryatia, Far East) and 6691 (1071-32-cluster, from Omsk, Siberia) demonstrated contrasting properties. The 396-infected group had significantly higher mortality, more weight loss, higher bacterial burden, and more severe lung pathology. Furthermore, compared to the previously published data on other Russian epidemic Beijing strains (B0/W148, CAO, Central Asian Russian), strain 396 demonstrated the highest mortality. Under the in vitro growth experiment, cluster 14717-15 isolates had significantly shorter lag-phase. To conclude, low-virulent MDR strain 6691 belongs to the Beijing 1071-32-cluster widespread across FSU countries but at low prevalence. This corresponds to common expectation that multiple drug resistance mutations reduce fitness and virulence. In contrast, highly lethal and hypervirulent MDR strain 396 represents an intriguing Beijing 14717-15 cluster predominant only in Buryatia, Far East (16%), sporadically found beyond it, but not forming clusters of transmission. Further in-depth study of this most virulent Russian Beijing cluster is warranted.

The Mycobacterium tuberculosis Mlva cluster 1071-32 belongs to the early ancient sublineage of the Beijing genotype. We have recently demonstrated its association with pre-XDR/XDR, increase in prevalence in parts of Russia (Northwest, Center, Siberia) and sporadic isolation in eastern Europe (Lithuania, Albania). Here, we analyzed whole genome sequencing (WGS) data in order to gain insight into genomic background of the pathogenetic properties and dissemination pattern of this cluster. The initial WGS collection (this study, GenBank, Broad Institute, GMTV) included 3089 samples, in particular, 521 from East/Southeast Asia and 1067 from Russia. Of these, 117 genomes were assigned to the Beijing early ancient sublineage. The WGS-based phylogenetic analysis subdivided samples into several clades. A large stabnd-alone branch included Russian strains of the VNTR cluster 1071-32; all were pre-XDR or XDR. Cluster-specific SNPs were determined: 15 in intergenic regions, 10 in PE_PGRS and PPE genes, 96 in other genes. Mutations leading to significant amino acid changes were found in genes of the following categories: intermediate metabolism and respiration (n=6), cell wall and cell processes (n=4), conserved hypotheticals (n=2), lipid metabolism (n=1). Three SNPs were selected for development of the real-time PCR method of detection of this resistant cluster. To conclude, the emerging highly drug-resistant Russian cluster of the M. tuberculosis Beijing genotype is characterized by a combination of potentially significant SNPs in the genes coding for intermediate metabolism and respiration and cell and cell wall processes. Supported by Russian Science Foundation (grant 19-14-00013).

The local situation with tuberculosis (TB) is shaped by the complex interplay of multiple factors related to both human host and Mycobacterium tuberculosis. We hypothesized that TB epidemiology in the rural regions in the Soviet Union was impacted by construction of the Gulag camps and significant incoming migration. This molecular M. tuberculosis study was conducted in 2017 in the Komi Republic in northern Russia, a region with high rate (26%) of primary multidrug-resistant (MDR) TB. MDR was detected in 30.8% (40/130) isolates; eight were extensively drug resistant. The Beijing genotype was predominant (56.2%). The main Beijing subtypes B0/W148 and 94-32 differed in the MDR rate, 83.3% and 27.2%, respectively. The non-Beijing isolates represented five genotypes (LAM, Ural, Haarlem, X, T). The proportion of Beijing B0/W148 in the "camp" cities (originated from Gulag camps) was twice as large as in other districts of the Komi Republic. To conclude, сirculation of the MDR-associated Beijing B0/W148 cluster critically influences the current situation with MDR-TB in this Russian region. The increased prevalence of B0/W148 in the urban setting on the whole, and in the "camp cities", in particular, indirectly points to the increased transmission capacity of this successful Russian strain of M. tuberculosis.

Whole-genome analysis of Mycobacterium tuberculosis isolates collected in Russia (N = 71) from patients with tuberculous spondylitis supports a detailed characterization of pathogen strain distributions and drug resistance phenotype, plus distinguished occurrence and association of known resistance mutations. We identify known and novel genome determinants related to bacterial virulence, pathogenicity, and drug resistance.

The activity of the design department (DD) is currently one of the most significant areas in the field of nuclear energy, the defense and rocket, and space industries. This area characterized by large volumes of used economic, technological, industrial and other resources. One of the tasks of the DD is to evaluate information assets (IA) and protect them from threats. The objectives of this article are to develop a method for assessing DD’s IA, as well as discussing the possibility of using this method in practice and the risks to which the DD is exposed unless special measures are taken to evaluate IA and threat analysis. Studies related to the assessment of the importance of information IA, the development of a threat model, and damage prediction investigated. The specificity of DD and processing of IA has been determined. In this paper, it is proposed to use the set-theoretic model for evaluating the IA of the DD that takes into account their structure, attributes, and life cycle. To assess the damage caused by the threat, it proposed to use the method of ALE. For the first time, a method for evaluating the IA of DD proposed. This method implemented in software. The software implementation successfully tested in an enterprise that includes the DD. The investigated problem studied quite recently. Therefore, there are open questions such as the classification of information assets of the DD and their categorization (confidentiality stamping), analysis of the life cycle of IA and threats, determination of the criticality level of the DD’s IA, assessment of damage from the realization of the threat. The prospect for the development of the research is the development of a threat model for information systems of the DD.

In a series of recently conducted forensic studies related to methods of a criminal investigation in the sphere of computer information, scientifically interesting data about the constancy of connections of particular characteristics of these criminal actions have been obtained. On their basis, a researcher is supposed to build typical and then individual models of an event being investigated. As practice shows, such a problem is quite complicated for officials of preliminary investigation agencies since they have to choose necessary data from legal regulations and a wide range of reference materials (the Criminal Code, the Code of Criminal Procedure, federal laws, and other statutory acts regulating public relations in the sphere of information, information technologies, and protection of information, forensic scientific products, etc.), most of which don’t apply to a particular case under investigation. It appears that in present-day conditions of science and technology development this problem should be solved on the basis of forensic computer modeling.

Aim and objectives: Despite the availability of sufficiently effective treatment regimens, prevention and control, the widespread transmission and prevalence of resistant variants of Mycobacterium tuberculosis, which do not respond to any of the commercial drugs, remains a serious health problem for both developed and developing countries. Multidrug-resistant (MDR) strains present new threat to health security. This situation is of global concern due to increased human mobility, partly illegal and spontaneous. Patients infected with drug resistant strains are hard or impossible to be cured, and their treatment is more toxic and expensive. The urgent need of new antimycobacterial agents and development pathways is becoming more and more apparent. Methods: In this study, we synthesized new compounds containing 2-aminobutanol and camphane moieties and evaluated them with the reference strain and clinical M. tuberculosis isolates from a high-burden country, Russian Federation. All four compounds were obtained by simple synthetic procedures as pure diastereoisomeres and characterized by mass spectrometry, nuclear magnetic resonance, melting points and elemental analysis. The cell lines HEP-G2, HaCaT, HEK293 and CCL-1 were used to study the cytotoxicity and metabolism of tested compounds. The MTT colorimetric test was used to determine the average inhibitory concentrations of 50% (IC50). The CFU test was used to assess the long-term antiproliferative effect of the compounds. In vitro antimycobacterial activity of all compounds was tested by resazurin microtiter assay (REMA). All compounds were tested against reference strain H37Rv and 11 clinical isolates. Results: The isolates represented different clusters within clinically/epidemiologically important Beijing genotype (B0/W148, 94-32, Central Asian Outbreak, ancestral sublineage) and differed in phenotypic drug resistance (one extensively drug resistant, six multidrug resistant, two polyresistant, and two susceptible to all tested drugs). The most efficient compound showed a wide range of MIC that varied from 3.1 μg/ml (H37Rv, ancestral Beijing, most of B0/W148 isolates) to 100 μg/ml (other isolates). Conclusions: To conclude, this study highlighted a key importance of testing new anti-TB compounds not only with reference laboratory strain H37Rv that belongs to the phylogenetically marginal branch of M. tuberculosis, but on the clinical drug-resistant isolates circulating in the countries with high burden of MDR-TB. Acknowledgements. This study was supported by the Russian Science Foundation (grant 19-15-00028). Partial support was received from the Bulgarian National Science Fund (grant KP-06-H39-7).

Aim and objectives: Emergence of multi-drug resistant (MDR) and extensively drug-resistant (XDR) Mycobacterium tuberculosis strains requires use of novel compounds. New drugs require long-term treatment regimen and, consequently, long-term drug selective pressure gives more time to mycobacteria to develop resistance. At the same time, new compounds are expensive and, albeit efficient, they are not routinely used in the Russian Federation which situation with tuberculosis is characterized by high and increasing prevalence of multidrug resistant strains. Bedaquiline is a novel and promising anti-TB drug. Substitutions in the M. tuberculosis atpE gene prevent interaction of the drug with its target, ATP synthase and were associated with bedaquiline resistance. Mutations in two other genes involved in intermediary metabolism and respiration were also linked to the bedaquiline resistance: pepQ coding for probable cytoplasmic peptidase and atpC coding for probable ATP synthase epsilon chain. Mutations in the repressor gene mmpR (Rv0678) lead to increased expression of the efflux genes mmpL5 and mmpS5, providing export of bedaquiline. Finally, mutations in Rv1979c coding for possible conserved permease involved in transport of amino acids across the membrane were also shown in bedaquiline-resistant isolates. Perchlozone is a new thiosemicarbazone that is similar to thiacetazone and approved for treatment of MDR-TB in Russia. It is a prodrug activated by monooxygenase EthA and its target is FASII dehydratase complex HadABC. A cross-resistance to TAC and PCZ was shown by in vitro experiments and was mediated by both ethA and hadA mutations. On the other hand, EthA is known to activate second-line drugs ethionamide (ETH) and prothionamide (PTH) and ethA or ethR mutations were described as ETH/PTH resistance mechanism. Linezolid is other important anti-TB drug used for treatment of MDR/XDR-TB. The drug binds to the ribosome at the peptidyl-transferase center, and in vitro and clinical studies described mutations in the 23S rRNA and L3 ribosomal protein genes in resistant strains. Russia is a recognized hot spot for MDR-TB. TB incidence is decreased in the European Russia and is very high in Siberia and Far East, however, MDR TB is high and increasing in all parts of the country. For example, in Northwest Russia, TB incidence is 31.3/100,000 and MDR rate in primary TB - 27.4%. Kaliningrad is the westernmost Russian region and has a unique geographic position, separated from the mainland Russia by Lithuania, Poland and the Baltic Sea. The region is characterized by high TB incidence (38.8/100,000) and very high rate of MDR among newly diagnosed TB patients (30.5%). The M. tuberculosis population structure is dominated by the Beijing genotype. In this study, we aimed to gain insight into molecular basis of M. tuberculosis resistance to bedaquiline, perchlozone and linezolid in the Kaliningrad region in Russia. We used whole-genome sequencing of multiple consecutive isolates from patients spanning long time period in order to gain comprehensive and quantitative view on the real-time mycobacterial adaptation to the human host. This study included patients admitted to the clinical tuberculosis dispensary in Kaliningrad whose treatment regimen included second-line, repurposed and new anti-TB drugs, including BDQ, PCZ and LZD. All samples used in this study were coded and lacked personal information about patients. All personal data were treated anonymously and, according to the regulations of the St. Petersburg Research Institute of Phthisiopulmonology, no additional ethical approval was required. Methods: M. tuberculosis isolates were recovered from sputum at the 1-3 months intervals. M. tuberculosis drug susceptibility testing for first- and second-line drugs was performed using modified proportion method on Middlebrook 7H9 liquid culture and Bactec MGIT 960 system (Becton Dickinson, Sparks, Md.) according to the manufacturer's instructions. The laboratory is externally quality assured by the System for External Quality Assessment “Center for External Quality Control of Clinical Laboratory Research” (Moscow, Russia). Whole genome sequencing was performed at the MiSeq platform (Illumina). The fastq files were aligned to the complete genome of the reference strain H37Rv (NC_00962.3) using Geneious 9 program (Biomatters, New Zealand) and additionally checked with PhyReSE online tool ( https://bioinf.fz-borstel.de/mchips/phyresse/ ). Results: Mutations in Rv0678 (mmpR) were most frequent in BDQ resistant strains and different types of mutations were found (non-synonymous substitutions, frameshift, multi-codon indels). In some cases, different mutations coexisted and heteroresistance was observed in all cases, i.e., simultaneous presence of wild type and mutants alleles (which correlates with previous studies). Only in one patient, atpE mutation emerged in M. tuberculosis isolate during treatment. Analysis of genes associated with PCZ resistance (ethA, ethR, and hadABC) identified three mutations in ethA and one mutation in hadA. The ethA frameshift mutation (genome position 4327363 CT>C) was detected in isolates from patients who did not receive previous PCZ treatment but were all phenotypically resistant to PTH or ETH. Thus we assumed that this mutation emerged as PTH/ETH resistance mutation. No mutations in rrl, rplC, rplD genes associated with LZD resistance were found. Conclusions: To conclude, mutations in Rv0678 present a main mechanism of bedaquiline resistance in the Russian setting; development of resistance during treatment is not rare as bedaquiline resistance mutations emerged in 40% of patients. Furthermore, development of BDQ resistance 2-3 months after stop of treatment is possible, due to long half-life of the drug. With regard to PCZ resistance, presence of cross-resistance mutations to both PCZ and ETH/PTH suggests that these mutations should be assessed before inclusion of PCZ in the treatment regimen. A large prospective study of the more diverse M. tuberculosis collection is warranted to elucidate the development of resistance to

The goal of this work was to obtain genotypic characteristics of multidrug-resistant (MDR) strains of Mycobacterium tuberculosis (i.e., strains that are resistant at least to rifampicin and isoniazid) isolated from consumptives in Northwestern Russia in 2011–2012. Spoligotyping of 195 strains of Mycobacterium tuberculosis revealed 14 spolingotypes belonging to genetic families Beijing (n = 162), LAM (n = 15), H3/URAL (n = 14), T, Harleem, and X. Spolingotypes SIT1 (Beijing), SIT42 (LAM), and SIT62 (H3/URAL) were predominant. Regardless of genotype, all studied strains were resistant to streptomycin. Multidrug-resistant strains were resistant to ethionamide (56%), amikacin (31%), kanamycin (40%), and capreomycin (33%). The fractions of the strains resistant to ethambutol were 71 (n = 115) and 42% (n = 14) among the Beijing and nonBeijing strains, respectively (p < 0.05). Representatives of the Beijing genetic family remain predominant in Northwestern Russia in the population of multidrug-resistant strains of Mycobacterium tuberculosis (83%).

Mycobacterium tuberculosisis a highly studied pathogen due to public health importance. Despite progress in M.tuberculosis genome diversity analysis, there remain insufficient data on genome analysis of M.tuberculosis strains associated with pulmonary vs. extrapulmonary TB (PTB or XPTB respectively) tissue localization. Here we conduct comparative analysis of whole-genome sequence (WGS) for clinical M.tuberculosis strains collected from patients with PTB (n=72) and XPTB (n=73) localization. We further analyze the incidence of point mutations widely used for drug resistance detection in laboratory practice. M.tuberculosis isolates were collected from patients with varying clinical status from 2007 to 2014 in Russia. Bacterial DNA was extracted and sequenced using MiSeq platform (Illumina). WGSdata allowed identifying M.tuberculosis substrains associated with distinctions in the occurrence in PTB v.s. XPTB cases. There occurred little statistically significant specific DNA variants or genotypes diagnostic of tissue distribution, however phylogenetic analyses did reveal M.tuberculosis genetic substrains associated with TB localization. XPTB was associated with Beijing CAO, A and 4.8 groups, while PTB localization was associated with group LAM (4.3). Further, XPTB strain in some cases showed elevated drug resistance patterns relative to PTB isolates. HIV is significantly associated with the development of XPTB in the Beijing B0/W148 group and among unclustered Beijing isolates. This research analysis pinpointed genomic markers identified in XPTB and PTB with detailed characterization of drug-resistance markers of Russian M.tuberculosis isolates. We suggest that further comprehensive analysis of bacterial and human biological signatures might allow for better understanding consistent pattern of XPTB development.

Extrapulmonary and, in particular, spinal tuberculosis (TB) constitutes a minor but significant part of the total TB incidence. In spite of this, almost no studies on the genetic diversity and drug resistance of Mycobacterium tuberculosis isolates from spinal TB patients have been published to date. Here, we report results of the first Russian and globally largest molecular study of M. tuberculosis isolates recovered from patients with tuberculous spondylitis (TBS). The majority of 107 isolates were assigned to the Beijing genotype ( n = 80); the other main families were T ( n = 11), Ural ( n = 7), and LAM ( n = 4). Multidrug resistance (MDR) was more frequently found among Beijing (90.5%) and, intriguingly, Ural (71.4%) isolates than other genotypes (5%; P < 0.001). The extremely drug-resistant (XDR) phenotype was exclusively found in the Beijing isolates ( n = 7). A notable prevalence of the rpoB531 and katG315 mutations in Beijing strains that were similarly high in both TBS (this study) and published pulmonary TB (PTB) samples from Russia shows that TBS and PTB Beijing strains follow the same paradigm of acquisition of rifampin (RIF) and isoniazid (INH) resistance. The 24-locus mycobacterial interspersed repetitive unit–variable-number tandem-repeat (MIRU-VNTR) subtyping of 80 Beijing isolates further discriminated them into 24 types (Hunter Gaston index [HGI] = 0.83); types 100-32 and 94-32 represented the largest groups. A genotype of Russian successful clone B0/W148 was identified in 30 of 80 Beijing isolates. In conclusion, this study highlighted a crucial impact of the Beijing genotype and the especially prominent role of its MDR-associated successful clone B0/W148 cluster in the development of spinal MDR-TB in Russian patients.

In this study, (Gd1−xYx)F3 (x=0.2, 0.4, 0.5 0.6 and 0.8) crystals were grown by the μ-PD method. Neither visible inclusions nor cracks were observed in (Gd1−xYx)F3 (x=0.4, 0.5 and 0.6) crystals. Lattice constants increase linearly as the Y3+ concentration decreases. Using (Gd1−xYx)F3 as a host material, the Sr0.5% Ce1%, 3%, 5% and 7% co-doped Gd0.5Y0.5F3 single crystal was prepared. In this crystal, Ce3+-perturbed luminescence at 380 nm is observed with 31.8 ns photoluminescence decay time. Under X-ray excitation rather high radioluminescence intensity of this band was obtained exceeding that of BGO scintillator. The energy transfer in the sequence of regular Ce3+→ (Gd3+)n→ perturbed Ce3+ sites was evidenced though observation of decay time shortening of the regular Ce3+ and Gd3+ centers and intensity change between Gd3+ and Ce3+-perturbed emission.

The goal of this work was to study the genotypic characteristics of the multidrug-resistant (MDR, i.e., resistant to at least rifampicine and isoniazid) Mycobacterium tuberculosis strains isolated in 2011-2012 from tuberculosis (TB) patients in the Northwest Russia. Spoligotyping of 195 M. tuberculosis isolates identified 14 different spoligotypes and assigned isolates to the genetic families Beijing (n = 162, 83%), LAM (n = 15), H3/URAL (n = 14), as well as T, Haarlem and X. Spoligotypes SIT1 (Beijing), SIT42 (LAM) and SIT262 (H3/URAL) were the most prevalent. Irrespective to the genotype, all the isolates were resistant to streptomycin. The multidrug resistance was accompanied by the resistance to ethionamide (56%), amikacin (31%), kanamycin (40%), and capreomycin (33%). The ethambutol resistance was found in 71% (n = 115) and 42% (n = 14) of the Beijing and non-Beijing strains, respectively (p < 0.05). In conclusion, the multidrug resistant M. tuberculosis population circulating in the Northwest Russia continues to be dominated by the Beijing family strains.

This study reviews the epidemiology of thyroid cancer during childhood from the environs of Gomel in Belarus and the clinical data of 64 children aged 4 to 16 years from this area who had been diagnosed with differentiated thyroid carcinoma following the nuclear accident of Chernobyl. One case of thyroid cancer in children (aged < 15 years at diagnosis) was observed during the period 1981-1985 (rate = 0.5; expressed as annual averages per million children under age 15 years in the region of Gomel and period identified) before the Chernobyl accident. Twenty-one cases of thyroid cancer in children were observed during 1986-1990 (rate = 10.5) and 143 (rate 97) during 1991-1994 after the Chernobyl accident. During the first 7 months of 1995, there were 33 more cases of thyroid cancer observed in children. Three children with thyroid cancer were born since 1986 in the Gomel region. A total of 64 children aged 4 to 16 years from this area who had been diagnosed with differentiated thyroid carcinoma had been reviewed by us during the period May to November 1994. The female/male ratio was 1.4:1.0. At the time of the first diagnosis the mean age of the children was 9.4 +/- 2.8 years, and at the time of the accident their mean age was 3.8 +/- 2.4 years. More than 90% of the patients were less than 6 years of age and 3 were still in utero at the time of the accident. The period of latency between the accident and the first diagnosis was 5.6 +/- 1.5 years. Their ages at the time of the first diagnosis and their ages at the time of the accident were significantly correlated (p = 0.001); there was no significant correlation between the age of each child at the time of the accident and the latent period before the onset of carcinoma. The aggressiveness of the tumor, evaluated on the basis of T stage, lymph node status, and lung metastases, did not correlate with age at the time of the first diagnosis or with the age at the time of the accident. The susceptibility of the thyroid to the carcinogenetic effects of radiation, particularly during the first years of life (< 5 years) has clearly been demonstrated. However, there appears to be no correlation between the aggressiveness of the tumor and the age of the patients.

Background Russian Republic of Karelia is located at the Russian-Finnish border. It contains most of the historical Karelia land inhabited with autochthonous Karels and more recently migrated Russians. Although tuberculosis (TB) incidence in Karelia is decreasing, it remains high (45.8/100 000 in 2014) with the rate of multi-drug resistance (MDR) among newly diagnosed TB patients reaching 46.5 %. The study aimed to genetically characterize Mycobacterium tuberculosis isolates obtained at different time points from TB patients from Karelia to gain insight into the phylogeographic specificity of the circulating genotypes and to assess trends in evolution of drug resistant subpopulations. Methods The sample included 150 M. tuberculosis isolates: 78 isolated in 2013–2014 (“new” collection) and 72 isolated in 2006 (“old” collection). Drug susceptibility testing was done by the method of absolute concentrations. Spoligotyping was used to test genotype-specific markers of a Latin-American-Mediterranean (LAM) family and its sublineages as well as a Beijing B0/W148-cluster. Results The largest spoligotypes were SIT1 (Beijing family, n = 42) and SIT40 (T family, n = 5). Beijing family was the largest (n = 43) followed by T (n = 11), Ural (n = 10) and LAM (n = 8). Successful Russian clone, Beijing В0/W148, was identified in 15 (34.9 %) of 43 Beijing isolates; all В0/W148 isolates were drug-resistant. Seven of 8 LAM isolates belonged to the RD115/LAM-RUS branch, 1 - to the LAM RD174/RD-Rio sublineage. MDR was found in Beijing (32/43), Ural (3/10), and LAM (3/8). In contrast, all T isolates were pansusceptible. Comparison of drug resistant subgroups of the new and old collections showed an increasing prevalence of the B0/W148 clonal cluster, from 18.0 % (mainly polyresistant) in 2006 to 32.6 % in 2014 (mainly MDR and pre-XDR). The West–east increasing gradient is observed for the Ural genotype that may be defined a ‘Russian’ strain. In contrast, the spoligotype SIT40 of the T family appears to be a historical Karelian strain. Conclusions Circulation of the MDR M. tuberculosis isolates of the Beijing genotype and its B0/W148 cluster continues to critically influence the current situation with the MDR-TB control in northwestern Russia including the Republic of Karelia. Revealed phylogeographic patterns of some genotypes reflect a complex demographic history of Karelia within the course of the 20th century.

Introduction The emergence and spread of multidrug-resistant and, more recently, extensively-drug resistant tuberculosis (MDR-TB and XDR-TB) is a major concern of public health that hampers efficient TB control. The Mycobacteriology Laboratory in the St. Petersburg Research Institute of Phthisiopulmonology serves as a reference center for northwest Russia (11 provinces; 1,700,000 sq. km; population of 13.5 million); additionally, the laboratory receives strains from other regions across Russia. This study aimed to perform a molecular characterization of XDR Mycobacterium tuberculosis isolates recovered from TB patients in northwestern Russia during the time period of 2011–2013. Materials and methods M. tuberculosis isolates from mainly pulmonary TB patients (extrapulmonary TB, n = 7) were identified and characterized using traditional biochemical methods, including susceptibility testing for 11 anti-TB drugs (streptomycin, isoniazid, rifampin, ofloxacin, kanamycin, amikacin, capreomycin, ethambutol, ethionamide, pyrazinamide, and PAS acid). Drug susceptibility testing (DST) was done using a method of absolute concentrations according to the guidelines of the Russian Ministry of Health (order No. 109 of 21 March 2003) and/or BACTEC MGIT 960 system according to the manufacturer’s recommendations. Extensive drug resistance was defined as recommended by WHO (World Health Organization) as resistance to isoniazid and rifampin (MDR) plus resistance to one of the fluoroquinolones and one of three injectable drugs (kanamycin, amikacin, capreomycin). The isolates were further subjected to spoligotyping followed by comparison with SITVIT_WEB and MIRU-VNTRplus databases. LAM family was defined by Rv0129c SNP analysis. Results A total of 115 XDR M. tuberculosis isolates were included in this study. They presented 41 different patterns of drug resistance. XDR was complemented with resistance to streptomycin (n = 114), and/or ethambutol (n = 93), ethionamide (n = 80), pyrazinamide (n = 43), or PAS acid (n = 40). Of the three injectable drugs, XDR was mainly due to kanamycin resistance alone (n = 39, 34%), followed by combined kanamycin and amikacin resistance (n = 13), amikacin and capreomycin resistance (n = 11), kanamycin and capreomycin resistance (n = 9). Spoligotyping assigned the isolates to 3 genetic families: Beijing (98; 87%), LAM (SIT42, n = 6; SIT252, n = 2; SIT496, n = 2) and Ural (SIT262, n = 7). SIT1 was predominant among Beijing isolates. The proportion of isolates resistant to any of 5 drugs (including, streptomycin, isoniazid, rifampin, ofloxacin and/or kanamycin, amikacin, and capreomycin) was 1.7% (n = 2), to 6 drugs – 7.8% (n = 9), to 7 drugs – 19.1% (n = 22), to 8 drugs – 26.9% (n = 31), to 9 drugs – 25.2% (n = 29), to 10 drugs – 12.1% (n = 14). Eight isolates of the Beijing genotype were resistant to all 11 tested drugs. Conclusions XDR M. tuberculosis population in northwestern Russia is heavily dominated by Beijing genotype isolates (87%). Acknowledgements Russian Science Foundation (Grant agreement 14-14-00292).