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EMERGENCE OF NEW INFECTIONS IN THE 21 CENTURY AND IDENTIFICATION OF PATHOGENS USING NEXT GENERATION SEQUENCING
стр.5-23
Makarov V.V., Khromov A.V., Guschin V.A., Tkachuk A.P.
Each new emerging infection may become a big challenge to the medical community. Changing environment, tropical deforestation, melting of the Antarctic ice, growing population density and uncontrolled use of antibiotics provoke emergence and evolution of pathogens. Epidemics caused by new strains of the influenza virus, respiratory syndromes associated with coronaviruses, outbreaks of hemolytic infections and antibiotic-resistant superbacteria are hazards to humans. Among high-priority measures for pathogen control that are yet to be taken is development of fast and accurate techniques for pathogen identification. Our review looks at the cases of new infections registered in the 21 century and explores feasibility of next generation sequencing for the detection and identification of new pathogens.
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GENETIC POLYMORPHISM OF STRAINS IN PATIENTS OF THE NEONATAL INTENSIVE CARE UNIT
стр.24-30
Gordeev A.B., Lyubasovskaya L.A., Rodchenko J.V., Dubodelov D.V., Mukosey I.S., Kochetkova T.O., Nikitina I.V., Ionov O.V., Zubkov V.V., Trofimov D.Y., Priputnevich T.V.
is a member of the normal bacterial flora of humans capable of causing potentially dangerous diseases in neonates with very or extremely low birth weight. The number of genes responsible for virulence and antibiotic resistance may vary in different strains. We sequenced isolates of to explore genetic diversity of 14 strains circulating in the Neonatal Intensive Care Unit of Kulakov Research Center for Obstetrics, Gynecology and Perinatology. Among the studied strains, 8 sequence types were identified, the most frequent being ST2 and ST59, both of which belong to the clonal complex СС2. Of 14 studied strains, 10 were of СС2 type. The studied strains revealed a variety of genes responsible for antibiotic resistance. We found 15 genes that provided resistance to aminoglycosides, beta4actam antibiotics, fusidic acid, macrolides, lincosamides, streptogramin B, tetracycline, and trimethoprim. We identified a number of genes associated with virulence (, , , ), whose frequency in the studied isolates was varied. The insertion element was detected in 9 strains, and 7 strains revealed the presence of the -operon responsible for the biosynthesis of the biofilm matrix proteins.
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POLYMORPHISM OF THE GENE IN THE CURRENTLY EXISTING STRAINS OF
стр.31-37
Chagina I.A., Perevarova Yu.S., Perevarov V.V., Chaplin A.V., Borisova O.Yu., Kafarskaia L.I., Afanas’ev S.S., Aleshkin V.A.
The pathogenic mechanism used by is attributed to the ability of the diphtheria toxin to disrupt protein synthesis in human cells. Diphtheria toxin production is regulated by the DtxR protein. The latter is involved in the iron-mediated repression of the toxin gene and coordinates activities of other genes essential for the survival of . The DtxR-encoding gene occurs in both toxigenic and non-toxigenic strains; therefore it can be used to analyze the population structure of the species. In our work we have studied 45 strains of isolated in the Russian Federation in 2010-2015. These strains were analyzed to reveal that gene is a highly conservative region of genome that can be found in all members of the studied species. The majority of the discovered polymorphisms were synonymous (16 of 18 single nucleotide polymorphisms identified). In spite of the low phylogenetic signal, the allelic variant of was associated with the strain’s phenotype (biovar, toxigenicity). The obtained data indicate the presence of aggressive negative selection aimed to maintain the existing protein sequence in the population. Based on the results, we recommend dtxR polymerase chain reaction as an additional technique for pathogen identification, which is especially relevant considering the increasing prevalence of the disease associated with non-toxigenic strains.
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AMINOPYRIDINE- AND AMINOPYRIMIDINE-BASED SERINE/THREONINE PROTEIN KINASE INHIBITORS ARE DRUG CANDIDATES FOR TREATING DRUG-RESISTANT TUBERCULOSIS
стр.38-43
Maslov D.A., Bekker O.B., Alekseeva M.G., Kniazeva L.M., Mavletova D.A., Afanasyev I.I., Vasilevich N.L., Danilenko V.N.
Tuberculosis (ТВ) is the world’s deadliest bacterial infection. Its causative agent evolves into rapidly spreading multidrug-resistant and extensively drug-resistant (MDR and XDR) strains, which complicates the treatment. Therefore, the use of novel target-specific chemical compounds is crucial for the development of effective antituberculosis agents. Serine/threonine protein kinases (STPKs) of are currently considered as attractive drug targets. In turn, aminopyridines and aminopyrimidines that have not been used for ТВ treatment so far exhibit inhibitory activity towards STPKs. In this study we screened 192 aminopyridine- and aminopyrimidine-based compounds using the test system designed to screen for active STPKs inhibitors. First, we selected 53 compounds with subinhibiting concentrations of up to 100 nmol/disk. Of them, 22 showed STPKs-inhibiting activity in the test system, which was confirmed in vitro on the PknA protein with a maximum of 26.9 ± 6.1 %. Toxicity testing was performed in vitro on human embryo fibroblasts using the MTT-assay. Ultimately, 3 relatively active and relatively non-toxic STPKs inhibitors were selected for further research as drug candidates for MDR-TB treatment.
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MIRU-VNTR GENOTYPING OF CLINICAL ISOLATES FROM MOSCOW REGION
стр.44-47
Shur K.V., Maslov D.A., Bekker O.B., Danilenko V.N.
Antibiotic selection pressure, genetic polymorphism as well as diversity of the immune status of the host and other selection factors continuously prompt , the tuberculosis causative agent, to evolve. Significant or insignificant mutations shape new (sub)lineages of the pathogen whose evolution can be understood only through analyzing and monitoring its genotypic diversity and properties of its lineages. In our study we used a set of 46 clinical isolates from Moscow region. The samples were typed using the standard 24-loci MIRU-VNTR technique. Beijing family isolates were shown to prevail in the collection (60.9 %), as well as Beijing-BOA/VI 48 subtype (60.7 % of total Beijing type samples); most of them (88,2 %) were multidrug-resistant resistant. The applied technique allowed us to detect one case of a mixed-strain infection.
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MANAGEMENT OF THE HIV-POSITIVE PREGNANT PATIENT WITH MARKED IMMUNODEFICIENCY AND MULTIPLE COMORBIDITIES
стр.48-54
Increasing HIV prevalence among women and growing numbers of HIV-positive patients who choose to become pregnant prompt a discussion of management strategies applied to such patients. In this work we analyze a case of a pregnant HIVpositive woman with marked immunodeficiency who started seeking medical advice only after she had developed severe life-threatening secondary conditions. We look at the progression of comorbidities that led to the death of the patient and her baby and evaluate the chosen treatment plan. We also propose recommendations for the management of patients with similar pathologies that include psychological care, vigilance against possible atypical progression of a comorbidity, such as tuberculosis, and extensive diagnostic evaluation.
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CLINICAL, IMMUNOLOGICAL AND VIROLOGICAL INDICATORS OF ANTIRETROVIRAL THERAPY EFFICIENCY
стр.55-61
Oleynik A.F., Fazylov V.H., Beshimov A.T.
Antiretroviral therapy (ART) for HIV-positive patients allowed labeling the disease a therapeutically controlled one. The main goal of ART is to prolong patient's life and preserve its quality. This is accomplished through viral load reduction (decrease of the number of HIV-RNA copies in blood plasma), which leads to the growing numbers of CD4-T-lymphocytes. However, ART can be ineffective. In 2010-1014, we conducted an observational cohort retro/prospective study aimed at learning how often ART can be ineffective from immunological (II), virological (VI) and clinical points of view. The study was carried out at the premises of the Republic Center of AIDS and Infectious Diseases (Kazan, Russia). The study included 341 adult HIV-positive patients subjected to ART at 3rd and 4th stages of disease's development, with the treatment virologically efficient at least during the first year. The observation period was 1 to 3 years. ART was considered II (immunologically inefficient) when the number of CD4 increased for less than 50 cells/mcl through the year with HIV completely suppressed. VI (virological inefficiency) of ART was registered if the number of HIV RNA copies was above the definition threshold after 6 months of treatment. ART was II in 14.0-15.9 % of cases after a year of treatment and in 22 % of cases after three years. It was noted that subsequent restoration of an adequate number of T-lymphocytes CD4 required they overcame the threshold of 100 cells/mcl within the 1st year of treatment. Virologically, ART was effective for 92.7 % for patients. Most (80 %) cases of VI of ART were results of patients' lax attitude towards treatment. Clinically, ART helped 91 % of patients; this result largely depended on the number participants for whom ART was II. II of ART is a risk factor, the risk being progression of the disease with active ART in the background and death of the HIV-positive individual. II of ART makes the risk of clinical progression of HIV 6.232 times higher (95 % Cl 3.106-12.51).
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DETECTING OCCULT HEPATITIS В WHEN TESTING DONATED BLOOD
стр.62-65
Eremeeva Zh.G., Fazylov V.H.
Individuals carrying occult (latent) hepatitis В pose epidemiological threat. Testing donated blood donors for surface antigen HBsAg (hepatitis В vims, HBV) only does not allow to assume the blood safe from the point of view of infections, which can result in post-transfusion transmission of infection. Lack of confidence here is due to the fact that the virus is present in the body even when HBsAg is negative. The study analyzes data of 61,155 blood donors of the Republican Blood Center (Kazan), collected in 2010-2014. The tests applied were those aimed at detecting HBsAg, anti-HBc-total, anti-HBc IgM (enzyme immunoassay), and determining DNA of the virus in the blood by polymerase chain reaction in "real time". It was found that donors with occult hepatitis В are identified each year, but their numbers decrease gradually. To prevent the spread of the virus it is recommended to add the anti-HBc-total test to the standard set of diagnostic tests.
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WOUND CARE WITH THE LEAF EXTRACT OF CECROPIN PI-PRODUCING TRANSGENIC KALANCHOE: HISTOLOGICAL FINDINGS
стр.66-73
Belous A.S., Shevelev A.B., Trubnikova E.V., Biryukova Yu.K., Mishina E.S., Loyko E.A., Lebedeva A.A., Zakharchenko N.S.
Management of purulent wounds is a problem that requires particular attention: wounds are a common injury type for which suppurative complications are frequent, mortality rates are high and antimicrobial therapy may be ineffective due to the presence of drug-resistant bacteria in the wound. In this work we have studied the effectiveness of wound treatment with the leaf extract of transgenic modified to produce antimicrobial peptide cecropin P1. Purulent wounds infected with were modeled in Wistar rats. Four groups of animals were formed, with 10 animals in each group. In all groups, the wounds were cleansed with 3 % hydrogen peroxide solution once a day; all groups except the controls received additional treatment. Group 2 received 10 % cefazolin solution, group 3 received kalanchoe juice, group 4 received the juice of cecropin P1-producing kalanchoe. Histologic stains of biopsy samples were performed after rats were sacrificed by anesthetic overdose on days 3, 10 and 14 after treatment onset. On day 3, wound dynamics was the same in all groups. On day 10 exudate was still observed in the controls; in group two exudation was almost finished and regeneration was about to begin; in groups 3 and 4 the wound defect was filled with granulation tissue. In spite of epidermal repair along the wound edges in groups 2 and 3, there still was some sloughing and granulation tissue was less mature than in group 4. We recommend conducting more extensive clinical research of the leaf extract of cecropin P1-containing transgenic .
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KEY PERFORMANCE INDICATORS FOR HEALTHCARE RESEARCH ORGANIZATIONS BETWEEN 2011 AND 2015
стр.74-78
Aniskevich A.S., Halfin R.A.
In this work we identify 16 key indicators to evaluate the performance of healthcare research organizations. These indicators comprehensively characterize such aspects of performance as research output and relevance, human resource development, integration into the international scientific community, distribution of scientific knowledge, promotion of the prestige of science, and resource provision. Below, we review the existing classification of medical research institutions and their key features. We present the results of the comprehensive performance evaluation of healthcare research organizations. We demonstrate the significance of the proposed indicators that accurately reflect the output and relevance of scientific research and stress that indicators currently used for performance evaluation are insufficient. We also emphasize the need for a systemic approach to personnel capacity assessment and confirm the importance of additional evaluation criteria that amount to 37.5 % of all key indicators.
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