The analysis of recently published data on hepatitis A virus (HAV) genome clinical features, molecular diagnostic value and cell culture propagation are reviewed. The growing need in the study of the genetic diversity of...The analysis of recently published data on hepatitis A virus (HAV) genome clinical features, molecular diagnostic value and cell culture propagation are reviewed. The growing need in the study of the genetic diversity of HAV isolates and the search of its possible new antigenic variants are underlined. The results of the cultivation of different HAV strains are analyzed for possible application in vaccine and diagnostic kit production.
Dentovskaia SV, Kopylov PKh, Ivanov SA
… +2 more, Ageev SA, Anisimov AP
Mol Gen Mikrobiol Virusol
· 2013 · PMID 24364139
Molecular mechanisms of the Yersinia pestis pathogenicity and peculiarities of maturation of specific immunity to plague are reviewed. The history and modern state of the plague vaccine development are described. Special...Molecular mechanisms of the Yersinia pestis pathogenicity and peculiarities of maturation of specific immunity to plague are reviewed. The history and modern state of the plague vaccine development are described. Special attention is focused on the prospects in the area of the plague vaccine development. The possible approaches to improvement of vaccine preparations are discussed.
Ozerova LV, Krasnikova MS, Troitsky AV
… +2 more, Solovyev AG, Morozov SY
Mol Gen Mikrobiol Virusol
· 2013 · PMID 24003511
The various classes of plant 21 - to 24-nt siRNAs derive from long dsRNA precursors that are processed by the ribonuclease Dicer-like (DCL). The species of ta-siRNA were originally discovered in Arabidopsis thaliana. Fou...The various classes of plant 21 - to 24-nt siRNAs derive from long dsRNA precursors that are processed by the ribonuclease Dicer-like (DCL). The species of ta-siRNA were originally discovered in Arabidopsis thaliana. Four gene families have been identified in Arabidopsis that each produces a number of ta-siRNAs: TAS1, TAS2, TAS3 and TAS4. The TAS3 genes encode tasiR-ARF species which target the mRNA of three Auxin Response Factor (ARF) genes (ARF2, ARF3/ETT and ARF4) for subsequent degradation. The function of TAS3 precursor RNA is controlled by two miR390 target sites flanking tandem of ta-siARF sequences. In this paper, we have studied the presence ofta-siARF RNA genes in the representatives of subtribe Senecioninae. Senecioneae is the largest tribe of Asteraceae, comprised of ca. 150 genera and 3,000 species which include many common succulents of greenhouses. Approximately one-third of species are placed in genus Senecio, making it one of the largest genera of flowering plants. However, there was no information on the structure of TAS genes in these plants. We revealed that the TAS3 species (TAS3-Sen1) in Senecio representatives was actively transcribed, and its homologues are distributed among many Asteracea plants and found to be similar to Arabidopsis AtTAS3a gene. We revealed several prematurely terminated transcripts of TAS3-Sen1. Finding the alternative shortened transcripts of TAS3-Sen1 lacking the 3'-terminal site cleaved by miR390 and retaining the 5'-terminal miR390 non-cleaved site suggested their using as decoys for the modulation of miR390 activity to regulate synthesis of ta-siARF RNAs in different Senecioninae species.
Dogen A, Kaplan E, Serin MS
… +6 more, Oksuz Z, Tezcan S, Aslan G, Sezgin O, Altintas E, Emekdas G
Mol Gen Mikrobiol Virusol
· 2013 · PMID 24003510
Hepatitis B virus (HBV) infection is a global health problem with more than 2 billion infected individuals. HBV infection leads to diverse outcomes ranging from acute to chronic hepatitis, which may result in severe comp...Hepatitis B virus (HBV) infection is a global health problem with more than 2 billion infected individuals. HBV infection leads to diverse outcomes ranging from acute to chronic hepatitis, which may result in severe complications as liver cirrhosis and hepatocellular carcinoma (HCC). HBV is one of the most important human DNA viruses having strong oncogenic potential. Recently, many studies have reported on HBV X gene and PreC promoter mutations associated with HCC. In order to detect the prevalence of HBx gene and PreC promoter mutations possibly related to HCC, we have analyzed sera samples collected from 61 patients with chronic hepatitis B. We have detected TI653 mutation in 1 of 61 (1,63%), A1896 mutation in 10 of 61 (16,39%), and T1762 - A1764 dual mutation in 4 of 61 (6,55%). T1653 and T1762- A1764 dual mutations were suggested significantly related to HCC in earlier reported studies. Our findings demonstrate that HBx gene and PreC promoter mutations related to HCC are present in our region and prospective clinical chord studies would be useful for better patient management and of early diagnosis of possible HCC cases.
Voronina AL, Chernukha MY, Shaginyan IA
… +9 more, Kunda MS, Avetisyan LR, Orlova AA, Lunin VG, Avakyan LV, Kapranov NI, Amelina EL, Chuchalin AG, Gintsburg A L
Mol Gen Mikrobiol Virusol
· 2013 · PMID 24003509
88 cultures of microorganisms referred to the Burkholderia cepacia complex (Bcc) during initial identification were analyzed by multilocus sequencing (Multilocus Sequence Typing, MLST). 13 genotypes (sequence type, ST) w...88 cultures of microorganisms referred to the Burkholderia cepacia complex (Bcc) during initial identification were analyzed by multilocus sequencing (Multilocus Sequence Typing, MLST). 13 genotypes (sequence type, ST) were detected, 9 of them (708, 709, 710, 711, 712, 714, 727, 728, 729) were identified for the first time. Two new alleles for the gene trpB (357, 358), one of the genes atpD (306) and gltB (352) were detected and registered. It was found that strains of 2 genotypes (711, 712) belong to the species B. multivorans, 1 (ST102) - B. contaminans, 1 (ST51) - B. stabilis, 1 (ST729) - B. vietnamiensis. Most strains of the sample, representing 8 genotypes (208, 241, 728, 727, 708, 709, 710, 714), belong to the species B. cenocepacia. Identified genotypes differ in the global spread of the world: 4 genotype (51, 102, 208, 241) have intercontinental distribution, 1 (712) - intra. It is shown that strains causing nosocomial infections, in most cases refer to genotypes 728 and 708. Epidemiologically significant in respect of patients with cystic fibrosis should recognize genotype 709, detected in strains isolated from patients in seven federal districts (FD) of Russia. The Bcc strains of genotypes 241 (B. cenocepacia) and 729 (B. vietnamiensis) were isolated from the patients of the Far Eastern FD. They are not typical for other FD Russia. The possibility of concomitant infection in cystic fibrosis patient with two genotypes 709 - epidemiologically significant and 708 - nosocomial, was indicated. The long-termpersistence of a single genotype strain in the organism of patients with cystic fibrosis was demonstrated as for Bcc species B. cenocepacia (ST 709), so for B. multivorans (ST712). The possibility of transferring the strain Bcc, typical for nosocomial environment to patient with cystic fibrosis at surgery was observed.
Rar VA, Epikhina TI, Pukhovskaya NM
… +2 more, Vysochina NP, Ivanov LI
Mol Gen Mikrobiol Virusol
· 2013 · PMID 24003508
Totally 484 Haemaphysalis japonica, 359 Haemaphysalis concinna and 221 Dermacentor silvarumn collected in Amur region and Khabarovsk Territory of the Russian Far East were examined on the presence of Anaplasmataceae bact...Totally 484 Haemaphysalis japonica, 359 Haemaphysalis concinna and 221 Dermacentor silvarumn collected in Amur region and Khabarovsk Territory of the Russian Far East were examined on the presence of Anaplasmataceae bacteria using nested PCR. All positive samples were characterized by analysis of the 16S rRNA gene and/or groESL operone nucleotide sequences. Forty nine H. japonica and three H. concinna were shown to contain DNA of two new Ehrlichia genetic variants. These genetic variants on the basis of the 16S rRNA gene and groESL operone nucleotide sequences analysis were most closely related to Ehrlichia spp. revealed in Haemaphysalis spp. ticks in Japan. Four H. concinna from Amur region were shown to contain DNA of a new Anaplasma bovis genetic variant, which corresponded to A. bovis genetic variant revealed in a red gray-backed vole and a Siberian chipmunk from the Far East. Three H. concinna and nine D. silvarum contained DNA of non-typical bacteria which can't be attributed to any Anaplasmataceae genera based on the determined sequences of the 16S rRNA gene fragments. The revealed non-typical bacteria on the basis of 16S rRNA gene sequences significantly differed from each other and didn't form a separate genetic group.
Malivanova TF, Skoromyslova EV, Yurchenko VA
… +3 more, Kononenko IV, Manzyuk LV, Mazurenko NN
Mol Gen Mikrobiol Virusol
· 2013 · PMID 24003507
TNF is an inflammatory cytokine that involved in pathogenesis of different malignancies. Promoter single nucleotide polymorphism -238(G/A)TNF (rs361525) is investigated for the detection of susceptibility to the infectio...TNF is an inflammatory cytokine that involved in pathogenesis of different malignancies. Promoter single nucleotide polymorphism -238(G/A)TNF (rs361525) is investigated for the detection of susceptibility to the infectious, autoimmune and oncological diseases. The goal of the study was to investigate the association of-238(G/A)TNF polymorphism (rs361525) with breast cancer (BC) prognosis. -238(G/A) TNF allelic variants were detected by PCR-RFLP. We failed to reveal the genotype distributions disparity among groups with different stages of the disease, ER, PR or Her2/neu positive versus negative status. The AG genotype frequency was about 10% and there were no BC patients with AA genotype in all separated groups. However the overall survival was significantly lower for AG then for GG carriers with II stage or ER-positive BC. Our data suggest that -238(G/A)TNF polymorphism perhaps is not involved in the initiation of malignancies but it is a substantial factor of BC prognosis.
Techniques for differentiating single bacterial isolates into intraspecies clusters corresponding to subspecies, biovars, and natural foci are reviewed. The techniques under consideration are reproducible under different...Techniques for differentiating single bacterial isolates into intraspecies clusters corresponding to subspecies, biovars, and natural foci are reviewed. The techniques under consideration are reproducible under different laboratory settings. A version of the intraspecies classification of Y. pestis that is in harmony with the International Code of Nomencláture of Bacteria is suggested.
Genetic predisposition of an individual patient should be taken in account to choose the proper treatment. Implementation to clinical practice requires the development of rapid, high-throughput, and easy assays intended...Genetic predisposition of an individual patient should be taken in account to choose the proper treatment. Implementation to clinical practice requires the development of rapid, high-throughput, and easy assays intended to detect single nucleotide polymorphisms. A detection kit intended to identify the hemostasis and folate cycle gene mutations G20210A FII, G1691A FV, G10976A FVII, G103T FXIII, C807T ITGA2, T1565C ITGB3, 5G(-675)4G PAI, G(-455)A FGB, C677T and A1298C MTHFR, A2756G MTR, A66G MTRR was suggested in this work. The method is based on the polymerase chain reaction and subsequent melt curve analysis of the complexes of amplicons with specific probe. Three single nucleotide polymorphisms can be identified in one tube using our detection kit that increases the productivity of the analysis in the clinical use. Different types of biological samples (buccal epithelium, saliva, plasma, serum, and urogenital swabs) can be used as the initial material for DNA isolation and further analysis by the method developed in this work.
The goal of this work was to determine the rate of the polymorphism of the genes-antagonists of receptor IL-1 (IL-1RA) and TNF-alpha in patients with gastritis and duodenal ulcer associated with Helicobacter pylori. The...The goal of this work was to determine the rate of the polymorphism of the genes-antagonists of receptor IL-1 (IL-1RA) and TNF-alpha in patients with gastritis and duodenal ulcer associated with Helicobacter pylori. The receptors were tested for the clinical manifestation of the disease. A total of 126 patients with different gastroduodenal pathology and H. pylori in autopsy were tested. The results of this work demonstrated a correlation between the risk of duodenal ulcer and allele A of gene TNF-alpha in position 308 in the patients. The analysis of the gene-IL-1RA polymorphism demonstrated statistically significant difference between the patients in the frequency of the genotype 2/l. The results of this work showed that parallel typing of the genes of H. pylori in virulence was required for characterization of the bacteria-patient association. The correlation between the results of the typing with polymorphism of genes of cytokines in patient autopsy was also required.
Kuleshevich EV, Savicheva AM, Arzhanova ON
… +1 more, Suvorov AN
Mol Gen Mikrobiol Virusol
· 2013 · PMID 23785787
Streptococcus agalactiae or group B streptococci (GBS) is the major cause of various diseases of the newborns and the adults. The genome of GBS contains from 14 to 15 mobile genetic elements (pathogenicity islands, PAI)....Streptococcus agalactiae or group B streptococci (GBS) is the major cause of various diseases of the newborns and the adults. The genome of GBS contains from 14 to 15 mobile genetic elements (pathogenicity islands, PAI). It is well-known that many GBS virulence determinants are localized not in the core genome, but on the pathogenicity islands. The goal of this work was to determine the distribution and genetic organization of PAI XII containing virulence genes sspB1, scpB, lmb among the clinical strains of GBS. 74 clinical strains of GBS were analyzed using PCR with primers corresponding to the genes of the virulence factors located on PAI XII. The pathogenicity island XII was determined only in 22% of the clinical strains. The genetic organization of the island was different between strains. There was no correlation between the presence of PAI and the serotype of GBS.
The comparative analysis of the associations between G20210A polymorphism of F2 gene, G1691A polymorphism of F5 gene, -5T/C polymorphism of gene GP1BA, I/D polymorphism of gene ACE and the risk of development of the stro...The comparative analysis of the associations between G20210A polymorphism of F2 gene, G1691A polymorphism of F5 gene, -5T/C polymorphism of gene GP1BA, I/D polymorphism of gene ACE and the risk of development of the stroke in two ethnical samplings--Russian and Ukrainian populations--was conducted. It was shown that the patients of the Russian population with genotype DD have a higher level of the risk of ischemic stroke development (OR = 1.4, 95% CI [1.05; 1.78], p = 0.02), whereas genotypes I/I and I/D are associated with the lower level of risk of ischemic stroke (OR = 0.7, 95% CI [0.56; 0.95], p = 0.02). In the Ukrainian ethnical sampling, differences in distribution of genotypes and alleles frequencies between patients with stroke and healthy persons upon given polymorphic locus are not significant, and I/D polymorphism of gene ACE is not associated with the risk of development of the stroke (OR = 0.8, 95% CI [0.48; 1.32], p = 0.45). The G20210A polymorphism of gene F2, G1691A polymorphism of gene F5, -5T/C polymorphism of gene GP1BA are not associated with the risk of stroke in two ethnical samplings.
The DNA polymerase iota (Pol iota), which has some peculiar features and is characterized by an extremely error-prone DNA synthesis, belongs to the group of enzymes preferentially activated by Mn2+ instead of Mg2+. In th...The DNA polymerase iota (Pol iota), which has some peculiar features and is characterized by an extremely error-prone DNA synthesis, belongs to the group of enzymes preferentially activated by Mn2+ instead of Mg2+. In this work, the effect of Mn2+ on DNA synthesis in cell extracts from a) normal human and murine tissues, b) human tumor (uveal melanoma), and c) cultured human tumor cell lines SKOV-3 and HL-60 was tested. Each group displayed characteristic features of Mn-dependent DNA synthesis. The changes in the Mn-dependent DNA synthesis caused by malignant transformation of normal tissues are described. It was also shown that the error-prone DNA synthesis catalyzed by Pol iota in extracts of all cell types was efficiently suppressed by an RNA aptamer (IKL5) against Pol iota obtained in our work earlier. The obtained results suggest that IKL5 might be used to suppress the enhanced activity of Pol iota in tumor cells.
Sergeeva EI, Ternovoĭ VA, Demina OK
… +6 more, Demina AV, Korneev AN, Shikov AN, Berillo SA, Agafonov AP, Sergeev AN
Mol Gen Mikrobiol Virusol
· 2013 · PMID 24645276
The real time PCR assay targeting influenza A and B virus, 5 subtypes of influenza A virus (seasonal H1N1, pandemic H1N1 (2009), seasonal H3N2, pathogenic for human subtypes of avian influenza H5 and H7), respiratory syn...The real time PCR assay targeting influenza A and B virus, 5 subtypes of influenza A virus (seasonal H1N1, pandemic H1N1 (2009), seasonal H3N2, pathogenic for human subtypes of avian influenza H5 and H7), respiratory syncytial virus, and adenovirus was developed. The analytical sensitivity of the developed assay was 1 x 10(3) genome equivalents per ml. The diagnostic sensitivity of the method was 1 x l0(3)-10(4) viral particles per ml. Experiments with human DNA/cDNA and viral cDNA showed a markedly high diagnostic specificity of the developed PCR assay. In the assay of the developed PCR test, 50 nasopharyngeal swab specimens were tested. The etiology was identified in 33 samples.
Several distinct methods currently used for the Crimean-Congo Hemorrhage Fever diagnosis (CCHF) were suggested in this work. We demonstrated that the ELISA-based diagnostic kits, which are based on CCHFV recombinant anti...Several distinct methods currently used for the Crimean-Congo Hemorrhage Fever diagnosis (CCHF) were suggested in this work. We demonstrated that the ELISA-based diagnostic kits, which are based on CCHFV recombinant antigens produced in E. coli cells, still possessed a few substantial shortcomings, which are yet to be addressed. In this work we presented the development of the unique CCHFV detection system fully based on reverse transcription--nested two-step polymerase chain reaction (RT-PCR). Our RT-PCR-based diagnostic kit for the CCHFV detection is now commercially available. We also developed a simple screening method for the samples, potentially containing CCHFV, which is based on restriction fragment length polymorphism (RFLP) amplicons analysis and allows for preliminary genotyping of the CCHFV isolates.
Medkova AIu, Siniashina LN, Rumiantseva IuP
… +3 more, Voronina OL, Kunda MS, Karataev GI
Mol Gen Mikrobiol Virusol
· 2013 · PMID 24645274
The duration of the persistence and dynamics of accumulation of insertion bvg- Bordetella pertussis mutants were studied in lungs of laboratory mice after intranasal and intravenous challenge by virulent bacteria of the...The duration of the persistence and dynamics of accumulation of insertion bvg- Bordetella pertussis mutants were studied in lungs of laboratory mice after intranasal and intravenous challenge by virulent bacteria of the causative agent of whooping cough. The capability of the virulent B. pertussis bacteria to long-term persistence in the body of mice was tested. Using the real-time PCR approximately hundred genome equivalents of the B. pertussis DNA were detected in lungs of mice in two months after infection regardless of the way of challenge. Using the bacterial test bacteria were identified during only four weeks after challenge. Bvg- B. pertussis avirulent mutants were accumulated for the infection time. The percentage of the avirulent bacteria in the B. pertussis population reached 50% in 7-9 weeks after challenge. The obtained results show that the laboratory mice can be used for study of the B. pertussis insertion mutant formation dynamics in vivo and confirm the hypothesis about insertional bvg- B. pertussis virulent mutants accumulation during development of pertussis infection in human.
Kozyr' AV, Luneva NM, Khlyntseva AE
… +3 more, Shemiakin IG, Krasavtseva ON, Kolesnikov AV
Mol Gen Mikrobiol Virusol
· 2013 · PMID 24645273
The spread of the New Delhi metallo-beta-lactamase (NDM-1), a plasmid-borne enzyme conferring bacterial resistance to any known beta-lactam antibiotics, represents the global health threat. There is an urgent need to dev...The spread of the New Delhi metallo-beta-lactamase (NDM-1), a plasmid-borne enzyme conferring bacterial resistance to any known beta-lactam antibiotics, represents the global health threat. There is an urgent need to develop the efficient NDM-1 inhibitors of various mode of action thereby necessitating structural studies of the enzyme as well as analysis of the secretion pathway and localization of the protein. The recombinant full-length NDM-1 is produced in E. coli in the inactive form and is mostly accumulated in the inclusion bodies. The secreted recombinant NDM-1 forms are several N-terminally truncated species. The robust expression system capable of high-level production of the full-length NDM-1 and derivatives thereof is required to obtain NDM-1 in the quantities necessary for drug discovery, diagnostics, and research purposes. Therefore, we developed a new system that utilizes antibiotic pressure to select E. coli producing increased quantity of soluble NDM-1 and showed that an increase in the NDM-1 solubility occurs in the bacterial clones producing increased amounts in the chaperones.
Pliuta VA, Andreenko IuV, Kuznetsov AE
… +1 more, Khmel' IA
Mol Gen Mikrobiol Virusol
· 2013 · PMID 24645272
In the natural ecosystems, most bacteria exist as specifically organized biofilms attached to various surfaces; the biofilms have a complex architecture and are surrounded by an exopolymeric matrix. The bacteria in the b...In the natural ecosystems, most bacteria exist as specifically organized biofilms attached to various surfaces; the biofilms have a complex architecture and are surrounded by an exopolymeric matrix. The bacteria in the biofilms are extremely resistant to antibacterial agents. The ability of the pathogenic bacteria to produce biofilms causes serious problems in medicine. Therefore, the study of the action of different compounds with antibacterial activity is of great interest. In this work, we studied the effect of the hydrogen peroxide (H2O2) on the formation of biofilms by Pseudomonas aeruginosa PAO1. It was shown that H2O2 in concentrations that do not suppress bacterial growth (or suppress it only weakly) stimulates the formation of the biofilms. At higher concentrations, H2O2 inhibits the formation of the biofilms. In order to determine if the stimulation of the biofilm formation depends on Quorum Sensing (QS) regulation, the plasmid pME6863 containing the heterologous gene aiiA encoding the N-acyl-homoserine lactonase AiiA was introduced into P. aeruginosa PAO1. The synthesis by cells of this enzyme degrading N-acyl-homoserine lactones (AHL), signaling molecules of the QS systems, led to the absence of the stimulation of the biofilm formation by the action of H2O2. This fact indicates that the stimulation of the biofilm formation in the presence of H2O2 depends on the functioning of the QS systems of the gene expression regulation of P. aeruginosa PAO1.