RESUMO
The human leukocyte antigen (HLA) system has a major role in the regulation of the immune response as it is involved in the defense against pathogens. Some studies have reported that HLA class II genes play a strong role in severe cases of pulmonary tuberculosis (PTB) in several populations. Thus the aim of the study was to compare the HLA-class II alleles of patients with drug resistant tuberculosis with those of healthy controls from the same ethnic group in Kazakhstan. The aim of the present study was to evaluate the correlation of HLA-class II alleles by patients with drug resistant tuberculosis and the healthy controls of the same ethnic group in Kazakhstan. The HLA-class II alleles of 76 patients with tuberculosis (TB) and 157 healthy volunteers were investigated using sequence-based typing (SBT)-method. HLA-DQA1*03:02 HLA-DRB1*08:01 and DRB1*08:03 occurred more frequently (P = 0.05) in patients with drug resistant tuberculosis than in controls. We observed a possible association between certain HLA alleles and TB that are specific for the Kazakh population. Further studies are needed to confirm our findings using a larger number of patients with drug resistant tuberculosis.
Assuntos
Predisposição Genética para Doença , Cadeias alfa de HLA-DQ/genética , Cadeias HLA-DRB1/genética , Haplótipos , Tuberculose Resistente a Múltiplos Medicamentos/genética , Tuberculose Pulmonar/genética , Adulto , Antituberculosos/uso terapêutico , Estudos de Casos e Controles , Feminino , Frequência do Gene , Cadeias alfa de HLA-DQ/classificação , Cadeias alfa de HLA-DQ/imunologia , Cadeias HLA-DRB1/classificação , Cadeias HLA-DRB1/imunologia , Teste de Histocompatibilidade , Humanos , Cazaquistão , Masculino , Pessoa de Meia-Idade , Mycobacterium tuberculosis/efeitos dos fármacos , Mycobacterium tuberculosis/imunologia , Tuberculose Resistente a Múltiplos Medicamentos/tratamento farmacológico , Tuberculose Resistente a Múltiplos Medicamentos/imunologia , Tuberculose Resistente a Múltiplos Medicamentos/microbiologia , Tuberculose Pulmonar/tratamento farmacológico , Tuberculose Pulmonar/imunologia , Tuberculose Pulmonar/microbiologiaRESUMO
BKV reactivation is associated with impaired graft function in kidney transplant patients. The objective of our study was to determine the prevalence of BKV infection in consecutive pediatric kidney transplant recipients at our center. Fifty-eight pediatric kidney transplant recipients were studied. The mean age at screening was 9.4 ± 2.8 yr, and samples were obtained at a median of 2.4 ± 1.4 yr after transplantation. BKV-DNA was analyzed in urine and plasma by quantitative PCR. Occurrences of BK-DNAuria and BK-DNAemia did not change in the first two yr after transplantation in children and amounted to 21-23% and 7-8%, respectively (p > 0.05). In the third year, the occurrences of BK-DNAuria and BK-DNAemia increased insignificantly to 27% and 9% in the pediatric patients. We also determined the subtypes and subgroups of BK virus isolated from Russian renal transplant recipients and found that BKV isolates were composed of subtypes Ib-2 and IV/c2. The data we obtained indicate that although only 5% of BKVAN cases occurred between years two and five post-transplantation, it seems necessary to regularly monitor pediatric patients for BKV infection through the third year after transplantation.
Assuntos
Vírus BK/isolamento & purificação , Transplante de Rim , Infecções por Polyomavirus/epidemiologia , Complicações Pós-Operatórias/epidemiologia , Infecções Tumorais por Vírus/epidemiologia , Adulto , Vírus BK/classificação , Vírus BK/genética , Sequência de Bases , Estudos de Casos e Controles , Criança , DNA Viral/análise , Feminino , Genótipo , Humanos , Masculino , Dados de Sequência Molecular , Filogenia , Infecções por Polyomavirus/diagnóstico , Infecções por Polyomavirus/etiologia , Infecções por Polyomavirus/virologia , Complicações Pós-Operatórias/diagnóstico , Complicações Pós-Operatórias/virologia , Reação em Cadeia da Polimerase em Tempo Real , Estudos Retrospectivos , Federação Russa , Infecções Tumorais por Vírus/diagnóstico , Infecções Tumorais por Vírus/etiologia , Infecções Tumorais por Vírus/virologia , Carga ViralAssuntos
Proteínas de Bactérias/genética , Chaperonina 60/genética , Produtos Fermentados do Leite/microbiologia , Lactobacillus/classificação , RNA Ribossômico 16S/genética , Técnicas de Tipagem Bacteriana , Chaperonina 60/classificação , Genótipo , Cazaquistão , Lactobacillus/genética , Lactobacillus/isolamento & purificação , Fenótipo , Filogenia , RNA Ribossômico 16S/classificação , Federação Russa , Análise de Sequência de DNARESUMO
Eight novel alleles, two HLA-A and six HLA-B, are described in Kazakh individuals.
Assuntos
Antígenos HLA-A/genética , Antígenos HLA-B/genética , Alelos , Humanos , Cazaquistão , Análise de Sequência de DNARESUMO
Seven novel human leukocyte antigen (HLA)-DRB1 alleles were identified during routine sequence-based typing (SBT) of healthy Kazakh individuals.
Assuntos
Antígenos HLA-DR/genética , Alelos , Sequência de Bases , Análise Mutacional de DNA/métodos , Cadeias HLA-DRB1 , Saúde , Humanos , Cazaquistão , Polimorfismo Genético/fisiologia , Análise de Sequência de DNA/métodosRESUMO
Recent studies of cytochrome P-450 gene polymorphism showed that it may influence the outcome of treatment of Helicobacter pylori (Hp) infection. The literature data concerning effects of genetic factors on the results of eradication therapy of peptic ulcer in the Russian population are scarce and the problem needs further investigation. The aim of this work was to examine CYP2C19 polymorphism in a mixed population of the city and region of Moscow and evaluate its effect on the efficiency of eradication therapy of Hp infection. The prospective study of a cohort of 82 patients with Hp-induced gastric and duodenal ulcers examined in conformity with the current health care standards. All the patients received first line eradication therapy during 7 days. Its efficiency was found to be dependent on CYP2C19 polymorphism among other factors.
Assuntos
Hidrocarboneto de Aril Hidroxilases/genética , DNA Bacteriano/genética , Úlcera Duodenal/genética , Infecções por Helicobacter/genética , Helicobacter pylori/genética , Polimorfismo Genético , Úlcera Gástrica/genética , Adulto , Biópsia , Citocromo P-450 CYP2C19 , Sistema Enzimático do Citocromo P-450/genética , Úlcera Duodenal/tratamento farmacológico , Úlcera Duodenal/microbiologia , Endoscopia Gastrointestinal , Feminino , Seguimentos , Infecções por Helicobacter/tratamento farmacológico , Infecções por Helicobacter/microbiologia , Helicobacter pylori/efeitos dos fármacos , Helicobacter pylori/isolamento & purificação , Humanos , Mucosa Intestinal/microbiologia , Mucosa Intestinal/patologia , Masculino , Estudos Prospectivos , Úlcera Gástrica/tratamento farmacológico , Úlcera Gástrica/microbiologiaRESUMO
A high-throughput method based on the minisequencing reaction followed by MALDI-TOF mass spectrometry has been developed for detecting C-->T transitions in 291 methylation sites of M.Hpy99XI Helicobacter pylori J99. The study has shown an absolute (100%) accuracy of the new method.
Assuntos
Metilação de DNA/genética , DNA Bacteriano/genética , Helicobacter pylori/genética , Mutação Puntual , Análise Mutacional de DNA/métodos , Sensibilidade e EspecificidadeRESUMO
The frequence of mutations in the rifampicin resistant (RIF(r)) clones of microorganisms after adaption to ofloxacin and metronidazole was investigated to estimate the biological cost of H. pylori rifampicin (RIF) resistance. Mutations in rpoB gene responsible for RIF resistance of H. pylori were shown to have biological cost and be compensated by additional mutations in the microorganism genome. Comparison of the mutation frequency in the presence of metroniazole demonstrated that the acquired resistance to RIF resulted in changing of the adaptative capacity of the RIF(r) clones of H. pylori to metronidazole. Thus, a significant increase of the mutation frequency (> 700 times) in one of the RIF(r) clones and a broad spectrum of the mutations responsible for resistance to metronidazole vs. the H. pylori initial strain 26695 were observed. The findings could be evident of the fact that the adaptation to RIF changed the properties of the cell on one hand in such a way that its mutation capacity increased and that the target selection on the other hand revealed hypermutable cells, likely usual for the bacterial population.
Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana/efeitos dos fármacos , Helicobacter pylori/efeitos dos fármacos , Adaptação Biológica , Proteínas de Bactérias/genética , RNA Polimerases Dirigidas por DNA/genética , Genes Bacterianos , Helicobacter pylori/genética , Metronidazol/farmacologia , Mutação , Ofloxacino/farmacologiaAssuntos
Hidrocarboneto de Aril Hidroxilases/genética , Infecções por Helicobacter/tratamento farmacológico , Helicobacter pylori/efeitos dos fármacos , Polimorfismo de Nucleotídeo Único , Antibacterianos/farmacocinética , Antibacterianos/uso terapêutico , Hidrocarboneto de Aril Hidroxilases/fisiologia , Citocromo P-450 CYP2C19 , Frequência do Gene , Infecções por Helicobacter/enzimologia , Humanos , Taxa de Depuração Metabólica , Inibidores da Bomba de Prótons/farmacocinética , Inibidores da Bomba de Prótons/uso terapêutico , Resultado do TratamentoRESUMO
Analysis of 16S ribosomal RNA gene was carried out in 2 DNA samples isolated from vaginal epithelial smears from women suffering from bacterial vaginosis. The composition of vaginal epithelial microbiocenosis in bacterial vaginosis was determined and its significant difference from normocenosis was shown. A laboratory protocol for identification of vaginal epithelial microflora was developed on the basis of 16S ribosomal RNA gene analysis.
Assuntos
Epitélio/microbiologia , RNA Ribossômico 16S/genética , Vagina , Bactérias/genética , Feminino , Humanos , Vagina/anatomia & histologia , Vagina/microbiologia , Vaginose Bacteriana/microbiologiaRESUMO
Comparison of open-reading frames (ORFs) H. pylori 26695 and J99 strains has been revealed prevalence of nucleotide replacements as transitions (more than 3%) above transversions (less than 1%). Prevalence of nucleotide transitions is caused by high speed of C : G to T : A transitions in a coding strand of DNA (3.5-5.3%) and not coding strand (2.9-3.9%). The correspondence rate of transversion (A --> C, A --> T, C --> A, C --> G, G --> C, G --> T, T --> A and T --> G) did not exceed 0.84%. The highest correspondence frequency between C and T was detected in ACGT-ATGT (28.3%) - the site of methylation by active methyltransferase M.Hpy99XI in H. pylori 26695 and J99. Thus one can speculate that predominant transition taking place in H. pylori is mutation of C into T, which is realized through cytosine methylation-deamination mechanism.
Assuntos
Metilação de DNA , Genoma Bacteriano/genética , Helicobacter pylori/genética , Fases de Leitura Aberta/genética , Mutação Puntual , Proteínas de Bactérias/genética , DNA Metiltransferases Sítio Específica (Adenina-Específica)/genética , Especificidade da EspécieRESUMO
To detect point mutations A2115C, A2143G/C, and A2143G in the 23S rRNA gene of Helicobacter pylori associated with resistance of the microorganism to clarithromycin, a new powerful way of analysis was used. This method involved the reaction of minisequencing followed by MALDI-TOF mass spectrometry of reaction products. In ten analyzed clarithromycin-resistant clinical isolates of H. pylori obtained in Russia, the resistance was found to be mediated only by mutation A2144G in the 23S rRNA gene.
Assuntos
Antibacterianos , Claritromicina , Farmacorresistência Bacteriana/genética , Helicobacter pylori/genética , Mutação Puntual , RNA Bacteriano/genética , RNA Ribossômico 23S/genética , Antibacterianos/farmacologia , Claritromicina/farmacologia , Genes Bacterianos/genética , Genes de RNAr/genética , HumanosRESUMO
Using multilocus sequence typing (MLST), 22 Helicobacter pylori isolates from Russia have been characterized. All of the Russian strains were assigned to a single population, hpEurope.
Assuntos
Genes Bacterianos/genética , Helicobacter pylori/genética , Filogenia , Genética Populacional , Humanos , Federação Russa , Análise de Sequência de DNARESUMO
The transcription profiles of four Helicobacter pylori clinical isolates (two cag-negative and two cag-positive) were compared in stationary growth phase using a cDNA-macroarray. The correlation coefficient value between total transcription profiles of clinical isolates H. pylori varied from 0.70 to 0.83. For 44 groups of genes (total number 66) belonging to various functional classes of H. pylori, the correlation coefficient value between these isolates exceeded 0.7, and for 14 groups the value exceeded 0.9. These groups included genes encoding components involved in cell division, adaptations to atypical conditions, electron transport, salvage of nucleosides and nucleotides, glycolysis/gluconeogenesis, folding and stabilization of proteins, translation factors, anaerobic metabolism, and amino acids and amine metabolism. Expression of 52 genes significantly differed between H. pylori clinical isolates. Some of these genes determine microorganism virulence. They include: cytotoxin-associated gene (cagA), genes encoding neutrophil-activating protein (napA), major flagellar protein (flaA), and vacuolizing cytotoxin (vacA), some genes encoding outer membrane proteins (omp), urease alpha and beta subunits (ureA and ureB), and some regulatory proteins, and genes encoding stress-related proteins, such as the chaperone and heat shock protein genes (groEL and dnaK).
Assuntos
Genes Bacterianos , Helicobacter pylori/genética , Transcrição Gênica , Adesinas Bacterianas/genética , Antígenos de Bactérias/genética , Proteínas de Bactérias/genética , Helicobacter pylori/isolamento & purificação , Humanos , Análise de Sequência com Séries de OligonucleotídeosRESUMO
Complete nucleotide cDNA sequence (29715 nucleotides) of SARS-associated coronavirus (strain SoD) isolated for the first time in the territory of the Russian Federation was determined. Phylogenetic analysis revealed maximum similarity between strain SoD genome and Frankfurt 1 strain genome. Three nucleotide substitutions determining two amino acid substitutions were detected.
Assuntos
Genoma Viral , Coronavírus Relacionado à Síndrome Respiratória Aguda Grave/genética , Coronavírus Relacionado à Síndrome Respiratória Aguda Grave/isolamento & purificação , Substituição de Aminoácidos , DNA Complementar/genética , DNA Viral/genética , Humanos , Filogenia , Reação em Cadeia da Polimerase , Federação Russa , Coronavírus Relacionado à Síndrome Respiratória Aguda Grave/classificação , Síndrome Respiratória Aguda Grave/virologiaRESUMO
The goal of the present work was to compare different techniques of molecular typing using as an example clinical isolates of Helicobacter pylori obtained from patients in different regions of Russia. DNA-macroarray genome scanning using individual genes was employed to set up our basic classification of isolates that did or did not contain pathogenicity islands. In parallel, DNA of the same isolates was used in the conventional random amplified polymorphic DNA (RAPD) PCR analysis, and the isolates were also genotyped (cagA, vacA, iceA, and babA status) and their proteomic maps were obtained by means of unidimensional SDS polyacrylamide gel electrophoresis (1D-SDS-PAGE) coupled with identification using peptide mass fingerprinting by MALDI-TOF mass spectrometry. A statistically significant correlation (coefficient of correlation r = 0.25, p = 0.005) was observed between the results of genome scanning and 1D-SDS-PAGE. No correlation was found between RAPD-PCR typing and genome scanning.
Assuntos
Técnicas de Tipagem Bacteriana/métodos , Helicobacter pylori/classificação , DNA Bacteriano/análise , Eletroforese em Gel de Poliacrilamida , Ilhas Genômicas/genética , Helicobacter pylori/isolamento & purificação , Humanos , Técnicas de Sonda Molecular , Reação em Cadeia da Polimerase , Proteômica , Técnica de Amplificação ao Acaso de DNA Polimórfico , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por MatrizRESUMO
DNA macroarrays were used to characterize 17 Helicobacter pylori strains isolated in four geographic regions of Russia (Moscow, St. Petersburg, Kazan, and Novosibirsk). Of all genes, 1272 (81%) proved to occur in all strains and to constitute a functional core of the genome, and 293 (18.7%) were strain-specific and greatly varied among the H. pylori strains. Most (71%) of the latter had unknown functions; the remainder included restriction-modification genes (3-9%), transposition genes (2-4%), and genes coding for outer membrane proteins (2-4%). The Russian H. pylori strains did not differ in genome organization or in the number and distribution of strain-specific genes from strains isolated in other countries.
Assuntos
Variação Genética , Genoma Bacteriano , Helicobacter pylori/genética , Proteínas de Bactérias/genética , Ordem dos Genes , Genes Bacterianos , Helicobacter pylori/patogenicidade , Glicoproteínas de Membrana/genética , Família Multigênica , Análise de Sequência com Séries de Oligonucleotídeos , Fases de Leitura Aberta , Federação Russa , Especificidade da Espécie , Virulência/genéticaRESUMO
A Mycoplasma hominis chromosomal fragment containing the full-length ftsZ gene was cloned and sequenced. Natural expression of this gene was demonstrated by reverse transcription-polymerase chain reaction (RT-PCR) with total RNA. The M. hominis FtsZ protein was shown to differ substantially from its counterparts of two other Mycoplasma species, M. genitalium and M. pneumoniae. The possibility of M. hominis ftsZ expression in Escherichia coli was demonstrated with several bacterial strains. The M. hominis FtsZ protein was isolated from E. coli cells transformed with recombinant plasmids carrying the M. hominis ftsZ gene. Complementation between the E. coli and M. hominis FtsZ proteins was observed in transformants.