RESUMEN
We analysed clinical and pharmacological factors influencing resistance to clopidogrel in 250 patients with cardiovascular diseases during 18 months. It was shown that the risk ofresistance depends on the form of coronary heart disease, carbohydrate metabolism, the AA genotype of CYP2C19*2 and TBS1 genes. The cardiovascular events significantly morefrequently occurred during 12 and 18 months in resistant diabetics and in the patients with an allele lacking the *2/*3 CYP2C9 gene function and AT/TT polymorphism of the thromboxane synthase gene TBS1.
Asunto(s)
Enfermedades Cardiovasculares/tratamiento farmacológico , Citocromo P-450 CYP2C19/genética , ADN/genética , Resistencia a Medicamentos/genética , Polimorfismo Genético , Tromboxano-A Sintasa/genética , Ticlopidina/análogos & derivados , Alelos , Enfermedades Cardiovasculares/metabolismo , Clopidogrel , Citocromo P-450 CYP2C19/metabolismo , Femenino , Genotipo , Humanos , Masculino , Persona de Mediana Edad , Inhibidores de Agregación Plaquetaria/uso terapéutico , Estudios Retrospectivos , Tromboxano-A Sintasa/metabolismo , Ticlopidina/uso terapéuticoRESUMEN
AIM: To study the characteristics of group A streptococcal infection epidemic process in children aged 12 - 14 years arrived to summer camp "Orlenok" (Tuapse) from different regions of Russia. MATERIALS AND METHODS: Epidemiological (retrospective analysis of incidence of acute respiratory infections, tonsillitis, and scarlet fever), microbiological (isolation and identification of group A streptococci [GAS]), and molecular biological (pulse-electrophoresis, analysis of spe and emm genes) methods were used for the study. Objects of the study were GAS strains isolated from patients and carriers. RESULTS: Performed genotyping showed that cases of GAS infection in newly formed children collectives were caused by 2 - 3 epidemically important clones, which were genotypically heterogenous. CONCLUSION: Performed molecular biologic studies demonstrated polyclonal structure of GAS that determines the features of development of epidemic process.
Asunto(s)
Brotes de Enfermedades , Infecciones Estreptocócicas/epidemiología , Streptococcus pyogenes/clasificación , Adolescente , Antígenos Bacterianos/genética , Proteínas de la Membrana Bacteriana Externa/genética , Proteínas Bacterianas/genética , Proteínas Portadoras/genética , Portador Sano , Niño , Exotoxinas/genética , Humanos , Incidencia , Actividades Recreativas , Epidemiología Molecular , Federación de Rusia/epidemiología , Streptococcus pyogenes/genética , Streptococcus pyogenes/aislamiento & purificaciónRESUMEN
Twenty-eight strains of group A streptococci (GAS) isolated from patients with lacunar and fibrinous-necrotic forms of tonsillitis during its 1st or recurrent episode were tested for presence of genes of erythrogenictoxins A, B, C, and Fusing PCR assay. Obtained results allow to consider that clinical features of the disease (severity, repeatedness, clinical form) can be determined by toxigenicity of GAS. Express identification of S. pyogenes on the basis of detection of erythrogenic toxins genes spe B and spe F (mf) can be used for etiologic confirmation of diagnosis, whereas detection of erythrogenic toxins genes spe A and spe C can be recommended for prediction of the disease's course.
Asunto(s)
Exotoxinas/genética , Infecciones Estreptocócicas/microbiología , Streptococcus pyogenes/patogenicidad , Superantígenos/genética , Tonsilitis/microbiología , Adolescente , Adulto , Proteínas Bacterianas/genética , ADN Bacteriano/análisis , Genes Bacterianos , Humanos , Proteínas de la Membrana/genética , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Infecciones Estreptocócicas/patología , Streptococcus pyogenes/genética , Streptococcus pyogenes/aislamiento & purificación , Tonsilitis/patologíaRESUMEN
The primer systems for the PCR detection of four house-keeping genes of bartonellae in clinical material were developed and tested. The tactics of the species RFLP typing was also developed and tested. The scheme of the species RFLP typing of bartonellae was tested using as an example two strains for the first time isolated in Russia from patients with endocarditis and fever of uncertain origin. The results of the typing were supported by sequencing of the amplicons obtained. According to the sequencing the isolates were attributed to the sub species Bartonella vinsonii, subsp. arupensis. The necessity of molecular epidemiological analysis of bartonelloses in Russia was substantiated.
Asunto(s)
Técnicas de Tipificación Bacteriana , Bartonella/clasificación , Biología Molecular/métodos , Reacción en Cadena de la Polimerasa/métodos , Polimorfismo de Longitud del Fragmento de Restricción , Bartonella/genética , ADN Bacteriano/análisis , ADN Bacteriano/genética , Genes Bacterianos , FilogeniaRESUMEN
Pulse-electrophoresis, sequencing of emm genes coding protein M and PCR analysis of speA, speB, and speC genes were used for characterization of group A streptococci (GAS) isolated in different years in Moscow and Tuapse mostly from children and military staff. It has been shown that epidemic process of streptococcal infection caused by GAS in Moscow is based on circulation of many independent clones of Streptococcus pyogenes. Obtained data on complex typing of S. pyogenes would be useful for study of molecular epidemiology of diseases caused by GAS and improvement of epidemiologic surveillance.
Asunto(s)
Epidemiología Molecular , Infecciones Estreptocócicas/epidemiología , Streptococcus pyogenes/clasificación , Adulto , Antígenos Bacterianos/genética , Proteínas de la Membrana Bacteriana Externa/genética , Proteínas Bacterianas/genética , Proteínas Portadoras/genética , Guarderías Infantiles , Preescolar , Electroforesis en Gel de Campo Pulsado , Exotoxinas/genética , Femenino , Humanos , Masculino , Proteínas de la Membrana/genética , Personal Militar , Federación de Rusia/epidemiología , Streptococcus pyogenes/genética , Superantígenos/genética , Población UrbanaRESUMEN
A total of 103 blood samples collected from wild small mammals captured in the Prioksko-Terrasny Reserve on the south of Moscow region were studied to determine the bartonellae prevalence. The examined species were the yellow-necked mice Apodemus flavicollis (35 samples), the European wood mouse Apodemus uralensis (10 samples), the bank vole Clethrionomys glareolus (51 samples), the house mouse Mus musculus (3 samples), the common vole Microtus arvalis (2 samples), and the shrew Sorex araneus (2 samples). Initially, we obtained 76 bacterial Bartonella-like isolates after plating onto the surface of the solid nutrient media. 66 of them were PCR-positive at least for three of four targets, gltA, ftsZ, ribC and 16S RNA. Thus, the percentage of the infection in the studied community was 64%. Subsequent RFLP assay showed that obtained isolates belonged to the Bartonella grahamii and/or B. taylorii species. In 7 cases we found both bartonellae species in one animal. These data were confirmed by direct sequencing of four ftsZ, four ribC and two gltA amplicons. According to our data, there is no any marked host specificity for these bartonellae species. Now we have laid the bartonellae strain collection consisting of 31 isolates. To our knowledge, this is the first investigation of the bartonellae prevalence in wild small mammals performed in Russia. The comparison of our data with those obtained by European researchers and issues of coinfection by different bartonellae species and host specificity are discussed.
Asunto(s)
Bartonella/genética , Reservorios de Enfermedades/microbiología , Eulipotyphla/microbiología , Muridae/microbiología , Animales , Infecciones por Bartonella/transmisión , Cartilla de ADN , Moscú , Filogenia , Reacción en Cadena de la PolimerasaRESUMEN
The species and strain composition of bifidobacteria in 29 children both sexes, aged 8 to 16 months, was studied. Species-specific primers and PCR were used to determine to which species the predominant strains of bifidobacteria, isolated from feces by cultural methods, belonged. Bifidobacteria were found in 28 (96.5%) children; their number was 10.2 +/- 0.7 ECU per a gram of the material. B. longum and B. bifidum were frequent (71.4 and 53.5%, respectively). The level of quantitative detection used in the study also allowed revealing of B. catenulatum (17.9%) and B. breve (14.4%). A high titer of B. dentium was found in one case (3.6%). B. adolescentis and B. angulatum were not found in any patient. The average number of species found in one child was 1.7 +/- 0.7. RAPD-PCR and investigation of plasmid profile were used to determine possible belonging of the isolates to different strains. The average number of strains per one sample was 2.3 +/- 1.2. Nine unique plasmid bifidobacterial strains were isolated from 7 children. In 3 children the intestinal tract was found to be colonized by both plasmid and non-plasmid-carrying strains of one bifidobacterial species.
Asunto(s)
Bifidobacterium/clasificación , Factores de Edad , Técnicas Bacteriológicas , Bifidobacterium/genética , Bifidobacterium/aislamiento & purificación , Cartilla de ADN , ADN Bacteriano/análisis , Heces/microbiología , Femenino , Humanos , Lactante , Intestinos/microbiología , Masculino , Plásmidos , Reacción en Cadena de la Polimerasa , Factores SexualesRESUMEN
The results of the intraspecific typing of group A streptococci (SGA), isolated from patients with different manifestations of group A streptococcal infection, carried out in Russia for the first time, are presented. The genotypic method of emm-typing, based on sequencing the DNA area coding the variable part of the molecule of SGA M-protein was used. The data obtained in this study made it possible to follow changes in the pattern of SGA M-types in Moscow and the specific features of SGA in comparison with the analogous data on other territories. New emm-types of SGA circulating on the territory of Russia were detected and described.
Asunto(s)
Antígenos Bacterianos/genética , Proteínas de la Membrana Bacteriana Externa/genética , Proteínas Portadoras/genética , Infecciones Estreptocócicas/epidemiología , Streptococcus pyogenes/genética , Antígenos Bacterianos/química , Proteínas de la Membrana Bacteriana Externa/química , Secuencia de Bases , Proteínas Portadoras/química , Genotipo , Humanos , Datos de Secuencia Molecular , Federación de Rusia/epidemiología , Alineación de Secuencia , Infecciones Estreptocócicas/microbiología , Streptococcus pyogenes/aislamiento & purificaciónRESUMEN
To study the structural arrangement of the chromosomal region containing vir genes of Bordetella pertussis the corresponding 15 kb fragment of Bordetella pertussis chromosomal DNA has been cloned. The sequence homology to an earlier characterized Bordetella pertussis genetical element RSBP1 and flanked by two 400 bp inverted repeats has been shown to be located at an end of a BamHI fragment. The restriction map of Bordetella pertussis 475 coincides with the previously published maps of Bordetella pertussis Tohama and 18323 permitting one to conclude the definite conservatism of the cloned sequence. The preliminary data obtained make possible mapping of the RSBP1 homologous sequence adjacent to adenylate cyclase, agglutinin 2 and pertussis toxin genes. The possible role of RSBP1 elements in the regulation of Bordetella virulence is suggested.
Asunto(s)
Bordetella pertussis/genética , Cromosomas Bacterianos , Bordetella pertussis/patogenicidad , Clonación Molecular , ADN Bacteriano/genética , ADN Bacteriano/aislamiento & purificación , Desoxirribonucleasa BamHI/genética , Genes Bacterianos , Hibridación de Ácido Nucleico , Operón , Plásmidos , Mapeo RestrictivoRESUMEN
A number of repeated sequences was identified in the chromosome of Bordetella pertussis by the electron microscopic analysis of the chromosomal DNA of this microorganism. One of the sequences was cloned on the vector plasmid pHC79. It is shown to consist of two elements RSBP1 and RSBP2. The first elements is probably identical to an RS-element described previously. The cloned RSBP1 element is shown to stimulate the deletion formation in the genome of the plasmid pMKII and is able to transpose into the chromosome of Escherichia coli. The latter properties permit one to classify RSBP1 as an element belonging to a class of migrating genetical elements.
Asunto(s)
Bordetella pertussis/genética , Elementos Transponibles de ADN , Cromosomas Bacterianos/ultraestructura , Clonación Molecular , Escherichia coli/genética , Genes Bacterianos , Hibridación de Ácido Nucleico , Plásmidos , Secuencias Repetitivas de Ácidos Nucleicos , Mapeo RestrictivoRESUMEN
Expression of the cloned operon encoding the pertussis toxin synthesis under the control of operons own vir-dependent promoter or vir-independent promoter of Escherichia coli origin was studied. Proteins produced by the recombinant strains have been characterized. The pertussis toxin operon was shown to express under the control of both homologous and heterologous promoters in Bordetella bronchiseptica cells. Use of the lac-promoter increases the yield of produced toxin twofold. Copy number of operon in the cell does not influence the level of toxin production. The synthesized protein can be transported into the culture medium. The biological and physico-chemical properties of the protein are similar to the ones of the natural pertussis toxin. Bordetella bronchiseptica strain producing the toxin with decreased toxic activity has been obtained. Thus, a simple system for cellular expression of the toxin operon was constructed in Bordetella bronchiseptica. It permits one to construct new strains producing nontoxic derivatives of the pertussis toxin for construction of nonreactogenic vaccines.
Asunto(s)
Bordetella bronchiseptica/genética , Operón , Toxina del Pertussis , Regiones Promotoras Genéticas , Factores de Virulencia de Bordetella/genética , Animales , Western Blotting , Células CHO , Cricetinae , Electroforesis en Gel de Poliacrilamida , Expresión Génica , Plásmidos , Recombinación Genética , Factores de Virulencia de Bordetella/biosíntesis , Factores de Virulencia de Bordetella/toxicidadRESUMEN
This work continues a series of reports devoted to the study of a repeated sequence isolated from the Bordetella pertussis chromosome (RSBP--Repeated Sequence of Bordetella Pertussis). The RSBP was earlier demonstrated to have a structure similar to IS elements and exhibit some properties of mobile genetic elements. The results of this work demonstrate the ability of this novel genetic element to stimulate the formation of cointegrates between independent replicons. We have studied the structure of five cointegrates formed by integration of a resident RSBP-containing plasmid into three different sites of a recipient replicon. The studied cointegrates are capable of resolution with the release of a resident plasmid independent of the host cell RecA function.
Asunto(s)
Bordetella pertussis/genética , Cromosomas Bacterianos , Elementos Transponibles de ADN , Escherichia coli/genética , ADN Bacteriano/ultraestructura , Microscopía Electrónica , Hibridación de Ácido Nucleico , Plásmidos , Secuencias Repetitivas de Ácidos Nucleicos , Mapeo RestrictivoRESUMEN
Mutants of Salmonella typhimurium with an impaired process of transition to the nonculturable state were tainted. Mutants were divided into four phenotypic groups. In four mutants (representatives of each phenotypic group), genes with TnPhoA transposon insertions were cloned; these insertions caused a disturbance in the process of mutant cell transition to the nonculturable state. Nucleotide sequences of mutant gene fragments were determined. Comparison of nucleotide sequences obtained with a data bank on DNA nucleotide sequences of enterobacterial genomes allowed the identification of four genes involved in the control of nonculturable form generation in salmonellae.
Asunto(s)
Bases de Datos Factuales , Escherichia coli/genética , Genes Bacterianos , Salmonella typhimurium/genética , Secuencia de Bases , Clonación Molecular , Elementos Transponibles de ADN , Datos de Secuencia Molecular , Mutación , Fenotipo , Salmonella typhimurium/crecimiento & desarrollo , Homología de Secuencia de Ácido NucleicoRESUMEN
The repeated sequence from Bordetella pertussis chromosome was cloned using the method described by Ohtsubo. The sequences of the characterized B. pertussis chromosome are homologous to the previously described sequences and analogous in the structure to the already known IS elements. The parental recombinant plasmids containing the RS were unstable and segregated to the plasmids of different structure. The segregants' structure is characterized in this paper. It is shown in our study that the RS element is able to stimulate intragenomic rearrangements, such as deletions. It is shown that at least one deletion begins precisely after 3' end of RS and terminates with the sequence which is completely homologous to ten terminal nucleotides of this one. Probably, RSs stimulate at high frequency the formation of deletions, which appear to be the result of recA-independent site-specific recombination between short direct repeats.
Asunto(s)
Bordetella pertussis/genética , Inversión Cromosómica , Cromosomas Bacterianos , Secuencias Repetitivas de Ácidos Nucleicos , Secuencia de Bases , Clonación Molecular , ADN Bacteriano/genética , ADN Bacteriano/ultraestructura , Microscopía Electrónica , Datos de Secuencia Molecular , Plásmidos , Rec A Recombinasas/metabolismo , Recombinación Genética , Mapeo RestrictivoRESUMEN
The 2.3 kb BamHI-EcoRI subclone has been isolated and sequenced from the 15 kb BamHI chromosomal fragment of Bordetella pertussis comprising also the vir gene. The sequence contained one copy of a transposon-like structure, very similar to the RSs of B. pertussis recently characterized, the unique sequence of B. pertussis chromosomal DNA and an inverted sequence complementary to 402 bp of 3' end of the B. pertussis RS element. The fragment of plasmid pUC4K containing the gene for kanamycin resistance (Km) was integrated in the XhoI site of the unique portion of the transposon-like structure of plasmid DNA (Ap(r), Km(r)). The clones in amount of 2% tested had the Ap-S phenotype. The recombinant plasmid from the Ap(s) phenotype clones lost one of the repeats and a portion of the gene bla as a result of deletion. The conclusion is drawn that RSs of B. pertussis stimulate the formation of deletions.
Asunto(s)
Bordetella pertussis/genética , Cromosomas Bacterianos , Elementos Transponibles de ADN , Reordenamiento Génico , Resistencia a la Ampicilina/genética , Secuencia de Bases , Clonación Molecular , Escherichia coli/genética , Genes Bacterianos , Datos de Secuencia Molecular , Secuencias Repetitivas de Ácidos NucleicosRESUMEN
The diagnostic value of preparations is commonly characterized by sensitivity and specificity. But not all these characteristics make it possible to decide unequivocally which of the preparations to be compared is superior to the other one with respect to its diagnostic value. It is proposed that in the choice of a diagnostic preparation its capacity to provide data for exact diagnosis should be considered, i.e. the additional characteristic indicating the spread of the disease under study. As an example, the comparison of the diagnostic value of conventional methods and the polymerase chain reaction in the diagnostics of helicobacteriosis is presented. The described method for the evaluation of the diagnostic value of the preparation is well-grounded, simple and obvious.
Asunto(s)
Infecciones por Helicobacter/diagnóstico , Helicobacter pylori/aislamiento & purificación , Reacción en Cadena de la Polimerasa/métodos , Juego de Reactivos para Diagnóstico , Antígenos Bacterianos/genética , Proteínas Bacterianas/genética , Cartilla de ADN , Genes Bacterianos , Helicobacter pylori/genética , Humanos , Sensibilidad y EspecificidadAsunto(s)
Infecciones por Bordetella/diagnóstico , Bordetella pertussis/genética , Bordetella/genética , ADN Bacteriano/análisis , Infecciones Estreptocócicas/diagnóstico , Streptococcus/genética , Tos Ferina/diagnóstico , Adulto , Niño , Diagnóstico Diferencial , Humanos , Lactante , Recién Nacido , Reacción en Cadena de la PolimerasaRESUMEN
There is the HP diagnostic methods review. The classification of main methods is presented. Diagnostic significance and defects of all these methods were demonstrated. There are own data of HP and its strains revealing in the patients with duodenal peptic ulcer.