RESUMO
The transcription factor glioma-associated oncogene homolog 1 (GLI1) has a central function in gastrointestinal tract development and homeostasis. A non-synonymous single-nucleotide polymorphism (SNP) (rs2228226; Q1100E) in GLI1, which impairs GLI1 function in vitro, has been proposed as a risk factor for inflammatory bowel disease (IBD). In this study, we assessed the cumulative evidence for association of GLI1 with IBD. New genotype data for rs2228226 from New Zealand (907 controls, 990 IBD patients) and Belgian Caucasian case-control data sets (312 controls, 1214 IBD patients) were combined with data from the National Institute of Diabetes and Digestive and Kidney Diseases and three previously studied Caucasian case-control data sets. Meta-analysis of rs2228226 did not detect any association with ulcerative colitis (UC) (P=0.09, odds ratio (OR)=1.07, 95% confidence interval (CI)=0.92-1.24), Crohn's disease (CD) (P=0.29, OR=1.06, 95% CI=0.93-1.21) or overall IBD (P=0.15, OR=1.05, 95% CI=0.92-1.19). Our analyses of rs2228226 suggest that GLI1 is not a significant risk factor for IBD in Caucasians.
Assuntos
Predisposição Genética para Doença/genética , Doenças Inflamatórias Intestinais/genética , Fatores de Transcrição/genética , População Branca/genética , Adolescente , Adulto , Idoso , Estudos de Casos e Controles , Feminino , Humanos , Doenças Inflamatórias Intestinais/epidemiologia , Masculino , Pessoa de Meia-Idade , Fatores de Risco , Adulto Jovem , Proteína GLI1 em Dedos de ZincoRESUMO
Streptococcus suis is an important pathogen of pigs causing arthritis, pneumonia and meningitis and is an occupational disease of farmers and those in the meat industry. As with other streptococci, both virulent and avirulent strains of S. suis are frequently carried asymptomatically in the tonsillar crypts and nasal cavities. Little is known about the process by which virulent strains cross the mucosal epithelia to generate systemic disease and whether this process requires expression of specific bacterial virulence factors. Although putative virulence factors have been postulated, no specific role in the disease process has yet been demonstrated for these factors. This study is the first demonstration that virulent strains of S. suis both invade and lyse HEp-2 cells, a continuous laryngeal epithelial cell line, and that at least one bacterial virulence factor, suilysin, is involved in this process.
Assuntos
Células Epiteliais/microbiologia , Proteínas Hemolisinas/fisiologia , Streptococcus suis/patogenicidade , Anticorpos Monoclonais/imunologia , Anticorpos Monoclonais/farmacologia , Meios de Cultivo Condicionados/farmacologia , DNA Bacteriano/genética , Relação Dose-Resposta a Droga , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/ultraestrutura , Proteínas Hemolisinas/genética , Proteínas Hemolisinas/imunologia , Humanos , Compostos Orgânicos , Reação em Cadeia da Polimerase , Especificidade da Espécie , Streptococcus suis/efeitos dos fármacos , Streptococcus suis/ultraestrutura , Células Tumorais Cultivadas , Virulência/efeitos dos fármacosRESUMO
Species-specific oligonucleotide probes and a universal oligonucleotide probe derived from sequences of 16S rRNA were hybridised to chromosomal DNA from Streptococcus agalactiae, S. dysgalactiae, S. parauberis and S. uberis following digestion with EcoRI. Due to the presence of a unique EcoRI site in each 16S rRNA gene, the number of hybridised fragments was indicative of the number of 16S rRNA genes. Southern hybridisation indicated six 16S rRNA genes in ten isolates of S. agalactiae, five genes in ten isolates of S. uberis, five genes in six isolates and six in another isolate of S. dysgalactiae, and six genes in four isolates of S. parauberis. For a fifth isolate of S. parauberis, six 16S rRNA genes were indicated by the universal probe but only five when hybridised to the species-specific probe, indicating sequence variation (microheterogeneity) within the probe target region.
Assuntos
Dosagem de Genes , RNA Ribossômico 16S/genética , Streptococcus/genética , Sequência de Bases , Southern Blotting , DNA Bacteriano/análise , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , Streptococcus agalactiae/genéticaRESUMO
16S rRNA of Streptococcus agalactiae, S. uberis, and S. parauberis was bound to streptavidin-coated magnetic beads by using a biotinylated oligonucleotide probe complementary to a highly conserved region of the molecule. In-solution hybridization of radiolabelled oligonucleotide probes to immobilized 16S rRNA allowed the specific identification of S. agalactiae and S. parauberis but not S. uberis. PCR was used to amplify a species-specific region of the 16S rRNA gene from these species. One of the PCR primers was biotinylated at the 5' end to allow purification of the amplified product on streptavidin-coated magnetic beads and subsequent denaturation to yield immobilized single-stranded DNA. Radiolabelled oligonucleotide probes were hybridized in solution to the single-stranded target molecule and enabled species-specific identification of the target organism. This protocol overcame problems associated with hybridization of the S. uberis-specific probe to 16S rRNA in solution. A similar procedure may enable the specific detection of other streptococci which exhibit a species-specific sequence in this region of the gene.
Assuntos
Reação em Cadeia da Polimerase/métodos , Streptococcus/genética , Animais , Sequência de Bases , Bovinos , DNA Bacteriano/genética , DNA Ribossômico/genética , Estudos de Avaliação como Assunto , Feminino , Dados de Sequência Molecular , Conformação de Ácido Nucleico , Hibridização de Ácido Nucleico , Sondas de Oligonucleotídeos/genética , RNA Bacteriano/química , RNA Bacteriano/genética , RNA Ribossômico 16S/química , RNA Ribossômico 16S/genética , Homologia de Sequência do Ácido Nucleico , Especificidade da Espécie , Streptococcus/classificação , Streptococcus/isolamento & purificação , Streptococcus agalactiae/genética , Streptococcus agalactiae/isolamento & purificaçãoRESUMO
Genes encoding 16S rRNA were sequenced from 16 species of Staphylococcus. Sequence analysis highlighted a potential Staph. aureus-specific region and a complementary oligonucleotide probe was synthesized and its specificity tested. Northern blotting indicated molecular specificity, and dot blots to RNA from Staph. aureus, Staph. capitis, Staph. caprae, Staph. carnosus, Staph. caseolyticus, Staph. cohnii, Staph. epidermidis, Staph. gallinarum, Staph. haemolyticus, Staph. hominis, Staph. hyicus, Staph. saprophyticus, Staph. sciuri, Staph. simulans, Staph. warneri and Staph. xylosus indicated species-specificity.
Assuntos
Sondas de Oligonucleotídeos , RNA Bacteriano/genética , RNA Ribossômico 16S/genética , Staphylococcus aureus/genética , Composição de Bases , Sequência de Bases , Northern Blotting , Genes Bacterianos , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , Sondas de Oligonucleotídeos/química , RNA Bacteriano/química , RNA Ribossômico 16S/isolamento & purificação , Alinhamento de Sequência , Homologia de Sequência do Ácido Nucleico , Especificidade da Espécie , Staphylococcus/genética , Staphylococcus aureus/isolamento & purificaçãoRESUMO
Oligonucleotide probes specific for 16S rRNA and capable of differentiating Streptococcus uberis and S. parauberis from each other and other esculin-hydrolyzing streptococci were developed. Use of a mini-RNA extraction technique for gram-positive cocci associated with bovine mastitis has allowed the probes to be used for identification of esculin-hydrolyzing streptococci from two dairy herds at the Institute for Animal Health, Compton, United Kingdom. One hundred seventy-nine of 206 isolates were identified as S. uberis, 3 were identified as S. parauberis, and 24 were not identified. Isolates not identified by the probes were tested biochemically and found to be mainly Enterococcus faecium, E. faecalis, or S. bovis.
Assuntos
Técnicas de Tipagem Bacteriana , Mastite Bovina/microbiologia , Sondas de Oligonucleotídeos , RNA Ribossômico 16S/genética , Streptococcus/classificação , Animais , Sequência de Bases , Northern Blotting , Bovinos , Esculina/metabolismo , Feminino , Dados de Sequência Molecular , Especificidade da Espécie , Streptococcus/genética , Streptococcus/isolamento & purificaçãoRESUMO
The partial 16S rRNA sequences of 24 Streptococcus species were determined by reverse transcription. A comparative analysis of these sequences and the sequences of seven previously studied streptococcal species revealed the presence of several clusters within the genus. The clusters obtained from the sequence analysis agreed in general with the groups outlined on the basis of the results of nucleic acid hybridization studies, but there were some exceptions. The pyogenic group was extended to include Streptococcus agalactiae, S. parauberis, S. porcinus, and S. uberis. Four oral groups were discerned; these four groups centered on S. mutans, S. salivarius, S. anginosus, and S. oralis. Some species (e.g., S. suis and S. acidominimus) did not cluster with any particular group. Our findings are discussed in the context of data from other genetic and chemotaxonomic studies.