RESUMO
We examined the genetic characteristics of a human oral treponeme isolated from subgingival plaque of a patient with periodontal disease that could not be assigned to any of the previously described oral Treponema species. The guanine-plus-cytosine content of its DNA was 51 mole%. The levels of DNA-DNA relatedness of the organism to other Treponema species, including Treponema denticola, Treponema vincentii, Treponema socranskii, Treponema pallidum and Treponema phagedenis, were less than 30%. This study clearly demonstrated that the isolated treponeme could be completely distinct from the established oral Treponema species genetically. Also, the study indicated remarkable genetic heterogeneity among human oral Treponema species.
Assuntos
Placa Dentária/microbiologia , Treponema/genética , Citosina/análise , DNA Bacteriano/química , Heterogeneidade Genética , Guanina/análise , Humanos , Masculino , Pessoa de Meia-Idade , Hibridização de Ácido Nucleico , Periodontite/microbiologia , Homologia de Sequência do Ácido Nucleico , Treponema/classificação , Treponema/isolamento & purificaçãoRESUMO
A new Treponema species, for which we propose the name Treponema medium, was isolated from subgingival plaque from an adult with periodontal disease. The morphological characteristics, differential biochemical characteristics, and protein profiles on sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels of this organism are described. The guanine-plus-cytosine content of the DNA of T. medium is 51 mol%. The levels of DNA-DNA relatedness of the new species to other Treponema species, including Treponema denticola, Treponema vincentii, Treponema socranskii, Treponema pallidum, and Treponema phagedenis, are less than 30%. A phylogenetic analysis based on 16S rRNA sequences distinguished the new Treponema strain from strains belonging to previously described Treponema species. The type strain of T. medium is strain G7201.
Assuntos
Treponema/classificação , Proteínas de Bactérias/análise , Técnicas Bacteriológicas , Composição de Bases , Western Blotting , Meios de Cultura/metabolismo , DNA Bacteriano/análise , Eletroforese em Gel de Poliacrilamida , Microscopia Eletrônica , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , Filogenia , RNA Ribossômico 16S/genética , Homologia de Sequência do Ácido Nucleico , Treponema/genética , Treponema/imunologia , Treponema/ultraestruturaRESUMO
The ability of spirochetes to adhere to collagens was compared among three species of human oral treponemes. Immunoblot analysis demonstrated that type I-, IV-, and V-collagen-binding polypeptides (CBPs) were detected in the heated and unheated preparations from both Treponema denticola ATCC 33520 and T. socranskii subsp. buccale ATCC 35534. Few CBPs, however, were detected in the heated and unheated preparations from a recently characterized isolate, T. medium strain G7201. Immunoelectron microscopy using rabbit antisera against the CBPs from the unheated preparations demonstrated that four CBPs, a 27 kDa type V-CBP of T. denticola ATCC 33520, a 95 kDa type IV-CBP and a 110 kDa type I-CBP of T. socranskii subsp. buccale ATCC 35534, and a 95 kDa type IV-CBP of T. medium strain G7201, were located on the outer envelopes of the individual cells. The adherence of T. denticola to the collagen-coated surfaces was significantly greater than that of T. medium, suggesting that the CBPs on the oral spirochetal cells play an important role in their adherence to collagen-rich connective tissues of the host.
Assuntos
Aderência Bacteriana , Proteínas da Membrana Bacteriana Externa/análise , Proteínas de Transporte/análise , Colágeno/metabolismo , Treponema/química , Treponema/fisiologia , Proteínas da Membrana Bacteriana Externa/imunologia , Proteínas da Membrana Bacteriana Externa/isolamento & purificação , Proteínas da Membrana Bacteriana Externa/metabolismo , Proteínas de Transporte/metabolismo , Eletroforese em Gel de Poliacrilamida , Temperatura Alta , Humanos , Immunoblotting , Microscopia Eletrônica , Peptídeos/análise , Ligação Proteica , Treponema/imunologiaRESUMO
Agar diffusion analysis demonstrated that rat cystatin S, a cysteine proteinase inhibitor, inhibited the growth of all tested strains of a human oral, Gram-negative anaerobic periodontopathogen Porphyromonas gingivalis. Its specific inhibitory activity against this tissue-invasive bacterium but not against other tested oral bacterial species emphasized the importance of specific cysteine proteinases for growth of P. gingivalis.
Assuntos
Cistatinas/farmacologia , Porphyromonas gingivalis/efeitos dos fármacos , Animais , Fusobacterium nucleatum/efeitos dos fármacos , Inibidores do Crescimento/farmacologia , Porphyromonas gingivalis/crescimento & desenvolvimento , Prevotella intermedia/efeitos dos fármacos , Ratos , Cistatinas SalivaresRESUMO
In this study, we demonstrate that Porphyromonas gingivalis fimbrillin, a major component of bacterial fimbriae, is one of the fibronectin-binding proteins and that fibronectin is a potent inhibitor of the adherence of the bacteria to host cells and of the pathogenesis of the bacterium that acts by binding to the fimbriae. A Western blotting (immunoblotting) assay showed that fibronectin binds strongly to P. gingivalis fimbrillin. The fimbrial binding to fibronectin was also evidenced by a binding assay involving 125I-labeled fimbriae. Furthermore, fibronectin markedly inhibited the fimbria-induced expression of interleukin-1beta and neutrophil-specific chemoattractant KC genes in macrophages. The inhibitory action depended on the fimbrial interaction with heparin-binding and cell attachment domains in the fibronectin structure. The binding of P.gingivalis to mouse peritoneal macrophages via its fimbriae was inhibited by fibronectin. Fibronectin also inhibited the bacterial cell-induced expression of interleukin-1beta and KC genes in the macrophages. These results demonstrate the importance of fibronectin as a modulator of the pathogenic mechanism of P. gingivalis, a pathogen that causes adult periodontal disease.
Assuntos
Adesinas Bacterianas , Proteínas da Membrana Bacteriana Externa/metabolismo , Proteínas de Bactérias/metabolismo , Proteínas de Transporte , Quimiocinas CXC , Fibronectinas/metabolismo , Proteínas de Fímbrias , Peptídeos e Proteínas de Sinalização Intercelular , Porphyromonas gingivalis/metabolismo , Porphyromonas gingivalis/patogenicidade , Adulto , Animais , Aderência Bacteriana/fisiologia , Sítios de Ligação , Quimiocina CXCL1 , Fatores Quimiotáticos/biossíntese , Fatores Quimiotáticos/genética , Fímbrias Bacterianas/metabolismo , Expressão Gênica , Substâncias de Crescimento/biossíntese , Substâncias de Crescimento/genética , Heparina/metabolismo , Humanos , Técnicas In Vitro , Interleucina-1/biossíntese , Interleucina-1/genética , Lipopolissacarídeos/metabolismo , Macrófagos Peritoneais/imunologia , Macrófagos Peritoneais/metabolismo , Macrófagos Peritoneais/microbiologia , Camundongos , Camundongos Endogâmicos BALB C , Doenças Periodontais/etiologia , Ligação ProteicaRESUMO
White rabbits in a family, which were clinically diagnosed as moderately or severely diseased with spirochetosis, were bacteriologically and immunologically examined. The specimens from the diseased rabbits, including affected prepuces, scrotum, or skins with an occasional presence of the spirochetes, did not, however, result in growth in six conventional culture media. Serological tests, including quantitative complement fixation test, rapid plasma reagin card test, Treponema pallidum hemagglutination test, and microscopic agglutination test for leptospires using sera from diseased rabbits showed no differences when compared with those of pooled normal rabbit sera. Immunoblot analysis of the polypeptides from three human oral treponemes and three non-oral spirochetes demonstrated that antibodies against several treponemal polypeptides were detected.
Assuntos
Coelhos , Treponema/imunologia , Infecções por Treponema/veterinária , Animais , Anticorpos Antibacterianos/sangue , Antígenos de Bactérias/análise , Feminino , Immunoblotting , Masculino , Infecções por Treponema/imunologiaRESUMO
The cellular fatty acids and aldehydes of oral Eubacterium species were determined by gas chromatography-mass spectrometry. E. brachy and E. lentum contained mainly branched-chain fatty acids, whereas the others contained straight-chain acids. E. brachy, E. lentum, E. yurii ssp. yurii, E. yurii spp. margaretiae, E. limosum, E. plauti and E. aerofaciens also contained aldehydes with even carbon numbers. In addition to species-specific components, the compositional ratios of fatty acids and aldehydes characterized each individual species. The 10 species tested were divided into 5 groups by the principal component analysis. Cellular fatty acids and aldehydes would be chemical markers for interspecies differentiation of oral Eubacterium.
Assuntos
Aldeídos/análise , Eubacterium/química , Ácidos Graxos/análise , Técnicas de Tipagem Bacteriana , Eubacterium/classificação , Humanos , Boca/microbiologiaRESUMO
We tested antibacterial and antiviral activities of rat cystatin S, a cysteine proteinase inhibitor, belonging to the family 2 cystatins against 18 different bacterial species and poliovirus type 1 (Sabin). Rat cystatin S specifically inhibited the growth of a human oral anaerobic bacterium Porphyromonas gingivalis due to a bactericidal effect.
Assuntos
Cistatinas/farmacologia , Inibidores de Cisteína Proteinase/farmacologia , Porphyromonas gingivalis/efeitos dos fármacos , Sequência de Aminoácidos , Animais , Bactérias/efeitos dos fármacos , Cistatinas/química , Cistatinas/isolamento & purificação , Humanos , Testes de Sensibilidade Microbiana , Dados de Sequência Molecular , Poliovirus/efeitos dos fármacos , Sinais Direcionadores de Proteínas/química , Sinais Direcionadores de Proteínas/genética , Ratos , Cistatinas Salivares , Especificidade da Espécie , Streptococcus pyogenes/efeitos dos fármacosRESUMO
Phytoalexins, defensive compounds produced by plants against microbial infections, were purified from Sophora exigua (Leguminosae) and their growth inhibitory effects on oral cariogenic bacteria were determined in vitro. Among three isolated compounds, 5,7,2',4'-tetrahydroxy-8-lavandulylflavanone completely inhibited the growth of oral bacteria including primary cariogenic mutans streptococci, other oral streptococci, actinomycetes, and lactobacilli, at concentrations of 1.56 to 6.25 micrograms/ml.
Assuntos
Bactérias/efeitos dos fármacos , Cárie Dentária/microbiologia , Flavanonas , Plantas/química , Actinomyces/efeitos dos fármacos , Bactérias/crescimento & desenvolvimento , Fabaceae , Flavonoides/isolamento & purificação , Flavonoides/farmacologia , Lactobacillus/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Estrutura Molecular , Raízes de Plantas/química , Plantas Medicinais , Streptococcus/efeitos dos fármacos , Streptococcus mutans/efeitos dos fármacosRESUMO
The polypeptides of seven strains of human treponemes were investigated by immunoblot analysis for their binding to the human placental collagens and laminin. Of the treponemal polypeptides, eleven polypeptides, 45-kDa, 49-kDa, and 62-kDa polypeptides from T. pallidum ATCC 27087, a 48-kDa polypeptide from T. phagedenis biotype Reiter, 51-kDa and 53-kDa polypeptides from T. vincentii ATCC 35580, 30-kDa, 53-kDa and 63-kDa polypeptides from T. socranskii subsp. buccale ATCC 35534, a 52-kDa polypeptide from T. denticola ATCC 35405, and a 53-kDa polypeptide from T. denticola ATCC 33520 possessed an ability to bind to the laminin, type I, III, IV, or V collagen. An intermediate-sized human oral isolate strain G7201 did not possess any laminin- or collagen-binding polypeptides. Immunoelectron microscopy using intact treponemal cells with a single collagen-binding polypeptide and the corresponding antisera demonstrated that the 51-kDa and 53-kDa polypeptides from T. vincentii, the 53-kDa polypeptide from T. socranskii subsp. buccale ATCC 35534 and the 52-kDa polypeptide from T. denticola ATCC 35405, were outer envelope proteins.
Assuntos
Aderência Bacteriana , Proteínas de Bactérias/metabolismo , Colágeno/metabolismo , Laminina/metabolismo , Treponema/fisiologia , Eletroforese em Gel de Poliacrilamida , Humanos , Immunoblotting , Imuno-Histoquímica , Microscopia Imunoeletrônica , Peso Molecular , Treponema/metabolismo , Treponema/ultraestruturaRESUMO
The cellular fatty acid compositions of 26 strains of methicillin-resistant Staphylococcus aureus (MRSA) and 17 strains of methicillin-susceptible S. aureus (MSSA) were analyzed by gas-liquid chromatography. The fatty acid compositions of the two groups were very similar with 16 identified components. The major fatty acids were Ci14 = 0, Ci15 = 0, C18 = 0 and C20 = 0. Among these fatty acids, the percentage of the Ci15 = 0 fatty acid component of MRSA strains (11.4 +/- 3.9%) was statistically higher than that of MSSA strains (6.2 +/- 2.4%) (p < 0.001). On the other hand, the percentage of the C20 = 0 fatty acid components of MRSA strains (20.2 +/- 8.8%) was statistically lower than that of MSSA strains (30.7 +/- 10.4%) (p < 0.001). The production of beta-lactamase and beta-hemolysin in both groups' strain was also unrelated to the relative amounts of the fatty acid components. These results indicated a statistical tendency for the percentage fatty acid compositions of the MRSA strains to be quantitatively different from those of the MSSA for both the Ci15 = 0 and C20 = 0 fatty acid components. Analysis of the fatty acid compositions may have an application in the differentiation of MRSA and MSSA strains.
Assuntos
Antibacterianos/toxicidade , Ácidos Graxos/análise , Resistência a Meticilina , Meticilina/toxicidade , Staphylococcus aureus/química , Staphylococcus aureus/efeitos dos fármacos , Cromatografia Gasosa , Proteínas Hemolisinas/biossíntese , Testes de Sensibilidade Microbiana , Especificidade da Espécie , Staphylococcus aureus/fisiologia , beta-Lactamases/biossínteseRESUMO
Thirteen polypeptide antigens with molecular weights ranging from 34 kDa to 83 kDa were selected and their antigenic behaviors and distribution were examined in 12 strains of microorganisms including Treponema, Borrelia, Leptospira and Leptonema. Immunoblot analysis demonstrated that 45 kDa and 83 kDa polypeptides of Treponema socranskii subsp. buccale ATCC 35534, 53 kDa antigen of Treponema denticola ATCC 33520 and 44 kDa polypeptide of the strain G7201 were strain-specific. The 34, 62, 66 and 84 kDa polypeptide antigens were detected in all 8 treponemal strains examined. T. denticola ATCC 33520 and ATCC 35404 possessed 38 kDa, 48 kDa, 52 kDa and 72 kDa common polypeptide antigens. All 12 strains possessed the 84 kDa polypeptide antigen. Immunoelectron microscopy demonstrated that the 34 kDa and 38 kDa polypeptide antigens were located on the axial flagella and that other polypeptide antigens were located on the outer envelopes or wall-membrane complexes.
Assuntos
Antígenos de Bactérias/análise , Proteínas de Bactérias/análise , Treponema/imunologia , Animais , Anticorpos Antibacterianos , Proteínas da Membrana Bacteriana Externa/análise , Flagelos/imunologia , Humanos , Immunoblotting , Camundongos , Microscopia Imunoeletrônica , Treponema/ultraestruturaRESUMO
Two polypeptide antigens with molecular sizes of 34,000 daltons (34 kDa) and 38 kDa were separated from heated cells of a human clinical treponeme strain G7201 and Treponema denticola ATCC 35404, respectively. The rabbit polyclonal antisera against these antigens were produced and examined for their immunological reactions with the two heated antigens or intact spirochetal cells. Immunoblot analysis showed that the 34-kDa protein was also detected in T. denticola ATCC 35404 and ATCC 33520, and the 38-kDa protein was detected only in the two ATCC strains. Immunoelectron microscopy using the two rabbit antisera and protein A-gold complexes demonstrated that the 38-kDa protein antigen was present on the axial flagella of two T. denticola strains, and that the 34-kDa protein was located in the axial flagella of the G7201 cell, but neither in axial flagella nor on outer envelopes of the two ATCC strains cells, suggesting that the native 34-kDa axial flagellar protein of the G7201 strain may be different from that of T. denticola in terms of immunological reactivity.
Assuntos
Antígenos de Bactérias/imunologia , Proteínas de Bactérias/imunologia , Flagelos/imunologia , Treponema/imunologia , Reações Cruzadas , Eletroforese em Gel de Poliacrilamida , Flagelos/ultraestrutura , Humanos , Immunoblotting , Microscopia Imunoeletrônica , Treponema/ultraestruturaRESUMO
Major polypeptides from a human oral spirochete Treponema denticola ATCC 33520 were examined to demonstrate their ability to bind to human plasma fibronectin by immunoblot analysis. Of three main polypeptides separated on sodium dodecyl sulfate polyacrylamide gels 53,000-daltons (53-kDa) and 72-kDa surface antigenic proteins and a 38-kDa axial flagellar protein showed the ability to bind to fibronectin, suggesting that fibronectin on host cells can mediate cytoadherence of T. denticola by its binding to the surface proteins or the exposed 38-kDa axial flagellar protein.
Assuntos
Adesinas Bacterianas , Proteínas da Membrana Bacteriana Externa/metabolismo , Proteínas de Bactérias , Proteínas de Transporte , Treponema/metabolismo , Sequência de Aminoácidos , Antígenos de Bactérias/química , Antígenos de Bactérias/metabolismo , Aderência Bacteriana/fisiologia , Proteínas da Membrana Bacteriana Externa/química , Proteínas da Membrana Bacteriana Externa/imunologia , Sítios de Ligação , Fibronectinas/química , Fibronectinas/metabolismo , Humanos , Immunoblotting , Técnicas In Vitro , Dados de Sequência Molecular , Peso Molecular , Boca/microbiologia , Oligopeptídeos/química , Oligopeptídeos/metabolismo , Oligopeptídeos/farmacologia , Treponema/imunologia , Treponema/isolamento & purificaçãoRESUMO
In a series of investigations into the cariogenicity of Streptococcus mutans, we studied the incorporation of exogenous fatty acids with reference to glucosyltransferase secretion and membrane fatty acid changes. When cells were grown with different fatty acids, both saturated and unsaturated fatty acids were readily incorporated into the membrane lipids and were biotransformed and elongated preferentially to the longer 16- and 18-carbon-chain fatty acids. This incorporation and chain-elongation led to significant changes in fatty acids composition. By adding fatty acids to the medium, it was possible to appropriately modify the degree of unsaturation and the relative ratio between specific fatty acids in the membrane lipids of S. mutans.
Assuntos
Ácidos Graxos/farmacocinética , Streptococcus mutans/metabolismo , Ácidos Graxos/análise , Lipídeos de Membrana/análiseRESUMO
The culture fluids of various anaerobic bacteria induced the synthesis of early antigens (EA) in Epstein-Barr virus (EBV) carrying lymphoblastoid cells. The culture fluids of Corynebacterium butyricum and Fusobacterium nucleatum were the effective inducer on EA. The inducing activity was, to some extent, dependent on their n-butyric acid content, but appeared to be regulated by yet unidentified materials.
Assuntos
Corynebacterium/metabolismo , Ácidos Graxos/metabolismo , Fusobacterium/metabolismo , Herpesvirus Humano 4/fisiologia , Antígenos Virais/biossíntese , Butiratos/metabolismo , Ácido Butírico , Herpesvirus Humano 4/metabolismo , Humanos , Boca/microbiologia , Células Tumorais Cultivadas , Ativação ViralRESUMO
Profiles of metabolic alpha-keto acids were determined by a high-performance liquid chromatographic method and applied to characterization of oral black-pigmented Bacteroides. Each bacterial strain was incubated with amino acids in a chemically defined medium. After production alpha-keto acids were purified by hydrazide gel column treatment and converted to u.v.-absorbing derivatives. They were analysed by reversed-phase ion-pair chromatography. Black-pigmented Bacteroides species were differentiated into two groups according to production of aromatic alpha-keto acids. Bacteroides gingivalis, B. endodontalis and B. loescheii produced both p-hydroxyphenylpyruvic and phenylpyruvic acids. However, no such alpha-keto acids were produced by B. levii, B. intermedius and B. denticola. In addition, production profiles of several aliphatic alpha-keto acids (alpha-ketoglutaric, pyruvic, alpha-ketobutyric, alpha-ketoisovaleric, alpha-ketoisocaproic, and alpha-keto-beta-methylvaleric acids) separated each individual species in such groups. The present study offers useful chemotaxonomic information on amino acid metabolic activity of oral black-pigmented Bacteroides species.
Assuntos
Aminoácidos/metabolismo , Bacteroides/metabolismo , Cetoácidos/metabolismo , Bacteroides/classificação , Cromatografia Líquida de Alta Pressão , Humanos , Cetoácidos/análise , Cetoácidos/isolamento & purificação , Boca/microbiologia , Especificidade da EspécieRESUMO
Human oral spirochetes are prominent inhabitants of subgingival plaque in patients with periodontal disease. By immunoelectron microscopy using protein A-gold complexes and either polyclonal mouse antiserum against the 53-kDa antigen or 53-kDa-antigen-specific monoclonal antibody, a major polypeptide antigen, with a molecular weight of 53,000 (molecular size, 53 kilodaltons [kDa]), of a human oral spirochete, Treponema denticola ATCC 33520, was found to localize on the surface of the outer envelope.
Assuntos
Antígenos de Bactérias/isolamento & purificação , Antígenos de Superfície/isolamento & purificação , Treponema/imunologia , Testes de Aglutinação , Anticorpos Antibacterianos , Anticorpos Monoclonais , Especificidade de Anticorpos , Antígenos de Bactérias/imunologia , Antígenos de Superfície/imunologia , Humanos , Imuno-Histoquímica , Microscopia Eletrônica , Doenças Periodontais/microbiologia , Doenças Periodontais/patologia , Treponema/ultraestruturaRESUMO
1. Both Tween 80 and sodium fluoride significantly enhanced total extracellular glucosyltransferase activities of Streptococcus mutans. 2. Water-insoluble and water-soluble glucan formation were uniformly increased by Tween 80, whereas fluoride stimulated only water-soluble glucan formation. 3. Elevated glucan formation was due to an increase in enzymes secreted from bacterial cells. 4. Fatty acid composition and phospholipid content in bacterial membrane were changed by Tween 80, although sodium fluoride scarcely showed these changes. 5. Comparative results suggest that modulation of membrane lipids participates in mutansucrase production but not in dextransucrase production of S. mutans.