The presence of distinct bacterial species is found to be dependent on age, diet, and disease. We compared the detection rate of several oral bacterial strains in a cohort of 36 subjects including healthy volunteers, periodontal patients, and oral cancer patients. Gargling samples were obtained from these subjects from which DNA was then extracted. Specific primers for 29 bacterial species were used for PCR detection. In the oral cancer patients, Capnocytophaga ochracea, Gemella morbillorum, and Streptococcus salivarius were detected more frequently compared with the healthy volunteers and periodontitis patients. Fusobacterium nucleatum/polymorphym and Prevotella nigrescens were significantly less prevalent in oral cancer patients than the other groups. In periodontitis patients, Porphyromonas gingivalis and Treponema denticola were more frequently found compared with the healthy volunteers. In the healthy volunteer group, Peptostreptococcus anaerobius was more frequently found than the other groups. The detection rate of several oral bacterial species was thus found to differ between healthy volunteers, periodontitis patients and oral cancer patients.
Capnocytophaga ; Cohort Studies ; Diet ; DNA ; Fusobacterium ; Gemella ; Healthy Volunteers* ; Humans ; Microbiota* ; Mouth Neoplasms* ; Peptostreptococcus ; Periodontitis* ; Polymerase Chain Reaction ; Porphyromonas gingivalis ; Prevotella nigrescens ; Streptococcus ; Treponema denticola

No abstract available.
Acupuncture Therapy ; Ampicillin/pharmacology/therapeutic use ; Ankle/ultrasonography ; Anti-Bacterial Agents/pharmacology/therapeutic use ; Cellulitis/complications/diagnosis/drug therapy/*microbiology ; Diabetes Mellitus, Type 2/complications ; Gram-Negative Bacterial Infections/complications/diagnosis/drug therapy/*microbiology ; Humans ; Hypertension/complications ; Magnetic Resonance Imaging ; Male ; Microbial Sensitivity Tests ; Middle Aged ; Prevotella nigrescens/drug effects/*genetics/isolation & purification ; RNA, Ribosomal, 16S/*analysis ; Sulbactam/pharmacology/therapeutic use ; Tomography, X-Ray Computed

It has been established that berberine has strong antimicrobial effects. Little is known however regarding the antimicrobial activity of berberine against endodontic pathogenic bacteria or its cytotoxicity in human oral tissue cells. The antibacterial properties of berberine were tested against 5 strains of Enterococcus faecalis and type strains of Aggregatibacter actinomycetemcomitans, Prevotella nigrescens, Prevotella intermedia, and Tannerella forsythia, which are involved in endodontic infections. Antimicrobial activity was evaluated through minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) measurements. The viability of normal human gingival fibroblast (NHGF) cells after exposure to berberine was measured using a methyl thiazolyl tetrazolium (MTT) assay. The data showed that berberine has antimicrobial effects against A. actinomycetemcomitans with an MIC and MBC of 12.5 microg/ml and 25 microg/ml, respectively. In the cytotoxicity studies, cell viability was maintained at 66.1% following exposure to 31.3 microg/ml berberine. Overall, these findings suggest that berberine has antimicrobial activity against the tested bacteria. Nevertheless, lower concentrations in combination with other reagents will need to be tested before these in vitro results can be translated to clinical use.
Aggregatibacter ; Bacteria* ; Berberine* ; Cell Survival ; Enterococcus faecalis ; Fibroblasts ; Forsythia ; Humans ; Indicators and Reagents ; Microbial Sensitivity Tests ; Prevotella intermedia ; Prevotella nigrescens

The DNA probes Pn17 and Pn34 were evaluated for their ability to specifically detect clinical strains of P. intermedia and P. nigrescens from a Korean population by dot blot hybridization. These probes were sequenced by extension termination and their specificity was determined by Southern blot analysis. The results revealed that the Pn17 sequence (2,517 bp) partially encodes an RNA polymerase beta subunit (rpoB) and that Pn34 (1,918 bp) partially encodes both rpoB (1-169 nts) and the RNA polymerase beta subunit (rpoB'; 695-1918 nts). These probes hybridized with both HindIII- and PstI-digested genomic DNAs from the strains of P. intermedia and P. nigrescens used in this study. Interestingly, each of the hybrid bands generated from the HindIII-digested genomic DNAs of the two bacterial species could be used to distinguish between them via restriction fragment length polymorphism. These results thus indicate that Pn17 and Pn34 can simultaneously detect P. intermedia and P. nigrescens.
Blotting, Southern ; Chimera ; DNA ; DNA Probes ; DNA-Directed RNA Polymerases ; Polymorphism, Restriction Fragment Length ; Prevotella ; Prevotella intermedia ; Prevotella nigrescens ; Sensitivity and Specificity

PURPOSE: Interleukin-8 (IL-8) is an important mediator of immune and inflammatory reactions and is produced by a variety of different cell types. This study was undertaken to investigate the effects of lipopolysaccharides (LPSs) from Prevotella intermedia and Prevotella nigrescens, the major causes of inflammatory periodontal disease, on the production of IL-8 and the expression of IL-8 mRNA in differentiated THP-1 cells, a human monocytic cell line. METHODS:LPSs from P. intermedia ATCC 25611 and P. nigrescens ATCC 33563 were prepared by the standard hot phenol-water method. THP-1 cells were incubated in the medium supplemented with phorbol myristate acetate to induce differentiation into macrophage-like cells. RESULTS: We found that LPS preparations from P. intermedia and P. nigrescens can induce IL-8 mRNA expression and stimulate the release of IL-8 in differentiated THP-1 cells without additional stimuli. CONCLUSIONS: There are no previous reports of the ability of P. intermedia and P. nigrescens LPS to stimulate the release of IL-8, and the present study clearly shows, for the first time, that LPSs from P. intermedia and P. nigrescens fully induced IL-8 mRNA expression and IL-8 production in differentiated human monocytic cell line THP-1. The ability of P. intermedia and P. nigrescens LPS to promote the production of IL-8 may be important in the pathogenesis of inflammatory periodontal disease.
Cell Line ; Humans ; Interleukin-8 ; Lipopolysaccharides ; Macrophages ; Periodontal Diseases ; Phorbols ; Prevotella ; Prevotella intermedia ; Prevotella nigrescens ; RNA, Messenger ; Tetradecanoylphorbol Acetate

PURPOSE: Specific bacteria are believed to play an important role in chronic periodontitis. Although extensive microbial analyses have been performed from subgingival plaque samples of periodontitis patients, systemic analysis of subingival microbiota has not been carried out in a Korean population so far. The purpose of this study was to investigate the prevalence of 29 putative periodontal pathogens in Korean chronic periodontitis patients and evaluate which pathogens are more associated with Korean chronic periodontitis. MATERIAL AND METHODS: A total of 86 subgingival plaque samples were taken from 15 chronic periodontits(CP) patients and 13 periodontally healthy subjects in Korea. CP samples were obtained from the deepest periodontal pocket (>3 mm probing depth[PD]) and the most shallow periodontal probing site (< or =3 mm PD) in anterior tooth and posterior tooth, respectively, of each patient. Samples in healthy subjects were obtained from 1 anterior tooth and 1 posterior tooth. Polymerase chain reaction (PCR) of 16S ribosomal DNA (rDNA) of subgingival plaque bacteria was performed. Detection frequencies(% prevalence) of 29 putative periodontal pathogens were investigated as bacterium-positive sites/total sites RESULTS: With the exception of Olsenella profuse and Prevotella nigrescens, the sites of diseased patients generally showed higher prevalence than the healthy sites of healthy subjects for all bacteria analyzed. Tanerella forsythensis (B.forsythus), Campylobacter rectus, Filifactor alocis, Fusobacterium nucleatum, Porphyromonas endodontalis and Porphyromonas gingivalis were detected in more than 80% of sites with deep probing depths in CP patients. In comparison between the sites (deep or shallow PD) of CP patients and the healthy sites of healthy subjects, there was statistically significant difference(P <0.05) of prevalence in T.forsythensis (B.forsythus), C.rectus, Dialister invisus, F.alocis, P.gingivalis and Treponema denticola. CONCLUSION: Our results demonstrate that the four putative periodontal pathogens, T.forsythensis (B.forsythus), C.rectus, P.gingivalis and F.alocis are closely related with CP patients in the Korean population.
Bacteria ; Campylobacter rectus ; Chronic Periodontitis ; DNA, Ribosomal ; Fusobacterium nucleatum ; Humans ; Korea ; Metagenome ; Periodontal Pocket ; Periodontitis ; Polymerase Chain Reaction ; Porphyromonas endodontalis ; Porphyromonas gingivalis ; Prevalence ; Prevotella nigrescens ; Tooth ; Treponema

The results of this study confirm that the availability of hemin influences the expression of selected membrane proteins of Porphyromonas gingivalis, Prevotella intermedia, and Prevotella nigrescens. A 30 kDa (heated 24 kDa) hemin-binding protein whose expression is hemin regulated was identified and purified in P. gingivalis. A strong hemin-binding function was found by LDS-PAGE and TMBZ staining when P. gingivalis cells were grown under hemin-limited conditions. A 50 kDa cell envelope associated protein, whose expression is hemin regulated, is considered to be a putative hemin binding protein from P. intermedia and P. nigrescens, respectively. N-terminal amino acid sequence analysis of CNBr-digested 24 kDa hemin binding protein from P. gingivalis revealed that this protein belongs to a new, so far undescribed hemin-binding class of proteins. N-terminal amino acid sequence of a 50 kDa putative hemin binding protein from P. intermedia was identical with Enolase from Streptococcus intermedia. Work is in progress to further characterize the molecular structure of these proteins.
Amino Acid Sequence ; Carrier Proteins ; Hemin ; Membrane Proteins ; Molecular Structure ; Phosphopyruvate Hydratase ; Porphyromonas gingivalis* ; Porphyromonas* ; Prevotella intermedia* ; Prevotella nigrescens* ; Prevotella* ; Sequence Analysis, Protein ; Streptococcus

The aim of this study was to determine the minimal inhibitory concentration(MIC) of cefi- xime, which is a 3rd generation of cefalosporin, against 6 species of putative periodon- topathogens; Fusobacterium nucleatum, Actinobacillus actinomycetemcomitans, Prevotella inter- media, Prevotella nigrescens, Tannerella forsythia and Porphyromonas gingivalis. The efficacy of cefixime was examined by comparing it with that of several antibiotics(amoxicillin, Aug- mentin(R) ciprofloxacin, metronidazole, and tetracycline), which were used as the control. The MIC was measured using a microdilution method. The MIC of cefixime against the putative periodotopathogens, as a single use regimen, was relatively lower than that of the other antibiotics. The MIC of cefixime/metronidazole against P. intermedia ChDC KB14, P. nigres- cens ChDC KB50, F. nucleatum ChDC PV-F37, F. nucleatum ChDC F130, and F. nucleatum ChDC F175, as a simultaneous regimen, was lower than that of the other antibiotics. The concentration of cefixime in the crevicular fluid of volunteers who received 250mg every 12 hours for 3 days was 9microgram/ml after 9 hours. In conclusion, cefixime showed good anti- microbial activity in a single treatment or as a combined therapy with amoxicillin, Aug- mentin(R) or metronidazole against 6 periodontopathogens.
Aggregatibacter actinomycetemcomitans ; Amoxicillin ; Anti-Bacterial Agents ; Cefixime* ; Ciprofloxacin ; Forsythia ; Fusobacterium nucleatum ; Metronidazole ; Porphyromonas gingivalis ; Prevotella ; Prevotella nigrescens ; Volunteers

The purpose of this study was to monitor the secretion of matrix metalloproteinase (MMP) and tissue inhibitor of metalloproteinase (TIMP) by human periodontal ligament (PDL) fibroblasts stimulated with Prevotella nigrescens lipopolysaccharide (LPS), and to examine the effect of calcium hydroxide treatment on P. nigrescens LPS. LPS was extracted and purified from anaerobically cultured P. nigrescens. PDL fibroblasts were stimulated by the LPS (0, 0.1, 1, 10 ug/ml) or LPS (10 ug/ml) pretreated with 12.5 mg/ml of Ca(OH)2 for 3 days, for various periods of time (12, 24, 48 h). Immunoprecipitation were performed for protein level analysis of MMP-1, MMP-2 and TIMP-1. Total RNA was isolated and real-time quantitative polymerase chain reaction (PCR) was performed for quantification of MMP-1 mRNA. According to this study, the results were as follows: 1. The production of MMP-1 by stimulation with P. nigrescens LPS increased in time-dependent manner, and showed maximum value at 48 h in both protein and mRNA level. But there was no dose-dependent increase. 2. MMP-2 production time-dependently increased when stimulated with 1 and 10 ug/ml LPS, but there was no dose-dependent increase. 3. TIMP-1 production increased to 24 h, but decreased at 48 h. It increased when stimulated with 0.1 and 1 ug/ml LPS, but suppressed at 10 ug/ml. 4. P. nigrescens LPS pretreated with Ca(OH)2 markedly downregulated MMP-1 gene expression.
Calcium Hydroxide ; Fibroblasts* ; Gene Expression ; Humans ; Immunoprecipitation ; Periodontal Ligament* ; Polymerase Chain Reaction ; Prevotella nigrescens* ; Prevotella* ; RNA ; RNA, Messenger ; Tissue Inhibitor of Metalloproteinase-1

Hemolytic property is a specific feature of bacteria to obtain iron which is essential for its survival in host tissues. Therefore, it is thought to be one of several factors of virulence. The purpose of this study was to investigate the hemolytic properties of Prevotella nigrescens isolated from the teeth diagnosed as pulp necrosis and apical periodontitis under the presence of hemolysin inhibitors such as NaN3 and dithiothreitol, heat, various pH and cultural conditions. The results were as follows; 1. Clinically isolated P. nigrescens strains and standard P. nigrscens ATCC 33563 showed hemolytic activity. 2. P. nigrescens showed higher hemolytic activity against human erythrocytes than sheep or horse erythrocytes. 3. NaN3 and dithiothreitol (DTT) reduced the hemolytic activity of P. nigrescens in a dose dependent manner (p < 0.05). 4. Optimal pH for the maximum hemolytic activity of P. nigrescens was 4.0 and the hemolysin was stable under the 50degrees C, but the hemolytic activity was significantly decreased at 95degrees C. 5. P. nigrescens cultured in 10% CO2 condition showed higher hemolytic activity than the bacteria cultured in the anaerobic condition.
Bacteria ; Dental Pulp Necrosis ; Dithiothreitol ; Erythrocytes ; Horses ; Hot Temperature ; Humans ; Hydrogen-Ion Concentration ; Iron ; Periapical Periodontitis ; Prevotella nigrescens* ; Prevotella* ; Sheep ; Sodium Azide ; Tooth ; Virulence