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The Msp protein complex and the serine protease dentilisin are the best‐characterized virulence factors in Treponema denticola, the major etiological agent of chronic periodontitis. In addition to these outer sheath factors, the cysteine protease dentipain contributes to pathogenicity, but its secretion, processing, cellular localization, and role in T. denticola virulence are not fully understood. In this study, we found that full‐sized dentipain (74‐kDa) and the 52‐kDa truncated form of the enzyme are located, respectively, in the outer sheath derived from T. denticola dentilisin‐ and the Msp‐deficient mutants. Furthermore, dentipain was barely detected in the wild‐type strain. These results suggest that dentilisin and Msp, the major outer sheath proteins, are involved in the secretion and maturation of dentipain. Inactivation of the dentipain gene slowed the growth of T. denticola, and the effect was more profound in serum‐free medium than in serum‐containing medium. Several genes, including those encoding transporters and methyl‐accepting chemotaxis proteins, were differentially expressed in the dentipain‐deficient mutant. Furthermore, the mutant strain was more hydrophobic than the wild‐type strain. Finally, the mutant showed less autoaggregation activity and adhesion to IgG in a serum‐free medium than the wild‐type strain. These findings suggest that dentipain contributes to the virulence of T. denticola by facilitating adhesion and acquisition of nutrients essential for colonization and proliferation in the gingival crevice under serum‐rich conditions. [ABSTRACT FROM AUTHOR]

Treponema denticola, a helically shaped motile microorganism, is a major pathogen of chronic periodontitis. Major surface protein (Msp) and dentilisin are virulence factors of T. denticola that are located on the outer sheath. The motility of T. denticola is deeply involved in colonization on and invasion into the host tissue. The outer sheath is located at the interface between the environment and T. denticola, and its components may also contribute to its motility via interaction with the materials outside the cells. The study aimed to clarify whether Msp or dentilisin contributes to the motility of T. denticola on solid surfaces, termed crawling, by investigating their effects using Msp‐deficient and dentilisin‐deficient T. denticola strains. Motility was analyzed by measuring the colony size in agar plates and velocity was analyzed using dark‐field microscopy. The colony area of the mutant strains was smaller than that of the wild‐type strain. The crawling velocity of the mutant strains was lower than that of the wild‐type strain, with the lowest velocity observed in the dentilisin‐deficient strain. Additionally, the ratio of the crawling distance by one revolution to the protoplasmic cylinder pitch (an indicator of the crawling efficiency) in the dentilisin mutant was significantly lower than that in the wild type strain and the Msp mutant. Together, these results indicate that dentilisin facilitates the crawling‐dependent surface spreading of T. denticola. [ABSTRACT FROM AUTHOR]

Background/objectives: Cytotoxic T lymphocyte‐associated antigen 4 (CTLA‐4) is a critical immunoregulatory molecule expressed on T cells. CTLA‐4 also binds to the surfaces of monocytes and macrophages, precursors of osteoclasts. Research on rheumatoid arthritis demonstrated that CTLA‐4 suppresses inflammation and bone resorption. However, its effects on alveolar bone have yet to be understood. The purpose of this study was to investigate the role and potential mechanism of CTLA‐4 in bone resorption in periodontitis. Materials and methods: In vivo, the effects of systemic administration of CTLA‐4 immunoglobulin fusion protein (CTLA‐4‐Ig) on alveolar bone resorption were investigated using a periodontitis mouse model. A total of 20 C57BL/6J mice were randomly assigned to two groups according to the administration modes. Periodontitis was induced by placing a ligature around the left maxillary second molar. The contralateral tooth was left un‐ligated. In the CTLA‐4‐Ig (+) group, CTLA‐4‐Ig was administered by intraperitoneal injection at 1 and 3 days after ligature placement. Animals in the CTLA‐4‐Ig (−) group were given only phosphate‐buffered saline each time. At 5 days after ligature placement, bone resorption was assessed by micro‐computed tomography and histological examination, and the prevalence of osteoclast‐like cells was assessed by tartrate‐resistant acid phosphatase (TRAP) staining. In vitro, the effects of CTLA‐4‐Ig on osteoclasts were evaluated. Viability of RAW 264.7 cells treated with receptor activator of nuclear factor‐κB ligand (RANKL) and CTLA‐4‐Ig was tested by WST‐1 assay. Osteoclast‐like cells were enumerated by TRAP staining, and osteoclast activity was evaluated by resorption pit assay. Gene expression levels of osteoclast differentiation markers (macrophage‐colony stimulating factor receptor, carbonic anhydrase II, cathepsin K, and Trap) and protein phosphatase 2A (PP2A), a major serine‐threonine phosphatase, were assessed by quantitative real‐time polymerase chain reaction. The effect of CTLA‐4‐Ig on the nuclear factor‐κB (NF‐κB) activation was assessed by enzyme‐linked immunosorbent assay. Results: In vivo, ligature‐induced bone resorption and the numbers of osteoclast‐like cells were significantly decreased by the administration of CTLA‐4‐Ig. In vitro, treatment with RANKL and CTLA‐4‐Ig had no significant effect on cell viability. CTLA‐4‐Ig significantly reduced the prevalence and activation of osteoclast‐like cells and decreased the expressions of osteoclast differentiation markers, compared with the RANKL‐treated control. CTLA‐4‐Ig significantly suppressed RANKL‐induced phosphorylation of NF‐κB p65 but increased PP2A expression. Conclusion: These results suggest that CTLA‐4‐Ig abrogates bone resorption in induced periodontitis, possibly via inhibition of osteoclast differentiation and activation. The regulation of the NF‐κB pathway and PP2A expression may be one mechanism by which CTLA‐4‐Ig suppresses osteoclast behavior. [ABSTRACT FROM AUTHOR]

Background and Objective: Dysbiosis, a loss of balance in the microbiota, is a potential factor of peri‐implantitis. However, compositional change of the peri‐implant microbiota soon after implant uncovering is still unknown. In this study, bacterial composition in the peri‐implant sulcus was examined to understand the establishment of bacterial composition within the peri‐implant microbiota during the earliest weeks after implant uncovering. Methods: Microbiota samples were collected at weeks 1, 2, 4, and 6 after stage‐two surgery. Bacterial DNA was isolated from the samples, and a 16S rRNA gene library was constructed. Sequence reads were obtained using a high‐throughput sequencing platform and were taxonomically assigned at the phylum and genus levels. Results: Alpha diversity indices, which did not include taxonomic information, were at similar levels throughout the four time points. At 1 and 2 weeks, the bacterial composition was similar among patients with the predominance of Firmicutes and Proteobacteria. However, the composition was diverse at 4 and 6 weeks and significantly dissimilar to the composition at 1 week. Conclusions: At 1 week, the peri‐implant microbiota was already formed with alpha diversity as high as that at the later time points. However, the bacterial composition was not highly dissimilar among patients at 1 week. The composition changed over the passage of several weeks and was specific for each patient. [ABSTRACT FROM AUTHOR]

ObjectiveTreponema denticola is involved in 'chronic' periodontitis pathogenesis. The mechanism underlying the regulation of the expression of its virulence factors, such as major surface protein (Msp) and prolyl-phenylalanine specific protease (dentilisin) is yet to be clarified. We determined the gene expression profiles of Msp- and dentilisin-deficient mutants of T. denticola to identify the regulation network of gene expression concomitant with the inactivation of these virulence genes. Methods Gene expression profiles of T. denticola ATCC 35405 (wild type), dentilisin-deficient mutant K1, and msp-deficient mutant DMSP3 were determined using DNA microarray analysis and quantitative real-time reverse transcription PCR (qRT-PCR). Msp and dentilisin protein levels were determined by immunoblotting and proteolytic activity assays. Results In addition to several differentially expressed genes, dentilisin expression was reduced in DMSP3; msp expression was significantly reduced in K1 (p < 0.05), both at the gene and protein levels. To identify the regulatory system involved, the expression levels of the potential regulators whose expression showed changes in the mutants were evaluated using qRT-PCR. Transcriptional regulators TDE_0127 and TDE_0814 were upregulated in K1, and the potential repressor, TDE_0344, was elevated in DMSP3. Conclusions Dentilisin and Msp expression were interrelated, and gene expression regulators, such as TDE_0127, may be involved in their regulation. [ABSTRACT FROM AUTHOR]

Purpose: The presence of periodontopathic bacteria is a risk factor for peri-implantitis. The present study examined colonization by periodontopathic bacteria and their transmission from periodontal pockets to osseointegrated implant sulcus. Materials and Methods: Plaque samples were collected from 105 sites in the 15 patients who participated in the study. Colonization by these bacteria was examined by polymerase chain reaction (PCR) and culture. The transmission of periodontopathic bacteria from periodontal sites of natural teeth to the implant sulcus was analyzed by pulsed field gel electrophoresis (PFGE). Results: The PCR detection rates of Porphyromonas gingivalis, Prevotella intermedia, Actinobacillus actinomycetemcomitans, Bacteroides forsythus, and Treponema denticola were 80.0%, 53.3%, 46.7%, 60.0% and 40.0%, respectively. Colonizations by P gingivalis and A actinomycetemcomitans were statistically correlated with periodontal pockets and implant sulcus regions (P < .01). The PFGE patterns of the P gingivalis strains isolated from each patient were identical, but differed from those from other patients. The PFGE patterns of P intermedia strains were identical in 2 out of 3 patients. Discussion: These analyses indicated that there appeared to be transmission of P gingivalis and P intermedia from the periodontal pocket to the peri-implant region. Conclusion: Elimination of these periodontal pathogens from the patient's oral cavity before administering dental implant treatment may inhibit colonization by these pathogens and reduce the risk of peri-implantitis. [ABSTRACT FROM AUTHOR]

Objective: Growing evidence indicates an association between periodontitis and delivery outcome; however, the mechanism is unclear. This study aimed to investigate the influence of Porphyromonas gingivalis (Pg) infection on delivery outcome in mice.Materials and Methods: Bacteremia was induced in pregnant Slc:ICR mice (8 weeks old) by intravenous injection of Pg. Mice were randomly divided into a control group (CO), and those receiving Pg injection at gestational day 1 (GD1), gestational day 15 (GD15) or every day (ED). Delivery outcome, Pg infection, and gene expression in the placenta and umbilical cord were evaluated.Results: Birth weight was lower in the ED and GD15 groups than in the CO group. A remarkable increase in anti-Pg IgG antibody was observed in the ED and GD1 groups, although Pg was not detected in the placenta or umbilical cord. mRNA expression of Tnfα and Il6 in the placenta, and Hif1α in the umbilical cord, was significantly increased in the ED group. Microarray analysis of the umbilical cord revealed increased expression of several genes including Orm1, Mgl2, Rps6ka3 and Trim15 in the ED group.Conclusions: Pg infection during the third trimester caused low birth weight and inflammation in the placenta and umbilical cord. [ABSTRACT FROM AUTHOR]

Interaction between two periodontal pathogens, Porphyromonas gingivalis and Treponema denticola, contributes to plaque biofilm formation. Porphyromonas gingivalis forms aggregates with T. denticola through its adhesion/hemagglutinin domain (Hgp44). In this study, we investigated the specific domain of P. gingivalis Hgp44 responsible for adhesion to T. denticola using expression vectors harboring P. gingivalis Hgp44 DNA sequences encoding amino acid residues 1–419. Six plasmids harboring fragments in this region were generated by PCR amplification and self-ligation, and recombinant proteins r-Hgp44 (residues 1–419), r-Hgp441 (residues 1–124), r-Hgp442 (1–199), r-Hgp443 (1–316), r-Hgp444 (199–419), r-Hgp445 (124–198) and r-Hgp446 (199–316) were produced, as confirmed by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and immunoblotting. r-Hgp44, r-Hgp443 and r-Hgp446 showed greater adhesion to T. denticola sonicates than the control, as determined by enzyme-linked immunosorbent assay. r-Hgp446 reduced the coaggregation of P. gingivalis and T. denticola. Scanning electron and confocal laser scanning microscopy analyses revealed that r-Hgp446 reduced dual-species biofilm formation. Our results indicate that residues 199–316 of P. gingivalis Hgp44 are mainly responsible for adhesion to T. denticola; inhibiting this domain could potentially disrupt periodontopathic biofilm formation and maturation. Porphyromonas gingivalis Hgp44 residues 199–316 are responsible for adhesion to Treponema denticola; inhibition of this domain may disrupt periodontopathic biofilm formation and maturation. [ABSTRACT FROM AUTHOR]

Porphyromonas gingivalis is a bacterium frequently isolated from chronic periodontal lesions and is involved in the development of chronic periodontitis. To colonize the gingival crevice, P. gingivalis has to adapt to environmental stresses. Microbial gene expression is regulated by transcription factors such as those in two-component systems and extracytoplasmic function (ECF) sigma factors. ECF sigma factors are involved in the regulation of environmental stress response genes; however, the roles of individual ECF sigma factors are largely unknown. The purpose of this study was to investigate the functions, including autoaggregation, hemagglutination, gingipain activity, susceptibility to antimicrobial agents, and surface structure formation, of P. gingivalis ECF sigma factors encoded by SigP (PGN_0274), SigCH (PGN_0319), PGN_0450, PGN_0970, and SigH (PGN_1740). Various physiological aspects of the sigP mutant were affected; autoaggregation was significantly decreased at 60 min (p < 0.001), hemagglutination activity was markedly reduced, and enzymatic activities of Kgp and Rgps were significantly decreased (p < 0.001). The other mutants also showed approximately 50% reduction in Rgps activity. Kgp activity was significantly reduced in the sigH mutant (p < 0.001). No significant differences in susceptibilities to tetracycline and ofloxacin were observed in the mutants compared to those of the wild-type strain. However, the sigP mutant displayed an increased susceptibility to ampicillin, whereas the PGN_0450 and sigH mutants showed reduced susceptibility. Transmission electron microscopy images revealed increased levels of outer membrane vesicles formed at the cell surfaces of the sigP mutant. These results indicate that SigP is important for bacterial surface-associated activities, including gingipain activity, autoaggregation, hemagglutination, vesicle formation, and antimicrobial susceptibility. [ABSTRACT FROM AUTHOR]

Aim The purpose of the present study was to evaluate the application possibility of water containing organic acids ( WOA), made by some organic acids used as food additives, for chemical denture cleaning for older adults by microbial investigation. Methods Using an in vitro biofilm study, we determined the effects of WOA on Streptococcus sanguinis, S. pneumoniae and Candida albicans attached to heat-cured acrylic resins. Specimens were divided into three groups as follows: control group ( TW), commercial denture cleaner group ( DC) and WOA group ( WOA). Specimens were treated with each for 5 min, 30 min or 8 h, and the numbers of attached microbes were determined by counting colony-forming units or adenosine triphosphate analysis. Using an in vivo biofilm study, we studied the effects of these same solutions on 60 complete dentures. The dentures were divided randomly and blindness into three groups as described above, and treated for 10 min. The numbers of microbes attached to dentures before and after treatment were determined by counting colony-forming units. Results For the in vitro biofilm study, there were significant differences in the numbers of microbes between WOA and TW, although there were no significant differences between WOA and DC except for C. albicans. For the in vivo biofilm study, there were significant differences between WOA, DC and TW, although there was no significant difference between WOA and DC. Conclusion We conclude that water containing organic acids exerts antimicrobial effects as strong as commercial denture cleaner, and it has an application possibility of use for safe chemical denture cleaning for older adults. Geriatr Gerontol Int 2015; ●●: ●●-●●. [ABSTRACT FROM AUTHOR]

Capnocytophaga ochracea is present in the dental plaque biofilm of patients with periodontitis. Biofilm cells change their phenotype through quorum sensing in response to fluctuations in cell-population density. Quorum sensing is mediated by auto-inducers (AIs). AI-2 is involved in intercellular signaling, and production of its distant precursor is catalyzed by LuxS, an enzyme involved in the activated methyl cycle. Our aim was to clarify the role of LuxS in biofilm formation by C. ochracea. Two luxS-deficient mutants, TmAI2 and LKT7, were constructed from C. ochracea ATCC 27872 by homologous recombination. The mutants produced significantly less AI-2 than the wild type. The growth rates of these mutants were similar to that of the wild-type in both undiluted Tryptic soy broth and 0.5 × Tryptic soy broth. However, according to crystal violet staining, they produced significantly less biofilm than the wild type. Confocal laser scanning microscopy and scanning electron microscopy showed that the biofilm of the TmAI2 strain had a rougher structure than that of the wild type. Complementation of TmAI-2 with extrinsic AI-2 from the culture supernatant of wild-type strain did not restore biofilm formation by the TmAI2 strain, but complementation of LKT7 strain with luxS partially restored biofilm formation. These results indicate that LuxS is involved in biofilm formation by C. ochracea, and that the attenuation of biofilm formation by the mutants is likely caused by a defect in the activated methyl cycle rather than by a loss of AI-2. [ABSTRACT FROM AUTHOR]

Background: A number of studies have revealed a link between chronic periodontitis and cardiovascular disease in obese patients. However, there is little information about the influence of periodontitis-associated bacteria, Porphyromonas gingivalis (Pg), on pathogenesis of atherosclerosis in obesity. Methods: In vivo experiment: C57BL/6J mice were fed with a high-fat diet (HFD) or normal chow diet (CD), as a control. Pg was infected from the pulp chamber. At 6 weeks post-infection, histological and immunohistochemical analysis of aortal tissues was performed. In vitro experiment: hTERT-immortalized human umbilical vein endothelial cells (HuhT1) were used to assess the effect of Pg/Pg-LPS on free fatty acid (FFA) induced endothelial cells apoptosis and regulation of cytokine gene expression. Results: Weaker staining of CD31 and increased numbers of TUNEL positive cells in aortal tissue of HFD mice indicated endothelial injury. Pg infection exacerbated the endothelial injury. Immunohistochemically, Pg was detected deep in the smooth muscle of the aorta, and the number of Pg cells in the aortal wall was higher in HFD mice than in CD mice. Moreover, in vitro, FFA treatment induced apoptosis in HuhT1 cells and exposure to Pg-LPS increased this effect. In addition, Pg and Pg-LPS both attenuated cytokine production in HuhT1 cells stimulated by palmitate. Conclusions: Dental infection of Pg may contribute to pathogenesis of atherosclerosis by accelerating FFA-induced endothelial injury. [ABSTRACT FROM AUTHOR]

Csa2 is a member of both the Candida albicans Rbt5 protein family and the Common in Fungal Extracellular Membranes ( CFEM) protein superfamily. CFEM proteins are characterized by an internal domain containing eight equally spaced cysteine residues. Csa2 is involved in iron uptake from hemoglobin and heme proteins; however, its precise role is unclear. Here, we provide quantitative evidence of the involvement of Csa2 in the utilization of iron from human hemoglobin during C. albicans hyphal growth. The ability of the hyphal form of the wild-type (wt), a homozygote csa2Δ mutant, and a complemented strain of C. albicans to utilize hemoglobin as an iron source under iron-restricted conditions was examined through growth studies and a crystal violet-staining assay. Hemoglobin-binding activity was assessed indirectly using a hemoglobin-sensitized tube method. Although hyphal growth of the wt and csa2Δ/Δ:: CSA2 strains was completely recovered when a high concentration of human hemoglobin was added to the iron-restricted culture medium, the recovery of the csa2Δ/Δ mutant was significantly diminished. Furthermore, hemoglobin binding was impaired in the csa2Δ/Δ mutant compared with the wt and csa2Δ/Δ:: CSA2 strains, revealing that Csa2 is involved in the utilization of hemoglobin as an iron source by the hyphal form of C. albicans. [ABSTRACT FROM AUTHOR]

Vinculin, a 116-kDa membrane cytoskeletal protein, is an important molecule for cell adhesion; however, little is known about its other cellular functions. Here, we demonstrated that vinculin binds to Rab5 and is required for Staphylococcus aureus (S. aureus) uptake in cells. Viunculin directly bound to Rab5 and enhanced the activation of S. aureus uptake. Over-expression of active vinculin mutants enhanced S. aureus uptake, whereas over-expression of an inactive vinculin mutant decreased S. aureus uptake. Vinculin bound to Rab5 at the N-terminal region (1-258) of vinculin. Vinculin and Rab5 were involved in the S. aureus-induced phosphorylation of MAP kinases (p38, Erk, and JNK) and IL-6 expression. Finally, vinculin and Rab5 knockdown reduced infection of S. aureus, phosphorylation of MAPKs and IL-6 expression in murine lungs. Our results suggest that vinculin binds to Rab5 and that these two molecules cooperatively enhance bacterial infection and the inflammatory response. [ABSTRACT FROM AUTHOR]

ABSTRACT Purpose: Prevention of peri-implantitis is essential for the success of implant rehabilitation. Infection by periodontopathic bacteria is a major cause of peri-implantitis. The aim of the present study was to identify the source of peri-implant colonization by periodontopathic bacteria. Materials and Methods: Twenty-one patients with implants were enrolled in the study. Subgingival plaque samples from the adjacent, occluding, and contralateral natural teeth were collected prior to second-stage surgery. Samples from implant sulci were then obtained 2 weeks later. Detection of periodontopathic bacteria was performed by the polymerase chain reaction. Results: The detection rates for Aggregatibacter actinomycetemcomitans, Prevotella intermedia, Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia, and Fusobacterium nucleatum in all subgingival samples from natural teeth were similar to that in the peri-implant sulci. Multiple logistic regression analysis revealed an association between the detection of A. actinomycetemcomitans, Prevotella intermedia, Porphyromonas gingivalis, Treponema denticola, and F. nucleatum in the gingival crevices of adjacent teeth and that of the peri-implant sulcus, but no association for Tannerella forsythia. Conclusions: The present findings suggest that colonization by A. actinomycetemcomitans, Prevotella intermedia, Porphyromonas gingivalis, Treponema denticola, and F. nucleatum at the implant sulcus was affected by these microorganisms in the gingival crevice of adjacent teeth rather than those on occluding and contralateral teeth. [ABSTRACT FROM AUTHOR]

doi: 10.1111/j.1741-2358.2011.00506.x Colonisation of the oral cavity by periodontopathic bacteria in complete denture wearers Objective: The purpose of this study was to investigate colonisation by periodontopathic bacteria and the sites of colonisation in elderly upper and lower complete denture wearers. We also investigated the relationship between level of oral hygiene and colonisation by periodontopathic bacteria. Materials and methods: Forty edentulous and 37 dentate volunteers participated in this study. Samples were collected from whole saliva, and levels of Aggregatibacter actinomycetemcomitans, Prevotella intermedia, Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia and Fusobacterium nucleatum were determined by PCR Invader technology. Detection of these species on oral mucosal and denture surfaces was performed by PCR. Fisher's exact test was used for the statistical analysis. Cluster analysis was employed to investigate trends in the periodontopathic bacteria flora in each sampling area. Results: Detection rates of periodontopathic bacteria in whole saliva were lower under edentulous conditions than under dentulous conditions, except for A. actinomycetemcomitans and F. nucleatum ( p < 0.01). Detection rate of F. nucleatum was the highest in all areas. A positive correlation was observed between DNA quantification of P. gingivalis and number of Candida species in saliva. Cluster analysis of the test species identified two clusters. Tongue-coating status was associated with the detection rate of all periodontopathic bacteria investigated, and denture plaque status was associated with the detection rate of T. denticola and F. nucleatum. Conclusion: Results indicate the presence of periodontopathic bacteria under edentulous conditions and that the status of oral hygiene of the mucosal or denture surfaces affects colonisation by T. denticola and F. nucleatum. [ABSTRACT FROM AUTHOR]

Background: Smoking is a risk factor for periodontitis. To clarify the contribution of smoking to periodontitis, it is essential to assess the relationship between smoking and the subgingival microflora. The aim of this study was to gain an insight into the influence of smoking on the microflora of Japanese patients with periodontitis. Methods: Sixty-seven Japanese patients with chronic periodontitis (19 to 83 years old, 23 women and 44 men) were enrolled in the present study. They consisted of 30 smokers and 37 non-smokers. Periodontal parameters including probing pocket depth (PPD) and bleeding on probing (BOP) and oral hygiene status were recorded. Detection of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia, Fusobacterium nucleatum/periodonticum, Treponema denticola and Campylobacter rectus in subgingival plaque samples was performed by polymerase chain reaction. Association between the detection of periodontopathic bacteria and smoking status was analyzed by multiple logistic regression analysis and chi-square test. Results: A statistically significant association was found between having a PPD ≥ 4 mm and detection of T. denticola, P. intermedia, T. forsythia, or C. rectus, with odds ratios ranging from 2.17 to 3.54. A significant association was noted between BOP and the detection of C. rectus or P. intermedia, and smoking, with odds ratios ranging from 1.99 to 5.62. Prevalence of C. rectus was higher in smokers than non-smokers, whereas that of A. actinomycetemcomitans was lower in smokers. Conclusions: Within limits, the analysis of the subgingival microbial flora in smokers and non-smokers with chronic periodontitis suggests a relevant association between smoking and colonization by the specific periodontal pathogens including C. rectus. [ABSTRACT FROM AUTHOR]

Porphyromonas gingivalis and Treponema denticola are major pathogens of periodontal disease. Coaggregation between microorganisms plays a key role in the colonization of the gingival crevice and the organization of periodontopathic biofilms. We investigated the involvement of surface ligands of P. gingivalis in coaggregation. Two triple mutants of P. gingivalis lacking Arg-gingipain A (RgpA), Lys-gingipain (Kgp) and Hemagglutinin A (HagA) or RgpA, Arg-gingipain B (RgpB) and Kgp showed significantly decreased coaggregation with T. denticola, whereas coaggregation with a major fimbriae (FimA)-deficient mutant was the same as that with the P. gingivalis wild-type parent strain. rgpA, kgp and hagA code for proteins that contain 44 kDa Hgp44 adhesin domains. The coaggregation activity of an rgpA kgp mutant was significantly higher than that of the rgpA kgp hagA mutant. Furthermore, anti-Hgp44 immunoglobulin G reduced coaggregation between P. gingivalis wild type and T. denticola. Treponema denticola sonicates adhered to recombinant Rgp domains. Coaggregation following co-culture of the rgpA kgp hagA mutant expressing the RgpB protease with the rgpA rgpB kgp mutant expressing the unprocessed HagA protein was enhanced compared with that of each triple mutant with T. denticola. These results indicate that the processed P. gingivalis surface Hgp44 domains are key adhesion factors for coaggregation with T. denticola. [ABSTRACT FROM AUTHOR]

The article focuses on the virulence factors of Treponema denticola including major outer sheath protein (Msp) and dentilisin which were analyzed using molecular genetic techniques. It says that development of animal models improves the apprehension of pathogenicity of T. denticola which allows detailed analysis and interaction with other periodontopathic bacteria in developing periodontitis. It adds that virulence factors of T. denticola are associated with chronic periodontitis development.

Treponema denticola is a major pathogen of chronic periodontitis. Analysis of the T. denticola genome revealed a gene orthologous with a cysteine protease-encoding gene from Streptococcus pyogenes (IdeS). IdeS interferes with IgG-dependent opsonophagocytosis by specific cleavage of IgG molecules. Analysis of this gene (termed ideT) revealed it to encode a two-domain protein whose N-terminus is composed of tandem immunoglobulin-like domains followed by a C-terminal IdeS-like protease domain. In this study we show that during secretion the IdeT protein is processed into an N-terminal fragment which remains associated with the cell, and a C-terminal part released into the medium. Although the secreted domain of IdeT, termed dentipain, shows only 25% identity to the IdeS protease, the putative catalytic cysteine and histidine residues are strongly conserved. Recombinant dentipain cleaves the insulin β-chain, an activity which is inhibited by E-64, a diagnostic inhibitor of cysteine proteases. Apart from insulin no cleavage of other protein substrates was detected, suggesting that dentipain has oligopeptidase activity. A mutant strain was constructed expressing a modified IdeT variant, the dentipain domain of which was deleted. This strain was found to be significantly reduced in its abscess-forming activity compared with the parental strain in a murine abscess model, suggesting that dentipain contributes to the virulence of T. denticola. [ABSTRACT FROM AUTHOR]

Galectins, a family of animal lectins, are involved not only in development and differentiation but also in immunoregulation and host–pathogen interactions. Galectin-3 interacts with lipopolysaccharides in gram-negative bacteria such as Escherichia coli, Salmonella minnesota and Pseudomonas aeruginosa. The present study investigated whether galectin-3 inhibited the cytokine-inducing activity of periodontopathic bacterial lipopolysaccharides using splenocytes derived from mice of different ages. Lipopolysaccharides were extracted from Aggregatibacter actinomycetemcomitans Y4 and Porphyromonas gingivalis ATCC 33277, and then purified. Enzyme-linked immunosorbent assay (ELISA) analysis revealed that galectin-3 adhered to A. actinomycetemcomitans lipopolysaccharides, but not to the lipopolysaccharides of P. gingivalis. Splenocytes were prepared from 1- or 7-month-old C57BL/6 mice. Either A. actinomycetemcomitans lipopolysaccharides (200 ng mL−1) alone or lipopolysaccharides and murine galectin-3 (10 μg mL−1) were added to culture solutions, and the release of interleukin-6 (IL-6) and interferon-γ (IFNγ) from splenocytes was measured by ELISA after a 17-h incubation. In all mice tested, A. actinomycetemcomitans lipopolysaccharide stimulation significantly increased the production of IL-6 and IFNγ ( P<0.01). Murine galectin-3 suppressed lipopolysaccharide-induced cytokine production in the splenocytes of the 1-month-old mice ( P<0.02 for IL-6; P<0.05 for IFNγ), but not in the splenocytes of the 7-month-old mice. This suggests that responses change with age. [ABSTRACT FROM AUTHOR]

Invasion by Porphyromonas gingivalis has been proposed as a possible mechanism of pathogenesis in periodontal and cardiovascular diseases. Porphyromonas gingivalis have direct access to the systemic circulation and endothelium in periodontitis patients by transient bacteremia. Periodontitis can be described as one of the predominant polymicrobial infections of humans. In the present study, P. gingivalis strains were tested for their ability to invade a human gingival epithelial cell line (Ca9-22) and human aortic endothelial cells in coinfection with Fusobacterium nucleatum using antibiotic protection assays. Coinfection with F. nucleatum resulted in 2–20-fold increase in the invasion of host cells by P. gingivalis strains. The invasive abilities of P. gingivalis strains were significantly greater when incubated with a F. nucleatum clinical isolate (which possesses strong biofilm-forming ability), than when incubated with a F. nucleatum-type strain. In inhibition assays with metabolic inhibitors, a difference in inhibition profiles was observed between mono- and polymicrobial infections. Collectively, our results suggest that F. nucleatum facilitates invasion of host cells by P. gingivalis. Investigations of polymicrobial infection of host cells should improve our understanding of the role of P. gingivalis in periodontal infection and proatherogenic mechanisms. [ABSTRACT FROM AUTHOR]

In the previous report of the Prognosis and Statistical Investigation Committee of the Japanese Skin Cancer Society, we tabulated data on patients with malignant melanoma who had been registered at major medical institutions (22 institutions on average) in Japan over 5-year periods from 1987 to 1991 (group A) and from 1992 to 1996 (group B)... [ABSTRACT FROM AUTHOR]

Simvastatin acid (SVA) has been reported to stimulate bone formation by increasing expression of BMP-2 in osteoblasts. Due to their multi-functional characteristics and bioadaptability, cyclodextrins (CDs) are capable of forming inclusion complexes with many drugs by including a whole drug molecule inside their cavity. In the present study, we prepared SVA/CD inclusion complex solutions with different pH values. These were then used to determine their SVA release behavior after coating on titanium substrates, as well as to clarify the characteristics of SVA/CD complexes per se. Results showed that the lower the pH value of the solution, the lower the release kinetics of SVA. Besides, the amount of crystalline complexes in the coatings increased with decrease in pH. These results suggested that the release rate of SVA depended on two factors: pH of the solution and concomitant crystallinity of the coating. [ABSTRACT FROM AUTHOR]

We studied the effect of antibodies against Porphyromonas gingivalis gingipain domains, preparing them against three recombinant fragments of RgpA (catalytic domain, r-Rgp CAT; hemagglutinin domains, r-Rgp 44 and r-Rgps 15–27) and one fragment of Kgp (catalytic domain, r-Kgp CAT). Enhancement of opsonization and killing by human polymorphonuclear leukocytes were measured in the noninvasive FDC 381 and invasive W50 strains of P. gingivalis. Anti-r-Rgp 44 was the most effective in both strains of P. gingivalis. The present findings lead us to recommend RgpA 44 as a candidate immunogen for vaccines against P. gingivalis. [ABSTRACT FROM AUTHOR]

The article discusses research on the relation of periodontopathic anaerobic bacteria to aspiration pneumonia. Topics include the formation of periodontopathic biofilms in elderly people, the coexistence of Porphyromonas gingivalis and Treponema denticola, and the use of professional oral health care (POHC) to decrease anaerobic bacteria that cause aspiration pneumonia.

Increasing evidence has linked the anaerobic bacteria forming periodontopathic biofilms with aspiration pneumonia in elderly persons. In experiments designed to eliminate the potent respiratory pathogens forming biofilms in the oral cavity, we have shown that the mechanical and chemical oral cleansing using povidone-iodine effectively reduced the detection rates and numbers of methicillin-sensitive Staphylococcus species, Streptococcus pneumoniae, and Haemophilus influenzae in patients scheduled to undergo oral surgery requiring endotracheal intubation. We confirmed the pathogenicity of periodontopathic anaerobic bacteria for aspiration pneumonia in an experimental mouse model. Based upon the finding of the co-existence of Porphyromonas gingivalis with Treponema denticola in chronic periodontitis lesions, we innoculated a mixed culture of P. gingivalis and T. denticola into the mouse trachea; the resulting infection induced inflammatory cytokine production and caused pneumonia. In another series of investigations, professional oral health care (POHC), mainly cleansing administered by dental hygienists once a week for 24 months to elderly persons requiring daily care, resulted in the reduction of the number of total anaerobes, Candida albicans, and Staphylococcus species and in the number of eases of fatal aspiration pneumonia. We also found that the POHC treatment of elderly persons for 6 months in the winter season reduced the salivary levels of protease, trypsin-like activity, and neuraminidase and also decreased the frequency of influenza cases. [ABSTRACT FROM AUTHOR]

A protease of Treponema denticola, dentilisin, is thought to be part of a complex with 43- and 38-kDa proteins. A sequence encoding a 43-kDa protein was located in the 3′ region of the prcA gene upstream of the dentilisin gene (prtP). The 43-kDa protein was apparently generated from digestion of PrcA. To clarify the function of the protein, we constructed a mutant of the 43-kDa protein following homologous recombination. The mutant lacked detectable dentilisin activity. Immunoblot analysis demonstrated that the dentilisin protein was degraded in the mutant. The results of real-time polymerase chain reaction suggested that prtP mRNA expression in the mutant was somewhat decreased compared with the wild-type strain. These data suggest that the 43-kDa protein is involved in the stabilization of the dentilisin protein. [Copyright &y& Elsevier]

Background: Arginine- and lysine-specific cysteine proteinases (arg-gingipain: Rgp, lys-gingipain: Kgp) are major virulence factors of Porphyromonas gingivalis. Recent reports have suggested that antibodies against gingipains can play a protective role against infection by P. gingivalis. The purpose of this study was to evaluate the IgG responses of patients with periodontitis to functional domains of gingipains.Methods: A group of 29 periodontitis patients and 10 periodontally healthy subjects (control group) were recruited into this study. We prepared three recombinant fragments of rgp A (catalytic domain; r-Rgp CAT) and two hemagglutinin domains (r-Rgp 44, and r-Rgps 15-27) corresponding to amino acid residues 228 to 719, 720 to 1136, and 1137 to 1704, respectively. One fragment of the Kgp catalytic domain (r-Kgp CAT) corresponding to amino acid residues 229 to 737 and expressed in Escherichia coli was also used. IgG antibody levels to these recombinant proteins in sera from the subjects were determined by an enzyme-linked immunosorbent assay (ELISA).Results: We found that IgG levels against r-Rgp 44 and r-Rgps 15-27 in sera obtained from the patients were significantly higher than those in the healthy group (P<0.01). In contrast, no significant differences in IgG levels against r-Rgp CAT and r-Kgp CAT were found between the control and patient groups. The IgG responses to P. gingivalis sonic extracts, r-Rgp 44 and r-Rgps 15-27, were related to probing depth in sera from patients, but those to r-Rgp CAT and r-Kgp CAT were not.Conclusion: The present findings suggest that the low responsiveness of IgG antibody against the catalytic domains of gingipain, r-Rgp CAT, and r-Kgp CAT is a key factor in infection by P. gingivalis. [ABSTRACT FROM AUTHOR]

Background: Porphyromonas gingivalis is one of the major microbial pathogens associated with chronic periodontitis. To eradicate such pathogens by periodontal therapy, it is essential to clarify the source of infection. Recent findings suggest that the genotype of the fimbriae is one of the important factors in infection by P. gingivalis. The objectives of the present study were to investigate the transmission of P. gingivalis between spouses and to determine the relationship between P. gingivalis fimA type and colonization. Methods: A total of 14 couples were selected to investigate the transmission of P. gingivalis and its association with the fimA types. To examine the distribution of fimA type in the general population. 32 subgingival plaque samples from 47 patients with periodontitis were also tested. The transmission of P. gingivalis strains was determined by using pulsed field gel electrophoresis (PFGE). P. gingivalis strains isolated from the couples and subgingival dental plaque samples were studied for fimA classification. Results: The PFGE patterns of P gingivalis strains from matched husbands and wives were identical for six of the 14 couples. In five of these six couples (B3.3%). P. gingivalis strains harboring the type II fimA gene were present. The proportion of type II fimA in the strains isolated from couples with probable intrafamilial transmission was significantly higher than that in patients with periodontitis or in the group of samples isolated from one member of a couple Conclusion: This study suggests that fimA type II, even though widely distributed in patients with periodontitis, may be an important factor in the transmission of P. gingivalis between spouses. [ABSTRACT FROM AUTHOR]

.Purpose. This study was conducted to evaluate the usefulness of mechanical and chemical prophylactic oral cleansing treatments for reducing potential respiratory pathogens existing in the oral cavity.Methods. Thirty-two patients scheduled to undergo oral and maxillofacial surgery that required endotracheal anesthesia were randomly allocated to one of the two groups, the oral cleansing group (n = 16) or the noncleansing group (n = 16). Culture and polymerase chain reaction (PCR) methods were used to detect and enumerate pathogens. Oral cleansing was carried out with an electric toothbrush capable of automatically supplying and aspirating povidone-iodine solution before surgery, followed by rinsing twice a day after surgery. Cephazolin (3 g·day[sup -1] ) was given to all patients for 5 days after surgery.Results. The PCR detection rates of Streptococcus pneumoniae, Haemophilus influenzae, Pseudomonas aeruginosa, and Porphyromonas gingivalis in gargle samples before treatment were 87.5%, 68.8%, 53.1%, and 40.6%, respectively. Oral cleansing reduced the detection rates and numbers of methicillin-sensitive Staphylococcus species, S. pneumoniae, and H. influenzae. In contrast, there was no significant reduction of methicillin-resistant Staphylococcus species, S. pneumoniae, H. influenzae, or P. aeruginosa in subjects who underwent systemic cephazolin administration without oral cleansing.Conclusion. The combination of mechanical and chemical oral cleansing resulted in a significant reduction of potential respiratory pathogens in the oral cavity. [ABSTRACT FROM AUTHOR]

Campylobacter rectus, a gram-negative, microaerophilic, and motile bacterium, has been proposed to play a pathogenic role in human periodontitis. Surface components, such as the flagellum, surface layer (S-layer), and cytotoxin, have been reported as possible virulence factors of the microorganism. In the present study, monoclonal antibodies against surface components of this bacterium were produced to detect and investigate the pathogenic potential of C. rectus in periodontitis. Two monoclonal antibodies, designated CRT-1 and CRT-2, recognized a peculiar 150 kDa S-layer protein by immunoblot analysis. The CRT-2 antibody reacted to all C. rectus strains tested, except for the S-layer negative strain of the species [C. rectus ATCC 33238 S-laver (-) strain]. The CRT-3 antibody reacted to a 60-kDa protein in C. rectus and also cross-reacted with Campylobacter showae ATCC 51164 and CCUG 11641 strains. Using the dot-blot method, we were able to detect C. rectors using the CRT-2 antibody when as few as 10³ organisms were present in a subgingival dental plaque sample. Detection of C. rectors in plaque samples correlated significantly with clinical findings such as probing depth (P < 0.001), bleeding on probing (P < 0.001), and gingival index (P < 0.001). These findings indicate that infection by C. rectus may be an important indicator of periodontal disease status. [ABSTRACT FROM AUTHOR]

Increasing evidence has linked colonization by Helicobacter pylori with the development of gastritis and peptic ulcer disease. H. pylori resides primarily in the gastric mucosa without invading the gastric epithelium, causing persistent mild gastric inflammation. There are many reports examining the relationship between colonization by microorganisms in the stomach and oral cavity. We found that some oral bacteria are able to trap H. pylori cells, but oral bacteria inhibit H. pylori growth in vitro. In cases where H. pylori was detected in oral cavity samples, including oral cancer surface samples, we suggested that this species had colonized the stomach and were present in the oral cavity only as a transient organism. We demonstrated that periodontopathic Campylobacter rectus strains posses proteinaceous antigens, including heat shock proteins that share antigenicity with antigens of H. pylori strains. These cross-reactive antigens between H. pylori and C rectus may be related to the induction of immunopathological responses in periodontal tissues and the stomach. We concluded that H. pylori could not survive in the human oral cavity; however, there would be an interrelationship between periodontal disease due to C. rectux and stomach diseases due to H. pylori. [ABSTRACT FROM AUTHOR]

Background: Periodontopathic bacteria induce inflammation of periodontal tissues. The cytokines and nitric oxide released in periodontal lesions have been reported to play a protective role in bacterial infection and to relate lo the process of inflammation. To clarify the relationship between colonization of periodontopathic bacteria and cytokines, we evaluated profiles of inflammatory cytokines, chemokine, anti-inflammatory cytokines. and inducible nitric oxide synthase (iNOS) and colonization by Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans, which are major pathogens of periodontitis. Methods: mRNA expression of cytokines and iNOS in inflamed and healthy gingival tissue was determined using reverse transcription-polymerase chain reaction (RT-PCR), and the relationship between their profiles and the detection of specific bacteria was analyzed. Results: The relative expression of interleukin (IL)-6 and iNOS mRNAs in periodontal lesions was significantly higher than those in healthy individuals. IL-6 mRNA expression was also significantly higher at bleeding on probing (BOP)-positive sites than at BOP-negative sites. The expressions of IL-1α and IL-8 increased, but IL-10 expression decreased at sites where A. actinomycetemcomitans was detected. We found no correlation between the expression of cytokine and iNOS mRNA and infection by P. gingivalis. Conclusions: The expression of IL-6 may reflect inflammation in gingival tissue, and iNOS may be involved in the inflammatory process in periodontitis. The presence of A. actinomycetemcomitans or P. gingivalis might relate to the different cytokine profiles of IL-1α, IL-8, and IL-10. [ABSTRACT FROM AUTHOR]

Cytogenetic studies were performed in a 72-year-old male patient with triple primary cancers including breast, skin and lung. Left breast cancer was diagnosed at the age of 46 and he received mastectomy and thoracic irradiation. Squamous cell carcinoma and Bowen's disease were diagnosed from two separated parts of a skin lesion at the age of 70. Small-cell lung cancer was diagnosed 1 year later, and he received chemotherapy and radiotherapy. Chromosome analysis was carried out on both peripheral lymphocyte and skin fibroblast cultures at the age of 72. Out of 30 fibroblast cells karyotyped at the second passage, 7 cells (23%) consistently showed a reciprocal translocation t(Y;6)(q12;p21). The same translocation was found in one of 200 cells from lymphocyte cultures. The findings suggest that the translocation t(Y;6) might be inherent in nature, and that the patient was a mosaic of 46,XY/ 46,X,t(Y;6)(q12;p21). These results highlight the constitutional chromosomal abnormality as one of the possible high-risk factors for multiple primary cancers. [ABSTRACT FROM AUTHOR]

Chloramphenicol-resistant (CAP) reconstituted cells and cybrids were isolated by fusion of karyoplasts (or intact cells) of mouse amelanotic melanoma B cells with cytoplasts of hypoxanthine-guanine phosphoribosyltransferase (HGPRT)-deficient, CAP rat myoblastic cells, LTG.CAP, and double selection in HAT medium containing CAP. Reconstituted cells or cybrids exhibited unique cellular arrangement, and about one third of the isolated clones expressed high tyrosinase activity and marked melanin synthesis, although the parental mouse cells expressed low tyrosinase activity and the parental rat cells did not express tyrosinase activity. These phenotypic changes have been stable for more than a year. The phenotypic reversions of these clonal cells were induced by treatment with a tumor promoter. There were changes in the morphology of the treated cells to that of the mouse B cells and extinction of tyrosinase activity and melanin synthesis in pigmented clonal cells. These phenotypic changes and reversions induced by a promoter were repeatedly reversible. [ABSTRACT FROM AUTHOR]

The adherence of hpopolysaccharides (LPSs) from periodontal disease-associated bacteria to saliva-coated hydroxyapatite (S-HA) and serum-coated HA beads was examined by chromogenic Limulus activity (toxicolor test). Phenol-water extracted LPS preparations from Bacteroides intermedius, Fusobacterium nucleatm, Actinobacillus actinomycetemcomitans, Eikenella corrodens, and rough-type LPS from Escherichia coli adhered to S-HA and serum-coated beads and agglutinated human erythrocytes. The adhered LPSs to S-HA and serum-coated HA beads were not removed by vigorous washing with distilled water LPSs from Bacteroides (Por phyromonas) gingivalis strains and wild-type E. coli did not adhere to S-HA, Serum-coated HA beads or show hemagglutinating activity SDS-PAGE patterns stained with silver stain showed that LPSs adhered to S-HA, and serum-coated HA beads and erythrocytes possessed a distinct fast-migrating band similar to rough-type LPS B. gingivalis LPSs possessed slow-migrating and repeating ladder bands similar to wild-type LPS [ABSTRACT FROM AUTHOR]

THE PURPOSE OF THIS STUDY WAS TO EXAMINE avidity of immunoglobulin G (IgG) antibody for surface antigens of Porphyromonas gingivalis in sera from patients with adult periodontitis. The antigens used were whole cell antigens, lipopolysaccharides (LPS), and fimbriae of non-invasive P. gingivalis ATCC 33277 and invasive 16-1. Serum IgG titers for the P. gingivalis antigens were measured by enzyme-linked immunosorbent assay (ELISA) before and after periodontal initial preparation. IgG avidity was measured by diethylamine dissociation ELISA. IgG titers for the whole cell antigens of 16-1, LPS, and the fimbria antigens from both P. gingivalis strains were significantly higher in the patient group than those in the control group, whereas values for avidities were significantly lower in patient sera. Although we found a statistically significant decrease in the IgG titers of patients following initial preparation, avidities against the fimbria antigen of invasive 16-1 strain increased. The present study showed that IgG antibodies elicited in patients were different in their ability to respond to invasive P. gingivalis 16-1 and to non-invasive 33277. The patient sera with high IgG titers demonstrated low values for avidity, suggesting that IgG responses in patients play a limited role in colonization inhibition or elimination of P. gingivalis. The data indicate that periodontally healthy individuals may have highly functional antibodies which may protect against P. gingivalis colonization. Our findings suggest that the ability to produce functional antibodies in the patient group is lower than that in the periodontally healthy group, but the functional antibodies can be induced by the initial preparation. [ABSTRACT FROM AUTHOR]

An enzymatic method, SK-013, was developed for rapid detection of the peptidase activity in subgingival plaque samples. This method was found to have specificity for Porphyromonas gingivalis, Treponema denticola, Bacteroides forsythus, and some Capnocytophaga strains. The purpose of this study was to determines whether SK-013 could indicate the presence of periodontopathic bacteria including T. denticola, P. gingivalis and B. forsythus, which produce trypsin-lime enzymes. Subgingival plaque samples were taken from 10 clinically healthy sites and 30 periodontally diseased sites with 3 paper points. SK-013 activity of plaque samples was assayed, and the numbers of T. denticola, P. gingivalis and B. forsythus in the sample were counted by immunofluorescence technique. In diseases sites, the SK-013 activity was significantly correlated with clinical parameters such as Gingival Index. Plaque Index, probing depth and bleeding on probing. A significant correlation was found between the presence of these organisms and SK-013 activity. Correlation coefficients between the presence of T. denticola and SK-013 activity were higher than those with other organisms. These findings indicate that the SK-013 is useful as an indicator of cell population of T. denticola, P. gingivalis and B. forsythus in subgingival plaque. [ABSTRACT FROM AUTHOR]

Porphyromonas gingivalis, Bacteroides forsythus, and Treponema denticola have been found to predominate in peridontal pockets of patients with adult periodontitis. These microorganisms hydrolyze the synthetic peptide N-benezoyl-DL-arginine-2-napththylamide (BANA). In this study, we developed an enzymatic method , designated SK-013, to detect the existence of these microorganisms in subgingival plaque bacteria. This enzymatic method was based on the observation of the hydrolysis of N-carbobenzoxy-glycyl-glycyl-arginyl-3,5-dibromo-4-hydroxyaniline (N-CBz-GLy-ARg-DBHA) and made more sensitive by adding an enhancing system. The SK-013 was specifically positive for P. gingivalis, B. forsythus. T. denticola, and some strains of Capnocytophaga species, but was not specific for any of the other bacterial strains tested. This SK-013 system may be valuable for detection and quantification of periodontal disease-associated bacteria in sub-gingival plaque and thus for diagnosis of peridontal infections. [ABSTRACT FROM AUTHOR]

Periodontitis is an inflammation of tooth-supporting tissues, which is caused by bacteria in the subgingival plaque (biofilm) and the host immune response. Traditionally, subgingival pathogens have been investigated using methods such as culturing, DNA probes, or PCR. The development of next-generation sequencing made it possible to investigate the whole microbiome in the subgingival plaque. Previous studies have implicated dysbiosis of the subgingival microbiome in the etiology of periodontitis. However, details are still lacking. In this study, we conducted a metagenomic analysis of subgingival plaque samples from a group of Japanese individuals with and without periodontitis. In the taxonomic composition analysis, genus Bacteroides and Mycobacterium demonstrated significantly different compositions between healthy sites and sites with periodontal pockets. The results from the relative abundance of functional gene categories, carbohydrate metabolism, glycan biosynthesis and metabolism, amino acid metabolism, replication and repair showed significant differences between healthy sites and sites with periodontal pockets. These results provide important insights into the shift in the taxonomic and functional gene category abundance caused by dysbiosis, which occurs during the progression of periodontal disease. [ABSTRACT FROM AUTHOR]

A total of 8 strains of 6 bacterial species, Porphyromonas gingivalis ATCC33277 and TDC286, Actinobacillus actinomycetemcomitans ATCC29523 and JP2, Fusobacterium nucleatum No. 2, Tannerella forsythensis ATCC43937, Prevotella intermedia ATCC25611 and Streptococcus anginosus ATCC33397, were treated with povidone-iodine (PVP-I) gargle (PVP-I: 0.47 and 0.23% w/v) or chlorhexidine gluconate (CHG) gargle (CHG: 0.002% w/v) for 15, 30 or 60 s, after which they were inoculated into various media, cultured and counted for residual bacteria. At both concentrations, PVP-I gargle reduced the viable cell count of all 8 bacterial strains to below the measurable limit within 15 s. By contrast, there were more than 1,000 viable colonies 60 s following treatment with the CHG gargle. The results demonstrate that povidone-iodine gargle has rapid bactericidal activity against the causative bacteria of periodontal disease. Copyright © 2006 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

In order to find biomarkers to measure the effects of UV irradiation, we examined the accumulation of p53 protein and pyrimidine dimers in 18 solar keratosis specimens. Frozen or AMeX-fixed solar keratosis specimens were immunohistochemically stained by anti-p53 mouse monoclonal antibody, pAb1801 and polyclonal anti-(pyrimidine dimer) antibody. Nuclear accumulation of p53 protein was found in 5/18 (28%) solar keratosis lesions. The percentage of cases showing nuclear p53 protein varied according to the histological type; in the bowenoid type it was 4/7 (57%); in the atrophic type it was 1/7 (14%). Nuclear pyrimidine dimers were not stained in solar keratosis, although the skin of UV-irradiated nude mice was positive. Accumulation of p53 protein is a good marker for early precancerous change caused by UV exposure. [ABSTRACT FROM AUTHOR]