p. gingivalis Search Results


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  • 99
    ATCC p gingivalis
    Survival curve of G. mellonella infected with P. <t>gingivalis</t> at different concentrations of pomegranate extract. The concentrations of PGE (12.5 mg/mL, 6.25 mg/mL, 3.1 mg/mL, and 2.5 mg/mL) diluted in PBS were inoculated in the last proleg of each larva. Fifteen larvae were used per group. The number of dead G. mellonella was daily recorded for 168 hours for analysis of the survival curve. The larvae survived at concentrations of 12.5 mg/mL, 6.25 mg/mL, 3.1 mg/mL, and 2.5 mg/mL.
    P Gingivalis, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 2130 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Astarte Biologics p gingivalis
    Survival curve of G. mellonella infected with P. <t>gingivalis</t> at different concentrations of pomegranate extract. The concentrations of PGE (12.5 mg/mL, 6.25 mg/mL, 3.1 mg/mL, and 2.5 mg/mL) diluted in PBS were inoculated in the last proleg of each larva. Fifteen larvae were used per group. The number of dead G. mellonella was daily recorded for 168 hours for analysis of the survival curve. The larvae survived at concentrations of 12.5 mg/mL, 6.25 mg/mL, 3.1 mg/mL, and 2.5 mg/mL.
    P Gingivalis, supplied by Astarte Biologics, used in various techniques. Bioz Stars score: 92/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    InvivoGen p gingivalis
    The effect of AEA on the production of pro-inflammatory mediators in hPdLCs in response to stimulation with P. <t>gingivalis</t> LPS. hPdLC were stimulated with P. gingivalis LPS in the presence or in the absence of AEA for 24 h, and the production of pro-inflammatory mediators was measured on gene and protein levels by real-time PCR (A) and ELISA (B) respectively. A – Changes in the gene expression levels of IL-6, IL-8, and MCP-1 calculated by 2 −ΔΔCt method taking non-stimulated cells as a reference (2 −ΔΔCt = 1) and β-actin as a house-keeping gene. B – Content of pro-inflammatory mediators in the conditioned media measured by ELISA. Each value represents the mean ± s.e.m of 4 independent experiments. # Means were significantly compared to the control group, and tested with analysis of variance (P
    P Gingivalis, supplied by InvivoGen, used in various techniques. Bioz Stars score: 95/100, based on 317 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher p gingivalis
    Chemokines and cytokines secreted in HCAEC stimulated with P. <t>gingivalis</t> -LPS or live- P. gingivalis . Monolayers of HCAEC cultured in 12-well plates were stimulated with P. gingivalis -LPS (1.0, 3.5, 7.0 µg/mL) or P. gingivalis (MOI 1:100 - 1:0,1) for 24 h under repeated exposure (+++) or single exposure (+). After stimulation, levels of the following chemokines were measured in cell culture supernatants using a cytometric bead array: ( A ) IL-8, ( B ) MCP-1, ( C ) IL-6, ( D ) GM-CSF, ( E ) IL-1β. Symbol (*) means p
    P Gingivalis, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 369 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    DSMZ p gingivalis
    Effects of OMV preexposure on TNF secretion in response to secondary stimulation with live P. <t>gingivalis</t> . (A) Human monocytes were prestimulated or not prestimulated with OMV for 24 h; after washing, cells were left unstimulated (medium) or restimulated
    P Gingivalis, supplied by DSMZ, used in various techniques. Bioz Stars score: 93/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    InvivoGen p gingivalis lps
    P. <t>gingivalis</t> <t>LPS-induced</t> Wnt5a expression was inhibited by wedelolactone or dnIκBα. (A, B) THP-1 cells were pre-treated with the IKK inhibitor wedelolactone or the IKK inhibitor VII for 1 hr. The expression of Wnt5a mRNA was detected by real-time PCR. Relative expression levels of Wnt5a are presented. DMSO is a solvent control for both inhibitors. (C) THP-1 cells were transfected with a dominant-negative form of IκBα (dnIκBα) or the parent plasmid (pUSE) for 12 hrs. Fold induction of Wnt5a mRNA expression is shown. *p
    P Gingivalis Lps, supplied by InvivoGen, used in various techniques. Bioz Stars score: 95/100, based on 155 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher p gingivalis lps
    Cytokine expression levels upon stimulation of U937 macrophages with E. coli (0.1 μg/ml), P. <t>gingivalis</t> , and T. forsythia (1 μg/ml) <t>LPS</t> in the presence or absence of FCS. In both qPCR (A) and ELISA (B), P. gingivalis LPS did not show serum dependence for cytokine production. E. coli and T. forsythia LPS caused a substantially higher release of IL-1β, IL-6, and TNF-α in the presence of serum. The data are mean values ± SDs of three wells originating from one representative experiment. A similar tendency was observed in other experiments. *, significantly different from the control at P
    P Gingivalis Lps, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Gene Codes Inc p gingivalis w83
    Cytokine expression levels upon stimulation of U937 macrophages with E. coli (0.1 μg/ml), P. <t>gingivalis</t> , and T. forsythia (1 μg/ml) <t>LPS</t> in the presence or absence of FCS. In both qPCR (A) and ELISA (B), P. gingivalis LPS did not show serum dependence for cytokine production. E. coli and T. forsythia LPS caused a substantially higher release of IL-1β, IL-6, and TNF-α in the presence of serum. The data are mean values ± SDs of three wells originating from one representative experiment. A similar tendency was observed in other experiments. *, significantly different from the control at P
    P Gingivalis W83, supplied by Gene Codes Inc, used in various techniques. Bioz Stars score: 86/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Luminex p gingivalis culture
    Cytokine expression levels upon stimulation of U937 macrophages with E. coli (0.1 μg/ml), P. <t>gingivalis</t> , and T. forsythia (1 μg/ml) <t>LPS</t> in the presence or absence of FCS. In both qPCR (A) and ELISA (B), P. gingivalis LPS did not show serum dependence for cytokine production. E. coli and T. forsythia LPS caused a substantially higher release of IL-1β, IL-6, and TNF-α in the presence of serum. The data are mean values ± SDs of three wells originating from one representative experiment. A similar tendency was observed in other experiments. *, significantly different from the control at P
    P Gingivalis Culture, supplied by Luminex, used in various techniques. Bioz Stars score: 94/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Becton Dickinson p gingivalis
    Anti-microbial activity of clinical working concentrations of PHMG-P (1%) and CHX (0.2%) against perio- and cariogenic bacteria ( P. <t>gingivalis</t> , A. actinomycetemcomitans , S. mutans and L. acidophilus ).
    P Gingivalis, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 90 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Difco p gingivalis
    Persistence of P. <t>gingivalis</t> within HCAE cells over 8 h. HCAE cells were preincubated for 1 h at 37°C with either fresh antibiotic-free EGM-2 (solid bars), 10 mM 3-methyladenine (open bars), or 10 nM wortmannin (checkered bars) ( n = 3). P. gingivalis 381 cells were incubated with EGM-2 with or without the appropriate inhibitor. The number of CFU of HCAE cells infected with P. gingivalis in the absence of autophagy inhibitors grew over the 8 h, whereas the number of CFU of HCAE cells infected with P. gingivalis in the presence of either autophagy inhibitor declined over the 8 h. The number of CFU for E. coli MC1061, the negative control, at 2.5 h postinfection was
    P Gingivalis, supplied by Difco, used in various techniques. Bioz Stars score: 89/100, based on 52 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Illumina Inc p gingivalis
    Persistence of P. <t>gingivalis</t> within HCAE cells over 8 h. HCAE cells were preincubated for 1 h at 37°C with either fresh antibiotic-free EGM-2 (solid bars), 10 mM 3-methyladenine (open bars), or 10 nM wortmannin (checkered bars) ( n = 3). P. gingivalis 381 cells were incubated with EGM-2 with or without the appropriate inhibitor. The number of CFU of HCAE cells infected with P. gingivalis in the absence of autophagy inhibitors grew over the 8 h, whereas the number of CFU of HCAE cells infected with P. gingivalis in the presence of either autophagy inhibitor declined over the 8 h. The number of CFU for E. coli MC1061, the negative control, at 2.5 h postinfection was
    P Gingivalis, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 85/100, based on 77 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Nissui Pharmaceutical p gingivalis
    Time-kill curves for P. <t>gingivalis</t> JCM12257 (a) and A. actinomycetemcomitans JCM8577 (b) following exposure to 0.9% NaCl (control), 0.2% CHX, and NBW3. The number of CFUs/mL of P. gingivalis exposed to 0.2% CHX did not drop to below the lower limit of detection (
    P Gingivalis, supplied by Nissui Pharmaceutical, used in various techniques. Bioz Stars score: 90/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Promega p gingivalis
    fim region able to swap during natural competence despite limited conservation regions. (A.) Complete fim gene region ( fimA-fimE ) and flanking genes are shown for sequenced P. <t>gingivalis</t> strains 33277 fimA (I ), TDC60 fimA (II ), and W83 fimA (IV ). Overall sequence conservation between all strains is displayed. Green indicates the most conservation while yellow, orange, and finally red signifies the least DNA conservation between strains. Distances are shown in base pairs. Figure modified from CLC Genomic Workbench 6, accession numbers 33277(NC_010729), TD60 (NC_015571), and W83 (NC_002950). (B.) Chromosomal preparations from a random sample of ten recovered recombinants (#1–10) from the dead donor assay (donor W83, recipient 33277) were tested for the presence of the donor fimA type IV allele using PCR amplification. In addition, to determine if regions further downstream of fimA were also swapped, the recombinants were tested for the presence of donor fimC, fimD , and fimE . ‘ X ’ indicates the presence of the donor gene and ‘ O ’ indicates the presence of the recipient gene in the tested strains.
    P Gingivalis, supplied by Promega, used in various techniques. Bioz Stars score: 94/100, based on 97 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Zambon p gingivalis
    fim region able to swap during natural competence despite limited conservation regions. (A.) Complete fim gene region ( fimA-fimE ) and flanking genes are shown for sequenced P. <t>gingivalis</t> strains 33277 fimA (I ), TDC60 fimA (II ), and W83 fimA (IV ). Overall sequence conservation between all strains is displayed. Green indicates the most conservation while yellow, orange, and finally red signifies the least DNA conservation between strains. Distances are shown in base pairs. Figure modified from CLC Genomic Workbench 6, accession numbers 33277(NC_010729), TD60 (NC_015571), and W83 (NC_002950). (B.) Chromosomal preparations from a random sample of ten recovered recombinants (#1–10) from the dead donor assay (donor W83, recipient 33277) were tested for the presence of the donor fimA type IV allele using PCR amplification. In addition, to determine if regions further downstream of fimA were also swapped, the recombinants were tested for the presence of donor fimC, fimD , and fimE . ‘ X ’ indicates the presence of the donor gene and ‘ O ’ indicates the presence of the recipient gene in the tested strains.
    P Gingivalis, supplied by Zambon, used in various techniques. Bioz Stars score: 91/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Novoprotein p gingivalis
    Transmission electron micrographs demonstrating P. <t>gingivalis</t> invasion of EC. (A and B) BAEC with P. gingivalis A7436. (A) At the cell surface, bacteria appear to induce EC structural rearrangements consistent with an endocytic mechanism. (B) Internalized bacteria are found within vacuole. (C) BAEC with P. gingivalis 381. Note the apparent contact between microfilamentous cellular components and surface-adhering P. gingivalis . (D) BAEC with P. gingivalis fimA mutant DPG3. Absence of intimate interaction between EC surface and bacteria. (E and F) FBHEC incubated with P. gingivalis A7436. Surface adherence (E) and engulfment in vacuole (F). Arrows in all panels point to P. gingivalis . Bars on each image are 0.5 μm unless otherwise specified. Composite image was constructed with Adobe Photoshop 3.0.
    P Gingivalis, supplied by Novoprotein, used in various techniques. Bioz Stars score: 81/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    TaKaRa p gingivalis
    The effects of S. mitis , E. coli , heat-killed P. <t>gingivalis</t> and the supernatant prepared from P. gingivalis cultures on the microglial response. ( a–l ) The dynamic response of the microglial processes to the local injection of various bacteria (3.6 × 10 5 CFU ml −1 ) at ZT2. ( a–c ) S. mitis (S.m), ( d–f ) E. coli (E.c), ( g–i ) heat killed P.g (HK-P.g; 95 °C, 10 min), ( j–l ) P.g culture supernatant (P.g SN). Scale bar: 10 μm. ( m ) The kinetics of the mean fluorescent change in the microglial response measured at 30 min after the injection of various bacteria. The data are presented as the mean ± S.E.M. (N = 3 mice, n = 4–5 each). A one-way ANOVA with a post hoc Tukey’s test; P.g vs S.m: p > 0.9999, Pg. vs. E.c: p = 0.9953, P.g vs. HK-P.g: p = 0.4244, S.m vs. E.c: p = 0.9939, S.m vs. HK-P.g: p = 0.4126, E.c vs. HK-P.g: p = 0.5521. ( n ) The kinetics of the mean fluorescent change in the microglial response measured at 30 min after the injection of P. gingivalis . The data are presented as the mean ± S.E.M. (N = 3 mice, n = 5–6 each). A two-tailed unpaired t -test; P.g vs BHI (P.g culture medium, as a control): p = 0.0856.
    P Gingivalis, supplied by TaKaRa, used in various techniques. Bioz Stars score: 94/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore p gingivalis
    Effects of OMV preexposure on TNF secretion in response to secondary stimulation with live P. <t>gingivalis</t> . (A) Human monocytes were prestimulated or not prestimulated with OMV for 24 h; after washing, cells were left unstimulated (medium) or restimulated
    P Gingivalis, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 208 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies p gingivalis
    LXA 4 inhibits P. <t>gingivalis</t> -induced upregulation of CD11b/CD18 and exposure of the CD11b/CD18 high-affinity epitope on neutrophils. Whole blood was incubated in the presence or absence of LXA 4 (100 or 500 nM) for 15 min and subsequently stimulated with
    P Gingivalis, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 93/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cedarlane p gingivalis
    LXA 4 inhibits P. <t>gingivalis</t> -induced upregulation of CD11b/CD18 and exposure of the CD11b/CD18 high-affinity epitope on neutrophils. Whole blood was incubated in the presence or absence of LXA 4 (100 or 500 nM) for 15 min and subsequently stimulated with
    P Gingivalis, supplied by Cedarlane, used in various techniques. Bioz Stars score: 88/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Hitachi Ltd p gingivalis
    Effect of the Nano-MIX on the multi-species biofilms of S. mutans , F. nucleatum , A. actinomycetemcomitans , and P. <t>gingivalis</t> at 24 h. The confocal scanning laser microscopy (CLSM) ( A , B ) and scanning electron microscopy (SEM) images ( C , D ) showing comparative antibacterial effects of the Nano-MIX treatment ( B , D ) on the mixed-species oral biofilms with reference to the blank nanoparticles ( A , C ), respectively.
    P Gingivalis, supplied by Hitachi Ltd, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    iNtRON Biotechnology p gingivalis
    Immunohistochemical analysis of SDF-1α and CXCR4 in P. <t>gingivalis</t> challenged experimental rat periodontal inflammation. SDF-1α and CXCR4 were abundantly expressed in the P. gingivalis challenged rat tissues. Immunohistochemical analysis revealed a higher expression of SDF-1α and CXCR4 in P. gingivalis challenged rat gingiva (Fig. 4a; Scale bars, 20 μM) and PDL (Fig. 4b; Scale bars, 100 μM) compared to the control
    P Gingivalis, supplied by iNtRON Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Merck KGaA p gingivalis
    Immunohistochemical analysis of SDF-1α and CXCR4 in P. <t>gingivalis</t> challenged experimental rat periodontal inflammation. SDF-1α and CXCR4 were abundantly expressed in the P. gingivalis challenged rat tissues. Immunohistochemical analysis revealed a higher expression of SDF-1α and CXCR4 in P. gingivalis challenged rat gingiva (Fig. 4a; Scale bars, 20 μM) and PDL (Fig. 4b; Scale bars, 100 μM) compared to the control
    P Gingivalis, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 91/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Peptide Institute p gingivalis
    Schematic illustration of extracellular oligopeptide metabolism in P. <t>gingivalis</t> . The metabolic pathway of P. gingivalis from the extracellular polypeptides, di- and tri-peptide incorporation, amino acid metabolism, and excretion as short-chain fatty acids, are schematically illustrated [10] , [15] . Amino acids, except for Ser and Thr, are mainly transported as di- and tri-peptides via oligopeptide transporters [10] . Rgp and Kgp are mainly localized on the outer membrane (OM), while DPPs, PtpA, and AOP are located in periplasmic space [23] , [24] , [25] . Scissors indicate peptidases, which cleave peptide bonds at specific positions. Stars represent acylaminoacyl groups at the N-terminus. IM, inner membrane.
    P Gingivalis, supplied by Peptide Institute, used in various techniques. Bioz Stars score: 87/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    InvivoGen ultrapure p gingivalis lps
    P. <t>gingivalis</t> bypasses the epithelial barrier to reach FBs located at the core of the MT leading to MT destruction. SEM imaging of ( A , B ) P. gingivalis culture, ( C ) P. gingivalis infection and intracellular invasion of the MT surface, ( D ) P. gingivalis proliferating on MT surface and EC exfoliation, ( E ) uninfected MT, ( F ) MT stimulated with <t>LPS,</t> ( G ) MT infected with P. gingivalis for 6 h and ( H ) for 24 h. Green arrows indicate P. gingivalis infection in MT and/or cell invasion; blue arrows indicate the disruption of epithelial barrier, multiple apoptotic cells and an acute cellular injury in all strata of the MT – EC and FB-; white arrow indicates proliferating EC at the surface of MT.
    Ultrapure P Gingivalis Lps, supplied by InvivoGen, used in various techniques. Bioz Stars score: 93/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher ultrapure p gingivalis lps
    (A) IL-6 and TNFα levels in tissue culture supernatants from unstimulated (no <t>LPS)</t> and P. <t>gingivalis</t> LPS-stimulated cell lines, and (B) unstimulated (no LPS) and E. coli LPS-stimulated cell lines. (C) Anti-TLR4 antibody suppressed the secretion of IL-6 when stimulated by E. coli but not P. gingivalis LPS, implying different signaling mechanism mediated by LPS from those organisms. Data are mean values ± SEM of 2-3 experiments. (D) P. gingivalis LPS stimulation of three separate gingival cell lines, as indicated, resulted in a 2.3 – 8.4 fold increase in CCL25 RNA levels compared to non-stimulated (PBS) cells. Gene expression levels were determined for each sample relative to the 18s gene expression of that sample as described in the Materials and methods. Since all P. gingivalis LPS stimulated cultures had greater CCL25 gene expression than non-stimulated cultures, gene expression levels for non-stimulated cultures were assigned a value of 1.0 and gene expression levels of stimulated cultures were expressed as fold increase over non-stimulated cultures.
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    DuPont de Nemours a p gingivalis
    (A) IL-6 and TNFα levels in tissue culture supernatants from unstimulated (no <t>LPS)</t> and P. <t>gingivalis</t> LPS-stimulated cell lines, and (B) unstimulated (no LPS) and E. coli LPS-stimulated cell lines. (C) Anti-TLR4 antibody suppressed the secretion of IL-6 when stimulated by E. coli but not P. gingivalis LPS, implying different signaling mechanism mediated by LPS from those organisms. Data are mean values ± SEM of 2-3 experiments. (D) P. gingivalis LPS stimulation of three separate gingival cell lines, as indicated, resulted in a 2.3 – 8.4 fold increase in CCL25 RNA levels compared to non-stimulated (PBS) cells. Gene expression levels were determined for each sample relative to the 18s gene expression of that sample as described in the Materials and methods. Since all P. gingivalis LPS stimulated cultures had greater CCL25 gene expression than non-stimulated cultures, gene expression levels for non-stimulated cultures were assigned a value of 1.0 and gene expression levels of stimulated cultures were expressed as fold increase over non-stimulated cultures.
    A P Gingivalis, supplied by DuPont de Nemours, used in various techniques. Bioz Stars score: 94/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Difco p gingivalis 381
    Validation of the germfree-rat model of P. gingivalis -induced alveolar bone loss in rats dually infected with P. <t>gingivalis</t> 381 (Pg381) and the S. gordonii carrier strain (Sg Carrier, Pg251). The mean alveolar bone loss (CEJ:ABC) was calculated as the mean difference between the CEJ and the ABC in millimeters per site for each group ± the standard error of the mean ( n = 8). Groups infected with P. gingivalis 381 alone (Pg381) or P. gingivalis and the S. gordonii carrier (Sg Carrier/Pg381) showed significant alveolar bone loss compared to groups colonized with the S. gordonii carrier alone (Sg Carrier) or sham infected (***, P
    P Gingivalis 381, supplied by Difco, used in various techniques. Bioz Stars score: 86/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Malvern Panalytical p gingivalis w50
    PLNC8 αβ binds to P. <t>gingivalis</t> in a dose-dependent manner. Binding of P. gingivalis to PLNC8 αβ and to anti- P.gingivalis antibodies was analyzed by SPR. Both P. gingivalis ATCC 33277 ( a ) and <t>W50</t> ( b ) were found to bind to immobilized PLNC8 αβ (280 nM). The binding was verified by pre-incubating the bacteria with different concentrations of soluble PLNC8 αβ prior to analysis, which resulted in a significantly reduced binding to the immobilized bacteriocins. Pre-incubation of P. gingivalis ATCC 33277 ( c ) and W50 ( d ) with increasing concentrations of soluble PLNC8 αβ prior to analysis reduced the bacterial binding to anti- P. gingivalis antibodies in a dose-dependent manner. Results are presented from three independent experiments. * p
    P Gingivalis W50, supplied by Malvern Panalytical, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC p gingivalis p gingivalis
    PLNC8 αβ binds to P. <t>gingivalis</t> in a dose-dependent manner. Binding of P. gingivalis to PLNC8 αβ and to anti- P.gingivalis antibodies was analyzed by SPR. Both P. gingivalis ATCC 33277 ( a ) and <t>W50</t> ( b ) were found to bind to immobilized PLNC8 αβ (280 nM). The binding was verified by pre-incubating the bacteria with different concentrations of soluble PLNC8 αβ prior to analysis, which resulted in a significantly reduced binding to the immobilized bacteriocins. Pre-incubation of P. gingivalis ATCC 33277 ( c ) and W50 ( d ) with increasing concentrations of soluble PLNC8 αβ prior to analysis reduced the bacterial binding to anti- P. gingivalis antibodies in a dose-dependent manner. Results are presented from three independent experiments. * p
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    90
    TaKaRa p gingivalis 33277
    Biofilm formation by homotypic P. gingivalis 33277, ECF sigma factor mutant and complemented mutant strain.  The strains were grown on enriched BHI broth anaerobically at 37°C in collagen type-I-coated 12-well flat-bottom microplates. After 48 h of cultivation, the organized biofilm mass was evaluated by staining with crystal violet.  (a)  Biofilm formation of 33277, PGN_0274 mutant and the complemented mutant strain were compared.  (b)  Biofilm formation of 33277, PGN_1740 mutant and the complemented mutant strain were compared. Biofilm formation determined by crystal violet staining and adjusted for growth (A 600  units per OD 660  unit). The data shown are mean ± SD of triplicate experiments. *,  p
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    Image Search Results


    Survival curve of G. mellonella infected with P. gingivalis at different concentrations of pomegranate extract. The concentrations of PGE (12.5 mg/mL, 6.25 mg/mL, 3.1 mg/mL, and 2.5 mg/mL) diluted in PBS were inoculated in the last proleg of each larva. Fifteen larvae were used per group. The number of dead G. mellonella was daily recorded for 168 hours for analysis of the survival curve. The larvae survived at concentrations of 12.5 mg/mL, 6.25 mg/mL, 3.1 mg/mL, and 2.5 mg/mL.

    Journal: The Scientific World Journal

    Article Title: Punica granatum L. (Pomegranate) Extract: In Vivo Study of Antimicrobial Activity against Porphyromonas gingivalis in Galleria mellonella Model

    doi: 10.1155/2016/8626987

    Figure Lengend Snippet: Survival curve of G. mellonella infected with P. gingivalis at different concentrations of pomegranate extract. The concentrations of PGE (12.5 mg/mL, 6.25 mg/mL, 3.1 mg/mL, and 2.5 mg/mL) diluted in PBS were inoculated in the last proleg of each larva. Fifteen larvae were used per group. The number of dead G. mellonella was daily recorded for 168 hours for analysis of the survival curve. The larvae survived at concentrations of 12.5 mg/mL, 6.25 mg/mL, 3.1 mg/mL, and 2.5 mg/mL.

    Article Snippet: Microorganisms and Culture A type strain of P. gingivalis (ATCC 33277) was used.

    Techniques: Infection

    Viability of intracellular WT P. gingivalis (ATCC 33277 and 381) and KDP136 in HAECs. HAECs (1 × 10 5 cells per well) were infected with P. gingivalis ATCC 33277 (black bars), 381 (hatched bars), or KDP136 (white bars) for 20 min at an MOI of 10

    Journal: Infection and Immunity

    Article Title: Role for Gingipains in Porphyromonas gingivalis Traffic to Phagolysosomes and Survival in Human Aortic Endothelial Cells ▿

    doi: 10.1128/IAI.01013-06

    Figure Lengend Snippet: Viability of intracellular WT P. gingivalis (ATCC 33277 and 381) and KDP136 in HAECs. HAECs (1 × 10 5 cells per well) were infected with P. gingivalis ATCC 33277 (black bars), 381 (hatched bars), or KDP136 (white bars) for 20 min at an MOI of 10

    Article Snippet: The HAECs were infected with WT P. gingivalis (strains ATCC 33277, 381 and W83) or KDP136 in kanamycin- and FBS-free MCDB131 medium at an MOI of 103 bacterial cells for 20 min.

    Techniques: Infection

    The J5-c5 transposon mutant induces a proinflammatory response. The amounts of IL-6 secreted by MM6 cells in response to exposure to 10 6 or 10 7 intact P. gingivalis 33277 bacteria versus the isogenic ΔPG1587, ΔPG1773, or Δ kgp mutant (A) and the J5-c5 transposon mutant (B and C) are shown. The results are means ± standard deviations (SD) for triplicate samples from either one of two independent experiments (A) or from two separate experiments (B and C). Asterisks denote significant differences in amount of IL-6 secreted relative to that for the wild-type 33277 control ( P

    Journal: Journal of Bacteriology

    Article Title: Identification of PGN_1123 as the Gene Encoding Lipid A Deacylase, an Enzyme Required for Toll-Like Receptor 4 Evasion, in Porphyromonas gingivalis

    doi: 10.1128/JB.00683-18

    Figure Lengend Snippet: The J5-c5 transposon mutant induces a proinflammatory response. The amounts of IL-6 secreted by MM6 cells in response to exposure to 10 6 or 10 7 intact P. gingivalis 33277 bacteria versus the isogenic ΔPG1587, ΔPG1773, or Δ kgp mutant (A) and the J5-c5 transposon mutant (B and C) are shown. The results are means ± standard deviations (SD) for triplicate samples from either one of two independent experiments (A) or from two separate experiments (B and C). Asterisks denote significant differences in amount of IL-6 secreted relative to that for the wild-type 33277 control ( P

    Article Snippet: P. gingivalis 33277, Bacteroides thetaiotaomicron ATCC 29148, and E. coli DH10b were obtained from our culture collection.

    Techniques: Mutagenesis

    LL-37 is cleaved by Arg-gingipains. Western immunodetection of LL-37 incubated overnight with conditioned medium prepared from P. gingivalis strain KDP129 (ATCC 33277 with kgp inactivated, duplicate in lanes A and B), KDP133 (ATCC 33277 with both rgpA

    Journal: Infection and Immunity

    Article Title: Saliva Enables the Antimicrobial Activity of LL-37 in the Presence of Proteases of Porphyromonas gingivalis ▿ ▿ †

    doi: 10.1128/IAI.00648-09

    Figure Lengend Snippet: LL-37 is cleaved by Arg-gingipains. Western immunodetection of LL-37 incubated overnight with conditioned medium prepared from P. gingivalis strain KDP129 (ATCC 33277 with kgp inactivated, duplicate in lanes A and B), KDP133 (ATCC 33277 with both rgpA

    Article Snippet: P. gingivalis KDP133 ( P. gingivalis ATCC 33277 with rgpA and rgpB inactivated) and P. gingivalis KDP129 ( P. gingivalis ATCC 33277 with kgp inactivated) were generously supplied by Koji Nakayama ( ) and grown in enriched brain heart infusion (BHI) broth (containing 37 g of BHI [Difco, MD], 5 g of yeast extract [Difco], 1 g of cysteine, 5 mg of hemin, and 1 mg of vitamin K1 per liter) supplemented with erythromycin at 10 μg/ml and tetracycline at 0.7 μg/ml (for P. gingivalis KDP133) or with chloramphenicol at 20 μg/ml (for P. gingivalis KDP129).

    Techniques: Western Blot, Immunodetection, Incubation

    Alteration of the transcriptional landscape of TIGKs infected with P. gingivalis in the presence of absence of active PPAD. Significant changes in GS, e.g ., biological processes and pathways, induced in TIGKs upon infection with PPAD-competent P. gingivalis were mapped. The fill color of each node represents the direction and significance level for this comparison (orange, enrichment, FDR q

    Journal: Scientific Reports

    Article Title: Impact of Porphyromonas gingivalis Peptidylarginine Deiminase on Bacterial Biofilm Formation, Epithelial Cell Invasion, and Epithelial Cell Transcriptional Landscape

    doi: 10.1038/s41598-018-32603-y

    Figure Lengend Snippet: Alteration of the transcriptional landscape of TIGKs infected with P. gingivalis in the presence of absence of active PPAD. Significant changes in GS, e.g ., biological processes and pathways, induced in TIGKs upon infection with PPAD-competent P. gingivalis were mapped. The fill color of each node represents the direction and significance level for this comparison (orange, enrichment, FDR q

    Article Snippet: RNA-Seq and mapping of the TIGK transcriptional landscape in response to PPAD activity Global gene expression profiles were generated from the following samples using HumanHT-12 v4 Expression BeadChip kits (Illumina, San Diego, USA): (i) uninfected TIGKs; (ii) TIGKs infected with P. gingivalis WT (ATCC 33277); and (iii) TIGKS infected with P. gingivalis PPADC351A .

    Techniques: Infection

    Identification and functional annotation of TIGK genes significantly dependent on PPAD activity. ( A ) Scatterplot of log2 fold-change (log2FC) in gene expression of TIGKs infected with P. gingivalis WT vs. control (y-axis), and P. gingivalis PPAD C351A vs. P. gingivalis WT (x-axis). Genes altered exclusively in the P. gingivalis WT vs. control comparison are displayed in red when significantly up-regulated (log2FC > 0.5, FDR q

    Journal: Scientific Reports

    Article Title: Impact of Porphyromonas gingivalis Peptidylarginine Deiminase on Bacterial Biofilm Formation, Epithelial Cell Invasion, and Epithelial Cell Transcriptional Landscape

    doi: 10.1038/s41598-018-32603-y

    Figure Lengend Snippet: Identification and functional annotation of TIGK genes significantly dependent on PPAD activity. ( A ) Scatterplot of log2 fold-change (log2FC) in gene expression of TIGKs infected with P. gingivalis WT vs. control (y-axis), and P. gingivalis PPAD C351A vs. P. gingivalis WT (x-axis). Genes altered exclusively in the P. gingivalis WT vs. control comparison are displayed in red when significantly up-regulated (log2FC > 0.5, FDR q

    Article Snippet: RNA-Seq and mapping of the TIGK transcriptional landscape in response to PPAD activity Global gene expression profiles were generated from the following samples using HumanHT-12 v4 Expression BeadChip kits (Illumina, San Diego, USA): (i) uninfected TIGKs; (ii) TIGKs infected with P. gingivalis WT (ATCC 33277); and (iii) TIGKS infected with P. gingivalis PPADC351A .

    Techniques: Functional Assay, Activity Assay, Expressing, Infection

    The effect of P. gingivalis PPAD on bacterial abundance ( a ), species composition ( b ) and ( c ) the biofilm structure in multispecies biofilms. A biofilm consisting of T. forsythia , F. nucleatum , A. naeslundii , S. gordonii , and one of three P. gingivalis strains, wild type (WT), ppad deletion strain ( Δppad ), or a strain harboring inactivated PPAD (C351A), was cultured for 48 h, following which ( a,b ) the bacterial DNA was extracted and assessed via qPCR, ( c ) biofilm was fixed and observed via SEM under 1500x magnification. The ( a ) bacterial load and ( b ) species composition of each biofilm model are presented as mean ± SD from three independent experiments. Data were plotted on a logarithmic scale.

    Journal: Scientific Reports

    Article Title: Impact of Porphyromonas gingivalis Peptidylarginine Deiminase on Bacterial Biofilm Formation, Epithelial Cell Invasion, and Epithelial Cell Transcriptional Landscape

    doi: 10.1038/s41598-018-32603-y

    Figure Lengend Snippet: The effect of P. gingivalis PPAD on bacterial abundance ( a ), species composition ( b ) and ( c ) the biofilm structure in multispecies biofilms. A biofilm consisting of T. forsythia , F. nucleatum , A. naeslundii , S. gordonii , and one of three P. gingivalis strains, wild type (WT), ppad deletion strain ( Δppad ), or a strain harboring inactivated PPAD (C351A), was cultured for 48 h, following which ( a,b ) the bacterial DNA was extracted and assessed via qPCR, ( c ) biofilm was fixed and observed via SEM under 1500x magnification. The ( a ) bacterial load and ( b ) species composition of each biofilm model are presented as mean ± SD from three independent experiments. Data were plotted on a logarithmic scale.

    Article Snippet: RNA-Seq and mapping of the TIGK transcriptional landscape in response to PPAD activity Global gene expression profiles were generated from the following samples using HumanHT-12 v4 Expression BeadChip kits (Illumina, San Diego, USA): (i) uninfected TIGKs; (ii) TIGKs infected with P. gingivalis WT (ATCC 33277); and (iii) TIGKS infected with P. gingivalis PPADC351A .

    Techniques: Cell Culture, Real-time Polymerase Chain Reaction

    P. gingivalis invasion of human oral keratinocytes. CTV-labeled TIGKs were infected with various CFSE-labeled strains of P. gingivalis (MOI = 200) for 90 min. Metronidazole was added for an additional 1 h, following which the cells were fixed and analyzed by flow cytometry. Cells were first gated on the basis of a forward scatter/side scatter (FSC-A/SSC-A) plot ( a ). The events were then visualized using FSC-A/FSC-H dot plot, and single cells were gated ( b ). TIGKs were identified on the basis of CTV positivity ( c ). TIGKs invaded by P. gingivalis were subsequently defined as the CFSE + cells within the CTV + keratinocyte population ( d ). Overall, 20,000 events were analyzed, and the results are presented as the percentage of TIGKs infected with P. gingivalis wild-type ( e ), Δppad ( f ), and C351A ( g ).

    Journal: Scientific Reports

    Article Title: Impact of Porphyromonas gingivalis Peptidylarginine Deiminase on Bacterial Biofilm Formation, Epithelial Cell Invasion, and Epithelial Cell Transcriptional Landscape

    doi: 10.1038/s41598-018-32603-y

    Figure Lengend Snippet: P. gingivalis invasion of human oral keratinocytes. CTV-labeled TIGKs were infected with various CFSE-labeled strains of P. gingivalis (MOI = 200) for 90 min. Metronidazole was added for an additional 1 h, following which the cells were fixed and analyzed by flow cytometry. Cells were first gated on the basis of a forward scatter/side scatter (FSC-A/SSC-A) plot ( a ). The events were then visualized using FSC-A/FSC-H dot plot, and single cells were gated ( b ). TIGKs were identified on the basis of CTV positivity ( c ). TIGKs invaded by P. gingivalis were subsequently defined as the CFSE + cells within the CTV + keratinocyte population ( d ). Overall, 20,000 events were analyzed, and the results are presented as the percentage of TIGKs infected with P. gingivalis wild-type ( e ), Δppad ( f ), and C351A ( g ).

    Article Snippet: RNA-Seq and mapping of the TIGK transcriptional landscape in response to PPAD activity Global gene expression profiles were generated from the following samples using HumanHT-12 v4 Expression BeadChip kits (Illumina, San Diego, USA): (i) uninfected TIGKs; (ii) TIGKs infected with P. gingivalis WT (ATCC 33277); and (iii) TIGKS infected with P. gingivalis PPADC351A .

    Techniques: Labeling, Infection, Flow Cytometry, Cytometry

    Fas expression by CD3 + T cells derived from chronic periodontitis (CP) patients and subjects without periodontitis (NP) upon stimulation (48 h) with P. gingivalis ATCC 33277 crude extract (Pg33277), purified recombinant P. gingivalis HmuY protein (HmuY), or without stimulus (Cells) as evaluated by flow cytometry.

    Journal: BMC Microbiology

    Article Title: Porphyromonas gingivalis HmuY stimulates expression of Bcl-2 and Fas by human CD3+ T cells

    doi: 10.1186/1471-2180-13-206

    Figure Lengend Snippet: Fas expression by CD3 + T cells derived from chronic periodontitis (CP) patients and subjects without periodontitis (NP) upon stimulation (48 h) with P. gingivalis ATCC 33277 crude extract (Pg33277), purified recombinant P. gingivalis HmuY protein (HmuY), or without stimulus (Cells) as evaluated by flow cytometry.

    Article Snippet: Cells were also incubated with 5 μg/mL of pokeweed mitogen (PWM) as a positive control, 0.5 μg/mL of P. gingivalis extract (ATCC 33277), 2.5 μg/mL of HmuY, or in the absence of antigens (Cells).

    Techniques: Expressing, Derivative Assay, Purification, Recombinant, Flow Cytometry, Cytometry

    Bcl-2 expression by CD3 + T cells derived from chronic periodontitis (CP) patients and subjects without periodontitis (NP) upon stimulation (48 h) with P. gingivalis ATCC 33277 crude extract (Pg33277), purified recombinant P. gingivalis HmuY protein (HmuY), or without stimulus (Cells) as evaluated by flow cytometry. * p = 0.043, ‡ p = 0,011.

    Journal: BMC Microbiology

    Article Title: Porphyromonas gingivalis HmuY stimulates expression of Bcl-2 and Fas by human CD3+ T cells

    doi: 10.1186/1471-2180-13-206

    Figure Lengend Snippet: Bcl-2 expression by CD3 + T cells derived from chronic periodontitis (CP) patients and subjects without periodontitis (NP) upon stimulation (48 h) with P. gingivalis ATCC 33277 crude extract (Pg33277), purified recombinant P. gingivalis HmuY protein (HmuY), or without stimulus (Cells) as evaluated by flow cytometry. * p = 0.043, ‡ p = 0,011.

    Article Snippet: Cells were also incubated with 5 μg/mL of pokeweed mitogen (PWM) as a positive control, 0.5 μg/mL of P. gingivalis extract (ATCC 33277), 2.5 μg/mL of HmuY, or in the absence of antigens (Cells).

    Techniques: Expressing, Derivative Assay, Purification, Recombinant, Flow Cytometry, Cytometry

    Binding of Alexa Fluor-labeled LL-37 to E. coli ATCC 25922 or P. gingivalis KDP136. (A) Total fluorescence of bacteria incubated with Alexa Fluor-labeled LL-37 (concentration is estimated at 0.8 mg/ml [see Materials and Methods]). (B) Total fluorescence

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Resistance of Porphyromonas gingivalis ATCC 33277 to Direct Killing by Antimicrobial Peptides Is Protease Independent ▿

    doi: 10.1128/AAC.01271-07

    Figure Lengend Snippet: Binding of Alexa Fluor-labeled LL-37 to E. coli ATCC 25922 or P. gingivalis KDP136. (A) Total fluorescence of bacteria incubated with Alexa Fluor-labeled LL-37 (concentration is estimated at 0.8 mg/ml [see Materials and Methods]). (B) Total fluorescence

    Article Snippet: P. gingivalis KDP136 ( P. gingivalis ATCC 33277 mutant inactivated in rgpA , rgpB , and kgp ), generously supplied by Koji Nakayama , was grown in enriched BHI broth (containing, per liter, 37 g of BHI [Difco], 5 g of yeast extract [Difco], 1 g of cysteine, 5 mg of hemin, and 1 mg of vitamin K1 ) supplemented with chloramphenicol, 20 μg/ml; erythromycin, 10 μg/ml; and tetracycline, 0.7 μg/ml.

    Techniques: Binding Assay, Labeling, Fluorescence, Incubation, Concentration Assay

    Adoptive transfer of CD1d hi CD5 + B cells inhibits P. gingivalis- associated ligature-induced periodontal bone loss. P. gingivalis -soaked ligatures were placed subgingivally around the maxillary second molars of C57BL/6 mice on day 1 and were retained for 2 weeks to establish an experimental periodontitis model. CD1d hi CD5 + B cells and CD1d lo CD5 − B cells were sorted from splenic B cells separated from P. gingivalis- immunized donor mice by flow cytometry and then transferred (1 × 10 6 cells in 100 μl PBS per mouse) into recipient mice through the tail vein on day 2; the same amount of PBS were injected into the control group without ligation and the group with ligation and no transfer. (a) Maxillae were collected on day 14, and the buccal and palatal alveolar bone resorption areas around maxillary second molars were measured. (b) Data are presented as bone resorption area per square millimeter at a magnification of ×30 (means ± standard errors; n = 6) (**, P

    Journal: Infection and Immunity

    Article Title: B10 Cells Alleviate Periodontal Bone Loss in Experimental Periodontitis

    doi: 10.1128/IAI.00335-17

    Figure Lengend Snippet: Adoptive transfer of CD1d hi CD5 + B cells inhibits P. gingivalis- associated ligature-induced periodontal bone loss. P. gingivalis -soaked ligatures were placed subgingivally around the maxillary second molars of C57BL/6 mice on day 1 and were retained for 2 weeks to establish an experimental periodontitis model. CD1d hi CD5 + B cells and CD1d lo CD5 − B cells were sorted from splenic B cells separated from P. gingivalis- immunized donor mice by flow cytometry and then transferred (1 × 10 6 cells in 100 μl PBS per mouse) into recipient mice through the tail vein on day 2; the same amount of PBS were injected into the control group without ligation and the group with ligation and no transfer. (a) Maxillae were collected on day 14, and the buccal and palatal alveolar bone resorption areas around maxillary second molars were measured. (b) Data are presented as bone resorption area per square millimeter at a magnification of ×30 (means ± standard errors; n = 6) (**, P

    Article Snippet: P. gingivalis bacteria (strain ATCC 33277) were grown on anaerobic blood agar plates (NHK agar; Northeast Laboratory Services, Waterville, ME) in an anaerobic chamber with 85% N2 , 5% H2 , and 10% CO2 .

    Techniques: Adoptive Transfer Assay, Mouse Assay, Flow Cytometry, Cytometry, Injection, Ligation

    Changes in IL-10 mRNA and protein levels in mouse splenocyte B cells after LPS, CpG, and LPS+CpG treatment. Splenocyte B cells were separated from nonimmunized C57BL/6J mice and cultured with P. gingivalis LPS (10 μg/ml), CpG (10 μM), and P. gingivalis LPS (10 μg/ml) plus CpG (10 μM) for 48 h. (a) IL-10 mRNA expression levels in cell lysates of the control, LPS, CpG, and LPS+CpG groups were determined by real-time PCR in duplicates. (b) Secreted IL-10 protein levels in the supernatants of the same groups as the ones mentioned above were measured in duplicates by using an enzyme-linked immunosorbent assay kit (means ± standard errors; n = 6) (*, P

    Journal: Infection and Immunity

    Article Title: B10 Cells Alleviate Periodontal Bone Loss in Experimental Periodontitis

    doi: 10.1128/IAI.00335-17

    Figure Lengend Snippet: Changes in IL-10 mRNA and protein levels in mouse splenocyte B cells after LPS, CpG, and LPS+CpG treatment. Splenocyte B cells were separated from nonimmunized C57BL/6J mice and cultured with P. gingivalis LPS (10 μg/ml), CpG (10 μM), and P. gingivalis LPS (10 μg/ml) plus CpG (10 μM) for 48 h. (a) IL-10 mRNA expression levels in cell lysates of the control, LPS, CpG, and LPS+CpG groups were determined by real-time PCR in duplicates. (b) Secreted IL-10 protein levels in the supernatants of the same groups as the ones mentioned above were measured in duplicates by using an enzyme-linked immunosorbent assay kit (means ± standard errors; n = 6) (*, P

    Article Snippet: P. gingivalis bacteria (strain ATCC 33277) were grown on anaerobic blood agar plates (NHK agar; Northeast Laboratory Services, Waterville, ME) in an anaerobic chamber with 85% N2 , 5% H2 , and 10% CO2 .

    Techniques: Mouse Assay, Cell Culture, Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

    B10 cell expansion in mouse splenocyte B cells after LPS, CpG, and LPS+CpG treatments and IL-10 mRNA expression levels in the CD1d hi CD5 + B cell subset. Splenocyte B cells were separated from nonimmunized C57BL/6J mice and cultured with P. gingivalis LPS (10 μg/ml), CpG (10 μM), and P. gingivalis LPS (10 μg/ml) plus CpG (10 μM) for 48 h. (a) IL-10-expressing B cells (CD19 + IL-10 + B cells) in control and treatment groups were detected by using flow cytometry in duplicates. (b) The percentages of CD19 + IL-10 + B cells in control and treatment groups were quantified and analyzed by using FlowJo software (means ± standard deviations; n = 5) (*, P

    Journal: Infection and Immunity

    Article Title: B10 Cells Alleviate Periodontal Bone Loss in Experimental Periodontitis

    doi: 10.1128/IAI.00335-17

    Figure Lengend Snippet: B10 cell expansion in mouse splenocyte B cells after LPS, CpG, and LPS+CpG treatments and IL-10 mRNA expression levels in the CD1d hi CD5 + B cell subset. Splenocyte B cells were separated from nonimmunized C57BL/6J mice and cultured with P. gingivalis LPS (10 μg/ml), CpG (10 μM), and P. gingivalis LPS (10 μg/ml) plus CpG (10 μM) for 48 h. (a) IL-10-expressing B cells (CD19 + IL-10 + B cells) in control and treatment groups were detected by using flow cytometry in duplicates. (b) The percentages of CD19 + IL-10 + B cells in control and treatment groups were quantified and analyzed by using FlowJo software (means ± standard deviations; n = 5) (*, P

    Article Snippet: P. gingivalis bacteria (strain ATCC 33277) were grown on anaerobic blood agar plates (NHK agar; Northeast Laboratory Services, Waterville, ME) in an anaerobic chamber with 85% N2 , 5% H2 , and 10% CO2 .

    Techniques: Expressing, Mouse Assay, Cell Culture, Flow Cytometry, Cytometry, Software

    Adoptive transfer of CD1d hi CD5 + B cells increases the numbers of IL-10-expressing B cells, CD1d hi CD5 + B cells, and P. gingivalis -binding B cells in gingival tissue. Gingival tissues from mice of the no-transfer control group, the CD1d lo CD5 − B cell transfer group, and the CD1d hi CD5 + B cell transfer group were isolated under a surgical microscope at day 14 after ligation. Mononuclear cells were recovered from gingival tissues of each group (total number of cells ranging from 1.3 × 10 2 to 2.7 × 10 3 cells per tissue). The percentages of CD19 + IL-10 + cells (a and d), CD19 + CD1d hi CD5 + cells (b and e), and P. gingivalis -binding CD19 + cells (c and f) in these mononuclear cells were measured by flow cytometry in duplicates and were analyzed by using FlowJo software (means ± standard errors; n = 6) (**, P

    Journal: Infection and Immunity

    Article Title: B10 Cells Alleviate Periodontal Bone Loss in Experimental Periodontitis

    doi: 10.1128/IAI.00335-17

    Figure Lengend Snippet: Adoptive transfer of CD1d hi CD5 + B cells increases the numbers of IL-10-expressing B cells, CD1d hi CD5 + B cells, and P. gingivalis -binding B cells in gingival tissue. Gingival tissues from mice of the no-transfer control group, the CD1d lo CD5 − B cell transfer group, and the CD1d hi CD5 + B cell transfer group were isolated under a surgical microscope at day 14 after ligation. Mononuclear cells were recovered from gingival tissues of each group (total number of cells ranging from 1.3 × 10 2 to 2.7 × 10 3 cells per tissue). The percentages of CD19 + IL-10 + cells (a and d), CD19 + CD1d hi CD5 + cells (b and e), and P. gingivalis -binding CD19 + cells (c and f) in these mononuclear cells were measured by flow cytometry in duplicates and were analyzed by using FlowJo software (means ± standard errors; n = 6) (**, P

    Article Snippet: P. gingivalis bacteria (strain ATCC 33277) were grown on anaerobic blood agar plates (NHK agar; Northeast Laboratory Services, Waterville, ME) in an anaerobic chamber with 85% N2 , 5% H2 , and 10% CO2 .

    Techniques: Adoptive Transfer Assay, Expressing, Binding Assay, Mouse Assay, Isolation, Microscopy, Ligation, Flow Cytometry, Cytometry, Software

    PPAD supplementation restores the ability of P. gingivalis ΔPPAD to adhere to and invade primary human gingival fibroblasts (PHGF)

    Journal: Molecular oral microbiology

    Article Title: Peptidylarginine deiminase from Porphyromonas gingivalis (PPAD) contributes to infection of gingival fibroblasts and induction of PGE2-signaling pathway

    doi: 10.1111/omi.12081

    Figure Lengend Snippet: PPAD supplementation restores the ability of P. gingivalis ΔPPAD to adhere to and invade primary human gingival fibroblasts (PHGF)

    Article Snippet: P. gingivalis strains (wt- Pg ATCC 33277 and its isogenic mutant Δ ppad and C351A expressing catalytically inactive PPAD (PPADC351A ) were grown on blood agar plates (BHI, brain heart infusion medium supplemented with 5% sheep blood, 5 μg mL−1 hemin and 0.5 μg mL−1 vitamin K in an anaerobic chamber (90% N2 , 10% CO2 , and 5% H2 ).

    Techniques:

    P. gingivalis adheres to and invades primary human gingival fibroblasts (PHGF) more efficiently than P. gingivalis- ΔPPAD mutants

    Journal: Molecular oral microbiology

    Article Title: Peptidylarginine deiminase from Porphyromonas gingivalis (PPAD) contributes to infection of gingival fibroblasts and induction of PGE2-signaling pathway

    doi: 10.1111/omi.12081

    Figure Lengend Snippet: P. gingivalis adheres to and invades primary human gingival fibroblasts (PHGF) more efficiently than P. gingivalis- ΔPPAD mutants

    Article Snippet: P. gingivalis strains (wt- Pg ATCC 33277 and its isogenic mutant Δ ppad and C351A expressing catalytically inactive PPAD (PPADC351A ) were grown on blood agar plates (BHI, brain heart infusion medium supplemented with 5% sheep blood, 5 μg mL−1 hemin and 0.5 μg mL−1 vitamin K in an anaerobic chamber (90% N2 , 10% CO2 , and 5% H2 ).

    Techniques:

    PPAD is involved in efficient interaction with, adhesion to and invasion of gingival fibroblasts by P. gingivalis

    Journal: Molecular oral microbiology

    Article Title: Peptidylarginine deiminase from Porphyromonas gingivalis (PPAD) contributes to infection of gingival fibroblasts and induction of PGE2-signaling pathway

    doi: 10.1111/omi.12081

    Figure Lengend Snippet: PPAD is involved in efficient interaction with, adhesion to and invasion of gingival fibroblasts by P. gingivalis

    Article Snippet: P. gingivalis strains (wt- Pg ATCC 33277 and its isogenic mutant Δ ppad and C351A expressing catalytically inactive PPAD (PPADC351A ) were grown on blood agar plates (BHI, brain heart infusion medium supplemented with 5% sheep blood, 5 μg mL−1 hemin and 0.5 μg mL−1 vitamin K in an anaerobic chamber (90% N2 , 10% CO2 , and 5% H2 ).

    Techniques:

    Gingipain deficiency reverses the suppression of F. nucleatum invasion by P. gingivalis . HOK-16B cells were infected with CFSE-labeled F. nucleatum and unlabeled wild-type P. gingivalis or gingipain mutants at a MOI of 500 for 4 h. (A) Cells were analyzed using flow cytometry after quenching the extracellular fluorescence from the bacteria attached to the cell surface with trypan blue. The percentage of cells containing F. nucleatum (left panel) and the MFI (right panel) are shown as the mean ± standard deviation. Representative data from three independent experiments are shown. (B) The cells were examined under a confocal laser scanning microscope after staining for F-actin (red) and nuclei (blue). CFSE-labeled F. nucleatum is displayed in green. Fn, F. nucleatum ; Pg, P. gingivalis ; WT, P. gingivalis ATCC 33277; 129, KDP129 ( kgp − ); 133, KDP133 ( rgpA − rgpB − ); 136, KDP136 ( kgp − rgpA − rgpB − ). * p

    Journal: Journal of Oral Microbiology

    Article Title: Porphyromonas gingivalis suppresses invasion of Fusobacterium nucleatum into gingival epithelial cells

    doi: 10.1080/20002297.2017.1320193

    Figure Lengend Snippet: Gingipain deficiency reverses the suppression of F. nucleatum invasion by P. gingivalis . HOK-16B cells were infected with CFSE-labeled F. nucleatum and unlabeled wild-type P. gingivalis or gingipain mutants at a MOI of 500 for 4 h. (A) Cells were analyzed using flow cytometry after quenching the extracellular fluorescence from the bacteria attached to the cell surface with trypan blue. The percentage of cells containing F. nucleatum (left panel) and the MFI (right panel) are shown as the mean ± standard deviation. Representative data from three independent experiments are shown. (B) The cells were examined under a confocal laser scanning microscope after staining for F-actin (red) and nuclei (blue). CFSE-labeled F. nucleatum is displayed in green. Fn, F. nucleatum ; Pg, P. gingivalis ; WT, P. gingivalis ATCC 33277; 129, KDP129 ( kgp − ); 133, KDP133 ( rgpA − rgpB − ); 136, KDP136 ( kgp − rgpA − rgpB − ). * p

    Article Snippet: Wild-type P. gingivalis (ATCC 33277) and the mutant strains were grown anaerobically at 37°C in enriched BHI broth supplemented with 5 μg/mL of hemin and 1 μg/mL of vitamin K, and the following antibiotics, as described previously [ ]: chloramphenicol (20 μg/mL), erythromycin (10 μg/mL), and tetracycline (0.7 μg/mL).

    Techniques: Infection, Labeling, Flow Cytometry, Cytometry, Fluorescence, Standard Deviation, Laser-Scanning Microscopy, Staining

    P . gingivalis inactivates the PI3K/AKT pathway in a gingipain-dependent manner. (A) HOK-16B cells were infected with wild-type P. gingivalis at a MOI of 500 for the indicated time. (B) and (C) HOK-16B cells were infected with wild-type P. gingivalis or gingipain mutants in the presence or absence of F. nucleatum at a MOI of 500 for 30 min (B) or 3 h (C). The levels of phosphorylated and total forms of PI3K and AKT were analyzed by immunoblot analysis. Fn, F. nucleatum ; Pg, P. gingivalis ; WT, P. gingivalis ATCC 33277; 129, KDP129 ( kgp − ); 133, KDP133 ( rgpA − rgpB − ); 136, KDP136 ( kgp − rgpA − rgpB − ).

    Journal: Journal of Oral Microbiology

    Article Title: Porphyromonas gingivalis suppresses invasion of Fusobacterium nucleatum into gingival epithelial cells

    doi: 10.1080/20002297.2017.1320193

    Figure Lengend Snippet: P . gingivalis inactivates the PI3K/AKT pathway in a gingipain-dependent manner. (A) HOK-16B cells were infected with wild-type P. gingivalis at a MOI of 500 for the indicated time. (B) and (C) HOK-16B cells were infected with wild-type P. gingivalis or gingipain mutants in the presence or absence of F. nucleatum at a MOI of 500 for 30 min (B) or 3 h (C). The levels of phosphorylated and total forms of PI3K and AKT were analyzed by immunoblot analysis. Fn, F. nucleatum ; Pg, P. gingivalis ; WT, P. gingivalis ATCC 33277; 129, KDP129 ( kgp − ); 133, KDP133 ( rgpA − rgpB − ); 136, KDP136 ( kgp − rgpA − rgpB − ).

    Article Snippet: Wild-type P. gingivalis (ATCC 33277) and the mutant strains were grown anaerobically at 37°C in enriched BHI broth supplemented with 5 μg/mL of hemin and 1 μg/mL of vitamin K, and the following antibiotics, as described previously [ ]: chloramphenicol (20 μg/mL), erythromycin (10 μg/mL), and tetracycline (0.7 μg/mL).

    Techniques: Infection

    Invasion of F. nucleatum is suppressed by PI3K inhibition. (A) HOK-16B cells were preincubated with wortmannin at the indicated concentration for 30 min and then infected with CFSE-labeled F. nucleatum and unlabeled wild-type P. gingivalis at a MOI of 500 for 4 h. The percentage of cells containing F. nucleatum (left panel) and the MFI (right panel) are shown as the mean ± standard deviation. * p

    Journal: Journal of Oral Microbiology

    Article Title: Porphyromonas gingivalis suppresses invasion of Fusobacterium nucleatum into gingival epithelial cells

    doi: 10.1080/20002297.2017.1320193

    Figure Lengend Snippet: Invasion of F. nucleatum is suppressed by PI3K inhibition. (A) HOK-16B cells were preincubated with wortmannin at the indicated concentration for 30 min and then infected with CFSE-labeled F. nucleatum and unlabeled wild-type P. gingivalis at a MOI of 500 for 4 h. The percentage of cells containing F. nucleatum (left panel) and the MFI (right panel) are shown as the mean ± standard deviation. * p

    Article Snippet: Wild-type P. gingivalis (ATCC 33277) and the mutant strains were grown anaerobically at 37°C in enriched BHI broth supplemented with 5 μg/mL of hemin and 1 μg/mL of vitamin K, and the following antibiotics, as described previously [ ]: chloramphenicol (20 μg/mL), erythromycin (10 μg/mL), and tetracycline (0.7 μg/mL).

    Techniques: Inhibition, Concentration Assay, Infection, Labeling, Standard Deviation

    More intracellular F. nucleatum remains viable after coinfection with P. gingivalis with Rgp mutation than in monoinfection. After HOK-16B cells were infected with F. nucleatum in the presence or absence of wild-type P. gingivalis or gingipain mutants at a MOI of 500 for 4 h, extracellular bacteria were killed by incubation with antibiotics for 1 h. After incubation in fresh media for 12 h, the cells were lysed, and the lysates were plated on brain heart infusion blood agar plates containing vancomycin. The data represent the mean ± standard deviation of four independent experiments. * p

    Journal: Journal of Oral Microbiology

    Article Title: Porphyromonas gingivalis suppresses invasion of Fusobacterium nucleatum into gingival epithelial cells

    doi: 10.1080/20002297.2017.1320193

    Figure Lengend Snippet: More intracellular F. nucleatum remains viable after coinfection with P. gingivalis with Rgp mutation than in monoinfection. After HOK-16B cells were infected with F. nucleatum in the presence or absence of wild-type P. gingivalis or gingipain mutants at a MOI of 500 for 4 h, extracellular bacteria were killed by incubation with antibiotics for 1 h. After incubation in fresh media for 12 h, the cells were lysed, and the lysates were plated on brain heart infusion blood agar plates containing vancomycin. The data represent the mean ± standard deviation of four independent experiments. * p

    Article Snippet: Wild-type P. gingivalis (ATCC 33277) and the mutant strains were grown anaerobically at 37°C in enriched BHI broth supplemented with 5 μg/mL of hemin and 1 μg/mL of vitamin K, and the following antibiotics, as described previously [ ]: chloramphenicol (20 μg/mL), erythromycin (10 μg/mL), and tetracycline (0.7 μg/mL).

    Techniques: Mutagenesis, Infection, Incubation, Standard Deviation

    Expression of bacterial glutaminyl cyclase (A: mRNA; B: protein) in P. gingivalis ATCC 33277 and P. gingivalis J430–1 cultures after anaerobic incubation for 1 h, 2 h, 4 h and 24 h

    Journal: Archives of oral biology

    Article Title: Expression of human and Porphyromonas gingivalis glutaminyl cyclases in periodontitis and rheumatoid arthritis - a pilot study

    doi: 10.1016/j.archoralbio.2018.10.022

    Figure Lengend Snippet: Expression of bacterial glutaminyl cyclase (A: mRNA; B: protein) in P. gingivalis ATCC 33277 and P. gingivalis J430–1 cultures after anaerobic incubation for 1 h, 2 h, 4 h and 24 h

    Article Snippet: In preliminary experiments P. gingivalis ATCC 33277 (the reference strain) and P. gingivalis J430–1 (clinical isolate) were included.

    Techniques: Expressing, Incubation

    The effect of AEA on the production of pro-inflammatory mediators in hPdLCs in response to stimulation with P. gingivalis LPS. hPdLC were stimulated with P. gingivalis LPS in the presence or in the absence of AEA for 24 h, and the production of pro-inflammatory mediators was measured on gene and protein levels by real-time PCR (A) and ELISA (B) respectively. A – Changes in the gene expression levels of IL-6, IL-8, and MCP-1 calculated by 2 −ΔΔCt method taking non-stimulated cells as a reference (2 −ΔΔCt = 1) and β-actin as a house-keeping gene. B – Content of pro-inflammatory mediators in the conditioned media measured by ELISA. Each value represents the mean ± s.e.m of 4 independent experiments. # Means were significantly compared to the control group, and tested with analysis of variance (P

    Journal: PLoS ONE

    Article Title: Endocannabinoids and Inflammatory Response in Periodontal Ligament Cells

    doi: 10.1371/journal.pone.0107407

    Figure Lengend Snippet: The effect of AEA on the production of pro-inflammatory mediators in hPdLCs in response to stimulation with P. gingivalis LPS. hPdLC were stimulated with P. gingivalis LPS in the presence or in the absence of AEA for 24 h, and the production of pro-inflammatory mediators was measured on gene and protein levels by real-time PCR (A) and ELISA (B) respectively. A – Changes in the gene expression levels of IL-6, IL-8, and MCP-1 calculated by 2 −ΔΔCt method taking non-stimulated cells as a reference (2 −ΔΔCt = 1) and β-actin as a house-keeping gene. B – Content of pro-inflammatory mediators in the conditioned media measured by ELISA. Each value represents the mean ± s.e.m of 4 independent experiments. # Means were significantly compared to the control group, and tested with analysis of variance (P

    Article Snippet: The gene-expression levels of MCP-1 after stimulation with P. gingivalis were significantly increased in the presence of 10 µM 2-AG but no significant differences was observed on protein level.

    Techniques: Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Expressing

    The effect of 2-AG on the production of pro-inflammatory mediators in hPdLCs in response to stimulation with P. gingivalis LPS. hPdLC were stimulated with P. gingivalis LPS in the presence or in the absence of 2-AG for 24 h and the production of pro-inflammatory mediators was measured on gene and protein levels by real-time PCR (A) and ELISA (B) respectively. A – Changes in the gene expression levels of IL-6, IL-8, and MCP-1 calculated by 2 −ΔΔCt method taking non-stimulated cells as a reference (2 −ΔΔCt = 1) and β-actin as a house-keeping gene. B – Content of pro-inflammatory mediators in the conditioned media measured by ELISA. Each value represents the mean ± s.e.m of 4 independent experiments. # Means were significantly compared to control group, tested with analysis of variance (P

    Journal: PLoS ONE

    Article Title: Endocannabinoids and Inflammatory Response in Periodontal Ligament Cells

    doi: 10.1371/journal.pone.0107407

    Figure Lengend Snippet: The effect of 2-AG on the production of pro-inflammatory mediators in hPdLCs in response to stimulation with P. gingivalis LPS. hPdLC were stimulated with P. gingivalis LPS in the presence or in the absence of 2-AG for 24 h and the production of pro-inflammatory mediators was measured on gene and protein levels by real-time PCR (A) and ELISA (B) respectively. A – Changes in the gene expression levels of IL-6, IL-8, and MCP-1 calculated by 2 −ΔΔCt method taking non-stimulated cells as a reference (2 −ΔΔCt = 1) and β-actin as a house-keeping gene. B – Content of pro-inflammatory mediators in the conditioned media measured by ELISA. Each value represents the mean ± s.e.m of 4 independent experiments. # Means were significantly compared to control group, tested with analysis of variance (P

    Article Snippet: The gene-expression levels of MCP-1 after stimulation with P. gingivalis were significantly increased in the presence of 10 µM 2-AG but no significant differences was observed on protein level.

    Techniques: Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Expressing

    The effects of endocannabinoids on proliferation/viability of hPdLCs. hPdLCs were stimulated by different concentrations of AEA (A) or 2-AG (B) in the presence or in the absence of P. gingivalis LPS (1 µg/ml) for 24 h and the proliferation/viability was measured by the MTT method. Cells stimulated with DMEM supplemented by 1% FCS were used as a control (Co). The Y-axis represents mean ± s.e.m. of optical densities measured at 570 nm in 4 independent experiments. *Means were significantly different between groups (P

    Journal: PLoS ONE

    Article Title: Endocannabinoids and Inflammatory Response in Periodontal Ligament Cells

    doi: 10.1371/journal.pone.0107407

    Figure Lengend Snippet: The effects of endocannabinoids on proliferation/viability of hPdLCs. hPdLCs were stimulated by different concentrations of AEA (A) or 2-AG (B) in the presence or in the absence of P. gingivalis LPS (1 µg/ml) for 24 h and the proliferation/viability was measured by the MTT method. Cells stimulated with DMEM supplemented by 1% FCS were used as a control (Co). The Y-axis represents mean ± s.e.m. of optical densities measured at 570 nm in 4 independent experiments. *Means were significantly different between groups (P

    Article Snippet: The gene-expression levels of MCP-1 after stimulation with P. gingivalis were significantly increased in the presence of 10 µM 2-AG but no significant differences was observed on protein level.

    Techniques: MTT Assay

    Chemokines and cytokines secreted in HCAEC stimulated with P. gingivalis -LPS or live- P. gingivalis . Monolayers of HCAEC cultured in 12-well plates were stimulated with P. gingivalis -LPS (1.0, 3.5, 7.0 µg/mL) or P. gingivalis (MOI 1:100 - 1:0,1) for 24 h under repeated exposure (+++) or single exposure (+). After stimulation, levels of the following chemokines were measured in cell culture supernatants using a cytometric bead array: ( A ) IL-8, ( B ) MCP-1, ( C ) IL-6, ( D ) GM-CSF, ( E ) IL-1β. Symbol (*) means p

    Journal: Scientific Reports

    Article Title: Repeated Porphyromonas gingivalis W83 exposure leads to release pro-inflammatory cytokynes and angiotensin II in coronary artery endothelial cells

    doi: 10.1038/s41598-019-54259-y

    Figure Lengend Snippet: Chemokines and cytokines secreted in HCAEC stimulated with P. gingivalis -LPS or live- P. gingivalis . Monolayers of HCAEC cultured in 12-well plates were stimulated with P. gingivalis -LPS (1.0, 3.5, 7.0 µg/mL) or P. gingivalis (MOI 1:100 - 1:0,1) for 24 h under repeated exposure (+++) or single exposure (+). After stimulation, levels of the following chemokines were measured in cell culture supernatants using a cytometric bead array: ( A ) IL-8, ( B ) MCP-1, ( C ) IL-6, ( D ) GM-CSF, ( E ) IL-1β. Symbol (*) means p

    Article Snippet: The findings of this study suggest that repeated exposure of P. gingivalis in HCAEC induces the activation of proinflammatory and vasoconstrictor molecules that lead to endothelial dysfunction as a key mechanism of the onset and progression of arterial hypertension (HT) and atherosclerosis, which requires more research.

    Techniques: Cell Culture

    Angiotensin II levels are determined in the HCAEC cell culture supernatant stimulated to single (+) or repetitive (+++) exposures of P. gingivalis -LPS or live- P . gingivalis by the ELISA kit. The results are expressed as the means ± SEM (n=3) with a statistical significance represented as (*)p

    Journal: Scientific Reports

    Article Title: Repeated Porphyromonas gingivalis W83 exposure leads to release pro-inflammatory cytokynes and angiotensin II in coronary artery endothelial cells

    doi: 10.1038/s41598-019-54259-y

    Figure Lengend Snippet: Angiotensin II levels are determined in the HCAEC cell culture supernatant stimulated to single (+) or repetitive (+++) exposures of P. gingivalis -LPS or live- P . gingivalis by the ELISA kit. The results are expressed as the means ± SEM (n=3) with a statistical significance represented as (*)p

    Article Snippet: The findings of this study suggest that repeated exposure of P. gingivalis in HCAEC induces the activation of proinflammatory and vasoconstrictor molecules that lead to endothelial dysfunction as a key mechanism of the onset and progression of arterial hypertension (HT) and atherosclerosis, which requires more research.

    Techniques: Cell Culture, Enzyme-linked Immunosorbent Assay

    Viability of HCAECs after repeated treatments with live- P. gingivalis ( A ) and P . gingivalis -LPS ( B ). The HCAECs were stimulated to repeated live- P. gingivalis (MOI 1:100 - 1:0,1) and P. gingivalis -LPS (1.0, 3.5 and 7.0 µg/mL) exposures, during 24 h. Cell viability was determined according to the fluorometric detection after reduction of resazurin in the resorufin product using AlamarBlue. 1% was considered our positive control of cell death. Percentage of cell viability with respect to the control. *Represents the statistical difference with respect to the control or without stimulus. (p

    Journal: Scientific Reports

    Article Title: Repeated Porphyromonas gingivalis W83 exposure leads to release pro-inflammatory cytokynes and angiotensin II in coronary artery endothelial cells

    doi: 10.1038/s41598-019-54259-y

    Figure Lengend Snippet: Viability of HCAECs after repeated treatments with live- P. gingivalis ( A ) and P . gingivalis -LPS ( B ). The HCAECs were stimulated to repeated live- P. gingivalis (MOI 1:100 - 1:0,1) and P. gingivalis -LPS (1.0, 3.5 and 7.0 µg/mL) exposures, during 24 h. Cell viability was determined according to the fluorometric detection after reduction of resazurin in the resorufin product using AlamarBlue. 1% was considered our positive control of cell death. Percentage of cell viability with respect to the control. *Represents the statistical difference with respect to the control or without stimulus. (p

    Article Snippet: The findings of this study suggest that repeated exposure of P. gingivalis in HCAEC induces the activation of proinflammatory and vasoconstrictor molecules that lead to endothelial dysfunction as a key mechanism of the onset and progression of arterial hypertension (HT) and atherosclerosis, which requires more research.

    Techniques: Positive Control

    mRNA expression levels in HCAEC stimulated with P. gingivalis -LPS or live- P. gingivalis . Monolayers of HCAEC cultured in 12-well plates were stimulated with P. gingivalis -LPS (1.0, 3.5, 7.0 µg/mL) or serial dilutions of P. gingivalis (MOI 1:100- 1:0,1) for 24 h under repeated exposure (+++) or single exposure (+). After stimulation, ( A ) AGTR1, ( B ) AGTR2, ( C ) IL-8, ( D ) IL-1β, ( E ) MCP-1, mRNA levels were measured by are expressed as the means by RT-qPCR. Results are expressed as the means ± SEM (n=3). Statiscal significance is represented as *p

    Journal: Scientific Reports

    Article Title: Repeated Porphyromonas gingivalis W83 exposure leads to release pro-inflammatory cytokynes and angiotensin II in coronary artery endothelial cells

    doi: 10.1038/s41598-019-54259-y

    Figure Lengend Snippet: mRNA expression levels in HCAEC stimulated with P. gingivalis -LPS or live- P. gingivalis . Monolayers of HCAEC cultured in 12-well plates were stimulated with P. gingivalis -LPS (1.0, 3.5, 7.0 µg/mL) or serial dilutions of P. gingivalis (MOI 1:100- 1:0,1) for 24 h under repeated exposure (+++) or single exposure (+). After stimulation, ( A ) AGTR1, ( B ) AGTR2, ( C ) IL-8, ( D ) IL-1β, ( E ) MCP-1, mRNA levels were measured by are expressed as the means by RT-qPCR. Results are expressed as the means ± SEM (n=3). Statiscal significance is represented as *p

    Article Snippet: The findings of this study suggest that repeated exposure of P. gingivalis in HCAEC induces the activation of proinflammatory and vasoconstrictor molecules that lead to endothelial dysfunction as a key mechanism of the onset and progression of arterial hypertension (HT) and atherosclerosis, which requires more research.

    Techniques: Expressing, Cell Culture, Quantitative RT-PCR

    Effects of OMV preexposure on TNF secretion in response to secondary stimulation with live P. gingivalis . (A) Human monocytes were prestimulated or not prestimulated with OMV for 24 h; after washing, cells were left unstimulated (medium) or restimulated

    Journal: Infection and Immunity

    Article Title: Porphyromonas gingivalis Outer Membrane Vesicles Induce Selective Tumor Necrosis Factor Tolerance in a Toll-Like Receptor 4- and mTOR-Dependent Manner

    doi: 10.1128/IAI.01390-15

    Figure Lengend Snippet: Effects of OMV preexposure on TNF secretion in response to secondary stimulation with live P. gingivalis . (A) Human monocytes were prestimulated or not prestimulated with OMV for 24 h; after washing, cells were left unstimulated (medium) or restimulated

    Article Snippet: We concluded that triggering of expression of surface TLR4 by P. gingivalis LPS in OMV is sufficient to prevent the release of TNF after secondary exposure of monocytes to P. gingivalis .

    Techniques:

    Role of fimbriae, gingipains, and endocytosis in TNF tolerance. (A) Monocytes were left unstimulated or prestimulated with Salmonella Minnesota LPS, P. gingivalis (ATCC 33277) OMV, or OMV of the nonfimbriated P. gingivalis W83 strain for 24 h. Cells were

    Journal: Infection and Immunity

    Article Title: Porphyromonas gingivalis Outer Membrane Vesicles Induce Selective Tumor Necrosis Factor Tolerance in a Toll-Like Receptor 4- and mTOR-Dependent Manner

    doi: 10.1128/IAI.01390-15

    Figure Lengend Snippet: Role of fimbriae, gingipains, and endocytosis in TNF tolerance. (A) Monocytes were left unstimulated or prestimulated with Salmonella Minnesota LPS, P. gingivalis (ATCC 33277) OMV, or OMV of the nonfimbriated P. gingivalis W83 strain for 24 h. Cells were

    Article Snippet: We concluded that triggering of expression of surface TLR4 by P. gingivalis LPS in OMV is sufficient to prevent the release of TNF after secondary exposure of monocytes to P. gingivalis .

    Techniques:

    Secretion of proinflammatory cytokines in response to P. gingivalis OMV. (A) Trans-electron microscopy of the purified OMV. The image shows a negative stain with 2% methylamine tungstate and is representative of the results of n = 6 experiments. (B and

    Journal: Infection and Immunity

    Article Title: Porphyromonas gingivalis Outer Membrane Vesicles Induce Selective Tumor Necrosis Factor Tolerance in a Toll-Like Receptor 4- and mTOR-Dependent Manner

    doi: 10.1128/IAI.01390-15

    Figure Lengend Snippet: Secretion of proinflammatory cytokines in response to P. gingivalis OMV. (A) Trans-electron microscopy of the purified OMV. The image shows a negative stain with 2% methylamine tungstate and is representative of the results of n = 6 experiments. (B and

    Article Snippet: We concluded that triggering of expression of surface TLR4 by P. gingivalis LPS in OMV is sufficient to prevent the release of TNF after secondary exposure of monocytes to P. gingivalis .

    Techniques: Electron Microscopy, Purification, Staining

    Role of TLR2 and TLR4 in OMV-mediated abrogation of the TNF response. (A to D) Human monocytes were prestimulated with OMV or not prestimulated; live P. gingivalis was used for secondary challenge after washing. The experiments were performed in the presence

    Journal: Infection and Immunity

    Article Title: Porphyromonas gingivalis Outer Membrane Vesicles Induce Selective Tumor Necrosis Factor Tolerance in a Toll-Like Receptor 4- and mTOR-Dependent Manner

    doi: 10.1128/IAI.01390-15

    Figure Lengend Snippet: Role of TLR2 and TLR4 in OMV-mediated abrogation of the TNF response. (A to D) Human monocytes were prestimulated with OMV or not prestimulated; live P. gingivalis was used for secondary challenge after washing. The experiments were performed in the presence

    Article Snippet: We concluded that triggering of expression of surface TLR4 by P. gingivalis LPS in OMV is sufficient to prevent the release of TNF after secondary exposure of monocytes to P. gingivalis .

    Techniques:

    P. gingivalis LPS-induced Wnt5a expression was inhibited by wedelolactone or dnIκBα. (A, B) THP-1 cells were pre-treated with the IKK inhibitor wedelolactone or the IKK inhibitor VII for 1 hr. The expression of Wnt5a mRNA was detected by real-time PCR. Relative expression levels of Wnt5a are presented. DMSO is a solvent control for both inhibitors. (C) THP-1 cells were transfected with a dominant-negative form of IκBα (dnIκBα) or the parent plasmid (pUSE) for 12 hrs. Fold induction of Wnt5a mRNA expression is shown. *p

    Journal: PLoS ONE

    Article Title: Modulation of Wnt5a Expression by Periodontopathic Bacteria

    doi: 10.1371/journal.pone.0034434

    Figure Lengend Snippet: P. gingivalis LPS-induced Wnt5a expression was inhibited by wedelolactone or dnIκBα. (A, B) THP-1 cells were pre-treated with the IKK inhibitor wedelolactone or the IKK inhibitor VII for 1 hr. The expression of Wnt5a mRNA was detected by real-time PCR. Relative expression levels of Wnt5a are presented. DMSO is a solvent control for both inhibitors. (C) THP-1 cells were transfected with a dominant-negative form of IκBα (dnIκBα) or the parent plasmid (pUSE) for 12 hrs. Fold induction of Wnt5a mRNA expression is shown. *p

    Article Snippet: In our study, primary cells including monocytes and gingival fibroblasts responded to P. gingivalis LPS in a similar manner to the cell lines (THP-1 and HGF-1).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Transfection, Dominant Negative Mutation, Plasmid Preparation

    Induction of Wnt5a expression is partly JAK/STAT dependent. (A–D) The expression of Wnt5a mRNA was detected by real-time PCR. Relative expression levels of Wnt5a are presented. (A, B) THP-1 cells were pre-treated with AG490 (A) or fludarabine (B) for 1 hr and then stimulated with P. gingivalis LPS for 4 hrs. DMSO is a solvent control for both inhibitors. (C) THP-1 cells were stimulated with P. gingivalis LPS for 4 hrs after transfection with STAT1 siRNA or control siRNA for 18 hrs. (D) THP-1 cells were pre-treated with STA21 for 1 hr and stimulated with P. gingivalis LPS for 4 hrs. *p

    Journal: PLoS ONE

    Article Title: Modulation of Wnt5a Expression by Periodontopathic Bacteria

    doi: 10.1371/journal.pone.0034434

    Figure Lengend Snippet: Induction of Wnt5a expression is partly JAK/STAT dependent. (A–D) The expression of Wnt5a mRNA was detected by real-time PCR. Relative expression levels of Wnt5a are presented. (A, B) THP-1 cells were pre-treated with AG490 (A) or fludarabine (B) for 1 hr and then stimulated with P. gingivalis LPS for 4 hrs. DMSO is a solvent control for both inhibitors. (C) THP-1 cells were stimulated with P. gingivalis LPS for 4 hrs after transfection with STAT1 siRNA or control siRNA for 18 hrs. (D) THP-1 cells were pre-treated with STA21 for 1 hr and stimulated with P. gingivalis LPS for 4 hrs. *p

    Article Snippet: In our study, primary cells including monocytes and gingival fibroblasts responded to P. gingivalis LPS in a similar manner to the cell lines (THP-1 and HGF-1).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Transfection

    Induction of Wnt5a expression is NF-κB dependent. (A) THP-1 cells were stimulated with E. coli LPS or P. gingivalis LPS for 30 mins or 4 hrs. Whole cell extracts were prepared and analyzed by Western blot by using antibodies against IκBα. β-actin served as the protein loading control. (B) THP-1 cells were stimulated with P. gingivalis LPS for 4 hrs after being transfected with NF-κB reporter plasmid for 12 hrs, and the transcription activity was assessed by luminometer. The activity is represented by the relative luciferase activity. (C) Nuclear extracts were prepared and EMSA was performed with γ 32 P-labeled oligonucleotides representing the NF-κB consensus sequence as a probe. Anti-p65 antibody was used for supershift assays. The lower arrow shows the DNA-protein complex, and the upper arrow shows the supershifted band. *p

    Journal: PLoS ONE

    Article Title: Modulation of Wnt5a Expression by Periodontopathic Bacteria

    doi: 10.1371/journal.pone.0034434

    Figure Lengend Snippet: Induction of Wnt5a expression is NF-κB dependent. (A) THP-1 cells were stimulated with E. coli LPS or P. gingivalis LPS for 30 mins or 4 hrs. Whole cell extracts were prepared and analyzed by Western blot by using antibodies against IκBα. β-actin served as the protein loading control. (B) THP-1 cells were stimulated with P. gingivalis LPS for 4 hrs after being transfected with NF-κB reporter plasmid for 12 hrs, and the transcription activity was assessed by luminometer. The activity is represented by the relative luciferase activity. (C) Nuclear extracts were prepared and EMSA was performed with γ 32 P-labeled oligonucleotides representing the NF-κB consensus sequence as a probe. Anti-p65 antibody was used for supershift assays. The lower arrow shows the DNA-protein complex, and the upper arrow shows the supershifted band. *p

    Article Snippet: In our study, primary cells including monocytes and gingival fibroblasts responded to P. gingivalis LPS in a similar manner to the cell lines (THP-1 and HGF-1).

    Techniques: Expressing, Western Blot, Transfection, Plasmid Preparation, Activity Assay, Luciferase, Labeling, Sequencing

    The expression of Wnt5a was increased by co-stimulation with P. gingivalis LPS and IFN-γ. (A–C) The expression of Wnt5a mRNA was detected by real-time PCR. Relative expression levels of Wnt5a are presented. (A) THP-1 cells were stimulated with 10 ng/ml of IL-6, 10 ng/ml of IFN-β or 10 ng/ml of IFN-γ with or without P. gingivalis LPS for 4 hrs. (B) THP-1 cells were transfected with wild-type STAT1 expression vector or parent plasmid 12 hrs prior to stimulation with P. gingivalis LPS/IFN-γ for 4 hrs. (C) THP-1 cells were stimulated with P. gingivalis LPS for 4 hrs after being transfected with STAT1 siRNA or control siRNA for 18 hrs. *p

    Journal: PLoS ONE

    Article Title: Modulation of Wnt5a Expression by Periodontopathic Bacteria

    doi: 10.1371/journal.pone.0034434

    Figure Lengend Snippet: The expression of Wnt5a was increased by co-stimulation with P. gingivalis LPS and IFN-γ. (A–C) The expression of Wnt5a mRNA was detected by real-time PCR. Relative expression levels of Wnt5a are presented. (A) THP-1 cells were stimulated with 10 ng/ml of IL-6, 10 ng/ml of IFN-β or 10 ng/ml of IFN-γ with or without P. gingivalis LPS for 4 hrs. (B) THP-1 cells were transfected with wild-type STAT1 expression vector or parent plasmid 12 hrs prior to stimulation with P. gingivalis LPS/IFN-γ for 4 hrs. (C) THP-1 cells were stimulated with P. gingivalis LPS for 4 hrs after being transfected with STAT1 siRNA or control siRNA for 18 hrs. *p

    Article Snippet: In our study, primary cells including monocytes and gingival fibroblasts responded to P. gingivalis LPS in a similar manner to the cell lines (THP-1 and HGF-1).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Transfection, Plasmid Preparation

    Wnt5a was specifically up-regulated in THP-1 cells by P. gingivalis LPS. (A) HGF-1 and THP-1 cells were stimulated with A. actinomycetemcomitans sonicated extract, P. gingivalis sonicated extract, P. gingivalis LPS, and TNF-α for 4 hrs, and the expression of Wnt5a mRNA was detected by RT-PCR. PCR products were electrophoresed on a 1.5% agarose gel and visualized by UV illumination. GAPDH served as the internal control. (B) THP-1 cells were stimulated with 1 µg/ml of P. gingivalis LPS for 0.5, 2, 4, 12, or 24 hrs, and the expression of Wnt5a mRNA was detected by real-time PCR. Relative expression levels of Wnt5a are shown. (C) THP-1 cells were stimulated with 0.01–10 µg/ml of P. gingivalis LPS (black bars) or E. coli LPS (gray bars) for 4 hrs, and then the expression of Wnt5a mRNA was detected by real-time PCR. Relative expression levels of Wnt5a are presented. (D) THP-1 cells were stimulated with E. coli 055:B5 LPS (middle and right upper panels) or P. gingivalis LPS (middle and right lower panels) for 30 min or 4 hrs, and then the expression of surface TLR2 and TLR4 protein was determined by flow cytometry. Left upper panel shows no-staining condition, and left lower panel shows un-stimulated condition with staining. (E, F, G, H) The expression of Wnt5a mRNA was detected by real-time PCR. Relative expression levels of Wnt5a mRNA are shown. (E) THP-1 cells were stimulated with 10 4 –10 7 cells/ml of live P. gingivalis for 4 hrs. (F, G) Primary human gingival fibroblasts (HGF) and human monocytes were stimulated with 1 µg/ml of P. gingivalis LPS for 4 hrs. Monocytes were pretreated with the NF-κB inhibitor MG132 for 1 hr. Here we describe typical dates of three samples. (H) THP-1 cells were stimulated with P. gingivalis LPS for 4 hrs after being transfected with TLR2 siRNA, TLR4 siRNA or control siRNA for 72 hrs. *p

    Journal: PLoS ONE

    Article Title: Modulation of Wnt5a Expression by Periodontopathic Bacteria

    doi: 10.1371/journal.pone.0034434

    Figure Lengend Snippet: Wnt5a was specifically up-regulated in THP-1 cells by P. gingivalis LPS. (A) HGF-1 and THP-1 cells were stimulated with A. actinomycetemcomitans sonicated extract, P. gingivalis sonicated extract, P. gingivalis LPS, and TNF-α for 4 hrs, and the expression of Wnt5a mRNA was detected by RT-PCR. PCR products were electrophoresed on a 1.5% agarose gel and visualized by UV illumination. GAPDH served as the internal control. (B) THP-1 cells were stimulated with 1 µg/ml of P. gingivalis LPS for 0.5, 2, 4, 12, or 24 hrs, and the expression of Wnt5a mRNA was detected by real-time PCR. Relative expression levels of Wnt5a are shown. (C) THP-1 cells were stimulated with 0.01–10 µg/ml of P. gingivalis LPS (black bars) or E. coli LPS (gray bars) for 4 hrs, and then the expression of Wnt5a mRNA was detected by real-time PCR. Relative expression levels of Wnt5a are presented. (D) THP-1 cells were stimulated with E. coli 055:B5 LPS (middle and right upper panels) or P. gingivalis LPS (middle and right lower panels) for 30 min or 4 hrs, and then the expression of surface TLR2 and TLR4 protein was determined by flow cytometry. Left upper panel shows no-staining condition, and left lower panel shows un-stimulated condition with staining. (E, F, G, H) The expression of Wnt5a mRNA was detected by real-time PCR. Relative expression levels of Wnt5a mRNA are shown. (E) THP-1 cells were stimulated with 10 4 –10 7 cells/ml of live P. gingivalis for 4 hrs. (F, G) Primary human gingival fibroblasts (HGF) and human monocytes were stimulated with 1 µg/ml of P. gingivalis LPS for 4 hrs. Monocytes were pretreated with the NF-κB inhibitor MG132 for 1 hr. Here we describe typical dates of three samples. (H) THP-1 cells were stimulated with P. gingivalis LPS for 4 hrs after being transfected with TLR2 siRNA, TLR4 siRNA or control siRNA for 72 hrs. *p

    Article Snippet: In our study, primary cells including monocytes and gingival fibroblasts responded to P. gingivalis LPS in a similar manner to the cell lines (THP-1 and HGF-1).

    Techniques: Sonication, Expressing, Reverse Transcription Polymerase Chain Reaction, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Real-time Polymerase Chain Reaction, Flow Cytometry, Cytometry, Staining, Transfection

    Comparison of proinflammatory cytokine production in WT and TLR9 −/− macrophages (A, B) and splenocytes (C, D) in response to different TLR agonists. The cells were stimulated with ODN 1668 (TLR9 agonist; 100 ng/μl), P. gingivalis LPS (TLR4 agonist; 10 ng/μl), E. coli LPS (TLR4 agonist; 10 ng/μl), or Pam3Cys (TLR2 agonist; 1 ng/μl) for 24 h. Cell-free supernatants were analyzed for the presence of IL-6 and TNF using ELISA. The results shown are representative of at least 3 independent experiments that were run in triplicates. The data shown are the mean results ± SD ( n = 9). Comparisons between WT and KO cells were performed using the unpaired Student t test. *, P

    Journal: Infection and Immunity

    Article Title: Toll-Like Receptor 9-Mediated Inflammation Triggers Alveolar Bone Loss in Experimental Murine Periodontitis

    doi: 10.1128/IAI.00424-15

    Figure Lengend Snippet: Comparison of proinflammatory cytokine production in WT and TLR9 −/− macrophages (A, B) and splenocytes (C, D) in response to different TLR agonists. The cells were stimulated with ODN 1668 (TLR9 agonist; 100 ng/μl), P. gingivalis LPS (TLR4 agonist; 10 ng/μl), E. coli LPS (TLR4 agonist; 10 ng/μl), or Pam3Cys (TLR2 agonist; 1 ng/μl) for 24 h. Cell-free supernatants were analyzed for the presence of IL-6 and TNF using ELISA. The results shown are representative of at least 3 independent experiments that were run in triplicates. The data shown are the mean results ± SD ( n = 9). Comparisons between WT and KO cells were performed using the unpaired Student t test. *, P

    Article Snippet: The cells were stimulated with ODN 1668 (TLR9 agonist, 100 ng/μl; InviVogen), P. gingivalis (multiplicity of infection [MOI] of 1:100), P. gingivalis DNA (100 ng/μl), P. gingivalis lipopolysaccharide (LPS) (10 ng/μl; InviVogen), Escherichia coli LPS (10 ng/μl; InviVogen), and Pam3Cys (1 ng/μl; InviVogen) for 24 h. Inflammatory cytokine levels (IL-6 and TNF) were determined in cell-free culture supernatants using ELISAs (eBiosciences).

    Techniques: Enzyme-linked Immunosorbent Assay

    Cytokine expression levels upon stimulation of U937 macrophages with E. coli (0.1 μg/ml), P. gingivalis , and T. forsythia (1 μg/ml) LPS in the presence or absence of FCS. In both qPCR (A) and ELISA (B), P. gingivalis LPS did not show serum dependence for cytokine production. E. coli and T. forsythia LPS caused a substantially higher release of IL-1β, IL-6, and TNF-α in the presence of serum. The data are mean values ± SDs of three wells originating from one representative experiment. A similar tendency was observed in other experiments. *, significantly different from the control at P

    Journal: Clinical and Vaccine Immunology : CVI

    Article Title: Structure and Immunogenicity of the Rough-Type Lipopolysaccharide from the Periodontal Pathogen Tannerella forsythia

    doi: 10.1128/CVI.00139-13

    Figure Lengend Snippet: Cytokine expression levels upon stimulation of U937 macrophages with E. coli (0.1 μg/ml), P. gingivalis , and T. forsythia (1 μg/ml) LPS in the presence or absence of FCS. In both qPCR (A) and ELISA (B), P. gingivalis LPS did not show serum dependence for cytokine production. E. coli and T. forsythia LPS caused a substantially higher release of IL-1β, IL-6, and TNF-α in the presence of serum. The data are mean values ± SDs of three wells originating from one representative experiment. A similar tendency was observed in other experiments. *, significantly different from the control at P

    Article Snippet: Cells were stimulated with T. forsythia LPS at final concentrations ranging from 0.01 to 10 μg/ml for 24 h. Cells stimulated with P. gingivalis LPS (1 μg/ml; Life Technologies) and Escherichia coli O111:B4 LPS (100 ng/ml; Life Technologies) were taken as positive controls, and nonstimulated cells were taken as a negative control.

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

    LPS from P. gingivalis ( P.g. ) stimulates bone resorption, osteoclast formation, and expression of osteoclastic and osteoclastogenic genes in organ cultures of neonatal mouse parietal bones. A–C , LPS P. gingivalis time- and concentration-dependently

    Journal: The Journal of Biological Chemistry

    Article Title: Porphyromonas gingivalis Stimulates Bone Resorption by Enhancing RANKL (Receptor Activator of NF-κB Ligand) through Activation of Toll-like Receptor 2 in Osteoblasts *

    doi: 10.1074/jbc.M115.655787

    Figure Lengend Snippet: LPS from P. gingivalis ( P.g. ) stimulates bone resorption, osteoclast formation, and expression of osteoclastic and osteoclastogenic genes in organ cultures of neonatal mouse parietal bones. A–C , LPS P. gingivalis time- and concentration-dependently

    Article Snippet: Recombinant mouse cytokines and neutralizing antibodies and Quantikine® ELISA kits for RANKL and OPG were from R & D Systems; BMS-345541 and Celastrol were from Sigma; α-minimum essential medium, fetal calf serum, zoledronic acid, and indomethacin were from Invitrogen; 45 CaCl2 was from Amersham Biosciences; oligonucleotide primers and probes were from Invitrogen or Applied Biosystems; LPS P. gingivalis (version 10G20-MT) and other TLR2 and TLR4 agonists and primers were from InvivoGen and R & D Systems; RatLapsTM CTX ELISA kit was from Immunodiagnostic Systems; prostaglandin E2 125 I-RIA® kit was from PerkinElmer Life Sciences; RNAqueous-4 PCR® kit was from Ambion; High Capacity cDNA Reverse Transcription kit was from Applied Biosystems; Kapa2GTM Robust HotStart PCR kit and KapaTM Probe Fast qPCR kit were from Kapa Biosystems; TaqMan® Fast Advanced Master Mix was from Life Technologies; RNAlater®, RNeasy®, and Cignal Lenti Reporter Assay® kits were from Qiagen; Luciferase Assay System was from Promega.

    Techniques: Expressing, Concentration Assay

    Five different TLR2 agonists enhanced Tnfsf11 mRNA expression in mouse parietal osteoblasts by a mechanism dependent on TLR2, MyD88, and NF-κB but independent on cytokine formation. A , LPS P. gingivalis ( P.g. ) and Pam2 up-regulated Tlr2 without

    Journal: The Journal of Biological Chemistry

    Article Title: Porphyromonas gingivalis Stimulates Bone Resorption by Enhancing RANKL (Receptor Activator of NF-κB Ligand) through Activation of Toll-like Receptor 2 in Osteoblasts *

    doi: 10.1074/jbc.M115.655787

    Figure Lengend Snippet: Five different TLR2 agonists enhanced Tnfsf11 mRNA expression in mouse parietal osteoblasts by a mechanism dependent on TLR2, MyD88, and NF-κB but independent on cytokine formation. A , LPS P. gingivalis ( P.g. ) and Pam2 up-regulated Tlr2 without

    Article Snippet: Recombinant mouse cytokines and neutralizing antibodies and Quantikine® ELISA kits for RANKL and OPG were from R & D Systems; BMS-345541 and Celastrol were from Sigma; α-minimum essential medium, fetal calf serum, zoledronic acid, and indomethacin were from Invitrogen; 45 CaCl2 was from Amersham Biosciences; oligonucleotide primers and probes were from Invitrogen or Applied Biosystems; LPS P. gingivalis (version 10G20-MT) and other TLR2 and TLR4 agonists and primers were from InvivoGen and R & D Systems; RatLapsTM CTX ELISA kit was from Immunodiagnostic Systems; prostaglandin E2 125 I-RIA® kit was from PerkinElmer Life Sciences; RNAqueous-4 PCR® kit was from Ambion; High Capacity cDNA Reverse Transcription kit was from Applied Biosystems; Kapa2GTM Robust HotStart PCR kit and KapaTM Probe Fast qPCR kit were from Kapa Biosystems; TaqMan® Fast Advanced Master Mix was from Life Technologies; RNAlater®, RNeasy®, and Cignal Lenti Reporter Assay® kits were from Qiagen; Luciferase Assay System was from Promega.

    Techniques: Expressing

    Injection of LPS from P. gingivalis ( P.g. ) and the TLR2 agonist Pam2 above skull bones stimulates osteoclast formation, expression of osteoclastic and osteoclastogenic genes, and bone loss in skull bones from 5-week-old mice. A and B , LPS P. gingivalis

    Journal: The Journal of Biological Chemistry

    Article Title: Porphyromonas gingivalis Stimulates Bone Resorption by Enhancing RANKL (Receptor Activator of NF-κB Ligand) through Activation of Toll-like Receptor 2 in Osteoblasts *

    doi: 10.1074/jbc.M115.655787

    Figure Lengend Snippet: Injection of LPS from P. gingivalis ( P.g. ) and the TLR2 agonist Pam2 above skull bones stimulates osteoclast formation, expression of osteoclastic and osteoclastogenic genes, and bone loss in skull bones from 5-week-old mice. A and B , LPS P. gingivalis

    Article Snippet: Recombinant mouse cytokines and neutralizing antibodies and Quantikine® ELISA kits for RANKL and OPG were from R & D Systems; BMS-345541 and Celastrol were from Sigma; α-minimum essential medium, fetal calf serum, zoledronic acid, and indomethacin were from Invitrogen; 45 CaCl2 was from Amersham Biosciences; oligonucleotide primers and probes were from Invitrogen or Applied Biosystems; LPS P. gingivalis (version 10G20-MT) and other TLR2 and TLR4 agonists and primers were from InvivoGen and R & D Systems; RatLapsTM CTX ELISA kit was from Immunodiagnostic Systems; prostaglandin E2 125 I-RIA® kit was from PerkinElmer Life Sciences; RNAqueous-4 PCR® kit was from Ambion; High Capacity cDNA Reverse Transcription kit was from Applied Biosystems; Kapa2GTM Robust HotStart PCR kit and KapaTM Probe Fast qPCR kit were from Kapa Biosystems; TaqMan® Fast Advanced Master Mix was from Life Technologies; RNAlater®, RNeasy®, and Cignal Lenti Reporter Assay® kits were from Qiagen; Luciferase Assay System was from Promega.

    Techniques: Injection, Expressing, Mouse Assay

    The stimulatory effect on bone resorption in neonatal mouse parietal bones by LPS from P. gingivalis ( P.g. ) is dependent on increased RANKL. A , LPS P. gingivalis enhanced the mRNA expression of Tnfrsf11a , Tnfsf11 , Csf1 , Csf1r , and Oscar without affecting

    Journal: The Journal of Biological Chemistry

    Article Title: Porphyromonas gingivalis Stimulates Bone Resorption by Enhancing RANKL (Receptor Activator of NF-κB Ligand) through Activation of Toll-like Receptor 2 in Osteoblasts *

    doi: 10.1074/jbc.M115.655787

    Figure Lengend Snippet: The stimulatory effect on bone resorption in neonatal mouse parietal bones by LPS from P. gingivalis ( P.g. ) is dependent on increased RANKL. A , LPS P. gingivalis enhanced the mRNA expression of Tnfrsf11a , Tnfsf11 , Csf1 , Csf1r , and Oscar without affecting

    Article Snippet: Recombinant mouse cytokines and neutralizing antibodies and Quantikine® ELISA kits for RANKL and OPG were from R & D Systems; BMS-345541 and Celastrol were from Sigma; α-minimum essential medium, fetal calf serum, zoledronic acid, and indomethacin were from Invitrogen; 45 CaCl2 was from Amersham Biosciences; oligonucleotide primers and probes were from Invitrogen or Applied Biosystems; LPS P. gingivalis (version 10G20-MT) and other TLR2 and TLR4 agonists and primers were from InvivoGen and R & D Systems; RatLapsTM CTX ELISA kit was from Immunodiagnostic Systems; prostaglandin E2 125 I-RIA® kit was from PerkinElmer Life Sciences; RNAqueous-4 PCR® kit was from Ambion; High Capacity cDNA Reverse Transcription kit was from Applied Biosystems; Kapa2GTM Robust HotStart PCR kit and KapaTM Probe Fast qPCR kit were from Kapa Biosystems; TaqMan® Fast Advanced Master Mix was from Life Technologies; RNAlater®, RNeasy®, and Cignal Lenti Reporter Assay® kits were from Qiagen; Luciferase Assay System was from Promega.

    Techniques: Expressing

    Anti-microbial activity of clinical working concentrations of PHMG-P (1%) and CHX (0.2%) against perio- and cariogenic bacteria ( P. gingivalis , A. actinomycetemcomitans , S. mutans and L. acidophilus ).

    Journal: Annals of Clinical Microbiology and Antimicrobials

    Article Title: Antimicrobial activity of polyhexamethylene guanidine phosphate in comparison to chlorhexidine using the quantitative suspension method

    doi: 10.1186/s12941-015-0097-x

    Figure Lengend Snippet: Anti-microbial activity of clinical working concentrations of PHMG-P (1%) and CHX (0.2%) against perio- and cariogenic bacteria ( P. gingivalis , A. actinomycetemcomitans , S. mutans and L. acidophilus ).

    Article Snippet: The test-suspension of P. gingivalis with OD 0.73–0.75 was prepared in peptone yeast glucose broth (Becton–Dickinson).

    Techniques: Activity Assay

    Anti-microbial activity of highly diluted PHMG-P (0.05%) and CHX (0.05%) against perio- and cariogenic bacteria ( P. gingivalis , A. actinomycetemcomitans , S. mutans and L. acidophilus ).

    Journal: Annals of Clinical Microbiology and Antimicrobials

    Article Title: Antimicrobial activity of polyhexamethylene guanidine phosphate in comparison to chlorhexidine using the quantitative suspension method

    doi: 10.1186/s12941-015-0097-x

    Figure Lengend Snippet: Anti-microbial activity of highly diluted PHMG-P (0.05%) and CHX (0.05%) against perio- and cariogenic bacteria ( P. gingivalis , A. actinomycetemcomitans , S. mutans and L. acidophilus ).

    Article Snippet: The test-suspension of P. gingivalis with OD 0.73–0.75 was prepared in peptone yeast glucose broth (Becton–Dickinson).

    Techniques: Activity Assay

    Persistence of P. gingivalis within HCAE cells over 8 h. HCAE cells were preincubated for 1 h at 37°C with either fresh antibiotic-free EGM-2 (solid bars), 10 mM 3-methyladenine (open bars), or 10 nM wortmannin (checkered bars) ( n = 3). P. gingivalis 381 cells were incubated with EGM-2 with or without the appropriate inhibitor. The number of CFU of HCAE cells infected with P. gingivalis in the absence of autophagy inhibitors grew over the 8 h, whereas the number of CFU of HCAE cells infected with P. gingivalis in the presence of either autophagy inhibitor declined over the 8 h. The number of CFU for E. coli MC1061, the negative control, at 2.5 h postinfection was

    Journal: Infection and Immunity

    Article Title: Porphyromonas gingivalis Traffics to Autophagosomes in Human Coronary Artery Endothelial Cells

    doi: 10.1128/IAI.69.9.5698-5708.2001

    Figure Lengend Snippet: Persistence of P. gingivalis within HCAE cells over 8 h. HCAE cells were preincubated for 1 h at 37°C with either fresh antibiotic-free EGM-2 (solid bars), 10 mM 3-methyladenine (open bars), or 10 nM wortmannin (checkered bars) ( n = 3). P. gingivalis 381 cells were incubated with EGM-2 with or without the appropriate inhibitor. The number of CFU of HCAE cells infected with P. gingivalis in the absence of autophagy inhibitors grew over the 8 h, whereas the number of CFU of HCAE cells infected with P. gingivalis in the presence of either autophagy inhibitor declined over the 8 h. The number of CFU for E. coli MC1061, the negative control, at 2.5 h postinfection was

    Article Snippet: Dilutions of the lysates of cells infected with P. gingivalis were plated in triplicate on tryptic soy agar (Difco) plates supplemented with 5.0% sheep blood, 0.5% yeast extract, hemin (5 μg/ml), and vitamin K (5 μg/ml) and were cultured anaerobically.

    Techniques: Incubation, Infection, Negative Control

    Internalized P. gingivalis in HCAE cells. HCAE cells were infected with P. gingivalis in the presence (A, C, and D) and absence (B) of wortmannin. After 90 min of infection, P. gingivalis could be observed in vacuoles within the cytoplasm of HCAE cells (A). These vacuoles were bound by one or two membranes (arrows) and contained undegraded vesicles and cytoplasmic ground substance (C and D). After 30 min of infection of wortmannin-treated HCAE cells, P. gingivalis appears to be in the process of degradation and is within vacuoles that resemble lysosomes (B).

    Journal: Infection and Immunity

    Article Title: Porphyromonas gingivalis Traffics to Autophagosomes in Human Coronary Artery Endothelial Cells

    doi: 10.1128/IAI.69.9.5698-5708.2001

    Figure Lengend Snippet: Internalized P. gingivalis in HCAE cells. HCAE cells were infected with P. gingivalis in the presence (A, C, and D) and absence (B) of wortmannin. After 90 min of infection, P. gingivalis could be observed in vacuoles within the cytoplasm of HCAE cells (A). These vacuoles were bound by one or two membranes (arrows) and contained undegraded vesicles and cytoplasmic ground substance (C and D). After 30 min of infection of wortmannin-treated HCAE cells, P. gingivalis appears to be in the process of degradation and is within vacuoles that resemble lysosomes (B).

    Article Snippet: Dilutions of the lysates of cells infected with P. gingivalis were plated in triplicate on tryptic soy agar (Difco) plates supplemented with 5.0% sheep blood, 0.5% yeast extract, hemin (5 μg/ml), and vitamin K (5 μg/ml) and were cultured anaerobically.

    Techniques: Infection

    Localization of HsGsa7p and cathepsin L to vacuoles containing P. gingivalis in HCAE cells using deconvolution microscopy. (A) P. gingivalis colocalized with HsGsa7p (arrow). (B) In the presence of wortmannin, P. gingivalis did not colocalize with HsGsa7p (arrowhead). HsGsa7p resided in a Golgi-like vacuole in the wortmannin-treated HCAE cells (arrowhead). (C) P. gingivalis did not colocalize with cathepsin L (arrowhead). (D) P. gingivalis colocalized with cathepsin L (arrow) in the presence of wortmannin. Abbreviations: Gsa7p, HsGsa7p; W, wortmannin; CathL, cathepsin L; Pg, P. gingivalis ; N, nucleus. Bar, 15 μm.

    Journal: Infection and Immunity

    Article Title: Porphyromonas gingivalis Traffics to Autophagosomes in Human Coronary Artery Endothelial Cells

    doi: 10.1128/IAI.69.9.5698-5708.2001

    Figure Lengend Snippet: Localization of HsGsa7p and cathepsin L to vacuoles containing P. gingivalis in HCAE cells using deconvolution microscopy. (A) P. gingivalis colocalized with HsGsa7p (arrow). (B) In the presence of wortmannin, P. gingivalis did not colocalize with HsGsa7p (arrowhead). HsGsa7p resided in a Golgi-like vacuole in the wortmannin-treated HCAE cells (arrowhead). (C) P. gingivalis did not colocalize with cathepsin L (arrowhead). (D) P. gingivalis colocalized with cathepsin L (arrow) in the presence of wortmannin. Abbreviations: Gsa7p, HsGsa7p; W, wortmannin; CathL, cathepsin L; Pg, P. gingivalis ; N, nucleus. Bar, 15 μm.

    Article Snippet: Dilutions of the lysates of cells infected with P. gingivalis were plated in triplicate on tryptic soy agar (Difco) plates supplemented with 5.0% sheep blood, 0.5% yeast extract, hemin (5 μg/ml), and vitamin K (5 μg/ml) and were cultured anaerobically.

    Techniques: Microscopy

    Localization of BiP, LGP120, Rab5, and MPR to vacuoles containing P. gingivalis in HCAE cells. At 90 min postinfection, the P. gingivalis (Pg) vacuoles contained BiP (A) and LGP120 (B) (arrows). (C) At 35 min after infection, P. gingivalis colocalized with Rab5 (arrow). (D) MPR was absent from a majority of the vacuoles observed up to 2 h postinfection (arrowheads indicate colocalization). Bar, 15 μm.

    Journal: Infection and Immunity

    Article Title: Porphyromonas gingivalis Traffics to Autophagosomes in Human Coronary Artery Endothelial Cells

    doi: 10.1128/IAI.69.9.5698-5708.2001

    Figure Lengend Snippet: Localization of BiP, LGP120, Rab5, and MPR to vacuoles containing P. gingivalis in HCAE cells. At 90 min postinfection, the P. gingivalis (Pg) vacuoles contained BiP (A) and LGP120 (B) (arrows). (C) At 35 min after infection, P. gingivalis colocalized with Rab5 (arrow). (D) MPR was absent from a majority of the vacuoles observed up to 2 h postinfection (arrowheads indicate colocalization). Bar, 15 μm.

    Article Snippet: Dilutions of the lysates of cells infected with P. gingivalis were plated in triplicate on tryptic soy agar (Difco) plates supplemented with 5.0% sheep blood, 0.5% yeast extract, hemin (5 μg/ml), and vitamin K (5 μg/ml) and were cultured anaerobically.

    Techniques: Infection

    Colocalization of P. gingivalis with protein markers of the autophagic pathway. HCAE cells were incubated with P. gingivalis 381 in the absence (solid bars) and presence (open bars) of 10 nM wortmannin for 15 to 120 min. The data represent the percentage of P. gingivalis vacuoles that contained BiP (A), HsGsa7p (B), LGP120 (C), and cathepsin L (D), expressed as the mean + the standard deviation (error bars). The data suggest that the P. gingivalis vacuole first acquires HsGsa7p and then BiP and LGP120; however, the vacuole fails to acquire cathepsin L. In the presence of wortmannin, the vacuole does not acquire HsGsa7p or BiP but rather acquires cathepsin L rapidly. The effects of wortmannin at each time point were evaluated by Student's t test utilizing Bonferroni's correction. Statistical significance (∗) was achieved when P was

    Journal: Infection and Immunity

    Article Title: Porphyromonas gingivalis Traffics to Autophagosomes in Human Coronary Artery Endothelial Cells

    doi: 10.1128/IAI.69.9.5698-5708.2001

    Figure Lengend Snippet: Colocalization of P. gingivalis with protein markers of the autophagic pathway. HCAE cells were incubated with P. gingivalis 381 in the absence (solid bars) and presence (open bars) of 10 nM wortmannin for 15 to 120 min. The data represent the percentage of P. gingivalis vacuoles that contained BiP (A), HsGsa7p (B), LGP120 (C), and cathepsin L (D), expressed as the mean + the standard deviation (error bars). The data suggest that the P. gingivalis vacuole first acquires HsGsa7p and then BiP and LGP120; however, the vacuole fails to acquire cathepsin L. In the presence of wortmannin, the vacuole does not acquire HsGsa7p or BiP but rather acquires cathepsin L rapidly. The effects of wortmannin at each time point were evaluated by Student's t test utilizing Bonferroni's correction. Statistical significance (∗) was achieved when P was

    Article Snippet: Dilutions of the lysates of cells infected with P. gingivalis were plated in triplicate on tryptic soy agar (Difco) plates supplemented with 5.0% sheep blood, 0.5% yeast extract, hemin (5 μg/ml), and vitamin K (5 μg/ml) and were cultured anaerobically.

    Techniques: Incubation, Standard Deviation

    Model of P. gingivalis trafficking in HCAE cells. P. gingivalis is initially found within an early endosome after internalization. (A) The bacteria promote their own entry into the autophagic pathway. The P. gingivalis 381-containing vacuole acquires BiP and later LGP120. However, this vacuole does not acquire cathepsin L. (B) Upon inhibition of autophagy with wortmannin, P. gingivalis enters the endocytic pathway. The vacuole matures into a late endosome and then a lysosome, as characterized by the presence of Rap1 and cathepsin L.

    Journal: Infection and Immunity

    Article Title: Porphyromonas gingivalis Traffics to Autophagosomes in Human Coronary Artery Endothelial Cells

    doi: 10.1128/IAI.69.9.5698-5708.2001

    Figure Lengend Snippet: Model of P. gingivalis trafficking in HCAE cells. P. gingivalis is initially found within an early endosome after internalization. (A) The bacteria promote their own entry into the autophagic pathway. The P. gingivalis 381-containing vacuole acquires BiP and later LGP120. However, this vacuole does not acquire cathepsin L. (B) Upon inhibition of autophagy with wortmannin, P. gingivalis enters the endocytic pathway. The vacuole matures into a late endosome and then a lysosome, as characterized by the presence of Rap1 and cathepsin L.

    Article Snippet: Dilutions of the lysates of cells infected with P. gingivalis were plated in triplicate on tryptic soy agar (Difco) plates supplemented with 5.0% sheep blood, 0.5% yeast extract, hemin (5 μg/ml), and vitamin K (5 μg/ml) and were cultured anaerobically.

    Techniques: Inhibition

    Colocalization of P. gingivalis with protein markers of the endocytic pathway. HCAE cells were incubated with P. gingivalis 381 in the absence (solid bars) and presence (open bars) of 10 nM wortmannin for 15 to 120 min. The data represent the percentage of P. gingivalis vacuoles that contained Rab5 (A) and MPR (B), expressed as the mean + the standard deviation (error bars). The data suggest that the P. gingivalis vacuole acquires Rab5 early but does not acquire the late endocytic marker MPR. In the presence of wortmannin, a higher percentage of P. gingivalis vacuoles acquire MPR. The effects of wortmannin at each time point were evaluated by Student's t test utilizing Bonferroni's correction. Statistical significance (*) was achieved when P was

    Journal: Infection and Immunity

    Article Title: Porphyromonas gingivalis Traffics to Autophagosomes in Human Coronary Artery Endothelial Cells

    doi: 10.1128/IAI.69.9.5698-5708.2001

    Figure Lengend Snippet: Colocalization of P. gingivalis with protein markers of the endocytic pathway. HCAE cells were incubated with P. gingivalis 381 in the absence (solid bars) and presence (open bars) of 10 nM wortmannin for 15 to 120 min. The data represent the percentage of P. gingivalis vacuoles that contained Rab5 (A) and MPR (B), expressed as the mean + the standard deviation (error bars). The data suggest that the P. gingivalis vacuole acquires Rab5 early but does not acquire the late endocytic marker MPR. In the presence of wortmannin, a higher percentage of P. gingivalis vacuoles acquire MPR. The effects of wortmannin at each time point were evaluated by Student's t test utilizing Bonferroni's correction. Statistical significance (*) was achieved when P was

    Article Snippet: Dilutions of the lysates of cells infected with P. gingivalis were plated in triplicate on tryptic soy agar (Difco) plates supplemented with 5.0% sheep blood, 0.5% yeast extract, hemin (5 μg/ml), and vitamin K (5 μg/ml) and were cultured anaerobically.

    Techniques: Incubation, Standard Deviation, Marker

    Time-kill curves for P. gingivalis JCM12257 (a) and A. actinomycetemcomitans JCM8577 (b) following exposure to 0.9% NaCl (control), 0.2% CHX, and NBW3. The number of CFUs/mL of P. gingivalis exposed to 0.2% CHX did not drop to below the lower limit of detection (

    Journal: Science and Technology of Advanced Materials

    Article Title: Effects of ozone nano-bubble water on periodontopathic bacteria and oral cells - in vitro studies

    doi: 10.1088/1468-6996/15/5/055003

    Figure Lengend Snippet: Time-kill curves for P. gingivalis JCM12257 (a) and A. actinomycetemcomitans JCM8577 (b) following exposure to 0.9% NaCl (control), 0.2% CHX, and NBW3. The number of CFUs/mL of P. gingivalis exposed to 0.2% CHX did not drop to below the lower limit of detection (

    Article Snippet: P. gingivalis was grown anaerobically on modified GAM agar (Nissui Pharmaceutical Co., Ltd, Ueno, Tokyo, Japan) for 3 days at 37 °C.

    Techniques:

    fim region able to swap during natural competence despite limited conservation regions. (A.) Complete fim gene region ( fimA-fimE ) and flanking genes are shown for sequenced P. gingivalis strains 33277 fimA (I ), TDC60 fimA (II ), and W83 fimA (IV ). Overall sequence conservation between all strains is displayed. Green indicates the most conservation while yellow, orange, and finally red signifies the least DNA conservation between strains. Distances are shown in base pairs. Figure modified from CLC Genomic Workbench 6, accession numbers 33277(NC_010729), TD60 (NC_015571), and W83 (NC_002950). (B.) Chromosomal preparations from a random sample of ten recovered recombinants (#1–10) from the dead donor assay (donor W83, recipient 33277) were tested for the presence of the donor fimA type IV allele using PCR amplification. In addition, to determine if regions further downstream of fimA were also swapped, the recombinants were tested for the presence of donor fimC, fimD , and fimE . ‘ X ’ indicates the presence of the donor gene and ‘ O ’ indicates the presence of the recipient gene in the tested strains.

    Journal: PLoS ONE

    Article Title: Genetic Exchange of Fimbrial Alleles Exemplifies the Adaptive Virulence Strategy of Porphyromonas gingivalis

    doi: 10.1371/journal.pone.0091696

    Figure Lengend Snippet: fim region able to swap during natural competence despite limited conservation regions. (A.) Complete fim gene region ( fimA-fimE ) and flanking genes are shown for sequenced P. gingivalis strains 33277 fimA (I ), TDC60 fimA (II ), and W83 fimA (IV ). Overall sequence conservation between all strains is displayed. Green indicates the most conservation while yellow, orange, and finally red signifies the least DNA conservation between strains. Distances are shown in base pairs. Figure modified from CLC Genomic Workbench 6, accession numbers 33277(NC_010729), TD60 (NC_015571), and W83 (NC_002950). (B.) Chromosomal preparations from a random sample of ten recovered recombinants (#1–10) from the dead donor assay (donor W83, recipient 33277) were tested for the presence of the donor fimA type IV allele using PCR amplification. In addition, to determine if regions further downstream of fimA were also swapped, the recombinants were tested for the presence of donor fimC, fimD , and fimE . ‘ X ’ indicates the presence of the donor gene and ‘ O ’ indicates the presence of the recipient gene in the tested strains.

    Article Snippet: RNA Isolation Cells of P. gingivalis were washed and resuspended in pre-reduced 1X phosphate buffered saline (PBS) to a final concentration of 1×108 CFU/ml.

    Techniques: Sequencing, Modification, Polymerase Chain Reaction, Amplification

    Invasion of human gingival tissue by fimA isogenic exchange mutants. A 24-hour invasion of gingival fibroblast cells by P. gingivalis was quantitated by an antibiotic protection assay. There is no significant difference in invasion rates between A1 and 33277; YPF1 has a reduced invasion efficiency. Introduction of the type III or type IV fimA alleles into 33277 leads to enhanced invasion capability compared to the A1 strain. All data are presented as the mean +/− SD from five independent trials performed in triplicate. Statistically significant differences calculated between A1:A3 *, A1:A4 **, and A1:YPF1 *** (p

    Journal: PLoS ONE

    Article Title: Genetic Exchange of Fimbrial Alleles Exemplifies the Adaptive Virulence Strategy of Porphyromonas gingivalis

    doi: 10.1371/journal.pone.0091696

    Figure Lengend Snippet: Invasion of human gingival tissue by fimA isogenic exchange mutants. A 24-hour invasion of gingival fibroblast cells by P. gingivalis was quantitated by an antibiotic protection assay. There is no significant difference in invasion rates between A1 and 33277; YPF1 has a reduced invasion efficiency. Introduction of the type III or type IV fimA alleles into 33277 leads to enhanced invasion capability compared to the A1 strain. All data are presented as the mean +/− SD from five independent trials performed in triplicate. Statistically significant differences calculated between A1:A3 *, A1:A4 **, and A1:YPF1 *** (p

    Article Snippet: RNA Isolation Cells of P. gingivalis were washed and resuspended in pre-reduced 1X phosphate buffered saline (PBS) to a final concentration of 1×108 CFU/ml.

    Techniques:

    fimA allele types are actively exchanged via natural competence. (A.) Genomic constructs for the donor strains 33277 (top) and W83 (bottom). Donor strains contained the genomic insertion ermF (green) directly downstream of fimA type I (yellow) or IV (blue). Open reading frame sizes are proportional; created using CLC Genomic Workbench 6, from sequence accession numbers NC_010729 (33277) and NC_002950 (W83). (B.) The DNA uptake efficiencies of experiments with four live recipient P. gingivalis strains, using either the 33277 or W83 dead donor strain. fimA allele transfer frequencies are calculated as the number of recovered recombinants (Rif r , Erm r ) divided by the number of input recipient cells. All data are presented as the mean +/− standard deviation (SD) from a minimum of three independent trials performed in triplicate. All recipient strains were rifampin resistant. Statistical comparisons between all groups were done by one-way Anova. (C.) DNA sequence alignment of the fimA recipient alleles from figure 1B . Degree of conservation between the four sequences is shown, with a scale from green to red representing 100% to 0% conservation. Regions responsible for genetic recombination are underlined by dashed lines, and are > 98% identical between any donor and recipient DNA sequence. DNA sequences from strain 53977 and 49417 are available at GenBank accession numbers KF770042 and KF770043.

    Journal: PLoS ONE

    Article Title: Genetic Exchange of Fimbrial Alleles Exemplifies the Adaptive Virulence Strategy of Porphyromonas gingivalis

    doi: 10.1371/journal.pone.0091696

    Figure Lengend Snippet: fimA allele types are actively exchanged via natural competence. (A.) Genomic constructs for the donor strains 33277 (top) and W83 (bottom). Donor strains contained the genomic insertion ermF (green) directly downstream of fimA type I (yellow) or IV (blue). Open reading frame sizes are proportional; created using CLC Genomic Workbench 6, from sequence accession numbers NC_010729 (33277) and NC_002950 (W83). (B.) The DNA uptake efficiencies of experiments with four live recipient P. gingivalis strains, using either the 33277 or W83 dead donor strain. fimA allele transfer frequencies are calculated as the number of recovered recombinants (Rif r , Erm r ) divided by the number of input recipient cells. All data are presented as the mean +/− standard deviation (SD) from a minimum of three independent trials performed in triplicate. All recipient strains were rifampin resistant. Statistical comparisons between all groups were done by one-way Anova. (C.) DNA sequence alignment of the fimA recipient alleles from figure 1B . Degree of conservation between the four sequences is shown, with a scale from green to red representing 100% to 0% conservation. Regions responsible for genetic recombination are underlined by dashed lines, and are > 98% identical between any donor and recipient DNA sequence. DNA sequences from strain 53977 and 49417 are available at GenBank accession numbers KF770042 and KF770043.

    Article Snippet: RNA Isolation Cells of P. gingivalis were washed and resuspended in pre-reduced 1X phosphate buffered saline (PBS) to a final concentration of 1×108 CFU/ml.

    Techniques: Construct, Sequencing, Standard Deviation

    Electron micrographs of P. gingivalis surface structures. (A.) P. gingivalis strains 33277 A1 and 33277 (both fimA type I). (B.) P. gingivalis 33277 A3 and 49417 (both fimA type III). (C.) P. gingivalis 33277 A4 and W83 (both fimA type IV). (D.) P. gingivalis strain 33277 fimA minus, YPF1. Samples were stained with 2% PTA and subjected to transmission electron microscopy, viewed using a JEOL JEM 1010 microscope. Bars = 500 nm. Black box indicates area at 50,000x direct magnification (50 k) that was examined further at 100,000x direct magnification (100 k).

    Journal: PLoS ONE

    Article Title: Genetic Exchange of Fimbrial Alleles Exemplifies the Adaptive Virulence Strategy of Porphyromonas gingivalis

    doi: 10.1371/journal.pone.0091696

    Figure Lengend Snippet: Electron micrographs of P. gingivalis surface structures. (A.) P. gingivalis strains 33277 A1 and 33277 (both fimA type I). (B.) P. gingivalis 33277 A3 and 49417 (both fimA type III). (C.) P. gingivalis 33277 A4 and W83 (both fimA type IV). (D.) P. gingivalis strain 33277 fimA minus, YPF1. Samples were stained with 2% PTA and subjected to transmission electron microscopy, viewed using a JEOL JEM 1010 microscope. Bars = 500 nm. Black box indicates area at 50,000x direct magnification (50 k) that was examined further at 100,000x direct magnification (100 k).

    Article Snippet: RNA Isolation Cells of P. gingivalis were washed and resuspended in pre-reduced 1X phosphate buffered saline (PBS) to a final concentration of 1×108 CFU/ml.

    Techniques: Staining, Transmission Assay, Electron Microscopy, Microscopy

    Bioinformatics analysis of P. gingivalis FimA protein alleles. (A.) Predicted proteins motifs are shown for FimA type I, III and IV, from P. gingivalis strains 33277 (I), 49417 (III), and W83 (IV), respectively. Protein motif analysis and alignment were performed with CLC Genomics Workbench software version 6. (B.) Alignment of the N-terminal regions of fimbrial proteins. Boxed sequences (MTAC) are the lipoprotein sorting signals. Black arrows indicate the predicted gingipain cleavage sites of the leader peptide. Residue conservation between all strains is displayed as a color bar below the alignment. Red highlighting of individual residues shows deviation from the aligned consensus. (C.) Guide to color and symbols in A and B. The pilus biogenesis symbol represents a match to the pfam Pilus biogenesis CpaD protein (pilus_cpaD) Accession: PF09476. The aminotransferase represents a match to pfam Aminotransferase class I and II, Accession: PF00155. The trypsin symbol represents a match to pfam Trypsin, Accession: PF00089. Sequence conservation between all strains is displayed below the alignment. Green indicates the most conservation while red signifies the least conservation between strains.

    Journal: PLoS ONE

    Article Title: Genetic Exchange of Fimbrial Alleles Exemplifies the Adaptive Virulence Strategy of Porphyromonas gingivalis

    doi: 10.1371/journal.pone.0091696

    Figure Lengend Snippet: Bioinformatics analysis of P. gingivalis FimA protein alleles. (A.) Predicted proteins motifs are shown for FimA type I, III and IV, from P. gingivalis strains 33277 (I), 49417 (III), and W83 (IV), respectively. Protein motif analysis and alignment were performed with CLC Genomics Workbench software version 6. (B.) Alignment of the N-terminal regions of fimbrial proteins. Boxed sequences (MTAC) are the lipoprotein sorting signals. Black arrows indicate the predicted gingipain cleavage sites of the leader peptide. Residue conservation between all strains is displayed as a color bar below the alignment. Red highlighting of individual residues shows deviation from the aligned consensus. (C.) Guide to color and symbols in A and B. The pilus biogenesis symbol represents a match to the pfam Pilus biogenesis CpaD protein (pilus_cpaD) Accession: PF09476. The aminotransferase represents a match to pfam Aminotransferase class I and II, Accession: PF00155. The trypsin symbol represents a match to pfam Trypsin, Accession: PF00089. Sequence conservation between all strains is displayed below the alignment. Green indicates the most conservation while red signifies the least conservation between strains.

    Article Snippet: RNA Isolation Cells of P. gingivalis were washed and resuspended in pre-reduced 1X phosphate buffered saline (PBS) to a final concentration of 1×108 CFU/ml.

    Techniques: Software, Sequencing

    Transmission electron micrographs demonstrating P. gingivalis invasion of EC. (A and B) BAEC with P. gingivalis A7436. (A) At the cell surface, bacteria appear to induce EC structural rearrangements consistent with an endocytic mechanism. (B) Internalized bacteria are found within vacuole. (C) BAEC with P. gingivalis 381. Note the apparent contact between microfilamentous cellular components and surface-adhering P. gingivalis . (D) BAEC with P. gingivalis fimA mutant DPG3. Absence of intimate interaction between EC surface and bacteria. (E and F) FBHEC incubated with P. gingivalis A7436. Surface adherence (E) and engulfment in vacuole (F). Arrows in all panels point to P. gingivalis . Bars on each image are 0.5 μm unless otherwise specified. Composite image was constructed with Adobe Photoshop 3.0.

    Journal: Infection and Immunity

    Article Title: Invasion of Aortic and Heart Endothelial Cells by Porphyromonas gingivalis

    doi:

    Figure Lengend Snippet: Transmission electron micrographs demonstrating P. gingivalis invasion of EC. (A and B) BAEC with P. gingivalis A7436. (A) At the cell surface, bacteria appear to induce EC structural rearrangements consistent with an endocytic mechanism. (B) Internalized bacteria are found within vacuole. (C) BAEC with P. gingivalis 381. Note the apparent contact between microfilamentous cellular components and surface-adhering P. gingivalis . (D) BAEC with P. gingivalis fimA mutant DPG3. Absence of intimate interaction between EC surface and bacteria. (E and F) FBHEC incubated with P. gingivalis A7436. Surface adherence (E) and engulfment in vacuole (F). Arrows in all panels point to P. gingivalis . Bars on each image are 0.5 μm unless otherwise specified. Composite image was constructed with Adobe Photoshop 3.0.

    Article Snippet: It appears that the receptors required for adherence of P. gingivalis may be present on the EC surface since de novo protein synthesis of EC is not required for invasion.

    Techniques: Transmission Assay, Mutagenesis, Incubation, Construct

    The effects of S. mitis , E. coli , heat-killed P. gingivalis and the supernatant prepared from P. gingivalis cultures on the microglial response. ( a–l ) The dynamic response of the microglial processes to the local injection of various bacteria (3.6 × 10 5 CFU ml −1 ) at ZT2. ( a–c ) S. mitis (S.m), ( d–f ) E. coli (E.c), ( g–i ) heat killed P.g (HK-P.g; 95 °C, 10 min), ( j–l ) P.g culture supernatant (P.g SN). Scale bar: 10 μm. ( m ) The kinetics of the mean fluorescent change in the microglial response measured at 30 min after the injection of various bacteria. The data are presented as the mean ± S.E.M. (N = 3 mice, n = 4–5 each). A one-way ANOVA with a post hoc Tukey’s test; P.g vs S.m: p > 0.9999, Pg. vs. E.c: p = 0.9953, P.g vs. HK-P.g: p = 0.4244, S.m vs. E.c: p = 0.9939, S.m vs. HK-P.g: p = 0.4126, E.c vs. HK-P.g: p = 0.5521. ( n ) The kinetics of the mean fluorescent change in the microglial response measured at 30 min after the injection of P. gingivalis . The data are presented as the mean ± S.E.M. (N = 3 mice, n = 5–6 each). A two-tailed unpaired t -test; P.g vs BHI (P.g culture medium, as a control): p = 0.0856.

    Journal: Scientific Reports

    Article Title: Diurnal dynamic behavior of microglia in response to infected bacteria through the UDP-P2Y6 receptor system

    doi: 10.1038/srep30006

    Figure Lengend Snippet: The effects of S. mitis , E. coli , heat-killed P. gingivalis and the supernatant prepared from P. gingivalis cultures on the microglial response. ( a–l ) The dynamic response of the microglial processes to the local injection of various bacteria (3.6 × 10 5 CFU ml −1 ) at ZT2. ( a–c ) S. mitis (S.m), ( d–f ) E. coli (E.c), ( g–i ) heat killed P.g (HK-P.g; 95 °C, 10 min), ( j–l ) P.g culture supernatant (P.g SN). Scale bar: 10 μm. ( m ) The kinetics of the mean fluorescent change in the microglial response measured at 30 min after the injection of various bacteria. The data are presented as the mean ± S.E.M. (N = 3 mice, n = 4–5 each). A one-way ANOVA with a post hoc Tukey’s test; P.g vs S.m: p > 0.9999, Pg. vs. E.c: p = 0.9953, P.g vs. HK-P.g: p = 0.4244, S.m vs. E.c: p = 0.9939, S.m vs. HK-P.g: p = 0.4126, E.c vs. HK-P.g: p = 0.5521. ( n ) The kinetics of the mean fluorescent change in the microglial response measured at 30 min after the injection of P. gingivalis . The data are presented as the mean ± S.E.M. (N = 3 mice, n = 5–6 each). A two-tailed unpaired t -test; P.g vs BHI (P.g culture medium, as a control): p = 0.0856.

    Article Snippet: P. gingivalis was maintained on blood agar plates and grown in enriched BHI broth (containing, per liter, 37 g of brain heart infusion, 5 g of yeast extract, 1 g of cysteine, 5 mg of hemin, and 1 mg of vitamin K1) under anaerobic conditions (10% CO2 , 10% H2 , 80% N2 ) .

    Techniques: Injection, Mouse Assay, Two Tailed Test

    The differential diurnal variation in the dynamic behavior of the microglial processes in response to the focal injection of ATP and P. gingivalis. ( a–l ) The dynamic response of the microglial processes to the local injection of 10 mM ATP ( a–f ) and 3.6 × 10 5 CFU ml −1 P. gingivalis ( g–l ) in an open-skull preparation of CX3CR1 GFP /+ mice at ZT2 and ZT14. Scale bar: 10 μm. ( m,n ) The kinetics of the mean fluorescent change in the microglial response to ATP ( m ) and P. gingivalis ( n ) at ZT2 and ZT14. The data are presented as the mean ± S.E.M. (N = 3 mice, n = 3–6 each). A two-way repeated measure ANOVA with Sidak’s test (ZT2 versus ZT14); from 10 to 40 min: p = 0.9985, p = 0.6209, p = 0.0030, p = 0.0006, p = 0.0082, p = 0.0939, p = 0.0455 ( m ), p = 0.0030, p = 0.0002, p = 0.0001; from 25 to 40 min: p = 0.0001 ( n – p ) The maximum distance from the reactive microglia to the injection of ATP ( o ) and P. gingivalis (P.g) ( p ) at ZT2 and ZT14. The data are presented as the mean ± S.E.M. A two-tailed unpaired t -test; ** p = 0.0042 ( o ), * p = 0.0173 ( p ).

    Journal: Scientific Reports

    Article Title: Diurnal dynamic behavior of microglia in response to infected bacteria through the UDP-P2Y6 receptor system

    doi: 10.1038/srep30006

    Figure Lengend Snippet: The differential diurnal variation in the dynamic behavior of the microglial processes in response to the focal injection of ATP and P. gingivalis. ( a–l ) The dynamic response of the microglial processes to the local injection of 10 mM ATP ( a–f ) and 3.6 × 10 5 CFU ml −1 P. gingivalis ( g–l ) in an open-skull preparation of CX3CR1 GFP /+ mice at ZT2 and ZT14. Scale bar: 10 μm. ( m,n ) The kinetics of the mean fluorescent change in the microglial response to ATP ( m ) and P. gingivalis ( n ) at ZT2 and ZT14. The data are presented as the mean ± S.E.M. (N = 3 mice, n = 3–6 each). A two-way repeated measure ANOVA with Sidak’s test (ZT2 versus ZT14); from 10 to 40 min: p = 0.9985, p = 0.6209, p = 0.0030, p = 0.0006, p = 0.0082, p = 0.0939, p = 0.0455 ( m ), p = 0.0030, p = 0.0002, p = 0.0001; from 25 to 40 min: p = 0.0001 ( n – p ) The maximum distance from the reactive microglia to the injection of ATP ( o ) and P. gingivalis (P.g) ( p ) at ZT2 and ZT14. The data are presented as the mean ± S.E.M. A two-tailed unpaired t -test; ** p = 0.0042 ( o ), * p = 0.0173 ( p ).

    Article Snippet: P. gingivalis was maintained on blood agar plates and grown in enriched BHI broth (containing, per liter, 37 g of brain heart infusion, 5 g of yeast extract, 1 g of cysteine, 5 mg of hemin, and 1 mg of vitamin K1) under anaerobic conditions (10% CO2 , 10% H2 , 80% N2 ) .

    Techniques: Injection, Mouse Assay, Two Tailed Test

    The possible involvement of UDP in the microglial response to bacterial infection. ( a–c ) The dynamic response of the microglial processes to the local injection of 10 mM UDP and P. gingivalis in the presence of 1 μM MRS2578 ( d–f ) at ZT2. Scale bar: 10 μm. ( g ) The kinetics of the mean fluorescent change in the microglial response measured at the 30 min after the injection of UDP and P. gingivalis in the presence of MRS2578. The data are presented as the mean ± S.E.M. (N = 3 mice, n = 5–6 each). A one-way ANOVA with Dunnett’s test; P.g vs. UDP: p = 0.2162, P.g vs. MRS3578: * p = 0.0277. ( h ) MG6 microglial cells released UDP after infection with bacteria at indicated MOI. P.g, P. gingivalis ; S.m, S. mitis ; E.c, E. coli . The data are presented as the mean ± S.E.M. (n = 4 each). A one-way ANOVA with Dunnett’s test as compared with the control; P.g 1:5, p = 0.0001; P.g 1:10, p = 0.0001; S.m 1:5, p = 0.0021; S.m 1:10, p = 0.0001; E.c 1:5, p = 0.0049; E.c 1:10, p = 0.0001.

    Journal: Scientific Reports

    Article Title: Diurnal dynamic behavior of microglia in response to infected bacteria through the UDP-P2Y6 receptor system

    doi: 10.1038/srep30006

    Figure Lengend Snippet: The possible involvement of UDP in the microglial response to bacterial infection. ( a–c ) The dynamic response of the microglial processes to the local injection of 10 mM UDP and P. gingivalis in the presence of 1 μM MRS2578 ( d–f ) at ZT2. Scale bar: 10 μm. ( g ) The kinetics of the mean fluorescent change in the microglial response measured at the 30 min after the injection of UDP and P. gingivalis in the presence of MRS2578. The data are presented as the mean ± S.E.M. (N = 3 mice, n = 5–6 each). A one-way ANOVA with Dunnett’s test; P.g vs. UDP: p = 0.2162, P.g vs. MRS3578: * p = 0.0277. ( h ) MG6 microglial cells released UDP after infection with bacteria at indicated MOI. P.g, P. gingivalis ; S.m, S. mitis ; E.c, E. coli . The data are presented as the mean ± S.E.M. (n = 4 each). A one-way ANOVA with Dunnett’s test as compared with the control; P.g 1:5, p = 0.0001; P.g 1:10, p = 0.0001; S.m 1:5, p = 0.0021; S.m 1:10, p = 0.0001; E.c 1:5, p = 0.0049; E.c 1:10, p = 0.0001.

    Article Snippet: P. gingivalis was maintained on blood agar plates and grown in enriched BHI broth (containing, per liter, 37 g of brain heart infusion, 5 g of yeast extract, 1 g of cysteine, 5 mg of hemin, and 1 mg of vitamin K1) under anaerobic conditions (10% CO2 , 10% H2 , 80% N2 ) .

    Techniques: Infection, Injection, Mouse Assay

    The possible involvement of extracellular nucleotides in the microglial response to bacterial infection. ( a–i ) The dynamic response of microglial processes to the local injection of P. gingivalis in the presence of various compounds at ZT2. ( a–c ) 5 U ml −1 apyrase, ( d–f ) 1 μM PSB. Scale bar: 10 μm. ( g ) The kinetics of the mean fluorescent change in the microglial response measured at 30 min after the injection of P. gingivalis in the presence of various compounds at ZT2. The data are presented as the mean ± S.E.M. (N = 3 mice, n = 3–4 each). A one-way ANOVA with Dunnett’s test; P.g vs. apyrase: * p = 0.0230; P.g vs. PSB: p = 0.4400.

    Journal: Scientific Reports

    Article Title: Diurnal dynamic behavior of microglia in response to infected bacteria through the UDP-P2Y6 receptor system

    doi: 10.1038/srep30006

    Figure Lengend Snippet: The possible involvement of extracellular nucleotides in the microglial response to bacterial infection. ( a–i ) The dynamic response of microglial processes to the local injection of P. gingivalis in the presence of various compounds at ZT2. ( a–c ) 5 U ml −1 apyrase, ( d–f ) 1 μM PSB. Scale bar: 10 μm. ( g ) The kinetics of the mean fluorescent change in the microglial response measured at 30 min after the injection of P. gingivalis in the presence of various compounds at ZT2. The data are presented as the mean ± S.E.M. (N = 3 mice, n = 3–4 each). A one-way ANOVA with Dunnett’s test; P.g vs. apyrase: * p = 0.0230; P.g vs. PSB: p = 0.4400.

    Article Snippet: P. gingivalis was maintained on blood agar plates and grown in enriched BHI broth (containing, per liter, 37 g of brain heart infusion, 5 g of yeast extract, 1 g of cysteine, 5 mg of hemin, and 1 mg of vitamin K1) under anaerobic conditions (10% CO2 , 10% H2 , 80% N2 ) .

    Techniques: Infection, Injection, Mouse Assay

    P2Y 6 R knockdown inhibited the dynamic behavior of the microglial processes in response to bacterial injection. ( a–c ) The determination of the transfection efficiency using BLOCK-iT Alexa Fluor Red Fluorescent Control. ( a ) The BLOCK-iT Alexa Fluor Red Fluorescent Control (20 pmol) was applied through a small craniotomy. ( b ) CLSM images showed the intracellular uptake of the BLOCK-iT Fluorescent Oligo (Red) in the somatosensory cortex at 24 h after transfection. ( c ) CLSM images merged with the BLOCK-iT Fluorescent Oligo (red) and microglia (green) in the somatosensory cortex of CX3CR1 +/ GFP mice at 24 h after transfection. Scale bar: 10 μm. ( d–i ) The dynamic response of the microglial processes to the local injection of P. gingivalis after the administration of the control siRNA ( d–f ) and P2Y 6 R siRNA. Scale bar: 10 μm. ( j ) The kinetics of the mean fluorescent change in the microglial response measured at 30 min after the injection of P. gingivalis following the administration of the control siRNA (siCont) and P2Y 6 R siRNA (siRNA). The data are presented as the mean ± S.E.M. (N = 3 mice, n = 5 each). A two-tailed unpaired t -test; ** p = 0.0051. ( k ) The relative P2ry6 mRNA expression level after the administration of the control siRNA and P2Y 6 R siRNA. The data are presented as the mean ± S.E.M. (N = 3 animal, n = 14 each). A two-tailed unpaired t -test; * p = 0.0373.

    Journal: Scientific Reports

    Article Title: Diurnal dynamic behavior of microglia in response to infected bacteria through the UDP-P2Y6 receptor system

    doi: 10.1038/srep30006

    Figure Lengend Snippet: P2Y 6 R knockdown inhibited the dynamic behavior of the microglial processes in response to bacterial injection. ( a–c ) The determination of the transfection efficiency using BLOCK-iT Alexa Fluor Red Fluorescent Control. ( a ) The BLOCK-iT Alexa Fluor Red Fluorescent Control (20 pmol) was applied through a small craniotomy. ( b ) CLSM images showed the intracellular uptake of the BLOCK-iT Fluorescent Oligo (Red) in the somatosensory cortex at 24 h after transfection. ( c ) CLSM images merged with the BLOCK-iT Fluorescent Oligo (red) and microglia (green) in the somatosensory cortex of CX3CR1 +/ GFP mice at 24 h after transfection. Scale bar: 10 μm. ( d–i ) The dynamic response of the microglial processes to the local injection of P. gingivalis after the administration of the control siRNA ( d–f ) and P2Y 6 R siRNA. Scale bar: 10 μm. ( j ) The kinetics of the mean fluorescent change in the microglial response measured at 30 min after the injection of P. gingivalis following the administration of the control siRNA (siCont) and P2Y 6 R siRNA (siRNA). The data are presented as the mean ± S.E.M. (N = 3 mice, n = 5 each). A two-tailed unpaired t -test; ** p = 0.0051. ( k ) The relative P2ry6 mRNA expression level after the administration of the control siRNA and P2Y 6 R siRNA. The data are presented as the mean ± S.E.M. (N = 3 animal, n = 14 each). A two-tailed unpaired t -test; * p = 0.0373.

    Article Snippet: P. gingivalis was maintained on blood agar plates and grown in enriched BHI broth (containing, per liter, 37 g of brain heart infusion, 5 g of yeast extract, 1 g of cysteine, 5 mg of hemin, and 1 mg of vitamin K1) under anaerobic conditions (10% CO2 , 10% H2 , 80% N2 ) .

    Techniques: Injection, Transfection, Blocking Assay, Confocal Laser Scanning Microscopy, Mouse Assay, Two Tailed Test, Expressing

    Effects of OMV preexposure on TNF secretion in response to secondary stimulation with live P. gingivalis . (A) Human monocytes were prestimulated or not prestimulated with OMV for 24 h; after washing, cells were left unstimulated (medium) or restimulated

    Journal: Infection and Immunity

    Article Title: Porphyromonas gingivalis Outer Membrane Vesicles Induce Selective Tumor Necrosis Factor Tolerance in a Toll-Like Receptor 4- and mTOR-Dependent Manner

    doi: 10.1128/IAI.01390-15

    Figure Lengend Snippet: Effects of OMV preexposure on TNF secretion in response to secondary stimulation with live P. gingivalis . (A) Human monocytes were prestimulated or not prestimulated with OMV for 24 h; after washing, cells were left unstimulated (medium) or restimulated

    Article Snippet: DNA from the Staphylococcus aureus SA113 Δ lgt strain (devoid of TLR2 activity) ( ) or P. gingivalis and poly(dA·dT) (Sigma-Aldrich) (100 ng/well) were transfected with Lipofectamine (Invitrogen) (100 ng DNA per well in Opti-MEM [Gibco/Life Science]).

    Techniques:

    Role of fimbriae, gingipains, and endocytosis in TNF tolerance. (A) Monocytes were left unstimulated or prestimulated with Salmonella Minnesota LPS, P. gingivalis (ATCC 33277) OMV, or OMV of the nonfimbriated P. gingivalis W83 strain for 24 h. Cells were

    Journal: Infection and Immunity

    Article Title: Porphyromonas gingivalis Outer Membrane Vesicles Induce Selective Tumor Necrosis Factor Tolerance in a Toll-Like Receptor 4- and mTOR-Dependent Manner

    doi: 10.1128/IAI.01390-15

    Figure Lengend Snippet: Role of fimbriae, gingipains, and endocytosis in TNF tolerance. (A) Monocytes were left unstimulated or prestimulated with Salmonella Minnesota LPS, P. gingivalis (ATCC 33277) OMV, or OMV of the nonfimbriated P. gingivalis W83 strain for 24 h. Cells were

    Article Snippet: DNA from the Staphylococcus aureus SA113 Δ lgt strain (devoid of TLR2 activity) ( ) or P. gingivalis and poly(dA·dT) (Sigma-Aldrich) (100 ng/well) were transfected with Lipofectamine (Invitrogen) (100 ng DNA per well in Opti-MEM [Gibco/Life Science]).

    Techniques:

    Secretion of proinflammatory cytokines in response to P. gingivalis OMV. (A) Trans-electron microscopy of the purified OMV. The image shows a negative stain with 2% methylamine tungstate and is representative of the results of n = 6 experiments. (B and

    Journal: Infection and Immunity

    Article Title: Porphyromonas gingivalis Outer Membrane Vesicles Induce Selective Tumor Necrosis Factor Tolerance in a Toll-Like Receptor 4- and mTOR-Dependent Manner

    doi: 10.1128/IAI.01390-15

    Figure Lengend Snippet: Secretion of proinflammatory cytokines in response to P. gingivalis OMV. (A) Trans-electron microscopy of the purified OMV. The image shows a negative stain with 2% methylamine tungstate and is representative of the results of n = 6 experiments. (B and

    Article Snippet: DNA from the Staphylococcus aureus SA113 Δ lgt strain (devoid of TLR2 activity) ( ) or P. gingivalis and poly(dA·dT) (Sigma-Aldrich) (100 ng/well) were transfected with Lipofectamine (Invitrogen) (100 ng DNA per well in Opti-MEM [Gibco/Life Science]).

    Techniques: Electron Microscopy, Purification, Staining

    Role of TLR2 and TLR4 in OMV-mediated abrogation of the TNF response. (A to D) Human monocytes were prestimulated with OMV or not prestimulated; live P. gingivalis was used for secondary challenge after washing. The experiments were performed in the presence

    Journal: Infection and Immunity

    Article Title: Porphyromonas gingivalis Outer Membrane Vesicles Induce Selective Tumor Necrosis Factor Tolerance in a Toll-Like Receptor 4- and mTOR-Dependent Manner

    doi: 10.1128/IAI.01390-15

    Figure Lengend Snippet: Role of TLR2 and TLR4 in OMV-mediated abrogation of the TNF response. (A to D) Human monocytes were prestimulated with OMV or not prestimulated; live P. gingivalis was used for secondary challenge after washing. The experiments were performed in the presence

    Article Snippet: DNA from the Staphylococcus aureus SA113 Δ lgt strain (devoid of TLR2 activity) ( ) or P. gingivalis and poly(dA·dT) (Sigma-Aldrich) (100 ng/well) were transfected with Lipofectamine (Invitrogen) (100 ng DNA per well in Opti-MEM [Gibco/Life Science]).

    Techniques:

    LXA 4 inhibits P. gingivalis -induced upregulation of CD11b/CD18 and exposure of the CD11b/CD18 high-affinity epitope on neutrophils. Whole blood was incubated in the presence or absence of LXA 4 (100 or 500 nM) for 15 min and subsequently stimulated with

    Journal: Infection and Immunity

    Article Title: Lipoxin A4 Inhibits Porphyromonas gingivalis-Induced Aggregation and Reactive Oxygen Species Production by Modulating Neutrophil-Platelet Interaction and CD11b Expression ▿-Induced Aggregation and Reactive Oxygen Species Production by Modulating Neutrophil-Platelet Interaction and CD11b Expression ▿ †-Induced Aggregation and Reactive Oxygen Species Production by Modulating Neutrophil-Platelet Interaction and CD11b Expression ▿ † ‡

    doi: 10.1128/IAI.00777-10

    Figure Lengend Snippet: LXA 4 inhibits P. gingivalis -induced upregulation of CD11b/CD18 and exposure of the CD11b/CD18 high-affinity epitope on neutrophils. Whole blood was incubated in the presence or absence of LXA 4 (100 or 500 nM) for 15 min and subsequently stimulated with

    Article Snippet: In short, whole blood was incubated at 37°C in the presence or absence of LXA4 (1, 100, 250, or 500 nM) for 15 min and subsequently stimulated with P. gingivalis (1 × 107 CFU/ml blood) for 10 min. To detect the total CD11b expression, an R-phycoerythrin (RPE)-conjugated anti-human CD11b antibody (mouse monoclonal antibody; clone 2LPM19c; Dako, Glostrup, Denmark) was added 5 min after the onset of bacterial stimulation.

    Techniques: Incubation

    Effect of LXA 4 on P. gingivalis -induced aggregation and ROS production in platelet-depleted whole blood. Platelet-depleted whole blood was incubated in the absence or presence of LXA 4 (500 nM). Aggregation was measured as changes in impedance and ROS

    Journal: Infection and Immunity

    Article Title: Lipoxin A4 Inhibits Porphyromonas gingivalis-Induced Aggregation and Reactive Oxygen Species Production by Modulating Neutrophil-Platelet Interaction and CD11b Expression ▿-Induced Aggregation and Reactive Oxygen Species Production by Modulating Neutrophil-Platelet Interaction and CD11b Expression ▿ †-Induced Aggregation and Reactive Oxygen Species Production by Modulating Neutrophil-Platelet Interaction and CD11b Expression ▿ † ‡

    doi: 10.1128/IAI.00777-10

    Figure Lengend Snippet: Effect of LXA 4 on P. gingivalis -induced aggregation and ROS production in platelet-depleted whole blood. Platelet-depleted whole blood was incubated in the absence or presence of LXA 4 (500 nM). Aggregation was measured as changes in impedance and ROS

    Article Snippet: In short, whole blood was incubated at 37°C in the presence or absence of LXA4 (1, 100, 250, or 500 nM) for 15 min and subsequently stimulated with P. gingivalis (1 × 107 CFU/ml blood) for 10 min. To detect the total CD11b expression, an R-phycoerythrin (RPE)-conjugated anti-human CD11b antibody (mouse monoclonal antibody; clone 2LPM19c; Dako, Glostrup, Denmark) was added 5 min after the onset of bacterial stimulation.

    Techniques: Incubation

    LXA 4 inhibits P. gingivalis -induced formation of cellular aggregates. Whole blood was incubated in the absence or presence of 500 nM LXA 4 for 15 min before being stimulated with P. gingivalis (1 × 10 7 CFU/ml blood) for 25 min. An aliquot of each

    Journal: Infection and Immunity

    Article Title: Lipoxin A4 Inhibits Porphyromonas gingivalis-Induced Aggregation and Reactive Oxygen Species Production by Modulating Neutrophil-Platelet Interaction and CD11b Expression ▿-Induced Aggregation and Reactive Oxygen Species Production by Modulating Neutrophil-Platelet Interaction and CD11b Expression ▿ †-Induced Aggregation and Reactive Oxygen Species Production by Modulating Neutrophil-Platelet Interaction and CD11b Expression ▿ † ‡

    doi: 10.1128/IAI.00777-10

    Figure Lengend Snippet: LXA 4 inhibits P. gingivalis -induced formation of cellular aggregates. Whole blood was incubated in the absence or presence of 500 nM LXA 4 for 15 min before being stimulated with P. gingivalis (1 × 10 7 CFU/ml blood) for 25 min. An aliquot of each

    Article Snippet: In short, whole blood was incubated at 37°C in the presence or absence of LXA4 (1, 100, 250, or 500 nM) for 15 min and subsequently stimulated with P. gingivalis (1 × 107 CFU/ml blood) for 10 min. To detect the total CD11b expression, an R-phycoerythrin (RPE)-conjugated anti-human CD11b antibody (mouse monoclonal antibody; clone 2LPM19c; Dako, Glostrup, Denmark) was added 5 min after the onset of bacterial stimulation.

    Techniques: Incubation

    LXA 4 inhibits P. gingivalis -induced aggregation and ROS production in whole blood. Whole blood was incubated for 15 min in the absence or presence of lipoxin A 4 (LX; 1 to 500 nM) prior to stimulation with P. gingivalis (P.g; 1 × 10 7 CFU/ml blood)

    Journal: Infection and Immunity

    Article Title: Lipoxin A4 Inhibits Porphyromonas gingivalis-Induced Aggregation and Reactive Oxygen Species Production by Modulating Neutrophil-Platelet Interaction and CD11b Expression ▿-Induced Aggregation and Reactive Oxygen Species Production by Modulating Neutrophil-Platelet Interaction and CD11b Expression ▿ †-Induced Aggregation and Reactive Oxygen Species Production by Modulating Neutrophil-Platelet Interaction and CD11b Expression ▿ † ‡

    doi: 10.1128/IAI.00777-10

    Figure Lengend Snippet: LXA 4 inhibits P. gingivalis -induced aggregation and ROS production in whole blood. Whole blood was incubated for 15 min in the absence or presence of lipoxin A 4 (LX; 1 to 500 nM) prior to stimulation with P. gingivalis (P.g; 1 × 10 7 CFU/ml blood)

    Article Snippet: In short, whole blood was incubated at 37°C in the presence or absence of LXA4 (1, 100, 250, or 500 nM) for 15 min and subsequently stimulated with P. gingivalis (1 × 107 CFU/ml blood) for 10 min. To detect the total CD11b expression, an R-phycoerythrin (RPE)-conjugated anti-human CD11b antibody (mouse monoclonal antibody; clone 2LPM19c; Dako, Glostrup, Denmark) was added 5 min after the onset of bacterial stimulation.

    Techniques: Incubation

    LXA 4 does not affect P. gingivalis -induced ROS production in isolated neutrophils. Isolated polymorphonuclear cells (PMN), consisting predominantly of neutrophils, were preincubated for 15 min at 37°C in the presence or absence of LXA 4 (1 or 100

    Journal: Infection and Immunity

    Article Title: Lipoxin A4 Inhibits Porphyromonas gingivalis-Induced Aggregation and Reactive Oxygen Species Production by Modulating Neutrophil-Platelet Interaction and CD11b Expression ▿-Induced Aggregation and Reactive Oxygen Species Production by Modulating Neutrophil-Platelet Interaction and CD11b Expression ▿ †-Induced Aggregation and Reactive Oxygen Species Production by Modulating Neutrophil-Platelet Interaction and CD11b Expression ▿ † ‡

    doi: 10.1128/IAI.00777-10

    Figure Lengend Snippet: LXA 4 does not affect P. gingivalis -induced ROS production in isolated neutrophils. Isolated polymorphonuclear cells (PMN), consisting predominantly of neutrophils, were preincubated for 15 min at 37°C in the presence or absence of LXA 4 (1 or 100

    Article Snippet: In short, whole blood was incubated at 37°C in the presence or absence of LXA4 (1, 100, 250, or 500 nM) for 15 min and subsequently stimulated with P. gingivalis (1 × 107 CFU/ml blood) for 10 min. To detect the total CD11b expression, an R-phycoerythrin (RPE)-conjugated anti-human CD11b antibody (mouse monoclonal antibody; clone 2LPM19c; Dako, Glostrup, Denmark) was added 5 min after the onset of bacterial stimulation.

    Techniques: Isolation

    LXA 4 does not affect P. gingivalis -induced aggregation of isolated platelets. Isolated platelets were preincubated for 15 min at 37°C in the presence or absence of LXA 4 (1 or 100 nM) and then monitored for aggregation upon stimulation with P.

    Journal: Infection and Immunity

    Article Title: Lipoxin A4 Inhibits Porphyromonas gingivalis-Induced Aggregation and Reactive Oxygen Species Production by Modulating Neutrophil-Platelet Interaction and CD11b Expression ▿-Induced Aggregation and Reactive Oxygen Species Production by Modulating Neutrophil-Platelet Interaction and CD11b Expression ▿ †-Induced Aggregation and Reactive Oxygen Species Production by Modulating Neutrophil-Platelet Interaction and CD11b Expression ▿ † ‡

    doi: 10.1128/IAI.00777-10

    Figure Lengend Snippet: LXA 4 does not affect P. gingivalis -induced aggregation of isolated platelets. Isolated platelets were preincubated for 15 min at 37°C in the presence or absence of LXA 4 (1 or 100 nM) and then monitored for aggregation upon stimulation with P.

    Article Snippet: In short, whole blood was incubated at 37°C in the presence or absence of LXA4 (1, 100, 250, or 500 nM) for 15 min and subsequently stimulated with P. gingivalis (1 × 107 CFU/ml blood) for 10 min. To detect the total CD11b expression, an R-phycoerythrin (RPE)-conjugated anti-human CD11b antibody (mouse monoclonal antibody; clone 2LPM19c; Dako, Glostrup, Denmark) was added 5 min after the onset of bacterial stimulation.

    Techniques: Isolation

    LXA 4 inhibits P. gingivalis -induced activation of Rac2 and Cdc42 in neutrophils. Neutrophils (5 × 10 6 /sample) were preincubated in the absence or presence of 5 nM LXA 4 for 5 min at 37°C and stimulated with P. gingivalis (6 × 10

    Journal: Infection and Immunity

    Article Title: Lipoxin A4 Inhibits Porphyromonas gingivalis-Induced Aggregation and Reactive Oxygen Species Production by Modulating Neutrophil-Platelet Interaction and CD11b Expression ▿-Induced Aggregation and Reactive Oxygen Species Production by Modulating Neutrophil-Platelet Interaction and CD11b Expression ▿ †-Induced Aggregation and Reactive Oxygen Species Production by Modulating Neutrophil-Platelet Interaction and CD11b Expression ▿ † ‡

    doi: 10.1128/IAI.00777-10

    Figure Lengend Snippet: LXA 4 inhibits P. gingivalis -induced activation of Rac2 and Cdc42 in neutrophils. Neutrophils (5 × 10 6 /sample) were preincubated in the absence or presence of 5 nM LXA 4 for 5 min at 37°C and stimulated with P. gingivalis (6 × 10

    Article Snippet: In short, whole blood was incubated at 37°C in the presence or absence of LXA4 (1, 100, 250, or 500 nM) for 15 min and subsequently stimulated with P. gingivalis (1 × 107 CFU/ml blood) for 10 min. To detect the total CD11b expression, an R-phycoerythrin (RPE)-conjugated anti-human CD11b antibody (mouse monoclonal antibody; clone 2LPM19c; Dako, Glostrup, Denmark) was added 5 min after the onset of bacterial stimulation.

    Techniques: Activation Assay

    Effect of the Nano-MIX on the multi-species biofilms of S. mutans , F. nucleatum , A. actinomycetemcomitans , and P. gingivalis at 24 h. The confocal scanning laser microscopy (CLSM) ( A , B ) and scanning electron microscopy (SEM) images ( C , D ) showing comparative antibacterial effects of the Nano-MIX treatment ( B , D ) on the mixed-species oral biofilms with reference to the blank nanoparticles ( A , C ), respectively.

    Journal: Nanomaterials

    Article Title: Synergistic Antibacterial Effects of Nanoparticles Encapsulated with Scutellaria baicalensis and Pure Chlorhexidine on Oral Bacterial Biofilms

    doi: 10.3390/nano6040061

    Figure Lengend Snippet: Effect of the Nano-MIX on the multi-species biofilms of S. mutans , F. nucleatum , A. actinomycetemcomitans , and P. gingivalis at 24 h. The confocal scanning laser microscopy (CLSM) ( A , B ) and scanning electron microscopy (SEM) images ( C , D ) showing comparative antibacterial effects of the Nano-MIX treatment ( B , D ) on the mixed-species oral biofilms with reference to the blank nanoparticles ( A , C ), respectively.

    Article Snippet: The multi-species biofilm samples of S. mutans , F. nucleatum , A. actinomycetemcomitans and P. gingivalis were subjected to further assessments by both SEM (Hitachi, Tokyo, Japan) and CLSM (Olympus, Tokyo, Japan) at 24 h as previously described [ ], in order to confirm the in vitro findngs.

    Techniques: Microscopy, Confocal Laser Scanning Microscopy, Electron Microscopy

    Immunohistochemical analysis of SDF-1α and CXCR4 in P. gingivalis challenged experimental rat periodontal inflammation. SDF-1α and CXCR4 were abundantly expressed in the P. gingivalis challenged rat tissues. Immunohistochemical analysis revealed a higher expression of SDF-1α and CXCR4 in P. gingivalis challenged rat gingiva (Fig. 4a; Scale bars, 20 μM) and PDL (Fig. 4b; Scale bars, 100 μM) compared to the control

    Journal: BMC Oral Health

    Article Title: Modulation of stromal cell-derived factor 1 alpha (SDF-1α) and its receptor CXCR4 in Porphyromonas gingivalis-induced periodontal inflammation

    doi: 10.1186/s12903-016-0250-8

    Figure Lengend Snippet: Immunohistochemical analysis of SDF-1α and CXCR4 in P. gingivalis challenged experimental rat periodontal inflammation. SDF-1α and CXCR4 were abundantly expressed in the P. gingivalis challenged rat tissues. Immunohistochemical analysis revealed a higher expression of SDF-1α and CXCR4 in P. gingivalis challenged rat gingiva (Fig. 4a; Scale bars, 20 μM) and PDL (Fig. 4b; Scale bars, 100 μM) compared to the control

    Article Snippet: Bacterial cells were grown under anaerobic conditions (85 % N2 , 10 % H2 and 5 % CO2 ) at 37 °C for 24 h. LPS from P. gingivalis ATCC 33277 was obtained from P. gingivalis according to the manufacturers’ instructions (iNtRON Biotechnology, Kyungki-Do, Korea).

    Techniques: Immunohistochemistry, Expressing

    a Effect of LPS on the expression of SDF-1α and CXCR4 in HGF-1 cells. HGF-1 cells were incubated with different concentrations of LPS from P. gingivalis for 24 h. The cell lysates were then assayed to determine the expression of SDF-1α and CXCR4, Akt and NF-kβ p65, and the phosphorylation of Akt and NF-kβ p65 using Western blots. Membranes were stripped and re-probed with an anti-β-actin antibody as a loading control. Protein bands were quantified by densitometric analyses. The results are expressed as means ± S.D. (* p

    Journal: BMC Oral Health

    Article Title: Modulation of stromal cell-derived factor 1 alpha (SDF-1α) and its receptor CXCR4 in Porphyromonas gingivalis-induced periodontal inflammation

    doi: 10.1186/s12903-016-0250-8

    Figure Lengend Snippet: a Effect of LPS on the expression of SDF-1α and CXCR4 in HGF-1 cells. HGF-1 cells were incubated with different concentrations of LPS from P. gingivalis for 24 h. The cell lysates were then assayed to determine the expression of SDF-1α and CXCR4, Akt and NF-kβ p65, and the phosphorylation of Akt and NF-kβ p65 using Western blots. Membranes were stripped and re-probed with an anti-β-actin antibody as a loading control. Protein bands were quantified by densitometric analyses. The results are expressed as means ± S.D. (* p

    Article Snippet: Bacterial cells were grown under anaerobic conditions (85 % N2 , 10 % H2 and 5 % CO2 ) at 37 °C for 24 h. LPS from P. gingivalis ATCC 33277 was obtained from P. gingivalis according to the manufacturers’ instructions (iNtRON Biotechnology, Kyungki-Do, Korea).

    Techniques: Expressing, Incubation, Western Blot

    Schematic illustration of extracellular oligopeptide metabolism in P. gingivalis . The metabolic pathway of P. gingivalis from the extracellular polypeptides, di- and tri-peptide incorporation, amino acid metabolism, and excretion as short-chain fatty acids, are schematically illustrated [10] , [15] . Amino acids, except for Ser and Thr, are mainly transported as di- and tri-peptides via oligopeptide transporters [10] . Rgp and Kgp are mainly localized on the outer membrane (OM), while DPPs, PtpA, and AOP are located in periplasmic space [23] , [24] , [25] . Scissors indicate peptidases, which cleave peptide bonds at specific positions. Stars represent acylaminoacyl groups at the N-terminus. IM, inner membrane.

    Journal: The Japanese Dental Science Review

    Article Title: Exopeptidases and gingipains in Porphyromonas gingivalis as prerequisites for its amino acid metabolism

    doi: 10.1016/j.jdsr.2015.08.002

    Figure Lengend Snippet: Schematic illustration of extracellular oligopeptide metabolism in P. gingivalis . The metabolic pathway of P. gingivalis from the extracellular polypeptides, di- and tri-peptide incorporation, amino acid metabolism, and excretion as short-chain fatty acids, are schematically illustrated [10] , [15] . Amino acids, except for Ser and Thr, are mainly transported as di- and tri-peptides via oligopeptide transporters [10] . Rgp and Kgp are mainly localized on the outer membrane (OM), while DPPs, PtpA, and AOP are located in periplasmic space [23] , [24] , [25] . Scissors indicate peptidases, which cleave peptide bonds at specific positions. Stars represent acylaminoacyl groups at the N-terminus. IM, inner membrane.

    Article Snippet: Additional note To activate studies of DPPs of P. gingivalis and other oral bacteria, we contacted the Peptide Institute (Osaka, Japan) to produce DPP substrates.

    Techniques:

    P. gingivalis bypasses the epithelial barrier to reach FBs located at the core of the MT leading to MT destruction. SEM imaging of ( A , B ) P. gingivalis culture, ( C ) P. gingivalis infection and intracellular invasion of the MT surface, ( D ) P. gingivalis proliferating on MT surface and EC exfoliation, ( E ) uninfected MT, ( F ) MT stimulated with LPS, ( G ) MT infected with P. gingivalis for 6 h and ( H ) for 24 h. Green arrows indicate P. gingivalis infection in MT and/or cell invasion; blue arrows indicate the disruption of epithelial barrier, multiple apoptotic cells and an acute cellular injury in all strata of the MT – EC and FB-; white arrow indicates proliferating EC at the surface of MT.

    Journal: Scientific Reports

    Article Title: Porphyromonas gingivalis bypasses epithelial barrier and modulates fibroblastic inflammatory response in an in vitro 3D spheroid model

    doi: 10.1038/s41598-018-33267-4

    Figure Lengend Snippet: P. gingivalis bypasses the epithelial barrier to reach FBs located at the core of the MT leading to MT destruction. SEM imaging of ( A , B ) P. gingivalis culture, ( C ) P. gingivalis infection and intracellular invasion of the MT surface, ( D ) P. gingivalis proliferating on MT surface and EC exfoliation, ( E ) uninfected MT, ( F ) MT stimulated with LPS, ( G ) MT infected with P. gingivalis for 6 h and ( H ) for 24 h. Green arrows indicate P. gingivalis infection in MT and/or cell invasion; blue arrows indicate the disruption of epithelial barrier, multiple apoptotic cells and an acute cellular injury in all strata of the MT – EC and FB-; white arrow indicates proliferating EC at the surface of MT.

    Article Snippet: At the day of the experiment, cells were washed twice with PBS and were either infected for 24 h with P. gingivalis at a multiplicity of infection (MOI) of 100 or stimulated with ultrapure P. gingivalis LPS (1 μg/ml) (InvivoGen, San Diego, CA, USA).

    Techniques: Imaging, Infection

    Effect of P. gingivalis 381 LPS on HASMC TFPI production

    Journal: Thrombosis research

    Article Title: Porphyromonas gingivalis infection and prothrombotic effects in human aortic smooth muscle cells

    doi: 10.1016/j.thromres.2008.07.008

    Figure Lengend Snippet: Effect of P. gingivalis 381 LPS on HASMC TFPI production

    Article Snippet: In some experiments, heat-killed P. gingivalis 381 and ultrapure P. gingivalis lipopolysacharide (LPS) (Invivogen, San Diego, CA) were used as controls.

    Techniques:

    EMV Pg induce pro-inflammatory and pro-oxidative protein expression similar to P. gingivalis infection. ( A ) TNF-α secretion in supernatant by EC in response to P. gingivalis ( Pg ) (MOI:100) and EMV Pg (5, 20 and 30 nM) for 24 h was measured by ELISA. ( B ) Intra-cellular protein expression of eNOS, P21, ICAM-1, CDK4, P53, VCAM, iNOS and SOD-1 in response to P. gingivalis ( Pg ) (MOI:100) and EMV Pg (5, 20 and 30 nM) for 24 h was evaluated by Western Blot. All data were expressed as mean ± SD from 3 independent experiments and normalized against internal control β-actin. * p

    Journal: Scientific Reports

    Article Title: Porphyromonas gingivalis triggers the shedding of inflammatory endothelial microvesicles that act as autocrine effectors of endothelial dysfunction

    doi: 10.1038/s41598-020-58374-z

    Figure Lengend Snippet: EMV Pg induce pro-inflammatory and pro-oxidative protein expression similar to P. gingivalis infection. ( A ) TNF-α secretion in supernatant by EC in response to P. gingivalis ( Pg ) (MOI:100) and EMV Pg (5, 20 and 30 nM) for 24 h was measured by ELISA. ( B ) Intra-cellular protein expression of eNOS, P21, ICAM-1, CDK4, P53, VCAM, iNOS and SOD-1 in response to P. gingivalis ( Pg ) (MOI:100) and EMV Pg (5, 20 and 30 nM) for 24 h was evaluated by Western Blot. All data were expressed as mean ± SD from 3 independent experiments and normalized against internal control β-actin. * p

    Article Snippet: For comparative purposes, in some experiments, EC were stimulated with P. gingivalis ultrapure lipopolysaccharide (Pg- LPS) (1 μg/ml) (InvivoGen, San Diego, CA, USA) for 24 h.

    Techniques: Expressing, Infection, Enzyme-linked Immunosorbent Assay, Western Blot

    Exposure to EMV Pg modulates significantly inflammatory pathways related to kinases activation. Analysis of kinases activation induced by P. gingivalis infection ( Pg ) (MOI:100) and EMV Pg (30 nM) for 24 h evaluated by phospho-kinase array. The density of spots was measured by MyImage TM Analysis Softwate 2.0 (Thermofisher) for each molecule and each condition. ( A ) Graphical representation of the kinases expression in untreated EC, in EC following P. gingivalis infection ( Pg ) (MOI:100) and EMV Pg (30 nM) stimulation for 24 h. ( B ) Heat-map representation of the kinases expression normalized against untreated control (untreated EC).

    Journal: Scientific Reports

    Article Title: Porphyromonas gingivalis triggers the shedding of inflammatory endothelial microvesicles that act as autocrine effectors of endothelial dysfunction

    doi: 10.1038/s41598-020-58374-z

    Figure Lengend Snippet: Exposure to EMV Pg modulates significantly inflammatory pathways related to kinases activation. Analysis of kinases activation induced by P. gingivalis infection ( Pg ) (MOI:100) and EMV Pg (30 nM) for 24 h evaluated by phospho-kinase array. The density of spots was measured by MyImage TM Analysis Softwate 2.0 (Thermofisher) for each molecule and each condition. ( A ) Graphical representation of the kinases expression in untreated EC, in EC following P. gingivalis infection ( Pg ) (MOI:100) and EMV Pg (30 nM) stimulation for 24 h. ( B ) Heat-map representation of the kinases expression normalized against untreated control (untreated EC).

    Article Snippet: For comparative purposes, in some experiments, EC were stimulated with P. gingivalis ultrapure lipopolysaccharide (Pg- LPS) (1 μg/ml) (InvivoGen, San Diego, CA, USA) for 24 h.

    Techniques: Activation Assay, Infection, Expressing

    P. gingivalis promotes EMV shedding and alters endothelial cell viability. ( A ) The generation of EMV from naïve EC (control) and following 24 h of infection with P. gingivalis ( Pg) (MOI = 100) or stimulation with Pg -LPS (1μg/ml) was measured in the supernatant by prothrombinase assay. Concentrations are represented by mean +/− SD from 3 independent experiments; * p

    Journal: Scientific Reports

    Article Title: Porphyromonas gingivalis triggers the shedding of inflammatory endothelial microvesicles that act as autocrine effectors of endothelial dysfunction

    doi: 10.1038/s41598-020-58374-z

    Figure Lengend Snippet: P. gingivalis promotes EMV shedding and alters endothelial cell viability. ( A ) The generation of EMV from naïve EC (control) and following 24 h of infection with P. gingivalis ( Pg) (MOI = 100) or stimulation with Pg -LPS (1μg/ml) was measured in the supernatant by prothrombinase assay. Concentrations are represented by mean +/− SD from 3 independent experiments; * p

    Article Snippet: For comparative purposes, in some experiments, EC were stimulated with P. gingivalis ultrapure lipopolysaccharide (Pg- LPS) (1 μg/ml) (InvivoGen, San Diego, CA, USA) for 24 h.

    Techniques: Infection

    EMV Pg induce expression of atherothrombosis and oxidative stress markers expression. ( A ) The mRNA expression of endothelial markers VCAM, ICAM and Tissular factor (TF) in EC exposed to P. gingivalis ( Pg ) (MOI:100) and EMV Pg (5, 10, 20 and 30 nM) for 24 h was measured by qRT-PCR. ( B ) The gene expression of oxidative stress markers eNOS, iNOS and SOD-1 in EC exposed to P. gingivalis ( Pg ) (MOI:100) and EMV Pg (5, 10, 20 and 30 nM) for 24 h. All data were expressed as mean ± SD from 3 independent experiments. * p

    Journal: Scientific Reports

    Article Title: Porphyromonas gingivalis triggers the shedding of inflammatory endothelial microvesicles that act as autocrine effectors of endothelial dysfunction

    doi: 10.1038/s41598-020-58374-z

    Figure Lengend Snippet: EMV Pg induce expression of atherothrombosis and oxidative stress markers expression. ( A ) The mRNA expression of endothelial markers VCAM, ICAM and Tissular factor (TF) in EC exposed to P. gingivalis ( Pg ) (MOI:100) and EMV Pg (5, 10, 20 and 30 nM) for 24 h was measured by qRT-PCR. ( B ) The gene expression of oxidative stress markers eNOS, iNOS and SOD-1 in EC exposed to P. gingivalis ( Pg ) (MOI:100) and EMV Pg (5, 10, 20 and 30 nM) for 24 h. All data were expressed as mean ± SD from 3 independent experiments. * p

    Article Snippet: For comparative purposes, in some experiments, EC were stimulated with P. gingivalis ultrapure lipopolysaccharide (Pg- LPS) (1 μg/ml) (InvivoGen, San Diego, CA, USA) for 24 h.

    Techniques: Expressing, Quantitative RT-PCR

    EMV Pg trigger inflammatory endothelial response. ( A ) The mRNA expression of inflammatory markers TNF-α, IL-6 and IL-8 in EC exposed to P. gingivalis ( Pg ) (MOI:100) and EMV Pg (5, 10, 20 and 30 nM) for 24 h was measured by qRT-PCR. ( B ) The gene expression of cell cyle related markers p53, p21 and CDK4 EC in EC exposed to P. gingivalis ( Pg ) (MOI:100) and EMV Pg (5, 10, 20 and 30 nM) for 24 h. All data were expressed as mean ± SD from 3 independent experiments. * p

    Journal: Scientific Reports

    Article Title: Porphyromonas gingivalis triggers the shedding of inflammatory endothelial microvesicles that act as autocrine effectors of endothelial dysfunction

    doi: 10.1038/s41598-020-58374-z

    Figure Lengend Snippet: EMV Pg trigger inflammatory endothelial response. ( A ) The mRNA expression of inflammatory markers TNF-α, IL-6 and IL-8 in EC exposed to P. gingivalis ( Pg ) (MOI:100) and EMV Pg (5, 10, 20 and 30 nM) for 24 h was measured by qRT-PCR. ( B ) The gene expression of cell cyle related markers p53, p21 and CDK4 EC in EC exposed to P. gingivalis ( Pg ) (MOI:100) and EMV Pg (5, 10, 20 and 30 nM) for 24 h. All data were expressed as mean ± SD from 3 independent experiments. * p

    Article Snippet: For comparative purposes, in some experiments, EC were stimulated with P. gingivalis ultrapure lipopolysaccharide (Pg- LPS) (1 μg/ml) (InvivoGen, San Diego, CA, USA) for 24 h.

    Techniques: Expressing, Quantitative RT-PCR

    (A) IL-6 and TNFα levels in tissue culture supernatants from unstimulated (no LPS) and P. gingivalis LPS-stimulated cell lines, and (B) unstimulated (no LPS) and E. coli LPS-stimulated cell lines. (C) Anti-TLR4 antibody suppressed the secretion of IL-6 when stimulated by E. coli but not P. gingivalis LPS, implying different signaling mechanism mediated by LPS from those organisms. Data are mean values ± SEM of 2-3 experiments. (D) P. gingivalis LPS stimulation of three separate gingival cell lines, as indicated, resulted in a 2.3 – 8.4 fold increase in CCL25 RNA levels compared to non-stimulated (PBS) cells. Gene expression levels were determined for each sample relative to the 18s gene expression of that sample as described in the Materials and methods. Since all P. gingivalis LPS stimulated cultures had greater CCL25 gene expression than non-stimulated cultures, gene expression levels for non-stimulated cultures were assigned a value of 1.0 and gene expression levels of stimulated cultures were expressed as fold increase over non-stimulated cultures.

    Journal: Journal of periodontal research

    Article Title: Porphyromonas gingivalis lipopolysaccharide induces tumor necrosis factor-α and interleukin-6 (IL-6) secretion and CCL25 gene expression in mouse primary gingival cell lines: IL-6-driven activation of CCL2

    doi:

    Figure Lengend Snippet: (A) IL-6 and TNFα levels in tissue culture supernatants from unstimulated (no LPS) and P. gingivalis LPS-stimulated cell lines, and (B) unstimulated (no LPS) and E. coli LPS-stimulated cell lines. (C) Anti-TLR4 antibody suppressed the secretion of IL-6 when stimulated by E. coli but not P. gingivalis LPS, implying different signaling mechanism mediated by LPS from those organisms. Data are mean values ± SEM of 2-3 experiments. (D) P. gingivalis LPS stimulation of three separate gingival cell lines, as indicated, resulted in a 2.3 – 8.4 fold increase in CCL25 RNA levels compared to non-stimulated (PBS) cells. Gene expression levels were determined for each sample relative to the 18s gene expression of that sample as described in the Materials and methods. Since all P. gingivalis LPS stimulated cultures had greater CCL25 gene expression than non-stimulated cultures, gene expression levels for non-stimulated cultures were assigned a value of 1.0 and gene expression levels of stimulated cultures were expressed as fold increase over non-stimulated cultures.

    Article Snippet: Reagents used in this study included ultrapure P. gingivalis LPS (Invitrogen; San Diego; CA) and E. coli LPS (Sigma); recombinant IL-6 (rIL-6) ( < 0.01 ng endotoxin per μg cytokine (eBioscience; San Diego, CA); sIL-6R ( < 0.0 EU endotoxin per 1 μg cytokine receptor) (R & D Systems; Minneapolis, MN); two preparations of sgp130: recombinant human sgp130 ( < 1.0 EU endotoxin per 1 μg receptor) (R & D Systems), and recombinant mouse sgp130/Fc chimera ( < 1.0 ng endotoxin per 1 μg receptor) (R & D systems).

    Techniques: Expressing

    Validation of the germfree-rat model of P. gingivalis -induced alveolar bone loss in rats dually infected with P. gingivalis 381 (Pg381) and the S. gordonii carrier strain (Sg Carrier, Pg251). The mean alveolar bone loss (CEJ:ABC) was calculated as the mean difference between the CEJ and the ABC in millimeters per site for each group ± the standard error of the mean ( n = 8). Groups infected with P. gingivalis 381 alone (Pg381) or P. gingivalis and the S. gordonii carrier (Sg Carrier/Pg381) showed significant alveolar bone loss compared to groups colonized with the S. gordonii carrier alone (Sg Carrier) or sham infected (***, P

    Journal: Infection and Immunity

    Article Title: Oral Immunization with Recombinant Streptococcus gordonii Expressing Porphyromonas gingivalis FimA Domains

    doi: 10.1128/IAI.69.5.2928-2934.2001

    Figure Lengend Snippet: Validation of the germfree-rat model of P. gingivalis -induced alveolar bone loss in rats dually infected with P. gingivalis 381 (Pg381) and the S. gordonii carrier strain (Sg Carrier, Pg251). The mean alveolar bone loss (CEJ:ABC) was calculated as the mean difference between the CEJ and the ABC in millimeters per site for each group ± the standard error of the mean ( n = 8). Groups infected with P. gingivalis 381 alone (Pg381) or P. gingivalis and the S. gordonii carrier (Sg Carrier/Pg381) showed significant alveolar bone loss compared to groups colonized with the S. gordonii carrier alone (Sg Carrier) or sham infected (***, P

    Article Snippet: Briefly, P. gingivalis 381 was grown in half-strength brain heart infusion (18 mg/ml; Difco) supplemented with 5 mg of yeast extract per ml, 5 μg of hemin per ml, and 0.2 μg of menadione per ml and buffered at pH 7.4 under anaerobic conditions (anaerobic chamber; Forma Scientific, Marietta, Ohio) for 48 h. Cells were harvested after 2 days and washed with PBS, and a bacterial suspension for inoculation was made in 5% carboxymethyl cellulose.,

    Techniques: Infection

    PLNC8 αβ binds to P. gingivalis in a dose-dependent manner. Binding of P. gingivalis to PLNC8 αβ and to anti- P.gingivalis antibodies was analyzed by SPR. Both P. gingivalis ATCC 33277 ( a ) and W50 ( b ) were found to bind to immobilized PLNC8 αβ (280 nM). The binding was verified by pre-incubating the bacteria with different concentrations of soluble PLNC8 αβ prior to analysis, which resulted in a significantly reduced binding to the immobilized bacteriocins. Pre-incubation of P. gingivalis ATCC 33277 ( c ) and W50 ( d ) with increasing concentrations of soluble PLNC8 αβ prior to analysis reduced the bacterial binding to anti- P. gingivalis antibodies in a dose-dependent manner. Results are presented from three independent experiments. * p

    Journal: BMC Microbiology

    Article Title: Antibacterial effects of Lactobacillus and bacteriocin PLNC8 αβ on the periodontal pathogen Porphyromonas gingivalis

    doi: 10.1186/s12866-016-0810-8

    Figure Lengend Snippet: PLNC8 αβ binds to P. gingivalis in a dose-dependent manner. Binding of P. gingivalis to PLNC8 αβ and to anti- P.gingivalis antibodies was analyzed by SPR. Both P. gingivalis ATCC 33277 ( a ) and W50 ( b ) were found to bind to immobilized PLNC8 αβ (280 nM). The binding was verified by pre-incubating the bacteria with different concentrations of soluble PLNC8 αβ prior to analysis, which resulted in a significantly reduced binding to the immobilized bacteriocins. Pre-incubation of P. gingivalis ATCC 33277 ( c ) and W50 ( d ) with increasing concentrations of soluble PLNC8 αβ prior to analysis reduced the bacterial binding to anti- P. gingivalis antibodies in a dose-dependent manner. Results are presented from three independent experiments. * p

    Article Snippet: The lipid composition (5:95 POPS: POPC) was chosen to mimic the zeta potential of microvesicles present in the supernatant retrieved cultures of P. gingivalis W50 (-24.8 mV) (Fig. ).

    Techniques: Binding Assay, SPR Assay, Incubation

    Bacteriocin PLNC8 αβ from L. plantarum NC8 is efficient against P. gingivalis . The antimicrobial activity of PLNC8 αβ on wild type (WT) P. gingivalis ATCC 33277 ( a ) and W50 ( b ), respectively, was visualized using the fluorescent dye Sytox® Green. Images were acquired using Olympus BX41 at 40× magnification. The antimicrobial effect of PLNC8 αβ was rapid and a significant number of P. gingivalis cells could fluoresce already after 1 min, indicating damaged membranes. Representative images and quantitative data of at least three independent experiments are shown. Quantitative data were normalized and the controls at each time point were set to 1. *** p

    Journal: BMC Microbiology

    Article Title: Antibacterial effects of Lactobacillus and bacteriocin PLNC8 αβ on the periodontal pathogen Porphyromonas gingivalis

    doi: 10.1186/s12866-016-0810-8

    Figure Lengend Snippet: Bacteriocin PLNC8 αβ from L. plantarum NC8 is efficient against P. gingivalis . The antimicrobial activity of PLNC8 αβ on wild type (WT) P. gingivalis ATCC 33277 ( a ) and W50 ( b ), respectively, was visualized using the fluorescent dye Sytox® Green. Images were acquired using Olympus BX41 at 40× magnification. The antimicrobial effect of PLNC8 αβ was rapid and a significant number of P. gingivalis cells could fluoresce already after 1 min, indicating damaged membranes. Representative images and quantitative data of at least three independent experiments are shown. Quantitative data were normalized and the controls at each time point were set to 1. *** p

    Article Snippet: The lipid composition (5:95 POPS: POPC) was chosen to mimic the zeta potential of microvesicles present in the supernatant retrieved cultures of P. gingivalis W50 (-24.8 mV) (Fig. ).

    Techniques: Activity Assay

    Zeta potential and size of liposomes and W50 microvesicles. The zeta potential and size of liposomes with different lipid composition was measured to identify the best match with microvesicles from P. gingivalis W50. All measurements are done in triplicates

    Journal: BMC Microbiology

    Article Title: Antibacterial effects of Lactobacillus and bacteriocin PLNC8 αβ on the periodontal pathogen Porphyromonas gingivalis

    doi: 10.1186/s12866-016-0810-8

    Figure Lengend Snippet: Zeta potential and size of liposomes and W50 microvesicles. The zeta potential and size of liposomes with different lipid composition was measured to identify the best match with microvesicles from P. gingivalis W50. All measurements are done in triplicates

    Article Snippet: The lipid composition (5:95 POPS: POPC) was chosen to mimic the zeta potential of microvesicles present in the supernatant retrieved cultures of P. gingivalis W50 (-24.8 mV) (Fig. ).

    Techniques:

    Biofilm formation by homotypic P. gingivalis 33277, ECF sigma factor mutant and complemented mutant strain.  The strains were grown on enriched BHI broth anaerobically at 37°C in collagen type-I-coated 12-well flat-bottom microplates. After 48 h of cultivation, the organized biofilm mass was evaluated by staining with crystal violet.  (a)  Biofilm formation of 33277, PGN_0274 mutant and the complemented mutant strain were compared.  (b)  Biofilm formation of 33277, PGN_1740 mutant and the complemented mutant strain were compared. Biofilm formation determined by crystal violet staining and adjusted for growth (A 600  units per OD 660  unit). The data shown are mean ± SD of triplicate experiments. *,  p

    Journal: BMC Oral Health

    Article Title: Role of extracytoplasmic function sigma factors in biofilm formation of Porphyromonas gingivalis

    doi: 10.1186/1472-6831-15-4

    Figure Lengend Snippet: Biofilm formation by homotypic P. gingivalis 33277, ECF sigma factor mutant and complemented mutant strain. The strains were grown on enriched BHI broth anaerobically at 37°C in collagen type-I-coated 12-well flat-bottom microplates. After 48 h of cultivation, the organized biofilm mass was evaluated by staining with crystal violet. (a) Biofilm formation of 33277, PGN_0274 mutant and the complemented mutant strain were compared. (b) Biofilm formation of 33277, PGN_1740 mutant and the complemented mutant strain were compared. Biofilm formation determined by crystal violet staining and adjusted for growth (A 600 units per OD 660 unit). The data shown are mean ± SD of triplicate experiments. *, p

    Article Snippet: Additional file 1: Comparisons of biofilm treated with ethanol or SDS. (PPTX 456 KB) Additional file 2: Biofilm formation by homotypic P. gingivalis 33277 or ECF sigma factor mutants using non-coated microplate. (PPTX 80 KB) Additional file 3: The RNA expression of fimS in P. gingivalis 33277, PGN _ 1740 mutant and complemented mutant strain. (PPTX 101 KB) Additional file 4: Protein profile on an SDS-PAGE gel. (PPTX 343 KB)

    Techniques: Mutagenesis, Staining

    Growth of P. gingivalis 33277 and ECF sigma factor mutants. Growth curves of P. gingivalis 33277 (wild-type; circle), PGN_0274 mutant (KDP314; open rectangle), PGN_0319 mutant (KDP315; closed rectangle), PGN_0450 mutant (KDP316; diamond), PGN_0970 mutant (KDP317; triangle), PGN_1740 mutant (KDP319; cross) in enriched BHI broth. The data shown are mean ± SD of triplicate experiments.

    Journal: BMC Oral Health

    Article Title: Role of extracytoplasmic function sigma factors in biofilm formation of Porphyromonas gingivalis

    doi: 10.1186/1472-6831-15-4

    Figure Lengend Snippet: Growth of P. gingivalis 33277 and ECF sigma factor mutants. Growth curves of P. gingivalis 33277 (wild-type; circle), PGN_0274 mutant (KDP314; open rectangle), PGN_0319 mutant (KDP315; closed rectangle), PGN_0450 mutant (KDP316; diamond), PGN_0970 mutant (KDP317; triangle), PGN_1740 mutant (KDP319; cross) in enriched BHI broth. The data shown are mean ± SD of triplicate experiments.

    Article Snippet: Additional file 1: Comparisons of biofilm treated with ethanol or SDS. (PPTX 456 KB) Additional file 2: Biofilm formation by homotypic P. gingivalis 33277 or ECF sigma factor mutants using non-coated microplate. (PPTX 80 KB) Additional file 3: The RNA expression of fimS in P. gingivalis 33277, PGN _ 1740 mutant and complemented mutant strain. (PPTX 101 KB) Additional file 4: Protein profile on an SDS-PAGE gel. (PPTX 343 KB)

    Techniques: Mutagenesis

    Biofilm formation by homotypic P. gingivalis 33277 or ECF sigma factor mutants. The strains were grown on enriched BHI broth anaerobically at 37°C in collagen type-I-coated 12-well flat-bottom microplates. After 48 h of cultivation, the organized biofilm mass was evaluated by staining with crystal violet. (a) The photographs are a representative sample of each experimental strain. (b) Biofilm formation determined by crystal violet staining and adjusted for growth (A 600 units per OD 660 unit). The data shown are mean ± SD of triplicate experiments. ***, p

    Journal: BMC Oral Health

    Article Title: Role of extracytoplasmic function sigma factors in biofilm formation of Porphyromonas gingivalis

    doi: 10.1186/1472-6831-15-4

    Figure Lengend Snippet: Biofilm formation by homotypic P. gingivalis 33277 or ECF sigma factor mutants. The strains were grown on enriched BHI broth anaerobically at 37°C in collagen type-I-coated 12-well flat-bottom microplates. After 48 h of cultivation, the organized biofilm mass was evaluated by staining with crystal violet. (a) The photographs are a representative sample of each experimental strain. (b) Biofilm formation determined by crystal violet staining and adjusted for growth (A 600 units per OD 660 unit). The data shown are mean ± SD of triplicate experiments. ***, p

    Article Snippet: Additional file 1: Comparisons of biofilm treated with ethanol or SDS. (PPTX 456 KB) Additional file 2: Biofilm formation by homotypic P. gingivalis 33277 or ECF sigma factor mutants using non-coated microplate. (PPTX 80 KB) Additional file 3: The RNA expression of fimS in P. gingivalis 33277, PGN _ 1740 mutant and complemented mutant strain. (PPTX 101 KB) Additional file 4: Protein profile on an SDS-PAGE gel. (PPTX 343 KB)

    Techniques: Staining