p. gingivalis Search Results


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  • 99
    ATCC p gingivalis atcc 33277
    Pangenome overview of ATCC 33277, TDC60, and W83 strains, focusing on accessory and unique genomes. The central triangle represents the core genome, which has at least 1522 genes (see text for details). Each corner is a Porphyromonas <t>gingivalis</t> ( P. g. ) strain, with a pie chart showing the unique genome’s distribution of functions, with total and absolute counts shown. On each triangle side, stacked histograms show the accessory genome of the strains in the adjacent vertices. Total and absolute counts are shown, and the differences between strain numbers are due to paralogy
    P Gingivalis Atcc 33277, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 2730 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    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: 92/100, based on 347 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    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: 91/100, based on 192 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Illumina Inc p gingivalis
    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, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 92/100, based on 211 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    The Jackson Laboratory p gingivalis
    The frequency of total CD4 + T cells producing IL-17A was significantly higher in the CLN of C57BL/6J dams than in C57BL/6NCrl dams after oral inoculation with P. <t>gingivalis</t> . CD4 + T cells from MLN (A and B) or PaLN-CLN (C and D) of pregnant dams were stimulated with PMA-ionomycin and analyzed by flow cytometry for total expression of IL-17A or IFN-γ after P. gingivalis oral inoculation. The percentages of total CD4 + T cells producing cytokines across groups were compared using a Mann-Whitney nonparametric test, since the cytokine data did not have a normal distribution. Medians plus interquartile ranges are shown in the plot. C57BL/6J (B6/J) dams are represented by square symbols; C57BL/6NCrl (B6/NCrl, SFB + ) dams are represented by circular symbols.
    P Gingivalis, supplied by The Jackson Laboratory, used in various techniques. Bioz Stars score: 96/100, based on 56 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Becton Dickinson p gingivalis
    Scanning electron microscopy images of Porphyromonas <t>gingivalis</t> cells attached to the sandblasted large-grit acid-etched surface: (A, F) control group; (B, G) photodynamic therapy group; (C, H) brush alone group; (D, I) brushing with a LED light group; (E, J) brushing with a LED light and erythrosine group. LED: light-emitting diode.
    P Gingivalis, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 137 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    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: 92/100, based on 34 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    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: 93/100, based on 64 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    ATCC p gingivalis atcc 53978
    Distribution of PAD in various strains of P. <t>gingivalis</t> . The production of citrulline from BAEE was determined in cellular (open bar), supernatant (solid bar), and vesicle (hatched bar) fractions from HG66, <t>ATCC</t> 53978 (W50), and ATCC 33277 strains of P. gingivalis .
    P Gingivalis Atcc 53978, supplied by ATCC, used in various techniques. Bioz Stars score: 95/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    ATCC p gingivalis w83
    Genomic representation of the P. <t>gingivalis</t> proteome, showing changes in relative abundance for the P. gingivalis - F. nucleatum - S. gordonii / P. gingivalis comparison by spectral counting . Each dot represents a PGN ORF number in the order followed by the ATCC 33277 strain annotation. Color codes: red, over-expression in the P. gingivalis - F. nucleatum - S. gordonii community relative to P. gingivalis alone; green, under-expression in the community relative to P. gingivalis alone; yellow, protein was detected qualitatively, but did not change in abundance; gray, proteins that were qualitative non-detects; gaps indicate ORFs that were not common to both the ATCC 33277 and <t>W83</t> annotations according to a master cross-reference compiled by LANL (G. Xie, personal communication).
    P Gingivalis W83, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 147 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    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: 89/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    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: 92/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Promega p gingivalis
    Quantitative and qualitative biofilm assays. (A) Quantitative analysis of biofilm accretion by P. <t>gingivalis</t> strains and chimeras. Chimeric strains 2, 4, and 9 (dark bars) were compared to strain 33277TcEm for statistical analysis by t test. All other chimeras (light bars) were compared to W83TcEm. Results represent three independent experiments, each done in triplicate ( n = 9). Symbols * and # represent P values of
    P Gingivalis, supplied by Promega, used in various techniques. Bioz Stars score: 92/100, based on 116 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    InvivoGen ultrapure p gingivalis lps
    Effect of 25(OH)D 3 and 1,25(OH) 2 D 3 on the production of pro-inflammatory mediators by hPdLF with silenced VDR. Cells were transfected with either VDR siRNA or control siRNA and stimulated with P. <t>gingivalis</t> <t>LPS</t> ( Pg LPS, 1 µg/ml) or heat-killed P. gingivalis (hk Pg , 10 8 cells/ml) for 24 h in the presence or in the absence of 25(OH)D 3 or 1,25(OH) 2 D 3 . # means significantly different from control group (2 −▵▵Ct = 1). * means significantly different from cells stimulated with either P. gingivalis LPS or heat-killed P. gingivalis only
    Ultrapure P Gingivalis Lps, supplied by InvivoGen, used in various techniques. Bioz Stars score: 88/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Pangenome overview of ATCC 33277, TDC60, and W83 strains, focusing on accessory and unique genomes. The central triangle represents the core genome, which has at least 1522 genes (see text for details). Each corner is a Porphyromonas gingivalis ( P. g. ) strain, with a pie chart showing the unique genome’s distribution of functions, with total and absolute counts shown. On each triangle side, stacked histograms show the accessory genome of the strains in the adjacent vertices. Total and absolute counts are shown, and the differences between strain numbers are due to paralogy

    Journal: BMC Genomics

    Article Title: Genomic repeats, misassembly and reannotation: a case study with long-read resequencing of Porphyromonas gingivalis reference strains

    doi: 10.1186/s12864-017-4429-4

    Figure Lengend Snippet: Pangenome overview of ATCC 33277, TDC60, and W83 strains, focusing on accessory and unique genomes. The central triangle represents the core genome, which has at least 1522 genes (see text for details). Each corner is a Porphyromonas gingivalis ( P. g. ) strain, with a pie chart showing the unique genome’s distribution of functions, with total and absolute counts shown. On each triangle side, stacked histograms show the accessory genome of the strains in the adjacent vertices. Total and absolute counts are shown, and the differences between strain numbers are due to paralogy

    Article Snippet: The first band is 9 Kb, and the second is 4 Kb. b. Schematic representation (not to scale) of both copies (“a” and “b”) of CTnPg1 from P. gingivalis ATCC 33277.

    Techniques:

    Specific groups of metabolites influenced respiratory activity of each bacterial community. Principal Coordinate Analysis assisted to explain the grouping trends in the cluster analysis, and highlighted similarities in the respiratory activity among the selected oral bacteria at 24 h (A) and 48 h (B) . Aa, Aggregatibacter actinomycetemcomitans (ATCC 43718); Fn, Fusobacterium nucleatum (ATCC 10953); Pg, Porphyromonas gingivalis (ATCC 33277); Pi, Prevotella intermedia (ATCC 25611); Sm, Streptococcus mutans (ATCC 25175); Ssob, Streptococcus sobrinus (ATCC 33478); Tf, Tannerella forsythia (ATCC 43037); An, Actinomyces naeslundii (ATCC 51655); Csput, Capnocytophaga sputigena (ATCC 33612); Sgord, Streptococcus gordonii (ATCC 49818); Avisc, Actinomyces viscosus (ATCC 15987); Smitis, Streptococcus mitis (ATCC 49456); Vparv, Veillonella parvula (DSM 2007); Ssang, Streptococcus sanguinis (LMG 14657), and Ssal, Streptococcus salivarius strain TOVE-R.

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: In vitro Increased Respiratory Activity of Selected Oral Bacteria May Explain Competitive and Collaborative Interactions in the Oral Microbiome

    doi: 10.3389/fcimb.2017.00235

    Figure Lengend Snippet: Specific groups of metabolites influenced respiratory activity of each bacterial community. Principal Coordinate Analysis assisted to explain the grouping trends in the cluster analysis, and highlighted similarities in the respiratory activity among the selected oral bacteria at 24 h (A) and 48 h (B) . Aa, Aggregatibacter actinomycetemcomitans (ATCC 43718); Fn, Fusobacterium nucleatum (ATCC 10953); Pg, Porphyromonas gingivalis (ATCC 33277); Pi, Prevotella intermedia (ATCC 25611); Sm, Streptococcus mutans (ATCC 25175); Ssob, Streptococcus sobrinus (ATCC 33478); Tf, Tannerella forsythia (ATCC 43037); An, Actinomyces naeslundii (ATCC 51655); Csput, Capnocytophaga sputigena (ATCC 33612); Sgord, Streptococcus gordonii (ATCC 49818); Avisc, Actinomyces viscosus (ATCC 15987); Smitis, Streptococcus mitis (ATCC 49456); Vparv, Veillonella parvula (DSM 2007); Ssang, Streptococcus sanguinis (LMG 14657), and Ssal, Streptococcus salivarius strain TOVE-R.

    Article Snippet: Bacterial cultures and collection The following strains commonly present in the oral microbiome (Aas et al., ; Maddi and Scannapieco, ; Loozen et al., ) were obtained from the American Type Culture Collection (ATCC): A . actinomycetemcomitans (ATCC 43718), F. nucleatum (ATCC 10953), P. gingivalis (ATCC 33277), P. intermedia (ATCC 25611), S. mutans (ATCC 25175), S. sobrinus (ATCC 33478), T. forsythia (ATCC 43037), Actinomyces naeslundii (ATCC 51655), Capnocytophaga sputigena (ATCC 33612), Streptococcus gordonii (ATCC 49818), Actinomyces viscosus (ATCC 15987), and S. mitis (ATCC 49456).

    Techniques: Activity Assay

    Binding of P. gingivalis ATCC 33277 to human RBCs. (A) Flow cytometry histogram showing the adherence of CFSE-labeled P. gingivalis to CD45-negative RBCs after 5 min of incubation of 1 × 10 7 bacteria with whole blood cells suspended in autologous serum. M1 represents the bacterium-binding fraction of the RBCs. (B) A total of 1 × 10 7 CFSE-labeled P. gingivalis bacteria were incubated for various periods with whole blood cells suspended in autologous serum. The MFI for the binding of bacteria to RBCs are shown as medians and interquartile ranges for eight experiments.

    Journal: Infection and Immunity

    Article Title: The Atherogenic Bacterium Porphyromonas gingivalis Evades Circulating Phagocytes by Adhering to Erythrocytes ▿ Evades Circulating Phagocytes by Adhering to Erythrocytes ▿ †

    doi: 10.1128/IAI.01036-10

    Figure Lengend Snippet: Binding of P. gingivalis ATCC 33277 to human RBCs. (A) Flow cytometry histogram showing the adherence of CFSE-labeled P. gingivalis to CD45-negative RBCs after 5 min of incubation of 1 × 10 7 bacteria with whole blood cells suspended in autologous serum. M1 represents the bacterium-binding fraction of the RBCs. (B) A total of 1 × 10 7 CFSE-labeled P. gingivalis bacteria were incubated for various periods with whole blood cells suspended in autologous serum. The MFI for the binding of bacteria to RBCs are shown as medians and interquartile ranges for eight experiments.

    Article Snippet: We examined whether complement activation by P. gingivalis strain ATCC 33277 allows the bacterium to adhere to human red blood cells (RBCs) and thereby evade attack by circulating phagocytes.

    Techniques: Binding Assay, Flow Cytometry, Cytometry, Labeling, Incubation

    Activation of complement by P. gingivalis . The binding of fragments of C3 to P. gingivalis ATCC 33277 (P.g.; circles) following incubation for 30 min with sera from 10 healthy individuals was assessed by flow cytometry, using FITC-conjugated polyclonal rabbit anti-human C3d (recognizing native C3, C3b, iC3b, and C3d) as the detecting antibody. F. nucleatum (F.n.; squares) was used as a positive control. The data shown are net values after subtraction of background fluorescence observed in the presence of the same sera preheated to 56°C for 1 h to achieve complement inactivation. **, P

    Journal: Infection and Immunity

    Article Title: The Atherogenic Bacterium Porphyromonas gingivalis Evades Circulating Phagocytes by Adhering to Erythrocytes ▿ Evades Circulating Phagocytes by Adhering to Erythrocytes ▿ †

    doi: 10.1128/IAI.01036-10

    Figure Lengend Snippet: Activation of complement by P. gingivalis . The binding of fragments of C3 to P. gingivalis ATCC 33277 (P.g.; circles) following incubation for 30 min with sera from 10 healthy individuals was assessed by flow cytometry, using FITC-conjugated polyclonal rabbit anti-human C3d (recognizing native C3, C3b, iC3b, and C3d) as the detecting antibody. F. nucleatum (F.n.; squares) was used as a positive control. The data shown are net values after subtraction of background fluorescence observed in the presence of the same sera preheated to 56°C for 1 h to achieve complement inactivation. **, P

    Article Snippet: We examined whether complement activation by P. gingivalis strain ATCC 33277 allows the bacterium to adhere to human red blood cells (RBCs) and thereby evade attack by circulating phagocytes.

    Techniques: Activation Assay, Binding Assay, Incubation, Flow Cytometry, Cytometry, Positive Control, Fluorescence

    Adherence of P. gingivalis to RBCs is CR1 dependent. A total of 1 × 10 7 CFSE-labeled P. gingivalis ATCC 33277 bacteria were incubated with 50 μl PerCP-anti-CD45-labeled whole blood cells suspended in autologous serum in the presence of the anti-CR1 MAb HB8592 (IgG1 isotype) (open circles), anti-glycophorin A (anti-GPA) as an IgG1 isotype control (closed squares), or no antibody (−MAbs; closed circles). The adherence of P. gingivalis to CD45-negative RBCs under these conditions is shown as the median and range for four experiments. The P value signifies the probability for identical curves, as determined by two-way analysis of variance.

    Journal: Infection and Immunity

    Article Title: The Atherogenic Bacterium Porphyromonas gingivalis Evades Circulating Phagocytes by Adhering to Erythrocytes ▿ Evades Circulating Phagocytes by Adhering to Erythrocytes ▿ †

    doi: 10.1128/IAI.01036-10

    Figure Lengend Snippet: Adherence of P. gingivalis to RBCs is CR1 dependent. A total of 1 × 10 7 CFSE-labeled P. gingivalis ATCC 33277 bacteria were incubated with 50 μl PerCP-anti-CD45-labeled whole blood cells suspended in autologous serum in the presence of the anti-CR1 MAb HB8592 (IgG1 isotype) (open circles), anti-glycophorin A (anti-GPA) as an IgG1 isotype control (closed squares), or no antibody (−MAbs; closed circles). The adherence of P. gingivalis to CD45-negative RBCs under these conditions is shown as the median and range for four experiments. The P value signifies the probability for identical curves, as determined by two-way analysis of variance.

    Article Snippet: We examined whether complement activation by P. gingivalis strain ATCC 33277 allows the bacterium to adhere to human red blood cells (RBCs) and thereby evade attack by circulating phagocytes.

    Techniques: Labeling, Incubation

    Red blood cell-mediated restriction of uptake of P. gingivalis by peripheral leukocytes. A total of 1 × 10 7 CFSE-labeled P. gingivalis ATCC 33277 bacteria were added to cells derived from 100 μl normal whole blood (+RBCs) or from 100 μl whole blood deprived of red blood cells (−RBCs) by lysis in ammonium chloride buffer. Autologous serum was present during the entire incubation, at a concentration of 70% (vol/vol). The binding of the bacteria to CD15 + neutrophils (A), CD14 + monocytes (B), and CD19 + B cells (C) at various time points is shown, expressed as MFI values, and the corresponding ratios for the binding occurring in the presence and absence of red blood cells are also shown (D to F). Circles and error bars represent medians and interquartile ranges for eight experiments. (*), P = 0.09; *, P

    Journal: Infection and Immunity

    Article Title: The Atherogenic Bacterium Porphyromonas gingivalis Evades Circulating Phagocytes by Adhering to Erythrocytes ▿ Evades Circulating Phagocytes by Adhering to Erythrocytes ▿ †

    doi: 10.1128/IAI.01036-10

    Figure Lengend Snippet: Red blood cell-mediated restriction of uptake of P. gingivalis by peripheral leukocytes. A total of 1 × 10 7 CFSE-labeled P. gingivalis ATCC 33277 bacteria were added to cells derived from 100 μl normal whole blood (+RBCs) or from 100 μl whole blood deprived of red blood cells (−RBCs) by lysis in ammonium chloride buffer. Autologous serum was present during the entire incubation, at a concentration of 70% (vol/vol). The binding of the bacteria to CD15 + neutrophils (A), CD14 + monocytes (B), and CD19 + B cells (C) at various time points is shown, expressed as MFI values, and the corresponding ratios for the binding occurring in the presence and absence of red blood cells are also shown (D to F). Circles and error bars represent medians and interquartile ranges for eight experiments. (*), P = 0.09; *, P

    Article Snippet: We examined whether complement activation by P. gingivalis strain ATCC 33277 allows the bacterium to adhere to human red blood cells (RBCs) and thereby evade attack by circulating phagocytes.

    Techniques: Labeling, Derivative Assay, Lysis, Incubation, Concentration Assay, Binding Assay

    Comparative analysis of the penetration potential of P. gingivalis ATCC 33277 and its gingipain-null derivative mutant (KDP128) in the reconstituted basement membrane (Matrigel) model. 14 C-labeled P. gingivalis cells were placed on top of the Matrigel,

    Journal: Infection and Immunity

    Article Title: In Vitro Models of Tissue Penetration and Destruction by Porphyromonas gingivalis

    doi: 10.1128/IAI.72.8.4689-4698.2004

    Figure Lengend Snippet: Comparative analysis of the penetration potential of P. gingivalis ATCC 33277 and its gingipain-null derivative mutant (KDP128) in the reconstituted basement membrane (Matrigel) model. 14 C-labeled P. gingivalis cells were placed on top of the Matrigel,

    Article Snippet: As shown in Fig. , infiltration of the basement membrane and connective tissue by P. gingivalis ATCC 33277 cells was also observed.

    Techniques: Mutagenesis, Labeling

    Effect of P. gingivalis on EHOM structures revealed by transmission electron microscopy. Shown are ultrathin sections of the EHOM infected with P. gingivalis ATCC 33277 (10 9 cells) or P. gingivalis KDP128 (10 9 cells). Cells of P. gingivalis ATCC 33277

    Journal: Infection and Immunity

    Article Title: In Vitro Models of Tissue Penetration and Destruction by Porphyromonas gingivalis

    doi: 10.1128/IAI.72.8.4689-4698.2004

    Figure Lengend Snippet: Effect of P. gingivalis on EHOM structures revealed by transmission electron microscopy. Shown are ultrathin sections of the EHOM infected with P. gingivalis ATCC 33277 (10 9 cells) or P. gingivalis KDP128 (10 9 cells). Cells of P. gingivalis ATCC 33277

    Article Snippet: As shown in Fig. , infiltration of the basement membrane and connective tissue by P. gingivalis ATCC 33277 cells was also observed.

    Techniques: Transmission Assay, Electron Microscopy, Infection

    Degradation of Matrigel constituents by cells of P. gingivalis ATCC 33277 and KDP128, as determined by SDS-PAGE analysis and Coomassie blue staining. (A) Lanes: 1, molecular weight markers; 2, Matrigel alone; 3, Matrigel plus ATCC 33277; 4, Matrigel plus

    Journal: Infection and Immunity

    Article Title: In Vitro Models of Tissue Penetration and Destruction by Porphyromonas gingivalis

    doi: 10.1128/IAI.72.8.4689-4698.2004

    Figure Lengend Snippet: Degradation of Matrigel constituents by cells of P. gingivalis ATCC 33277 and KDP128, as determined by SDS-PAGE analysis and Coomassie blue staining. (A) Lanes: 1, molecular weight markers; 2, Matrigel alone; 3, Matrigel plus ATCC 33277; 4, Matrigel plus

    Article Snippet: As shown in Fig. , infiltration of the basement membrane and connective tissue by P. gingivalis ATCC 33277 cells was also observed.

    Techniques: SDS Page, Staining, Molecular Weight

    Macrophage migration-inhibitory factor (MIF) acted as a regulator in Porphyromonas gingivalis ( P. gingivalis )-induced intercellular adhesion molecule-1 (ICAM-1) expression. a Western blot analysis of ICAM-1 in EA.hy926 cells. b Quantitative analyses of the optical density relative to the internal reference protein GAPDH in Western blot analysis. c Quantitative Real-time PCR analysis of ICAM-1 mRNA. Infection with P. gingivalis ATCC 33277 (MOI = 100, 24 h) could induce the expression of ICAM-1 and ICAM-1 mRNA in EA.hy926 cells, this inductive effect of P. gingivalis was blocked by the MIF antagonist ISO-1. Sufficient exogenous recombinant MIF (rMIF) supplementation rescued the ICAM-1 expression. Data are presented as means ± SD from three independent experiments. * P

    Journal: BMC Microbiology

    Article Title: Porphyromonas gingivalis ATCC 33277 promotes intercellular adhesion molecule-1 expression in endothelial cells and monocyte-endothelial cell adhesion through macrophage migration inhibitory factor

    doi: 10.1186/s12866-018-1156-1

    Figure Lengend Snippet: Macrophage migration-inhibitory factor (MIF) acted as a regulator in Porphyromonas gingivalis ( P. gingivalis )-induced intercellular adhesion molecule-1 (ICAM-1) expression. a Western blot analysis of ICAM-1 in EA.hy926 cells. b Quantitative analyses of the optical density relative to the internal reference protein GAPDH in Western blot analysis. c Quantitative Real-time PCR analysis of ICAM-1 mRNA. Infection with P. gingivalis ATCC 33277 (MOI = 100, 24 h) could induce the expression of ICAM-1 and ICAM-1 mRNA in EA.hy926 cells, this inductive effect of P. gingivalis was blocked by the MIF antagonist ISO-1. Sufficient exogenous recombinant MIF (rMIF) supplementation rescued the ICAM-1 expression. Data are presented as means ± SD from three independent experiments. * P

    Article Snippet: Compared with the control level, MIF expression was increased 2.25-fold (MOI = 100) by P. gingivalis ATCC 33277 infection for 24 h ( P < 0.01).

    Techniques: Migration, Expressing, Western Blot, Real-time Polymerase Chain Reaction, Infection, Recombinant

    Macrophage migration-inhibitory factor (MIF) regulated the increased monocyte adhesion to endothelial cells infected with Porphyromonas gingivalis ( P. gingivalis ) . a THP-1 cell adhesion to EA.hy926 cells was labeled by calcein-AM and visualized. Pictures are representative fields captured by flurencence microscope (upper line) or microscope (lower line) of three independent experiments (magnification × 100). b The histogram of the evaluation of adhered THP-1 cell assessed by cell count assay. Compared with uninfected cells, P. gingivalis ATCC 33277 infection (MOI = 100, 24 h) markedly increased THP-1 cell adhesion to endothelial cells ( P

    Journal: BMC Microbiology

    Article Title: Porphyromonas gingivalis ATCC 33277 promotes intercellular adhesion molecule-1 expression in endothelial cells and monocyte-endothelial cell adhesion through macrophage migration inhibitory factor

    doi: 10.1186/s12866-018-1156-1

    Figure Lengend Snippet: Macrophage migration-inhibitory factor (MIF) regulated the increased monocyte adhesion to endothelial cells infected with Porphyromonas gingivalis ( P. gingivalis ) . a THP-1 cell adhesion to EA.hy926 cells was labeled by calcein-AM and visualized. Pictures are representative fields captured by flurencence microscope (upper line) or microscope (lower line) of three independent experiments (magnification × 100). b The histogram of the evaluation of adhered THP-1 cell assessed by cell count assay. Compared with uninfected cells, P. gingivalis ATCC 33277 infection (MOI = 100, 24 h) markedly increased THP-1 cell adhesion to endothelial cells ( P

    Article Snippet: Compared with the control level, MIF expression was increased 2.25-fold (MOI = 100) by P. gingivalis ATCC 33277 infection for 24 h ( P < 0.01).

    Techniques: Migration, Infection, Labeling, Microscopy, Cell Counting

    Porphyromonas gingivalis ( P. gingivalis ) ATCC 33277 infection enhances MIF secretion in EA.hy926 cells . EA.hy926 were challenged with Escherichia coli ( E. coli ) lipopolysaccharide (LPS, 1 μg/mL) or P. gingivalis (MOI = 100) for 4, 10 or 24 h. MIF levels was analyzed using ELISA. Infection of P. gingivalis significantly increased MIF secretion in EA.hy926 cells. EA.hy926 were incubated with medium only as a control. E.coli LPS was used as a positive control. The I bar shows the standard deviation. * P

    Journal: BMC Microbiology

    Article Title: Porphyromonas gingivalis ATCC 33277 promotes intercellular adhesion molecule-1 expression in endothelial cells and monocyte-endothelial cell adhesion through macrophage migration inhibitory factor

    doi: 10.1186/s12866-018-1156-1

    Figure Lengend Snippet: Porphyromonas gingivalis ( P. gingivalis ) ATCC 33277 infection enhances MIF secretion in EA.hy926 cells . EA.hy926 were challenged with Escherichia coli ( E. coli ) lipopolysaccharide (LPS, 1 μg/mL) or P. gingivalis (MOI = 100) for 4, 10 or 24 h. MIF levels was analyzed using ELISA. Infection of P. gingivalis significantly increased MIF secretion in EA.hy926 cells. EA.hy926 were incubated with medium only as a control. E.coli LPS was used as a positive control. The I bar shows the standard deviation. * P

    Article Snippet: Compared with the control level, MIF expression was increased 2.25-fold (MOI = 100) by P. gingivalis ATCC 33277 infection for 24 h ( P < 0.01).

    Techniques: Infection, Enzyme-linked Immunosorbent Assay, Incubation, Positive Control, Standard Deviation

    Illustration of a model for protective/destructive feedback through CD24-NLRP3 and P. gingivalis -NLRP3 interactions in healthy and diseased periodontal tissues. The non-keratinised lining epithelium (junctional epithelium) provides initial responses to bacterial products by signalling through receptors of innate immunity to activate NLRP3 inflammasome pathways (right panel). These comprise an intracellular network of regulatory and effector molecules leading to synthesis and activation of pro-inflammatory cytokines [IL-1β (green dots) IL-18 (red dots)]. However, activated IL-18 potently suppresses NLRP3 and release of IL-18. Destructive feedback accelerates periodontal tissue damage. Conversely, CD24 is characteristically strongly expressed in the lining epithelium of inflamed tissues and functions as an important negative regulator for response to danger signals, protecting tissues from excessive leukocyte activity mediated by microbial activity (left panel).

    Journal: Immunity, Inflammation and Disease

    Article Title: CD24 activates the NLRP3 inflammasome through c-Src kinase activity in a model of the lining epithelium of inflamed periodontal tissues

    doi: 10.1002/iid3.40

    Figure Lengend Snippet: Illustration of a model for protective/destructive feedback through CD24-NLRP3 and P. gingivalis -NLRP3 interactions in healthy and diseased periodontal tissues. The non-keratinised lining epithelium (junctional epithelium) provides initial responses to bacterial products by signalling through receptors of innate immunity to activate NLRP3 inflammasome pathways (right panel). These comprise an intracellular network of regulatory and effector molecules leading to synthesis and activation of pro-inflammatory cytokines [IL-1β (green dots) IL-18 (red dots)]. However, activated IL-18 potently suppresses NLRP3 and release of IL-18. Destructive feedback accelerates periodontal tissue damage. Conversely, CD24 is characteristically strongly expressed in the lining epithelium of inflamed tissues and functions as an important negative regulator for response to danger signals, protecting tissues from excessive leukocyte activity mediated by microbial activity (left panel).

    Article Snippet: RNA isolation and quantitative real-time RT-PCR for inflammasomes Confluent H413 clone-1 cells were incubated with one of the following conditions for 3 h: 5 µg/ml of CD24 mouse monoclonal (ALB9) peptide antibody (IgG1, GeneTex Inc USA), which recognises a short non-glycosylated peptide sequence close to the site of GPI linkage to the protein core of the cluster-w4/CD24 antigen ; treated with an IgG1 negative control antibody (5 µg/ml, DAKO, Denmark); treated with CD24 peptide antibody (5 µg/ml) plus c-Src inhibitor saracatinib (AZD0530, 1 µM); treated with recombinant IL-18 at 5 ng/ml in media; and treated with P. gingivalis strain (ATCC 33277) at a multiplicity of infection (MOI) of 100 .

    Techniques: Activation Assay, Activity Assay

    Quantitative real-time reverse transcription (RT)-PCR for gene expression of inflammasome and tight junction components in H413 epithelial cells in response to CD24 peptide antibody (a), active recombinant IL-18 (b and c), P. gingivalis (d) (* P

    Journal: Immunity, Inflammation and Disease

    Article Title: CD24 activates the NLRP3 inflammasome through c-Src kinase activity in a model of the lining epithelium of inflamed periodontal tissues

    doi: 10.1002/iid3.40

    Figure Lengend Snippet: Quantitative real-time reverse transcription (RT)-PCR for gene expression of inflammasome and tight junction components in H413 epithelial cells in response to CD24 peptide antibody (a), active recombinant IL-18 (b and c), P. gingivalis (d) (* P

    Article Snippet: RNA isolation and quantitative real-time RT-PCR for inflammasomes Confluent H413 clone-1 cells were incubated with one of the following conditions for 3 h: 5 µg/ml of CD24 mouse monoclonal (ALB9) peptide antibody (IgG1, GeneTex Inc USA), which recognises a short non-glycosylated peptide sequence close to the site of GPI linkage to the protein core of the cluster-w4/CD24 antigen ; treated with an IgG1 negative control antibody (5 µg/ml, DAKO, Denmark); treated with CD24 peptide antibody (5 µg/ml) plus c-Src inhibitor saracatinib (AZD0530, 1 µM); treated with recombinant IL-18 at 5 ng/ml in media; and treated with P. gingivalis strain (ATCC 33277) at a multiplicity of infection (MOI) of 100 .

    Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Recombinant

    The arrangement of porR and its neighbouring genes in P. gingivalis ATCC 33277 genome. porR (PGN_1236) and its neighbouring genes are flanked by IS5 family transposons that formed a composite transposon of 70 kbp in length. The genes that involve in biosynthesis of A-LPS are represented by yellow rectangles while the gene that does not involve is represented by brown rectangle. The genes for hypothetical proteins are represented by white rectangles. The genes for IS5 family transposases are represented by cyan rectangles. The purple triangles represented 12 bp inverted repeats that flanked the genes for IS5 family transposases. Name of proteins encoded by the genes are shown under rectangles that represented the genes. The slashes indicated gaps in the genome.

    Journal: PeerJ

    Article Title: Phylogenetic comparison between Type IX Secretion System (T9SS) protein components suggests evidence of horizontal gene transfer

    doi: 10.7717/peerj.9019

    Figure Lengend Snippet: The arrangement of porR and its neighbouring genes in P. gingivalis ATCC 33277 genome. porR (PGN_1236) and its neighbouring genes are flanked by IS5 family transposons that formed a composite transposon of 70 kbp in length. The genes that involve in biosynthesis of A-LPS are represented by yellow rectangles while the gene that does not involve is represented by brown rectangle. The genes for hypothetical proteins are represented by white rectangles. The genes for IS5 family transposases are represented by cyan rectangles. The purple triangles represented 12 bp inverted repeats that flanked the genes for IS5 family transposases. Name of proteins encoded by the genes are shown under rectangles that represented the genes. The slashes indicated gaps in the genome.

    Article Snippet: Identification of porR and its neighbouring genes’ arrangement in Porphyromonas gingivalis ATCC 33277 genome The sequence of P. gingivalis ATCC 33277 genome and annotation files of the genome were retrieved from Genbank ( ).

    Techniques:

    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

    P. gingivalis and its gingipains and fimbrial mutants upregulate ETS1 in AoSMCs. Quantitative real-time PCR results demonstrate relative transcription levels of ETS1 in AoSMCs stimulated with wild-type ATCC 33277 and W50 and the corresponding W50 gingipain

    Journal: Infection and Immunity

    Article Title: Gingipains from the Periodontal Pathogen Porphyromonas gingivalis Play a Significant Role in Regulation of Angiopoietin 1 and Angiopoietin 2 in Human Aortic Smooth Muscle Cells

    doi: 10.1128/IAI.00498-15

    Figure Lengend Snippet: P. gingivalis and its gingipains and fimbrial mutants upregulate ETS1 in AoSMCs. Quantitative real-time PCR results demonstrate relative transcription levels of ETS1 in AoSMCs stimulated with wild-type ATCC 33277 and W50 and the corresponding W50 gingipain

    Article Snippet: AoSMCs were infected with F. alocis or different strains of P. gingivalis at an MOI of 10 for 24 h and 48 h. The P. gingivalis ATCC 33277 strain significantly increased the Angpt2 protein expression in AoSMCs at 24 h after infection (see Fig. S4A and B in the supplemental material), whereas both ATCC 33277 and W50 significantly increased the Angpt2 protein level at 48 h after infection ( and ).

    Techniques: Real-time Polymerase Chain Reaction

    Schematic representation of preparation of recombinant proteins of the proteinase and adhesin domains of RgpA. The DNA for each domain of RgpA was amplified by PCR with primers (arrows) using P. gingivalis ATCC 33277 chromosomal DNA as a template. The PCR products were inserted into plasmid pET28a. The numbers in the RgpA structure indicate the predicted amino acid residues assigned based on its nucleotide sequence.

    Journal: Infection and Immunity

    Article Title: A Functional Virulence Complex Composed of Gingipains, Adhesins, and Lipopolysaccharide Shows High Affinity to Host Cells and Matrix Proteins and Escapes Recognition by Host Immune Systems

    doi: 10.1128/IAI.73.2.883-893.2005

    Figure Lengend Snippet: Schematic representation of preparation of recombinant proteins of the proteinase and adhesin domains of RgpA. The DNA for each domain of RgpA was amplified by PCR with primers (arrows) using P. gingivalis ATCC 33277 chromosomal DNA as a template. The PCR products were inserted into plasmid pET28a. The numbers in the RgpA structure indicate the predicted amino acid residues assigned based on its nucleotide sequence.

    Article Snippet: The expression plasmids were constructed as follows: DNA for each domain of RgpA was amplified by PCR with various primers using P. gingivalis ATCC 33277 chromosomal DNA as a template (Fig. ).

    Techniques: Recombinant, Amplification, Polymerase Chain Reaction, Plasmid Preparation, Sequencing

    Treatment with Porphyromonas gingivalis more effectively stimulated production of IL-1β messenger RNA (mRNA), pro-IL-1β, and IL-1β protein than Streptococcus mitis . Adenosine 5'-triphosphate (ATP) in conjunction with P. gingivalis , but not S. mitis , exhibited an enhanced signaling on the induction of IL-1β expression. (A–C) Dose-dependent (MOI = 0.5, 5, and 50) and time course (2, 6, and 24 h) assay of phorbol-12-myristate-13-acetate (PMA)-primed THP-1 cells infected with P. gingivalis or S. mitis . IL-1β mRNA expression was measured by real-time qPCR (Aa,b) , intracellular pro-IL-1β was detected by immunoblotting (B) , and mature IL-1β secreted into supernatant was assayed by ELISA (Ca,b) . (D–E) PMA-primed THP-1 cells were infected with P. gingivalis or S. mitis (MOI = 50) with or without ATP for 2 h. IL-1β mRNA expression was measured by real-time PCR (Da) , mature IL-1β secreted into supernatant was assayed by ELISA (Db) , and intracellular pro-IL-1β was detected by immunoblotting (Ea,b) . Data of real-time qPCR and ELISA represent means ± SD of at least three independent experiments and results of immunoblot analysis were representative of at least three experiments. * p

    Journal: Frontiers in Microbiology

    Article Title: Porphyromonas gingivalis-Induced NLRP3 Inflammasome Activation and Its Downstream Interleukin-1β Release Depend on Caspase-4

    doi: 10.3389/fmicb.2020.01881

    Figure Lengend Snippet: Treatment with Porphyromonas gingivalis more effectively stimulated production of IL-1β messenger RNA (mRNA), pro-IL-1β, and IL-1β protein than Streptococcus mitis . Adenosine 5'-triphosphate (ATP) in conjunction with P. gingivalis , but not S. mitis , exhibited an enhanced signaling on the induction of IL-1β expression. (A–C) Dose-dependent (MOI = 0.5, 5, and 50) and time course (2, 6, and 24 h) assay of phorbol-12-myristate-13-acetate (PMA)-primed THP-1 cells infected with P. gingivalis or S. mitis . IL-1β mRNA expression was measured by real-time qPCR (Aa,b) , intracellular pro-IL-1β was detected by immunoblotting (B) , and mature IL-1β secreted into supernatant was assayed by ELISA (Ca,b) . (D–E) PMA-primed THP-1 cells were infected with P. gingivalis or S. mitis (MOI = 50) with or without ATP for 2 h. IL-1β mRNA expression was measured by real-time PCR (Da) , mature IL-1β secreted into supernatant was assayed by ELISA (Db) , and intracellular pro-IL-1β was detected by immunoblotting (Ea,b) . Data of real-time qPCR and ELISA represent means ± SD of at least three independent experiments and results of immunoblot analysis were representative of at least three experiments. * p

    Article Snippet: Bacterial Culture P. gingivalis (ATCC 33277) was grown on brain heart infusion (BHI) agar supplemented with 5% defibrinated sheep blood containing 5 mg/ml hemin and 0.5 mg/ml vitamin K (3-phytyl-menadione) under anaerobic conditions at 37°C.

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

    Induction of P. gingivalis -specific antibodies in sera from rgpA DNA vaccine-immunized mice. Serial dilutions were used to measure the endpoint titers (≥0.2). The rgpA DNA vaccine was injected, as shown by arrows, in a total of five inoculations. The results are means ± standard deviations of log 2 ELISA antibody titers. The variation in the endpoint titers was within 1 twofold serial dilution.

    Journal: Infection and Immunity

    Article Title: Arg-Gingipain A DNA Vaccine Induces Protective Immunity against Infection by Porphyromonas gingivalis in a Murine Model

    doi: 10.1128/IAI.69.5.2858-2864.2001

    Figure Lengend Snippet: Induction of P. gingivalis -specific antibodies in sera from rgpA DNA vaccine-immunized mice. Serial dilutions were used to measure the endpoint titers (≥0.2). The rgpA DNA vaccine was injected, as shown by arrows, in a total of five inoculations. The results are means ± standard deviations of log 2 ELISA antibody titers. The variation in the endpoint titers was within 1 twofold serial dilution.

    Article Snippet: The region encoding the catalytic domain and hemagglutinin domain of Arg-gingipain A was amplified by PCR for P. gingivalis ATCC 33277 genomic DNA.

    Techniques: Mouse Assay, Injection, Enzyme-linked Immunosorbent Assay, Serial Dilution

    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

    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

    The frequency of total CD4 + T cells producing IL-17A was significantly higher in the CLN of C57BL/6J dams than in C57BL/6NCrl dams after oral inoculation with P. gingivalis . CD4 + T cells from MLN (A and B) or PaLN-CLN (C and D) of pregnant dams were stimulated with PMA-ionomycin and analyzed by flow cytometry for total expression of IL-17A or IFN-γ after P. gingivalis oral inoculation. The percentages of total CD4 + T cells producing cytokines across groups were compared using a Mann-Whitney nonparametric test, since the cytokine data did not have a normal distribution. Medians plus interquartile ranges are shown in the plot. C57BL/6J (B6/J) dams are represented by square symbols; C57BL/6NCrl (B6/NCrl, SFB + ) dams are represented by circular symbols.

    Journal: Infection and Immunity

    Article Title: Fetal Weight Outcomes in C57BL/6J and C57BL/6NCrl Mice after Oral Colonization with Porphyromonas gingivalis

    doi: 10.1128/IAI.00280-19

    Figure Lengend Snippet: The frequency of total CD4 + T cells producing IL-17A was significantly higher in the CLN of C57BL/6J dams than in C57BL/6NCrl dams after oral inoculation with P. gingivalis . CD4 + T cells from MLN (A and B) or PaLN-CLN (C and D) of pregnant dams were stimulated with PMA-ionomycin and analyzed by flow cytometry for total expression of IL-17A or IFN-γ after P. gingivalis oral inoculation. The percentages of total CD4 + T cells producing cytokines across groups were compared using a Mann-Whitney nonparametric test, since the cytokine data did not have a normal distribution. Medians plus interquartile ranges are shown in the plot. C57BL/6J (B6/J) dams are represented by square symbols; C57BL/6NCrl (B6/NCrl, SFB + ) dams are represented by circular symbols.

    Article Snippet: Given that total number of Th17 cells, but not Th1 cells, was higher in the CLN-PaLN of B6/J dams than in B6/NCrl dams after oral colonization with P. gingivalis , we further investigated the specificity of the response to determine whether there was a correlation between the frequency of P. gingivalis -specific CD4+ T cells and low fetal weight.

    Techniques: Flow Cytometry, Expressing, MANN-WHITNEY

    The fetal weight was independent of the frequency of CD4 + T cells producing IL-17A or IFN-γ in response to P. gingivalis in both substrains of C57BL/6 dams orally colonized with P. gingivalis . A Spearman correlation test was utilized to assess the relationship between the average fetal weight ± the SD and the frequency of CD4 + T cells producing IL-17A or IFN-γ in response to restimulation with P. gingivalis using ELISpot. PMA-ionomycin stimulation served as a positive control, demonstrating that CD4 + T cells were capable of cytokine production in culture (data not shown). C57BL/6J (B6/J) dams are displayed as squares, and C57BL/6NCrl (B6/NCrl) dams are displayed as circles.

    Journal: Infection and Immunity

    Article Title: Fetal Weight Outcomes in C57BL/6J and C57BL/6NCrl Mice after Oral Colonization with Porphyromonas gingivalis

    doi: 10.1128/IAI.00280-19

    Figure Lengend Snippet: The fetal weight was independent of the frequency of CD4 + T cells producing IL-17A or IFN-γ in response to P. gingivalis in both substrains of C57BL/6 dams orally colonized with P. gingivalis . A Spearman correlation test was utilized to assess the relationship between the average fetal weight ± the SD and the frequency of CD4 + T cells producing IL-17A or IFN-γ in response to restimulation with P. gingivalis using ELISpot. PMA-ionomycin stimulation served as a positive control, demonstrating that CD4 + T cells were capable of cytokine production in culture (data not shown). C57BL/6J (B6/J) dams are displayed as squares, and C57BL/6NCrl (B6/NCrl) dams are displayed as circles.

    Article Snippet: Given that total number of Th17 cells, but not Th1 cells, was higher in the CLN-PaLN of B6/J dams than in B6/NCrl dams after oral colonization with P. gingivalis , we further investigated the specificity of the response to determine whether there was a correlation between the frequency of P. gingivalis -specific CD4+ T cells and low fetal weight.

    Techniques: Enzyme-linked Immunospot, Positive Control

    The frequency of CD4 + T cells producing IL-17A or IFN-γ was significantly increased in response to oral P. gingivalis ( Pg ) in nonpregnant females from both C57BL/6 substrains. CD4 + T cells from PaLN-CLN of pregnant dams or females that had a copulation plug but no embryos on E17 were analyzed by ELISpot for the expression of IL-17A (A and B) or IFN-γ (C and D) in response to restimulation with P. gingivalis . Groups were compared using a Mann-Whitney nonparametric test, since the cytokine data did not have a normal distribution. Sham dams are represented by unshaded symbols; P. gingivalis -colonized dams are represented by shaded symbols. C57BL/6J (B6/J) dams are represented as squares, and C57BL/6NCrl (B6/NCrl) dams are represented as circles. The medians and interquartile ranges are plotted.

    Journal: Infection and Immunity

    Article Title: Fetal Weight Outcomes in C57BL/6J and C57BL/6NCrl Mice after Oral Colonization with Porphyromonas gingivalis

    doi: 10.1128/IAI.00280-19

    Figure Lengend Snippet: The frequency of CD4 + T cells producing IL-17A or IFN-γ was significantly increased in response to oral P. gingivalis ( Pg ) in nonpregnant females from both C57BL/6 substrains. CD4 + T cells from PaLN-CLN of pregnant dams or females that had a copulation plug but no embryos on E17 were analyzed by ELISpot for the expression of IL-17A (A and B) or IFN-γ (C and D) in response to restimulation with P. gingivalis . Groups were compared using a Mann-Whitney nonparametric test, since the cytokine data did not have a normal distribution. Sham dams are represented by unshaded symbols; P. gingivalis -colonized dams are represented by shaded symbols. C57BL/6J (B6/J) dams are represented as squares, and C57BL/6NCrl (B6/NCrl) dams are represented as circles. The medians and interquartile ranges are plotted.

    Article Snippet: Given that total number of Th17 cells, but not Th1 cells, was higher in the CLN-PaLN of B6/J dams than in B6/NCrl dams after oral colonization with P. gingivalis , we further investigated the specificity of the response to determine whether there was a correlation between the frequency of P. gingivalis -specific CD4+ T cells and low fetal weight.

    Techniques: Enzyme-linked Immunospot, Expressing, MANN-WHITNEY

    C57BL/6J mice experience reduced fetal weight in response to oral colonization with P. gingivalis . The average fetal weight after oral colonization with P. gingivalis in C57BL/6J (B6/J) and C57BL/6NCrl (B6/NCrl) dams was determined. Symbols show the average fetal weight in each litter per dam. B6/J dams are represented as squares and B6/NCrl dams are represented as circles. A Student unpaired t test with Welch’s correction was used to compare the average fetal weight per dam. Experimental groups shown display the average ± the standard deviation (SD).

    Journal: Infection and Immunity

    Article Title: Fetal Weight Outcomes in C57BL/6J and C57BL/6NCrl Mice after Oral Colonization with Porphyromonas gingivalis

    doi: 10.1128/IAI.00280-19

    Figure Lengend Snippet: C57BL/6J mice experience reduced fetal weight in response to oral colonization with P. gingivalis . The average fetal weight after oral colonization with P. gingivalis in C57BL/6J (B6/J) and C57BL/6NCrl (B6/NCrl) dams was determined. Symbols show the average fetal weight in each litter per dam. B6/J dams are represented as squares and B6/NCrl dams are represented as circles. A Student unpaired t test with Welch’s correction was used to compare the average fetal weight per dam. Experimental groups shown display the average ± the standard deviation (SD).

    Article Snippet: Given that total number of Th17 cells, but not Th1 cells, was higher in the CLN-PaLN of B6/J dams than in B6/NCrl dams after oral colonization with P. gingivalis , we further investigated the specificity of the response to determine whether there was a correlation between the frequency of P. gingivalis -specific CD4+ T cells and low fetal weight.

    Techniques: Mouse Assay, Standard Deviation

    Increasing amounts of P. gingivalis DNA in C57BL/6J placentas are correlated with reduced fetal weight. The percentages of placentas with (black) or without (gray) P. gingivalis DNA in C57BL/6J (B6/J, A) and C57BL/6NCrl (B6/NCrl, B) substrains were determined. The correlations between fetal weight and the amount of P. gingivalis DNA detected in murine DNA normalized to 250 ng of total DNA isolated from whole placenta of B6/J (C) and B6/NCrl (D) dams were determined. Each data point represents the weight of one fetus ( x ) in relation to amount of P. gingivalis DNA in the corresponding placenta ( y ). The limit of detection of the nested qPCR (dashed line) was 0.05 pg equating to 8 CFU. Data were converted to equivalent CFU of P. gingivalis to demonstrate clinical relevance.

    Journal: Infection and Immunity

    Article Title: Fetal Weight Outcomes in C57BL/6J and C57BL/6NCrl Mice after Oral Colonization with Porphyromonas gingivalis

    doi: 10.1128/IAI.00280-19

    Figure Lengend Snippet: Increasing amounts of P. gingivalis DNA in C57BL/6J placentas are correlated with reduced fetal weight. The percentages of placentas with (black) or without (gray) P. gingivalis DNA in C57BL/6J (B6/J, A) and C57BL/6NCrl (B6/NCrl, B) substrains were determined. The correlations between fetal weight and the amount of P. gingivalis DNA detected in murine DNA normalized to 250 ng of total DNA isolated from whole placenta of B6/J (C) and B6/NCrl (D) dams were determined. Each data point represents the weight of one fetus ( x ) in relation to amount of P. gingivalis DNA in the corresponding placenta ( y ). The limit of detection of the nested qPCR (dashed line) was 0.05 pg equating to 8 CFU. Data were converted to equivalent CFU of P. gingivalis to demonstrate clinical relevance.

    Article Snippet: Given that total number of Th17 cells, but not Th1 cells, was higher in the CLN-PaLN of B6/J dams than in B6/NCrl dams after oral colonization with P. gingivalis , we further investigated the specificity of the response to determine whether there was a correlation between the frequency of P. gingivalis -specific CD4+ T cells and low fetal weight.

    Techniques: Isolation, Real-time Polymerase Chain Reaction

    The fetal weight was independent of the frequency of total CD4 + T cells producing IL-17A or IFN-γ in both substrains of C57BL/6 dams orally colonized with P. gingivalis . CD4 + T cells from PaLN-CLN of C57BL/6J (B6/J) mice or C57BL/6NCrl (B6/NCrl) mice orally colonized with P. gingivalis were stimulated with PMA-ionomycin in the presence of brefeldin A before intracellular cytokine staining for flow cytometry. A Spearman correlation test was utilized to assess the relationship between the average fetal weight ± the SD and the frequency of total CD4 + T cells producing IL-17A or IFN-γ after oral colonization with P. gingivalis . B6/J dams are represented as squares, and B6/NCrl dams are represented as circles.

    Journal: Infection and Immunity

    Article Title: Fetal Weight Outcomes in C57BL/6J and C57BL/6NCrl Mice after Oral Colonization with Porphyromonas gingivalis

    doi: 10.1128/IAI.00280-19

    Figure Lengend Snippet: The fetal weight was independent of the frequency of total CD4 + T cells producing IL-17A or IFN-γ in both substrains of C57BL/6 dams orally colonized with P. gingivalis . CD4 + T cells from PaLN-CLN of C57BL/6J (B6/J) mice or C57BL/6NCrl (B6/NCrl) mice orally colonized with P. gingivalis were stimulated with PMA-ionomycin in the presence of brefeldin A before intracellular cytokine staining for flow cytometry. A Spearman correlation test was utilized to assess the relationship between the average fetal weight ± the SD and the frequency of total CD4 + T cells producing IL-17A or IFN-γ after oral colonization with P. gingivalis . B6/J dams are represented as squares, and B6/NCrl dams are represented as circles.

    Article Snippet: Given that total number of Th17 cells, but not Th1 cells, was higher in the CLN-PaLN of B6/J dams than in B6/NCrl dams after oral colonization with P. gingivalis , we further investigated the specificity of the response to determine whether there was a correlation between the frequency of P. gingivalis -specific CD4+ T cells and low fetal weight.

    Techniques: Mouse Assay, Staining, Flow Cytometry

    Scanning electron microscopy images of Porphyromonas gingivalis cells attached to the sandblasted large-grit acid-etched surface: (A, F) control group; (B, G) photodynamic therapy group; (C, H) brush alone group; (D, I) brushing with a LED light group; (E, J) brushing with a LED light and erythrosine group. LED: light-emitting diode.

    Journal: Journal of Periodontal & Implant Science

    Article Title: Efficacy of an LED toothbrush on a Porphyromonas gingivalis biofilm on a sandblasted and acid-etched titanium surface: an in vitro study

    doi: 10.5051/jpis.2018.48.3.164

    Figure Lengend Snippet: Scanning electron microscopy images of Porphyromonas gingivalis cells attached to the sandblasted large-grit acid-etched surface: (A, F) control group; (B, G) photodynamic therapy group; (C, H) brush alone group; (D, I) brushing with a LED light group; (E, J) brushing with a LED light and erythrosine group. LED: light-emitting diode.

    Article Snippet: P. gingivalis was grown in trypticase soy broth (Becton, Dickinson and Company, Sparks, MD, USA) containing 5 μg/mL haemin (Sigma Chemical Co., St. Louis, MO, USA) and 1 μg/mL menadione (Sigma Chemical Co.) under anaerobic conditions in an atmosphere of 5% CO2 , 5% H2 , and 90% N2 for 3–4 days.

    Techniques: Electron Microscopy

    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: For OMV preparation, P. gingivalis was grown in 250 ml tryptic soy broth (TSB) supplemented with hemin and menadione for 48 h under anaerobic conditions.

    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: For OMV preparation, P. gingivalis was grown in 250 ml tryptic soy broth (TSB) supplemented with hemin and menadione for 48 h under anaerobic conditions.

    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: For OMV preparation, P. gingivalis was grown in 250 ml tryptic soy broth (TSB) supplemented with hemin and menadione for 48 h under anaerobic conditions.

    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: For OMV preparation, P. gingivalis was grown in 250 ml tryptic soy broth (TSB) supplemented with hemin and menadione for 48 h under anaerobic conditions.

    Techniques:

    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

    Distribution of PAD in various strains of P. gingivalis . The production of citrulline from BAEE was determined in cellular (open bar), supernatant (solid bar), and vesicle (hatched bar) fractions from HG66, ATCC 53978 (W50), and ATCC 33277 strains of P. gingivalis .

    Journal: Infection and Immunity

    Article Title: Purification, Characterization, and Sequence Analysis of a Potential Virulence Factor from Porphyromonas gingivalis, Peptidylarginine Deiminase

    doi:

    Figure Lengend Snippet: Distribution of PAD in various strains of P. gingivalis . The production of citrulline from BAEE was determined in cellular (open bar), supernatant (solid bar), and vesicle (hatched bar) fractions from HG66, ATCC 53978 (W50), and ATCC 33277 strains of P. gingivalis .

    Article Snippet: Culture fluid (1,500 ml) from 36 h of cultivation of P. gingivalis ATCC 53978 (W50) and ATCC 33277 was obtained by centrifugation of cells (6,000 × g ; 30 min; 4°C).

    Techniques:

    Genomic representation of the P. gingivalis proteome, showing changes in relative abundance for the P. gingivalis - F. nucleatum - S. gordonii / P. gingivalis comparison by spectral counting . Each dot represents a PGN ORF number in the order followed by the ATCC 33277 strain annotation. Color codes: red, over-expression in the P. gingivalis - F. nucleatum - S. gordonii community relative to P. gingivalis alone; green, under-expression in the community relative to P. gingivalis alone; yellow, protein was detected qualitatively, but did not change in abundance; gray, proteins that were qualitative non-detects; gaps indicate ORFs that were not common to both the ATCC 33277 and W83 annotations according to a master cross-reference compiled by LANL (G. Xie, personal communication).

    Journal: BMC Microbiology

    Article Title: Proteomics of Porphyromonas gingivalis within a model oral microbial community

    doi: 10.1186/1471-2180-9-98

    Figure Lengend Snippet: Genomic representation of the P. gingivalis proteome, showing changes in relative abundance for the P. gingivalis - F. nucleatum - S. gordonii / P. gingivalis comparison by spectral counting . Each dot represents a PGN ORF number in the order followed by the ATCC 33277 strain annotation. Color codes: red, over-expression in the P. gingivalis - F. nucleatum - S. gordonii community relative to P. gingivalis alone; green, under-expression in the community relative to P. gingivalis alone; yellow, protein was detected qualitatively, but did not change in abundance; gray, proteins that were qualitative non-detects; gaps indicate ORFs that were not common to both the ATCC 33277 and W83 annotations according to a master cross-reference compiled by LANL (G. Xie, personal communication).

    Article Snippet: 3.1 curation of P. gingivalis W83 (2006, TIGR-CMR [ ]), S. gordonii Challis NCTC7868 (2007, TIGR-CMR [ ], F. nucleatum ATCC 25586 (2002, TIGR-CMR [ ]), bovine (2005, UC Santa Cruz), nrdb human subset (NCBI, as provided with Thermo Bioworks ver.

    Techniques: Over Expression, 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

    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:

    Quantitative and qualitative biofilm assays. (A) Quantitative analysis of biofilm accretion by P. gingivalis strains and chimeras. Chimeric strains 2, 4, and 9 (dark bars) were compared to strain 33277TcEm for statistical analysis by t test. All other chimeras (light bars) were compared to W83TcEm. Results represent three independent experiments, each done in triplicate ( n = 9). Symbols * and # represent P values of

    Journal: Journal of Bacteriology

    Article Title: Conjugal Transfer of Chromosomal DNA Contributes to Genetic Variation in the Oral Pathogen Porphyromonas gingivalis ▿

    doi: 10.1128/JB.00460-07

    Figure Lengend Snippet: Quantitative and qualitative biofilm assays. (A) Quantitative analysis of biofilm accretion by P. gingivalis strains and chimeras. Chimeric strains 2, 4, and 9 (dark bars) were compared to strain 33277TcEm for statistical analysis by t test. All other chimeras (light bars) were compared to W83TcEm. Results represent three independent experiments, each done in triplicate ( n = 9). Symbols * and # represent P values of

    Article Snippet: These results show that the exchange of genetic information between individual strains leads to measurable differences in the phenotypic behavior of P. gingivalis .

    Techniques:

    Effect of 25(OH)D 3 and 1,25(OH) 2 D 3 on the production of pro-inflammatory mediators by hPdLF with silenced VDR. Cells were transfected with either VDR siRNA or control siRNA and stimulated with P. gingivalis LPS ( Pg LPS, 1 µg/ml) or heat-killed P. gingivalis (hk Pg , 10 8 cells/ml) for 24 h in the presence or in the absence of 25(OH)D 3 or 1,25(OH) 2 D 3 . # means significantly different from control group (2 −▵▵Ct = 1). * means significantly different from cells stimulated with either P. gingivalis LPS or heat-killed P. gingivalis only

    Journal: PLoS ONE

    Article Title: Both 25-Hydroxyvitamin-D3 and 1,25-Dihydroxyvitamin-D3 Reduces Inflammatory Response in Human Periodontal Ligament Cells

    doi: 10.1371/journal.pone.0090301

    Figure Lengend Snippet: Effect of 25(OH)D 3 and 1,25(OH) 2 D 3 on the production of pro-inflammatory mediators by hPdLF with silenced VDR. Cells were transfected with either VDR siRNA or control siRNA and stimulated with P. gingivalis LPS ( Pg LPS, 1 µg/ml) or heat-killed P. gingivalis (hk Pg , 10 8 cells/ml) for 24 h in the presence or in the absence of 25(OH)D 3 or 1,25(OH) 2 D 3 . # means significantly different from control group (2 −▵▵Ct = 1). * means significantly different from cells stimulated with either P. gingivalis LPS or heat-killed P. gingivalis only

    Article Snippet: Commercially available ultrapure P. gingivalis LPS, heat-killed P. gingivalis , and ultrapure E. coli LPS (all from Invivogen, San Diego, USA) were used in the present study.

    Techniques: Transfection

    Effect of 25(OH)D 3 and 1,25(OH) 2 D 3 on the gene-expression levels of pro-inflammatory mediators in primary hPdLC in response to stimulation with P. gingivalis LPS or heat-killed P. gingivalis . Cells were stimulated with P. gingivalis LPS ( Pg LPS, 1 µg/ml) or heat-killed P. gingivalis (hk Pg , 10 8 cells/ml) for 24 h in the presence or in the absence of different concentrations of 25(OH)D 3 or 1,25(OH) 2 D 3 . Gene expression levels of IL-6 (A), IL-8 (B), and MCP-1 (C) were measured using q-PCR. Y-axes represent the n-fold expression levels of target gene in relation to non-stimulated cells (control). Data are presented as mean±SEM of six different donors. # means significantly different from control group (2 −▵▵Ct = 1). * means significantly different from cells stimulated with P. gingivalis LPS or heat-killed P. gingivalis only.

    Journal: PLoS ONE

    Article Title: Both 25-Hydroxyvitamin-D3 and 1,25-Dihydroxyvitamin-D3 Reduces Inflammatory Response in Human Periodontal Ligament Cells

    doi: 10.1371/journal.pone.0090301

    Figure Lengend Snippet: Effect of 25(OH)D 3 and 1,25(OH) 2 D 3 on the gene-expression levels of pro-inflammatory mediators in primary hPdLC in response to stimulation with P. gingivalis LPS or heat-killed P. gingivalis . Cells were stimulated with P. gingivalis LPS ( Pg LPS, 1 µg/ml) or heat-killed P. gingivalis (hk Pg , 10 8 cells/ml) for 24 h in the presence or in the absence of different concentrations of 25(OH)D 3 or 1,25(OH) 2 D 3 . Gene expression levels of IL-6 (A), IL-8 (B), and MCP-1 (C) were measured using q-PCR. Y-axes represent the n-fold expression levels of target gene in relation to non-stimulated cells (control). Data are presented as mean±SEM of six different donors. # means significantly different from control group (2 −▵▵Ct = 1). * means significantly different from cells stimulated with P. gingivalis LPS or heat-killed P. gingivalis only.

    Article Snippet: Commercially available ultrapure P. gingivalis LPS, heat-killed P. gingivalis , and ultrapure E. coli LPS (all from Invivogen, San Diego, USA) were used in the present study.

    Techniques: Expressing, Polymerase Chain Reaction

    Effect of 25(OH)D 3 and 1,25(OH) 2 D 3 on the production of pro-inflammatory mediators by hPdLF in response to stimulation with P. gingivalis LPS. Cells were stimulated with P. gingivalis LPS ( Pg LPS, 1 µg/ml) for 24 h in the presence or in the absence of different concentrations of 25(OH)D 3 or 1,25(OH) 2 D 3 . The levels of IL-6 (A), IL-8 (B), and MCP-1 (C) were measured in cell supernatants using ELISA. # means significantly different from control group (non stimulated cells). * means significantly different from group stimulated with P.gingivalis LPS only

    Journal: PLoS ONE

    Article Title: Both 25-Hydroxyvitamin-D3 and 1,25-Dihydroxyvitamin-D3 Reduces Inflammatory Response in Human Periodontal Ligament Cells

    doi: 10.1371/journal.pone.0090301

    Figure Lengend Snippet: Effect of 25(OH)D 3 and 1,25(OH) 2 D 3 on the production of pro-inflammatory mediators by hPdLF in response to stimulation with P. gingivalis LPS. Cells were stimulated with P. gingivalis LPS ( Pg LPS, 1 µg/ml) for 24 h in the presence or in the absence of different concentrations of 25(OH)D 3 or 1,25(OH) 2 D 3 . The levels of IL-6 (A), IL-8 (B), and MCP-1 (C) were measured in cell supernatants using ELISA. # means significantly different from control group (non stimulated cells). * means significantly different from group stimulated with P.gingivalis LPS only

    Article Snippet: Commercially available ultrapure P. gingivalis LPS, heat-killed P. gingivalis , and ultrapure E. coli LPS (all from Invivogen, San Diego, USA) were used in the present study.

    Techniques: Enzyme-linked Immunosorbent Assay

    Effect of 25(OH)D 3 and 1,25(OH) 2 D 3 on the gene-expression levels of pro-inflammatory mediators in hPdLF with silenced VDR in response to stimulation with P. gingivalis LPS or heat-killed P. gingivalis . Gene expression levels of IL-6, IL-8, and MCP-1 were measured using q-PCR in hPdLF after transfection with either VDR siRNA or control siRNA and stimulation with P. gingivalis LPS (A, Pg LPS, 1 µg/ml) or heat-killed P. gingivalis (B, hk Pg , 10 8 cells/ml) in the presence or in the absence of 25(OH)D 3 or 1,25(OH) 2 D 3 . Y-axes represent the n-fold expression levels of target gene in relation to non-stimulated cells. # means significantly different from control group (2 −▵▵Ct = 1). * means significantly different from cells stimulated with heat-killed P. gingivalis LPS or heat-killed P. gingivalis only.

    Journal: PLoS ONE

    Article Title: Both 25-Hydroxyvitamin-D3 and 1,25-Dihydroxyvitamin-D3 Reduces Inflammatory Response in Human Periodontal Ligament Cells

    doi: 10.1371/journal.pone.0090301

    Figure Lengend Snippet: Effect of 25(OH)D 3 and 1,25(OH) 2 D 3 on the gene-expression levels of pro-inflammatory mediators in hPdLF with silenced VDR in response to stimulation with P. gingivalis LPS or heat-killed P. gingivalis . Gene expression levels of IL-6, IL-8, and MCP-1 were measured using q-PCR in hPdLF after transfection with either VDR siRNA or control siRNA and stimulation with P. gingivalis LPS (A, Pg LPS, 1 µg/ml) or heat-killed P. gingivalis (B, hk Pg , 10 8 cells/ml) in the presence or in the absence of 25(OH)D 3 or 1,25(OH) 2 D 3 . Y-axes represent the n-fold expression levels of target gene in relation to non-stimulated cells. # means significantly different from control group (2 −▵▵Ct = 1). * means significantly different from cells stimulated with heat-killed P. gingivalis LPS or heat-killed P. gingivalis only.

    Article Snippet: Commercially available ultrapure P. gingivalis LPS, heat-killed P. gingivalis , and ultrapure E. coli LPS (all from Invivogen, San Diego, USA) were used in the present study.

    Techniques: Expressing, Polymerase Chain Reaction, Transfection

    Effect of 25(OH)D 3 and 1,25(OH) 2 D 3 on the production of pro-inflammatory mediators by primary hPdLC in response to stimulation with P. gingivalis LPS or heat-killed P. gingivalis . Cells were stimulated with P. gingivalis LPS ( Pg LPS, 1 µg/ml) or heat-killed P. gingivalis (hk Pg , 10 8 cells/ml) for 24 h in the presence or in the absence of different concentrations of 25(OH)D 3 or 1,25(OH) 2 D 3 . The levels of IL-6 (A), IL-8 (B), and MCP-1 (C) were measured in cell supernatants using ELISA. # means significantly different from control group (non stimulated cells). * means significantly different from group stimulated with P.gingivalis LPS or heat-killed P. gingivalis only.

    Journal: PLoS ONE

    Article Title: Both 25-Hydroxyvitamin-D3 and 1,25-Dihydroxyvitamin-D3 Reduces Inflammatory Response in Human Periodontal Ligament Cells

    doi: 10.1371/journal.pone.0090301

    Figure Lengend Snippet: Effect of 25(OH)D 3 and 1,25(OH) 2 D 3 on the production of pro-inflammatory mediators by primary hPdLC in response to stimulation with P. gingivalis LPS or heat-killed P. gingivalis . Cells were stimulated with P. gingivalis LPS ( Pg LPS, 1 µg/ml) or heat-killed P. gingivalis (hk Pg , 10 8 cells/ml) for 24 h in the presence or in the absence of different concentrations of 25(OH)D 3 or 1,25(OH) 2 D 3 . The levels of IL-6 (A), IL-8 (B), and MCP-1 (C) were measured in cell supernatants using ELISA. # means significantly different from control group (non stimulated cells). * means significantly different from group stimulated with P.gingivalis LPS or heat-killed P. gingivalis only.

    Article Snippet: Commercially available ultrapure P. gingivalis LPS, heat-killed P. gingivalis , and ultrapure E. coli LPS (all from Invivogen, San Diego, USA) were used in the present study.

    Techniques: Enzyme-linked Immunosorbent Assay

    Gene expression levels of pro-inflammatory mediators in hPdLF in response to stimulation with E. coli LPS, P. gingivalis LPS, and heat-killed P. gingivalis . Cells were stimulated with E. coli LPS (1 µg/ml), P. gingivalis LPS (0.1–1 µg/ml), or heat-killed P. gingivalis (10 7 –10 8 cells/ml) for 24 h. Gene expression levels of IL-6 (A), IL-8 (B), and MCP-1 (C) were measured using q-PCR. Y-axes represent the n-fold expression levels of target gene in relation to non-stimulated cells (control).

    Journal: PLoS ONE

    Article Title: Both 25-Hydroxyvitamin-D3 and 1,25-Dihydroxyvitamin-D3 Reduces Inflammatory Response in Human Periodontal Ligament Cells

    doi: 10.1371/journal.pone.0090301

    Figure Lengend Snippet: Gene expression levels of pro-inflammatory mediators in hPdLF in response to stimulation with E. coli LPS, P. gingivalis LPS, and heat-killed P. gingivalis . Cells were stimulated with E. coli LPS (1 µg/ml), P. gingivalis LPS (0.1–1 µg/ml), or heat-killed P. gingivalis (10 7 –10 8 cells/ml) for 24 h. Gene expression levels of IL-6 (A), IL-8 (B), and MCP-1 (C) were measured using q-PCR. Y-axes represent the n-fold expression levels of target gene in relation to non-stimulated cells (control).

    Article Snippet: Commercially available ultrapure P. gingivalis LPS, heat-killed P. gingivalis , and ultrapure E. coli LPS (all from Invivogen, San Diego, USA) were used in the present study.

    Techniques: Expressing, Polymerase Chain Reaction

    Effect of 25(OH)D 3 and 1,25(OH) 2 D 3 on the gene-expression levels of pro-inflammatory mediators in hPdLF in response to stimulation with P. gingivalis LPS. Cells were stimulated with P. gingivalis LPS ( Pg LPS, 1 µg/ml) for 24 h in the presence or in the absence of different concentrations of 25(OH)D 3 or 1,25(OH) 2 D 3 . Gene expression levels of IL-6 (A), IL-8 (B), and MCP-1 (C) were measured using q-PCR. Y-axes represent the n-fold expression levels of target gene in relation to non-stimulated cells (control). # means significantly different from control group (2 −▵▵Ct = 1). * means significantly different from cells stimulated with P. gingivalis LPS only.

    Journal: PLoS ONE

    Article Title: Both 25-Hydroxyvitamin-D3 and 1,25-Dihydroxyvitamin-D3 Reduces Inflammatory Response in Human Periodontal Ligament Cells

    doi: 10.1371/journal.pone.0090301

    Figure Lengend Snippet: Effect of 25(OH)D 3 and 1,25(OH) 2 D 3 on the gene-expression levels of pro-inflammatory mediators in hPdLF in response to stimulation with P. gingivalis LPS. Cells were stimulated with P. gingivalis LPS ( Pg LPS, 1 µg/ml) for 24 h in the presence or in the absence of different concentrations of 25(OH)D 3 or 1,25(OH) 2 D 3 . Gene expression levels of IL-6 (A), IL-8 (B), and MCP-1 (C) were measured using q-PCR. Y-axes represent the n-fold expression levels of target gene in relation to non-stimulated cells (control). # means significantly different from control group (2 −▵▵Ct = 1). * means significantly different from cells stimulated with P. gingivalis LPS only.

    Article Snippet: Commercially available ultrapure P. gingivalis LPS, heat-killed P. gingivalis , and ultrapure E. coli LPS (all from Invivogen, San Diego, USA) were used in the present study.

    Techniques: Expressing, Polymerase Chain Reaction