p. gingivalis strains Search Results


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  • 99
    ATCC p gingivalis strains
    PPAD supplementation restores the ability of P. <t>gingivalis</t> ΔPPAD to adhere to and invade primary human gingival fibroblasts (PHGF)
    P Gingivalis Strains, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 312 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher p gingivalis strains
    Citrullination of CXCL8 by different clinical isolates of P. <t>gingivalis</t> . Bacterial suspensions with starting concentrations of 5 × 10 7 CFU/ml were first grown for 24 h at 37°C in an anaerobic atmosphere. Then, the strains were treated
    P Gingivalis Strains, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 42 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Nissui Pharmaceutical p gingivalis strains
    Citrullination of CXCL8 by different clinical isolates of P. <t>gingivalis</t> . Bacterial suspensions with starting concentrations of 5 × 10 7 CFU/ml were first grown for 24 h at 37°C in an anaerobic atmosphere. Then, the strains were treated
    P Gingivalis Strains, supplied by Nissui Pharmaceutical, used in various techniques. Bioz Stars score: 86/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    ATCC p gingivalis strain atcc 53978
    Citrullination of CXCL8 by different clinical isolates of P. <t>gingivalis</t> . Bacterial suspensions with starting concentrations of 5 × 10 7 CFU/ml were first grown for 24 h at 37°C in an anaerobic atmosphere. Then, the strains were treated
    P Gingivalis Strain Atcc 53978, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    Becton Dickinson p gingivalis strain
    Circular map of the chromosome of P. <t>gingivalis</t> strain ATCC 33277. From the outside, the first and second circles show CDSs on the plus and minus strands, respectively. CDSs conserved in strains ATCC 33277 and W83 are indicated in red and ATCC 33277-specific CDSs in blue. The 3rd to 5th circles show IS elements (orange, IS Pg1 ; light green, IS Pg2 ; magenta, IS Pg3 ; cyan, IS Pg4 ; brown, IS Pg5 ; blue, IS Pg6 ), MITEs (magenta, MITE239; black, MITE PgRS ; cyan, MITE700), CTns, and Tns (blue, CTnPg1-a, CTnPg1-b, CTnPg2, and CTnPg3; red, Tn Pg17 ), respectively. The 6th and 7th circles show rrn operons and tRNA genes, respectively. The 8th circle shows the result of χ 2 analysis of nucleotide composition. Regions exhibiting values of > 600 are indicated in red and those of
    P Gingivalis Strain, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Becton Dickinson p gingivalis strain 381
    Circular map of the chromosome of P. <t>gingivalis</t> strain ATCC 33277. From the outside, the first and second circles show CDSs on the plus and minus strands, respectively. CDSs conserved in strains ATCC 33277 and W83 are indicated in red and ATCC 33277-specific CDSs in blue. The 3rd to 5th circles show IS elements (orange, IS Pg1 ; light green, IS Pg2 ; magenta, IS Pg3 ; cyan, IS Pg4 ; brown, IS Pg5 ; blue, IS Pg6 ), MITEs (magenta, MITE239; black, MITE PgRS ; cyan, MITE700), CTns, and Tns (blue, CTnPg1-a, CTnPg1-b, CTnPg2, and CTnPg3; red, Tn Pg17 ), respectively. The 6th and 7th circles show rrn operons and tRNA genes, respectively. The 8th circle shows the result of χ 2 analysis of nucleotide composition. Regions exhibiting values of > 600 are indicated in red and those of
    P Gingivalis Strain 381, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 90/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Difco p gingivalis strain 381
    Inactivation of PG1828 in P. <t>gingivalis</t> . (A) Construction of a PG1828-deficient mutant of P. gingivalis <t>strain</t> 381 by allelic exchange mutagenesis with an ermF-ermAM antibiotic resistance cassette (as described in Materials and Methods). (B) Predicted maps of the genomes of the wild-type and the PG1828-deficient mutant. Arrowheads indicate the numbers and positions of oligonucleotide primers for PCR analysis (as described in Materials and Methods). (C) PCR analysis of WT and ΔPG1828. Numbers above the lanes indicate the primer pairs (B) used for the PCR analysis. Lane M, DNA marker; left margin, molecular sizes.
    P Gingivalis Strain 381, supplied by Difco, used in various techniques. Bioz Stars score: 86/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Strategic BioSolutions Inc p gingivalis strain w83
    Representative microscopic images of P. <t>gingivalis</t> strain <t>W83</t> within LC3 positive or LAMP-1 positive vacuoles at 6 hours post-inoculation. Arrows indicate magnified inserts. Scale bar is equivalent to 10 µm. Representative images of other P. gingivalis strains can be viewed in supporting file Figure S4 .
    P Gingivalis Strain W83, supplied by Strategic BioSolutions Inc, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Thermo Fisher p gingivalis wild type strain 381
    Representative microscopic images of P. <t>gingivalis</t> strain <t>W83</t> within LC3 positive or LAMP-1 positive vacuoles at 6 hours post-inoculation. Arrows indicate magnified inserts. Scale bar is equivalent to 10 µm. Representative images of other P. gingivalis strains can be viewed in supporting file Figure S4 .
    P Gingivalis Wild Type Strain 381, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Bio-Rad p gingivalis strain w83 competent cells
    Effect of Lys 129 mutation on Kgp expression and activity. Western blotting analysis of parental strain P. <t>gingivalis</t> <t>W83</t> and mutants K129A, K129E, and K129R is shown. Late exponential/early stationary bacterial cultures (BC) were separated by centrifugation into cell-free growth medium (GM) and cell pellet. The latter was washed, giving rise to the whole cell (WC) fraction, which was further fractionated into the soluble intracellular protein fraction (CP + PP) and the cell envelope fracion (CE) by sonication and ultracentrifugation. A and B , all fractions were standardized to the initial volume of the culture subjected to centrifugation and analyzed by Western blotting to detect Kgp forms ( A ) and Rgps (control) ( B ). C and D , Kgp ( C ) and Rgp ( D ) gingipain activities determined in whole cultures, cell-free growth medium, and washed cells using specific substrates. The whole-culture activity of the wild-type strain was arbitrarily taken as 100%.
    P Gingivalis Strain W83 Competent Cells, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 93/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Becton Dickinson pro atherogenic treatment p gingivalis strain 381
    Effect of Lys 129 mutation on Kgp expression and activity. Western blotting analysis of parental strain P. <t>gingivalis</t> <t>W83</t> and mutants K129A, K129E, and K129R is shown. Late exponential/early stationary bacterial cultures (BC) were separated by centrifugation into cell-free growth medium (GM) and cell pellet. The latter was washed, giving rise to the whole cell (WC) fraction, which was further fractionated into the soluble intracellular protein fraction (CP + PP) and the cell envelope fracion (CE) by sonication and ultracentrifugation. A and B , all fractions were standardized to the initial volume of the culture subjected to centrifugation and analyzed by Western blotting to detect Kgp forms ( A ) and Rgps (control) ( B ). C and D , Kgp ( C ) and Rgp ( D ) gingipain activities determined in whole cultures, cell-free growth medium, and washed cells using specific substrates. The whole-culture activity of the wild-type strain was arbitrarily taken as 100%.
    Pro Atherogenic Treatment P Gingivalis Strain 381, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 86/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    ATCC p gingivalis strain w83
    Effect of Lys 129 mutation on Kgp expression and activity. Western blotting analysis of parental strain P. <t>gingivalis</t> <t>W83</t> and mutants K129A, K129E, and K129R is shown. Late exponential/early stationary bacterial cultures (BC) were separated by centrifugation into cell-free growth medium (GM) and cell pellet. The latter was washed, giving rise to the whole cell (WC) fraction, which was further fractionated into the soluble intracellular protein fraction (CP + PP) and the cell envelope fracion (CE) by sonication and ultracentrifugation. A and B , all fractions were standardized to the initial volume of the culture subjected to centrifugation and analyzed by Western blotting to detect Kgp forms ( A ) and Rgps (control) ( B ). C and D , Kgp ( C ) and Rgp ( D ) gingipain activities determined in whole cultures, cell-free growth medium, and washed cells using specific substrates. The whole-culture activity of the wild-type strain was arbitrarily taken as 100%.
    P Gingivalis Strain W83, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC p gingivalis strain fdc381
    Effect of Lys 129 mutation on Kgp expression and activity. Western blotting analysis of parental strain P. <t>gingivalis</t> <t>W83</t> and mutants K129A, K129E, and K129R is shown. Late exponential/early stationary bacterial cultures (BC) were separated by centrifugation into cell-free growth medium (GM) and cell pellet. The latter was washed, giving rise to the whole cell (WC) fraction, which was further fractionated into the soluble intracellular protein fraction (CP + PP) and the cell envelope fracion (CE) by sonication and ultracentrifugation. A and B , all fractions were standardized to the initial volume of the culture subjected to centrifugation and analyzed by Western blotting to detect Kgp forms ( A ) and Rgps (control) ( B ). C and D , Kgp ( C ) and Rgp ( D ) gingipain activities determined in whole cultures, cell-free growth medium, and washed cells using specific substrates. The whole-culture activity of the wild-type strain was arbitrarily taken as 100%.
    P Gingivalis Strain Fdc381, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Nissui Pharmaceutical p gingivalis
    P. <t>gingivalis</t> enhances IL-1β secretion and caspase-1 activation, leading to inflammatory cell death. (A) PMA-primed THP-1 cells were infected with P. gingivalis (MOI, 100) for 6 or 24 h. Cell culture supernatants were assayed for human IL-1β
    P Gingivalis, supplied by Nissui Pharmaceutical, used in various techniques. Bioz Stars score: 90/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Thermo Fisher p gingivalis
    P. <t>gingivalis</t> enhances IL-1β secretion and caspase-1 activation, leading to inflammatory cell death. (A) PMA-primed THP-1 cells were infected with P. gingivalis (MOI, 100) for 6 or 24 h. Cell culture supernatants were assayed for human IL-1β
    P Gingivalis, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 369 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    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: 94/100, based on 97 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Difco p gingivalis
    Electron micrograph showing changes in surface structures of P. <t>gingivalis</t> ATCC 33277 and W83. Bacterial cells grown to the log phase (OD 600 of 0.7–0.9) were processed for electron microscopic examination using formvar-carbon coated grids (500 mesh) and were examined using Philips Tecnai 12 TEM. Fimbriae were lacking in the vimF mutant FLL476 when compared with the wild ATCC33277 and the complemented strain FLL476C’. A thick glycocalyx along with vesicles and a well-defined outer membrane was observed in W83. FLL95 showed hazy outer membrane with reduced visicles. In the complemented strain FLL95C’ the outer membrane morphology was restored.
    P Gingivalis, supplied by Difco, used in various techniques. Bioz Stars score: 89/100, based on 52 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson p gingivalis
    Electron micrograph showing changes in surface structures of P. <t>gingivalis</t> ATCC 33277 and W83. Bacterial cells grown to the log phase (OD 600 of 0.7–0.9) were processed for electron microscopic examination using formvar-carbon coated grids (500 mesh) and were examined using Philips Tecnai 12 TEM. Fimbriae were lacking in the vimF mutant FLL476 when compared with the wild ATCC33277 and the complemented strain FLL476C’. A thick glycocalyx along with vesicles and a well-defined outer membrane was observed in W83. FLL95 showed hazy outer membrane with reduced visicles. In the complemented strain FLL95C’ the outer membrane morphology was restored.
    P Gingivalis, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 90 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    InvivoGen p gingivalis
    Proliferation and differentiation of CD4 + T cells mediated by gingipains from P. <t>gingivalis</t> . Monocyte-derived dendritic cells (moDC) were untreated or exposed to P. gingivalis W83 in the presence/absence of specific protease inhibitors (KYT-1 and KYT-36, each at a concentration of 1 μM) or the isogenic gingipain-null mutant ΔKΔRAB for 6 h at MOI (multiplicity of infection) 1:50. Naive CD4 + T cells stained with CFSE were added and co-stimulated for 5 days. Proliferation of lymphocytes was measured at day 1, 3, and 5. (A) Completed results of proliferation on day 1, 3, and 5 as a percentage of CFSE low cells. Data are presented as a mean ± standard deviation of assays performed in triplicate using four independent donors, and were analyzed with a two way ANOVA with the Bonferoni's posttest correction (# P
    P Gingivalis, supplied by InvivoGen, used in various techniques. Bioz Stars score: 95/100, based on 317 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    InvivoGen p gingivalis lps
    IRAK1 expression was inhibited by miR-146a in B cells after challenged with P. <t>gingivalis</t> <t>LPS</t> miR-146a mimic could reduce the mRNA and protein levels of IRAK1 as detected by PCR (A) and ELISA (B). Also, addition of miR-146a inhibited TRAF6 expression in B cells at both mRNA (C) and protein level (D). Inhibition of miR-146a by its inhibitor significantly elevated both mRNA and protein levels of IRAK1 expression in B cells (E and F), as well as mRNA (G) and protein (H) levels of TRAF6 expression in B cells. The experiments were performed in triplicate. n=6–8. Data are presented as mean±SD. One-way ANOVA. * p
    P Gingivalis Lps, supplied by InvivoGen, used in various techniques. Bioz Stars score: 95/100, based on 154 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    ATCC lyophilized p gingivalis
    IRAK1 expression was inhibited by miR-146a in B cells after challenged with P. <t>gingivalis</t> <t>LPS</t> miR-146a mimic could reduce the mRNA and protein levels of IRAK1 as detected by PCR (A) and ELISA (B). Also, addition of miR-146a inhibited TRAF6 expression in B cells at both mRNA (C) and protein level (D). Inhibition of miR-146a by its inhibitor significantly elevated both mRNA and protein levels of IRAK1 expression in B cells (E and F), as well as mRNA (G) and protein (H) levels of TRAF6 expression in B cells. The experiments were performed in triplicate. n=6–8. Data are presented as mean±SD. One-way ANOVA. * p
    Lyophilized P Gingivalis, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


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

    Journal: Molecular oral microbiology

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

    doi: 10.1111/omi.12081

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

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

    Techniques:

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

    Journal: Molecular oral microbiology

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

    doi: 10.1111/omi.12081

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

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

    Techniques:

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

    Journal: Molecular oral microbiology

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

    doi: 10.1111/omi.12081

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

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

    Techniques:

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

    Journal: Infection and Immunity

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

    doi: 10.1128/IAI.01013-06

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

    Article Snippet: Our results demonstrate that not only WT P. gingivalis strains with various virulences (ATCC 33277, 381, and W83) but also KDP136 directly enter the endocytic pathway to lysosomes in the infected HAECs, but also that their trafficking to the autophagosome is minor.

    Techniques: Infection

    Sequencing quality control and reproducibility. Panel A shows quality scores of the Illumina sequencing reads for mapping. Fifty base pair single-end reads were obtained with ‘high’ quality out to ~42 base pairs and ‘good’ quality out to ~47 base pairs. The green background corresponds to high quality reads, the yellow background to intermediate quality reads and the red background to poor quality reads. Data shown are for the number of high, intermediate and low quality reads at a specific number of base pairs away from the transposon. The yellow bar encompasses the 25-75 th percentile and the red horizontal bar indicates the mean. The green bracket identifies the base pair position where high quality reads comprise the over 75% of the total reads. Blue arrow signifies where typical amount of sequencing that can be obtained when preparing DNA using the MmeI restriction site, demonstrating superior mapping and analysis ability of C-tailing method. No sequencing reads shorter than 20 bp were used for analyses. B ) Replicates of the same library were sequenced in separate experiments. The graph compares the number of insertions per gene for technical replicates 1 and 2 of P. gingivalis strain ATCC 33277 Mariner mutant library and showed excellent correlation between the replicates (R 2 =0.9892). Median number of insertions when excluding genes containing zero is 9 while the mean is 17. Sixteen genes have 100 insertions or greater.

    Journal: BMC Genomics

    Article Title: Identification of essential genes of the periodontal pathogen Porphyromonas gingivalis

    doi: 10.1186/1471-2164-13-578

    Figure Lengend Snippet: Sequencing quality control and reproducibility. Panel A shows quality scores of the Illumina sequencing reads for mapping. Fifty base pair single-end reads were obtained with ‘high’ quality out to ~42 base pairs and ‘good’ quality out to ~47 base pairs. The green background corresponds to high quality reads, the yellow background to intermediate quality reads and the red background to poor quality reads. Data shown are for the number of high, intermediate and low quality reads at a specific number of base pairs away from the transposon. The yellow bar encompasses the 25-75 th percentile and the red horizontal bar indicates the mean. The green bracket identifies the base pair position where high quality reads comprise the over 75% of the total reads. Blue arrow signifies where typical amount of sequencing that can be obtained when preparing DNA using the MmeI restriction site, demonstrating superior mapping and analysis ability of C-tailing method. No sequencing reads shorter than 20 bp were used for analyses. B ) Replicates of the same library were sequenced in separate experiments. The graph compares the number of insertions per gene for technical replicates 1 and 2 of P. gingivalis strain ATCC 33277 Mariner mutant library and showed excellent correlation between the replicates (R 2 =0.9892). Median number of insertions when excluding genes containing zero is 9 while the mean is 17. Sixteen genes have 100 insertions or greater.

    Article Snippet: Identifying putative essential genes of P. gingivalis by Tn-seq The genome of P. gingivalis strain ATCC 33277 comprises 2.35 Mb of chromosomal DNA and no plasmids.

    Techniques: Sequencing, Mutagenesis

    Mapping of the P. gingivalis essential genes. In blue in the outermost ring are shown the putative P. gingivalis essential genes identified by transposon mutagenesis of strain ATCC 33277. Blue arrows depict the orientation of the essential genes. Genetic loci for positive strand (outer set of arrows) are shown in the outermost circle and genetic loci for the negative strand are shown in the innermost ring. In red tick marks are coding sequences for P. gingivalis strain W83, in green are coding sequences for strain W50 and in blue are coding sequences for strain TDC60. Shaded black area represents GC-content for given regions. CGViewer ( http://stothard.afns.ualberta.ca/cgview_server/ ) was utilized to visualize the entire circular genome of P. gingivalis strain ATCC 33277 with the putative essential genes labeled [ 52 ]. NCBI FASTA files of P. gingivalis strains W83, W50 and TDC60 were used for BLAST matching to the ATCC 33277 base genome.

    Journal: BMC Genomics

    Article Title: Identification of essential genes of the periodontal pathogen Porphyromonas gingivalis

    doi: 10.1186/1471-2164-13-578

    Figure Lengend Snippet: Mapping of the P. gingivalis essential genes. In blue in the outermost ring are shown the putative P. gingivalis essential genes identified by transposon mutagenesis of strain ATCC 33277. Blue arrows depict the orientation of the essential genes. Genetic loci for positive strand (outer set of arrows) are shown in the outermost circle and genetic loci for the negative strand are shown in the innermost ring. In red tick marks are coding sequences for P. gingivalis strain W83, in green are coding sequences for strain W50 and in blue are coding sequences for strain TDC60. Shaded black area represents GC-content for given regions. CGViewer ( http://stothard.afns.ualberta.ca/cgview_server/ ) was utilized to visualize the entire circular genome of P. gingivalis strain ATCC 33277 with the putative essential genes labeled [ 52 ]. NCBI FASTA files of P. gingivalis strains W83, W50 and TDC60 were used for BLAST matching to the ATCC 33277 base genome.

    Article Snippet: Identifying putative essential genes of P. gingivalis by Tn-seq The genome of P. gingivalis strain ATCC 33277 comprises 2.35 Mb of chromosomal DNA and no plasmids.

    Techniques: Mutagenesis, Labeling

    Distribution of P. gingivalis ATCC 33277 essential genes by Cluster of Orthologous Groups (COG) classifications. A ) Number of genes within a COG category; essential genes in blue and entire genome in red. B ) Percent of essential genes within a COG category from total number in genome; red line represents the 22% that 463/2102 of the percent essential over the entire genome. [A = RNA processing and modification, B = Chromatin structure and dynamics, C = Energy production and conversion, D = Cell cycle control, E = Amino acid metabolism and transport, F = Nucleotide metabolism and transport, G = Carbohydrate metabolism and transport, H = Coenzyme metabolism, I = Lipid metabolism, J = Translation, K = Transcription, L = Replication and repair, M = Cell wall/membrane/envelop biogenesis, N = Cell motility, O = Post-translational modification, protein turnover, chaperone functions, P = Inorganic ion transport and metabolism, Q = Secondary structure, T = Signal transduction, U = Intracellular trafficking and secretion, Y = Nuclear structure, Z = Cytoskeleton, R = General functional prediction only, S = Function unknown].

    Journal: BMC Genomics

    Article Title: Identification of essential genes of the periodontal pathogen Porphyromonas gingivalis

    doi: 10.1186/1471-2164-13-578

    Figure Lengend Snippet: Distribution of P. gingivalis ATCC 33277 essential genes by Cluster of Orthologous Groups (COG) classifications. A ) Number of genes within a COG category; essential genes in blue and entire genome in red. B ) Percent of essential genes within a COG category from total number in genome; red line represents the 22% that 463/2102 of the percent essential over the entire genome. [A = RNA processing and modification, B = Chromatin structure and dynamics, C = Energy production and conversion, D = Cell cycle control, E = Amino acid metabolism and transport, F = Nucleotide metabolism and transport, G = Carbohydrate metabolism and transport, H = Coenzyme metabolism, I = Lipid metabolism, J = Translation, K = Transcription, L = Replication and repair, M = Cell wall/membrane/envelop biogenesis, N = Cell motility, O = Post-translational modification, protein turnover, chaperone functions, P = Inorganic ion transport and metabolism, Q = Secondary structure, T = Signal transduction, U = Intracellular trafficking and secretion, Y = Nuclear structure, Z = Cytoskeleton, R = General functional prediction only, S = Function unknown].

    Article Snippet: Identifying putative essential genes of P. gingivalis by Tn-seq The genome of P. gingivalis strain ATCC 33277 comprises 2.35 Mb of chromosomal DNA and no plasmids.

    Techniques: Modification, Transduction, Functional Assay

    Determination of proper transposon insertion. Confirmation of transposon insertions was performed by PCR for presence of transposon ( ermG ) ( A ). “Mwt” = molecular weight marker, “+” = positive control (gDNA from E. coli /pSAM_Bt); “-“= a negative control ( P. gingivalis ATCC 33277). All other lanes contain amplicons from PCR of individual colonies of transformed P. gingivalis . Panels ( B ) and ( C ) show PCR of the same samples using primers for the bla and himar1c19a genes respectively that are present in the plasmid, but which should be lost with proper insertion of the transposon. These three panels are a combination of separate gels; all which were run using identical PCR gDNA template for each of the separate reactions. ( D ) Nested semi-random PCR for individual mutant sequencing preparation. PCR from individual colonies was performed using primers to Mariner transposon and random primers ARB1 and ARB2 (Additional file 6 : Table S6). Two rounds of nested PCR were performed. Negative controls of wild-type P. gingivalis strain ATCC 33277 ( Pg ), template only (T) and primer only (P) lanes precede thirteen individual mutants.

    Journal: BMC Genomics

    Article Title: Identification of essential genes of the periodontal pathogen Porphyromonas gingivalis

    doi: 10.1186/1471-2164-13-578

    Figure Lengend Snippet: Determination of proper transposon insertion. Confirmation of transposon insertions was performed by PCR for presence of transposon ( ermG ) ( A ). “Mwt” = molecular weight marker, “+” = positive control (gDNA from E. coli /pSAM_Bt); “-“= a negative control ( P. gingivalis ATCC 33277). All other lanes contain amplicons from PCR of individual colonies of transformed P. gingivalis . Panels ( B ) and ( C ) show PCR of the same samples using primers for the bla and himar1c19a genes respectively that are present in the plasmid, but which should be lost with proper insertion of the transposon. These three panels are a combination of separate gels; all which were run using identical PCR gDNA template for each of the separate reactions. ( D ) Nested semi-random PCR for individual mutant sequencing preparation. PCR from individual colonies was performed using primers to Mariner transposon and random primers ARB1 and ARB2 (Additional file 6 : Table S6). Two rounds of nested PCR were performed. Negative controls of wild-type P. gingivalis strain ATCC 33277 ( Pg ), template only (T) and primer only (P) lanes precede thirteen individual mutants.

    Article Snippet: Identifying putative essential genes of P. gingivalis by Tn-seq The genome of P. gingivalis strain ATCC 33277 comprises 2.35 Mb of chromosomal DNA and no plasmids.

    Techniques: Polymerase Chain Reaction, Molecular Weight, Marker, Positive Control, Negative Control, Transformation Assay, Plasmid Preparation, Mutagenesis, Sequencing, Nested PCR

    Fimbriae are not involved in P. gingivalis -mediated regulation of Angpt1 and Angpt2 production in AoSMCs. Quantitative real-time PCR results demonstrate relative transcription levels for Angpt1 (A) and Angpt2 (B) in AoSMCs stimulated with wild-type P.

    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: Fimbriae are not involved in P. gingivalis -mediated regulation of Angpt1 and Angpt2 production in AoSMCs. Quantitative real-time PCR results demonstrate relative transcription levels for Angpt1 (A) and Angpt2 (B) in AoSMCs stimulated with wild-type P.

    Article Snippet: The wild-type P. gingivalis strains ATCC 33277 and W50 significantly increased ETS1 expression in AoSMCs after 16 and 24 h ( ).

    Techniques: Real-time Polymerase Chain Reaction

    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: The wild-type P. gingivalis strains ATCC 33277 and W50 significantly increased ETS1 expression in AoSMCs after 16 and 24 h ( ).

    Techniques: Real-time Polymerase Chain Reaction

    P. gingivalis and its gingipains regulate Angpt1 and Angpt2 expression in AoSMCs. (A and B) Quantitative real-time PCR results demonstrate relative transcription levels for Angpt1 (A) and Angpt2 (B) in AoSMCs stimulated with wild-type P. gingivalis (ATCC

    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 regulate Angpt1 and Angpt2 expression in AoSMCs. (A and B) Quantitative real-time PCR results demonstrate relative transcription levels for Angpt1 (A) and Angpt2 (B) in AoSMCs stimulated with wild-type P. gingivalis (ATCC

    Article Snippet: The wild-type P. gingivalis strains ATCC 33277 and W50 significantly increased ETS1 expression in AoSMCs after 16 and 24 h ( ).

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    Mean relative closure (±SEM) from all biological replicates of scratches in oral epithelial cells challenged with heat inactivated and viable P. gingivalis wild type strain ATCC 33277, ΔKgp, and ΔRgpArgpB mutants (a) and heat-inactivated and viable P. gingivalis wild-type strain W50, ΔKgp, and ΔRgpArgpB mutants and with addition of KYT-36 and KYT-1 peptides (b). *=significantly different from wild-type strain of the same MOI ( p

    Journal: Journal of Oral Microbiology

    Article Title: The impact of virulence factors of Porphyromonas gingivalis on wound healing in vitro

    doi: 10.3402/jom.v7.27543

    Figure Lengend Snippet: Mean relative closure (±SEM) from all biological replicates of scratches in oral epithelial cells challenged with heat inactivated and viable P. gingivalis wild type strain ATCC 33277, ΔKgp, and ΔRgpArgpB mutants (a) and heat-inactivated and viable P. gingivalis wild-type strain W50, ΔKgp, and ΔRgpArgpB mutants and with addition of KYT-36 and KYT-1 peptides (b). *=significantly different from wild-type strain of the same MOI ( p

    Article Snippet: The heat-inactivated P. gingivalis strains ATCC 33277, W83, and W50 significantly inhibited the migration of oral epithelial cells at all tested MOIs compared to control in a dose-responsive manner, although the inhibition by the heat-inactivated W83 at MOI 10 did not reach statistical significance ( ).

    Techniques:

    Mean relative closure (±SEM) from all biological replicates of scratches in oral epithelial cells challenged with LPS from P. gingivalis and LPS inactivated P. gingivalis by polymyxin B (a) and heat-inactivated P. gingivalis strains W83 and EpsC mutant (b). * p

    Journal: Journal of Oral Microbiology

    Article Title: The impact of virulence factors of Porphyromonas gingivalis on wound healing in vitro

    doi: 10.3402/jom.v7.27543

    Figure Lengend Snippet: Mean relative closure (±SEM) from all biological replicates of scratches in oral epithelial cells challenged with LPS from P. gingivalis and LPS inactivated P. gingivalis by polymyxin B (a) and heat-inactivated P. gingivalis strains W83 and EpsC mutant (b). * p

    Article Snippet: The heat-inactivated P. gingivalis strains ATCC 33277, W83, and W50 significantly inhibited the migration of oral epithelial cells at all tested MOIs compared to control in a dose-responsive manner, although the inhibition by the heat-inactivated W83 at MOI 10 did not reach statistical significance ( ).

    Techniques: Mutagenesis

    Percentage closure of a scratch in oral epithelial cells challenged with P. gingivalis ATCC 33277 and medium only.

    Journal: Journal of Oral Microbiology

    Article Title: The impact of virulence factors of Porphyromonas gingivalis on wound healing in vitro

    doi: 10.3402/jom.v7.27543

    Figure Lengend Snippet: Percentage closure of a scratch in oral epithelial cells challenged with P. gingivalis ATCC 33277 and medium only.

    Article Snippet: The heat-inactivated P. gingivalis strains ATCC 33277, W83, and W50 significantly inhibited the migration of oral epithelial cells at all tested MOIs compared to control in a dose-responsive manner, although the inhibition by the heat-inactivated W83 at MOI 10 did not reach statistical significance ( ).

    Techniques:

    Mean relative closure (±SEM) from all biological replicates of scratches in oral epithelial cells challenged with heat-inactivated and viable P. gingivalis strains ATCC 33277, W83, and W50. a=significantly different from control ( p

    Journal: Journal of Oral Microbiology

    Article Title: The impact of virulence factors of Porphyromonas gingivalis on wound healing in vitro

    doi: 10.3402/jom.v7.27543

    Figure Lengend Snippet: Mean relative closure (±SEM) from all biological replicates of scratches in oral epithelial cells challenged with heat-inactivated and viable P. gingivalis strains ATCC 33277, W83, and W50. a=significantly different from control ( p

    Article Snippet: The heat-inactivated P. gingivalis strains ATCC 33277, W83, and W50 significantly inhibited the migration of oral epithelial cells at all tested MOIs compared to control in a dose-responsive manner, although the inhibition by the heat-inactivated W83 at MOI 10 did not reach statistical significance ( ).

    Techniques:

    Effects of wortmannin and bafilomycin A 1 on viability of intracellular P. gingivalis ATCC 33277 in HAECs. (A) SDS-PAGE and immunoblot analysis of the intracellular protein levels of cathepsins B and D in uninfected and infected HAECs with ATCC 33277 or

    Journal: Infection and Immunity

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

    doi: 10.1128/IAI.01013-06

    Figure Lengend Snippet: Effects of wortmannin and bafilomycin A 1 on viability of intracellular P. gingivalis ATCC 33277 in HAECs. (A) SDS-PAGE and immunoblot analysis of the intracellular protein levels of cathepsins B and D in uninfected and infected HAECs with ATCC 33277 or

    Article Snippet: HAECs (1 × 105 cells per well in six-well tissue culture dishes) were infected with P. gingivalis WT strains (ATCC 33277 and 381) or KDP136 for 20 min at an MOI of 104 .

    Techniques: SDS Page, Infection

    Ultrastructural observations of P. gingivalis ATCC 33277- and KDP136-infected HAECs by thin-section electron microscopy. (A) At 2 h postinfection, most of the internalized ATCC 33277 organisms were enclosed by or fused with single or double membranous

    Journal: Infection and Immunity

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

    doi: 10.1128/IAI.01013-06

    Figure Lengend Snippet: Ultrastructural observations of P. gingivalis ATCC 33277- and KDP136-infected HAECs by thin-section electron microscopy. (A) At 2 h postinfection, most of the internalized ATCC 33277 organisms were enclosed by or fused with single or double membranous

    Article Snippet: HAECs (1 × 105 cells per well in six-well tissue culture dishes) were infected with P. gingivalis WT strains (ATCC 33277 and 381) or KDP136 for 20 min at an MOI of 104 .

    Techniques: Infection, Electron Microscopy

    Confocal microscopic images of intracellular P. gingivalis WT strains and KDP136 in infected cells. After infection with P. gingivalis (Pg) ATCC 33277 (A), 381 (B), W83 (C), and KDP136 (D) for 20 min, HAECs were washed and further cultured for the indicated

    Journal: Infection and Immunity

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

    doi: 10.1128/IAI.01013-06

    Figure Lengend Snippet: Confocal microscopic images of intracellular P. gingivalis WT strains and KDP136 in infected cells. After infection with P. gingivalis (Pg) ATCC 33277 (A), 381 (B), W83 (C), and KDP136 (D) for 20 min, HAECs were washed and further cultured for the indicated

    Article Snippet: HAECs (1 × 105 cells per well in six-well tissue culture dishes) were infected with P. gingivalis WT strains (ATCC 33277 and 381) or KDP136 for 20 min at an MOI of 104 .

    Techniques: Infection, Cell Culture

    Induction of autophagy in HAECs infected with P. gingivalis ATCC 33277 and KDP136. (A) Effect of starvation on LC3-staining patterns in HAECs. While the cells showed mainly diffuse staining for LC3, cells under starvation conditions for 2 h exhibited

    Journal: Infection and Immunity

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

    doi: 10.1128/IAI.01013-06

    Figure Lengend Snippet: Induction of autophagy in HAECs infected with P. gingivalis ATCC 33277 and KDP136. (A) Effect of starvation on LC3-staining patterns in HAECs. While the cells showed mainly diffuse staining for LC3, cells under starvation conditions for 2 h exhibited

    Article Snippet: HAECs (1 × 105 cells per well in six-well tissue culture dishes) were infected with P. gingivalis WT strains (ATCC 33277 and 381) or KDP136 for 20 min at an MOI of 104 .

    Techniques: Infection, Staining

    Immunoelectron microscopy of P. gingivalis -infected HAECs. After infection with ATCC 33277 and KDP136, HAECs were incubated in the absence of extracellular bacteria for the indicated times. The localization of cathepsin B, a lysosomal marker, was examined

    Journal: Infection and Immunity

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

    doi: 10.1128/IAI.01013-06

    Figure Lengend Snippet: Immunoelectron microscopy of P. gingivalis -infected HAECs. After infection with ATCC 33277 and KDP136, HAECs were incubated in the absence of extracellular bacteria for the indicated times. The localization of cathepsin B, a lysosomal marker, was examined

    Article Snippet: HAECs (1 × 105 cells per well in six-well tissue culture dishes) were infected with P. gingivalis WT strains (ATCC 33277 and 381) or KDP136 for 20 min at an MOI of 104 .

    Techniques: Immuno-Electron Microscopy, Infection, Incubation, Marker

    Correlation between duplicate samples in HCN offline headspace measurements of P. gingivalis strains. One data point (▪) represents the HCN concentrations obtained from duplicate No. 1 (horizontal axis) and No. 2 (vertical axis). Each strain was measured at 24, 48 and 72 hours, hence, there were three data points for each strain, and a total of 12 points for all four strains. A strong correlation ( r s = 0.96 and p

    Journal: Scientific Reports

    Article Title: Detection of hydrogen cyanide from oral anaerobes by cavity ring down spectroscopy

    doi: 10.1038/srep22577

    Figure Lengend Snippet: Correlation between duplicate samples in HCN offline headspace measurements of P. gingivalis strains. One data point (▪) represents the HCN concentrations obtained from duplicate No. 1 (horizontal axis) and No. 2 (vertical axis). Each strain was measured at 24, 48 and 72 hours, hence, there were three data points for each strain, and a total of 12 points for all four strains. A strong correlation ( r s = 0.96 and p

    Article Snippet: Further investigations were conducted on the species of P. gingivalis and we successfully detected HCN production (0.9–10.9 ppb) in the headspace of three P. gingivalis reference strains (ATCC 33277, W50 and OMG 434) and one clinical isolate.

    Techniques:

    Detection of HCN and CO 2 from P. gingivalis strains and blank Brucella blood agar. The P. gingivalis strains included three reference strains ( a ) ATCC 33277, ( b ) W50, ( c ) OMG 434 and one clinical isolate ( d ) 4753E. Blank Brucella blood agar ( e ) served as background control. There were duplicate samples for each strain. The detection was conducted at 24, 48 and 72 hours.

    Journal: Scientific Reports

    Article Title: Detection of hydrogen cyanide from oral anaerobes by cavity ring down spectroscopy

    doi: 10.1038/srep22577

    Figure Lengend Snippet: Detection of HCN and CO 2 from P. gingivalis strains and blank Brucella blood agar. The P. gingivalis strains included three reference strains ( a ) ATCC 33277, ( b ) W50, ( c ) OMG 434 and one clinical isolate ( d ) 4753E. Blank Brucella blood agar ( e ) served as background control. There were duplicate samples for each strain. The detection was conducted at 24, 48 and 72 hours.

    Article Snippet: Further investigations were conducted on the species of P. gingivalis and we successfully detected HCN production (0.9–10.9 ppb) in the headspace of three P. gingivalis reference strains (ATCC 33277, W50 and OMG 434) and one clinical isolate.

    Techniques:

    Dynamic profiles of HCN production by different P. gingivalis strains. The HCN concentrations from three reference strains (ATCC 33277, W50 and OMG 434) and one clinical isolate (4753E) of P. gingivalis were measured. The HCN concentrations from each strain were measured every 20 minutes. For clarity, data points are shown only every two hours.

    Journal: Scientific Reports

    Article Title: Detection of hydrogen cyanide from oral anaerobes by cavity ring down spectroscopy

    doi: 10.1038/srep22577

    Figure Lengend Snippet: Dynamic profiles of HCN production by different P. gingivalis strains. The HCN concentrations from three reference strains (ATCC 33277, W50 and OMG 434) and one clinical isolate (4753E) of P. gingivalis were measured. The HCN concentrations from each strain were measured every 20 minutes. For clarity, data points are shown only every two hours.

    Article Snippet: Further investigations were conducted on the species of P. gingivalis and we successfully detected HCN production (0.9–10.9 ppb) in the headspace of three P. gingivalis reference strains (ATCC 33277, W50 and OMG 434) and one clinical isolate.

    Techniques:

    Correlation between HCN and CO 2 concentrations of P. gingivalis ATCC 33277 and W50. Data points (▪) represent the concentration of HCN (horizontal axis) and CO 2 (vertical axis) determined from one plate at a certain time point. Since each strain was measured in duplicate and at 24, 48 and 72 hours, each strain has six data points. In total, there are 12 data points for two reference strains, P. gingivalis ATCC 33277 and W50. A strong correlation ( r s = 0.89 and p

    Journal: Scientific Reports

    Article Title: Detection of hydrogen cyanide from oral anaerobes by cavity ring down spectroscopy

    doi: 10.1038/srep22577

    Figure Lengend Snippet: Correlation between HCN and CO 2 concentrations of P. gingivalis ATCC 33277 and W50. Data points (▪) represent the concentration of HCN (horizontal axis) and CO 2 (vertical axis) determined from one plate at a certain time point. Since each strain was measured in duplicate and at 24, 48 and 72 hours, each strain has six data points. In total, there are 12 data points for two reference strains, P. gingivalis ATCC 33277 and W50. A strong correlation ( r s = 0.89 and p

    Article Snippet: Further investigations were conducted on the species of P. gingivalis and we successfully detected HCN production (0.9–10.9 ppb) in the headspace of three P. gingivalis reference strains (ATCC 33277, W50 and OMG 434) and one clinical isolate.

    Techniques: Concentration Assay

    Distinction of PPAD sorting types I and II P. gingivalis isolates were cultured for four days in BHI medium. Subsequently, bacterial cells were separated from the growth medium, and growth medium fractions, containing OMVs, were used for immunoblotting with PPAD-specific antibodies. (A) P. gingivalis reference strain W83 and the isogenic PPAD deletion mutant. (B) P. gingivalis clinical isolates. Names of sorting type II isolates are underlined. Molecular weights of marker proteins and different PPAD species are indicated.

    Journal: Virulence

    Article Title: There's no place like OM: Vesicular sorting and secretion of the peptidylarginine deiminase of Porphyromonas gingivalis

    doi: 10.1080/21505594.2017.1421827

    Figure Lengend Snippet: Distinction of PPAD sorting types I and II P. gingivalis isolates were cultured for four days in BHI medium. Subsequently, bacterial cells were separated from the growth medium, and growth medium fractions, containing OMVs, were used for immunoblotting with PPAD-specific antibodies. (A) P. gingivalis reference strain W83 and the isogenic PPAD deletion mutant. (B) P. gingivalis clinical isolates. Names of sorting type II isolates are underlined. Molecular weights of marker proteins and different PPAD species are indicated.

    Article Snippet: Additionally, the study isolates included one P. gingivalis isolate from a healthy carrier, and two P. gingivalis type strains (W83 and ATCC 33277), with the respective engineered PPAD deletion mutants [ ].

    Techniques: Cell Culture, Mutagenesis, Marker

    OMV formation by sorting type I and II isolates of P. gingivalis Electron micrographs of vesiculating cells of (A) the P. gingivalis type strain W83, and (B) the sorting type II isolate MDS33. Electron micrographs of OMVs collected from (C) strain W83, (D) the sorting type I isolate 505700, and the sorting type II isolates (E) 505759 and (F) 512915.

    Journal: Virulence

    Article Title: There's no place like OM: Vesicular sorting and secretion of the peptidylarginine deiminase of Porphyromonas gingivalis

    doi: 10.1080/21505594.2017.1421827

    Figure Lengend Snippet: OMV formation by sorting type I and II isolates of P. gingivalis Electron micrographs of vesiculating cells of (A) the P. gingivalis type strain W83, and (B) the sorting type II isolate MDS33. Electron micrographs of OMVs collected from (C) strain W83, (D) the sorting type I isolate 505700, and the sorting type II isolates (E) 505759 and (F) 512915.

    Article Snippet: Additionally, the study isolates included one P. gingivalis isolate from a healthy carrier, and two P. gingivalis type strains (W83 and ATCC 33277), with the respective engineered PPAD deletion mutants [ ].

    Techniques:

    Position of amino acid substitutions in PPAD and their impact on the electrostatic potential of the protein (A) 3D-structural ribbon representation of the PPAD protein from P. gingivalis reference strain W83, showing in yellow surface-exposed amino acid residues that have been substituted in PPAD proteins from other P. gingivalis sorting type II isolates. Catalytic residues of PPAD are shown in green. (B and C) Electrostatic potential maps showing, respectively, the PPAD proteins of strain W83 and the sorting type II isolate MDS33 from the same perspective. The two maps display the difference in electrostatic potential (red represents -5 KT/e and blue +5 KT/e ), and the respective Gln or Lys residues at position 373 are indicated. (D) 3D-structural ribbon representation of the PPAD protein from P. gingivalis strain W83, showing surface-exposed amino acid residues that have been substituted in other PPAD proteins (marked in yellow) of sorting type II isolates from the perspective of the catalytic site. The catalytic residues are marked in green. (E) Electrostatic potential map of the PPAD protein of strain W83, displaying all the residues subject to substitutions in sorting type II isolates. (F) Electrostatic potential map of the PPAD protein from the sorting type II isolate MDS33.

    Journal: Virulence

    Article Title: There's no place like OM: Vesicular sorting and secretion of the peptidylarginine deiminase of Porphyromonas gingivalis

    doi: 10.1080/21505594.2017.1421827

    Figure Lengend Snippet: Position of amino acid substitutions in PPAD and their impact on the electrostatic potential of the protein (A) 3D-structural ribbon representation of the PPAD protein from P. gingivalis reference strain W83, showing in yellow surface-exposed amino acid residues that have been substituted in PPAD proteins from other P. gingivalis sorting type II isolates. Catalytic residues of PPAD are shown in green. (B and C) Electrostatic potential maps showing, respectively, the PPAD proteins of strain W83 and the sorting type II isolate MDS33 from the same perspective. The two maps display the difference in electrostatic potential (red represents -5 KT/e and blue +5 KT/e ), and the respective Gln or Lys residues at position 373 are indicated. (D) 3D-structural ribbon representation of the PPAD protein from P. gingivalis strain W83, showing surface-exposed amino acid residues that have been substituted in other PPAD proteins (marked in yellow) of sorting type II isolates from the perspective of the catalytic site. The catalytic residues are marked in green. (E) Electrostatic potential map of the PPAD protein of strain W83, displaying all the residues subject to substitutions in sorting type II isolates. (F) Electrostatic potential map of the PPAD protein from the sorting type II isolate MDS33.

    Article Snippet: Additionally, the study isolates included one P. gingivalis isolate from a healthy carrier, and two P. gingivalis type strains (W83 and ATCC 33277), with the respective engineered PPAD deletion mutants [ ].

    Techniques:

    Map of relative abundance trends based on the ATCC 33277 gene order and annotation . This plot shows the entire set of consensus calls given in Additional file 1 : Table S1 arranged by ascending PGN number [ 11 ], which follows the physical order of genes in the genome sequence. Color coding: red indicates increased relative protein abundance for internalized P. gingivalis , green decreased relative abundance, grey indicates qualitative non-detects and black indicates an unused ORF number.

    Journal: BMC Microbiology

    Article Title: Pathway analysis for intracellular Porphyromonas gingivalis using a strain ATCC 33277 specific database

    doi: 10.1186/1471-2180-9-185

    Figure Lengend Snippet: Map of relative abundance trends based on the ATCC 33277 gene order and annotation . This plot shows the entire set of consensus calls given in Additional file 1 : Table S1 arranged by ascending PGN number [ 11 ], which follows the physical order of genes in the genome sequence. Color coding: red indicates increased relative protein abundance for internalized P. gingivalis , green decreased relative abundance, grey indicates qualitative non-detects and black indicates an unused ORF number.

    Article Snippet: Re-analysis using the P. gingivalis strain ATCC 33277 genome annotation The proteomics data previously analyzed using the strain W83 genome annotation [GenBank: AE015924 ] [ ] was recalculated employing the strain specific P. gingivalis strain ATCC 33277 annotation [GenBank: AP009380 ].

    Techniques: Sequencing

    Sensitivity of P. gingivalis mutants to air (A) and air with hydrogen peroxide (B). P. gingivalis cells that had been anaerobically cultured in enriched BHI medium overnight were diluted twice with fresh enriched BHI medium (A) or enriched BHI medium containing hydrogen peroxide (final concentration, 0.5 mM) (B) and then incubated aerobically with vigorous shaking. Samples were withdrawn at intervals and plated, after dilution in enriched BHI medium, on enriched TS plates. The plates were incubated anaerobically at 37°C for 7 days. Symbols: ○, ATCC 33277 (wild type); ▵, KDP139 ( ftn ); •, KDP141 ( dps ); ▴, KDP142 ( ftn dps ).

    Journal: Infection and Immunity

    Article Title: Purification, Gene Cloning, Gene Expression, and Mutants of Dps from the Obligate Anaerobe Porphyromonas gingivalis

    doi: 10.1128/IAI.71.3.1170-1178.2003

    Figure Lengend Snippet: Sensitivity of P. gingivalis mutants to air (A) and air with hydrogen peroxide (B). P. gingivalis cells that had been anaerobically cultured in enriched BHI medium overnight were diluted twice with fresh enriched BHI medium (A) or enriched BHI medium containing hydrogen peroxide (final concentration, 0.5 mM) (B) and then incubated aerobically with vigorous shaking. Samples were withdrawn at intervals and plated, after dilution in enriched BHI medium, on enriched TS plates. The plates were incubated anaerobically at 37°C for 7 days. Symbols: ○, ATCC 33277 (wild type); ▵, KDP139 ( ftn ); •, KDP141 ( dps ); ▴, KDP142 ( ftn dps ).

    Article Snippet: The dps gene DNA disrupted by insertion of the tetQ cartridge DNA was introduced into cells of P. gingivalis wild-type strain (ATCC 33277) and the ftn mutant (KDP139) by electroporation.

    Techniques: Cell Culture, Concentration Assay, Incubation

    Sensitivity of P. gingivalis cells to hydrogen peroxide, mitomycin C, and metronidazole. P. gingivalis ATCC 33277 (wild type), KDP139 ( ftn ), KDP141 ( dps ), and KDP142 ( ftn dps ) were anaerobically grown in enriched BHI medium for 48 h. The cells were spread on enriched TS plates, and a paper disk containing hydrogen peroxide (A), mitomycin C (B), or metronidazole (C) was placed at the centers of the plates, followed by incubation at 37°C anaerobically for 7 days. The diameters of the clear zones next to the disks were measured (in millimeters). The data shown are the means and SD of triplicate experiments.

    Journal: Infection and Immunity

    Article Title: Purification, Gene Cloning, Gene Expression, and Mutants of Dps from the Obligate Anaerobe Porphyromonas gingivalis

    doi: 10.1128/IAI.71.3.1170-1178.2003

    Figure Lengend Snippet: Sensitivity of P. gingivalis cells to hydrogen peroxide, mitomycin C, and metronidazole. P. gingivalis ATCC 33277 (wild type), KDP139 ( ftn ), KDP141 ( dps ), and KDP142 ( ftn dps ) were anaerobically grown in enriched BHI medium for 48 h. The cells were spread on enriched TS plates, and a paper disk containing hydrogen peroxide (A), mitomycin C (B), or metronidazole (C) was placed at the centers of the plates, followed by incubation at 37°C anaerobically for 7 days. The diameters of the clear zones next to the disks were measured (in millimeters). The data shown are the means and SD of triplicate experiments.

    Article Snippet: The dps gene DNA disrupted by insertion of the tetQ cartridge DNA was introduced into cells of P. gingivalis wild-type strain (ATCC 33277) and the ftn mutant (KDP139) by electroporation.

    Techniques: Incubation

    DNA-binding activity of P. gingivalis Dps. Recombinant P. gingivalis Dps purified from the E. coli overexpressing P. gingivalis dps or the recombinant P. gingivalis Dps treated with ferrous ammonium sulfate was incubated with linear DNA (1-kb DNA ladder) at 4°C for 1 h. The mixture was then subjected to agarose gel electrophoresis. DNA on the gel was stained with ethidium bromide. Lanes: 1, DNA (500 ng) alone; 2, recombinant Dps (10 μg) alone; 3, recombinant Dps (10 μg) and DNA (500 ng); 4, iron-loaded recombinant Dps (10 μg) and DNA (500 ng); 5, iron-loaded recombinant Dps (10 μg) alone.

    Journal: Infection and Immunity

    Article Title: Purification, Gene Cloning, Gene Expression, and Mutants of Dps from the Obligate Anaerobe Porphyromonas gingivalis

    doi: 10.1128/IAI.71.3.1170-1178.2003

    Figure Lengend Snippet: DNA-binding activity of P. gingivalis Dps. Recombinant P. gingivalis Dps purified from the E. coli overexpressing P. gingivalis dps or the recombinant P. gingivalis Dps treated with ferrous ammonium sulfate was incubated with linear DNA (1-kb DNA ladder) at 4°C for 1 h. The mixture was then subjected to agarose gel electrophoresis. DNA on the gel was stained with ethidium bromide. Lanes: 1, DNA (500 ng) alone; 2, recombinant Dps (10 μg) alone; 3, recombinant Dps (10 μg) and DNA (500 ng); 4, iron-loaded recombinant Dps (10 μg) and DNA (500 ng); 5, iron-loaded recombinant Dps (10 μg) alone.

    Article Snippet: The dps gene DNA disrupted by insertion of the tetQ cartridge DNA was introduced into cells of P. gingivalis wild-type strain (ATCC 33277) and the ftn mutant (KDP139) by electroporation.

    Techniques: Binding Assay, Activity Assay, Recombinant, Purification, Incubation, Agarose Gel Electrophoresis, Staining

    Alignment of the amino acid sequence of P. gingivalis Dps with those of the Dps previously isolated and determined in other prokaryotes.

    Journal: Infection and Immunity

    Article Title: Purification, Gene Cloning, Gene Expression, and Mutants of Dps from the Obligate Anaerobe Porphyromonas gingivalis

    doi: 10.1128/IAI.71.3.1170-1178.2003

    Figure Lengend Snippet: Alignment of the amino acid sequence of P. gingivalis Dps with those of the Dps previously isolated and determined in other prokaryotes.

    Article Snippet: The dps gene DNA disrupted by insertion of the tetQ cartridge DNA was introduced into cells of P. gingivalis wild-type strain (ATCC 33277) and the ftn mutant (KDP139) by electroporation.

    Techniques: Sequencing, Isolation

    Time course of induction of β-galactosidase activity in the dps′-′lacZ fusion strains. P. gingivalis KDP146 ( dps + dps′-′lacZ ) (circles) and KDP148 ( oxyR dps + dps′-′lacZ ) (triangles) were grown anaerobically in enriched BHI medium at 37°C. At an A 600 ). The β-galactosidase activity of the wild-type parent strain ATCC 33277 was

    Journal: Infection and Immunity

    Article Title: Purification, Gene Cloning, Gene Expression, and Mutants of Dps from the Obligate Anaerobe Porphyromonas gingivalis

    doi: 10.1128/IAI.71.3.1170-1178.2003

    Figure Lengend Snippet: Time course of induction of β-galactosidase activity in the dps′-′lacZ fusion strains. P. gingivalis KDP146 ( dps + dps′-′lacZ ) (circles) and KDP148 ( oxyR dps + dps′-′lacZ ) (triangles) were grown anaerobically in enriched BHI medium at 37°C. At an A 600 ). The β-galactosidase activity of the wild-type parent strain ATCC 33277 was

    Article Snippet: The dps gene DNA disrupted by insertion of the tetQ cartridge DNA was introduced into cells of P. gingivalis wild-type strain (ATCC 33277) and the ftn mutant (KDP139) by electroporation.

    Techniques: Activity Assay

    Proof of authenticity of the P. gingivalis dps mutant KDP141 and the ftn dps double mutant KDP142. (A and B) Southern blot analyses of the chromosomal DNA. The chromosomal DNAs of the wild-type ATCC 33277 (lane 1) and the ftn mutants KDP139 (lane 2), KDP141 (lane 3), and KDP142 (lane 4) were digested with Nco I. The resulting DNA fragments were subjected to agarose gel electrophresis, followed by blotting. Hybridization was performed by using the 0.6-kb Nde I- Bgl II fragment of pKD390 as a dps probe (A) and the 2.7-kb Bam HI- Bgl II fragment of pKD375 as a tetQ probe (B). (C) Immunoblot analysis. After purified P. gingivalis Dps (lane 1) and the cell extracts of ATCC 33277 (lane 2), KDP141 (lane 3), and KDP142 (lane 4) were electrophoresed through an SDS-polyacrylamide gel, the proteins were transferred to a nitrocellulose membrane and immunoreacted with antiserum against P. gingivalis Dps.

    Journal: Infection and Immunity

    Article Title: Purification, Gene Cloning, Gene Expression, and Mutants of Dps from the Obligate Anaerobe Porphyromonas gingivalis

    doi: 10.1128/IAI.71.3.1170-1178.2003

    Figure Lengend Snippet: Proof of authenticity of the P. gingivalis dps mutant KDP141 and the ftn dps double mutant KDP142. (A and B) Southern blot analyses of the chromosomal DNA. The chromosomal DNAs of the wild-type ATCC 33277 (lane 1) and the ftn mutants KDP139 (lane 2), KDP141 (lane 3), and KDP142 (lane 4) were digested with Nco I. The resulting DNA fragments were subjected to agarose gel electrophresis, followed by blotting. Hybridization was performed by using the 0.6-kb Nde I- Bgl II fragment of pKD390 as a dps probe (A) and the 2.7-kb Bam HI- Bgl II fragment of pKD375 as a tetQ probe (B). (C) Immunoblot analysis. After purified P. gingivalis Dps (lane 1) and the cell extracts of ATCC 33277 (lane 2), KDP141 (lane 3), and KDP142 (lane 4) were electrophoresed through an SDS-polyacrylamide gel, the proteins were transferred to a nitrocellulose membrane and immunoreacted with antiserum against P. gingivalis Dps.

    Article Snippet: The dps gene DNA disrupted by insertion of the tetQ cartridge DNA was introduced into cells of P. gingivalis wild-type strain (ATCC 33277) and the ftn mutant (KDP139) by electroporation.

    Techniques: Mutagenesis, Southern Blot, Agarose Gel Electrophoresis, Hybridization, Purification

    Survival of P. gingivalis cells in HUVEC. Monolayers of HUVEC were infected by P. gingivalis ATCC 33277 (wild type) and KDP141 ( dps ). Experiments were done at least five times, and the data are presented as the mean and the SD of the CFU per HUVEC.

    Journal: Infection and Immunity

    Article Title: Purification, Gene Cloning, Gene Expression, and Mutants of Dps from the Obligate Anaerobe Porphyromonas gingivalis

    doi: 10.1128/IAI.71.3.1170-1178.2003

    Figure Lengend Snippet: Survival of P. gingivalis cells in HUVEC. Monolayers of HUVEC were infected by P. gingivalis ATCC 33277 (wild type) and KDP141 ( dps ). Experiments were done at least five times, and the data are presented as the mean and the SD of the CFU per HUVEC.

    Article Snippet: The dps gene DNA disrupted by insertion of the tetQ cartridge DNA was introduced into cells of P. gingivalis wild-type strain (ATCC 33277) and the ftn mutant (KDP139) by electroporation.

    Techniques: Infection

    Differential gene expression by comparative microarray analyses (represented in log 10 ) when comparing planktonic Porphyromonas gingivalis ATCC 33277 cells either in presence of a growing biofilm or in absence of a biofilm. Control planktonic cell gene expression (X-axis) is plotted against test cells (Y-axis) with a 1.5 fold change (up or down) and p -value

    Journal: BMC Microbiology

    Article Title: Comparative gene expression analysis of planktonic Porphyromonas gingivalis ATCC 33277 in the presence of a growing biofilm versus planktonic cells

    doi: 10.1186/s12866-019-1423-9

    Figure Lengend Snippet: Differential gene expression by comparative microarray analyses (represented in log 10 ) when comparing planktonic Porphyromonas gingivalis ATCC 33277 cells either in presence of a growing biofilm or in absence of a biofilm. Control planktonic cell gene expression (X-axis) is plotted against test cells (Y-axis) with a 1.5 fold change (up or down) and p -value

    Article Snippet: Slides specific for the strain P. gingivalis ATCC 33277 [Agilent Oligo Microarrays 8x15K (074976)] were used.

    Techniques: Expressing, Microarray

    Overview of experimental design. Porphyromonas gingivalis ATCC 33277 strain was maintained on blood agar plates and grown in modified Brain Heart Infusion (BHI) medium. After 24 h, optical density was measured, and a pure culture containing 10 8 colony forming units per milliliter (CFU/mL) was set. Two culture conditions were then prepared: Test cells, depositing P. gingivalis cells in presence of Hydroxyapatite (HA) disc, and Control cells, depositing P. gingivalis cells in the wells without HA discs. After 96 h of incubation of multi-well plates under anaerobic conditions, free floating P. gingivalis cells from both test and control condition were harvested, examined by CLSM, processed and total RNA extracted and purified. Agilent Oligo Microarrays 8x15K (074976) for P. gingivalis ATCC 33277 were used for hybridizations, (this slides also contents probes against the whole genome of P. gingivalis W83), and RT-qPCR analyses were performed to confirm the results. The experiments were repeated three times and each experimental condition was pooled into the three biological replicates and processed for the transcriptomic analysis. (Images for Fig. 1 were taken from https://smart.servier.com/ under a creative commons licence)

    Journal: BMC Microbiology

    Article Title: Comparative gene expression analysis of planktonic Porphyromonas gingivalis ATCC 33277 in the presence of a growing biofilm versus planktonic cells

    doi: 10.1186/s12866-019-1423-9

    Figure Lengend Snippet: Overview of experimental design. Porphyromonas gingivalis ATCC 33277 strain was maintained on blood agar plates and grown in modified Brain Heart Infusion (BHI) medium. After 24 h, optical density was measured, and a pure culture containing 10 8 colony forming units per milliliter (CFU/mL) was set. Two culture conditions were then prepared: Test cells, depositing P. gingivalis cells in presence of Hydroxyapatite (HA) disc, and Control cells, depositing P. gingivalis cells in the wells without HA discs. After 96 h of incubation of multi-well plates under anaerobic conditions, free floating P. gingivalis cells from both test and control condition were harvested, examined by CLSM, processed and total RNA extracted and purified. Agilent Oligo Microarrays 8x15K (074976) for P. gingivalis ATCC 33277 were used for hybridizations, (this slides also contents probes against the whole genome of P. gingivalis W83), and RT-qPCR analyses were performed to confirm the results. The experiments were repeated three times and each experimental condition was pooled into the three biological replicates and processed for the transcriptomic analysis. (Images for Fig. 1 were taken from https://smart.servier.com/ under a creative commons licence)

    Article Snippet: Slides specific for the strain P. gingivalis ATCC 33277 [Agilent Oligo Microarrays 8x15K (074976)] were used.

    Techniques: Modification, Incubation, Confocal Laser Scanning Microscopy, Purification, Quantitative RT-PCR

    Effect of DNase I and proteinase K on the biofilm formation (time zero) and on 48-h-old biofilm. Fusobacterium nucleatum type strain ATCC 25586 (F.n) and Porphyromonas gingivalis type strain ATCC 33277 (P.g 33277) bacterial species were tested when they were grown as monoculture or as dual species culture. (A) DNase I effect on biofilm formation, (B) DNase I effect on 48-h biofilm, (C) Proteinase K effect on biofilm formation, (D) Proteinase K effect on 48-h biofilm. The colored columns refer to the enzyme concentrations (0.125, 0.25, 0.5, and 1 mg/ml). The y-axis represents absorbance at 570 nm. The bars represent the means with standard deviations for 3–5 samples.

    Journal: Journal of Oral Microbiology

    Article Title: Characterization of extracellular polymeric matrix, and treatment of Fusobacterium nucleatum and Porphyromonas gingivalis biofilms with DNase I and proteinase K

    doi: 10.3402/jom.v5i0.20015

    Figure Lengend Snippet: Effect of DNase I and proteinase K on the biofilm formation (time zero) and on 48-h-old biofilm. Fusobacterium nucleatum type strain ATCC 25586 (F.n) and Porphyromonas gingivalis type strain ATCC 33277 (P.g 33277) bacterial species were tested when they were grown as monoculture or as dual species culture. (A) DNase I effect on biofilm formation, (B) DNase I effect on 48-h biofilm, (C) Proteinase K effect on biofilm formation, (D) Proteinase K effect on 48-h biofilm. The colored columns refer to the enzyme concentrations (0.125, 0.25, 0.5, and 1 mg/ml). The y-axis represents absorbance at 570 nm. The bars represent the means with standard deviations for 3–5 samples.

    Article Snippet: F. nucleatum type strain ATCC 25586 and P. gingivalis type strain ATCC 33277 were tested when they were grown as a monoculture or as a dual species culture.

    Techniques:

    Comparison of (A) carbohydrate and (B) eDNA yields from the biofilm matrix samples treated with proteinase K enzyme and non-treated samples. The matrix of 5-day-old biofilm was treated with 5 µg/ml proteinase K at 37°C for 1 h. F.n, Fusobacterium nucleatum ATCC 2558; P.g W50, Porphyromonas gingivalis W50; P.g 381, P. gingivalis FDC381; P.g 33277, P. gingivalis ATCC 33277. The bars represent the means with standard deviations from five samples (carbohydrates) and three replicates (eDNA).

    Journal: Journal of Oral Microbiology

    Article Title: Characterization of extracellular polymeric matrix, and treatment of Fusobacterium nucleatum and Porphyromonas gingivalis biofilms with DNase I and proteinase K

    doi: 10.3402/jom.v5i0.20015

    Figure Lengend Snippet: Comparison of (A) carbohydrate and (B) eDNA yields from the biofilm matrix samples treated with proteinase K enzyme and non-treated samples. The matrix of 5-day-old biofilm was treated with 5 µg/ml proteinase K at 37°C for 1 h. F.n, Fusobacterium nucleatum ATCC 2558; P.g W50, Porphyromonas gingivalis W50; P.g 381, P. gingivalis FDC381; P.g 33277, P. gingivalis ATCC 33277. The bars represent the means with standard deviations from five samples (carbohydrates) and three replicates (eDNA).

    Article Snippet: F. nucleatum type strain ATCC 25586 and P. gingivalis type strain ATCC 33277 were tested when they were grown as a monoculture or as a dual species culture.

    Techniques:

    Gel electrophoresis (0.8% agarose gel) of the extracellular DNA. The eDNA was extracted from the matrix of 5-day-old biofilm of these species. F.n, Fusobacterium nucleatum ATCC 25586; P.g W50, Porphyromonas gingivalis W50; P.g 381, P. gingivalis FDC381; P.g 33277, P. gingivalis ATCC 33277. Lane M, EZ load 100-bp molecular ruler (Bio-Rad).

    Journal: Journal of Oral Microbiology

    Article Title: Characterization of extracellular polymeric matrix, and treatment of Fusobacterium nucleatum and Porphyromonas gingivalis biofilms with DNase I and proteinase K

    doi: 10.3402/jom.v5i0.20015

    Figure Lengend Snippet: Gel electrophoresis (0.8% agarose gel) of the extracellular DNA. The eDNA was extracted from the matrix of 5-day-old biofilm of these species. F.n, Fusobacterium nucleatum ATCC 25586; P.g W50, Porphyromonas gingivalis W50; P.g 381, P. gingivalis FDC381; P.g 33277, P. gingivalis ATCC 33277. Lane M, EZ load 100-bp molecular ruler (Bio-Rad).

    Article Snippet: F. nucleatum type strain ATCC 25586 and P. gingivalis type strain ATCC 33277 were tested when they were grown as a monoculture or as a dual species culture.

    Techniques: Nucleic Acid Electrophoresis, Agarose Gel Electrophoresis

    Effect of DNase I and proteinase K enzymes at a concentration of 1 mg/ml on biofilm formation, (A) time zero and (B) 48-h-old biofilm. The graphs show the biomass (µm 3 /µm 2 ), maximum thickness (µm), and average thickness (µm) of one point z-stack confocal images analysed by COMSTAT software program. F.n, Fusobacterium nucleatum ATCC 25586; P.g 33277, Porphyromonas gingivalis ATCC 33277.

    Journal: Journal of Oral Microbiology

    Article Title: Characterization of extracellular polymeric matrix, and treatment of Fusobacterium nucleatum and Porphyromonas gingivalis biofilms with DNase I and proteinase K

    doi: 10.3402/jom.v5i0.20015

    Figure Lengend Snippet: Effect of DNase I and proteinase K enzymes at a concentration of 1 mg/ml on biofilm formation, (A) time zero and (B) 48-h-old biofilm. The graphs show the biomass (µm 3 /µm 2 ), maximum thickness (µm), and average thickness (µm) of one point z-stack confocal images analysed by COMSTAT software program. F.n, Fusobacterium nucleatum ATCC 25586; P.g 33277, Porphyromonas gingivalis ATCC 33277.

    Article Snippet: F. nucleatum type strain ATCC 25586 and P. gingivalis type strain ATCC 33277 were tested when they were grown as a monoculture or as a dual species culture.

    Techniques: Concentration Assay, Software

    Distribution of IS 1598 in several gram-negative bacteria. Southern hybridization of Pst I-digested genomic DNA from each strain was carried out with KIS-2 as a probe. Lanes: 1, P. gingivalis W83; 2, P. gingivalis ATCC 33277; 3, P. gingivalis W50; 4, P. gingivalis SU63; 5, P. gingivalis SUNY 1021; 6, P. gingivalis FDC 381; 7, P. gingivalis ESO 89; 8, P. asaccharolytica ATCC 25260; 9, P. loescheii ATCC 15930; 10, P. endodontalis ATCC 35406; 11, P. intermedia ATCC 25611; 12, C. ochracea S3; 13, F. nucleatum ATCC 25586; 14, A. actinomycetemcomitans Y4; 15, C. rectus ATCC 33238. P. gingivalis strains were classified based on AFNA and are indicated as “+” (AFNA positive) or “−” (AFNA negative) at the bottom of the panel.

    Journal: Infection and Immunity

    Article Title: Identification by Subtractive Hybridization of a Novel Insertion Sequence Specific for Virulent Strains of Porphyromonas gingivalis

    doi:

    Figure Lengend Snippet: Distribution of IS 1598 in several gram-negative bacteria. Southern hybridization of Pst I-digested genomic DNA from each strain was carried out with KIS-2 as a probe. Lanes: 1, P. gingivalis W83; 2, P. gingivalis ATCC 33277; 3, P. gingivalis W50; 4, P. gingivalis SU63; 5, P. gingivalis SUNY 1021; 6, P. gingivalis FDC 381; 7, P. gingivalis ESO 89; 8, P. asaccharolytica ATCC 25260; 9, P. loescheii ATCC 15930; 10, P. endodontalis ATCC 35406; 11, P. intermedia ATCC 25611; 12, C. ochracea S3; 13, F. nucleatum ATCC 25586; 14, A. actinomycetemcomitans Y4; 15, C. rectus ATCC 33238. P. gingivalis strains were classified based on AFNA and are indicated as “+” (AFNA positive) or “−” (AFNA negative) at the bottom of the panel.

    Article Snippet: The SH technique was used in this study, and a unique gene was detected in the virulent strain P. gingivalis W83 by comparing its chromosomal DNA with the DNA from the avirulent strain P. gingivalis ATCC 33277.

    Techniques: Hybridization

    Citrullinating activity in cell cultures. Activity of P. gingivalis strains containing variants PPAD‐T1 (ATCC 33277), PPAD‐T2 (ATCC T2; strain ATCC 33277 after introducing mutations G 231 N, E 232 T, and N 235 D), and a control, in which the antibiotic cassette used in ATCC T2 was introduced in wild‐type ATCC 33277 (ATCC T1 Ctrl). The results are shown as relative activity determined in triplicates using three independent cultures of each strain and displayed as mean ± SD. The statistical difference between strains was analyzed by the analysis of variance; ns, not significant; *** P

    Journal: Protein Science : A Publication of the Protein Society

    Article Title: Structure, function, and inhibition of a genomic/clinical variant of Porphyromonas gingivalis peptidylarginine deiminase

    doi: 10.1002/pro.3571

    Figure Lengend Snippet: Citrullinating activity in cell cultures. Activity of P. gingivalis strains containing variants PPAD‐T1 (ATCC 33277), PPAD‐T2 (ATCC T2; strain ATCC 33277 after introducing mutations G 231 N, E 232 T, and N 235 D), and a control, in which the antibiotic cassette used in ATCC T2 was introduced in wild‐type ATCC 33277 (ATCC T1 Ctrl). The results are shown as relative activity determined in triplicates using three independent cultures of each strain and displayed as mean ± SD. The statistical difference between strains was analyzed by the analysis of variance; ns, not significant; *** P

    Article Snippet: Along this line, the recent structure of PPAD from P. gingivalis reference strain ATCC 33277, hereafter PPAD‐T1, revealed the general architecture and modus operandi of this unique bacterial PAD., Moreover, recent studies reported that PPAD is present as point‐mutant variants differing from PPAD‐T1.

    Techniques: Activity Assay

    a A model of adding oral pathogen P. gingivalis proteins K2 and fluorescent dye (Alexa Fluor dextran 647, red colour ) to confluent gingival epithelial monolayers, to track pathological changes of epithelial integrity at a time course. Paracellular pathway of movement of the labelled dextran ( red ) was shown at 15 and 30 min observation times. b and c 2D z-stacking images in the beginning (T = 15 min) and end of the exposure time-lapse period (T = 1.5 h): left panels were the overlay images of cells ( green nuclei ) with far-red dye channel; right panels were far-red dye channel only. d 3D reconstruction time lapse images from ( b-c ) on live cells challenged with bacterial protein (K2) ( green colour : cell nuclei; red colour : dye). e Another example of projection of 2D z-stack imaging plus a position point. f A 3D reconstruction image with a position point ( e ) on live cells challenged with P. gingivalis bacterial protein (Ka) at 30 min ( green colour : Ka protein; red colour : nuclei)

    Journal: BMC Oral Health

    Article Title: Three/four-dimensional (3D/4D) microscopic imaging and processing in clinical dental research

    doi: 10.1186/s12903-016-0282-0

    Figure Lengend Snippet: a A model of adding oral pathogen P. gingivalis proteins K2 and fluorescent dye (Alexa Fluor dextran 647, red colour ) to confluent gingival epithelial monolayers, to track pathological changes of epithelial integrity at a time course. Paracellular pathway of movement of the labelled dextran ( red ) was shown at 15 and 30 min observation times. b and c 2D z-stacking images in the beginning (T = 15 min) and end of the exposure time-lapse period (T = 1.5 h): left panels were the overlay images of cells ( green nuclei ) with far-red dye channel; right panels were far-red dye channel only. d 3D reconstruction time lapse images from ( b-c ) on live cells challenged with bacterial protein (K2) ( green colour : cell nuclei; red colour : dye). e Another example of projection of 2D z-stack imaging plus a position point. f A 3D reconstruction image with a position point ( e ) on live cells challenged with P. gingivalis bacterial protein (Ka) at 30 min ( green colour : Ka protein; red colour : nuclei)

    Article Snippet: For drug development assay, oral pathogen P.gingivalis strain ATCC 33277 at MOI (multiplicity of infection) 100 cells per one epithelial cell [ ] was added to confluent H413 clone-1 epithelial monolayers for 1.5 h at 37 °C in 5 % CO2 .

    Techniques: Imaging

    Morphological changes observed in human gingival fibroblasts infected with P. gingivalis ATCC 33277 and gingipain-deficient mutants. (a) Control without bacteria; (b) ATCC 33277; (c) heat-treated ATCC 33277; (d) KDP112; (e) KDP128; (f) KDP129.

    Journal: Infection and Immunity

    Article Title: Effect of Inactivation of the Arg- and/or Lys-Gingipain Gene on Selected Virulence and Physiological Properties of Porphyromonas gingivalis

    doi: 10.1128/IAI.71.8.4742-4748.2003

    Figure Lengend Snippet: Morphological changes observed in human gingival fibroblasts infected with P. gingivalis ATCC 33277 and gingipain-deficient mutants. (a) Control without bacteria; (b) ATCC 33277; (c) heat-treated ATCC 33277; (d) KDP112; (e) KDP128; (f) KDP129.

    Article Snippet: This supports a recent study reporting that the growth rates of gingipain-deficient mutants and the parent P. gingivalis ATCC 33277 strain were similar when a chemically defined medium was supplemented with a protein hydrolysate instead of human serum albumin ( ).

    Techniques: Infection

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

    Journal: BMC Microbiology

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

    doi: 10.1186/1471-2180-13-206

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

    Article Snippet: Crude extract from P. gingivalis ATCC 33277 wild-type strain was obtained as previously described [ ] and prepared for use at a final concentration of 0.5 μg/mL.

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

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

    Journal: BMC Microbiology

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

    doi: 10.1186/1471-2180-13-206

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

    Article Snippet: Crude extract from P. gingivalis ATCC 33277 wild-type strain was obtained as previously described [ ] and prepared for use at a final concentration of 0.5 μg/mL.

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

    Maximum lesion size of mice challenged with either P. gingivalis ATCC 33277 (A) or W50 (B). BALB/c mice were immunized s.c. with the RgpA-Kgp proteinase complex (50 μg), formalin-killed (FK) P. gingivalis cells (2 × 10 8 of either strain ATCC 33277 or strain W50), formalin-killed E. coli cells (2 × 10 8 ), or PBS administered in IFA for both the primary and secondary doses. All mice were challenged 12 days after the secondary immunization with either P. gingivalis strain ATCC 33277 (7.5 × 10 9 viable cells) or W50 (3 × 10 9 viable cells) and were weighed, and the lesion sizes were measured daily over 14 days. Lesion sizes were statistically analyzed using Mann-Whitney U-Wilcoxon rank sum test. ∗, Groups significantly different ( P

    Journal: Infection and Immunity

    Article Title: RgpA-Kgp Peptide-Based Immunogens Provide Protection against Porphyromonas gingivalis Challenge in a Murine Lesion Model

    doi:

    Figure Lengend Snippet: Maximum lesion size of mice challenged with either P. gingivalis ATCC 33277 (A) or W50 (B). BALB/c mice were immunized s.c. with the RgpA-Kgp proteinase complex (50 μg), formalin-killed (FK) P. gingivalis cells (2 × 10 8 of either strain ATCC 33277 or strain W50), formalin-killed E. coli cells (2 × 10 8 ), or PBS administered in IFA for both the primary and secondary doses. All mice were challenged 12 days after the secondary immunization with either P. gingivalis strain ATCC 33277 (7.5 × 10 9 viable cells) or W50 (3 × 10 9 viable cells) and were weighed, and the lesion sizes were measured daily over 14 days. Lesion sizes were statistically analyzed using Mann-Whitney U-Wilcoxon rank sum test. ∗, Groups significantly different ( P

    Article Snippet: BALB/c mice were immunized with formalin-killed P. gingivalis strain ATCC 33277 or strain W50 (2 × 108 cells), formalin-killed E. coli cells (2 × 108 ), the RgpA-Kgp proteinase-adhesin complex (50 μg), or adjuvant alone (IFA).

    Techniques: Mouse Assay, Immunofluorescence, MANN-WHITNEY

    Binding to SB-Epoxy of MVs from different bacterial species. Shown are results of the assays using crude MV preparations from the following five different species: E. coli KP7600, N. meningitidis H44/76, P. aeruginosa PAO1 Holloway, C. perfringens strain 13, and P. gingivalis ATCC 33277. (A-D) Crude MVs from a single strain was incubated with the SB-Epoxy. Unbound components were collected in five washes and bound components were eventually eluted by SDS-PAGE loading buffer. Lanes denoted “1” are the starting material from conventional purification (Crude). Lanes denoted “2” are the first unbound fractions (Unbound, 1 st ). Lanes denoted “3” are the fifth unbound fractions (Unbound, 5 th ). Lanes denoted “4” are the elution fractions (Elu). (A: E. coli KP7600, B: N. meningitidis H44/76, C: P. aeruginosa PAO1-Tokai, D: C. perfringens strain 13). (E) Crude MVs from the five species were mixed with SB-Epoxy. The selective elimination of P. gingivalis MVs from the mixture is shown. Lanes 5 12: the input of mixtures of crude MVs (Crude mixture). Lanes 6 13: the first unbound fractions (Unbound, 1 st ). Lanes 12 13: five-fold diluted samples of lanes 5 6, respectively. Lanes 7, 8, 9: the third unbound fraction (Unbound, 3 rd ), the fifth unbound fraction (Unbound, 5 th ), and the elution fraction (Elu-Mix) from the mixture of four different MV preparations, respectively. Lane 10: the molecular weight marker. Lane 11: the crude MVs of P. gingivalis (Pg, Crude). Lane 14: the elution fraction from crude MVs of P. gingivalis only (Elu-Pg). The same volume (10 µl) was applied in each well of each PAGE gel. P. gingivalis FimA is denoted by an asterisk (*). N. meningitidis PorB is denoted by a dagger (†).

    Journal: PLoS ONE

    Article Title: A Novel Approach for Purification and Selective Capture of Membrane Vesicles of the Periodontopathic Bacterium, Porphyromonas gingivalis: Membrane Vesicles Bind to Magnetic Beads Coated with Epoxy Groups in a Noncovalent, Species-Specific Manner

    doi: 10.1371/journal.pone.0095137

    Figure Lengend Snippet: Binding to SB-Epoxy of MVs from different bacterial species. Shown are results of the assays using crude MV preparations from the following five different species: E. coli KP7600, N. meningitidis H44/76, P. aeruginosa PAO1 Holloway, C. perfringens strain 13, and P. gingivalis ATCC 33277. (A-D) Crude MVs from a single strain was incubated with the SB-Epoxy. Unbound components were collected in five washes and bound components were eventually eluted by SDS-PAGE loading buffer. Lanes denoted “1” are the starting material from conventional purification (Crude). Lanes denoted “2” are the first unbound fractions (Unbound, 1 st ). Lanes denoted “3” are the fifth unbound fractions (Unbound, 5 th ). Lanes denoted “4” are the elution fractions (Elu). (A: E. coli KP7600, B: N. meningitidis H44/76, C: P. aeruginosa PAO1-Tokai, D: C. perfringens strain 13). (E) Crude MVs from the five species were mixed with SB-Epoxy. The selective elimination of P. gingivalis MVs from the mixture is shown. Lanes 5 12: the input of mixtures of crude MVs (Crude mixture). Lanes 6 13: the first unbound fractions (Unbound, 1 st ). Lanes 12 13: five-fold diluted samples of lanes 5 6, respectively. Lanes 7, 8, 9: the third unbound fraction (Unbound, 3 rd ), the fifth unbound fraction (Unbound, 5 th ), and the elution fraction (Elu-Mix) from the mixture of four different MV preparations, respectively. Lane 10: the molecular weight marker. Lane 11: the crude MVs of P. gingivalis (Pg, Crude). Lane 14: the elution fraction from crude MVs of P. gingivalis only (Elu-Pg). The same volume (10 µl) was applied in each well of each PAGE gel. P. gingivalis FimA is denoted by an asterisk (*). N. meningitidis PorB is denoted by a dagger (†).

    Article Snippet: MV Preparation by the Conventional Method P. gingivalis strain ATCC 33277 was the primary strain used in this study, as many fimbriae are found in crude MV preparation.

    Techniques: Binding Assay, Incubation, SDS Page, Purification, Molecular Weight, Marker, Polyacrylamide Gel Electrophoresis

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

    Journal: BMC Microbiology

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

    doi: 10.1186/s12866-016-0810-8

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

    Article Snippet: Bacterial culture conditions P. gingivalis wild type strains ATCC 33277 (ATCC, Manassas, VA) and W50, and the W50-derived Kgp proteinase and Rgp proteinase mutant strains (K1A and E8, respectively) were a kind gift from Dr. M.A.

    Techniques: Binding Assay, SPR Assay, Incubation

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

    Journal: BMC Microbiology

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

    doi: 10.1186/s12866-016-0810-8

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

    Article Snippet: Bacterial culture conditions P. gingivalis wild type strains ATCC 33277 (ATCC, Manassas, VA) and W50, and the W50-derived Kgp proteinase and Rgp proteinase mutant strains (K1A and E8, respectively) were a kind gift from Dr. M.A.

    Techniques: Activity Assay

    Virulence of Porphyromonas gingivalis W83 and the sigP mutant strain in a murine model. Five BALB/c mice were inoculated subcutaneously with 0.1 mL of bacterial suspension at two sites on the depilated dorsal surface (0.2 mL per mouse), and the survival of the mice was monitored daily for up to 10 days. Three sets of experiments were carried out (15 mice in total). For the data analysis, Kaplan-Meier plots were constructed, and the log rank test was used to evaluate the differences in mean survival rates between mice infected with the W83 parent strain, sigP mutant, and complemented strain in three experiments.

    Journal: PLoS ONE

    Article Title: Effect of extracytoplasmic function sigma factors on autoaggregation, hemagglutination, and cell surface properties of Porphyromonas gingivalis

    doi: 10.1371/journal.pone.0185027

    Figure Lengend Snippet: Virulence of Porphyromonas gingivalis W83 and the sigP mutant strain in a murine model. Five BALB/c mice were inoculated subcutaneously with 0.1 mL of bacterial suspension at two sites on the depilated dorsal surface (0.2 mL per mouse), and the survival of the mice was monitored daily for up to 10 days. Three sets of experiments were carried out (15 mice in total). For the data analysis, Kaplan-Meier plots were constructed, and the log rank test was used to evaluate the differences in mean survival rates between mice infected with the W83 parent strain, sigP mutant, and complemented strain in three experiments.

    Article Snippet: For example, PG1318 in strain W83 [P . gingivalis ATCC 33277 open reading frame (ORF) number: PGN_1108] is involved in the regulation of mutation frequency in P . gingivalis [ ].

    Techniques: Mutagenesis, Mouse Assay, Construct, Infection

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

    Journal: The Scientific World Journal

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

    doi: 10.1155/2016/8626987

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

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

    Techniques: Infection

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

    Journal: Journal of Bacteriology

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

    doi: 10.1128/JB.00683-18

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

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

    Techniques: Mutagenesis

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

    Journal: Infection and Immunity

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

    doi: 10.1128/IAI.00648-09

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

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

    Techniques: Western Blot, Immunodetection, Incubation

    Citrullination of CXCL8 by different clinical isolates of P. gingivalis . Bacterial suspensions with starting concentrations of 5 × 10 7 CFU/ml were first grown for 24 h at 37°C in an anaerobic atmosphere. Then, the strains were treated

    Journal: Infection and Immunity

    Article Title: Citrullination and Proteolytic Processing of Chemokines by Porphyromonas gingivalis

    doi: 10.1128/IAI.01624-14

    Figure Lengend Snippet: Citrullination of CXCL8 by different clinical isolates of P. gingivalis . Bacterial suspensions with starting concentrations of 5 × 10 7 CFU/ml were first grown for 24 h at 37°C in an anaerobic atmosphere. Then, the strains were treated

    Article Snippet: P. gingivalis strains were grown on blood agar plates (Oxoid, Basingstoke, United Kingdom) supplemented with 5 μg/ml hemin (Sigma-Aldrich), 1 μg/ml menadione, and 5% sterile horse blood (Biotrading, Keerbergen, Belgium).

    Techniques:

    Levels of biotinylated CXCL8 measured in supernatants and cell pellets of P. gingivalis . Bacterial suspensions with starting concentrations of 5 × 10 7 CFU/ml were first grown for 24 h at 37°C in an anaerobic atmosphere. Then, the strains

    Journal: Infection and Immunity

    Article Title: Citrullination and Proteolytic Processing of Chemokines by Porphyromonas gingivalis

    doi: 10.1128/IAI.01624-14

    Figure Lengend Snippet: Levels of biotinylated CXCL8 measured in supernatants and cell pellets of P. gingivalis . Bacterial suspensions with starting concentrations of 5 × 10 7 CFU/ml were first grown for 24 h at 37°C in an anaerobic atmosphere. Then, the strains

    Article Snippet: P. gingivalis strains were grown on blood agar plates (Oxoid, Basingstoke, United Kingdom) supplemented with 5 μg/ml hemin (Sigma-Aldrich), 1 μg/ml menadione, and 5% sterile horse blood (Biotrading, Keerbergen, Belgium).

    Techniques:

    Recovery of CXCL8 after incubation with different clinical isolates and capsule-typed strains of P. gingivalis . Bacterial suspensions with starting concentrations of 5 × 10 7 CFU/ml were first grown for 24 h at 37°C in an anaerobic atmosphere.

    Journal: Infection and Immunity

    Article Title: Citrullination and Proteolytic Processing of Chemokines by Porphyromonas gingivalis

    doi: 10.1128/IAI.01624-14

    Figure Lengend Snippet: Recovery of CXCL8 after incubation with different clinical isolates and capsule-typed strains of P. gingivalis . Bacterial suspensions with starting concentrations of 5 × 10 7 CFU/ml were first grown for 24 h at 37°C in an anaerobic atmosphere.

    Article Snippet: P. gingivalis strains were grown on blood agar plates (Oxoid, Basingstoke, United Kingdom) supplemented with 5 μg/ml hemin (Sigma-Aldrich), 1 μg/ml menadione, and 5% sterile horse blood (Biotrading, Keerbergen, Belgium).

    Techniques: Incubation

    Proteolytic cleavage of chemokines by P. gingivalis .

    Journal: Infection and Immunity

    Article Title: Citrullination and Proteolytic Processing of Chemokines by Porphyromonas gingivalis

    doi: 10.1128/IAI.01624-14

    Figure Lengend Snippet: Proteolytic cleavage of chemokines by P. gingivalis .

    Article Snippet: P. gingivalis strains were grown on blood agar plates (Oxoid, Basingstoke, United Kingdom) supplemented with 5 μg/ml hemin (Sigma-Aldrich), 1 μg/ml menadione, and 5% sterile horse blood (Biotrading, Keerbergen, Belgium).

    Techniques:

    Cleavage of CXCL8 by the supernatants of different clinical isolates and capsule-typed strains of P. gingivalis . Bacterial suspensions with starting concentrations of 5 × 10 7 CFU/ml were grown for 24 h at 37°C in an anaerobic atmosphere,

    Journal: Infection and Immunity

    Article Title: Citrullination and Proteolytic Processing of Chemokines by Porphyromonas gingivalis

    doi: 10.1128/IAI.01624-14

    Figure Lengend Snippet: Cleavage of CXCL8 by the supernatants of different clinical isolates and capsule-typed strains of P. gingivalis . Bacterial suspensions with starting concentrations of 5 × 10 7 CFU/ml were grown for 24 h at 37°C in an anaerobic atmosphere,

    Article Snippet: P. gingivalis strains were grown on blood agar plates (Oxoid, Basingstoke, United Kingdom) supplemented with 5 μg/ml hemin (Sigma-Aldrich), 1 μg/ml menadione, and 5% sterile horse blood (Biotrading, Keerbergen, Belgium).

    Techniques:

    Circular map of the chromosome of P. gingivalis strain ATCC 33277. From the outside, the first and second circles show CDSs on the plus and minus strands, respectively. CDSs conserved in strains ATCC 33277 and W83 are indicated in red and ATCC 33277-specific CDSs in blue. The 3rd to 5th circles show IS elements (orange, IS Pg1 ; light green, IS Pg2 ; magenta, IS Pg3 ; cyan, IS Pg4 ; brown, IS Pg5 ; blue, IS Pg6 ), MITEs (magenta, MITE239; black, MITE PgRS ; cyan, MITE700), CTns, and Tns (blue, CTnPg1-a, CTnPg1-b, CTnPg2, and CTnPg3; red, Tn Pg17 ), respectively. The 6th and 7th circles show rrn operons and tRNA genes, respectively. The 8th circle shows the result of χ 2 analysis of nucleotide composition. Regions exhibiting values of > 600 are indicated in red and those of

    Journal: DNA Research: An International Journal for Rapid Publication of Reports on Genes and Genomes

    Article Title: Determination of the Genome Sequence of Porphyromonas gingivalis Strain ATCC 33277 and Genomic Comparison with Strain W83 Revealed Extensive Genome Rearrangements in P. gingivalis

    doi: 10.1093/dnares/dsn013

    Figure Lengend Snippet: Circular map of the chromosome of P. gingivalis strain ATCC 33277. From the outside, the first and second circles show CDSs on the plus and minus strands, respectively. CDSs conserved in strains ATCC 33277 and W83 are indicated in red and ATCC 33277-specific CDSs in blue. The 3rd to 5th circles show IS elements (orange, IS Pg1 ; light green, IS Pg2 ; magenta, IS Pg3 ; cyan, IS Pg4 ; brown, IS Pg5 ; blue, IS Pg6 ), MITEs (magenta, MITE239; black, MITE PgRS ; cyan, MITE700), CTns, and Tns (blue, CTnPg1-a, CTnPg1-b, CTnPg2, and CTnPg3; red, Tn Pg17 ), respectively. The 6th and 7th circles show rrn operons and tRNA genes, respectively. The 8th circle shows the result of χ 2 analysis of nucleotide composition. Regions exhibiting values of > 600 are indicated in red and those of

    Article Snippet: For preparing the genomic DNA, a single colony of each P. gingivalis strain was grown at 37°C anaerobically (10% CO2 , 10% H2 , 80% N2 ) in brain heart infusion broth (BD Bioscience, San Jose, CA, USA) supplemented with 5 µg of hemin and 0.5 µg of menadione per ml.

    Techniques:

    MITE in P. gingivalis with Repeating Structure (MITE PgRS ). ( A ) Schematic presentation of the consensus structure of MITE PgRS and the structures of 20 copies of MITE PgRS identified in the ATCC 33277 genome are shown. Three kind of repeat sequences, Repeats A, B, and C, are depicted by colored boxes. Red triangles indicate IR sequences and a black thick line in MITE PgRS _08 a unique nucleotide sequence. ( B ) Consensus sequences of Repeats A, B, and C are shown.

    Journal: DNA Research: An International Journal for Rapid Publication of Reports on Genes and Genomes

    Article Title: Determination of the Genome Sequence of Porphyromonas gingivalis Strain ATCC 33277 and Genomic Comparison with Strain W83 Revealed Extensive Genome Rearrangements in P. gingivalis

    doi: 10.1093/dnares/dsn013

    Figure Lengend Snippet: MITE in P. gingivalis with Repeating Structure (MITE PgRS ). ( A ) Schematic presentation of the consensus structure of MITE PgRS and the structures of 20 copies of MITE PgRS identified in the ATCC 33277 genome are shown. Three kind of repeat sequences, Repeats A, B, and C, are depicted by colored boxes. Red triangles indicate IR sequences and a black thick line in MITE PgRS _08 a unique nucleotide sequence. ( B ) Consensus sequences of Repeats A, B, and C are shown.

    Article Snippet: For preparing the genomic DNA, a single colony of each P. gingivalis strain was grown at 37°C anaerobically (10% CO2 , 10% H2 , 80% N2 ) in brain heart infusion broth (BD Bioscience, San Jose, CA, USA) supplemented with 5 µg of hemin and 0.5 µg of menadione per ml.

    Techniques: Sequencing

    Comparison of CRISPR-30-36 regions of P. gingivalis and B. fragilis . Locations and directions of CDSs (arrows) and repeat regions (black rectangles) are drawn to scale. Homologous CDSs are indicated by gray shading, and their amino-acid sequence identities are also shown. CDSs for IS transposases are indicated by black arrows, cas genes by vertically striped arrows, other functionally annotated CDSs by hatched arrows, and CDSs for hypothetical proteins by white arrows. The identity between PGN_1964 and PG2016 is 15.7%.

    Journal: DNA Research: An International Journal for Rapid Publication of Reports on Genes and Genomes

    Article Title: Determination of the Genome Sequence of Porphyromonas gingivalis Strain ATCC 33277 and Genomic Comparison with Strain W83 Revealed Extensive Genome Rearrangements in P. gingivalis

    doi: 10.1093/dnares/dsn013

    Figure Lengend Snippet: Comparison of CRISPR-30-36 regions of P. gingivalis and B. fragilis . Locations and directions of CDSs (arrows) and repeat regions (black rectangles) are drawn to scale. Homologous CDSs are indicated by gray shading, and their amino-acid sequence identities are also shown. CDSs for IS transposases are indicated by black arrows, cas genes by vertically striped arrows, other functionally annotated CDSs by hatched arrows, and CDSs for hypothetical proteins by white arrows. The identity between PGN_1964 and PG2016 is 15.7%.

    Article Snippet: For preparing the genomic DNA, a single colony of each P. gingivalis strain was grown at 37°C anaerobically (10% CO2 , 10% H2 , 80% N2 ) in brain heart infusion broth (BD Bioscience, San Jose, CA, USA) supplemented with 5 µg of hemin and 0.5 µg of menadione per ml.

    Techniques: CRISPR, Sequencing

    The DNA sequence identity plot of P. gingivalis ATCC 33277 and W83 chromosomes. The dnaA gene is located at the left and bottom corner. Black circles indicate mobile genetic elements (CTn Tn, IS, MITE, or a not-well-defined large mobile element of W83). The chromosomal locations of other genetic elements that mediated inversions or translocations are shown in the right: rrn operons (black squares), duplicated regions coding for a histone-like DNA binding protein, a hypothetical protein and elongation factor P (open squares), 12/13 bp repeat sequences (black triangle), and 11 bp repeat sequences (open triangle).

    Journal: DNA Research: An International Journal for Rapid Publication of Reports on Genes and Genomes

    Article Title: Determination of the Genome Sequence of Porphyromonas gingivalis Strain ATCC 33277 and Genomic Comparison with Strain W83 Revealed Extensive Genome Rearrangements in P. gingivalis

    doi: 10.1093/dnares/dsn013

    Figure Lengend Snippet: The DNA sequence identity plot of P. gingivalis ATCC 33277 and W83 chromosomes. The dnaA gene is located at the left and bottom corner. Black circles indicate mobile genetic elements (CTn Tn, IS, MITE, or a not-well-defined large mobile element of W83). The chromosomal locations of other genetic elements that mediated inversions or translocations are shown in the right: rrn operons (black squares), duplicated regions coding for a histone-like DNA binding protein, a hypothetical protein and elongation factor P (open squares), 12/13 bp repeat sequences (black triangle), and 11 bp repeat sequences (open triangle).

    Article Snippet: For preparing the genomic DNA, a single colony of each P. gingivalis strain was grown at 37°C anaerobically (10% CO2 , 10% H2 , 80% N2 ) in brain heart infusion broth (BD Bioscience, San Jose, CA, USA) supplemented with 5 µg of hemin and 0.5 µg of menadione per ml.

    Techniques: Sequencing, Binding Assay

    Inactivation of PG1828 in P. gingivalis . (A) Construction of a PG1828-deficient mutant of P. gingivalis strain 381 by allelic exchange mutagenesis with an ermF-ermAM antibiotic resistance cassette (as described in Materials and Methods). (B) Predicted maps of the genomes of the wild-type and the PG1828-deficient mutant. Arrowheads indicate the numbers and positions of oligonucleotide primers for PCR analysis (as described in Materials and Methods). (C) PCR analysis of WT and ΔPG1828. Numbers above the lanes indicate the primer pairs (B) used for the PCR analysis. Lane M, DNA marker; left margin, molecular sizes.

    Journal: Infection and Immunity

    Article Title: Lipopolysaccharide Preparation Extracted from Porphyromonas gingivalis Lipoprotein-Deficient Mutant Shows a Marked Decrease in Toll-Like Receptor 2-Mediated Signaling

    doi: 10.1128/IAI.73.4.2157-2163.2005

    Figure Lengend Snippet: Inactivation of PG1828 in P. gingivalis . (A) Construction of a PG1828-deficient mutant of P. gingivalis strain 381 by allelic exchange mutagenesis with an ermF-ermAM antibiotic resistance cassette (as described in Materials and Methods). (B) Predicted maps of the genomes of the wild-type and the PG1828-deficient mutant. Arrowheads indicate the numbers and positions of oligonucleotide primers for PCR analysis (as described in Materials and Methods). (C) PCR analysis of WT and ΔPG1828. Numbers above the lanes indicate the primer pairs (B) used for the PCR analysis. Lane M, DNA marker; left margin, molecular sizes.

    Article Snippet: P. gingivalis strain 381 was grown anaerobically at 37°C in brain heart infusion (BHI) broth (Difco, Detroit, Mich.) containing 0.5% yeast extract (Difco), 5 μg of hemin per ml, and 1 μg of vitamin K3 per ml.

    Techniques: Mutagenesis, Polymerase Chain Reaction, Marker

    Representative microscopic images of P. gingivalis strain W83 within LC3 positive or LAMP-1 positive vacuoles at 6 hours post-inoculation. Arrows indicate magnified inserts. Scale bar is equivalent to 10 µm. Representative images of other P. gingivalis strains can be viewed in supporting file Figure S4 .

    Journal: PLoS ONE

    Article Title: Porphyromonas gingivalis Strain Specific Interactions with Human Coronary Artery Endothelial Cells: A Comparative Study

    doi: 10.1371/journal.pone.0052606

    Figure Lengend Snippet: Representative microscopic images of P. gingivalis strain W83 within LC3 positive or LAMP-1 positive vacuoles at 6 hours post-inoculation. Arrows indicate magnified inserts. Scale bar is equivalent to 10 µm. Representative images of other P. gingivalis strains can be viewed in supporting file Figure S4 .

    Article Snippet: P. gingivalis was detected with a rabbit polyclonal antibody that was produced by immunization with whole cells of P. gingivalis strain W83 [Lot number C7947 – produced by Strategic Biosolutions, Newark, DE].

    Techniques:

    Adherence of P. gingivalis to HCAE cells detected by qPCR (A) and ELISA (B). Values represent the mean ± SD of six biological replicates from two independent experiments. Inoculated HCAE cells were incubated at 4°C for 30 min without agitation. (A) P. gingivalis 16S DNA copy number was normalized to HCAE cell 18S copy number. (B) Absorbance values at 450 nm for each replicate were normalized to their corresponding inoculums. **Values were significantly less than W83 and 381 ( P

    Journal: PLoS ONE

    Article Title: Porphyromonas gingivalis Strain Specific Interactions with Human Coronary Artery Endothelial Cells: A Comparative Study

    doi: 10.1371/journal.pone.0052606

    Figure Lengend Snippet: Adherence of P. gingivalis to HCAE cells detected by qPCR (A) and ELISA (B). Values represent the mean ± SD of six biological replicates from two independent experiments. Inoculated HCAE cells were incubated at 4°C for 30 min without agitation. (A) P. gingivalis 16S DNA copy number was normalized to HCAE cell 18S copy number. (B) Absorbance values at 450 nm for each replicate were normalized to their corresponding inoculums. **Values were significantly less than W83 and 381 ( P

    Article Snippet: P. gingivalis was detected with a rabbit polyclonal antibody that was produced by immunization with whole cells of P. gingivalis strain W83 [Lot number C7947 – produced by Strategic Biosolutions, Newark, DE].

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

    Effect of Lys 129 mutation on Kgp expression and activity. Western blotting analysis of parental strain P. gingivalis W83 and mutants K129A, K129E, and K129R is shown. Late exponential/early stationary bacterial cultures (BC) were separated by centrifugation into cell-free growth medium (GM) and cell pellet. The latter was washed, giving rise to the whole cell (WC) fraction, which was further fractionated into the soluble intracellular protein fraction (CP + PP) and the cell envelope fracion (CE) by sonication and ultracentrifugation. A and B , all fractions were standardized to the initial volume of the culture subjected to centrifugation and analyzed by Western blotting to detect Kgp forms ( A ) and Rgps (control) ( B ). C and D , Kgp ( C ) and Rgp ( D ) gingipain activities determined in whole cultures, cell-free growth medium, and washed cells using specific substrates. The whole-culture activity of the wild-type strain was arbitrarily taken as 100%.

    Journal: The Journal of Biological Chemistry

    Article Title: Structural insights unravel the zymogenic mechanism of the virulence factor gingipain K from Porphyromonas gingivalis, a causative agent of gum disease from the human oral microbiome *

    doi: 10.1074/jbc.M117.776724

    Figure Lengend Snippet: Effect of Lys 129 mutation on Kgp expression and activity. Western blotting analysis of parental strain P. gingivalis W83 and mutants K129A, K129E, and K129R is shown. Late exponential/early stationary bacterial cultures (BC) were separated by centrifugation into cell-free growth medium (GM) and cell pellet. The latter was washed, giving rise to the whole cell (WC) fraction, which was further fractionated into the soluble intracellular protein fraction (CP + PP) and the cell envelope fracion (CE) by sonication and ultracentrifugation. A and B , all fractions were standardized to the initial volume of the culture subjected to centrifugation and analyzed by Western blotting to detect Kgp forms ( A ) and Rgps (control) ( B ). C and D , Kgp ( C ) and Rgp ( D ) gingipain activities determined in whole cultures, cell-free growth medium, and washed cells using specific substrates. The whole-culture activity of the wild-type strain was arbitrarily taken as 100%.

    Article Snippet: Briefly, 1 μg of purified plasmid DNA was electroporated into P. gingivalis strain W83 competent cells (2.5 kV, 4 ms; Bio-Rad Micropulser).

    Techniques: Mutagenesis, Expressing, Activity Assay, Western Blot, Centrifugation, Sonication

    P. gingivalis enhances IL-1β secretion and caspase-1 activation, leading to inflammatory cell death. (A) PMA-primed THP-1 cells were infected with P. gingivalis (MOI, 100) for 6 or 24 h. Cell culture supernatants were assayed for human IL-1β

    Journal: Infection and Immunity

    Article Title: Activation of NLRP3 and AIM2 Inflammasomes by Porphyromonas gingivalis Infection

    doi: 10.1128/IAI.00862-13

    Figure Lengend Snippet: P. gingivalis enhances IL-1β secretion and caspase-1 activation, leading to inflammatory cell death. (A) PMA-primed THP-1 cells were infected with P. gingivalis (MOI, 100) for 6 or 24 h. Cell culture supernatants were assayed for human IL-1β

    Article Snippet: P. gingivalis (strain 381) was grown in Gifu anaerobic medium broth (Nissui, Japan), which contained 5 mg/ml hemin and 0.5 mg/ml 3-phytyl-menadione (vitamin K) under anaerobic conditions at 37°C.

    Techniques: Activation Assay, Infection, Cell Culture

    P. gingivalis -induced IL-1β secretion and proinflammatory cell death are dependent on caspase-1. (A to C) THP-1 cells were pretreated with Z-WEHD-FMK (30 μM) or Z-VAD-FMK (30 μM) for 30 min before P. gingivalis infection for 6

    Journal: Infection and Immunity

    Article Title: Activation of NLRP3 and AIM2 Inflammasomes by Porphyromonas gingivalis Infection

    doi: 10.1128/IAI.00862-13

    Figure Lengend Snippet: P. gingivalis -induced IL-1β secretion and proinflammatory cell death are dependent on caspase-1. (A to C) THP-1 cells were pretreated with Z-WEHD-FMK (30 μM) or Z-VAD-FMK (30 μM) for 30 min before P. gingivalis infection for 6

    Article Snippet: P. gingivalis (strain 381) was grown in Gifu anaerobic medium broth (Nissui, Japan), which contained 5 mg/ml hemin and 0.5 mg/ml 3-phytyl-menadione (vitamin K) under anaerobic conditions at 37°C.

    Techniques: Infection

    P. gingivalis -induced inflammasome activation requires the preceding TLR signaling. (A to C) THP-1 cells were transfected with TLR2 and/or TLR4 siRNA for 48 h. siRNA-transfected cells were infected with P. gingivalis (MOI, 100) for 6 h. The IL-1β

    Journal: Infection and Immunity

    Article Title: Activation of NLRP3 and AIM2 Inflammasomes by Porphyromonas gingivalis Infection

    doi: 10.1128/IAI.00862-13

    Figure Lengend Snippet: P. gingivalis -induced inflammasome activation requires the preceding TLR signaling. (A to C) THP-1 cells were transfected with TLR2 and/or TLR4 siRNA for 48 h. siRNA-transfected cells were infected with P. gingivalis (MOI, 100) for 6 h. The IL-1β

    Article Snippet: P. gingivalis (strain 381) was grown in Gifu anaerobic medium broth (Nissui, Japan), which contained 5 mg/ml hemin and 0.5 mg/ml 3-phytyl-menadione (vitamin K) under anaerobic conditions at 37°C.

    Techniques: Activation Assay, Transfection, Infection

    Caspase-1 activation, IL-1β secretion, cell cytotoxicity, and pyroptic cell death induced by P. gingivalis infection are mediated by NLRP3 and AIM2 inflammasome activation. (A) Real-time PCR measurement of ASC, NLRP3, AIM2, NLRPC4, and IL-1β

    Journal: Infection and Immunity

    Article Title: Activation of NLRP3 and AIM2 Inflammasomes by Porphyromonas gingivalis Infection

    doi: 10.1128/IAI.00862-13

    Figure Lengend Snippet: Caspase-1 activation, IL-1β secretion, cell cytotoxicity, and pyroptic cell death induced by P. gingivalis infection are mediated by NLRP3 and AIM2 inflammasome activation. (A) Real-time PCR measurement of ASC, NLRP3, AIM2, NLRPC4, and IL-1β

    Article Snippet: P. gingivalis (strain 381) was grown in Gifu anaerobic medium broth (Nissui, Japan), which contained 5 mg/ml hemin and 0.5 mg/ml 3-phytyl-menadione (vitamin K) under anaerobic conditions at 37°C.

    Techniques: Activation Assay, Infection, Real-time Polymerase Chain Reaction

    Infection with P. gingivalis triggers the activation of NLRP3 and AIM2 inflammasomes via TLR2 and TLR4 signaling, leading to IL-1β secretion and pyroptic cell death.

    Journal: Infection and Immunity

    Article Title: Activation of NLRP3 and AIM2 Inflammasomes by Porphyromonas gingivalis Infection

    doi: 10.1128/IAI.00862-13

    Figure Lengend Snippet: Infection with P. gingivalis triggers the activation of NLRP3 and AIM2 inflammasomes via TLR2 and TLR4 signaling, leading to IL-1β secretion and pyroptic cell death.

    Article Snippet: P. gingivalis (strain 381) was grown in Gifu anaerobic medium broth (Nissui, Japan), which contained 5 mg/ml hemin and 0.5 mg/ml 3-phytyl-menadione (vitamin K) under anaerobic conditions at 37°C.

    Techniques: Infection, Activation Assay

    ATP and potassium (K + ) efflux are involved in P. gingivalis -induced NLRP3 activation. (A) THP-1 cells were infected with P. gingivalis for 6 h. The extracellular ATP concentration was determined using an ATP determination kit. The data are shown as mean

    Journal: Infection and Immunity

    Article Title: Activation of NLRP3 and AIM2 Inflammasomes by Porphyromonas gingivalis Infection

    doi: 10.1128/IAI.00862-13

    Figure Lengend Snippet: ATP and potassium (K + ) efflux are involved in P. gingivalis -induced NLRP3 activation. (A) THP-1 cells were infected with P. gingivalis for 6 h. The extracellular ATP concentration was determined using an ATP determination kit. The data are shown as mean

    Article Snippet: P. gingivalis (strain 381) was grown in Gifu anaerobic medium broth (Nissui, Japan), which contained 5 mg/ml hemin and 0.5 mg/ml 3-phytyl-menadione (vitamin K) under anaerobic conditions at 37°C.

    Techniques: Activation Assay, Infection, Concentration Assay

    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: Individuals are most often colonized by one strain of P. gingivalis , although it is possible to be transiently coinfected by two or three strains simultaneously ( , ).

    Techniques:

    Electron micrograph showing changes in surface structures of P. gingivalis ATCC 33277 and W83. Bacterial cells grown to the log phase (OD 600 of 0.7–0.9) were processed for electron microscopic examination using formvar-carbon coated grids (500 mesh) and were examined using Philips Tecnai 12 TEM. Fimbriae were lacking in the vimF mutant FLL476 when compared with the wild ATCC33277 and the complemented strain FLL476C’. A thick glycocalyx along with vesicles and a well-defined outer membrane was observed in W83. FLL95 showed hazy outer membrane with reduced visicles. In the complemented strain FLL95C’ the outer membrane morphology was restored.

    Journal: PLoS ONE

    Article Title: In Porphyromonas gingivalis VimF Is Involved in Gingipain Maturation through the Transfer of Galactose

    doi: 10.1371/journal.pone.0063367

    Figure Lengend Snippet: Electron micrograph showing changes in surface structures of P. gingivalis ATCC 33277 and W83. Bacterial cells grown to the log phase (OD 600 of 0.7–0.9) were processed for electron microscopic examination using formvar-carbon coated grids (500 mesh) and were examined using Philips Tecnai 12 TEM. Fimbriae were lacking in the vimF mutant FLL476 when compared with the wild ATCC33277 and the complemented strain FLL476C’. A thick glycocalyx along with vesicles and a well-defined outer membrane was observed in W83. FLL95 showed hazy outer membrane with reduced visicles. In the complemented strain FLL95C’ the outer membrane morphology was restored.

    Article Snippet: Bacterial Growth Conditions and Gingipain Assays All strains of P. gingivalis were grown in brain heart infusion (BHI) broth (Difco Laboratories, Detroit, MI) supplemented with hemin (5 µg/ml), vitamin K (0.5 µg/ml) and cysteine (0.1%).

    Techniques: Transmission Electron Microscopy, Mutagenesis

    Comparison of biofilm formation, autoaggregation, hemagglutination and invasion assay. A. Biofilm formation of ATCC 33277, FLL476 and FLL476C’ were compared. Biofilm assay was done by staining adherent cells of overnight cultures grown in microtiter plates with 0.5% (w/v) crystal violet. Blank contained only media. Biofilm forming ability corresponded to OD 595 . B. Autoaggregation of 33277, FLL476 and FLL476C’ corresponded to change in OD 600 monitored for about three hours after cells were washed and suspended in PBS. A representative sample is shown. C. Hemagglutination activities of ATCC 33277, FLL476 and FLL476C’ were assessed by serially diluting cells in PBS and incubating with sheep RBCs for 3 h at 4°C. Dilutions are listed above and last dilution showing matt formation was taken as the titer. The blank contained only media. D. Antibiotic Protection Assay was used to quantify invasion. P. gingivalis cells that were able to invade HeLa cell monolayers were released by lysis and cultured on BA plates. Infectivity was taken as the percentage of cells recovered. (* = p

    Journal: PLoS ONE

    Article Title: In Porphyromonas gingivalis VimF Is Involved in Gingipain Maturation through the Transfer of Galactose

    doi: 10.1371/journal.pone.0063367

    Figure Lengend Snippet: Comparison of biofilm formation, autoaggregation, hemagglutination and invasion assay. A. Biofilm formation of ATCC 33277, FLL476 and FLL476C’ were compared. Biofilm assay was done by staining adherent cells of overnight cultures grown in microtiter plates with 0.5% (w/v) crystal violet. Blank contained only media. Biofilm forming ability corresponded to OD 595 . B. Autoaggregation of 33277, FLL476 and FLL476C’ corresponded to change in OD 600 monitored for about three hours after cells were washed and suspended in PBS. A representative sample is shown. C. Hemagglutination activities of ATCC 33277, FLL476 and FLL476C’ were assessed by serially diluting cells in PBS and incubating with sheep RBCs for 3 h at 4°C. Dilutions are listed above and last dilution showing matt formation was taken as the titer. The blank contained only media. D. Antibiotic Protection Assay was used to quantify invasion. P. gingivalis cells that were able to invade HeLa cell monolayers were released by lysis and cultured on BA plates. Infectivity was taken as the percentage of cells recovered. (* = p

    Article Snippet: Bacterial Growth Conditions and Gingipain Assays All strains of P. gingivalis were grown in brain heart infusion (BHI) broth (Difco Laboratories, Detroit, MI) supplemented with hemin (5 µg/ml), vitamin K (0.5 µg/ml) and cysteine (0.1%).

    Techniques: Invasion Assay, Biofilm Production Assay, Staining, Lysis, Cell Culture, Infection

    Comparison of growth and gingipain activities of wild-type, vimF mutant and complemented strains of W83 and ATCC 33277. Growth rate of P. gingivalis ATCC 33277 ( A ) and W83 ( C ) were compared with their respective vimF -defective isogenic mutants (FLL476 and FLL95 ) and complemented strains (FLL476C’ and FLL95C’). The data shown is an average of three independent replicates. Error bars represent the SD. Gingipain activity of W83 ( D ) and ATCC 33277 ( B ) were compared with respective mutants and complemented strains. The activities were normalized to W83 and ATCC 33277 being 100% and the mutants reported as a percentage thereof. Error bars represent SD.

    Journal: PLoS ONE

    Article Title: In Porphyromonas gingivalis VimF Is Involved in Gingipain Maturation through the Transfer of Galactose

    doi: 10.1371/journal.pone.0063367

    Figure Lengend Snippet: Comparison of growth and gingipain activities of wild-type, vimF mutant and complemented strains of W83 and ATCC 33277. Growth rate of P. gingivalis ATCC 33277 ( A ) and W83 ( C ) were compared with their respective vimF -defective isogenic mutants (FLL476 and FLL95 ) and complemented strains (FLL476C’ and FLL95C’). The data shown is an average of three independent replicates. Error bars represent the SD. Gingipain activity of W83 ( D ) and ATCC 33277 ( B ) were compared with respective mutants and complemented strains. The activities were normalized to W83 and ATCC 33277 being 100% and the mutants reported as a percentage thereof. Error bars represent SD.

    Article Snippet: Bacterial Growth Conditions and Gingipain Assays All strains of P. gingivalis were grown in brain heart infusion (BHI) broth (Difco Laboratories, Detroit, MI) supplemented with hemin (5 µg/ml), vitamin K (0.5 µg/ml) and cysteine (0.1%).

    Techniques: Mutagenesis, Activity Assay

    Proliferation and differentiation of CD4 + T cells mediated by gingipains from P. gingivalis . Monocyte-derived dendritic cells (moDC) were untreated or exposed to P. gingivalis W83 in the presence/absence of specific protease inhibitors (KYT-1 and KYT-36, each at a concentration of 1 μM) or the isogenic gingipain-null mutant ΔKΔRAB for 6 h at MOI (multiplicity of infection) 1:50. Naive CD4 + T cells stained with CFSE were added and co-stimulated for 5 days. Proliferation of lymphocytes was measured at day 1, 3, and 5. (A) Completed results of proliferation on day 1, 3, and 5 as a percentage of CFSE low cells. Data are presented as a mean ± standard deviation of assays performed in triplicate using four independent donors, and were analyzed with a two way ANOVA with the Bonferoni's posttest correction (# P

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Inactive Gingipains from P. gingivalis Selectively Skews T Cells toward a Th17 Phenotype in an IL-6 Dependent Manner

    doi: 10.3389/fcimb.2017.00140

    Figure Lengend Snippet: Proliferation and differentiation of CD4 + T cells mediated by gingipains from P. gingivalis . Monocyte-derived dendritic cells (moDC) were untreated or exposed to P. gingivalis W83 in the presence/absence of specific protease inhibitors (KYT-1 and KYT-36, each at a concentration of 1 μM) or the isogenic gingipain-null mutant ΔKΔRAB for 6 h at MOI (multiplicity of infection) 1:50. Naive CD4 + T cells stained with CFSE were added and co-stimulated for 5 days. Proliferation of lymphocytes was measured at day 1, 3, and 5. (A) Completed results of proliferation on day 1, 3, and 5 as a percentage of CFSE low cells. Data are presented as a mean ± standard deviation of assays performed in triplicate using four independent donors, and were analyzed with a two way ANOVA with the Bonferoni's posttest correction (# P

    Article Snippet: All characterized strains of P. gingivalis , as well as clinical isolates, produce gingipains (Ismail et al., ), which are major virulence factors capable of manipulation of a large number of defense and homeostatic mechanisms in infected tissue.

    Techniques: Derivative Assay, Concentration Assay, Mutagenesis, Infection, Staining, Standard Deviation

    Activation of Th17 signaling pathway in CD4+ naïve lymphocyte depends on gingipain activity . Monocyte-derived dendritic cells (moDC) were untreated or exposed to P. gingivalis in the presence or absence of specific protease inhibitors (KYT-1 and KYT-36, each at a concentration of 1 μM). After 6 h, CD4+ naïve lymphocytes were added and co-stimulated for 3 consecutive days. At day 1, 2, and 3 after co-incubation, cells were collected and lysed with TRIzol, RNA was isolated and reverse transcriptase PCR was performed. Relative expression of cytokine genes IL-6R, IL-1R, TGF β R, RORc, ROR α , STAT3, RUNX1, IRF4, BATF to the reference house-keeping gene EF2 was measured by Real-Time PCR. Data represent fold increase in expression compared to control levels, which were arbitrarily set at 1 and were analyzed with a Student's t -test ( # P

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Inactive Gingipains from P. gingivalis Selectively Skews T Cells toward a Th17 Phenotype in an IL-6 Dependent Manner

    doi: 10.3389/fcimb.2017.00140

    Figure Lengend Snippet: Activation of Th17 signaling pathway in CD4+ naïve lymphocyte depends on gingipain activity . Monocyte-derived dendritic cells (moDC) were untreated or exposed to P. gingivalis in the presence or absence of specific protease inhibitors (KYT-1 and KYT-36, each at a concentration of 1 μM). After 6 h, CD4+ naïve lymphocytes were added and co-stimulated for 3 consecutive days. At day 1, 2, and 3 after co-incubation, cells were collected and lysed with TRIzol, RNA was isolated and reverse transcriptase PCR was performed. Relative expression of cytokine genes IL-6R, IL-1R, TGF β R, RORc, ROR α , STAT3, RUNX1, IRF4, BATF to the reference house-keeping gene EF2 was measured by Real-Time PCR. Data represent fold increase in expression compared to control levels, which were arbitrarily set at 1 and were analyzed with a Student's t -test ( # P

    Article Snippet: All characterized strains of P. gingivalis , as well as clinical isolates, produce gingipains (Ismail et al., ), which are major virulence factors capable of manipulation of a large number of defense and homeostatic mechanisms in infected tissue.

    Techniques: Activation Assay, Activity Assay, Derivative Assay, Concentration Assay, Incubation, Isolation, Polymerase Chain Reaction, Expressing, Real-time Polymerase Chain Reaction

    Blocking the IL-6 signaling pathway results in reduction of the Th17 population stimulated by gingipains . Monocyte-derived dendritic cells were untreated or exposed to P. gingivalis W83 in the presence/absence of specific protease inhibitors (KYT-1 and KYT-36, each at a concentration of 1 μM) for 6 h. Cultures were treated with the inhibitor of JAK1/JAK2 (Ruxolitinib [1 μM]) (A) or antibodies against the IL-6 receptor [10 μg/ml]) (B) for 2 h then naive CD4 + cells were added and cells were co-cultured for 5 or 7 days. At the end of each period, T cells were stimulated with PMA and ionomycin for 4 h in the presence of GolgiStop. For the evaluation of the Th17 population, cells were stained for IL-17A and RORγt, and analyzed by flow cytometry. Data show transcription factor and cytokine positive cells (RORγt + IL-17A + ) and are presented as a mean ± standard deviation of assays performed using three independent donors. Data were analyzed with a Student's t -test ( * P

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Inactive Gingipains from P. gingivalis Selectively Skews T Cells toward a Th17 Phenotype in an IL-6 Dependent Manner

    doi: 10.3389/fcimb.2017.00140

    Figure Lengend Snippet: Blocking the IL-6 signaling pathway results in reduction of the Th17 population stimulated by gingipains . Monocyte-derived dendritic cells were untreated or exposed to P. gingivalis W83 in the presence/absence of specific protease inhibitors (KYT-1 and KYT-36, each at a concentration of 1 μM) for 6 h. Cultures were treated with the inhibitor of JAK1/JAK2 (Ruxolitinib [1 μM]) (A) or antibodies against the IL-6 receptor [10 μg/ml]) (B) for 2 h then naive CD4 + cells were added and cells were co-cultured for 5 or 7 days. At the end of each period, T cells were stimulated with PMA and ionomycin for 4 h in the presence of GolgiStop. For the evaluation of the Th17 population, cells were stained for IL-17A and RORγt, and analyzed by flow cytometry. Data show transcription factor and cytokine positive cells (RORγt + IL-17A + ) and are presented as a mean ± standard deviation of assays performed using three independent donors. Data were analyzed with a Student's t -test ( * P

    Article Snippet: All characterized strains of P. gingivalis , as well as clinical isolates, produce gingipains (Ismail et al., ), which are major virulence factors capable of manipulation of a large number of defense and homeostatic mechanisms in infected tissue.

    Techniques: Blocking Assay, Derivative Assay, Concentration Assay, Cell Culture, Staining, Flow Cytometry, Cytometry, Standard Deviation

    Interleukin 6 production induced by P. gingivalis is inversely dependent on gingipain activity . Monocyte-derived dendritic cells (moDC) were untreated or exposed to P. gingivalis W83 in the presence or absence of specific protease inhibitors (KYT-1 and KYT-36, each at 1 μM concentration) or isogenic P. gingivalis gingipain-null mutant ΔKΔRAB for 2 h (A,D,G) , 6 h (B,E,H) , and 24 h ( C , F , I ). (A–C) Supernatants were collected and the concentration of IL-6 was determined by ELISA. (D–F) Arginine-specific and (G–I) Lysine-specific gingipain activity was determined using L-BA p Na and N-( p -Tosyl)-Gly-Pro-Lys-4-nitroanilide acetate salt (200 μM) as a substrate, respectively. Data are presented as means ± standard deviations of assays performed in triplicate using three independent donors, and were analyzed by one-way ANOVA with the Bonferroni post-test correction ( # P

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Inactive Gingipains from P. gingivalis Selectively Skews T Cells toward a Th17 Phenotype in an IL-6 Dependent Manner

    doi: 10.3389/fcimb.2017.00140

    Figure Lengend Snippet: Interleukin 6 production induced by P. gingivalis is inversely dependent on gingipain activity . Monocyte-derived dendritic cells (moDC) were untreated or exposed to P. gingivalis W83 in the presence or absence of specific protease inhibitors (KYT-1 and KYT-36, each at 1 μM concentration) or isogenic P. gingivalis gingipain-null mutant ΔKΔRAB for 2 h (A,D,G) , 6 h (B,E,H) , and 24 h ( C , F , I ). (A–C) Supernatants were collected and the concentration of IL-6 was determined by ELISA. (D–F) Arginine-specific and (G–I) Lysine-specific gingipain activity was determined using L-BA p Na and N-( p -Tosyl)-Gly-Pro-Lys-4-nitroanilide acetate salt (200 μM) as a substrate, respectively. Data are presented as means ± standard deviations of assays performed in triplicate using three independent donors, and were analyzed by one-way ANOVA with the Bonferroni post-test correction ( # P

    Article Snippet: All characterized strains of P. gingivalis , as well as clinical isolates, produce gingipains (Ismail et al., ), which are major virulence factors capable of manipulation of a large number of defense and homeostatic mechanisms in infected tissue.

    Techniques: Activity Assay, Derivative Assay, Concentration Assay, Mutagenesis, Enzyme-linked Immunosorbent Assay

    The role of gingipains in the regulation of cell responses to LPS and fimbriae . Monocyte-derived dendritic cells (moDC) were untreated or exposed to P. gingivalis LPS (2.5 μg/ml), FimA (10 μg/ml) or mixed LPS with FimA. Additionally, cells were co-stimulated with HRgpA (2 nM) in the presence or absence of specific protease inhibitors (KYT-1 and KYT-36, each at a concentration of 1 μM). Four hours after stimulation, culture media were collected and cells were lysed with TRIzol. RNA was isolated and reverse transcriptase PCR was performed. Relative expression of the cytokine gene IL-6 to the reference house-keeping gene EF2 was measured by Real-Time PCR. Data show representative fold increase in expression compared to control levels, which were arbitrarily set at 1 (A) . Concentration of IL-6 in collected medium was evaluated by standard ELISA method (B) ; Data are presented as mean ± standard deviation of duplicates from three independent assays and were analyzed with a Student's t -test (# P

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Inactive Gingipains from P. gingivalis Selectively Skews T Cells toward a Th17 Phenotype in an IL-6 Dependent Manner

    doi: 10.3389/fcimb.2017.00140

    Figure Lengend Snippet: The role of gingipains in the regulation of cell responses to LPS and fimbriae . Monocyte-derived dendritic cells (moDC) were untreated or exposed to P. gingivalis LPS (2.5 μg/ml), FimA (10 μg/ml) or mixed LPS with FimA. Additionally, cells were co-stimulated with HRgpA (2 nM) in the presence or absence of specific protease inhibitors (KYT-1 and KYT-36, each at a concentration of 1 μM). Four hours after stimulation, culture media were collected and cells were lysed with TRIzol. RNA was isolated and reverse transcriptase PCR was performed. Relative expression of the cytokine gene IL-6 to the reference house-keeping gene EF2 was measured by Real-Time PCR. Data show representative fold increase in expression compared to control levels, which were arbitrarily set at 1 (A) . Concentration of IL-6 in collected medium was evaluated by standard ELISA method (B) ; Data are presented as mean ± standard deviation of duplicates from three independent assays and were analyzed with a Student's t -test (# P

    Article Snippet: All characterized strains of P. gingivalis , as well as clinical isolates, produce gingipains (Ismail et al., ), which are major virulence factors capable of manipulation of a large number of defense and homeostatic mechanisms in infected tissue.

    Techniques: Derivative Assay, Concentration Assay, Isolation, Polymerase Chain Reaction, Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Standard Deviation

    Gingipain activity differentially determines the mRNA expression and final secretion of cytokines. (A) Monocyte-derived dendritic cells (moDC) were untreated or exposed to P. gingivalis W83 in the presence or absence of specific protease inhibitors (KYT-1 and KYT-36, each at a concentration of 1 μM) or the isogenic gingipain-null mutant ΔKΔRAB at MOI (multiplicity of infection) 1:50. (A) Concentrations of cytokines in collected medium were evaluated by ELISA (IL-6, IL-23, TNF, IFNγ) or by using BD CBA Human Inflammatory Kit (IL-1β, IL-12p70, IL-8, IL-10). A representative result from three separate experiments performed on moDCs derived from different donors is shown. Data are presented as mean ± standard deviation of duplicates from three independent assays and were analyzed with a Student's t -test (# P

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Inactive Gingipains from P. gingivalis Selectively Skews T Cells toward a Th17 Phenotype in an IL-6 Dependent Manner

    doi: 10.3389/fcimb.2017.00140

    Figure Lengend Snippet: Gingipain activity differentially determines the mRNA expression and final secretion of cytokines. (A) Monocyte-derived dendritic cells (moDC) were untreated or exposed to P. gingivalis W83 in the presence or absence of specific protease inhibitors (KYT-1 and KYT-36, each at a concentration of 1 μM) or the isogenic gingipain-null mutant ΔKΔRAB at MOI (multiplicity of infection) 1:50. (A) Concentrations of cytokines in collected medium were evaluated by ELISA (IL-6, IL-23, TNF, IFNγ) or by using BD CBA Human Inflammatory Kit (IL-1β, IL-12p70, IL-8, IL-10). A representative result from three separate experiments performed on moDCs derived from different donors is shown. Data are presented as mean ± standard deviation of duplicates from three independent assays and were analyzed with a Student's t -test (# P

    Article Snippet: All characterized strains of P. gingivalis , as well as clinical isolates, produce gingipains (Ismail et al., ), which are major virulence factors capable of manipulation of a large number of defense and homeostatic mechanisms in infected tissue.

    Techniques: Activity Assay, Expressing, Derivative Assay, Concentration Assay, Mutagenesis, Infection, Enzyme-linked Immunosorbent Assay, Crocin Bleaching Assay, Standard Deviation

    Th17 cells differentiation depends on gingipain activity . Monocyte-derived dendritic cells were untreated or exposed to P. gingivalis W83 in the presence/absence of specific protease inhibitors (KYT-1 and KYT-36, each at the concentration of 1 μM) for 6 h. Naive CD4 + cells were added and co-cultured for an additional 3 or 5 days. At the end of each period, T cells were stimulated with PMA and ionomycin for 4 h in the presence of GolgiStop. For evaluation of the Th17 population, cells were stain for intracellular IL-17A andRORγt, and were analyzed by flow cytometry. Data show a representative dot blot selected from three separate experiments presenting percent of RORγt + , IL-17+, RORγt + IL-17 + after 3 and 5 days of co-stimulation of naive CD4+ T cells with moDC pulsed with P. gingivalis .

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Inactive Gingipains from P. gingivalis Selectively Skews T Cells toward a Th17 Phenotype in an IL-6 Dependent Manner

    doi: 10.3389/fcimb.2017.00140

    Figure Lengend Snippet: Th17 cells differentiation depends on gingipain activity . Monocyte-derived dendritic cells were untreated or exposed to P. gingivalis W83 in the presence/absence of specific protease inhibitors (KYT-1 and KYT-36, each at the concentration of 1 μM) for 6 h. Naive CD4 + cells were added and co-cultured for an additional 3 or 5 days. At the end of each period, T cells were stimulated with PMA and ionomycin for 4 h in the presence of GolgiStop. For evaluation of the Th17 population, cells were stain for intracellular IL-17A andRORγt, and were analyzed by flow cytometry. Data show a representative dot blot selected from three separate experiments presenting percent of RORγt + , IL-17+, RORγt + IL-17 + after 3 and 5 days of co-stimulation of naive CD4+ T cells with moDC pulsed with P. gingivalis .

    Article Snippet: All characterized strains of P. gingivalis , as well as clinical isolates, produce gingipains (Ismail et al., ), which are major virulence factors capable of manipulation of a large number of defense and homeostatic mechanisms in infected tissue.

    Techniques: Activity Assay, Derivative Assay, Concentration Assay, Cell Culture, Staining, Flow Cytometry, Cytometry, Dot Blot

    Expression of interleukin 6 in human monocyte-derived dendritic cells in response to Porphyromonas gingivalis infection . Monocyte-derived dendritic cells (moDC) were untreated or exposed to P. gingivalis W83 in the presence or absence of specific protease inhibitors (KYT-1 and KYT-36, each at a concentration of 1 μM) or the isogenic gingipain-null mutant ΔKΔRAB for 2 h. After stimulation, cells were lysed with TRIzol, RNA was isolated and reverse transcriptase PCR was performed. Relative expression of IL-6 was measured by using the Real-Time PCR method. A representative qRT-PCR from three separate experiments performed on moDCs derived from different donors is shown. Data are presented as means ± standard deviations of triplicate assays, and were analyzed with the one-way ANOVA with the Bonferroni post-test correction (# P

    Journal: Frontiers in Cellular and Infection Microbiology

    Article Title: Inactive Gingipains from P. gingivalis Selectively Skews T Cells toward a Th17 Phenotype in an IL-6 Dependent Manner

    doi: 10.3389/fcimb.2017.00140

    Figure Lengend Snippet: Expression of interleukin 6 in human monocyte-derived dendritic cells in response to Porphyromonas gingivalis infection . Monocyte-derived dendritic cells (moDC) were untreated or exposed to P. gingivalis W83 in the presence or absence of specific protease inhibitors (KYT-1 and KYT-36, each at a concentration of 1 μM) or the isogenic gingipain-null mutant ΔKΔRAB for 2 h. After stimulation, cells were lysed with TRIzol, RNA was isolated and reverse transcriptase PCR was performed. Relative expression of IL-6 was measured by using the Real-Time PCR method. A representative qRT-PCR from three separate experiments performed on moDCs derived from different donors is shown. Data are presented as means ± standard deviations of triplicate assays, and were analyzed with the one-way ANOVA with the Bonferroni post-test correction (# P

    Article Snippet: All characterized strains of P. gingivalis , as well as clinical isolates, produce gingipains (Ismail et al., ), which are major virulence factors capable of manipulation of a large number of defense and homeostatic mechanisms in infected tissue.

    Techniques: Expressing, Derivative Assay, Infection, Concentration Assay, Mutagenesis, Isolation, Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

    IRAK1 expression was inhibited by miR-146a in B cells after challenged with P. gingivalis LPS miR-146a mimic could reduce the mRNA and protein levels of IRAK1 as detected by PCR (A) and ELISA (B). Also, addition of miR-146a inhibited TRAF6 expression in B cells at both mRNA (C) and protein level (D). Inhibition of miR-146a by its inhibitor significantly elevated both mRNA and protein levels of IRAK1 expression in B cells (E and F), as well as mRNA (G) and protein (H) levels of TRAF6 expression in B cells. The experiments were performed in triplicate. n=6–8. Data are presented as mean±SD. One-way ANOVA. * p

    Journal: Biochimica et biophysica acta

    Article Title: miR-146a regulates inflammatory cytokine production in Porphyromonas gingivalis lipopolysaccharide-stimulated B cells by targeting IRAK1 but not TRAF6

    doi: 10.1016/j.bbadis.2017.12.035

    Figure Lengend Snippet: IRAK1 expression was inhibited by miR-146a in B cells after challenged with P. gingivalis LPS miR-146a mimic could reduce the mRNA and protein levels of IRAK1 as detected by PCR (A) and ELISA (B). Also, addition of miR-146a inhibited TRAF6 expression in B cells at both mRNA (C) and protein level (D). Inhibition of miR-146a by its inhibitor significantly elevated both mRNA and protein levels of IRAK1 expression in B cells (E and F), as well as mRNA (G) and protein (H) levels of TRAF6 expression in B cells. The experiments were performed in triplicate. n=6–8. Data are presented as mean±SD. One-way ANOVA. * p

    Article Snippet: B cells were treated with miR-146a mimic, miR-146a inhibitor or scramble controls in the presence or absence of 1 μg/ml P. gingivalis LPS (Strain ATCC33277, InvivoGen, San Diego, CA, USA).

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

    The inflammatory responses in B cells were activated by P. gingivalis LPS After P. gingivalis LPS treatment, mRNA levels of inflammatory cytokines including IL-1β (A), IL-6 (B) and IL-10 (C) were significantly increased in B cells. IRAK1 mRNA level was also increased (D). However, TRAF6 mRNA level was not changed (E). The experiments were performed in triplicate. n=6–8. Data are presented as mean±SD. Student’s t-test. * p

    Journal: Biochimica et biophysica acta

    Article Title: miR-146a regulates inflammatory cytokine production in Porphyromonas gingivalis lipopolysaccharide-stimulated B cells by targeting IRAK1 but not TRAF6

    doi: 10.1016/j.bbadis.2017.12.035

    Figure Lengend Snippet: The inflammatory responses in B cells were activated by P. gingivalis LPS After P. gingivalis LPS treatment, mRNA levels of inflammatory cytokines including IL-1β (A), IL-6 (B) and IL-10 (C) were significantly increased in B cells. IRAK1 mRNA level was also increased (D). However, TRAF6 mRNA level was not changed (E). The experiments were performed in triplicate. n=6–8. Data are presented as mean±SD. Student’s t-test. * p

    Article Snippet: B cells were treated with miR-146a mimic, miR-146a inhibitor or scramble controls in the presence or absence of 1 μg/ml P. gingivalis LPS (Strain ATCC33277, InvivoGen, San Diego, CA, USA).

    Techniques:

    The expression of miR-146a in B cells was induced by P. gingivalis LPS The expression of miR-146a in B cells was detected by qRT-PCR after P. gingivalis LPS-stimulation. The miR-146a expression was increased at both 24 h and 48 h. The experiments were performed in triplicate. n=6–8. Data are presented as mean±SD. Student’s t-test. * p

    Journal: Biochimica et biophysica acta

    Article Title: miR-146a regulates inflammatory cytokine production in Porphyromonas gingivalis lipopolysaccharide-stimulated B cells by targeting IRAK1 but not TRAF6

    doi: 10.1016/j.bbadis.2017.12.035

    Figure Lengend Snippet: The expression of miR-146a in B cells was induced by P. gingivalis LPS The expression of miR-146a in B cells was detected by qRT-PCR after P. gingivalis LPS-stimulation. The miR-146a expression was increased at both 24 h and 48 h. The experiments were performed in triplicate. n=6–8. Data are presented as mean±SD. Student’s t-test. * p

    Article Snippet: B cells were treated with miR-146a mimic, miR-146a inhibitor or scramble controls in the presence or absence of 1 μg/ml P. gingivalis LPS (Strain ATCC33277, InvivoGen, San Diego, CA, USA).

    Techniques: Expressing, Quantitative RT-PCR

    The mRNA expressions of inflammatory cytokines were inhibited by miR-146a in B cells after challenged with P. gingivalis LPS After overexpression of miR-146a in B cells, mRNA levels of IL-1β (A), IL-6 (B) and IL-10 (C) were significantly decreased. However, after blockade of miR-146a by its inhibitor, the mRNA levels of IL-1β (D), IL-6 (E) and IL-10 (F) were significantly increased. The experiments were performed in triplicate. n=6–8. Data are presented as mean±SD. One-way ANOVA. * p

    Journal: Biochimica et biophysica acta

    Article Title: miR-146a regulates inflammatory cytokine production in Porphyromonas gingivalis lipopolysaccharide-stimulated B cells by targeting IRAK1 but not TRAF6

    doi: 10.1016/j.bbadis.2017.12.035

    Figure Lengend Snippet: The mRNA expressions of inflammatory cytokines were inhibited by miR-146a in B cells after challenged with P. gingivalis LPS After overexpression of miR-146a in B cells, mRNA levels of IL-1β (A), IL-6 (B) and IL-10 (C) were significantly decreased. However, after blockade of miR-146a by its inhibitor, the mRNA levels of IL-1β (D), IL-6 (E) and IL-10 (F) were significantly increased. The experiments were performed in triplicate. n=6–8. Data are presented as mean±SD. One-way ANOVA. * p

    Article Snippet: B cells were treated with miR-146a mimic, miR-146a inhibitor or scramble controls in the presence or absence of 1 μg/ml P. gingivalis LPS (Strain ATCC33277, InvivoGen, San Diego, CA, USA).

    Techniques: Over Expression

    miR-146a inhibited inflammatory cytokine secretion from in B cells after challenged with P. gingivalis LPS The secretions of IL-1β (A), IL-6 (B) and IL-10 (C) decreased in presence of miR-146a mimic. The productions of IL-1β (D), IL-6 (E) and IL-10 (F) were enhanced by miR-146a inhibitor. The experiments were performed in triplicate. n=6–8. Data are presented as mean±SD. One-way ANOVA. * p

    Journal: Biochimica et biophysica acta

    Article Title: miR-146a regulates inflammatory cytokine production in Porphyromonas gingivalis lipopolysaccharide-stimulated B cells by targeting IRAK1 but not TRAF6

    doi: 10.1016/j.bbadis.2017.12.035

    Figure Lengend Snippet: miR-146a inhibited inflammatory cytokine secretion from in B cells after challenged with P. gingivalis LPS The secretions of IL-1β (A), IL-6 (B) and IL-10 (C) decreased in presence of miR-146a mimic. The productions of IL-1β (D), IL-6 (E) and IL-10 (F) were enhanced by miR-146a inhibitor. The experiments were performed in triplicate. n=6–8. Data are presented as mean±SD. One-way ANOVA. * p

    Article Snippet: B cells were treated with miR-146a mimic, miR-146a inhibitor or scramble controls in the presence or absence of 1 μg/ml P. gingivalis LPS (Strain ATCC33277, InvivoGen, San Diego, CA, USA).

    Techniques: