p gingivalis  (ATCC)


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    Structured Review

    ATCC p gingivalis
    Attachment of T. forsythia 43037 (Tf) and BFM571 to KB cells in the absence and presence of P. <t>gingivalis</t> OMVs (Pgv). Attachment levels (means ± standard errors) were expressed as the percentage of attached bacteria relative to the total number
    P Gingivalis, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 37 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Porphyromonas gingivalis Vesicles Enhance Attachment, and the Leucine-Rich Repeat BspA Protein Is Required for Invasion of Epithelial Cells by "Tannerella forsythia" "

    Article Title: Porphyromonas gingivalis Vesicles Enhance Attachment, and the Leucine-Rich Repeat BspA Protein Is Required for Invasion of Epithelial Cells by "Tannerella forsythia"

    Journal:

    doi: 10.1128/IAI.00062-06

    Attachment of T. forsythia 43037 (Tf) and BFM571 to KB cells in the absence and presence of P. gingivalis OMVs (Pgv). Attachment levels (means ± standard errors) were expressed as the percentage of attached bacteria relative to the total number
    Figure Legend Snippet: Attachment of T. forsythia 43037 (Tf) and BFM571 to KB cells in the absence and presence of P. gingivalis OMVs (Pgv). Attachment levels (means ± standard errors) were expressed as the percentage of attached bacteria relative to the total number

    Techniques Used:

    Coaggregation of T. forsythia in the presence of P. gingivalis and outer membrane vesicles purified from P. gingivalis . Data are representative of three independent experiments. Data points represent means ± standard errors of triplicate readings.
    Figure Legend Snippet: Coaggregation of T. forsythia in the presence of P. gingivalis and outer membrane vesicles purified from P. gingivalis . Data are representative of three independent experiments. Data points represent means ± standard errors of triplicate readings.

    Techniques Used: Purification

    Invasion by T. forsythia (Tf) of KB cells and dependence of invasion on the presence of P. gingivalis vesicles (Pgv). The invasion level was expressed as the percentage of bacteria retrieved following antibiotic killing of external bacteria and KB cell
    Figure Legend Snippet: Invasion by T. forsythia (Tf) of KB cells and dependence of invasion on the presence of P. gingivalis vesicles (Pgv). The invasion level was expressed as the percentage of bacteria retrieved following antibiotic killing of external bacteria and KB cell

    Techniques Used:

    2) Product Images from "Effects of 1,25-dihydroxyvitamin D3 on experimental periodontitis and AhR/NF-κB/NLRP3 inflammasome pathway in a mouse model"

    Article Title: Effects of 1,25-dihydroxyvitamin D3 on experimental periodontitis and AhR/NF-κB/NLRP3 inflammasome pathway in a mouse model

    Journal: Journal of Applied Oral Science

    doi: 10.1590/1678-7757-2018-0713

    Gingival epithelial VDR expression in each group, and AhR, CYP1A1, p-p65, NLRP3, ASC, caspase-1, IL-1β and IL-6 expression in normal control group was shown in immunohistochemical images. Treatment with 1,25-dihydroxyvitamin D3 enhanced VDR expression, while the VDR expression exhibited no significant difference between normal control and untreated periodontitis mice. N, normal control; P, Porphyromonas gingivalis infection; V, Porphyromonas gingivalis infection with 1,25-dihydroxyvitamin D 3 treatment
    Figure Legend Snippet: Gingival epithelial VDR expression in each group, and AhR, CYP1A1, p-p65, NLRP3, ASC, caspase-1, IL-1β and IL-6 expression in normal control group was shown in immunohistochemical images. Treatment with 1,25-dihydroxyvitamin D3 enhanced VDR expression, while the VDR expression exhibited no significant difference between normal control and untreated periodontitis mice. N, normal control; P, Porphyromonas gingivalis infection; V, Porphyromonas gingivalis infection with 1,25-dihydroxyvitamin D 3 treatment

    Techniques Used: Expressing, Immunohistochemistry, Mouse Assay, Infection

    Protein expression of AhR, CYP1A1, p-p65, NLRP3, ASC, caspase-1, IL-1β and IL-6 in the mouse gingival epithelium was detected using immunohistochemical staining. After 1,25-dihydroxyvitamin D3 administration, the gingival epithelial expression of AhR and CYP1A1 was increased in mice with periodontitis, whereas the expression of p-p65, NLRP3, ASC, caspase-1, IL-1β and IL-6 was decreased. N, normal control; P, Porphyromonas gingivalis infection; V, Porphyromonas gingivalis infection with 1,25-dihydroxyvitamin D 3 treatment
    Figure Legend Snippet: Protein expression of AhR, CYP1A1, p-p65, NLRP3, ASC, caspase-1, IL-1β and IL-6 in the mouse gingival epithelium was detected using immunohistochemical staining. After 1,25-dihydroxyvitamin D3 administration, the gingival epithelial expression of AhR and CYP1A1 was increased in mice with periodontitis, whereas the expression of p-p65, NLRP3, ASC, caspase-1, IL-1β and IL-6 was decreased. N, normal control; P, Porphyromonas gingivalis infection; V, Porphyromonas gingivalis infection with 1,25-dihydroxyvitamin D 3 treatment

    Techniques Used: Expressing, Immunohistochemistry, Staining, Mouse Assay, Infection

    3) Product Images from "Interplay of Toll-Like Receptor 9, Myeloid Cells, and Deubiquitinase A20 in Periodontal Inflammation"

    Article Title: Interplay of Toll-Like Receptor 9, Myeloid Cells, and Deubiquitinase A20 in Periodontal Inflammation

    Journal: Infection and Immunity

    doi: 10.1128/IAI.00814-16

    Decreased inflammatory cell infiltrate in the peritoneal cavities of TLR9 −/− mice following Porphyromonas gingivalis challenge. WT and TLR9 −/− mice were intraperitoneally injected with 1 × 10 8 CFU of Porphyromonas
    Figure Legend Snippet: Decreased inflammatory cell infiltrate in the peritoneal cavities of TLR9 −/− mice following Porphyromonas gingivalis challenge. WT and TLR9 −/− mice were intraperitoneally injected with 1 × 10 8 CFU of Porphyromonas

    Techniques Used: Mouse Assay, Injection

    Loss of TLR9 in BMDM increased A20 (TNFAIP3) protein induction following P. gingivalis challenge. WT and TLR9 −/− BMDM were stimulated with heat-killed P. gingivalis (MOI of 1:100) for 2, 4, and 6 h. (A) mRNA was isolated, and expression
    Figure Legend Snippet: Loss of TLR9 in BMDM increased A20 (TNFAIP3) protein induction following P. gingivalis challenge. WT and TLR9 −/− BMDM were stimulated with heat-killed P. gingivalis (MOI of 1:100) for 2, 4, and 6 h. (A) mRNA was isolated, and expression

    Techniques Used: Isolation, Expressing

    4) Product Images from "Egg yolk immunoglobulin interactions with Porphyromonas gingivalis to impact periodontal inflammation and halitosis"

    Article Title: Egg yolk immunoglobulin interactions with Porphyromonas gingivalis to impact periodontal inflammation and halitosis

    Journal: AMB Express

    doi: 10.1186/s13568-018-0706-0

    Effect of IgY on rat gingivitis and bad breath induced by P. gingivalis . The scores for oral health index are shown as mean ± SD (n = 3). *Means different at p
    Figure Legend Snippet: Effect of IgY on rat gingivitis and bad breath induced by P. gingivalis . The scores for oral health index are shown as mean ± SD (n = 3). *Means different at p

    Techniques Used:

    The inhibitory effect of IgY on the growth of P. gingivalis . P. gingivalis was cultured in artificial saliva medium containing 0 (control), 0.0037 (low-dose), 7.4 (mid-dose) or 370 (high-dose) mmol/l of IgY. Data were obtained using three independent experiments with three measurements in each experiment, and are shown as mean ± SD (n = 3). *Means different at p
    Figure Legend Snippet: The inhibitory effect of IgY on the growth of P. gingivalis . P. gingivalis was cultured in artificial saliva medium containing 0 (control), 0.0037 (low-dose), 7.4 (mid-dose) or 370 (high-dose) mmol/l of IgY. Data were obtained using three independent experiments with three measurements in each experiment, and are shown as mean ± SD (n = 3). *Means different at p

    Techniques Used: Cell Culture

    5) Product Images from "Punica granatum L. (Pomegranate) Extract: In Vivo Study of Antimicrobial Activity against Porphyromonas gingivalis in Galleria mellonella Model"

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

    Journal: The Scientific World Journal

    doi: 10.1155/2016/8626987

    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.
    Figure Legend 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.

    Techniques Used: Infection

    6) Product Images from "In vitro Increased Respiratory Activity of Selected Oral Bacteria May Explain Competitive and Collaborative Interactions in the Oral Microbiome"

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

    Journal: Frontiers in Cellular and Infection Microbiology

    doi: 10.3389/fcimb.2017.00235

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

    Techniques Used: Activity Assay

    7) Product Images from "mTOR Inhibition Rejuvenates the Aging Gingival Fibroblasts through Alleviating Oxidative Stress"

    Article Title: mTOR Inhibition Rejuvenates the Aging Gingival Fibroblasts through Alleviating Oxidative Stress

    Journal: Oxidative Medicine and Cellular Longevity

    doi: 10.1155/2017/6292630

    Rapamycin enhances the anti-inflammatory ability of human gingival fibroblasts. (a) The results of real time-PCR show the mRNA expression of inflammatory cytokines IL-6 and IL-8. Both short- and long-term treatments of rapamycin can ease the inflammatory response caused by P. gingivalis infection. (b) FACS analysis shows the reactive oxygen species (ROS) levels in hGFs. Rapamycin treatment alleviates the intracellular ROS level in hGFs caused by P. gingivalis . (c) The results of real-time PCR show (right panel) the mRNA expression of antioxidant components Cat, Sod2, and Prdx3 (∗ means P
    Figure Legend Snippet: Rapamycin enhances the anti-inflammatory ability of human gingival fibroblasts. (a) The results of real time-PCR show the mRNA expression of inflammatory cytokines IL-6 and IL-8. Both short- and long-term treatments of rapamycin can ease the inflammatory response caused by P. gingivalis infection. (b) FACS analysis shows the reactive oxygen species (ROS) levels in hGFs. Rapamycin treatment alleviates the intracellular ROS level in hGFs caused by P. gingivalis . (c) The results of real-time PCR show (right panel) the mRNA expression of antioxidant components Cat, Sod2, and Prdx3 (∗ means P

    Techniques Used: Real-time Polymerase Chain Reaction, Expressing, Infection, FACS

    8) Product Images from "An early report: a modified porphyrin-linked metronidazole targeting intracellular Porphyromonas gingivalis in cultured oral epithelial cells"

    Article Title: An early report: a modified porphyrin-linked metronidazole targeting intracellular Porphyromonas gingivalis in cultured oral epithelial cells

    Journal: International Journal of Oral Science

    doi: 10.1038/ijos.2017.31

    Blood agar colonies (CFU of P. gingivalis ) that gave an indication of the efficacy of the porphyrin compound. ( a ) A negative control blood agar plate indicating no contamination. ( b ) Strong growth from the intracellular and adherent P. gingivalis culture for 1.5 h. ( c , d ) Invaded P. gingivalis treated with metronidazole (20 or 200 μg·mL −1 ) and gentamycin (30 or 300 μg·mL −1 ) for an additional 1 h, respectively, to compare whether there was any differences in killing adherent (extracellular) bacterial cells. Adherent/extracellular (surface-bound) bacteria were only eliminated with a combination of high levels of metronidazole (200 μg·mL −1 ) and gentamycin (300 μg·mL −1 ) ( d ). This represented intracellular infection of P. gingivalis. ( e , f ) Representative of cells treated with antibiotics and porphyrin compound or porphyrin compound only, for up to 4 and 6 h. All bacteria were eliminated. Panels ( g – i ) showed 6 h incubations with different treatments of P. gingivalis and antibiotics as controls. Bacteria were similar amounts with early time treatments ( b – d ), respectively. ( j ) The summary of the average counting numbers of intracellular P. gingivalis colonies (CFU) from blood agar cultures ( a – f ) at 4 days. CFU, colony forming units.
    Figure Legend Snippet: Blood agar colonies (CFU of P. gingivalis ) that gave an indication of the efficacy of the porphyrin compound. ( a ) A negative control blood agar plate indicating no contamination. ( b ) Strong growth from the intracellular and adherent P. gingivalis culture for 1.5 h. ( c , d ) Invaded P. gingivalis treated with metronidazole (20 or 200 μg·mL −1 ) and gentamycin (30 or 300 μg·mL −1 ) for an additional 1 h, respectively, to compare whether there was any differences in killing adherent (extracellular) bacterial cells. Adherent/extracellular (surface-bound) bacteria were only eliminated with a combination of high levels of metronidazole (200 μg·mL −1 ) and gentamycin (300 μg·mL −1 ) ( d ). This represented intracellular infection of P. gingivalis. ( e , f ) Representative of cells treated with antibiotics and porphyrin compound or porphyrin compound only, for up to 4 and 6 h. All bacteria were eliminated. Panels ( g – i ) showed 6 h incubations with different treatments of P. gingivalis and antibiotics as controls. Bacteria were similar amounts with early time treatments ( b – d ), respectively. ( j ) The summary of the average counting numbers of intracellular P. gingivalis colonies (CFU) from blood agar cultures ( a – f ) at 4 days. CFU, colony forming units.

    Techniques Used: Negative Control, Infection

    Live cell imaging of 3D reconstruction and colocalization of porphyrin adducts and pathogen. ( a ) Live cell imaging of z -stack projection at 3 h culture showed porphyrin-linked metronidazole adducts (in red, nuclei in blue) within the cytoplasm of epithelial H413-1 cells by confocal laser scanning microscopy. ( b ) 3D reconstruction with sliced images of one cell confirmed the compound (in red, nuclei in blue) localized in the cytoplasm. ( c ) Adherent and intracellular P. gingivalis shown at 1.5 h bacterial invasion to the cells (bacteria in green, nuclei in blue). ( d ) Intracellular P. gingivalis showing perinuclear spots in green (grey colour was a differential interference contrast (DIC)) after the killing of extracellular bacteria by combined antibiotics for additional 1 h. ( e ) Demonstration of the porphyrin adducts (in red) colocalized with P. gingivalis (in green, d ) within the cytoplasm around nuclei (perinuclear) of epithelial cells in yellow/orange colour ( f ) at additional 1.5 h culture of this compound.
    Figure Legend Snippet: Live cell imaging of 3D reconstruction and colocalization of porphyrin adducts and pathogen. ( a ) Live cell imaging of z -stack projection at 3 h culture showed porphyrin-linked metronidazole adducts (in red, nuclei in blue) within the cytoplasm of epithelial H413-1 cells by confocal laser scanning microscopy. ( b ) 3D reconstruction with sliced images of one cell confirmed the compound (in red, nuclei in blue) localized in the cytoplasm. ( c ) Adherent and intracellular P. gingivalis shown at 1.5 h bacterial invasion to the cells (bacteria in green, nuclei in blue). ( d ) Intracellular P. gingivalis showing perinuclear spots in green (grey colour was a differential interference contrast (DIC)) after the killing of extracellular bacteria by combined antibiotics for additional 1 h. ( e ) Demonstration of the porphyrin adducts (in red) colocalized with P. gingivalis (in green, d ) within the cytoplasm around nuclei (perinuclear) of epithelial cells in yellow/orange colour ( f ) at additional 1.5 h culture of this compound.

    Techniques Used: Live Cell Imaging, Confocal Laser Scanning Microscopy

    9) Product Images from "Interplay of Protease-Activated Receptors and NOD Pattern Recognition Receptors in Epithelial Innate Immune Responses to Bacteria"

    Article Title: Interplay of Protease-Activated Receptors and NOD Pattern Recognition Receptors in Epithelial Innate Immune Responses to Bacteria

    Journal: Immunology letters

    doi: 10.1016/j.imlet.2010.02.006

    The effect of silencing PAR1 (P1) or PAR2 (P2) on IL-8 induction in response to various oral bacteria is evaluated by ELISA. Cells were transfected with a specific siRNA for 48 h, then subsequently stimulated with bacteria for 16 h. Cell-free supernatant was collected and the amount of IL-8 secreted is shown in pg/ml. Unstimulated cells (UN) are used as controls in each experiment. Data from duplicates with cells from three different donors are shown. PG: P. gingivalis ; FN: F. nucleatum ; SG: S. gordonii .
    Figure Legend Snippet: The effect of silencing PAR1 (P1) or PAR2 (P2) on IL-8 induction in response to various oral bacteria is evaluated by ELISA. Cells were transfected with a specific siRNA for 48 h, then subsequently stimulated with bacteria for 16 h. Cell-free supernatant was collected and the amount of IL-8 secreted is shown in pg/ml. Unstimulated cells (UN) are used as controls in each experiment. Data from duplicates with cells from three different donors are shown. PG: P. gingivalis ; FN: F. nucleatum ; SG: S. gordonii .

    Techniques Used: Enzyme-linked Immunosorbent Assay, Transfection

    QRT-PCR analyses of the effect of silencing PAR1 or PAR2 on NOD1 (A) and NOD2 (B) mRNA induction by various oral bacteria. Cells were transfected with a specific siRNA for 48 h, then subsequently stimulated with bacteria for 16 h. GECs transfected with non-silencing (NS) siRNA and unstimulated cells (UN) are used as controls in each experiment. Data from duplicates with cells from three different donors were normalized to the housekeeping gene control ribosomal phosphoprotein, and the average with standard deviation is shown. PG: P. gingivalis ; FN: F. nucleatum ; SG: S. gordonii .
    Figure Legend Snippet: QRT-PCR analyses of the effect of silencing PAR1 or PAR2 on NOD1 (A) and NOD2 (B) mRNA induction by various oral bacteria. Cells were transfected with a specific siRNA for 48 h, then subsequently stimulated with bacteria for 16 h. GECs transfected with non-silencing (NS) siRNA and unstimulated cells (UN) are used as controls in each experiment. Data from duplicates with cells from three different donors were normalized to the housekeeping gene control ribosomal phosphoprotein, and the average with standard deviation is shown. PG: P. gingivalis ; FN: F. nucleatum ; SG: S. gordonii .

    Techniques Used: Quantitative RT-PCR, Transfection, Standard Deviation

    QRT-PCR analyses of the effect of silencing PAR1 or PAR2 on TLR2 (A) and TLR4 (B) mRNA induction by various oral bacteria. Cells were transfected with a specific siRNA for 48 h, then subsequently stimulated with bacteria for 16 h. GECs transfected with non-silencing (NS) siRNA and unstimulated cells (UN) are used as controls in each experiment. Data from duplicates with cells from three different donors were normalized to the housekeeping gene control ribosomal phosphoprotein, and the average with standard deviation is shown. PG: P. gingivalis ; FN: F. nucleatum ; SG: S. gordonii .
    Figure Legend Snippet: QRT-PCR analyses of the effect of silencing PAR1 or PAR2 on TLR2 (A) and TLR4 (B) mRNA induction by various oral bacteria. Cells were transfected with a specific siRNA for 48 h, then subsequently stimulated with bacteria for 16 h. GECs transfected with non-silencing (NS) siRNA and unstimulated cells (UN) are used as controls in each experiment. Data from duplicates with cells from three different donors were normalized to the housekeeping gene control ribosomal phosphoprotein, and the average with standard deviation is shown. PG: P. gingivalis ; FN: F. nucleatum ; SG: S. gordonii .

    Techniques Used: Quantitative RT-PCR, Transfection, Standard Deviation

    QRT-PCR analyses of the effect of silencing PAR1 or PAR2 on CCL20 (A) and hBD-2 (B) mRNA induction by various oral bacteria. Cells were transfected with a specific siRNA for 48 h, then subsequently stimulated with bacteria for 16 h. GECs transfected with non-silencing (NS) siRNA and unstimulated cells (UN) are used as controls in each experiment. Data from duplicates with cells from three different donors were normalized to the housekeeping gene control ribosomal phosphoprotein, and the average with standard deviation is shown. PG: P. gingivalis ; FN: F. nucleatum ; SG: S. gordonii .
    Figure Legend Snippet: QRT-PCR analyses of the effect of silencing PAR1 or PAR2 on CCL20 (A) and hBD-2 (B) mRNA induction by various oral bacteria. Cells were transfected with a specific siRNA for 48 h, then subsequently stimulated with bacteria for 16 h. GECs transfected with non-silencing (NS) siRNA and unstimulated cells (UN) are used as controls in each experiment. Data from duplicates with cells from three different donors were normalized to the housekeeping gene control ribosomal phosphoprotein, and the average with standard deviation is shown. PG: P. gingivalis ; FN: F. nucleatum ; SG: S. gordonii .

    Techniques Used: Quantitative RT-PCR, Transfection, Standard Deviation

    The effect of silencing PAR1 (P1) or PAR2 (P2) on the induction of multiple cytokines in response to various oral bacteria is evaluated by ELISA. Cells were transfected with a specific siRNA for 48 h, then subsequently stimulated with bacteria for 16 h. Cell-free supernatant was collected and the amount of cytokines secreted is compared in fold changes relative to bacteria only controls. Data from two different donor cells are shown. PG: P. gingivalis ; FN: F. nucleatum ; SG: S. gordonii .
    Figure Legend Snippet: The effect of silencing PAR1 (P1) or PAR2 (P2) on the induction of multiple cytokines in response to various oral bacteria is evaluated by ELISA. Cells were transfected with a specific siRNA for 48 h, then subsequently stimulated with bacteria for 16 h. Cell-free supernatant was collected and the amount of cytokines secreted is compared in fold changes relative to bacteria only controls. Data from two different donor cells are shown. PG: P. gingivalis ; FN: F. nucleatum ; SG: S. gordonii .

    Techniques Used: Enzyme-linked Immunosorbent Assay, Transfection

    10) Product Images from "Serine Lipids of Porphyromonas gingivalis Are Human and Mouse Toll-Like Receptor 2 Ligands"

    Article Title: Serine Lipids of Porphyromonas gingivalis Are Human and Mouse Toll-Like Receptor 2 Ligands

    Journal: Infection and Immunity

    doi: 10.1128/IAI.00803-13

    HEK293 activation by lipids recovered in HPLC fractions of P. gingivalis total lipids. HEK293 cells, transfected with the human TLR2, MD-2, CD14, and SEAP genes, were used to assay the functions of P. gingivalis lipid fractions in vitro . A defined volume of each HPLC fraction was dried and reconstituted in 50% DMSO in water. The final concentration of DMSO achieved in culture medium was 1.11%. HEK293 cells were stimulated for 24 h with a defined amount of each lipid fraction. Results are expressed as the stimulated/nonstimulated (DMSO control) response ratio of HEK293 cells (graph A). Graphs B and C show the ion abundances within each HPLC fraction of lipids that produced negative ions of m/z 654 and 430, respectively.
    Figure Legend Snippet: HEK293 activation by lipids recovered in HPLC fractions of P. gingivalis total lipids. HEK293 cells, transfected with the human TLR2, MD-2, CD14, and SEAP genes, were used to assay the functions of P. gingivalis lipid fractions in vitro . A defined volume of each HPLC fraction was dried and reconstituted in 50% DMSO in water. The final concentration of DMSO achieved in culture medium was 1.11%. HEK293 cells were stimulated for 24 h with a defined amount of each lipid fraction. Results are expressed as the stimulated/nonstimulated (DMSO control) response ratio of HEK293 cells (graph A). Graphs B and C show the ion abundances within each HPLC fraction of lipids that produced negative ions of m/z 654 and 430, respectively.

    Techniques Used: Activation Assay, High Performance Liquid Chromatography, Transfection, In Vitro, Concentration Assay, Produced

    HEK cell activation by lipid classes derived from P. gingivalis . HEK293 cells, transfected with the human TLR2, MD-2, CD14, and SEAP genes, were used to assay the function of P. gingivalis lipid classes in vitro . HEK293 cells were stimulated for 24 h with the following treatments: DMSO (vehicle; 50% mixture of DMSO-water, approximately 1.11% DMSO in the final culture medium; n = 23), the known TLR2 ligand MMP (0.2 μg/ml; n = 34), LTA (2 μg/ml; n = 17), lipid 654 (0.17 μg/ml [ n = 5]; 0.34 μg/ml [ n = 2]; 0.69 μg/ml [ n = 22]), lipid 430 (0.17 μg/ml [ n = 2]; 0.34 μg/ml [ n = 3]; 0.69 μg/ml [ n = 18]), subPG-DHC (0.69 μg/ml; n = 4), unPG-DHC (0.69 μg/ml; n = 4), PE-DHC (0.69 μg/ml; n = 4), and PEA (0.69 μg/ml; n = 9). The phospholipid preparations were prepared from P. gingivalis ). Responses were assessed after 24 h, and results are expressed as the ratio of stimulated versus nonstimulated (DMSO) responses. By one-way ANOVA and Fisher LSD pairwise comparisons, HEK cell activation levels by MMP, LTA, lipid 654, and lipid 430 (both at 0.69 μg/ml) were significantly elevated over the DMSO vehicle ( P
    Figure Legend Snippet: HEK cell activation by lipid classes derived from P. gingivalis . HEK293 cells, transfected with the human TLR2, MD-2, CD14, and SEAP genes, were used to assay the function of P. gingivalis lipid classes in vitro . HEK293 cells were stimulated for 24 h with the following treatments: DMSO (vehicle; 50% mixture of DMSO-water, approximately 1.11% DMSO in the final culture medium; n = 23), the known TLR2 ligand MMP (0.2 μg/ml; n = 34), LTA (2 μg/ml; n = 17), lipid 654 (0.17 μg/ml [ n = 5]; 0.34 μg/ml [ n = 2]; 0.69 μg/ml [ n = 22]), lipid 430 (0.17 μg/ml [ n = 2]; 0.34 μg/ml [ n = 3]; 0.69 μg/ml [ n = 18]), subPG-DHC (0.69 μg/ml; n = 4), unPG-DHC (0.69 μg/ml; n = 4), PE-DHC (0.69 μg/ml; n = 4), and PEA (0.69 μg/ml; n = 9). The phospholipid preparations were prepared from P. gingivalis ). Responses were assessed after 24 h, and results are expressed as the ratio of stimulated versus nonstimulated (DMSO) responses. By one-way ANOVA and Fisher LSD pairwise comparisons, HEK cell activation levels by MMP, LTA, lipid 654, and lipid 430 (both at 0.69 μg/ml) were significantly elevated over the DMSO vehicle ( P

    Techniques Used: Activation Assay, Derivative Assay, Transfection, In Vitro

    Lipid 654 and lipid 430 are not agonists for TLR4. HEK293 cells, transfected with the human TLR4, CD14, MD-2, and SEAP genes, were used to assay the function of lipid 654 and lipid 430 in vitro . HEK293 cells were stimulated for 24 h with DMSO (vehicle; 50% DMSO in water), the known TLR2 ligands MMP and LTA, lipid 654 (0.69 μg/ml), lipid 430 (0.69 μg/ml), and two different preparations of known TLR4 agonists, LPS derived from Salmonella enterica or P. gingivalis (1 μg/ml). Responses were assessed at 24 h, and results are expressed as the ratio of stimulated to nonstimulated (DMSO) responses ( n = 2).
    Figure Legend Snippet: Lipid 654 and lipid 430 are not agonists for TLR4. HEK293 cells, transfected with the human TLR4, CD14, MD-2, and SEAP genes, were used to assay the function of lipid 654 and lipid 430 in vitro . HEK293 cells were stimulated for 24 h with DMSO (vehicle; 50% DMSO in water), the known TLR2 ligands MMP and LTA, lipid 654 (0.69 μg/ml), lipid 430 (0.69 μg/ml), and two different preparations of known TLR4 agonists, LPS derived from Salmonella enterica or P. gingivalis (1 μg/ml). Responses were assessed at 24 h, and results are expressed as the ratio of stimulated to nonstimulated (DMSO) responses ( n = 2).

    Techniques Used: Transfection, In Vitro, Derivative Assay

    11) Product Images from "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"

    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

    Journal: Infection and Immunity

    doi: 10.1128/IAI.00498-15

    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.
    Figure Legend 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.

    Techniques Used: 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
    Figure Legend 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

    Techniques Used: Real-time Polymerase Chain Reaction

    P. gingivalis regulates Angpt2 through ETS1 in AoSMCs. AoSMCs were treated with (KD) or without (Control) ETS1 siRNA or nontargeting siRNA (NT). The cells were then infected or not infected with P. gingivalis W50 for 24 h. (A and B) Quantitative real-time
    Figure Legend Snippet: P. gingivalis regulates Angpt2 through ETS1 in AoSMCs. AoSMCs were treated with (KD) or without (Control) ETS1 siRNA or nontargeting siRNA (NT). The cells were then infected or not infected with P. gingivalis W50 for 24 h. (A and B) Quantitative real-time

    Techniques Used: Infection

    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
    Figure Legend 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

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction

    12) Product Images from "Looking in the Porphyromonas gingivalis’ Cabinet of Curiosities: The Microbium, the Host and Cancer Association"

    Article Title: Looking in the Porphyromonas gingivalis’ Cabinet of Curiosities: The Microbium, the Host and Cancer Association

    Journal: Molecular oral microbiology

    doi: 10.1111/omi.12047

    Postulated pro-cancer molecular circuitry of P. gingivalis and OECs interface that may affect the increased risk of orodigestive cancers and their poor prognosis. Over the period of its co-evolution with human oral mucosa, P. gingivalis has established
    Figure Legend Snippet: Postulated pro-cancer molecular circuitry of P. gingivalis and OECs interface that may affect the increased risk of orodigestive cancers and their poor prognosis. Over the period of its co-evolution with human oral mucosa, P. gingivalis has established

    Techniques Used:

    13) Product Images from "Porphyromonas gingivalis in Alzheimer’s disease brains: Evidence for disease causation and treatment with small-molecule inhibitors"

    Article Title: Porphyromonas gingivalis in Alzheimer’s disease brains: Evidence for disease causation and treatment with small-molecule inhibitors

    Journal: Science Advances

    doi: 10.1126/sciadv.aau3333

    COR388 target engagement and dose-dependent effects on brain P. gingivalis , Aβ 1–42 , and TNFα in mice. ( A ) COR553 fluorescent activity probe for Kgp. ( B ) COR553 labeling of Kgp in P. gingivalis W83 strain and no labeling in mutant deficient in Kgp (ΔKgp). ( C ) W83 lysates labeled with COR553. Left lane, before immunodepletion; middle lane, after immunodepletion with anti-Kgp–conjugated beads; right lane, after elution from anti-Kgp–conjugated beads. ( D ) W83 strain titrated and labeled with COR553 to determine the limit of bacterial detection. See Results for details. ( E ) Oral plaque samples from human subjects (CB1-5) with periodontal disease were incubated ex vivo with COR553 probe with or without preincubation with COR388. COR553 probe and CAB102 detected Kgp strongly in three subjects (CB1, CB4, and CB5) and weakly in one subject (CB3). COR388 preincubation blocked COR553 probe binding to Kgp. ( F ) qPCR analysis of plaque samples using hmuY gene–specific primers identified P. gingivalis DNA in samples. ( G ) qPCR analysis of saliva samples. The bar graphs in (F) and (G) show the means and SEMs of three replicates. ( H ) COR388 treatment of W83 culture in defined growth medium reduced growth similarly to a Kgp-deficient strain (ΔKgp) over 43 hours. ( I ) Resistance developed rapidly to moxifloxacin but not COR388 with repeat passaging of bacterial culture. ( J to L ) Efficacy of COR388 at three oral doses of 3, 10, and 30 mg/kg twice daily in treating an established P. gingivalis brain infection in mice. Reduction of brain tissue levels of P. gingivalis (J), Aβ 1–42 (K), and TNFα (L). The bar graphs show the means with SEM error bars. *** P
    Figure Legend Snippet: COR388 target engagement and dose-dependent effects on brain P. gingivalis , Aβ 1–42 , and TNFα in mice. ( A ) COR553 fluorescent activity probe for Kgp. ( B ) COR553 labeling of Kgp in P. gingivalis W83 strain and no labeling in mutant deficient in Kgp (ΔKgp). ( C ) W83 lysates labeled with COR553. Left lane, before immunodepletion; middle lane, after immunodepletion with anti-Kgp–conjugated beads; right lane, after elution from anti-Kgp–conjugated beads. ( D ) W83 strain titrated and labeled with COR553 to determine the limit of bacterial detection. See Results for details. ( E ) Oral plaque samples from human subjects (CB1-5) with periodontal disease were incubated ex vivo with COR553 probe with or without preincubation with COR388. COR553 probe and CAB102 detected Kgp strongly in three subjects (CB1, CB4, and CB5) and weakly in one subject (CB3). COR388 preincubation blocked COR553 probe binding to Kgp. ( F ) qPCR analysis of plaque samples using hmuY gene–specific primers identified P. gingivalis DNA in samples. ( G ) qPCR analysis of saliva samples. The bar graphs in (F) and (G) show the means and SEMs of three replicates. ( H ) COR388 treatment of W83 culture in defined growth medium reduced growth similarly to a Kgp-deficient strain (ΔKgp) over 43 hours. ( I ) Resistance developed rapidly to moxifloxacin but not COR388 with repeat passaging of bacterial culture. ( J to L ) Efficacy of COR388 at three oral doses of 3, 10, and 30 mg/kg twice daily in treating an established P. gingivalis brain infection in mice. Reduction of brain tissue levels of P. gingivalis (J), Aβ 1–42 (K), and TNFα (L). The bar graphs show the means with SEM error bars. *** P

    Techniques Used: Mouse Assay, Activity Assay, Labeling, Mutagenesis, Incubation, Ex Vivo, Binding Assay, Real-time Polymerase Chain Reaction, Passaging, Infection

    Small-molecule gingipain inhibitors protect neuronal cells against P. gingivalis – and gingipain-induced toxicity in vitro and in vivo. ( A ) Differentiated SH-SY5Y neuroblastoma cells demonstrate cell aggregation after exposure to RgpB (10 μg/ml), Kgp (10 μg/ml), or both for 24 hours. The nonselective cysteine protease inhibitor iodoacetamide (IAM) blocks the gingipain-induced cell aggregation. ( B ) AlamarBlue viability assay shows that P. gingivalis ( P.g. ) is toxic to SH-SY5Y cells (MOI of 400) and that the small-molecule Kgp inhibitor COR271 and the RgpB inhibitor COR286 provide dose-dependent protection. The broad-spectrum antibiotics moxifloxacin and doxycycline and the γ-secretase inhibitor semagacestat did not inhibit the cytotoxic effect of P. gingivalis . ( C ) Fluoro-Jade C (FJC) staining (green) in pyramidal neurons of the CA1 region of the mouse hippocampus indicates neurodegeneration after stereotactic injection of gingipains. Counterstain with 4′,6-diamidino-2-phenylindole (DAPI) (blue). Scale bars, 50 μm. ( D ) The total number of FJC-positive cells was determined from serial section through the entire hippocampus. Results demonstrate a significant neuroprotective effect of gingipain inhibitors COR271 + COR286 after acute gingipain exposure in the hippocampus (* P
    Figure Legend Snippet: Small-molecule gingipain inhibitors protect neuronal cells against P. gingivalis – and gingipain-induced toxicity in vitro and in vivo. ( A ) Differentiated SH-SY5Y neuroblastoma cells demonstrate cell aggregation after exposure to RgpB (10 μg/ml), Kgp (10 μg/ml), or both for 24 hours. The nonselective cysteine protease inhibitor iodoacetamide (IAM) blocks the gingipain-induced cell aggregation. ( B ) AlamarBlue viability assay shows that P. gingivalis ( P.g. ) is toxic to SH-SY5Y cells (MOI of 400) and that the small-molecule Kgp inhibitor COR271 and the RgpB inhibitor COR286 provide dose-dependent protection. The broad-spectrum antibiotics moxifloxacin and doxycycline and the γ-secretase inhibitor semagacestat did not inhibit the cytotoxic effect of P. gingivalis . ( C ) Fluoro-Jade C (FJC) staining (green) in pyramidal neurons of the CA1 region of the mouse hippocampus indicates neurodegeneration after stereotactic injection of gingipains. Counterstain with 4′,6-diamidino-2-phenylindole (DAPI) (blue). Scale bars, 50 μm. ( D ) The total number of FJC-positive cells was determined from serial section through the entire hippocampus. Results demonstrate a significant neuroprotective effect of gingipain inhibitors COR271 + COR286 after acute gingipain exposure in the hippocampus (* P

    Techniques Used: In Vitro, In Vivo, Protease Inhibitor, Viability Assay, Staining, Injection

    RgpB colocalizes with neurons and pathology in AD hippocampus. ( A ) IHC using RgpB-specific monoclonal antibody 18E6 (representative images from a 63-year-old AD patient). The hippocampus shows abundant intracellular RgpB in the hilus (1), CA3 pyramidal layer (2), granular cell layer (3), and molecular layer (4). High-magnification images from the indicated areas (1 to 4) exhibit a granular staining pattern consistent with P. gingivalis intracellular infection. Scale bars, 200 μm (overview), 50 μm (1), and 10 μm (2 to 4). ( B ) AD hippocampus stained with 18E6 (AD) compared to gingival tissue (gingiva) from a patient with periodontal disease as well as a non-AD control and mouse IgG1 control (IgG1) in an adjacent hippocampal section. Scale bars, 50 μm. ( C ) Immunofluorescent colabeling with CAB101 reveals granular intraneuronal staining for RgpB (arrows) in MAP2-positive neurons in both the granular cell layer (GCL) and the pyramidal cell layer (CA1). Scale bars, 10 μm. ( D ) Dense extracellular RgpB-positive aggregates (arrowheads) were closely associated with astrocytes [glial fibrillary acidic protein (GFAP)]. There was no observed association of RgpB with microglia (IBA1). Scale bars, 10 μm. ( E ) RgpB was associated with paired helical filament Tau (PHF-Tau; arrows). RgpB-positive neurons negative for PHF-Tau (arrowheads) were also seen. Intracellular Aβ was often colocalized with RgpB (arrows). In some Aβ-positive cells, RgpB could not be detected (arrowheads). Scale bars, 10 μm.
    Figure Legend Snippet: RgpB colocalizes with neurons and pathology in AD hippocampus. ( A ) IHC using RgpB-specific monoclonal antibody 18E6 (representative images from a 63-year-old AD patient). The hippocampus shows abundant intracellular RgpB in the hilus (1), CA3 pyramidal layer (2), granular cell layer (3), and molecular layer (4). High-magnification images from the indicated areas (1 to 4) exhibit a granular staining pattern consistent with P. gingivalis intracellular infection. Scale bars, 200 μm (overview), 50 μm (1), and 10 μm (2 to 4). ( B ) AD hippocampus stained with 18E6 (AD) compared to gingival tissue (gingiva) from a patient with periodontal disease as well as a non-AD control and mouse IgG1 control (IgG1) in an adjacent hippocampal section. Scale bars, 50 μm. ( C ) Immunofluorescent colabeling with CAB101 reveals granular intraneuronal staining for RgpB (arrows) in MAP2-positive neurons in both the granular cell layer (GCL) and the pyramidal cell layer (CA1). Scale bars, 10 μm. ( D ) Dense extracellular RgpB-positive aggregates (arrowheads) were closely associated with astrocytes [glial fibrillary acidic protein (GFAP)]. There was no observed association of RgpB with microglia (IBA1). Scale bars, 10 μm. ( E ) RgpB was associated with paired helical filament Tau (PHF-Tau; arrows). RgpB-positive neurons negative for PHF-Tau (arrowheads) were also seen. Intracellular Aβ was often colocalized with RgpB (arrows). In some Aβ-positive cells, RgpB could not be detected (arrowheads). Scale bars, 10 μm.

    Techniques Used: Immunohistochemistry, Staining, Infection

    Identification of P. gingivalis –specific protein and DNA in cortex from control and AD patients. ( A ) WB with four different strains of P. gingivalis and CAB102 detection of typical molecular weight bands for Kgp in bacterial lysates. ( B ) IP using brain lysates from nondemented controls (C1 to C6; ages 75, 54, 63, 45, 37, and 102 years, respectively) and AD patients (AD1 to AD3; ages 83, 90, and 80 years, respectively) using CAB102 with subsequent WB reveals the ~50-kDa Kgp catalytic subunit (Kgp cat ), along with higher– and lower–molecular weight Kgp species seen in (A). ( C ) qPCR from DNA isolated from the same brain lysates as the protein samples analyzed in (B) shows a positive signal in nondemented control (C1 to C5) and AD (AD1 to AD3) samples. Sample C6 from the 102-year-old nondemented control patient had no detectable qPCR signal in (C) and very faint bands indicating near absence of Kgp (B) (mean with SEM error bars of repeat qPCR runs).
    Figure Legend Snippet: Identification of P. gingivalis –specific protein and DNA in cortex from control and AD patients. ( A ) WB with four different strains of P. gingivalis and CAB102 detection of typical molecular weight bands for Kgp in bacterial lysates. ( B ) IP using brain lysates from nondemented controls (C1 to C6; ages 75, 54, 63, 45, 37, and 102 years, respectively) and AD patients (AD1 to AD3; ages 83, 90, and 80 years, respectively) using CAB102 with subsequent WB reveals the ~50-kDa Kgp catalytic subunit (Kgp cat ), along with higher– and lower–molecular weight Kgp species seen in (A). ( C ) qPCR from DNA isolated from the same brain lysates as the protein samples analyzed in (B) shows a positive signal in nondemented control (C1 to C5) and AD (AD1 to AD3) samples. Sample C6 from the 102-year-old nondemented control patient had no detectable qPCR signal in (C) and very faint bands indicating near absence of Kgp (B) (mean with SEM error bars of repeat qPCR runs).

    Techniques Used: Western Blot, Molecular Weight, Real-time Polymerase Chain Reaction, Isolation

    Detection of P. gingivalis in CSF and oral biofluids from clinical AD subjects. ( A ) Detection and quantitation of P. gingivalis DNA by qPCR in CSF from subjects with probable AD. ( B ) Detection and quantitation of P. gingivalis DNA by qPCR from matching saliva samples. ( C ) Top: PCR products detecting P. gingivalis from CSF in (A) from all subjects run on agarose gel including negative and positive controls containing a synthetic DNA template. Faint or undetectable PCR products from subjects AD1, AD3, and AD5 were below the limit of quantitation for copy number and not of sufficient quantity for sequence analysis. Bottom: qPCR products from CSF from the same subjects for H. pylori. ( D ) Data table includes age and Mini Mental Status Exam (MMSE) score on subjects and sequence identity of PCR products to P. gingivalis hmuY DNA sequence. Sequence data are included in fig. S4. NS, not sequenced.
    Figure Legend Snippet: Detection of P. gingivalis in CSF and oral biofluids from clinical AD subjects. ( A ) Detection and quantitation of P. gingivalis DNA by qPCR in CSF from subjects with probable AD. ( B ) Detection and quantitation of P. gingivalis DNA by qPCR from matching saliva samples. ( C ) Top: PCR products detecting P. gingivalis from CSF in (A) from all subjects run on agarose gel including negative and positive controls containing a synthetic DNA template. Faint or undetectable PCR products from subjects AD1, AD3, and AD5 were below the limit of quantitation for copy number and not of sufficient quantity for sequence analysis. Bottom: qPCR products from CSF from the same subjects for H. pylori. ( D ) Data table includes age and Mini Mental Status Exam (MMSE) score on subjects and sequence identity of PCR products to P. gingivalis hmuY DNA sequence. Sequence data are included in fig. S4. NS, not sequenced.

    Techniques Used: Quantitation Assay, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Sequencing

    P. gingivalis invasion of the brain induces an Aβ 1–42 response that is blocked by gingipain inhibition in mice. ( A ) P. gingivalis PCR product in mouse brains after oral infection with P. gingivalis W83, with or without treatment with the Kgp inhibitor COR119, or infection with gingipain knockout strain ΔRgpB or ΔKgp. Lanes 1 to 8 represent individual experimental animals. In the first lane ( P.g. ), P. gingivalis W83 was used as a positive control. ( B ) P. gingivalis W83–infected mice, but not COR119-treated mice or mice infected with gingipain knockouts, had significantly higher Aβ 1–42 levels compared to mock-infected mice (*** P
    Figure Legend Snippet: P. gingivalis invasion of the brain induces an Aβ 1–42 response that is blocked by gingipain inhibition in mice. ( A ) P. gingivalis PCR product in mouse brains after oral infection with P. gingivalis W83, with or without treatment with the Kgp inhibitor COR119, or infection with gingipain knockout strain ΔRgpB or ΔKgp. Lanes 1 to 8 represent individual experimental animals. In the first lane ( P.g. ), P. gingivalis W83 was used as a positive control. ( B ) P. gingivalis W83–infected mice, but not COR119-treated mice or mice infected with gingipain knockouts, had significantly higher Aβ 1–42 levels compared to mock-infected mice (*** P

    Techniques Used: Inhibition, Mouse Assay, Polymerase Chain Reaction, Infection, Knock-Out, Positive Control

    P. gingivalis and gingipains fragment tau. ( A ) WB analysis of total soluble tau in SH-SY5Y cells infected with increasing concentrations of wild-type (WT) P. gingivalis strain W83 ( P.g. ) and P. gingivalis gingipain-deficient mutants either lacking Kgp activity (KgpΔIg-B) or lacking both Kgp and Rgp activity (ΔK/ΔRAB-A) . Uninfected SH-SY5Y cells (No P.g. ) were used as a negative control. Glyceraldehyde-phosphate dehydrogenase (GAPDH) was used as a loading control. Total tau was monitored with the monoclonal antibody Tau-5 at 1, 4, and 8 hours after infection. ( B ) Densitometry analysis of the total tau WB images. ( C ) WB analysis of rtau-441 incubated with purified Kgp and RgpB catalytic domains combined (Gp) at various concentrations for 1 hour at 37°C. The blot was probed with tau monoclonal antibody T46. ( D ) Gingipain cleavage sites in rtau-441 deduced from peptide fragments identified by MS for rtau-441 incubated with 1 or 10 nM gingipains. (a) T46 antibody epitope (red). (b) Tau-5 antibody epitope (red). (c) N-terminal tau fragment. (d) C-terminal tau fragment. (e) Kgp-generated tau fragments containing the VQIVYK sequence. (f) Kgp-generated fragments containing the VQIINK sequence. (g) An RgpB-generated tau fragment. *Cleavage sites identified at 1 nM gingipains.
    Figure Legend Snippet: P. gingivalis and gingipains fragment tau. ( A ) WB analysis of total soluble tau in SH-SY5Y cells infected with increasing concentrations of wild-type (WT) P. gingivalis strain W83 ( P.g. ) and P. gingivalis gingipain-deficient mutants either lacking Kgp activity (KgpΔIg-B) or lacking both Kgp and Rgp activity (ΔK/ΔRAB-A) . Uninfected SH-SY5Y cells (No P.g. ) were used as a negative control. Glyceraldehyde-phosphate dehydrogenase (GAPDH) was used as a loading control. Total tau was monitored with the monoclonal antibody Tau-5 at 1, 4, and 8 hours after infection. ( B ) Densitometry analysis of the total tau WB images. ( C ) WB analysis of rtau-441 incubated with purified Kgp and RgpB catalytic domains combined (Gp) at various concentrations for 1 hour at 37°C. The blot was probed with tau monoclonal antibody T46. ( D ) Gingipain cleavage sites in rtau-441 deduced from peptide fragments identified by MS for rtau-441 incubated with 1 or 10 nM gingipains. (a) T46 antibody epitope (red). (b) Tau-5 antibody epitope (red). (c) N-terminal tau fragment. (d) C-terminal tau fragment. (e) Kgp-generated tau fragments containing the VQIVYK sequence. (f) Kgp-generated fragments containing the VQIINK sequence. (g) An RgpB-generated tau fragment. *Cleavage sites identified at 1 nM gingipains.

    Techniques Used: Western Blot, Infection, Activity Assay, Negative Control, Incubation, Purification, Mass Spectrometry, Generated, Sequencing

    14) Product Images from "Sex Dimorphism in Periodontitis in Animal Models"

    Article Title: Sex Dimorphism in Periodontitis in Animal Models

    Journal: Journal of periodontal research

    doi: 10.1111/jre.12298

    Periodontal bone loss in P. gingivalis -infected female and male mice. The mm distance from the cementoenamel junction (CEJ) to the alveolar bone crest (ABC) was measured at 14 predetermined maxillary buccal sites and the readings were totaled for each
    Figure Legend Snippet: Periodontal bone loss in P. gingivalis -infected female and male mice. The mm distance from the cementoenamel junction (CEJ) to the alveolar bone crest (ABC) was measured at 14 predetermined maxillary buccal sites and the readings were totaled for each

    Techniques Used: Infection, Mouse Assay

    Relative expression of interested cytokines in the gingivae of P. gingivalis -infected female and male mice. Quantitative real-time PCR (qPCR) was used to determine gingival mRNA expression levels for the indicated molecules (normalized against GAPDH
    Figure Legend Snippet: Relative expression of interested cytokines in the gingivae of P. gingivalis -infected female and male mice. Quantitative real-time PCR (qPCR) was used to determine gingival mRNA expression levels for the indicated molecules (normalized against GAPDH

    Techniques Used: Expressing, Infection, Mouse Assay, Real-time Polymerase Chain Reaction

    15) Product Images from "Porphyromonas gulae Has Virulence and Immunological Characteristics Similar to Those of the Human Periodontal Pathogen Porphyromonas gingivalis"

    Article Title: Porphyromonas gulae Has Virulence and Immunological Characteristics Similar to Those of the Human Periodontal Pathogen Porphyromonas gingivalis

    Journal: Infection and Immunity

    doi: 10.1128/IAI.01500-15

    P. gulae is phagocytosed more readily than P. gingivalis. P. gingivalis (W50 and ATCC 33277) and P. gulae ATCC 51700 were labeled with pHrodo and examined for their ability to be phagocytosed by mouse macrophages (RAW 264.7) at increasing bacterium/cell
    Figure Legend Snippet: P. gulae is phagocytosed more readily than P. gingivalis. P. gingivalis (W50 and ATCC 33277) and P. gulae ATCC 51700 were labeled with pHrodo and examined for their ability to be phagocytosed by mouse macrophages (RAW 264.7) at increasing bacterium/cell

    Techniques Used: Labeling

    Macrophage cytokine response to P. gulae . RAW 264.7 macrophages were incubated with no bacteria (▩), P. gulae ATCC 51700 (□), P. gingivalis ATCC 33277 (), or P. gingivalis W50 (■) for 90 min before the cells were centrifuged and
    Figure Legend Snippet: Macrophage cytokine response to P. gulae . RAW 264.7 macrophages were incubated with no bacteria (▩), P. gulae ATCC 51700 (□), P. gingivalis ATCC 33277 (), or P. gingivalis W50 (■) for 90 min before the cells were centrifuged and

    Techniques Used: Incubation

    16) Product Images from "Porphyromonas gulae Has Virulence and Immunological Characteristics Similar to Those of the Human Periodontal Pathogen Porphyromonas gingivalis"

    Article Title: Porphyromonas gulae Has Virulence and Immunological Characteristics Similar to Those of the Human Periodontal Pathogen Porphyromonas gingivalis

    Journal: Infection and Immunity

    doi: 10.1128/IAI.01500-15

    P. gulae is phagocytosed more readily than P. gingivalis. P. gingivalis (W50 and ATCC 33277) and P. gulae ATCC 51700 were labeled with pHrodo and examined for their ability to be phagocytosed by mouse macrophages (RAW 264.7) at increasing bacterium/cell
    Figure Legend Snippet: P. gulae is phagocytosed more readily than P. gingivalis. P. gingivalis (W50 and ATCC 33277) and P. gulae ATCC 51700 were labeled with pHrodo and examined for their ability to be phagocytosed by mouse macrophages (RAW 264.7) at increasing bacterium/cell

    Techniques Used: Labeling

    Macrophage cytokine response to P. gulae . RAW 264.7 macrophages were incubated with no bacteria (▩), P. gulae ATCC 51700 (□), P. gingivalis ATCC 33277 (), or P. gingivalis W50 (■) for 90 min before the cells were centrifuged and
    Figure Legend Snippet: Macrophage cytokine response to P. gulae . RAW 264.7 macrophages were incubated with no bacteria (▩), P. gulae ATCC 51700 (□), P. gingivalis ATCC 33277 (), or P. gingivalis W50 (■) for 90 min before the cells were centrifuged and

    Techniques Used: Incubation

    17) Product Images from "Interferon Regulatory Factor 6 Promotes Keratinocyte Differentiation in Response to Porphyromonas gingivalis"

    Article Title: Interferon Regulatory Factor 6 Promotes Keratinocyte Differentiation in Response to Porphyromonas gingivalis

    Journal: Infection and Immunity

    doi: 10.1128/IAI.00858-16

    TLR2-independent stimulation of differentiation-associated gene expression in oral keratinocytes by P. gingivalis . OKF6 cells were transfected with a TLR2 (+) or control (–) siRNA (A to D) or with an IRAK1 (+) or control (–) siRNA (E to
    Figure Legend Snippet: TLR2-independent stimulation of differentiation-associated gene expression in oral keratinocytes by P. gingivalis . OKF6 cells were transfected with a TLR2 (+) or control (–) siRNA (A to D) or with an IRAK1 (+) or control (–) siRNA (E to

    Techniques Used: Expressing, Transfection

    RIPK4-dependent stimulation of differentiation-associated gene expression in oral keratinocytes by P. gingivalis . OKF6 cells were transfected with an RIPK4 (+) or control (–) siRNA. Thereafter, the cells were cultured with P. gingivalis (MOI of
    Figure Legend Snippet: RIPK4-dependent stimulation of differentiation-associated gene expression in oral keratinocytes by P. gingivalis . OKF6 cells were transfected with an RIPK4 (+) or control (–) siRNA. Thereafter, the cells were cultured with P. gingivalis (MOI of

    Techniques Used: Expressing, Transfection, Cell Culture

    Stimulation of RIPK4 expression in oral keratinocytes by P. gingivalis . (A) OKF6 cells were cultured with P. gingivalis (MOI of 100:1) for the times indicated, and RIPK4 mRNA levels were then measured ( n = 3). (B) OKF6 cells were cultured with P. gingivalis
    Figure Legend Snippet: Stimulation of RIPK4 expression in oral keratinocytes by P. gingivalis . (A) OKF6 cells were cultured with P. gingivalis (MOI of 100:1) for the times indicated, and RIPK4 mRNA levels were then measured ( n = 3). (B) OKF6 cells were cultured with P. gingivalis

    Techniques Used: Expressing, Cell Culture

    IRF6-dependent stimulation of differentiation-associated gene expression in oral keratinocytes by P. gingivalis . (A to D) OKF6 cells were cultured with P. gingivalis (MOI of 100:1) for the times indicated, and IVL (A), KRT13 (B), GRHL3 (C), and OVOL1
    Figure Legend Snippet: IRF6-dependent stimulation of differentiation-associated gene expression in oral keratinocytes by P. gingivalis . (A to D) OKF6 cells were cultured with P. gingivalis (MOI of 100:1) for the times indicated, and IVL (A), KRT13 (B), GRHL3 (C), and OVOL1

    Techniques Used: Expressing, Cell Culture

    IRF6-dependent stimulation of barrier function gene expression in oral keratinocytes by P. gingivalis . OKF6 cells were transfected with an IRF6 (+) or control (–) siRNA. Thereafter, the cells were cultured with P. gingivalis (MOI of 100:1) for
    Figure Legend Snippet: IRF6-dependent stimulation of barrier function gene expression in oral keratinocytes by P. gingivalis . OKF6 cells were transfected with an IRF6 (+) or control (–) siRNA. Thereafter, the cells were cultured with P. gingivalis (MOI of 100:1) for

    Techniques Used: Expressing, Transfection, Cell Culture

    18) Product Images from "Analysis of differential expression of tight junction proteins in cultured oral epithelial cells altered by Porphyromonas gingivalis, Porphyromonas gingivalis lipopolysaccharide, and extracellular adenosine triphosphate"

    Article Title: Analysis of differential expression of tight junction proteins in cultured oral epithelial cells altered by Porphyromonas gingivalis, Porphyromonas gingivalis lipopolysaccharide, and extracellular adenosine triphosphate

    Journal: International Journal of Oral Science

    doi: 10.1038/ijos.2017.51

    Tight junctions' staining. Occludin, JAM-A, claudin-1, claudin-4, claudin-15, and ZO-1 staining ( a – x ): H413 clone-1 cells treated with P. gingivalis infection ( a – f ), P. gingivalis LPS stimulation ( g – l ), and ATP plus P. gingivalis ( m – r ), or P. gingivalis LPS ( s – x ) for 2 h. Only moderate expression was observed for claudin-4 ( d ) after P. gingivalis infection; weak staining was detected for claudin-1 ( i ), and the majority of immune-reactivity was detected for claudin-15 ( k ) and ZO-1 ( l ) with P. gingivalis LPS treatment. After ATP pretreatment for 3 h, significant staining for occludin, JAM-A, claudin-1, claudin-4, claudin-15, and ZO-1 were observed in both groups ( m – x ), control (y), ATP-control (z), error bar, 20 μm. There were similar patterns with different treatments for 4 h (data not shown). ATP, adenosine triphosphate; JAM, junctional adhesion molecule; LPS, lipopolysaccharide; ZO, zonula occludens.
    Figure Legend Snippet: Tight junctions' staining. Occludin, JAM-A, claudin-1, claudin-4, claudin-15, and ZO-1 staining ( a – x ): H413 clone-1 cells treated with P. gingivalis infection ( a – f ), P. gingivalis LPS stimulation ( g – l ), and ATP plus P. gingivalis ( m – r ), or P. gingivalis LPS ( s – x ) for 2 h. Only moderate expression was observed for claudin-4 ( d ) after P. gingivalis infection; weak staining was detected for claudin-1 ( i ), and the majority of immune-reactivity was detected for claudin-15 ( k ) and ZO-1 ( l ) with P. gingivalis LPS treatment. After ATP pretreatment for 3 h, significant staining for occludin, JAM-A, claudin-1, claudin-4, claudin-15, and ZO-1 were observed in both groups ( m – x ), control (y), ATP-control (z), error bar, 20 μm. There were similar patterns with different treatments for 4 h (data not shown). ATP, adenosine triphosphate; JAM, junctional adhesion molecule; LPS, lipopolysaccharide; ZO, zonula occludens.

    Techniques Used: Staining, Infection, Expressing

    Tight junctions’ gene expressions. Gene expressions of occludin ( a ), JAM-A ( b ), claudin-1 ( c ), claudin-4 ( d ), claudin-15 ( e ), and ZO-1( f ). Significant changes in gene expressions altered by different treatments with P. gingivalis infection (2 and 4 h), P. gingivalis LPS stimulation (2 and 4 h), and ATP plus P. gingivalis or P. gingivalis LPS (2 and 4 h) in H413 clone-1 epithelial cells. * P
    Figure Legend Snippet: Tight junctions’ gene expressions. Gene expressions of occludin ( a ), JAM-A ( b ), claudin-1 ( c ), claudin-4 ( d ), claudin-15 ( e ), and ZO-1( f ). Significant changes in gene expressions altered by different treatments with P. gingivalis infection (2 and 4 h), P. gingivalis LPS stimulation (2 and 4 h), and ATP plus P. gingivalis or P. gingivalis LPS (2 and 4 h) in H413 clone-1 epithelial cells. * P

    Techniques Used: Infection

    Tight junctions' protein expressions. Western blots showing occludin ( a ), JAM-A ( b ), claudin-1 ( c ), claudin-4 ( d ), claudin-15 ( e ) protein bands corresponding to their gene expression in H413 clone-1 epithelial cells in response to P. gingivalis , P. gingivalis LPS, and ATP plus P. gingivalis or P. gingivalis LPS. ATP, adenosine triphosphate; JAM, junctional adhesion molecule; LPS, lipopolysaccharide.
    Figure Legend Snippet: Tight junctions' protein expressions. Western blots showing occludin ( a ), JAM-A ( b ), claudin-1 ( c ), claudin-4 ( d ), claudin-15 ( e ) protein bands corresponding to their gene expression in H413 clone-1 epithelial cells in response to P. gingivalis , P. gingivalis LPS, and ATP plus P. gingivalis or P. gingivalis LPS. ATP, adenosine triphosphate; JAM, junctional adhesion molecule; LPS, lipopolysaccharide.

    Techniques Used: Western Blot, Expressing

    19) Product Images from "Regulation of the Peptidoglycan Amidase PGLYRP2 in Epithelial Cells by Interleukin-36γ"

    Article Title: Regulation of the Peptidoglycan Amidase PGLYRP2 in Epithelial Cells by Interleukin-36γ

    Journal: Infection and Immunity

    doi: 10.1128/IAI.00384-18

    Stimulation of PGLYRP2 and hBD2 expression in oral epithelial cells by bacteria. (A and B) TIGK cells were cultured with wild-type P. gingivalis or P. gingivalis KDP136 (Kgp/Rgp-null mutant) at an MOI of 100:1 for the times indicated. PGLYRP2 (A) and hBD2 (B) mRNA levels were then measured ( n = 3). (C to H) TIGK cells were cultured with F. nucleatum (C and F), S. gordonii (D and G), and S. sanguinis (E and H) at an MOI of 100:1 for the times indicated. PGLYRP2 (C to E) and hBD2 (F to H) mRNA levels were then measured ( n = 3). ***, P
    Figure Legend Snippet: Stimulation of PGLYRP2 and hBD2 expression in oral epithelial cells by bacteria. (A and B) TIGK cells were cultured with wild-type P. gingivalis or P. gingivalis KDP136 (Kgp/Rgp-null mutant) at an MOI of 100:1 for the times indicated. PGLYRP2 (A) and hBD2 (B) mRNA levels were then measured ( n = 3). (C to H) TIGK cells were cultured with F. nucleatum (C and F), S. gordonii (D and G), and S. sanguinis (E and H) at an MOI of 100:1 for the times indicated. PGLYRP2 (C to E) and hBD2 (F to H) mRNA levels were then measured ( n = 3). ***, P

    Techniques Used: Expressing, Cell Culture, Mutagenesis

    20) Product Images from "Detection of hydrogen cyanide from oral anaerobes by cavity ring down spectroscopy"

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

    Journal: Scientific Reports

    doi: 10.1038/srep22577

    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
    Figure Legend 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

    Techniques Used:

    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.
    Figure Legend 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.

    Techniques Used:

    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.
    Figure Legend 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.

    Techniques Used:

    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
    Figure Legend 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

    Techniques Used: Concentration Assay

    21) Product Images from "Impact of aging on TREM-1 responses in the periodontium: a cross-sectional study in an elderly population"

    Article Title: Impact of aging on TREM-1 responses in the periodontium: a cross-sectional study in an elderly population

    Journal: BMC Infectious Diseases

    doi: 10.1186/s12879-016-1778-6

    Oral microbiota levels in the subgingival plaque from elderly individuals with periodontal health ( n = 17), gingivitis ( n = 19), and chronic periodontitis ( n = 15). The individual values represent bacterial counts per sample in the subgingival plaque ( a ) Total bacterial counts, ( b ) F. nucleatum , ( c ) P. intermedia , ( d ) P. gingivalis , ( e ) T. denticola , and ( f ) T. forsythia . The horizontal lines in the middle of the box represent the median and whiskers are drawn down to the 10 th through 90 th percentiles. The points below and above the whiskers are drawn as individual dots
    Figure Legend Snippet: Oral microbiota levels in the subgingival plaque from elderly individuals with periodontal health ( n = 17), gingivitis ( n = 19), and chronic periodontitis ( n = 15). The individual values represent bacterial counts per sample in the subgingival plaque ( a ) Total bacterial counts, ( b ) F. nucleatum , ( c ) P. intermedia , ( d ) P. gingivalis , ( e ) T. denticola , and ( f ) T. forsythia . The horizontal lines in the middle of the box represent the median and whiskers are drawn down to the 10 th through 90 th percentiles. The points below and above the whiskers are drawn as individual dots

    Techniques Used:

    22) Product Images from "Porphyromonas gingivalis can invade periodontal ligament stem cells"

    Article Title: Porphyromonas gingivalis can invade periodontal ligament stem cells

    Journal: BMC Microbiology

    doi: 10.1186/s12866-017-0950-5

    The efficiency of P. gingivalis ATCC 33277 infection in PDLSCs. The infection rate of P. gingivalis in PDLSCs at MOIs of 50, 100, 200, and 500 were 5.83%, 8.12%, 7.77% and 7.53%, respectively, according to the agar plate culture method. The efficiencies of P. gingivalis infection of PDLSCs at MOIs of 50, 100, 200, and 500 were 6.74%, 10.56%, 10.36% and 9.78%, respectively, according to the q-PCR method. The data are presented as the mean ± SD of triplicate experiments. *Significant difference ( P
    Figure Legend Snippet: The efficiency of P. gingivalis ATCC 33277 infection in PDLSCs. The infection rate of P. gingivalis in PDLSCs at MOIs of 50, 100, 200, and 500 were 5.83%, 8.12%, 7.77% and 7.53%, respectively, according to the agar plate culture method. The efficiencies of P. gingivalis infection of PDLSCs at MOIs of 50, 100, 200, and 500 were 6.74%, 10.56%, 10.36% and 9.78%, respectively, according to the q-PCR method. The data are presented as the mean ± SD of triplicate experiments. *Significant difference ( P

    Techniques Used: Infection, Polymerase Chain Reaction

    PDLSCs infected with P. gingivalis ATCC 33277 under transmission electron microscopy. The nucleus was large and round, and organelles were abundant in PDLSCs ( a ). P. gingivalis ATCC 33277 could invade PDLSCs after 2 h of incubation ( b , c - A ). Endocytic vacuoles were not found surrounding internalized P. gingivalis . The bumps observed were stretched membrane where the PDLSCs packaged P. gingivalis ATCC 33277 ( b , c - B )
    Figure Legend Snippet: PDLSCs infected with P. gingivalis ATCC 33277 under transmission electron microscopy. The nucleus was large and round, and organelles were abundant in PDLSCs ( a ). P. gingivalis ATCC 33277 could invade PDLSCs after 2 h of incubation ( b , c - A ). Endocytic vacuoles were not found surrounding internalized P. gingivalis . The bumps observed were stretched membrane where the PDLSCs packaged P. gingivalis ATCC 33277 ( b , c - B )

    Techniques Used: Infection, Transmission Assay, Electron Microscopy, Incubation

    23) Product Images from "Antibacterial activity and cytocompatibility of an implant coating consisting of TiO2 nanotubes combined with a GL13K antimicrobial peptide"

    Article Title: Antibacterial activity and cytocompatibility of an implant coating consisting of TiO2 nanotubes combined with a GL13K antimicrobial peptide

    Journal: International Journal of Nanomedicine

    doi: 10.2147/IJN.S128775

    Evaluation of antimicrobial activity of the MNA-TNTs, GL13K-TNTs, and TNTs against ( A ) Porphyromonas gingivalis ATCC 33277 and ( B ) Fusobacterium nucleatum ATCC 25586, assessed using disk-diffusion assay. Abbreviations: MNA, metronidazole; MNA-TNTs, MNA-immobilized TNTs; TNTs, TiO 2 nanotubes; ATCC, American Type Culture Collection.
    Figure Legend Snippet: Evaluation of antimicrobial activity of the MNA-TNTs, GL13K-TNTs, and TNTs against ( A ) Porphyromonas gingivalis ATCC 33277 and ( B ) Fusobacterium nucleatum ATCC 25586, assessed using disk-diffusion assay. Abbreviations: MNA, metronidazole; MNA-TNTs, MNA-immobilized TNTs; TNTs, TiO 2 nanotubes; ATCC, American Type Culture Collection.

    Techniques Used: Activity Assay, Diffusion-based Assay

    24) Product Images from "Antibacterial Effect of Curcumin against Clinically Isolated Porphyromonas gingivalis and Connective Tissue Reactions to Curcumin Gel in the Subcutaneous Tissue of Rats"

    Article Title: Antibacterial Effect of Curcumin against Clinically Isolated Porphyromonas gingivalis and Connective Tissue Reactions to Curcumin Gel in the Subcutaneous Tissue of Rats

    Journal: BioMed Research International

    doi: 10.1155/2019/6810936

    16S rDNA gene sequence of the purified colonies of P. gingivalis done in Macrogen, South Korea.
    Figure Legend Snippet: 16S rDNA gene sequence of the purified colonies of P. gingivalis done in Macrogen, South Korea.

    Techniques Used: Sequencing, Purification

    Pure isolated bacterial colonies of P. gingivalis after 7 (a) and 10 (b) days on Columbia agar appearing as round, smooth, shiny, and convex black colonies. (c) Gram-negative short rods of P. gingivalis and (d) agar formed media cultured from the tube with the MIC of curcumin showing no P. gingivalis growth.
    Figure Legend Snippet: Pure isolated bacterial colonies of P. gingivalis after 7 (a) and 10 (b) days on Columbia agar appearing as round, smooth, shiny, and convex black colonies. (c) Gram-negative short rods of P. gingivalis and (d) agar formed media cultured from the tube with the MIC of curcumin showing no P. gingivalis growth.

    Techniques Used: Isolation, Cell Culture

    25) Product Images from "Er:YAG Laser Irradiation Reduces Microbial Viability When Used in Combination with Irrigation with Sodium Hypochlorite, Chlorhexidine, and Hydrogen Peroxide"

    Article Title: Er:YAG Laser Irradiation Reduces Microbial Viability When Used in Combination with Irrigation with Sodium Hypochlorite, Chlorhexidine, and Hydrogen Peroxide

    Journal: Microorganisms

    doi: 10.3390/microorganisms7120612

    Effectiveness of erbium-doped yttrium aluminum garnet (Er:YAG) laser treatment with or without irrigation on the viability of P. gingivalis. Bacterial strains were grown anaerobically in brain heart infusion (BHI) broth. For treatment, cultures were aliquoted onto a 96-well plate and treated with laser irradiation, anti-microbial at listed concentration, or combination therapy. Then, all samples were plated on trypticase soy agar, TSA blood agar plates post-treatment. After incubation, colonies were counted to judge the efficiency of treatment. Laser-treated samples were treated with an Er:YAG laser at the following settings: 40 mJ; 40 Hz; 1.6 W, 20 seconds, 300 µs short pulse duration, contact mode. Untreated cultures served as controls. Data is representative of four biological replicates. ( p
    Figure Legend Snippet: Effectiveness of erbium-doped yttrium aluminum garnet (Er:YAG) laser treatment with or without irrigation on the viability of P. gingivalis. Bacterial strains were grown anaerobically in brain heart infusion (BHI) broth. For treatment, cultures were aliquoted onto a 96-well plate and treated with laser irradiation, anti-microbial at listed concentration, or combination therapy. Then, all samples were plated on trypticase soy agar, TSA blood agar plates post-treatment. After incubation, colonies were counted to judge the efficiency of treatment. Laser-treated samples were treated with an Er:YAG laser at the following settings: 40 mJ; 40 Hz; 1.6 W, 20 seconds, 300 µs short pulse duration, contact mode. Untreated cultures served as controls. Data is representative of four biological replicates. ( p

    Techniques Used: Irradiation, Concentration Assay, Incubation

    26) Product Images from "Interferon Regulatory Factor 6 Promotes Keratinocyte Differentiation in Response to Porphyromonas gingivalis"

    Article Title: Interferon Regulatory Factor 6 Promotes Keratinocyte Differentiation in Response to Porphyromonas gingivalis

    Journal: Infection and Immunity

    doi: 10.1128/IAI.00858-16

    TLR2-independent stimulation of differentiation-associated gene expression in oral keratinocytes by P. gingivalis . OKF6 cells were transfected with a TLR2 (+) or control (–) siRNA (A to D) or with an IRAK1 (+) or control (–) siRNA (E to
    Figure Legend Snippet: TLR2-independent stimulation of differentiation-associated gene expression in oral keratinocytes by P. gingivalis . OKF6 cells were transfected with a TLR2 (+) or control (–) siRNA (A to D) or with an IRAK1 (+) or control (–) siRNA (E to

    Techniques Used: Expressing, Transfection

    RIPK4-dependent stimulation of differentiation-associated gene expression in oral keratinocytes by P. gingivalis . OKF6 cells were transfected with an RIPK4 (+) or control (–) siRNA. Thereafter, the cells were cultured with P. gingivalis (MOI of
    Figure Legend Snippet: RIPK4-dependent stimulation of differentiation-associated gene expression in oral keratinocytes by P. gingivalis . OKF6 cells were transfected with an RIPK4 (+) or control (–) siRNA. Thereafter, the cells were cultured with P. gingivalis (MOI of

    Techniques Used: Expressing, Transfection, Cell Culture

    Stimulation of RIPK4 expression in oral keratinocytes by P. gingivalis . (A) OKF6 cells were cultured with P. gingivalis (MOI of 100:1) for the times indicated, and RIPK4 mRNA levels were then measured ( n = 3). (B) OKF6 cells were cultured with P. gingivalis
    Figure Legend Snippet: Stimulation of RIPK4 expression in oral keratinocytes by P. gingivalis . (A) OKF6 cells were cultured with P. gingivalis (MOI of 100:1) for the times indicated, and RIPK4 mRNA levels were then measured ( n = 3). (B) OKF6 cells were cultured with P. gingivalis

    Techniques Used: Expressing, Cell Culture

    IRF6-dependent stimulation of differentiation-associated gene expression in oral keratinocytes by P. gingivalis . (A to D) OKF6 cells were cultured with P. gingivalis (MOI of 100:1) for the times indicated, and IVL (A), KRT13 (B), GRHL3 (C), and OVOL1
    Figure Legend Snippet: IRF6-dependent stimulation of differentiation-associated gene expression in oral keratinocytes by P. gingivalis . (A to D) OKF6 cells were cultured with P. gingivalis (MOI of 100:1) for the times indicated, and IVL (A), KRT13 (B), GRHL3 (C), and OVOL1

    Techniques Used: Expressing, Cell Culture

    IRF6-dependent stimulation of barrier function gene expression in oral keratinocytes by P. gingivalis . OKF6 cells were transfected with an IRF6 (+) or control (–) siRNA. Thereafter, the cells were cultured with P. gingivalis (MOI of 100:1) for
    Figure Legend Snippet: IRF6-dependent stimulation of barrier function gene expression in oral keratinocytes by P. gingivalis . OKF6 cells were transfected with an IRF6 (+) or control (–) siRNA. Thereafter, the cells were cultured with P. gingivalis (MOI of 100:1) for

    Techniques Used: Expressing, Transfection, Cell Culture

    27) Product Images from "Identification of the Binding Domain of Streptococcus oralis Glyceraldehyde-3-Phosphate Dehydrogenase for Porphyromonas gingivalis Major Fimbriae ▿"

    Article Title: Identification of the Binding Domain of Streptococcus oralis Glyceraldehyde-3-Phosphate Dehydrogenase for Porphyromonas gingivalis Major Fimbriae ▿

    Journal: Infection and Immunity

    doi: 10.1128/IAI.00439-09

    Inhibitory effects of pep166-183 on biofilm formation by P. gingivalis ATCC 33277 with various streptococci as determined by CLSM. HI-stained streptococci (5 × 10 7 CFU/well; red) were inoculated into individual chambers coated with human whole
    Figure Legend Snippet: Inhibitory effects of pep166-183 on biofilm formation by P. gingivalis ATCC 33277 with various streptococci as determined by CLSM. HI-stained streptococci (5 × 10 7 CFU/well; red) were inoculated into individual chambers coated with human whole

    Techniques Used: Confocal Laser Scanning Microscopy, Staining

    Inhibitory effects of pep166-183 on biofilm formation by S. oralis ATCC 9811 and P. gingivalis strains with different types of fimbriae as determined by CLSM. HI-stained S. oralis ATCC 9811 (5 × 10 7 CFU/well; red) was inoculated into individual
    Figure Legend Snippet: Inhibitory effects of pep166-183 on biofilm formation by S. oralis ATCC 9811 and P. gingivalis strains with different types of fimbriae as determined by CLSM. HI-stained S. oralis ATCC 9811 (5 × 10 7 CFU/well; red) was inoculated into individual

    Techniques Used: Confocal Laser Scanning Microscopy, Staining

    28) Product Images from "Amniotic fluid and maternal race influence responsiveness of fetal membranes to bacteria ☆"

    Article Title: Amniotic fluid and maternal race influence responsiveness of fetal membranes to bacteria ☆

    Journal: Journal of reproductive immunology

    doi: 10.1016/j.jri.2012.07.006

    Racial disparity in the effect of amniotic fluid (AF) on IL-1β (A) and TNF-α (B) production by fetal membranes. Shown on a log-scale are least-squares means ± SEM for membrane cultures stimulated with medium alone (CTL), E. coli (EC), Group B streptococci (GBS), G. vaginalis (GV), lipopolysaccharide (LPS), M. hominis (MH), P. gingivalis (PG), U. parvum (UP) or U. urealyticum (UU) stratified by race. Pairs of bars marked with asterisks indicate statistically significant effects of AF at * P ≤ 0.05 or ** P ≤ 0.01 for that race and bacterial stimulation.
    Figure Legend Snippet: Racial disparity in the effect of amniotic fluid (AF) on IL-1β (A) and TNF-α (B) production by fetal membranes. Shown on a log-scale are least-squares means ± SEM for membrane cultures stimulated with medium alone (CTL), E. coli (EC), Group B streptococci (GBS), G. vaginalis (GV), lipopolysaccharide (LPS), M. hominis (MH), P. gingivalis (PG), U. parvum (UP) or U. urealyticum (UU) stratified by race. Pairs of bars marked with asterisks indicate statistically significant effects of AF at * P ≤ 0.05 or ** P ≤ 0.01 for that race and bacterial stimulation.

    Techniques Used: CTL Assay

    Racial disparity for the effect of amniotic fluid (AF) on IL-10 (A) and IL-8 (B) production by fetal membranes. Shown are least-squares means ± SEM for membrane cultures stimulated with medium alone (CTL), E. coli (EC), Group B streptococci (GBS), G. vaginalis (GV), lipopolysaccharide (LPS), M. hominis (MH), P. gingivalis (PG), U. parvum (UP) or U. urealyticum (UU) stratified by race. Bars marked with asterisks indicate statistically significant effects of AF at * P 0.05 ≤ or ** P ≤ 0.01 for that race and bacterial stimulation.
    Figure Legend Snippet: Racial disparity for the effect of amniotic fluid (AF) on IL-10 (A) and IL-8 (B) production by fetal membranes. Shown are least-squares means ± SEM for membrane cultures stimulated with medium alone (CTL), E. coli (EC), Group B streptococci (GBS), G. vaginalis (GV), lipopolysaccharide (LPS), M. hominis (MH), P. gingivalis (PG), U. parvum (UP) or U. urealyticum (UU) stratified by race. Bars marked with asterisks indicate statistically significant effects of AF at * P 0.05 ≤ or ** P ≤ 0.01 for that race and bacterial stimulation.

    Techniques Used: CTL Assay

    Racial disparity in the effect of amniotic fluid on IL-1b (A) and TNF-α (B) production by fetal membranes. Shown are least-squares means ± SEM for membrane cultures stimulated with medium alone (CTL), or cultures stimulated with E. coli (EC), Group B Streptococci (GBS), G. vaginalis (GV)-, Lippopolysaccharide (LPS), M. hominis (MH), P. gingivalis (PG), U. parvum (UP) or U. urealyaticum (UU) stratified by race. Pairs of bars marked with asterisks indicate statistically significant effects of amniotic fluid at * P
    Figure Legend Snippet: Racial disparity in the effect of amniotic fluid on IL-1b (A) and TNF-α (B) production by fetal membranes. Shown are least-squares means ± SEM for membrane cultures stimulated with medium alone (CTL), or cultures stimulated with E. coli (EC), Group B Streptococci (GBS), G. vaginalis (GV)-, Lippopolysaccharide (LPS), M. hominis (MH), P. gingivalis (PG), U. parvum (UP) or U. urealyaticum (UU) stratified by race. Pairs of bars marked with asterisks indicate statistically significant effects of amniotic fluid at * P

    Techniques Used: CTL Assay

    29) Product Images from "Biological and immunotoxicity evaluation of antimicrobial peptide-loaded coatings using a layer-by-layer process on titanium"

    Article Title: Biological and immunotoxicity evaluation of antimicrobial peptide-loaded coatings using a layer-by-layer process on titanium

    Journal: Scientific Reports

    doi: 10.1038/srep16336

    Sustained antimicrobial activity showed that P. gingivalis ( A ) and S. aureus ( B ) efficiently inhibited by CS-(HA-AMPCol)10 samples for up to one month. Samples were collected by two methods, collecting part of the broth without refilling (method 1), and gathering all the broth then adding an equal amount of broth (method 2).
    Figure Legend Snippet: Sustained antimicrobial activity showed that P. gingivalis ( A ) and S. aureus ( B ) efficiently inhibited by CS-(HA-AMPCol)10 samples for up to one month. Samples were collected by two methods, collecting part of the broth without refilling (method 1), and gathering all the broth then adding an equal amount of broth (method 2).

    Techniques Used: Activity Assay

    30) Product Images from "Egg yolk immunoglobulin interactions with Porphyromonas gingivalis to impact periodontal inflammation and halitosis"

    Article Title: Egg yolk immunoglobulin interactions with Porphyromonas gingivalis to impact periodontal inflammation and halitosis

    Journal: AMB Express

    doi: 10.1186/s13568-018-0706-0

    Effect of IgY on rat gingivitis and bad breath induced by P. gingivalis . The scores for oral health index are shown as mean ± SD (n = 3). *Means different at p
    Figure Legend Snippet: Effect of IgY on rat gingivitis and bad breath induced by P. gingivalis . The scores for oral health index are shown as mean ± SD (n = 3). *Means different at p

    Techniques Used:

    The inhibitory effect of IgY on the growth of P. gingivalis . P. gingivalis was cultured in artificial saliva medium containing 0 (control), 0.0037 (low-dose), 7.4 (mid-dose) or 370 (high-dose) mmol/l of IgY. Data were obtained using three independent experiments with three measurements in each experiment, and are shown as mean ± SD (n = 3). *Means different at p
    Figure Legend Snippet: The inhibitory effect of IgY on the growth of P. gingivalis . P. gingivalis was cultured in artificial saliva medium containing 0 (control), 0.0037 (low-dose), 7.4 (mid-dose) or 370 (high-dose) mmol/l of IgY. Data were obtained using three independent experiments with three measurements in each experiment, and are shown as mean ± SD (n = 3). *Means different at p

    Techniques Used: Cell Culture

    31) Product Images from "Epithelial Cell Surface Sites Involved in the Polyvalent Adherence of Porphyromonas gingivalis: a Convincing Role for Neuraminic Acid and Glucuronic Acid "

    Article Title: Epithelial Cell Surface Sites Involved in the Polyvalent Adherence of Porphyromonas gingivalis: a Convincing Role for Neuraminic Acid and Glucuronic Acid

    Journal: Infection and Immunity

    doi: 10.1128/IAI.71.2.991-996.2003

    Flow cytometry histograms showing inhibition of adherence of P. gingivalis to KB cells. FITC-labeled P. gingivalis (100 bacteria per cell) were allowed to bind to suspended KB cells (2 × 10 5 ). (A) The eukaryotic cells were preincubated for 45 min at 4°C with anti-keratinocyte immunserum (red line) or nonimmune serum (blue line) at 1:5 dilution. (B) The FITC-labeled P. gingivalis were preincubated for 1 h at room temperature with anti- P. gingivalis antiserum (red line) or nonimmune serum (blue line) at 1:5 dilution. In both figures, cell autofluorescence is visualized in green.
    Figure Legend Snippet: Flow cytometry histograms showing inhibition of adherence of P. gingivalis to KB cells. FITC-labeled P. gingivalis (100 bacteria per cell) were allowed to bind to suspended KB cells (2 × 10 5 ). (A) The eukaryotic cells were preincubated for 45 min at 4°C with anti-keratinocyte immunserum (red line) or nonimmune serum (blue line) at 1:5 dilution. (B) The FITC-labeled P. gingivalis were preincubated for 1 h at room temperature with anti- P. gingivalis antiserum (red line) or nonimmune serum (blue line) at 1:5 dilution. In both figures, cell autofluorescence is visualized in green.

    Techniques Used: Flow Cytometry, Cytometry, Inhibition, Labeling

    Confocal photomicrographs. KB cells were incubated with FITC-labeled P. gingivalis at an infection multiplicity of 100 for 30 min at room temperature, in the presence of 25 mM glucuronate (A) or not (B).
    Figure Legend Snippet: Confocal photomicrographs. KB cells were incubated with FITC-labeled P. gingivalis at an infection multiplicity of 100 for 30 min at room temperature, in the presence of 25 mM glucuronate (A) or not (B).

    Techniques Used: Incubation, Labeling, Infection

    32) Product Images from "Toll-Like Receptor 9-Mediated Inflammation Triggers Alveolar Bone Loss in Experimental Murine Periodontitis"

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

    Journal: Infection and Immunity

    doi: 10.1128/IAI.00424-15

    TLR9 KO (TLR9 −/− ) mice are resistant to P. gingivalis -instigated periodontal bone loss. Groups of mice (WT [ n = 17] and TLR9 −/− [ n = 47]) were infected with P. gingivalis (Pg) or sham infected and euthanized 42 days later. Measurements were performed in defleshed maxillae. The data are represented as the mean results ± SD ( n = 64 mice). (A) Distance (in millimeters) between the cemento-enamel junction (CEJ) and alveolar bone crest (ABC) in each group of animals. (B) Amount of bone change in WT and TLR9 −/− mice. Negative values indicate bone loss in P. gingivalis -inoculated mice relative to the results for vehicle-inoculated (sham) controls. (C) Representative micro-CT images of maxillae from each group of mice. *, P
    Figure Legend Snippet: TLR9 KO (TLR9 −/− ) mice are resistant to P. gingivalis -instigated periodontal bone loss. Groups of mice (WT [ n = 17] and TLR9 −/− [ n = 47]) were infected with P. gingivalis (Pg) or sham infected and euthanized 42 days later. Measurements were performed in defleshed maxillae. The data are represented as the mean results ± SD ( n = 64 mice). (A) Distance (in millimeters) between the cemento-enamel junction (CEJ) and alveolar bone crest (ABC) in each group of animals. (B) Amount of bone change in WT and TLR9 −/− mice. Negative values indicate bone loss in P. gingivalis -inoculated mice relative to the results for vehicle-inoculated (sham) controls. (C) Representative micro-CT images of maxillae from each group of mice. *, P

    Techniques Used: Mouse Assay, Infection, Micro-CT

    Inflammatory molecule expression in gingival tissues. WT and TLR9 −/− mice were orally inoculated with P. gingivalis or vehicle only (sham) and euthanized 42 days later. Gingival tissues around maxillary molars were excised and processed for qPCR analyses to determine mRNA expression of TNF (A), IL-6 (B), and RANKL (C). Results are reported as fold induction after normalization to GAPDH. The data shown are the mean results ± SD ( n = 5 or 6 mice per group) and were analyzed using the unpaired t test. *, P
    Figure Legend Snippet: Inflammatory molecule expression in gingival tissues. WT and TLR9 −/− mice were orally inoculated with P. gingivalis or vehicle only (sham) and euthanized 42 days later. Gingival tissues around maxillary molars were excised and processed for qPCR analyses to determine mRNA expression of TNF (A), IL-6 (B), and RANKL (C). Results are reported as fold induction after normalization to GAPDH. The data shown are the mean results ± SD ( n = 5 or 6 mice per group) and were analyzed using the unpaired t test. *, P

    Techniques Used: Expressing, Mouse Assay, Real-time Polymerase Chain Reaction

    Comparison of proinflammatory cytokine production in WT versus TLR9 −/− macrophages (A, B) and splenocytes (C, D) in response to P. gingivalis challenge after 24 h. The cells were stimulated with heat-killed P. gingivalis (MOI of 1:100), P. gingivalis DNA (100 ng/μl), and ODN 1668 (TLR9 agonist), and cell-free supernatants were analyzed for the presence of IL-6 and TNF using ELISA. Comparisons between the results for WT and TLR9 KO cells were performed using the unpaired student t test. The levels of IL-6 and TNF production were significantly reduced in TLR9 −/− macrophages (A, B) and splenocytes (C, D) compared to the levels in WT cells. The results shown are representative of at least 3 independent experiments that were run in triplicates. The data shown are the mean results ± SD ( n = 9). *, P
    Figure Legend Snippet: Comparison of proinflammatory cytokine production in WT versus TLR9 −/− macrophages (A, B) and splenocytes (C, D) in response to P. gingivalis challenge after 24 h. The cells were stimulated with heat-killed P. gingivalis (MOI of 1:100), P. gingivalis DNA (100 ng/μl), and ODN 1668 (TLR9 agonist), and cell-free supernatants were analyzed for the presence of IL-6 and TNF using ELISA. Comparisons between the results for WT and TLR9 KO cells were performed using the unpaired student t test. The levels of IL-6 and TNF production were significantly reduced in TLR9 −/− macrophages (A, B) and splenocytes (C, D) compared to the levels in WT cells. The results shown are representative of at least 3 independent experiments that were run in triplicates. The data shown are the mean results ± SD ( n = 9). *, P

    Techniques Used: Enzyme-linked Immunosorbent Assay

    Comparison of P. gingivalis levels within the periodontal tissues (A) and P. gingivalis -specific antibody responses in serum (B) in TLR9 −/− and WT mice. Each strain was orally inoculated with P. gingivalis or vehicle only (sham). The P. gingivalis levels were determined by qPCR of the ISPg1 gene ( P. gingivalis ) at 2 weeks postinfection. There was no statistically significant difference in P. gingivalis levels among groups. P. gingivalis -specific antibody responses were determined at the termination of the experiment. The antibody titers in TLR9 −/− mice and WT mice infected with P. gingivalis were significantly higher than the titers in the uninfected mice ( P
    Figure Legend Snippet: Comparison of P. gingivalis levels within the periodontal tissues (A) and P. gingivalis -specific antibody responses in serum (B) in TLR9 −/− and WT mice. Each strain was orally inoculated with P. gingivalis or vehicle only (sham). The P. gingivalis levels were determined by qPCR of the ISPg1 gene ( P. gingivalis ) at 2 weeks postinfection. There was no statistically significant difference in P. gingivalis levels among groups. P. gingivalis -specific antibody responses were determined at the termination of the experiment. The antibody titers in TLR9 −/− mice and WT mice infected with P. gingivalis were significantly higher than the titers in the uninfected mice ( P

    Techniques Used: Mouse Assay, Real-time Polymerase Chain Reaction, Infection

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

    Techniques Used: Enzyme-linked Immunosorbent Assay

    33) Product Images from "Toll-Like Receptor 9-Mediated Inflammation Triggers Alveolar Bone Loss in Experimental Murine Periodontitis"

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

    Journal: Infection and Immunity

    doi: 10.1128/IAI.00424-15

    TLR9 KO (TLR9 −/− ) mice are resistant to P. gingivalis -instigated periodontal bone loss. Groups of mice (WT [ n = 17] and TLR9 −/− [ n = 47]) were infected with P. gingivalis (Pg) or sham infected and euthanized 42 days later. Measurements were performed in defleshed maxillae. The data are represented as the mean results ± SD ( n = 64 mice). (A) Distance (in millimeters) between the cemento-enamel junction (CEJ) and alveolar bone crest (ABC) in each group of animals. (B) Amount of bone change in WT and TLR9 −/− mice. Negative values indicate bone loss in P. gingivalis -inoculated mice relative to the results for vehicle-inoculated (sham) controls. (C) Representative micro-CT images of maxillae from each group of mice. *, P
    Figure Legend Snippet: TLR9 KO (TLR9 −/− ) mice are resistant to P. gingivalis -instigated periodontal bone loss. Groups of mice (WT [ n = 17] and TLR9 −/− [ n = 47]) were infected with P. gingivalis (Pg) or sham infected and euthanized 42 days later. Measurements were performed in defleshed maxillae. The data are represented as the mean results ± SD ( n = 64 mice). (A) Distance (in millimeters) between the cemento-enamel junction (CEJ) and alveolar bone crest (ABC) in each group of animals. (B) Amount of bone change in WT and TLR9 −/− mice. Negative values indicate bone loss in P. gingivalis -inoculated mice relative to the results for vehicle-inoculated (sham) controls. (C) Representative micro-CT images of maxillae from each group of mice. *, P

    Techniques Used: Mouse Assay, Infection, Micro-CT

    Inflammatory molecule expression in gingival tissues. WT and TLR9 −/− mice were orally inoculated with P. gingivalis or vehicle only (sham) and euthanized 42 days later. Gingival tissues around maxillary molars were excised and processed for qPCR analyses to determine mRNA expression of TNF (A), IL-6 (B), and RANKL (C). Results are reported as fold induction after normalization to GAPDH. The data shown are the mean results ± SD ( n = 5 or 6 mice per group) and were analyzed using the unpaired t test. *, P
    Figure Legend Snippet: Inflammatory molecule expression in gingival tissues. WT and TLR9 −/− mice were orally inoculated with P. gingivalis or vehicle only (sham) and euthanized 42 days later. Gingival tissues around maxillary molars were excised and processed for qPCR analyses to determine mRNA expression of TNF (A), IL-6 (B), and RANKL (C). Results are reported as fold induction after normalization to GAPDH. The data shown are the mean results ± SD ( n = 5 or 6 mice per group) and were analyzed using the unpaired t test. *, P

    Techniques Used: Expressing, Mouse Assay, Real-time Polymerase Chain Reaction

    Comparison of proinflammatory cytokine production in WT versus TLR9 −/− macrophages (A, B) and splenocytes (C, D) in response to P. gingivalis challenge after 24 h. The cells were stimulated with heat-killed P. gingivalis (MOI of 1:100), P. gingivalis DNA (100 ng/μl), and ODN 1668 (TLR9 agonist), and cell-free supernatants were analyzed for the presence of IL-6 and TNF using ELISA. Comparisons between the results for WT and TLR9 KO cells were performed using the unpaired student t test. The levels of IL-6 and TNF production were significantly reduced in TLR9 −/− macrophages (A, B) and splenocytes (C, D) compared to the levels in WT cells. The results shown are representative of at least 3 independent experiments that were run in triplicates. The data shown are the mean results ± SD ( n = 9). *, P
    Figure Legend Snippet: Comparison of proinflammatory cytokine production in WT versus TLR9 −/− macrophages (A, B) and splenocytes (C, D) in response to P. gingivalis challenge after 24 h. The cells were stimulated with heat-killed P. gingivalis (MOI of 1:100), P. gingivalis DNA (100 ng/μl), and ODN 1668 (TLR9 agonist), and cell-free supernatants were analyzed for the presence of IL-6 and TNF using ELISA. Comparisons between the results for WT and TLR9 KO cells were performed using the unpaired student t test. The levels of IL-6 and TNF production were significantly reduced in TLR9 −/− macrophages (A, B) and splenocytes (C, D) compared to the levels in WT cells. The results shown are representative of at least 3 independent experiments that were run in triplicates. The data shown are the mean results ± SD ( n = 9). *, P

    Techniques Used: Enzyme-linked Immunosorbent Assay

    Comparison of P. gingivalis levels within the periodontal tissues (A) and P. gingivalis -specific antibody responses in serum (B) in TLR9 −/− and WT mice. Each strain was orally inoculated with P. gingivalis or vehicle only (sham). The P. gingivalis levels were determined by qPCR of the ISPg1 gene ( P. gingivalis ) at 2 weeks postinfection. There was no statistically significant difference in P. gingivalis levels among groups. P. gingivalis -specific antibody responses were determined at the termination of the experiment. The antibody titers in TLR9 −/− mice and WT mice infected with P. gingivalis were significantly higher than the titers in the uninfected mice ( P
    Figure Legend Snippet: Comparison of P. gingivalis levels within the periodontal tissues (A) and P. gingivalis -specific antibody responses in serum (B) in TLR9 −/− and WT mice. Each strain was orally inoculated with P. gingivalis or vehicle only (sham). The P. gingivalis levels were determined by qPCR of the ISPg1 gene ( P. gingivalis ) at 2 weeks postinfection. There was no statistically significant difference in P. gingivalis levels among groups. P. gingivalis -specific antibody responses were determined at the termination of the experiment. The antibody titers in TLR9 −/− mice and WT mice infected with P. gingivalis were significantly higher than the titers in the uninfected mice ( P

    Techniques Used: Mouse Assay, Real-time Polymerase Chain Reaction, Infection

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

    Techniques Used: Enzyme-linked Immunosorbent Assay

    34) Product Images from "Serine dipeptide lipids of Porphyromonas gingivalis inhibit osteoblast differentiation: Relationship to Toll-like receptor 2"

    Article Title: Serine dipeptide lipids of Porphyromonas gingivalis inhibit osteoblast differentiation: Relationship to Toll-like receptor 2

    Journal: Bone

    doi: 10.1016/j.bone.2015.09.008

    Effects of P. gingivalis Lipid 654, Lipid 430 and total lipids on Col2.3GFP expression and mineral deposit formation in wild type and TLR2 KO calvarial cells. The results are depicted as the percent GFP or VK stained areas for the treated culture normalized
    Figure Legend Snippet: Effects of P. gingivalis Lipid 654, Lipid 430 and total lipids on Col2.3GFP expression and mineral deposit formation in wild type and TLR2 KO calvarial cells. The results are depicted as the percent GFP or VK stained areas for the treated culture normalized

    Techniques Used: Expressing, Staining

    Effect of P. gingivalis Lipid 654 and Lipid 430 on Col2.3GFP expression in vivo. Lipids were sonicated in PBS (30 sec, 3 watts). Col2.3GFP wild type or TLR2 KO:Col2.3GFP mice were lightly anesthetized with isofluorane and phosphate buffered saline (PBS
    Figure Legend Snippet: Effect of P. gingivalis Lipid 654 and Lipid 430 on Col2.3GFP expression in vivo. Lipids were sonicated in PBS (30 sec, 3 watts). Col2.3GFP wild type or TLR2 KO:Col2.3GFP mice were lightly anesthetized with isofluorane and phosphate buffered saline (PBS

    Techniques Used: Expressing, In Vivo, Sonication, Size-exclusion Chromatography, Mouse Assay

    Effects of P. gingivalis Lipid 654 and Lipid 430 on Col2.3GFP expression and mineral deposit formation in wild type and TLR2 KO calvarial cells. Calvarial cells were isolated as described in the Materials and Methods. Lipid preparations were sonicated
    Figure Legend Snippet: Effects of P. gingivalis Lipid 654 and Lipid 430 on Col2.3GFP expression and mineral deposit formation in wild type and TLR2 KO calvarial cells. Calvarial cells were isolated as described in the Materials and Methods. Lipid preparations were sonicated

    Techniques Used: Expressing, Isolation, Sonication

    35) Product Images from "Co-Localized or Randomly Distributed? Pair Cross Correlation of In Vivo Grown Subgingival Biofilm Bacteria Quantified by Digital Image Analysis"

    Article Title: Co-Localized or Randomly Distributed? Pair Cross Correlation of In Vivo Grown Subgingival Biofilm Bacteria Quantified by Digital Image Analysis

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0037583

    Pair cross correlation results. The mean PCC function g ( r) (continuous line) and the 95% confidence interval (dotted lines) are plotted against distances r spaced at intervals of ∼0.5 µm. The dashed horizontal reference line on the level of g ( r ) = 1 corresponds to the value of randomness and provides an internal ‘null hypothesis’ for testing attraction or repulsion between cellular units. (A) Representative, individual-related PCC of T. forsythia and F. nucleatum/periodonticum calculated for 25 images obtained from patient 01. A pronounced peak of 2.5 PCC values at 1.5 µm indicated co-localization of T. forsythia and F. nucleatum/periodonticum cells within short distances from 0–6 µm. (B) Outlier evaluation. PCC of T. forsythia and F. nucleatum/periodonticum calculated for 25 images obtained from patient 10. An initially prominent peak was in contrast to Figure 3A embedded in a wide CI, which lower boundary (−95%) dropped below the reference line, indicating a high variance in PCC values within the first 3 µm. (C) Representative, individual-related PCC of P. gingivalis and P. intermedia calculated for 32 images obtained from patient 05. P. gingivalis and P. intermedia cells are randomly distributed within the entire distance range. (D) Outlier evaluation. PCC of P. gingivalis and P. intermedia was calculated for 30 images obtained from patient 04. Similar to the outlier results of T. forsythia and F. nucleatum/periodonticum ( Figure 3 B), the high variance of PCC values at distances
    Figure Legend Snippet: Pair cross correlation results. The mean PCC function g ( r) (continuous line) and the 95% confidence interval (dotted lines) are plotted against distances r spaced at intervals of ∼0.5 µm. The dashed horizontal reference line on the level of g ( r ) = 1 corresponds to the value of randomness and provides an internal ‘null hypothesis’ for testing attraction or repulsion between cellular units. (A) Representative, individual-related PCC of T. forsythia and F. nucleatum/periodonticum calculated for 25 images obtained from patient 01. A pronounced peak of 2.5 PCC values at 1.5 µm indicated co-localization of T. forsythia and F. nucleatum/periodonticum cells within short distances from 0–6 µm. (B) Outlier evaluation. PCC of T. forsythia and F. nucleatum/periodonticum calculated for 25 images obtained from patient 10. An initially prominent peak was in contrast to Figure 3A embedded in a wide CI, which lower boundary (−95%) dropped below the reference line, indicating a high variance in PCC values within the first 3 µm. (C) Representative, individual-related PCC of P. gingivalis and P. intermedia calculated for 32 images obtained from patient 05. P. gingivalis and P. intermedia cells are randomly distributed within the entire distance range. (D) Outlier evaluation. PCC of P. gingivalis and P. intermedia was calculated for 30 images obtained from patient 04. Similar to the outlier results of T. forsythia and F. nucleatum/periodonticum ( Figure 3 B), the high variance of PCC values at distances

    Techniques Used: Periodic Counter-current Chromatography

    PCC curves of consolidated patient groups for T. forsythia versus F. nucleatum/periodonticum ( n = 8) and P. gingivalis versus P. intermedia ( n = 6). To compare the results of co-localization and randomness the patient group of each bacterial pair was merged by calculating the arithmetic mean curve with respective 95% CI by applying the equation 95% CI = m±1.96* SEM . The mean PCC curve for T. forsythia versus F. nucleatum/periodonticum (gray line, unfilled circles) and P. gingivalis /versus P. intermedia (black line, diamonds) were plotted with their respective 95% CI (dotted lines) against distances r in a range from 0–25 µm. The two curves were clearly distinguished from each other, by peak-levels and by convergence with the reference line. The fact that the lower CI of the co-localized bacteria was evidently separated from the upper CI of the randomly distributed pair of species within a wide range of 0–19 µm indicated a significant difference of both curves.
    Figure Legend Snippet: PCC curves of consolidated patient groups for T. forsythia versus F. nucleatum/periodonticum ( n = 8) and P. gingivalis versus P. intermedia ( n = 6). To compare the results of co-localization and randomness the patient group of each bacterial pair was merged by calculating the arithmetic mean curve with respective 95% CI by applying the equation 95% CI = m±1.96* SEM . The mean PCC curve for T. forsythia versus F. nucleatum/periodonticum (gray line, unfilled circles) and P. gingivalis /versus P. intermedia (black line, diamonds) were plotted with their respective 95% CI (dotted lines) against distances r in a range from 0–25 µm. The two curves were clearly distinguished from each other, by peak-levels and by convergence with the reference line. The fact that the lower CI of the co-localized bacteria was evidently separated from the upper CI of the randomly distributed pair of species within a wide range of 0–19 µm indicated a significant difference of both curves.

    Techniques Used: Periodic Counter-current Chromatography

    Visualization of the spatial arrangement of P. gingivalis and P. intermedia . FISH analysis performed on a carrier of GAP patient 05 reveals P. gingivalis in combination with P. intermedia respectively detected by the probes POGI-Cy5 (green) and PRIN-Cy3 (red). The domain-specific probe EUB338-FITC (blue) displays the entire biofilm expanded between the gingival surface at the bottom of the image and the carrier surface at the top. (A) An overlay of Cy3, Cy5 and FITC channels shows discrete microcolonies of P. gingivalis and P. intermedia apparently equispaced except the part marked by arrowheads. (B) In a punctual area species-specific channels Cy3 and Cy5 reveal both bacteria closely intermingled. (C–E) shows the respective binary masks of the micrograph (A) prepared for statistical quantification of the spatial relationship. (C) The EUB338-FITC-channel served as reference mask to limit the calculation to the area of the biomass. (D–E) Between the segmented masks of species-specific channels PRIN-Cy3 (red) and (E) POGI-Cy5 (green) daime calculated the pair cross correlation function g(r) .
    Figure Legend Snippet: Visualization of the spatial arrangement of P. gingivalis and P. intermedia . FISH analysis performed on a carrier of GAP patient 05 reveals P. gingivalis in combination with P. intermedia respectively detected by the probes POGI-Cy5 (green) and PRIN-Cy3 (red). The domain-specific probe EUB338-FITC (blue) displays the entire biofilm expanded between the gingival surface at the bottom of the image and the carrier surface at the top. (A) An overlay of Cy3, Cy5 and FITC channels shows discrete microcolonies of P. gingivalis and P. intermedia apparently equispaced except the part marked by arrowheads. (B) In a punctual area species-specific channels Cy3 and Cy5 reveal both bacteria closely intermingled. (C–E) shows the respective binary masks of the micrograph (A) prepared for statistical quantification of the spatial relationship. (C) The EUB338-FITC-channel served as reference mask to limit the calculation to the area of the biomass. (D–E) Between the segmented masks of species-specific channels PRIN-Cy3 (red) and (E) POGI-Cy5 (green) daime calculated the pair cross correlation function g(r) .

    Techniques Used: Fluorescence In Situ Hybridization

    36) Product Images from "Periodontal reconstruction by heparan sulfate mimetic-based matrix therapy in Porphyromonas gingivalis-infected mice"

    Article Title: Periodontal reconstruction by heparan sulfate mimetic-based matrix therapy in Porphyromonas gingivalis-infected mice

    Journal: Heliyon

    doi: 10.1016/j.heliyon.2018.e00719

    Timeline of the experimental protocol. After oral flora depression (black arrow) the mice were orally infected daily for 5 days with A. viscosus , strain CIP 103147 (blue arrows). Two weeks later (start of the experimental period), P. gingivalis was orally inoculated for five days during the first week (red arrows). For the following 11 weeks, P. gingivalis (strain W83) was inoculated 3 times per week (disease induction phase). At the end of the 12 th week, control (n = 15) and P gingivalis -inoculated mice (n = 15) were killed to evaluate the macroscopic alveolar bone status. The remaining periodontitis-affected mice were separated into 2 groups (treatment phase). One group (n = 15) was inoculated 3 times per week and received weakly saline intramuscular injections for 8 weeks (sham-treated periodontitis group). The other group (n = 30) was inoculated 3 times per week and received weekly intramuscular injections of RGTA in saline (1.5 mg/kg bw) for 8 weeks (RGTA-treated group (green arrows). At the end of the treatment period, the mice from the control (n = 15), sham-treated (n = 15) and RGTA-treated (n = 30) groups were killed.
    Figure Legend Snippet: Timeline of the experimental protocol. After oral flora depression (black arrow) the mice were orally infected daily for 5 days with A. viscosus , strain CIP 103147 (blue arrows). Two weeks later (start of the experimental period), P. gingivalis was orally inoculated for five days during the first week (red arrows). For the following 11 weeks, P. gingivalis (strain W83) was inoculated 3 times per week (disease induction phase). At the end of the 12 th week, control (n = 15) and P gingivalis -inoculated mice (n = 15) were killed to evaluate the macroscopic alveolar bone status. The remaining periodontitis-affected mice were separated into 2 groups (treatment phase). One group (n = 15) was inoculated 3 times per week and received weakly saline intramuscular injections for 8 weeks (sham-treated periodontitis group). The other group (n = 30) was inoculated 3 times per week and received weekly intramuscular injections of RGTA in saline (1.5 mg/kg bw) for 8 weeks (RGTA-treated group (green arrows). At the end of the treatment period, the mice from the control (n = 15), sham-treated (n = 15) and RGTA-treated (n = 30) groups were killed.

    Techniques Used: Mouse Assay, Infection

    37) Product Images from "Porphyromonas gingivalis in Alzheimer’s disease brains: Evidence for disease causation and treatment with small-molecule inhibitors"

    Article Title: Porphyromonas gingivalis in Alzheimer’s disease brains: Evidence for disease causation and treatment with small-molecule inhibitors

    Journal: Science Advances

    doi: 10.1126/sciadv.aau3333

    COR388 target engagement and dose-dependent effects on brain P. gingivalis , Aβ 1–42 , and TNFα in mice. ( A ) COR553 fluorescent activity probe for Kgp. ( B ) COR553 labeling of Kgp in P. gingivalis W83 strain and no labeling in mutant deficient in Kgp (ΔKgp). ( C ) W83 lysates labeled with COR553. Left lane, before immunodepletion; middle lane, after immunodepletion with anti-Kgp–conjugated beads; right lane, after elution from anti-Kgp–conjugated beads. ( D ) W83 strain titrated and labeled with COR553 to determine the limit of bacterial detection. See Results for details. ( E ) Oral plaque samples from human subjects (CB1-5) with periodontal disease were incubated ex vivo with COR553 probe with or without preincubation with COR388. COR553 probe and CAB102 detected Kgp strongly in three subjects (CB1, CB4, and CB5) and weakly in one subject (CB3). COR388 preincubation blocked COR553 probe binding to Kgp. ( F ) qPCR analysis of plaque samples using hmuY gene–specific primers identified P. gingivalis DNA in samples. ( G ) qPCR analysis of saliva samples. The bar graphs in (F) and (G) show the means and SEMs of three replicates. ( H ) COR388 treatment of W83 culture in defined growth medium reduced growth similarly to a Kgp-deficient strain (ΔKgp) over 43 hours. ( I ) Resistance developed rapidly to moxifloxacin but not COR388 with repeat passaging of bacterial culture. ( J to L ) Efficacy of COR388 at three oral doses of 3, 10, and 30 mg/kg twice daily in treating an established P. gingivalis brain infection in mice. Reduction of brain tissue levels of P. gingivalis (J), Aβ 1–42 (K), and TNFα (L). The bar graphs show the means with SEM error bars. *** P
    Figure Legend Snippet: COR388 target engagement and dose-dependent effects on brain P. gingivalis , Aβ 1–42 , and TNFα in mice. ( A ) COR553 fluorescent activity probe for Kgp. ( B ) COR553 labeling of Kgp in P. gingivalis W83 strain and no labeling in mutant deficient in Kgp (ΔKgp). ( C ) W83 lysates labeled with COR553. Left lane, before immunodepletion; middle lane, after immunodepletion with anti-Kgp–conjugated beads; right lane, after elution from anti-Kgp–conjugated beads. ( D ) W83 strain titrated and labeled with COR553 to determine the limit of bacterial detection. See Results for details. ( E ) Oral plaque samples from human subjects (CB1-5) with periodontal disease were incubated ex vivo with COR553 probe with or without preincubation with COR388. COR553 probe and CAB102 detected Kgp strongly in three subjects (CB1, CB4, and CB5) and weakly in one subject (CB3). COR388 preincubation blocked COR553 probe binding to Kgp. ( F ) qPCR analysis of plaque samples using hmuY gene–specific primers identified P. gingivalis DNA in samples. ( G ) qPCR analysis of saliva samples. The bar graphs in (F) and (G) show the means and SEMs of three replicates. ( H ) COR388 treatment of W83 culture in defined growth medium reduced growth similarly to a Kgp-deficient strain (ΔKgp) over 43 hours. ( I ) Resistance developed rapidly to moxifloxacin but not COR388 with repeat passaging of bacterial culture. ( J to L ) Efficacy of COR388 at three oral doses of 3, 10, and 30 mg/kg twice daily in treating an established P. gingivalis brain infection in mice. Reduction of brain tissue levels of P. gingivalis (J), Aβ 1–42 (K), and TNFα (L). The bar graphs show the means with SEM error bars. *** P

    Techniques Used: Mouse Assay, Activity Assay, Labeling, Mutagenesis, Incubation, Ex Vivo, Binding Assay, Real-time Polymerase Chain Reaction, Passaging, Infection

    Small-molecule gingipain inhibitors protect neuronal cells against P. gingivalis – and gingipain-induced toxicity in vitro and in vivo. ( A ) Differentiated SH-SY5Y neuroblastoma cells demonstrate cell aggregation after exposure to RgpB (10 μg/ml), Kgp (10 μg/ml), or both for 24 hours. The nonselective cysteine protease inhibitor iodoacetamide (IAM) blocks the gingipain-induced cell aggregation. ( B ) AlamarBlue viability assay shows that P. gingivalis ( P.g. ) is toxic to SH-SY5Y cells (MOI of 400) and that the small-molecule Kgp inhibitor COR271 and the RgpB inhibitor COR286 provide dose-dependent protection. The broad-spectrum antibiotics moxifloxacin and doxycycline and the γ-secretase inhibitor semagacestat did not inhibit the cytotoxic effect of P. gingivalis . ( C ) Fluoro-Jade C (FJC) staining (green) in pyramidal neurons of the CA1 region of the mouse hippocampus indicates neurodegeneration after stereotactic injection of gingipains. Counterstain with 4′,6-diamidino-2-phenylindole (DAPI) (blue). Scale bars, 50 μm. ( D ) The total number of FJC-positive cells was determined from serial section through the entire hippocampus. Results demonstrate a significant neuroprotective effect of gingipain inhibitors COR271 + COR286 after acute gingipain exposure in the hippocampus (* P
    Figure Legend Snippet: Small-molecule gingipain inhibitors protect neuronal cells against P. gingivalis – and gingipain-induced toxicity in vitro and in vivo. ( A ) Differentiated SH-SY5Y neuroblastoma cells demonstrate cell aggregation after exposure to RgpB (10 μg/ml), Kgp (10 μg/ml), or both for 24 hours. The nonselective cysteine protease inhibitor iodoacetamide (IAM) blocks the gingipain-induced cell aggregation. ( B ) AlamarBlue viability assay shows that P. gingivalis ( P.g. ) is toxic to SH-SY5Y cells (MOI of 400) and that the small-molecule Kgp inhibitor COR271 and the RgpB inhibitor COR286 provide dose-dependent protection. The broad-spectrum antibiotics moxifloxacin and doxycycline and the γ-secretase inhibitor semagacestat did not inhibit the cytotoxic effect of P. gingivalis . ( C ) Fluoro-Jade C (FJC) staining (green) in pyramidal neurons of the CA1 region of the mouse hippocampus indicates neurodegeneration after stereotactic injection of gingipains. Counterstain with 4′,6-diamidino-2-phenylindole (DAPI) (blue). Scale bars, 50 μm. ( D ) The total number of FJC-positive cells was determined from serial section through the entire hippocampus. Results demonstrate a significant neuroprotective effect of gingipain inhibitors COR271 + COR286 after acute gingipain exposure in the hippocampus (* P

    Techniques Used: In Vitro, In Vivo, Protease Inhibitor, Viability Assay, Staining, Injection

    RgpB colocalizes with neurons and pathology in AD hippocampus. ( A ) IHC using RgpB-specific monoclonal antibody 18E6 (representative images from a 63-year-old AD patient). The hippocampus shows abundant intracellular RgpB in the hilus (1), CA3 pyramidal layer (2), granular cell layer (3), and molecular layer (4). High-magnification images from the indicated areas (1 to 4) exhibit a granular staining pattern consistent with P. gingivalis intracellular infection. Scale bars, 200 μm (overview), 50 μm (1), and 10 μm (2 to 4). ( B ) AD hippocampus stained with 18E6 (AD) compared to gingival tissue (gingiva) from a patient with periodontal disease as well as a non-AD control and mouse IgG1 control (IgG1) in an adjacent hippocampal section. Scale bars, 50 μm. ( C ) Immunofluorescent colabeling with CAB101 reveals granular intraneuronal staining for RgpB (arrows) in MAP2-positive neurons in both the granular cell layer (GCL) and the pyramidal cell layer (CA1). Scale bars, 10 μm. ( D ) Dense extracellular RgpB-positive aggregates (arrowheads) were closely associated with astrocytes [glial fibrillary acidic protein (GFAP)]. There was no observed association of RgpB with microglia (IBA1). Scale bars, 10 μm. ( E ) RgpB was associated with paired helical filament Tau (PHF-Tau; arrows). RgpB-positive neurons negative for PHF-Tau (arrowheads) were also seen. Intracellular Aβ was often colocalized with RgpB (arrows). In some Aβ-positive cells, RgpB could not be detected (arrowheads). Scale bars, 10 μm.
    Figure Legend Snippet: RgpB colocalizes with neurons and pathology in AD hippocampus. ( A ) IHC using RgpB-specific monoclonal antibody 18E6 (representative images from a 63-year-old AD patient). The hippocampus shows abundant intracellular RgpB in the hilus (1), CA3 pyramidal layer (2), granular cell layer (3), and molecular layer (4). High-magnification images from the indicated areas (1 to 4) exhibit a granular staining pattern consistent with P. gingivalis intracellular infection. Scale bars, 200 μm (overview), 50 μm (1), and 10 μm (2 to 4). ( B ) AD hippocampus stained with 18E6 (AD) compared to gingival tissue (gingiva) from a patient with periodontal disease as well as a non-AD control and mouse IgG1 control (IgG1) in an adjacent hippocampal section. Scale bars, 50 μm. ( C ) Immunofluorescent colabeling with CAB101 reveals granular intraneuronal staining for RgpB (arrows) in MAP2-positive neurons in both the granular cell layer (GCL) and the pyramidal cell layer (CA1). Scale bars, 10 μm. ( D ) Dense extracellular RgpB-positive aggregates (arrowheads) were closely associated with astrocytes [glial fibrillary acidic protein (GFAP)]. There was no observed association of RgpB with microglia (IBA1). Scale bars, 10 μm. ( E ) RgpB was associated with paired helical filament Tau (PHF-Tau; arrows). RgpB-positive neurons negative for PHF-Tau (arrowheads) were also seen. Intracellular Aβ was often colocalized with RgpB (arrows). In some Aβ-positive cells, RgpB could not be detected (arrowheads). Scale bars, 10 μm.

    Techniques Used: Immunohistochemistry, Staining, Infection

    Identification of P. gingivalis –specific protein and DNA in cortex from control and AD patients. ( A ) WB with four different strains of P. gingivalis and CAB102 detection of typical molecular weight bands for Kgp in bacterial lysates. ( B ) IP using brain lysates from nondemented controls (C1 to C6; ages 75, 54, 63, 45, 37, and 102 years, respectively) and AD patients (AD1 to AD3; ages 83, 90, and 80 years, respectively) using CAB102 with subsequent WB reveals the ~50-kDa Kgp catalytic subunit (Kgp cat ), along with higher– and lower–molecular weight Kgp species seen in (A). ( C ) qPCR from DNA isolated from the same brain lysates as the protein samples analyzed in (B) shows a positive signal in nondemented control (C1 to C5) and AD (AD1 to AD3) samples. Sample C6 from the 102-year-old nondemented control patient had no detectable qPCR signal in (C) and very faint bands indicating near absence of Kgp (B) (mean with SEM error bars of repeat qPCR runs).
    Figure Legend Snippet: Identification of P. gingivalis –specific protein and DNA in cortex from control and AD patients. ( A ) WB with four different strains of P. gingivalis and CAB102 detection of typical molecular weight bands for Kgp in bacterial lysates. ( B ) IP using brain lysates from nondemented controls (C1 to C6; ages 75, 54, 63, 45, 37, and 102 years, respectively) and AD patients (AD1 to AD3; ages 83, 90, and 80 years, respectively) using CAB102 with subsequent WB reveals the ~50-kDa Kgp catalytic subunit (Kgp cat ), along with higher– and lower–molecular weight Kgp species seen in (A). ( C ) qPCR from DNA isolated from the same brain lysates as the protein samples analyzed in (B) shows a positive signal in nondemented control (C1 to C5) and AD (AD1 to AD3) samples. Sample C6 from the 102-year-old nondemented control patient had no detectable qPCR signal in (C) and very faint bands indicating near absence of Kgp (B) (mean with SEM error bars of repeat qPCR runs).

    Techniques Used: Western Blot, Molecular Weight, Real-time Polymerase Chain Reaction, Isolation

    Detection of P. gingivalis in CSF and oral biofluids from clinical AD subjects. ( A ) Detection and quantitation of P. gingivalis DNA by qPCR in CSF from subjects with probable AD. ( B ) Detection and quantitation of P. gingivalis DNA by qPCR from matching saliva samples. ( C ) Top: PCR products detecting P. gingivalis from CSF in (A) from all subjects run on agarose gel including negative and positive controls containing a synthetic DNA template. Faint or undetectable PCR products from subjects AD1, AD3, and AD5 were below the limit of quantitation for copy number and not of sufficient quantity for sequence analysis. Bottom: qPCR products from CSF from the same subjects for H. pylori. ( D ) Data table includes age and Mini Mental Status Exam (MMSE) score on subjects and sequence identity of PCR products to P. gingivalis hmuY DNA sequence. Sequence data are included in fig. S4. NS, not sequenced.
    Figure Legend Snippet: Detection of P. gingivalis in CSF and oral biofluids from clinical AD subjects. ( A ) Detection and quantitation of P. gingivalis DNA by qPCR in CSF from subjects with probable AD. ( B ) Detection and quantitation of P. gingivalis DNA by qPCR from matching saliva samples. ( C ) Top: PCR products detecting P. gingivalis from CSF in (A) from all subjects run on agarose gel including negative and positive controls containing a synthetic DNA template. Faint or undetectable PCR products from subjects AD1, AD3, and AD5 were below the limit of quantitation for copy number and not of sufficient quantity for sequence analysis. Bottom: qPCR products from CSF from the same subjects for H. pylori. ( D ) Data table includes age and Mini Mental Status Exam (MMSE) score on subjects and sequence identity of PCR products to P. gingivalis hmuY DNA sequence. Sequence data are included in fig. S4. NS, not sequenced.

    Techniques Used: Quantitation Assay, Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Sequencing

    P. gingivalis invasion of the brain induces an Aβ 1–42 response that is blocked by gingipain inhibition in mice. ( A ) P. gingivalis PCR product in mouse brains after oral infection with P. gingivalis W83, with or without treatment with the Kgp inhibitor COR119, or infection with gingipain knockout strain ΔRgpB or ΔKgp. Lanes 1 to 8 represent individual experimental animals. In the first lane ( P.g. ), P. gingivalis W83 was used as a positive control. ( B ) P. gingivalis W83–infected mice, but not COR119-treated mice or mice infected with gingipain knockouts, had significantly higher Aβ 1–42 levels compared to mock-infected mice (*** P
    Figure Legend Snippet: P. gingivalis invasion of the brain induces an Aβ 1–42 response that is blocked by gingipain inhibition in mice. ( A ) P. gingivalis PCR product in mouse brains after oral infection with P. gingivalis W83, with or without treatment with the Kgp inhibitor COR119, or infection with gingipain knockout strain ΔRgpB or ΔKgp. Lanes 1 to 8 represent individual experimental animals. In the first lane ( P.g. ), P. gingivalis W83 was used as a positive control. ( B ) P. gingivalis W83–infected mice, but not COR119-treated mice or mice infected with gingipain knockouts, had significantly higher Aβ 1–42 levels compared to mock-infected mice (*** P

    Techniques Used: Inhibition, Mouse Assay, Polymerase Chain Reaction, Infection, Knock-Out, Positive Control

    P. gingivalis and gingipains fragment tau. ( A ) WB analysis of total soluble tau in SH-SY5Y cells infected with increasing concentrations of wild-type (WT) P. gingivalis strain W83 ( P.g. ) and P. gingivalis gingipain-deficient mutants either lacking Kgp activity (KgpΔIg-B) or lacking both Kgp and Rgp activity (ΔK/ΔRAB-A) . Uninfected SH-SY5Y cells (No P.g. ) were used as a negative control. Glyceraldehyde-phosphate dehydrogenase (GAPDH) was used as a loading control. Total tau was monitored with the monoclonal antibody Tau-5 at 1, 4, and 8 hours after infection. ( B ) Densitometry analysis of the total tau WB images. ( C ) WB analysis of rtau-441 incubated with purified Kgp and RgpB catalytic domains combined (Gp) at various concentrations for 1 hour at 37°C. The blot was probed with tau monoclonal antibody T46. ( D ) Gingipain cleavage sites in rtau-441 deduced from peptide fragments identified by MS for rtau-441 incubated with 1 or 10 nM gingipains. (a) T46 antibody epitope (red). (b) Tau-5 antibody epitope (red). (c) N-terminal tau fragment. (d) C-terminal tau fragment. (e) Kgp-generated tau fragments containing the VQIVYK sequence. (f) Kgp-generated fragments containing the VQIINK sequence. (g) An RgpB-generated tau fragment. *Cleavage sites identified at 1 nM gingipains.
    Figure Legend Snippet: P. gingivalis and gingipains fragment tau. ( A ) WB analysis of total soluble tau in SH-SY5Y cells infected with increasing concentrations of wild-type (WT) P. gingivalis strain W83 ( P.g. ) and P. gingivalis gingipain-deficient mutants either lacking Kgp activity (KgpΔIg-B) or lacking both Kgp and Rgp activity (ΔK/ΔRAB-A) . Uninfected SH-SY5Y cells (No P.g. ) were used as a negative control. Glyceraldehyde-phosphate dehydrogenase (GAPDH) was used as a loading control. Total tau was monitored with the monoclonal antibody Tau-5 at 1, 4, and 8 hours after infection. ( B ) Densitometry analysis of the total tau WB images. ( C ) WB analysis of rtau-441 incubated with purified Kgp and RgpB catalytic domains combined (Gp) at various concentrations for 1 hour at 37°C. The blot was probed with tau monoclonal antibody T46. ( D ) Gingipain cleavage sites in rtau-441 deduced from peptide fragments identified by MS for rtau-441 incubated with 1 or 10 nM gingipains. (a) T46 antibody epitope (red). (b) Tau-5 antibody epitope (red). (c) N-terminal tau fragment. (d) C-terminal tau fragment. (e) Kgp-generated tau fragments containing the VQIVYK sequence. (f) Kgp-generated fragments containing the VQIINK sequence. (g) An RgpB-generated tau fragment. *Cleavage sites identified at 1 nM gingipains.

    Techniques Used: Western Blot, Infection, Activity Assay, Negative Control, Incubation, Purification, Mass Spectrometry, Generated, Sequencing

    38) Product Images from "Purification and Characterization of Arginine Carboxypeptidase Produced by Porphyromonas gingivalis"

    Article Title: Purification and Characterization of Arginine Carboxypeptidase Produced by Porphyromonas gingivalis

    Journal: Infection and Immunity

    doi: 10.1128/IAI.70.4.1807-1815.2002

    Suspected RCP sequence from the unfinished P. gingivalis W83 genome. The ORF of the DNA sequence and the deduced amino acid sequence are shown. The underlined amino acid sequence was determined by sequencing the N terminus of the isolated protein. The consensus signature sequence of the zinc-binding region conserved in zinc carboxypeptidases is doubly underlined. Potential N-linked glycosylation sites are boxed.
    Figure Legend Snippet: Suspected RCP sequence from the unfinished P. gingivalis W83 genome. The ORF of the DNA sequence and the deduced amino acid sequence are shown. The underlined amino acid sequence was determined by sequencing the N terminus of the isolated protein. The consensus signature sequence of the zinc-binding region conserved in zinc carboxypeptidases is doubly underlined. Potential N-linked glycosylation sites are boxed.

    Techniques Used: Sequencing, Isolation, Binding Assay

    Western blotting analysis of cell fractions of P. gingivalis . Western blotting was performed with an antibody against RCP (1:1,000 dilution). Following incubation with the antibody, the nitrocellulose membrane was reacted with a goat anti-rabbit antibody conjugated to horseradish peroxidase (1:3,000 dilution). Lanes: 1, periplasm (30 μg); 2, cytoplasm (30 μg); 3, cytoplasmic membrane-enriched cell envelope (30 μg); 4, outer membrane-enriched cell envelope (30 μg); 5, purified RCP (14 μg). The values on the left are molecular sizes in kilodaltons.
    Figure Legend Snippet: Western blotting analysis of cell fractions of P. gingivalis . Western blotting was performed with an antibody against RCP (1:1,000 dilution). Following incubation with the antibody, the nitrocellulose membrane was reacted with a goat anti-rabbit antibody conjugated to horseradish peroxidase (1:3,000 dilution). Lanes: 1, periplasm (30 μg); 2, cytoplasm (30 μg); 3, cytoplasmic membrane-enriched cell envelope (30 μg); 4, outer membrane-enriched cell envelope (30 μg); 5, purified RCP (14 μg). The values on the left are molecular sizes in kilodaltons.

    Techniques Used: Western Blot, Incubation, Purification

    39) Product Images from "Porphyromonas gulae Has Virulence and Immunological Characteristics Similar to Those of the Human Periodontal Pathogen Porphyromonas gingivalis"

    Article Title: Porphyromonas gulae Has Virulence and Immunological Characteristics Similar to Those of the Human Periodontal Pathogen Porphyromonas gingivalis

    Journal: Infection and Immunity

    doi: 10.1128/IAI.01500-15

    P. gulae is phagocytosed more readily than P. gingivalis. P. gingivalis (W50 and ATCC 33277) and P. gulae ATCC 51700 were labeled with pHrodo and examined for their ability to be phagocytosed by mouse macrophages (RAW 264.7) at increasing bacterium/cell
    Figure Legend Snippet: P. gulae is phagocytosed more readily than P. gingivalis. P. gingivalis (W50 and ATCC 33277) and P. gulae ATCC 51700 were labeled with pHrodo and examined for their ability to be phagocytosed by mouse macrophages (RAW 264.7) at increasing bacterium/cell

    Techniques Used: Labeling

    Macrophage cytokine response to P. gulae . RAW 264.7 macrophages were incubated with no bacteria (▩), P. gulae ATCC 51700 (□), P. gingivalis ATCC 33277 (), or P. gingivalis W50 (■) for 90 min before the cells were centrifuged and
    Figure Legend Snippet: Macrophage cytokine response to P. gulae . RAW 264.7 macrophages were incubated with no bacteria (▩), P. gulae ATCC 51700 (□), P. gingivalis ATCC 33277 (), or P. gingivalis W50 (■) for 90 min before the cells were centrifuged and

    Techniques Used: Incubation

    40) Product Images from "Porphyromonas gulae Has Virulence and Immunological Characteristics Similar to Those of the Human Periodontal Pathogen Porphyromonas gingivalis"

    Article Title: Porphyromonas gulae Has Virulence and Immunological Characteristics Similar to Those of the Human Periodontal Pathogen Porphyromonas gingivalis

    Journal: Infection and Immunity

    doi: 10.1128/IAI.01500-15

    P. gulae is phagocytosed more readily than P. gingivalis. P. gingivalis (W50 and ATCC 33277) and P. gulae ATCC 51700 were labeled with pHrodo and examined for their ability to be phagocytosed by mouse macrophages (RAW 264.7) at increasing bacterium/cell
    Figure Legend Snippet: P. gulae is phagocytosed more readily than P. gingivalis. P. gingivalis (W50 and ATCC 33277) and P. gulae ATCC 51700 were labeled with pHrodo and examined for their ability to be phagocytosed by mouse macrophages (RAW 264.7) at increasing bacterium/cell

    Techniques Used: Labeling

    Macrophage cytokine response to P. gulae . RAW 264.7 macrophages were incubated with no bacteria (▩), P. gulae ATCC 51700 (□), P. gingivalis ATCC 33277 (), or P. gingivalis W50 (■) for 90 min before the cells were centrifuged and
    Figure Legend Snippet: Macrophage cytokine response to P. gulae . RAW 264.7 macrophages were incubated with no bacteria (▩), P. gulae ATCC 51700 (□), P. gingivalis ATCC 33277 (), or P. gingivalis W50 (■) for 90 min before the cells were centrifuged and

    Techniques Used: Incubation

    Related Articles

    Electroporation:

    Article Title: Three CoA Transferases Involved in the Production of Short Chain Fatty Acids in Porphyromonas gingivalis
    Article Snippet: .. Ligated fragments were introduced into P. gingivalis ATCC 33277 by electroporation. .. The constructed mutants were designated PGAGU101 (PGN_1171::erm ), PGAGU104 (PGN_0725::erm ), PGAGU108 (PGN_1888::erm ), and PGAGU109 (PGN_1341::erm ).

    Labeling:

    Article Title: The Porphyromonas gingivalis Ferric Uptake Regulator Orthologue Binds Hemin and Regulates Hemin-Responsive Biofilm Development
    Article Snippet: .. A total of 6 paired microarray hybridizations were performed representing 6 biological replicates, where a balanced dye design was used, with the overall analyses including three microarrays where P. gingivalis ATCC 33277 samples were labeled with Cy3 and the paired ECR455 samples were labeled with Cy5 and three other microarrays where samples were labeled with the opposite combination of fluorophores. ..

    Enzyme-linked Immunosorbent Assay:

    Article Title: HUMORAL IMMUNE RESPONSE TO ANTIGENS OF Porphyromonas gingivalis ATCC 33277 IN CHRONIC PERIODONTITIS
    Article Snippet: .. Serum IgG levels against the cell sonicate antigen from P. gingivalis ATCC 33277 of 28 patients with chronic periodontitis (CP), 10 patients with gingivitis (G) and 21 periodontally healthy individuals (H) were measured by ELISA and Western immunoblotting. .. In the CP group, sera reactivity by ELISA was significantly higher than in the G and H groups (Kruskal-Wallis p < 0.001; Dunnet t3 p= 0.001 and Dunnet t3 p= 0.0001).

    Microarray:

    Article Title: The Porphyromonas gingivalis Ferric Uptake Regulator Orthologue Binds Hemin and Regulates Hemin-Responsive Biofilm Development
    Article Snippet: .. A total of 6 paired microarray hybridizations were performed representing 6 biological replicates, where a balanced dye design was used, with the overall analyses including three microarrays where P. gingivalis ATCC 33277 samples were labeled with Cy3 and the paired ECR455 samples were labeled with Cy5 and three other microarrays where samples were labeled with the opposite combination of fluorophores. ..

    other:

    Article Title: Identification of Dipeptidyl-Peptidase (DPP)5 and DPP7 in Porphyromonas endodontalis, Distinct from Those in Porphyromonas gingivalis
    Article Snippet: Bacterial strains and culture conditions P. endodontalis ATCC 35406 and P. gingivalis ATCC 33277 and ATCC 49417 were obtained from American Type Culture Collection.

    Article Title: Inhibitory Effect of Enterococcus faecium WB2000 on Volatile Sulfur Compound Production by Porphyromonas gingivalis
    Article Snippet: In mixed cultures with the other three lactic acid bacteria (S. salivarius JCM 5707, L. salivarius CIP 103140, and L. reuteri JCM 1112), the number of viable P. gingivalis ATCC 33277 decreased to less than the detection limit at 12 h. In mixed culture with P. gingivalis ATCC 33277, E. faecium WB2000 grew more rapidly than it did in mixed cultures with the other three lactic acid bacteria, and its growth plateaued at 6 h ( ).

    Article Title: Identification of genes required for the survival of B. fragilis using massive parallel sequencing of a saturated transposon mutant library
    Article Snippet: In addition, all six genes involved in lipid-A biosynthesis are essential for B. fragilis 638R and P. gingivalis ATCC 33277 (lpxA , lpxC, lpxD, BF638R_0493, lpxB , and BF638R_3307), however only the latter three genes are essential in B. thetaiotaomicron VPI-5482 even though all of the six genes are present only in a single copy in the B. thetaiotaomicron VPI-5482 genome [ ].

    Sequencing:

    Article Title: Purification and characterization of a novel secondary fimbrial protein from Porphyromonas gulae
    Article Snippet: .. However, the deduced amino acid sequence encoding the 67-kDa fimbrial protein of P. gingivalis ATCC 33277 showed 34% similarity with the 53-kDa fimbrial protein of P. gulae ATCC 51700 ( ). .. The amino acid sequence of the N -terminal 15 residues of the 53-kDa fimbrial protein of P. gulae ATCC 51700 (AGDNDYNHVGEYGGI) showed 12 of 15 residues identical to the 53-kDa fimbrial protein of P. gingivalis strain 381 (AGDNDYNPIGEYGGV) and 4 of 15 residues identical to the 67-kDa fimbrial protein of P. gingivalis ATCC 33277 (AGDGQDQANPDYHYV) but only 1 of 15 residues identical to the 41-kDa fimbrial protein of P. gulae ATCC 51700 (AFGVADDEAKVAKLT).

    Western Blot:

    Article Title: HUMORAL IMMUNE RESPONSE TO ANTIGENS OF Porphyromonas gingivalis ATCC 33277 IN CHRONIC PERIODONTITIS
    Article Snippet: .. Serum IgG levels against the cell sonicate antigen from P. gingivalis ATCC 33277 of 28 patients with chronic periodontitis (CP), 10 patients with gingivitis (G) and 21 periodontally healthy individuals (H) were measured by ELISA and Western immunoblotting. .. In the CP group, sera reactivity by ELISA was significantly higher than in the G and H groups (Kruskal-Wallis p < 0.001; Dunnet t3 p= 0.001 and Dunnet t3 p= 0.0001).

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    ATCC p gingivalis genomes
    Clustering by spacer content in CRISPR type 36.2 of Porphyromonas <t>gingivalis</t> . In type 36.2, the presence of each unique spacer is shown using a heatmap. The dendrogram was constructed from Euclidian distances. In the heatmap, the boxes indicate unique spacers and are arrayed horizontally. In the heatmap, 2 colors were used according to the bit score; red: ≥50, yellow:
    P Gingivalis Genomes, supplied by ATCC, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC p gingivalis
    Attachment of T. forsythia 43037 (Tf) and BFM571 to KB cells in the absence and presence of P. <t>gingivalis</t> OMVs (Pgv). Attachment levels (means ± standard errors) were expressed as the percentage of attached bacteria relative to the total number
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    ATCC p gingivalis omvs
    Attachment of T. forsythia 43037 (Tf) and BFM571 to KB cells in the absence and presence of P. <t>gingivalis</t> <t>OMVs</t> (Pgv). Attachment levels (means ± standard errors) were expressed as the percentage of attached bacteria relative to the total number
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    Clustering by spacer content in CRISPR type 36.2 of Porphyromonas gingivalis . In type 36.2, the presence of each unique spacer is shown using a heatmap. The dendrogram was constructed from Euclidian distances. In the heatmap, the boxes indicate unique spacers and are arrayed horizontally. In the heatmap, 2 colors were used according to the bit score; red: ≥50, yellow:

    Journal: Genome Biology and Evolution

    Article Title: CRISPR Regulation of Intraspecies Diversification by Limiting IS Transposition and Intercellular Recombination

    doi: 10.1093/gbe/evt075

    Figure Lengend Snippet: Clustering by spacer content in CRISPR type 36.2 of Porphyromonas gingivalis . In type 36.2, the presence of each unique spacer is shown using a heatmap. The dendrogram was constructed from Euclidian distances. In the heatmap, the boxes indicate unique spacers and are arrayed horizontally. In the heatmap, 2 colors were used according to the bit score; red: ≥50, yellow:

    Article Snippet: Our preliminary investigation of CRISPRs in three P. gingivalis strains demonstrated that CRISPR spacers exhibiting high nucleotide similarity to regions of P. gingivalis genomes were present and the number of spacers was diverse among the three genomes (TDC60: 89; W83: 44; and ATCC 33277: 137).

    Techniques: CRISPR, Construct

    Characteristics of recombination breakpoints among three Porphyromonas gingivalis genomes. ( A ) Fragments are shown in the alignment of two genome sequences (TDC60, ATCC 33277). The positions of MGEs or rRNA operons in the breakpoint gaps are indicated by colored broken lines, connecting the gaps and the bars (indicating the positions of the features on the genome), which are arrayed along the outside of the plot area. The red boxes on the plot area are the regions shown in ( B ) in detail. ( B ) Breakpoint gaps of TDC60 are enlarged in light gray areas surrounded by broken lines. The regions of ATCC 33277, which correspond to the enlarged gap of TDC60, are enlarged similarly. The fragments in TDC60 and ATCC 33277 are colored by red and deep blue, respectively. The regions exhibiting high nucleotide similarity to each other are shown by a yellow belt between two fragments. The 3-kb regions of the breakpoints are indicated by dark gray rectangles on the upper or lower side of the fragments. (i) rRNA operons in the breakpoint gap. The black arrows indicate rRNA genes. The light blue-filled boxes with arrows inside indicate ISs. (ii) ISs in the breakpoint gap. ( C ) The number of each feature in the breakpoint gap is plotted. The regions without any characteristic features are included under “Others.” The mean and standard deviations are provided by the horizontal and vertical lines, respectively. Statistical significance is indicated by an asterisk ( P

    Journal: Genome Biology and Evolution

    Article Title: CRISPR Regulation of Intraspecies Diversification by Limiting IS Transposition and Intercellular Recombination

    doi: 10.1093/gbe/evt075

    Figure Lengend Snippet: Characteristics of recombination breakpoints among three Porphyromonas gingivalis genomes. ( A ) Fragments are shown in the alignment of two genome sequences (TDC60, ATCC 33277). The positions of MGEs or rRNA operons in the breakpoint gaps are indicated by colored broken lines, connecting the gaps and the bars (indicating the positions of the features on the genome), which are arrayed along the outside of the plot area. The red boxes on the plot area are the regions shown in ( B ) in detail. ( B ) Breakpoint gaps of TDC60 are enlarged in light gray areas surrounded by broken lines. The regions of ATCC 33277, which correspond to the enlarged gap of TDC60, are enlarged similarly. The fragments in TDC60 and ATCC 33277 are colored by red and deep blue, respectively. The regions exhibiting high nucleotide similarity to each other are shown by a yellow belt between two fragments. The 3-kb regions of the breakpoints are indicated by dark gray rectangles on the upper or lower side of the fragments. (i) rRNA operons in the breakpoint gap. The black arrows indicate rRNA genes. The light blue-filled boxes with arrows inside indicate ISs. (ii) ISs in the breakpoint gap. ( C ) The number of each feature in the breakpoint gap is plotted. The regions without any characteristic features are included under “Others.” The mean and standard deviations are provided by the horizontal and vertical lines, respectively. Statistical significance is indicated by an asterisk ( P

    Article Snippet: Our preliminary investigation of CRISPRs in three P. gingivalis strains demonstrated that CRISPR spacers exhibiting high nucleotide similarity to regions of P. gingivalis genomes were present and the number of spacers was diverse among the three genomes (TDC60: 89; W83: 44; and ATCC 33277: 137).

    Techniques:

    Split network of 60 Porphyromonas gingivalis isolates obtained from concatenated seven loci sequences. A split network tree based upon the MLST data is shown. Circles indicate external nodes (each isolate) and are colored according to geographic origin (black: Japan; outlined: overseas or unspecified). fimA types are shown by light gray shadows. The numbers outside the isolate’s name indicate the patient source. Eleven colors are used to emphasize the clusters.

    Journal: Genome Biology and Evolution

    Article Title: CRISPR Regulation of Intraspecies Diversification by Limiting IS Transposition and Intercellular Recombination

    doi: 10.1093/gbe/evt075

    Figure Lengend Snippet: Split network of 60 Porphyromonas gingivalis isolates obtained from concatenated seven loci sequences. A split network tree based upon the MLST data is shown. Circles indicate external nodes (each isolate) and are colored according to geographic origin (black: Japan; outlined: overseas or unspecified). fimA types are shown by light gray shadows. The numbers outside the isolate’s name indicate the patient source. Eleven colors are used to emphasize the clusters.

    Article Snippet: Our preliminary investigation of CRISPRs in three P. gingivalis strains demonstrated that CRISPR spacers exhibiting high nucleotide similarity to regions of P. gingivalis genomes were present and the number of spacers was diverse among the three genomes (TDC60: 89; W83: 44; and ATCC 33277: 137).

    Techniques:

    Regions exhibiting high nucleotide similarity to P. gingivalis CRISPR spacers. Two examples of the 19 spacers exhibiting high nucleotide similarity to the P. gingivalis genome are shown. The white and black arrows indicate CDSs and rRNA genes, respectively. The arrows within the light blue-filled boxes indicate ISs. The orange regions indicate the sequences exhibiting high nucleotide similarity to CRISPR spacers. (i) Region exhibiting high nucleotide similarity to spacer 37_259: the transposase gene in IS Pg2 , in the TDC60 genome. (ii) Region exhibiting high nucleotide similarity to spacer 37_90: close to the IS both 2-kb upstream and 2-kb downstream in the 3 genomes.

    Journal: Genome Biology and Evolution

    Article Title: CRISPR Regulation of Intraspecies Diversification by Limiting IS Transposition and Intercellular Recombination

    doi: 10.1093/gbe/evt075

    Figure Lengend Snippet: Regions exhibiting high nucleotide similarity to P. gingivalis CRISPR spacers. Two examples of the 19 spacers exhibiting high nucleotide similarity to the P. gingivalis genome are shown. The white and black arrows indicate CDSs and rRNA genes, respectively. The arrows within the light blue-filled boxes indicate ISs. The orange regions indicate the sequences exhibiting high nucleotide similarity to CRISPR spacers. (i) Region exhibiting high nucleotide similarity to spacer 37_259: the transposase gene in IS Pg2 , in the TDC60 genome. (ii) Region exhibiting high nucleotide similarity to spacer 37_90: close to the IS both 2-kb upstream and 2-kb downstream in the 3 genomes.

    Article Snippet: Our preliminary investigation of CRISPRs in three P. gingivalis strains demonstrated that CRISPR spacers exhibiting high nucleotide similarity to regions of P. gingivalis genomes were present and the number of spacers was diverse among the three genomes (TDC60: 89; W83: 44; and ATCC 33277: 137).

    Techniques: CRISPR

    Spacer contents of Porphyromonas gingivalis isolates from seven patients in four CRISPR loci. Spacer arrays of 26 isolates from 7 patients are shown at each CRISPR locus. Each box indicates one spacer. The spacers in the arrays exhibit high nucleotide similarity to each other among the isolates if they are aligned vertically and have the same color. Blank boxes indicate absent spacers in the particular isolates. In patient no. 2, two colors are used because the D5 isolate has a type 30 spacer array that is distinct from those of D8 and D9. The spacers in type 36.2, shared among seven isolates of three patients, are indicated by deep yellow boxes and emphasized by dark gray belts.

    Journal: Genome Biology and Evolution

    Article Title: CRISPR Regulation of Intraspecies Diversification by Limiting IS Transposition and Intercellular Recombination

    doi: 10.1093/gbe/evt075

    Figure Lengend Snippet: Spacer contents of Porphyromonas gingivalis isolates from seven patients in four CRISPR loci. Spacer arrays of 26 isolates from 7 patients are shown at each CRISPR locus. Each box indicates one spacer. The spacers in the arrays exhibit high nucleotide similarity to each other among the isolates if they are aligned vertically and have the same color. Blank boxes indicate absent spacers in the particular isolates. In patient no. 2, two colors are used because the D5 isolate has a type 30 spacer array that is distinct from those of D8 and D9. The spacers in type 36.2, shared among seven isolates of three patients, are indicated by deep yellow boxes and emphasized by dark gray belts.

    Article Snippet: Our preliminary investigation of CRISPRs in three P. gingivalis strains demonstrated that CRISPR spacers exhibiting high nucleotide similarity to regions of P. gingivalis genomes were present and the number of spacers was diverse among the three genomes (TDC60: 89; W83: 44; and ATCC 33277: 137).

    Techniques: CRISPR

    Attachment of T. forsythia 43037 (Tf) and BFM571 to KB cells in the absence and presence of P. gingivalis OMVs (Pgv). Attachment levels (means ± standard errors) were expressed as the percentage of attached bacteria relative to the total number

    Journal:

    Article Title: Porphyromonas gingivalis Vesicles Enhance Attachment, and the Leucine-Rich Repeat BspA Protein Is Required for Invasion of Epithelial Cells by "Tannerella forsythia"

    doi: 10.1128/IAI.00062-06

    Figure Lengend Snippet: Attachment of T. forsythia 43037 (Tf) and BFM571 to KB cells in the absence and presence of P. gingivalis OMVs (Pgv). Attachment levels (means ± standard errors) were expressed as the percentage of attached bacteria relative to the total number

    Article Snippet: There were no significant differences between P. gingivalis - or P. gingivalis OMV-induced aggregations of T. forsythia ATCC 43037 and those of the BspA-defective mutant BFM571.

    Techniques:

    Coaggregation of T. forsythia in the presence of P. gingivalis and outer membrane vesicles purified from P. gingivalis . Data are representative of three independent experiments. Data points represent means ± standard errors of triplicate readings.

    Journal:

    Article Title: Porphyromonas gingivalis Vesicles Enhance Attachment, and the Leucine-Rich Repeat BspA Protein Is Required for Invasion of Epithelial Cells by "Tannerella forsythia"

    doi: 10.1128/IAI.00062-06

    Figure Lengend Snippet: Coaggregation of T. forsythia in the presence of P. gingivalis and outer membrane vesicles purified from P. gingivalis . Data are representative of three independent experiments. Data points represent means ± standard errors of triplicate readings.

    Article Snippet: There were no significant differences between P. gingivalis - or P. gingivalis OMV-induced aggregations of T. forsythia ATCC 43037 and those of the BspA-defective mutant BFM571.

    Techniques: Purification

    Invasion by T. forsythia (Tf) of KB cells and dependence of invasion on the presence of P. gingivalis vesicles (Pgv). The invasion level was expressed as the percentage of bacteria retrieved following antibiotic killing of external bacteria and KB cell

    Journal:

    Article Title: Porphyromonas gingivalis Vesicles Enhance Attachment, and the Leucine-Rich Repeat BspA Protein Is Required for Invasion of Epithelial Cells by "Tannerella forsythia"

    doi: 10.1128/IAI.00062-06

    Figure Lengend Snippet: Invasion by T. forsythia (Tf) of KB cells and dependence of invasion on the presence of P. gingivalis vesicles (Pgv). The invasion level was expressed as the percentage of bacteria retrieved following antibiotic killing of external bacteria and KB cell

    Article Snippet: There were no significant differences between P. gingivalis - or P. gingivalis OMV-induced aggregations of T. forsythia ATCC 43037 and those of the BspA-defective mutant BFM571.

    Techniques:

    Effect of serum on viability of A. oris (Ao), V. parvula (Vp), F. nucleatum ATCC 10953 (Fn) and P. gingivalis 381 (Pg). Planktonic growth of microorganisms was evaluated in their appropriate nutrient-rich medium supplemented with 10% native human serum

    Journal: Anaerobe

    Article Title: Strain-Specific Colonization Patterns and Serum Modulation of Multi-Species Oral Biofilm Development

    doi: 10.1016/j.anaerobe.2012.06.003

    Figure Lengend Snippet: Effect of serum on viability of A. oris (Ao), V. parvula (Vp), F. nucleatum ATCC 10953 (Fn) and P. gingivalis 381 (Pg). Planktonic growth of microorganisms was evaluated in their appropriate nutrient-rich medium supplemented with 10% native human serum

    Article Snippet: For these experiments, we inoculated flow cells simultaneously with A. oris , V. parvula , F. nucleatum ATCC 10953 and P. gingivalis 381. shows the effect of serum on multi-species biofilms and also on mono-species biofilms of F. nucleatum ATCC 10953 and P. gingivalis 381.

    Techniques:

    Attachment of T. forsythia 43037 (Tf) and BFM571 to KB cells in the absence and presence of P. gingivalis OMVs (Pgv). Attachment levels (means ± standard errors) were expressed as the percentage of attached bacteria relative to the total number

    Journal:

    Article Title: Porphyromonas gingivalis Vesicles Enhance Attachment, and the Leucine-Rich Repeat BspA Protein Is Required for Invasion of Epithelial Cells by "Tannerella forsythia"

    doi: 10.1128/IAI.00062-06

    Figure Lengend Snippet: Attachment of T. forsythia 43037 (Tf) and BFM571 to KB cells in the absence and presence of P. gingivalis OMVs (Pgv). Attachment levels (means ± standard errors) were expressed as the percentage of attached bacteria relative to the total number

    Article Snippet: In the presence of P. gingivalis OMVs, attachment of T. forsythia ATCC 43037 was increased fourfold.

    Techniques:

    Coaggregation of T. forsythia in the presence of P. gingivalis and outer membrane vesicles purified from P. gingivalis . Data are representative of three independent experiments. Data points represent means ± standard errors of triplicate readings.

    Journal:

    Article Title: Porphyromonas gingivalis Vesicles Enhance Attachment, and the Leucine-Rich Repeat BspA Protein Is Required for Invasion of Epithelial Cells by "Tannerella forsythia"

    doi: 10.1128/IAI.00062-06

    Figure Lengend Snippet: Coaggregation of T. forsythia in the presence of P. gingivalis and outer membrane vesicles purified from P. gingivalis . Data are representative of three independent experiments. Data points represent means ± standard errors of triplicate readings.

    Article Snippet: In the presence of P. gingivalis OMVs, attachment of T. forsythia ATCC 43037 was increased fourfold.

    Techniques: Purification