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Cytokine Profiling of Macrophages Exposed to Porphyromonas gingivalis, Its Lipopolysaccharide, or Its FimA Protein  

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To characterize the roles of Porphyromonas gingivalis and its components in the disease processes, we investigated the cytokine profile induced by live P. gingivalis, its lipopolysaccharides (LPS), and its major fimbrial protein, fimbrillin (FimA). Using cytokine antibody arrays, we found that P. gi...

Zhou, Qingde; Desta, Tesfahun; Fenton, Matthew; Graves, Dana T.; Amar, Salomon

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Effects of aging on endotoxin tolerance induced by lipopolysaccharides derived from Porphyromonas gingivalis and Escherichia coli.  

UK PubMed Central (United Kingdom)

BACKGROUND: Periodontitis is a bacterially induced chronic inflammatory disease. Exposure of the host to periodontal pathogens and their virulence factors induces a state of hyporesponsiveness to subsequent stimulations, termed endotoxin tolerance. Aging has a profound effect on immune response to bacteria challenge. The aim of this study was to explore the effects of aging on endotoxin tolerance induced by Porphyromonas gingivalis (P. gingivalis) lipopolysaccharide (LPS) and Escherichia coli (E. coli) LPS in murine peritoneal macrophages. METHODOLOGY/PRINCIPAL FINDINGS: We studied the cytokine production (TNF-? and IL-10) and Toll-like receptor 2, 4 (TLR2, 4) gene and protein expressions in peritoneal macrophages from young (2-month-old) and middle-aged (12-month-old) ICR mice following single or repeated P. gingivalis LPS or E. coli LPS stimulation. Pretreatment of peritoneal macrophages with P. gingivalis LPS or E. coli LPS resulted in a reduction in TNF-? production and an increase in IL-10 production upon secondary stimulation (p<0.05), and the markedly lower levels of TNF-? and higher levels of IL-10 were observed in macrophages from young mice compared with those from middle-aged mice (p<0.05). In addition, LPS restimulations also led to the significantly lower expression levels of TLR2, 4 mRNA and protein in macrophages from young mice (p<0.05). CONCLUSIONS/SIGNIFICANCE: Repeated LPS stimulations triggered endotoxin tolerance in peritoneal macrophages and the ability to develop tolerance in young mice was more excellent. The impaired ability to develop endotoxin tolerance resulted from aging might be related to TLR2, 4 and might lead to the incontrollable periodontal inflammation in older adults.

Sun Y; Li H; Yang MF; Shu W; Sun MJ; Xu Y

2012-01-01

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Porphyromonas gingivalis LIBRE DE POLISACÁRIDOS UTILIZANDO CROMATOGRAFÍA DE ALTA RESOLUCIÓN SEPHACRYL S-200 Purification of Porphyromonas gingivalis polysaccharide free lipopolysaccharide using Sephacryl S-200 high resolution chromatography  

Directory of Open Access Journals (Sweden)

Full Text Available El objetivo de este trabajo fue mejorar un método estándar para la purificación de lipopolisacárido (LPS) de Porphyromonas gingivalis libre de polisacáridos usando una estrategia de extracción, digestión enzimática y cromatografía de alta resolución. La bacteria P. gingivalis se cultivó en condiciones de anaerobiosis y se hizo extracción de las membranas con el método de fenol-agua. Luego de una digestión enzimática (DNAsa, RNAsa y proteasa) se separó el extracto por filtración por gel con Sephacryl S-200. La muestra purificada se caracterizó por electroforesis en gel de acrilamida con tinción de plata y por el método Purpald se detecto el ácido 2-ceto-3-desoxioctu-losónico (KDO). Se obtuvo una preparación libre de ácidos nucleicos, proteínas y polisacáridos. La separación por cromatografía fue de alta resolución al permitir la obtención de dos picos con diferentes componentes. El protocolo de purificación nos permitió obtener LPS de P. gingivalis con alto grado de pureza, el cual podría ser usado en próximos ensayos para evaluar su función en ensayos in vitro e in vivo; así como iniciar la obtención de LPS de otras bacterias periodontopáticas, con el fin de investigar la asociación de enfermedad periodontal con enfermedades cardiovasculares.The aim of this work was to improve a standard methodology to purify Porphyromonas gingivalis lipopolysaccharide (LPS) using a protocol of extraction, enzymatic digestion and high resolution chromatography. P. gingivalis bacteria was cultured in anaerobiosis, their membranes were extracted using the phenol-water method, then subjected to DNAse, RNAse and protease digestion and finally, the extract was separated by chromatography using Sephacryl S-200. The purified extract was characterized by silver staining after polyacrylamide gel electrophoresis and 2-keto-3-deoxioctanoic acid (KDO) was detected using the Purpald’s method. A preparation free of nucleic acid-, protein-or polysaccharides was obtained. The chromatographic separation showed high resolution since there was two discrete peaks with different components. The purification protocol allowed us to obtain a highly purified P. gingivalis LPS which could be used in future tests to evaluate its behavior in vitro and in vivo and elucidate its function, as well as to obtain LPS from other periodontopatic bacteria to address the association of periodontal disease with cardiovascular diseases.

DIEGO GUALTERO; JAIME E CASTELLANOS; GERARDO PÉREZ; GLORIA I LAFAURIE

2008-01-01

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Porphyromonas gingivalis LIBRE DE POLISACÁRIDOS UTILIZANDO CROMATOGRAFÍA DE ALTA RESOLUCIÓN SEPHACRYL S-200/ Purification of Porphyromonas gingivalis polysaccharide free lipopolysaccharide using Sephacryl S-200 high resolution chromatography  

Scientific Electronic Library Online (English)

Full Text Available Abstract in spanish El objetivo de este trabajo fue mejorar un método estándar para la purificación de lipopolisacárido (LPS) de Porphyromonas gingivalis libre de polisacáridos usando una estrategia de extracción, digestión enzimática y cromatografía de alta resolución. La bacteria P. gingivalis se cultivó en condiciones de anaerobiosis y se hizo extracción de las membranas con el método de fenol-agua. Luego de una digestión enzimática (DNAsa, RNAsa y proteasa) se separó el e (more) xtracto por filtración por gel con Sephacryl S-200. La muestra purificada se caracterizó por electroforesis en gel de acrilamida con tinción de plata y por el método Purpald se detecto el ácido 2-ceto-3-desoxioctu-losónico (KDO). Se obtuvo una preparación libre de ácidos nucleicos, proteínas y polisacáridos. La separación por cromatografía fue de alta resolución al permitir la obtención de dos picos con diferentes componentes. El protocolo de purificación nos permitió obtener LPS de P. gingivalis con alto grado de pureza, el cual podría ser usado en próximos ensayos para evaluar su función en ensayos in vitro e in vivo; así como iniciar la obtención de LPS de otras bacterias periodontopáticas, con el fin de investigar la asociación de enfermedad periodontal con enfermedades cardiovasculares. Abstract in english The aim of this work was to improve a standard methodology to purify Porphyromonas gingivalis lipopolysaccharide (LPS) using a protocol of extraction, enzymatic digestion and high resolution chromatography. P. gingivalis bacteria was cultured in anaerobiosis, their membranes were extracted using the phenol-water method, then subjected to DNAse, RNAse and protease digestion and finally, the extract was separated by chromatography using Sephacryl S-200. The purified extract (more) was characterized by silver staining after polyacrylamide gel electrophoresis and 2-keto-3-deoxioctanoic acid (KDO) was detected using the Purpald?s method. A preparation free of nucleic acid-, protein-or polysaccharides was obtained. The chromatographic separation showed high resolution since there was two discrete peaks with different components. The purification protocol allowed us to obtain a highly purified P. gingivalis LPS which could be used in future tests to evaluate its behavior in vitro and in vivo and elucidate its function, as well as to obtain LPS from other periodontopatic bacteria to address the association of periodontal disease with cardiovascular diseases.

GUALTERO, DIEGO; CASTELLANOS, JAIME E; PÉREZ, GERARDO; LAFAURIE, GLORIA I

2008-12-01

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Glycated matrix up-regulates inflammatory signaling similarly to Porphyromonas gingivalis lipopolysaccharide.  

UK PubMed Central (United Kingdom)

BACKGROUND AND OBJECTIVE: Hyperglycemia and advanced glycation end-products (AGEs) have been hypothesized as the etiologic factors of diabetic periodontitis. The aim of this study was to clarify in greater detail the patterns of AGE-mediated periodontal inflammation under various physiological conditions. MATERIAL AND METHODS: The deposition of AGEs and expression of the receptor for AGEs (RAGE) were identified by immunohistochemistry in Sprague-Dawley rats with experimentally induced periodontitis or diabetes. Human periodontal ligament cells (PDLCs) and mesenchymal stem cells (MSCs) were cultured under simulated conditions of hyperglycemia, Porphyromonas gingivalis lipopolysaccharide (LPS) stimulation and matrix glycation. Cell viability and expression of toll-like receptors (TLRs), Rage, an inflammatory signaling initiator (nuclear factor kappa light chain enhancer of activator ? cells), an oxidative stressor (heme oxygenase-1) and collagen synthesis (type I and type IV) genes were evaluated. RESULTS: The deposition of AGEs and the expression of Rage were evident in the inflamed periodontal tissues in all rats and appeared to be enhanced in rats with diabetes. Matrix glycation augmented cytotoxicity, up-regulated RAGE and TLRs in both PDLCs and MSCs, and significantly activated downstream inflammatory signaling in MSCs. Oxidative stress was significantly increased under matrix glycation in both PDLCs and MSCs and was significantly increased at a high-glucose concentration in MSCs. A consistent decrease in expression of type I and type IV collagens was observed in MSCs, but a delayed reduction was noted in PDLCs. CONCLUSIONS: Matrix glycation modulated cell behavior to induce inflammation equivalent to that produced by incubation with P. gingivalis LPS. Periodontal inflammation also led to matrix glycation, thus demonstrating a definite interaction between diabetes and periodontitis.

Chang PC; Chien LY; Chong LY; Kuo YP; Hsiao JK

2013-04-01

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Porphyromonas gingivalis Lipopolysaccharide Activates Canonical Wnt/?-Catenin and p38 MAPK Signalling in Stem Cells from the Apical Papilla.  

UK PubMed Central (United Kingdom)

As dental precursor cells, stem cells from the apical papilla (SCAP) are capable of forming roots and undergoing apexogenesis, which are impaired upon exposure to bacterial infection. Porphyromonas gingivalis is a common Gram-negative bacterium that is involved in pulpal and periapical infection. The purpose of this study was to investigate the effects of P. gingivalis lipopolysaccharide (LPS) on the Wnt/?-catenin and p38 mitogen-activated protein kinase (MAPK) signalling pathways in SCAP. As indicated by the IL-1? and TNF-? mRNA levels, P. gingivalis LPS induced the expression of pro-inflammatory cytokines in a dose-dependent manner. In addition, activation of the p38 MAPK and Wnt/?-catenin pathways was confirmed by the augmentation of phospho-p38 and ?-catenin protein expression and increased expression of c-myc and cyclin D1 mRNA. Despite no significant increase in ?-catenin mRNA expression, increased phosphorylation of glycogen synthase kinase (GSK)-3? suggested that GSK-3? was responsible for the accumulation of ?-catenin in the cytoplasm and translocation to the nucleus. Previous studies have shown that GSK-3? plays a critical role in crosstalk between the Wnt/?-catenin and p38 MAPK pathways. In the present study, we showed that the level of p38 phosphorylation decreased upon pretreatment with a p38 MAPK inhibitor for 1 h before stimulating SCAP with 10 ?g/ml P. gingivalis LPS. However, the levels of GSK-3? and ?-catenin phosphorylation in the cytoplasm and nucleus were not significantly altered. Our results suggest that the p38 MAPK and canonical Wnt/?-catenin signalling pathways are activated by P. gingivalis LPS in SCAP, but we have no evidence that p38 MAPK is upstream of GSK-3? in the Wnt/?-catenin signalling pathway.

Wang J; Dai J; Liu B; Gu S; Cheng L; Liang J

2013-07-01

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The expression and regulation of matrix metalloproteinase-3 is critically modulated by Porphyromonas gingivalis lipopolysaccharide with heterogeneous lipid A structures in human gingival fibroblasts.  

UK PubMed Central (United Kingdom)

BACKGROUND: Porphyromonas gingivalis lipopolysaccharide (LPS) is a crucial virulence factor strongly associated with chronic periodontitis which is the primary cause of tooth loss in adults. It exhibits remarkable heterogeneity containing tetra-(LPS(1435/1449)) and penta-(LPS(1690)) acylated lipid A structures. Human gingival fibroblasts (HGFs) as the main resident cells of human gingiva play a key role in regulating matrix metalloproteinases (MMPs) and contribute to periodontal homeostasis. This study investigated the expression and regulation of MMPs1-3 and tissue inhibitors of MMP-1 (TIMP-1) in HGFs in response to P. gingivalis LPS(1435/1449) and LPS(1690) and hexa-acylated E. coli LPS as a reference. The expression of MMPs 1-3 and TIMP-1 was evaluated by real-time PCR and ELISA. RESULTS: The MMP-3 mRNA and protein were highly upregulated in P. gingivalis LPS(1690)- and E. coli LPS-treated cells, whereas no induction was observed in P. gingivalis LPS(1435/1449)-treated cells. On the contrary, the expression of MMP-1 and -2 was not significantly affected by P. gingivalis LPS lipid A heterogeneity. The TIMP-1 mRNA was upregulated in P. gingivalis LPS(1435/1449)- and E. coli LPS-treated cells. Next, signal transduction pathways involved in P. gingivalis LPS-induced expression of MMP-3 were examined by blocking assays. Blockage of p38 MAPK and ERK significantly inhibited P. gingivalis LPS(1690)-induced MMP-3 expression in HGFs. CONCLUSION: The present findings suggest that the heterogeneous lipid A structures of P. gingivalis LPS differentially modulate the expression of MMP-3 in HGFs, which may play a role in periodontal pathogenesis.

Herath TD; Wang Y; Seneviratne CJ; Darveau RP; Wang CY; Jin L

2013-01-01

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Whole Blood Cultures From Patients With Chronic Periodontitis Respond Differently to Porphyromonas gingivalis but not Escherichia coli Lipopolysaccharide.  

UK PubMed Central (United Kingdom)

Background: Porphyromonas gingivalis lipid A heterogeneity modulates cytokine expression in human cells. This study investigated the effects of two lipid A isoforms of P. gingivalis, LPS1435/1449 and LPS1690, on the secretion of pro-inflammatory and regulatory cytokines in total blood cultures from patients with and without chronic periodontitis. Methods: A cross-sectional study was conducted in thirty-eight systemically healthy individuals divided in two groups: chronic periodontitis (CP) group (n=19)- patients diagnosed with chronic periodontitis, and no periodontitis (NP) group (n=19)- control patients without periodontitis. Blood samples were collected from all patients and whole blood cell cultures (WBCC) were stimulated for 48h with P. gingivalis LPS1435/1449, LPS1690 and E. coli LPS. Unstimulated WBCC served as negative controls. The secretion of IFN?, IL-10 and TGF? was detected in WBCC supernatants by enzyme-linked immunosorbent assays. Results: E. coli LPS significantly increased the expression of all cytokines in WBCC from both NP and CP groups when compared to non-stimulated cells (control treatment). P. gingivalis LPS preparations increased IFN? levels in CP group but not in NP group when compared to control (P<0.05). P. gingivalis LPS preparations also increased IL-10 and TGF? levels in both CP and NP groups, however P. gingivalis LPS1690 showed a 3-fold increase on IL-10 production in NP group (P<0.05) when compared with P. gingivalis LPS1435/144. Conclusions: These data demonstrate that WBCC cell populations obtained from healthy and chronic periodontitis patients may differ in the cytokine response to P. gingivalis but not E. coli LPS. This is consistent with the notion that chronic periodontitis alters the systemic WBCC response and that this can be detected by the different P. gingivalis LPS structures.

Nogueira-Filho G; Rosa BT; Santos PF; Tunes UR; Freire SM; Meyer R; Darveau RP

2013-09-01

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Resistin in gingival crevicular fluid and induction of resistin release by Porphyromonas gingivalis lipopolysaccharide in human neutrophils.  

UK PubMed Central (United Kingdom)

BACKGROUND AND OBJECTIVE: Resistin is an adipocytokine that induces insulin resistance and is predominantly expressed in adipocytes and peripheral blood mononuclear cells. Resistin expression increases in inflammatory diseases as well as diabetes mellitus, and is upregulated by bacterial pathogens and proinflammatory cytokines. The aim of this study was to identify resistin in human gingival crevicular fluid, to compare the resistin levels in gingival crevicular fluid between subjects with and without periodontitis and diabetes mellitus and to investigate the regulation of resistin release from human neutrophils by Porphyromonas gingivalis lipopolysaccharide (P-LPS). MATERIAL AND METHODS: Gingival crevicular fluid samples were collected from patients with chronic periodontitis (n = 24), patients with diabetes mellitus-related periodontitis (n = 18) and healthy subjects (n = 21). Resistin in gingival crevicular fluid was determined using western blot analysis and an ELISA kit. The glycated hemoglobin (HbA(1c)) value was obtained from patients with diabetes mellitus-related periodontitis by a medical interview. Human neutrophils were cultured with P-LPS (0-1000 ng/mL), or incubated with inhibitors of actin or microtubule polymerization in the absence or presence of P-LPS. The medium and cellular fractions were used for determination of resistin by ELISA. RESULTS: The resistin level in gingival crevicular fluid from patients with periodontitis or diabetes mellitus-related periodontitis was significantly higher than that of healthy subjects. The resistin level in gingival crevicular fluid was correlated with gingival index score, but not blood HbA(1c) value. The P-LPS increased resistin release from human neutrophils, and its induction was decreased by actin polymerization inhibitors. CONCLUSION: We show, for the first time, the presence of resistin in gingival crevicular fluid. A high resistin level in gingival crevicular fluid samples from periodontitis patients may to some extent be related to P-LPS-induced resistin release from neutrophils.

Hiroshima Y; Bando M; Inagaki Y; Mihara C; Kataoka M; Murata H; Shinohara Y; Nagata T; Kido J

2012-10-01

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Endotoxin Tolerance Induced by Lipopolysaccharides Derived from Porphyromonas gingivalis and Escherichia coli: Alternations in Toll-Like Receptor 2 and 4 Signaling Pathway.  

UK PubMed Central (United Kingdom)

Periodontitis is a chronic inflammatory disease induced by bacteria. Exposure of the host to periodontal pathogens and their virulence factors induces a hyporesponsive state to subsequent challenge, which is termed endotoxin tolerance. In this experiment, we studied the cytokine production in THP-1 cells upon single or repeated Porphyromonas gingivalis (P. gingivalis) lipopolysaccharide (LPS) or Escherichia coli (E. coli) LPS stimulation by ELISA. In addition, the protein expression profiles of Toll-like receptor 2 (TLR2), TLR4, IL-1 receptor-associated kinase 4 (IRAK4) and IRAK-M and the gene expression changes of Toll-interacting protein (Tollip) and suppressor of cytokine-signaling-1 (SOCS1) were explored to identify possible mechanisms for changes in cytokine secretion. After repeated stimulation with P. gingivalis LPS or E. coli LPS, secretions of TNF-? and IL-1? were decreased significantly compared with those following single challenge, while the levels of IL-10 were increased (p?gingivalis LPS-tolerized cells (p?>?0.05). In addition, severe downregulation of TLR2 was detected in THP-1 cells retreated with P. gingivalis LPS, and the reduction of TLR4 expression was observed in cells restimulated with E. coli LPS (p?gingivalis LPS or E. coli LPS also led to an enhancement of IRAK-M and SOCS1, while maintaining the expressions of IRAK4 and Tollip. This pattern of cytokine production indicates the different effects of endotoxin tolerance triggered by P. gingivalis LPS and E. coli LPS, which might contribute to limiting inflammatory damage. Moreover, TLR2, TLR4, IRAK-M, and SOCS1 might play important roles in developing tolerance.

Sun Y; Li H; Sun MJ; Zheng YY; Gong DJ; Xu Y

2013-10-01

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Porphyromonas gingivalis endotoxin affinity for dental ceramics.  

UK PubMed Central (United Kingdom)

This study evaluated the effects of chemical composition, surface treatment, and initial exposure dose on Porphyromonas gingivalis lipopolysaccharide adherence to and elution from dental ceramics. Lipopolysaccharide, commonly known as endotoxin, can initiate a variety of biologic responses. Opaque, body, and Dicor ceramic disks were individually exposed to 250, 1000, or 2500 EU/ml 3H-lipopolysaccharide and incubated for 24 hours at 37 degrees C. Disks were then transferred to fresh lipopolysaccharide-free water and incubated for up to 96 hours to evaluate elution. Mean initial lipopolysaccharide adherence ranged from 0.397 +/- 0.048 EU/mm2 to 5.056 +/- 0.117 EU/mm2. Greater initial exposure levels resulted in greater adherence, and at higher lipopolysaccharide exposure levels, lipopolysaccharide adherence differences were based on ceramic type. Mean lipopolysaccharide elution levels ranged from 0.063 +/- 0.02 EU/mm2 to 0.00 EU/mm2 at 96 hours for all groups. Greater initial adherence resulted in greater elution. Ceramic type did not affect elution. Surface finish affected elution at the 2500 EU exposure level. The affinity of lipopolysaccharide for dental ceramics could contribute to a periodontal inflammatory process.

Robinson FG; Knoernschild KL; Sterrett JD; Tompkins GR

1996-02-01

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Porphyromonas gingivalis: a clonal pathogen?  

Directory of Open Access Journals (Sweden)

Full Text Available The introduction of multilocus sequence typing (MLST) in infectious disease research has allowed standardized typing of bacterial clones. Through multiple markers around the genome, it is possible to determine the sequence type (ST) of bacterial isolates to establish the population structure of a species. For the periodontal pathogen, Porphyromonas gingivalis, the MLST scheme has been established at www.pubmlst.org/pgingivalis, and data from the database indicate a high degree of genetic diversity and a weakly clonal population structure comparable with Neisseria menigitidis. The major fimbriae (FimA) have been held responsible for the adhesive properties of P. gingivalis and represent an important virulence factor. The fimA genotyping method (PCR based) indicate that fimA genotype II, IV and Ib are associated with diseased sites in periodontitis and tissue specimens from cardiovascular disease. fimA genotyping of the isolates in the MLST database supports the association of genotypes II and IV with periodontitis. As a result of multiple positive PCR reactions in the fimA genotyping, sequencing of the fimA gene revealed only minor nucleotide variation between isolates of the same and different genotypes, suggesting that the method should be redesigned or re-evaluated. Results from several investigations indicate a higher intraindividual heterogeneity of P. gingivalis than found earlier. Detection of multiple STs from one site in several patients with “refractory” periodontitis, showed allelic variation in two housekeeping genes indicating recombination between different clones within the periodontal pocket.

Morten Enersen

2011-01-01

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Lipopolysaccharide preparation extracted from Porphyromonas gingivalis lipoprotein-deficient mutant shows a marked decrease in toll-like receptor 2-mediated signaling.  

Science.gov (United States)

We recently demonstrated that a new PG1828-encoded lipoprotein (PG1828LP) was able to be separated from a Porphyromonas gingivalis lipopolysaccharide (LPS) preparation, and we found that it exhibited strong cell activation, similar to that of Escherichia coli LPS, through a Toll-like receptor 2 (TLR2)-dependent pathway. In order to determine the virulence of PG1828LP toward cell activation, we generated a PG1828-deficient mutant of P. gingivalis strain 381 by allelic exchange mutagenesis using an ermF-ermAM antibiotic resistance cassette. A highly purified preparation of LPS from a PG1828-deficient mutant (DeltaPG1828-LPS) showed nearly the same ladder-like patterns in silver-stained gels as a preparation of LPS from a wild-type strain (WT-LPS), as well as Limulus amoebocyte lysate activities that were similar to those of the WT-LPS preparation. However, the ability of the DeltaPG1828-LPS preparation to activate NF-kappaB in TLR2-expressing cells was markedly attenuated. Cytokine production by human gingival fibroblasts was also decreased in response to the DeltaPG1828-LPS preparation in comparison with the WT-LPS preparation, and the activity was comparable to the stimulation of highly purified lipid A of P. gingivalis by TLR4. Further, lethal toxicity was rarely observed following intraperitoneal injection of the PG1828-deficient mutant into mice compared to that with the wild-type strain, while the DeltaPG1828-LPS preparation showed no lethal toxicity. Taken together, these results clearly indicate that PG1828LP plays an essential role in inflammatory responses and may be a major virulence factor of P. gingivalis. PMID:15784558

Asai, Yasuyuki; Hashimoto, Masahito; Fletcher, Hansel M; Miyake, Kensuke; Akira, Shizuo; Ogawa, Tomohiko

2005-04-01

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Lipopolysaccharide preparation extracted from Porphyromonas gingivalis lipoprotein-deficient mutant shows a marked decrease in toll-like receptor 2-mediated signaling.  

UK PubMed Central (United Kingdom)

We recently demonstrated that a new PG1828-encoded lipoprotein (PG1828LP) was able to be separated from a Porphyromonas gingivalis lipopolysaccharide (LPS) preparation, and we found that it exhibited strong cell activation, similar to that of Escherichia coli LPS, through a Toll-like receptor 2 (TLR2)-dependent pathway. In order to determine the virulence of PG1828LP toward cell activation, we generated a PG1828-deficient mutant of P. gingivalis strain 381 by allelic exchange mutagenesis using an ermF-ermAM antibiotic resistance cassette. A highly purified preparation of LPS from a PG1828-deficient mutant (DeltaPG1828-LPS) showed nearly the same ladder-like patterns in silver-stained gels as a preparation of LPS from a wild-type strain (WT-LPS), as well as Limulus amoebocyte lysate activities that were similar to those of the WT-LPS preparation. However, the ability of the DeltaPG1828-LPS preparation to activate NF-kappaB in TLR2-expressing cells was markedly attenuated. Cytokine production by human gingival fibroblasts was also decreased in response to the DeltaPG1828-LPS preparation in comparison with the WT-LPS preparation, and the activity was comparable to the stimulation of highly purified lipid A of P. gingivalis by TLR4. Further, lethal toxicity was rarely observed following intraperitoneal injection of the PG1828-deficient mutant into mice compared to that with the wild-type strain, while the DeltaPG1828-LPS preparation showed no lethal toxicity. Taken together, these results clearly indicate that PG1828LP plays an essential role in inflammatory responses and may be a major virulence factor of P. gingivalis.

Asai Y; Hashimoto M; Fletcher HM; Miyake K; Akira S; Ogawa T

2005-04-01

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Gingipain aminopeptidase activities in Porphyromonas gingivalis.  

UK PubMed Central (United Kingdom)

Bestatin, a specific inhibitor of metalloaminopeptidases,inhibits the growth of Porphyromonas gingivalis. To identify its target enzyme, a library of fluorescent substrates was used but no metalloaminopeptidase activity was found. The aminopeptidase activity of P. gingivalis was bestatin-insensitive and directed exclusively toward N-terminal arginine and lysine substrates. Class-specific inhibitors and gingipain-null mutants showed that gingipains were the only enzymes responsible for this activity.The kinetic constants obtained for Rgps were comparable to those of human aminopeptidases but Kgp aminopeptidase activity was weaker. This finding reveals a new role for gingipains as aminopeptidases in the degradation of proteins and peptides in P. gingivalis.

Veillard F; Potempa B; Poreba M; Drag M; Potempa J

2012-12-01

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Induction of toll-like receptor expression by Porphyromonas gingivalis.  

UK PubMed Central (United Kingdom)

BACKGROUND: Toll-like receptors (TLRs) play pivotal roles in host immune responses and have been suggested to be involved in the development of many infectious diseases. In this study, the mRNA expression levels of TLR2, TLR4, and TLR9 and their relationship with periodontopathic bacteria in periodontal tissue are examined. Furthermore, the mechanism of TLR induction by Porphyromonas gingivalis is investigated in human gingival fibroblasts (HGFs). METHODS: Gingival tissue and subgingival plaque samples were collected from 19 patients with chronic periodontitis (CP) and 16 control individuals without periodontitis. Gene expression levels in the tissues and in HGFs were analyzed by reverse transcription-polymerase chain reaction (RT-PCR). The numbers of periodontopathic bacteria were determined by quantitative real-time PCR. RESULTS: The expression levels of TLR2 and TLR9 were significantly higher in the tissues of patients with CP compared to the tissues of control individuals. The mRNA levels of TLR2 and TLR9, but not TLR4, were positively correlated with the number of P. gingivalis in subgingival plaque. P. gingivalis sonicated extract, P. gingivalis lipopolysaccharide, P. gingivalis DNA, and tumor necrosis factor-?(TNF-?) could significantly upregulate the mRNA expression of TLR2 in HGFs. Furthermore, P. gingivalis-mediated TLR2 expression was suppressed by TNF-? antibody. CONCLUSIONS: This study suggests that P. gingivalis infection induces TLR2 and TLR9 upregulation in patients with CP. P. gingivalis-induced TLR2 expression in HGFs is partially dependent on TNF-? and may lead to sensitization of HGFs to bacterial components encountered in the periodontal microenvironment.

Wara-aswapati N; Chayasadom A; Surarit R; Pitiphat W; Boch JA; Nagasawa T; Ishikawa I; Izumi Y

2013-07-01

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Porphyromonas gingivalis biofilms persist after chlorhexidine treatment.  

UK PubMed Central (United Kingdom)

Chlorhexidine (CHX) gluconate effectively reduces the viability of biofilm-forming bacteria, such as Porphyromonas gingivalis. However, it is impossible to completely remove biofilms. The goal of the present study was to assess the potential pathogenicity of residual P. gingivalis biofilms in vitro after treatment with CHX gluconate. Scanning and transmission electron microscopy and confocal laser imaging revealed that treatment with CHX gluconate disrupted individual biofilm-forming P. gingivalis cells but did not destroy the biofilms. The volumes of the protein and carbohydrate constituents in the residual biofilms were not significantly different from those of the controls. The physical resistance of the residual biofilms to ultrasonication was significantly higher than that of controls. The volume of P. gingivalis adherent to the residual biofilms was higher than that to saliva-coated wells. These findings suggest that although CHX gluconate caused disruption of biofilm-forming cells, the constituents derived from disrupted cells were maintained in the biofilms, which sustained their external structures. Moreover, the residual biofilms could serve as a scaffold for the formation of new biofilms.

Yamaguchi M; Noiri Y; Kuboniwa M; Yamamoto R; Asahi Y; Maezono H; Hayashi M; Ebisu S

2013-06-01

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Identification of Proteins Differentially Expressed in Human Monocytes Exposed to Porphyromonas gingivalis and Its Purified Components by High-Throughput Immunoblotting  

Digital Repository Infrastructure Vision for European Research (DRIVER)

To characterize the roles of Porphyromonas gingivalis and its components in disease processes, we investigated the cytokine profiles induced by live P. gingivalis, its lipopolysaccharide (LPS), and its major fimbrial protein, fimbrillin (FimA). A cytokine antibody array revealed that human monocyte-...

Zhou, Qingde; Amar, Salomon

19

Inhibitory effects of tocopherols on expression of the cyclooxygenase-2 gene in RAW264.7 cells stimulated by lipopolysaccharide, tumor necrosis factor-? or Porphyromonas gingivalis fimbriae.  

UK PubMed Central (United Kingdom)

BACKGROUND: Tocopherols, which include ?-, ?-, ?-, and ?-tocopherol, protect cells against harmful free radicals and play an important role in preventing many human diseases such as cancer, inflammatory disorders, and ageing itself. However, the causal relationships between periodontal or oral chronic diseases and tocopherols have not been sufficiently studied. The present study investigated the inhibitory effects of these compounds on the expression of cyclooxygenase-2 (COX2) mRNA in RAW264.7 cells stimulated with lipopolysaccharide (LPS), tumor necrosis factor-? (TNF?) or fimbriae of Poryphyromonas gingivalis (Pg), an oral anaerobe. MATERIALS AND METHODS: The cytotoxicity (EC??) of tocopherols toward RAW cells was determined using a cell counting kit (CCK-8). The regulatory effect of these compounds on the expression of COX2 mRNA stimulated with LPS, TNF? or Pg fimbriae was investigated using real-time polymerase chain reaction (PCR). RESULTS: Each tocopherol had similarly low cytotoxicity. COX2 gene expression in RAW cells after exposure to the three different macrophage activators was inhibited by the tocopherols (p<0.01). Compared to ?-tocopherol, ?-, ?- and ?-tocopherol exhibited greater inhibitory effects (p<0.05). CONCLUSION: Tocopherols exhibit anti-inflammatory activity, and ?-, ?- and ?-tocopherol have particularly more potent anti-inflammatory activity than ?-tocopherol. Tocopherols may have potential utility for prevention of periodontal and chronic oral diseases.

Murakami Y; Kawata A; Koh T; Seki Y; Tamura S; Katayama T; Fujisawa S

2013-07-01

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Designing a peptide-based vaccine against Porphyromonas gingivalis.  

UK PubMed Central (United Kingdom)

Using proteome databases and exploiting the concept that a rare sequence is a potential epitope, epitopic sequences derived from Porphyromonas gingivalis fimA type I protein were examined for pentapeptide sequence similarity score to the human proteome. We obtained data showing that most of the linear bacterial determinants are (or are formed by) peptide fragment(s) absent (or rarely found) in the human proteins. These results seem to confirm the hypothesis that low-sequence similarity may contribute to shape the epitope repertoire and provides a potential tool for designing new immunotherapeutic approaches to apply in Porphyromonas gingivalis infected periodontitis.

Lucchese A; Guida A; Capone G; Petruzzi M; Lauritano D; Serpico R

2013-01-01

 
 
 
 
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Designing a peptide-based vaccine against Porphyromonas gingivalis.  

Science.gov (United States)

Using proteome databases and exploiting the concept that a rare sequence is a potential epitope, epitopic sequences derived from Porphyromonas gingivalis fimA type I protein were examined for pentapeptide sequence similarity score to the human proteome. We obtained data showing that most of the linear bacterial determinants are (or are formed by) peptide fragment(s) absent (or rarely found) in the human proteins. These results seem to confirm the hypothesis that low-sequence similarity may contribute to shape the epitope repertoire and provides a potential tool for designing new immunotherapeutic approaches to apply in Porphyromonas gingivalis infected periodontitis. PMID:23277074

Lucchese, Alberta; Guida, Agostino; Capone, Giovanni; Petruzzi, Massimo; Lauritano, Dorina; Serpico, Rosario

2013-01-01

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Nasal Immunization with Porphyromonas gingivalis Outer Membrane Protein Decreases P. gingivalis-Induced Atherosclerosis and Inflammation in Spontaneously Hyperlipidemic Mice?  

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Porphyromonas gingivalis has been shown to accelerate atherosclerotic lesion development in hyperlipidemic animals. We assessed the potential of a nasal vaccine against P. gingivalis infection for the prevention of atherosclerosis. Apolipoprotein E-deficient spontaneously hyperlipidemic (Apoeshl) mi...

Koizumi, Y.; Kurita-Ochiai, T.; Oguchi, S.; Yamamoto, M.

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Porphyromonas gingivalis decreases osteoblast proliferation through IL-6-RANKL/OPG and MMP-9/TIMPs pathways  

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Full Text Available Background: Porphyromonas gingivalis, an important periodontal pathogen, is closely associated with inflammatory alveolar bone resorption. This bacterium exerts its pathogenic effect indirectly through multiple virulence factors, such as lipopolysaccharides, fimbriae, and proteases. Another possible pathogenic path may be through a direct interaction with the host?s soft and hard tissues (e.g., alveolar bone), which could lead to periodontitis. Aims and Objectives: The aim of the present study was to investigate the direct effect of live and heat-inactivated P gingivalis on bone resorption, using an in vitro osteoblast culture model. Results: Optical microscopy and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide MTT assay revealed that live P gingivalis induced osteoblast detachment and reduced their proliferation. This effect was specific to live bacteria and was dependent on their concentration. Live P gingivalis increased IL-6 mRNA expression and protein production and downregulated RANKL and OPG mRNA expression. The effect of live P gingivalis on bone resorption was strengthened by an increase in MMP-9 expression and its activity. This increase was accompanied by an increase in TIMP-1 and TIMP-2 mRNA expression and protein production by osteoblasts infected with live P gingivalis. Conclusion: Overall, the results suggest that direct contact of P gingivalis with osteoblasts induces bone resorption through an inflammatory pathway that involves IL-6, RANKL/OPG, and MMP-9/TIMPs.

Le Xuan; Laflamme Claude; Rouabhia Mahmoud

2009-01-01

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Outer membrane vesicles of Porphyromonas gingivalis elicit a mucosal immune response.  

UK PubMed Central (United Kingdom)

We previously reported that mutation of galE in Porphyromonas gingivalis has pleiotropic effects, including a truncated lipopolysaccharide (LPS) O-antigen and deglycosylation of the outer membrane protein OMP85 homolog. In the present study, further analysis of the galE mutant revealed that it produced little or no outer membrane vesicles (OMVs). Using three mouse antisera raised against whole cells of the P. gingivalis wild type strain, we performed ELISAs to examine the reactivity of these antisera with whole cells of the wild type or the galE mutant. All three antisera had significantly lower reactivity against the galE mutant compared to wild type. OMVs, but not LPS, retained the immunodominant determinant of P. gingivalis, as determined by ELISAs (with wild type LPS or OMVs as antigen) and absorption assays. In addition, we assessed the capacity of OMVs as a vaccine antigen by intranasal immunization to BALB/c mice. Synthetic double-stranded RNA polyriboinosinic polyribocytidylic acid [Poly (I?C)], an agonist of Toll-like receptor 3 (TLR3), was used as the mucosal adjuvant. Vaccination with OMV elicited dramatically high levels of P. gingivalis-specific IgA in nasal washes and saliva, as well as serum IgG and IgA. In conclusion, the OMVs of P. gingivalis have an important role in mucosal immunogenicity as well as in antigenicity. We propose that P. gingivalis OMV is an intriguing immunogen for development of a periodontal disease vaccine.

Nakao R; Hasegawa H; Ochiai K; Takashiba S; Ainai A; Ohnishi M; Watanabe H; Senpuku H

2011-01-01

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Porphyromonas gingivalis and Treponema denticola synergistic polymicrobial biofilm development.  

Science.gov (United States)

Chronic periodontitis has a polymicrobial biofilm aetiology and interactions between key bacterial species are strongly implicated as contributing to disease progression. Porphyromonas gingivalis, Treponema denticola and Tannerella forsythia have all been implicated as playing roles in disease progression. P. gingivalis cell-surface-located protease/adhesins, the gingipains, have been suggested to be involved in its interactions with several other bacterial species. The aims of this study were to determine polymicrobial biofilm formation by P. gingivalis, T. denticola and T. forsythia, as well as the role of P. gingivalis gingipains in biofilm formation by using a gingipain null triple mutant. To determine homotypic and polymicrobial biofilm formation a flow cell system was employed and the biofilms imaged and quantified by fluorescent in situ hybridization using DNA species-specific probes and confocal scanning laser microscopy imaging. Of the three species, only P. gingivalis and T. denticola formed mature, homotypic biofilms, and a strong synergy was observed between P. gingivalis and T. denticola in polymicrobial biofilm formation. This synergy was demonstrated by significant increases in biovolume, average biofilm thickness and maximum biofilm thickness of both species. In addition there was a morphological change of T. denticola in polymicrobial biofilms when compared with homotypic biofilms, suggesting reduced motility in homotypic biofilms. P. gingivalis gingipains were shown to play an essential role in synergistic polymicrobial biofilm formation with T. denticola. PMID:23990979

Zhu, Ying; Dashper, Stuart G; Chen, Yu-Yen; Crawford, Simon; Slakeski, Nada; Reynolds, Eric C

2013-08-26

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Identification of interspecies interactions affecting Porphyromonas gingivalis virulence phenotypes  

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Full Text Available Background: Periodontitis is recognized as a complex polymicrobial disease, however, the impact of the bacterial interactions among the 700–1,000 different species of the oral microbiota remains poorly understood. We conducted an in vitro screen for oral bacteria that mitigate selected virulence phenotypes of the important periodontal pathogen, Porphyromonas gingivalis. Methods: We isolated and identified oral anaerobic bacteria from subgingival plaque of dental patients. When cocultured with P. gingivalis W83, specific isolates reduced the cytopathogenic effects of P. gingivalis on oral epithelial cells. Results: In an initial screen of 103 subgingival isolates, we identified 19 distinct strains from nine species of bacteria (including Actinomyces naeslundii, Streptococcus oralis, Streptococcus mitis, and Veilonella dispar) that protect oral epithelial cells from P. gingivalis-induced cytotoxicity. We found that some of these strains inhibited P. gingivalis growth in plate assays through the production of organic acids, whereas some decreased the gingipain activity of P. gingivalis in coculture or mixing experiments. Conclusion: In summary, we identified 19 strains isolated from human subgingival plaque that interacted with P. gingivalis, resulting in mitigation of its cytotoxicity to oral epithelial cells, inhibition of growth, and/or reduction of gingipain activity. Understanding the mechanisms of interaction between bacteria in the oral microbial community may lead to the development of new probiotic agents and new strategies for interrupting the development of periodontal disease.

Elizabeth L. Tenorio; Brian A. Klein; Wai S. Cheung; Linden T. Hu

2011-01-01

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TRANSMISSION OF Porphyromonas gingivalis FROM CAREGIVERS TO CHILDREN.  

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Full Text Available Periodontal diseases are socially significant diseases, which occur in adults but in children and adolescents as well. Despite a low prevalence of aggressive periodontitis at a young age, its severity is a challenge for pediatric dentistry. The goal of this study is to find if the prevalence of Porphyromonas gingivalis among children whose parents suffer from periodontal diseases is greater than among children with healthy parents. Methods:- Polymerase chain reaction (PCR).- Culture method. When PCR was used P.gingivalis was found in 35.5% of parents with periodontitis and in 6,5% of their children, children with healthy parents and their parents. No statistically significant relation (P>0.05) between periodontal parents and their children was found. When culture method was used P.gingivalis was not detected.Studying such correlations and standardizing methods of detection could contribute the evaluation of periodontal disease risk in adolescents.

Marieta D. Belcheva; Angelina Kiselova-Yaneva; Maya Rashkova; Tsonko Uzunov; Assya Krasteva; Stefan Panaiotov; Victoria Levterova; Sashka Targova; Milena Georgieva; Nadia Brankova

2012-01-01

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Antibody response to a chromatographic fraction of Porphyromonas gingivalis and its correlation with periodontal status  

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Porphyromonas gingivalis tem sido fortemente associado à gravidade das doenças periodontais e estimula respostas celulares e humorais no hospedeiro. Objetivo: Correlacionar os níveis de IgA, IgG e subclasses de IgG contra uma fração cromatográfica do extrato de Porphyromonas gingivalis ATCC33277 ex...

Trindade, Soraya Castro et. al

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Identification of essential genes of the periodontal pathogen Porphyromonas gingivalis  

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Full Text Available Abstract Background Porphyromonas gingivalis is a Gram-negative anaerobic bacterium associated with periodontal disease onset and progression. Genetic tools for the manipulation of bacterial genomes allow for in-depth mechanistic studies of metabolism, physiology, interspecies and host-pathogen interactions. Analysis of the essential genes, protein-coding sequences necessary for survival of P. gingivalis by transposon mutagenesis has not previously been attempted due to the limitations of available transposon systems for the organism. We adapted a Mariner transposon system for mutagenesis of P. gingivalis and created an insertion mutant library. By analyzing the location of insertions using massively-parallel sequencing technology we used this mutant library to define genes essential for P. gingivalis survival under in vitro conditions. Results In mutagenesis experiments we identified 463 genes in P. gingivalis strain ATCC 33277 that are putatively essential for viability in vitro. Comparing the 463 P. gingivalis essential genes with previous essential gene studies, 364 of the 463 are homologues to essential genes in other species; 339 are shared with more than one other species. Twenty-five genes are known to be essential in P. gingivalis and B. thetaiotaomicron only. Significant enrichment of essential genes within Cluster of Orthologous Groups ‘D’ (cell division), ‘I’ (lipid transport and metabolism) and ‘J’ (translation/ribosome) were identified. Previously, the P. gingivalis core genome was shown to encode 1,476 proteins out of a possible 1,909; 434 of 463 essential genes are contained within the core genome. Thus, for the species P. gingivalis twenty-two, seventy-seven and twenty-three percent of the genome respectively are devoted to essential, core and accessory functions. Conclusions A Mariner transposon system can be adapted to create mutant libraries in P. gingivalis amenable to analysis by next-generation sequencing technologies. In silico analysis of genes essential for in vitro growth demonstrates that although the majority are homologous across bacterial species as a whole, species and strain-specific subsets are apparent. Understanding the putative essential genes of P. gingivalis will provide insights into metabolic pathways and niche adaptations as well as clinical therapeutic strategies.

Klein Brian A; Tenorio Elizabeth L; Lazinski David W; Camilli Andrew; Duncan Margaret J; Hu Linden T

2012-01-01

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Proteomics of Porphyromonas gingivalis within a model oral microbial community  

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Full Text Available Abstract Background Porphyromonas gingivalis is a periodontal pathogen that resides in a complex multispecies microbial biofilm community known as dental plaque. Confocal laser scanning microscopy showed that P. gingivalis can assemble into communities in vitro with Streptococcus gordonii and Fusobacterium nucleatum, common constituents of dental plaque. Whole cell quantitative proteomics, along with mutant construction and analysis, were conducted to investigate how P. gingivalis adapts to this three species community. Results 1156 P. gingivalis proteins were detected qualitatively during comparison of the three species model community with P. gingivalis incubated alone under the same conditions. Integration of spectral counting and summed signal intensity analyses of the dataset showed that 403 proteins were down-regulated and 89 proteins up-regulated. The proteomics results were inspected manually and an ontology analysis conducted using DAVID. Significant decreases were seen in proteins involved in cell shape and the formation of the cell envelope, as well as thiamine, cobalamin, and pyrimidine synthesis and DNA repair. An overall increase was seen in proteins involved in protein synthesis. HmuR, a TonB dependent outer membrane receptor, was up-regulated in the community and an hmuR deficient mutant was deficient in three species community formation, but was unimpaired in its ability to form mono- or dual-species biofilms. Conclusion Collectively, these results indicate that P. gingivalis can assemble into a heterotypic community with F. nucleatum and S. gordonii, and that a community lifestyle provides physiologic support for P. gingivalis. Proteins such as HmuR, that are up-regulated, can be necessary for community structure.

Kuboniwa Masae; Hendrickson Erik L; Xia Qiangwei; Wang Tiansong; Xie Hua; Hackett Murray; Lamont Richard J

2009-01-01

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Characterisation of the ?- and ?-Mannosidases of Porphyromonas gingivalis.  

UK PubMed Central (United Kingdom)

Mannose is an important sugar in the biology of the gram-negative bacterium, Porphyromonas gingivalis. It is a major component of the oligosaccharides attached to the cysteine-proteases Arg-gingipains, the repeating units of an acidic LPS (A-LPS) and the core-regions of both types of LPS produced by this organism (O-LPS and A-LPS) and a reported extra-cellular polysaccharide (EPS) isolated from spent culture medium. The organism occurs at inflamed sites in periodontal tissues where it is exposed to host glycoproteins rich in mannose which may be substrates for the acquisition of mannose by P. gingivalis. Five potential mannosidases were identified in the P. gingivalis W83 genome which may play a role in mannose acquisition. Four mannosidases were characterised in this study: PG0032 was a ?-mannosidase whereas PG0902 and PG1712 were capable of hydrolysing p-nitrophenyl-?-D-mannopyranoside. PG1711 and PG1712 were ?-1?3 and ?-1?2 mannosidases respectively. No enzyme function could be assigned to PG0973. ?-1?6 mannobiose was not hydrolysed by P. gingivalis W50. EPS present in the culture supernatant was shown to be identical to yeast mannan and a component of the medium used for culturing P. gingivalis and was resistant to hydrolysis by mannosidases. Synthesis of O-LPS and A-LPS and glycosylation of the gingipains appeared to be unaffected in all mutants. Thus ?- and ?-mannosidases of P. gingivalis are not involved in the harnessing of mannan/mannose from the growth medium for these biosynthetic processes. P. gingivalis grown in Chemically Defined Medium devoid of carbohydrate showed reduced ?-mannosidase activity (25%) suggesting these enzymes are environmentally regulated.

Rangarajan M; Aduse-Opoku J; Hashim A; Paramonov N; Curtis MA

2013-09-01

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Blocking Proinflammatory Cytokine Release Modulates Peripheral Blood Mononuclear Cell Response to Porphyromonas gingivalis.  

UK PubMed Central (United Kingdom)

Background: Chronic periodontitis (CP) is an inflammatory disease in which cytokines play a major role in the progression of disease. Anti-inflammatory cytokines (interleukin 4 [IL-4] and IL-10) were reported to be absent or reduced in diseased periodontal tissues, suggesting an imbalance between the proinflammatory and anti-inflammatory mediators. This study tests the hypothesis that there is cellular crosstalk mediated by proinflammatory and anti-inflammatory cytokines and that blocking proinflammatory cytokine (tumor necrosis factor-? [TNF-?] and IL-1) production will enhance anti-inflammatory cytokine (IL-4 and IL-10) production from peripheral blood mononuclear cells (PBMCs) in response to Porphyromonas gingivalis. Methods: PBMCs were isolated from individuals diagnosed with CP or healthy individuals and cultured for 24 hours. Concanavalin A (ConA) was used as an activator of lymphocyte function. Live and heat-killed P. gingivalis or lipopolysaccharide from P. gingivalis were used as the bacterial stimulants. TNF-? and IL-1 production was neutralized by specific antibodies against TNF-? and IL-1? or IL-?. Culture supernatants were evaluated by enzyme-linked immunosorbent assay for TNF-?, IL-1?, IL-4, and IL-10 production. Results: Live P. gingivalis did not result in any significant IL-10 or IL-4 release, whereas heat-killed P. gingivalis led to a significant increase in IL-10 levels compared with unstimulated or live P. gingivalis-stimulated cells from both healthy individuals or those with CP. Overall, PBMCs from patients with CP produced significantly lower IL-10 in response to ConA and P. gingivalis, suggesting chronic suppression of the anti-inflammatory cytokine production. Blocking the proinflammatory cytokine response did not result in any substantial change in IL-10 or IL-4 response to live P. gingivalis. Blocking the proinflammatory cytokine response restored IL-10 production by cells from CP in response to P. gingivalis lipopolysaccharide. Conclusions: These findings suggest that PBMCs from patients with CP have suppressed anti-inflammatory cytokine production that can, in part, be restored by neutralizing proinflammatory cytokines. Monocytes are an important source of IL-10 production, and monocyte-derived IL-10 might play a regulatory role in the pathogenesis of CP.

Berker E; Kantarci A; Hasturk H; Van Dyke TE

2013-09-01

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Blocking Proinflammatory Cytokine Release Modulates Peripheral Blood Mononuclear Cell Response to Porphyromonas gingivalis.  

Science.gov (United States)

Background: Chronic periodontitis (CP) is an inflammatory disease in which cytokines play a major role in the progression of disease. Anti-inflammatory cytokines (interleukin 4 [IL-4] and IL-10) were reported to be absent or reduced in diseased periodontal tissues, suggesting an imbalance between the proinflammatory and anti-inflammatory mediators. This study tests the hypothesis that there is cellular crosstalk mediated by proinflammatory and anti-inflammatory cytokines and that blocking proinflammatory cytokine (tumor necrosis factor-? [TNF-?] and IL-1) production will enhance anti-inflammatory cytokine (IL-4 and IL-10) production from peripheral blood mononuclear cells (PBMCs) in response to Porphyromonas gingivalis. Methods: PBMCs were isolated from individuals diagnosed with CP or healthy individuals and cultured for 24 hours. Concanavalin A (ConA) was used as an activator of lymphocyte function. Live and heat-killed P. gingivalis or lipopolysaccharide from P. gingivalis were used as the bacterial stimulants. TNF-? and IL-1 production was neutralized by specific antibodies against TNF-? and IL-1? or IL-?. Culture supernatants were evaluated by enzyme-linked immunosorbent assay for TNF-?, IL-1?, IL-4, and IL-10 production. Results: Live P. gingivalis did not result in any significant IL-10 or IL-4 release, whereas heat-killed P. gingivalis led to a significant increase in IL-10 levels compared with unstimulated or live P. gingivalis-stimulated cells from both healthy individuals or those with CP. Overall, PBMCs from patients with CP produced significantly lower IL-10 in response to ConA and P. gingivalis, suggesting chronic suppression of the anti-inflammatory cytokine production. Blocking the proinflammatory cytokine response did not result in any substantial change in IL-10 or IL-4 response to live P. gingivalis. Blocking the proinflammatory cytokine response restored IL-10 production by cells from CP in response to P. gingivalis lipopolysaccharide. Conclusions: These findings suggest that PBMCs from patients with CP have suppressed anti-inflammatory cytokine production that can, in part, be restored by neutralizing proinflammatory cytokines. Monocytes are an important source of IL-10 production, and monocyte-derived IL-10 might play a regulatory role in the pathogenesis of CP. PMID:23173823

Berker, Ezel; Kantarci, Alpdogan; Hasturk, Hatice; Van Dyke, Thomas E

2012-11-23

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Prevalence of Porphyromonas gingivalis Four rag Locus Genotypes in Patients of Orthodontic Gingivitis and Periodontitis  

Science.gov (United States)

Porphyromonas gingivalis is considered as a major etiological agent in periodontal diseases and implied to result in gingival inflammation under orthodontic appliance. rag locus is a pathogenicity island found in Porphyromonas gingivalis. Four rag locus variants are different in pathogenicity of Porphyromonas gingivalis. Moreover, there are different racial and geographic differences in distribution of rag locus genotypes. In this study, we assessed the prevalence of Porphyromonas gingivalis and rag locus genotypes in 102 gingival crevicular fluid samples from 57 cases of gingivitis patients with orthodontic appliances, 25 cases of periodontitis patients and 20 cases of periodontally healthy people through a 16S rRNA-based PCR and a multiplex PCR. The correlations between Porphyromona.gingivalis/rag locus and clinical indices were analyzed. The prevalence of Porphyromonas gingivalis and rag locus genes in periodontitis group was the highest among three groups and higher in orthodontic gingivitis than healthy people (pperiodontitis in Shandong. Porphyromonas.gingivalis carrying rag-1 has the strong virulence and could be associated with severe periodontitis.

Liu, Yi; Zhang, Yujie; Wang, Lili; Guo, Yang; Xiao, Shuiqing

2013-01-01

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Benzamidine derivatives inhibit the virulence of Porphyromonas gingivalis.  

Science.gov (United States)

We have previously shown that benzamidine-type compounds can inhibit the activity of arginine-specific cysteine proteinases (gingipains HRgpA and RgpB); well-known virulence factors of Porphyromonas gingivalis. They also hinder in vitro growth of this important periodontopathogenic bacterium. Apparently growth arrest is not associated with their ability to inhibit these proteases, because pentamidine, which is a 20-fold less efficient inhibitor of gingipain than 2,6-bis-(4-amidinobenzyl)-cyclohexanone (ACH), blocked P. gingivalis growth far more effectively. To identify targets for benzamidine-derived compounds other than Arg-gingipains, and to explain their bacteriostatic effects, P. gingivalis ATCC 33277 and P. gingivalis M5-1-2 (clinical isolate) cell extracts were subjected to affinity chromatography using a benzamidine-Sepharose column to identify proteins interacting with benzamidine. In addition to HRgpA and RgpB the analysis revealed heat-shock protein GroEL as another ligand for benzamidine. To better understand the effect of benzamidine-derived compounds on P. gingivalis, bacteria were exposed to benzamidine, pentamidine, ACH and heat, and the expression of gingipains and GroEL was determined. Exposure to heat and benzamidine-derived compounds caused significant increases in GroEL, at both the mRNA and protein levels. Interestingly, despite the fact that gingipains were shown to be the main virulence factors in a fertilized egg model of infection, mortality rates were strongly reduced, not only by ACH, but also by pentamidine, a relatively weak gingipain inhibitor. This effect may depend not only on gingipain inhibition but also on interaction of benzamidine derivatives with GroEL. Therefore these compounds may find use in supportive periodontitis treatment. PMID:23279840

Fröhlich, E; Kantyka, T; Plaza, K; Schmidt, K-H; Pfister, W; Potempa, J; Eick, S

2012-12-26

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Benzamidine derivatives inhibit the virulence of Porphyromonas gingivalis.  

UK PubMed Central (United Kingdom)

We have previously shown that benzamidine-type compounds can inhibit the activity of arginine-specific cysteine proteinases (gingipains HRgpA and RgpB); well-known virulence factors of Porphyromonas gingivalis. They also hinder in vitro growth of this important periodontopathogenic bacterium. Apparently growth arrest is not associated with their ability to inhibit these proteases, because pentamidine, which is a 20-fold less efficient inhibitor of gingipain than 2,6-bis-(4-amidinobenzyl)-cyclohexanone (ACH), blocked P. gingivalis growth far more effectively. To identify targets for benzamidine-derived compounds other than Arg-gingipains, and to explain their bacteriostatic effects, P. gingivalis ATCC 33277 and P. gingivalis M5-1-2 (clinical isolate) cell extracts were subjected to affinity chromatography using a benzamidine-Sepharose column to identify proteins interacting with benzamidine. In addition to HRgpA and RgpB the analysis revealed heat-shock protein GroEL as another ligand for benzamidine. To better understand the effect of benzamidine-derived compounds on P. gingivalis, bacteria were exposed to benzamidine, pentamidine, ACH and heat, and the expression of gingipains and GroEL was determined. Exposure to heat and benzamidine-derived compounds caused significant increases in GroEL, at both the mRNA and protein levels. Interestingly, despite the fact that gingipains were shown to be the main virulence factors in a fertilized egg model of infection, mortality rates were strongly reduced, not only by ACH, but also by pentamidine, a relatively weak gingipain inhibitor. This effect may depend not only on gingipain inhibition but also on interaction of benzamidine derivatives with GroEL. Therefore these compounds may find use in supportive periodontitis treatment.

Fröhlich E; Kantyka T; Plaza K; Schmidt KH; Pfister W; Potempa J; Eick S

2013-06-01

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Involvement of the TREM-1/DAP12 pathway in the innate immune responses to Porphyromonas gingivalis  

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Porphyromonas gingivalis, is a Gram-negative obligate oral anaerobic bacterium highly implicated in periodontal disease, the most prevalent chronic inflammatory disease, but recent evidence also indicates a potential contribution to systemic inflammation. The Triggering Receptor Expressed on Myeloid...

Bostanci, N; Thurnheer, T; Belibasakis, G N

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Immunomagnetic PCR and DNA probe for detection and identification of Porphyromonas gingivalis.  

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The aim of the study that we describe was to combine an immunomagnetic separation and a PCR followed by dot blot hybridization with a DNA probe for the detection and identification of Porphyromonas gingivalis. Immunomagnetic particles were coated with monoclonal antibody specific for P. gingivalis a...

Benkirane, R M; Guillot, E; Mouton, C

39

Immunomagnetic PCR and DNA probe for detection and identification of Porphyromonas gingivalis.  

UK PubMed Central (United Kingdom)

The aim of the study that we describe was to combine an immunomagnetic separation and a PCR followed by dot blot hybridization with a DNA probe for the detection and identification of Porphyromonas gingivalis. Immunomagnetic particles were coated with monoclonal antibody specific for P. gingivalis and were incubated with a suspension containing seven oral bacterial species spiked with various dilutions of P. gingivalis. Beads with their load of bound bacterial were boiled in water, and the target DNA in the supernatant was amplified with a primer pair to generate a 593-bp PCR fragment specific for P. gingivalis. Finally, the product of amplification was detected by dot blot hybridization with a digoxigenin-labeled 593-bp probe. The detection limit was determined to be 100 bacterial cells per ml. The immunomagnetic-PCR/DNA probe procedure described here should be useful for the rapid, specific, and sensitive detection and identification of P. gingivalis in clinical samples.

Benkirane RM; Guillot E; Mouton C

1995-11-01

40

Genetic diversity in the oral pathogen Porphyromonas gingivalis: molecular mechanisms and biological consequences.  

UK PubMed Central (United Kingdom)

Porphyromonas gingivalis is a Gram-negative anaerobic bacterium that colonizes the human oral cavity. It is implicated in the development of periodontitis, a chronic periodontal disease affecting half of the adult population in the USA. To survive in the oral cavity, these bacteria must colonize dental plaque biofilms in competition with other bacterial species. Long-term survival requires P. gingivalis to evade host immune responses, while simultaneously adapting to the changing physiology of the host and to alterations in the plaque biofilm. In reflection of this highly variable niche, P. gingivalis is a genetically diverse species and in this review the authors summarize genetic diversity as it relates to pathogenicity in P. gingivalis. Recent studies revealing a variety of mechanisms by which adaptive changes in genetic content can occur are also reviewed. Understanding the genetic plasticity of P. gingivalis will provide a better framework for understanding the host-microbe interactions associated with periodontal disease.

Tribble GD; Kerr JE; Wang BY

2013-05-01

 
 
 
 
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Characterization of Binding of Streptococcus oralis Glyceraldehyde-3-Phosphate Dehydrogenase to Porphyromonas gingivalis Major Fimbriae  

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Binding of Streptococcus oralis glyceraldehyde-3-phosphate dehydrogenase (GAPDH) to Porphyromonas gingivalis fimbriae was characterized via a biomolecular interaction analysis system. The interaction was specific, and the association constant value was 4.34 × 107 M?1, suggesting that S. oralis GAPDH...

Maeda, Kazuhiko; Nagata, Hideki; Kuboniwa, Masae; Kataoka, Kosuke; Nishida, Nobuko; Tanaka, Muneo; Shizukuishi, Satoshi

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Humoral Responses to Porphyromonas gingivalis Gingipain Adhesin Domains in Subjects with Chronic Periodontitis  

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The gingipains have been implicated in the pathogenicity of Porphyromonas gingivalis, a major etiologic agent of chronic periodontitis. Mature gingipains often present as a membrane-bound glycosylated proteinase-adhesin complex comprising multiple adhesin domains (HA1 to -4) and a catalytic domain. ...

Nguyen, Ky-Anh; DeCarlo, Arthur A.; Paramaesvaran, Mayuri; Collyer, Charles A.; Langley, David B.; Hunter, Neil

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Inhibitory effects of human salivary histatins and lysozyme on coaggregation between Porphyromonas gingivalis and Streptococcus mitis.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The effects of histatins on coaggregation between Porphyromonas gingivalis 381 and Streptococcus mitis ATCC 9811 were investigated by using a turbidimetric assay. The coaggregation activity was significantly inhibited by histatins 5 and 8 and strongly by lysozyme. Tritium-labeled histatin 8 bound to...

Murakami, Y; Nagata, H; Amano, A; Takagaki, M; Shizukuishi, S; Tsunemitsu, A; Aimoto, S

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Detection and comparison of specific hemin binding by Porphyromonas gingivalis and Prevotella intermedia.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

A radioligand assay was designed to detect and compare specific hemin binding by the periodontal anaerobic black-pigmenting bacteria (BPB) Porphyromonas gingivalis and Prevotella intermedia. The assay included physiological concentrations of the hemin-binding protein rabbit serum albumin (RSA) to pr...

Tompkins, G R; Wood, D P; Birchmeier, K R

45

E-selectin mediates Porphyromonas gingivalis adherence to human endothelial cells.  

Science.gov (United States)

Porphyromonas gingivalis, a major periodontal pathogen, may contribute to atherogenesis and other inflammatory cardiovascular diseases. However, little is known about interactions between P. gingivalis and endothelial cells. E-selectin is a membrane protein on endothelial cells that initiates recruitment of leukocytes to inflamed tissue, and it may also play a role in pathogen attachment. In the present study, we examined the role of E-selectin in P. gingivalis adherence to endothelial cells. Human umbilical vein endothelial cells (HUVECs) were stimulated with tumor necrosis factor alpha (TNF-?) to induce E-selectin expression. Adherence of P. gingivalis to HUVECs was measured by fluorescence microscopy. TNF-? increased adherence of wild-type P. gingivalis to HUVECs. Antibodies to E-selectin and sialyl Lewis X suppressed P. gingivalis adherence to stimulated HUVECs. P. gingivalis mutants lacking OmpA-like proteins Pgm6 and -7 had reduced adherence to stimulated HUVECs, but fimbria-deficient mutants were not affected. E-selectin-mediated P. gingivalis adherence activated endothelial exocytosis. These results suggest that the interaction between host E-selectin and pathogen Pgm6/7 mediates P. gingivalis adherence to endothelial cells and may trigger vascular inflammation. PMID:22508864

Komatsu, Toshinori; Nagano, Keiji; Sugiura, Shinsuke; Hagiwara, Makoto; Tanigawa, Naomi; Abiko, Yuki; Yoshimura, Fuminobu; Furuichi, Yasushi; Matsushita, Kenji

2012-04-16

46

E-selectin mediates Porphyromonas gingivalis adherence to human endothelial cells.  

UK PubMed Central (United Kingdom)

Porphyromonas gingivalis, a major periodontal pathogen, may contribute to atherogenesis and other inflammatory cardiovascular diseases. However, little is known about interactions between P. gingivalis and endothelial cells. E-selectin is a membrane protein on endothelial cells that initiates recruitment of leukocytes to inflamed tissue, and it may also play a role in pathogen attachment. In the present study, we examined the role of E-selectin in P. gingivalis adherence to endothelial cells. Human umbilical vein endothelial cells (HUVECs) were stimulated with tumor necrosis factor alpha (TNF-?) to induce E-selectin expression. Adherence of P. gingivalis to HUVECs was measured by fluorescence microscopy. TNF-? increased adherence of wild-type P. gingivalis to HUVECs. Antibodies to E-selectin and sialyl Lewis X suppressed P. gingivalis adherence to stimulated HUVECs. P. gingivalis mutants lacking OmpA-like proteins Pgm6 and -7 had reduced adherence to stimulated HUVECs, but fimbria-deficient mutants were not affected. E-selectin-mediated P. gingivalis adherence activated endothelial exocytosis. These results suggest that the interaction between host E-selectin and pathogen Pgm6/7 mediates P. gingivalis adherence to endothelial cells and may trigger vascular inflammation.

Komatsu T; Nagano K; Sugiura S; Hagiwara M; Tanigawa N; Abiko Y; Yoshimura F; Furuichi Y; Matsushita K

2012-07-01

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Loop-Mediated Isothermal Amplification Method for Rapid Detection of the Periodontopathic Bacteria Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Loop-mediated isothermal amplification (LAMP), a novel nucleic acid amplification method, was developed for the rapid detection of the major periodontal pathogens Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola. The LAMP method amplifies DNA with high specificity, efficiency,...

Yoshida, Akihiro; Nagashima, Shiori; Ansai, Toshihiro; Tachibana, Masayo; Kato, Hiroaki; Watari, Hajime; Notomi, Tsugunori

48

Initial serum antibody titer to Porphyromonas gingivalis influences development of antibody avidity and success of therapy for chronic periodontitis.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

This study assessed the effect of periodontal therapy on specific serum antibody concentration, expressed as titer, and antibody binding strength, expressed as relative avidity. The immune responses to Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans were investigated. Antibody tite...

Mooney, J; Adonogianaki, E; Riggio, M P; Takahashi, K; Haerian, A; Kinane, D F

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Chronic oral infection with Porphyromonas gingivalis accelerates atheroma formation by shifting the lipid profile.  

UK PubMed Central (United Kingdom)

BACKGROUND: Recent studies have suggested that periodontal disease increases the risk of atherothrombotic disease. Atherosclerosis has been characterized as a chronic inflammatory response to cholesterol deposition in the arteries. Although several studies have suggested that certain periodontopathic bacteria accelerate atherogenesis in apolipoprotein E-deficient mice, the mechanistic link between cholesterol accumulation and periodontal infection-induced inflammation is largely unknown. METHODOLOGY/PRINCIPAL FINDINGS: We orally infected C57BL/6 and C57BL/6.KOR-Apoe(shl) (B6.Apoeshl) mice with Porphyromonas gingivalis, which is a representative periodontopathic bacterium, and evaluated atherogenesis, gene expression in the aorta and liver and systemic inflammatory and lipid profiles in the blood. Furthermore, the effect of lipopolysaccharide (LPS) from P. gingivalis on cholesterol transport and the related gene expression was examined in peritoneal macrophages. Alveolar bone resorption and elevation of systemic inflammatory responses were induced in both strains. Despite early changes in the expression of key genes involved in cholesterol turnover, such as liver X receptor and ATP-binding cassette A1, serum lipid profiles did not change with short-term infection. Long-term infection was associated with a reduction in serum high-density lipoprotein (HDL) cholesterol but not with the development of atherosclerotic lesions in wild-type mice. In B6.Apoeshl mice, long-term infection resulted in the elevation of very low-density lipoprotein (VLDL), LDL and total cholesterols in addition to the reduction of HDL cholesterol. This shift in the lipid profile was concomitant with a significant increase in atherosclerotic lesions. Stimulation with P. gingivalis LPS induced the change of cholesterol transport via targeting the expression of LDL receptor-related genes and resulted in the disturbance of regulatory mechanisms of the cholesterol level in macrophages. CONCLUSIONS/SIGNIFICANCE: Periodontal infection itself does not cause atherosclerosis, but it accelerates it by inducing systemic inflammation and deteriorating lipid metabolism, particularly when underlying hyperlidemia or susceptibility to hyperlipidemia exists, and it may contribute to the development of coronary heart disease.

Maekawa T; Takahashi N; Tabeta K; Aoki Y; Miyashita H; Miyauchi S; Miyazawa H; Nakajima T; Yamazaki K

2011-01-01

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Moderate effect of enamel matrix derivative (Emdogain Gel) on Porphyromonas gingivalis growth in vitro.  

UK PubMed Central (United Kingdom)

OBJECTIVE: The aim of this study was to analyse the antibacterial effects of Emdogain Gel or its constituents on the growth of the suspected periodontopathogen Porphyromonas gingivalis. STUDY DESIGN: The effects of the proteins of enamel matrix derivative (EMD), the commercial product Emdogain Gel or its vehicle propylene glycol alginate (PGA) (Straumann, Switzerland) on P. gingivalis growth were determined by two methods: broth dilution assay (BDA) and agar diffusion assay (ADA). RESULTS: BDA-Emdogain Gel inhibited moderately the growth of P. gingivalis, whereas EMD showed no effect. The PGA vehicle inhibited the growth completely. ADA-Emdogain Gel resulted in some inhibition in growth but was not significantly different from control. EMD revealed no zone of inhibition. PGA demonstrated statistically significant zones of inhibition. CONCLUSION: Emdogain Gel shows moderate antibacterial activities against P. gingivalis. These properties seem to be due to the PGA component of the gel preparation.

Walter C; Jawor P; Bernimoulin JP; Hägewald S

2006-03-01

51

Comparison of Real-Time PCR and Culture for Detection of Porphyromonas gingivalis in Subgingival Plaque Samples  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Porphyromonas gingivalis is a major pathogen in destructive periodontal disease in humans. Detection and quantification of this microorganism are relevant for diagnosis and treatment planning. The prevalence and quantity of P. gingivalis in subgingival plaque samples of periodontitis patients were d...

Boutaga, Khalil; van Winkelhoff, Arie Jan; Vandenbroucke-Grauls, Christina M. J. E.; Savelkoul, Paul H. M.

52

The effects of Porphyromonas gingivalis on the cell cycle progression of human gingival epithelial cells.  

UK PubMed Central (United Kingdom)

OBJECTIVE: Porphyromonas gingivalis is a major pathogen in the development and progression of periodontal disease. The interactions or cross-talk between bacteria and gingival epithelial cells drive bacteria to manipulate the cell cycle to favor bacterial survival and virulence expression within the host. This study aims to dissect the effects of P. gingivalis on the cell cycle in human gingival epithelial cells. MATERIALS AND METHODS: We established a model of P. gingivalis invading IHGE cells. The cell cycle distribution of human gingival epithelial cells was analyzed by flow cytometry. Cyclin D and cyclin E mRNA and protein were detected by real-time PCR and Western blot, respectively. RESULTS: Porphyromonas gingivalis-induced facilitation of cell growth was correlated with the acceleration of G1 phase of cell cycle. Cyclin D1 mRNA levels were significantly upregulated from 6 to 12 h after infection. Cyclin E protein and mRNA levels were elevated at 10 and 12 h after invasion. CONCLUSIONS: We confirmed that P. gingivalis significantly enhances IHGE cell proliferation by promoting the G1/S transition, involving the up-regulation of cyclin D and cyclin E.

Pan C; Xu X; Tan L; Lin L; Pan Y

2013-02-01

53

Green tea epigallocatechin-3-gallate attenuates Porphyromonas gingivalis-induced atherosclerosis.  

UK PubMed Central (United Kingdom)

The purpose of this study was to determine whether epigallocatechin-3-gallate (EGCG) ameliorates Porphyromonas gingivalis-induced atherosclerosis. EGCG is a polyphenol extract from green tea with health benefits and P. gingivalis is shown here to accelerate atheroma formation in a murine model. Apolipoprotein E knockout mice were administered EGCG or vehicle in drinking water; they were then fed high-fat diets and injected with P. gingivalis three times a week for 3 weeks. Mice were then killed at 15 weeks. Atherosclerotic plaques in the proximal aorta were determined by Oil Red O staining. Atherosclerosis risk factors in serum, liver or aorta were analysed using cytokine antibody arrays, enzyme-linked immunosorbent assay and real-time PCR. Atherosclerotic lesion areas of the aortic sinus caused by P. gingivalis infection decreased in EGCG-treated groups, wherein EGCG reduced the production of C-reactive protein, monocyte chemoattractant protein-1, and oxidized low-density lipoprotein (LDL), and slightly lowered LDL/very LDL cholesterol in P. gingivalis-challenged mice serum. Furthermore, the increase in CCL2, MMP-9, ICAM-1, HSP60, CD44, LOX-1, NOX-4, p22phox and iNOS gene expression levels in the aorta of P. gingivalis-challenged mice were reduced in EGCG-treated mice. However, HO-1 mRNA levels were elevated by EGCG treatment, suggesting that EGCG, as a natural substance, inhibits P. gingivalis-induced atherosclerosis through anti-inflammatory and antioxidative effects.

Cai Y; Kurita-Ochiai T; Hashizume T; Yamamoto M

2013-02-01

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Influence of titanium on in vitro fibroblast-Porphyromonas gingivalis interaction in peri-implantitis.  

UK PubMed Central (United Kingdom)

AIM: Titanium wear particles have been found in peri-implant tissues, but their role in the pathogenesis of peri-implantitis remains unclear. We aimed to determine the in vitro inflammatory responses of peri-implant granulation tissue fibroblasts (PIGFs) to titanium particles alone and in the presence of viable Porphyromonas gingivalis. MATERIALS & METHODS: Peri-implant granulation tissue fibroblasts were challenged either with TiO2 particles, P. gingivalis or a combination of TiO2 particles and P. gingivalis. Gene expression and protein production of pro-inflammatory mediators by PIGFs were measured with PCR and ELISA, respectively. RESULTS: Higher doses of TiO2 were toxic to PIGFs and in sub-toxic doses, TiO2 caused an increase in gene expression of tumour necrosis factor (TNF)-A and increased protein production of TNF-?, interleukin (IL)-6 and IL-8. A challenge with P. gingivalis alone induced gene expression of TNF-A, IL-1?, IL-6 and IL-8. A combined challenge with TiO2 and P. gingivalis caused a stronger increase in gene expression of TNF-A and protein production of TNF-? and MCP-1 than P. gingivalis alone. CONCLUSIONS: TiO2 particles and P. gingivalis, individually, can induce pro-inflammatory responses in PIGFs. Furthermore, TiO2 particles and viable P. gingivalis further enhance gene expression and production of TNF-? by PIGFs. Therefore, Ti wear particles in the peri-implant tissues in combination with P. gingivalis infection may contribute to the pathogenesis of peri-implantitis by enhancing the inflammation in peri-implant tissues.

Irshad M; Scheres N; Crielaard W; Loos BG; Wismeijer D; Laine ML

2013-09-01

55

[Comparison of periodontal indices and Porphyromonas gingivalis between conventional and self-ligating brackets].  

UK PubMed Central (United Kingdom)

OBJECTIVE: To compare the periodontal indices and Porphyromonas gingivalis (P. gingivalis) between the use of self-ligating brackets and conventional brackets. METHODS: Thirty patients were divided into 2 groups(n=15). Self-ligating brackets were used in the experimental group. Conventional brackets were used in the control group. Clinical periodontal indices, including plaque index (PLI), gingival index (GI) and probing depth (PD) of observed teeth were examined at three different time points: Before orthodontic treatment, the first month after treatment and the third month after treatment. Subgingival plaques were collected simultaneously at each time point. The number of total bacteria and P. gingivalis in each sample were detectd and quantitated by real-time quantitative polymerase chain reaction, the percentage of P. gingivalis in total bacteria was obtained. RESULTS: Before treatment, the periodontal indices and the percentage of P. gingivalis in total bacteria had no difference between the two groups (P>0.05). After 1 and 3 months respectively, the periodontal indices and the percentage of P. gingivalis in total bacteria increased with time (P<0.05) and were obviously lower than those of the control group (P<0.05). CONCLUSION: Compared with conventional brackets, the self-ligating brackets are better for periodontal health. But it is adverse effect on oral health.

Shi J; Liu Y; Hou J; Yan Z; Peng H; Chang X

2013-06-01

56

Deep sequencing of Porphyromonas gingivalis and comparative transcriptome analysis of a LuxS mutant.  

UK PubMed Central (United Kingdom)

Porphyromonas gingivalis is a major etiological agent in chronic and aggressive forms of periodontal disease. The organism is an asaccharolytic anaerobe and is a constituent of mixed species biofilms in a variety of microenvironments in the oral cavity. P. gingivalis expresses a range of virulence factors over which it exerts tight control. High-throughput sequencing technologies provide the opportunity to relate functional genomics to basic biology. In this study we report qualitative and quantitative RNA-Seq analysis of the transcriptome of P. gingivalis. We have also applied RNA-Seq to the transcriptome of a ?luxS mutant of P. gingivalis deficient in AI-2-mediated bacterial communication. The transcriptome analysis confirmed the expression of all predicted ORFs for strain ATCC 33277, including 854 hypothetical proteins, and allowed the identification of hitherto unknown transcriptional units. Twelve non-coding RNAs were identified, including 11 small RNAs and one cobalamin riboswitch. Fifty-seven genes were differentially regulated in the LuxS mutant. Addition of exogenous synthetic 4,5-dihydroxy-2,3-pentanedione (DPD, AI-2 precursor) to the ?luxS mutant culture complemented expression of a subset of genes, indicating that LuxS is involved in both AI-2 signaling and non-signaling dependent systems in P. gingivalis. This work provides an important dataset for future study of P. gingivalis pathophysiology and further defines the LuxS regulon in this oral pathogen.

Hirano T; Beck DA; Demuth DR; Hackett M; Lamont RJ

2012-01-01

57

Deep sequencing of Porphyromonas gingivalis and comparative transcriptome analysis of a LuxS mutant.  

Science.gov (United States)

Porphyromonas gingivalis is a major etiological agent in chronic and aggressive forms of periodontal disease. The organism is an asaccharolytic anaerobe and is a constituent of mixed species biofilms in a variety of microenvironments in the oral cavity. P. gingivalis expresses a range of virulence factors over which it exerts tight control. High-throughput sequencing technologies provide the opportunity to relate functional genomics to basic biology. In this study we report qualitative and quantitative RNA-Seq analysis of the transcriptome of P. gingivalis. We have also applied RNA-Seq to the transcriptome of a ?luxS mutant of P. gingivalis deficient in AI-2-mediated bacterial communication. The transcriptome analysis confirmed the expression of all predicted ORFs for strain ATCC 33277, including 854 hypothetical proteins, and allowed the identification of hitherto unknown transcriptional units. Twelve non-coding RNAs were identified, including 11 small RNAs and one cobalamin riboswitch. Fifty-seven genes were differentially regulated in the LuxS mutant. Addition of exogenous synthetic 4,5-dihydroxy-2,3-pentanedione (DPD, AI-2 precursor) to the ?luxS mutant culture complemented expression of a subset of genes, indicating that LuxS is involved in both AI-2 signaling and non-signaling dependent systems in P. gingivalis. This work provides an important dataset for future study of P. gingivalis pathophysiology and further defines the LuxS regulon in this oral pathogen. PMID:22919670

Hirano, Takanori; Beck, David A C; Demuth, Donald R; Hackett, Murray; Lamont, Richard J

2012-06-12

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Prevalence of Porphyromonas gingivalis and Bacteroides forsythus in Chronic Periodontitis by Multiplex PCR  

Directory of Open Access Journals (Sweden)

Full Text Available The present research decided to study prevalence of Porphyromonas gingivalis and Bacteroides forsythus in chronic periodontitis patient by use of Multiplex PCR. The subgingival plaque samples from 61 patients suffering from chronic periodontitis with probing depth PD?6 and 40 healthy controls were collected by sterile curette. In this study we used two species-specific Forward primers in combination with a single Reverse primer. These primers target variable and conserved region of 16S rRNA gene, respectively. The study included 61 patients (34 women, 27 men; 24-69 years of age; mean 43) and 40 periodontally healthy controls (22 Women, 18 men, 21-69 years in age; mean 41.35%). Porphyromonas gingivalis was detected in 51 samples (83.61%) and 16 samples (40%) of chronic periodontitis patients and healthy subjects, respectively and Bacteroides forsythus was detected in 32 samples (52.50%) of chronic periodontitis patients and was not detected in any sample from healthy persons. We set up Multiplex PCR in order to detect P. gingivalis and B. forsythus simultaneously. The present data suggest that P. gingivalis is a more important cofactor in etiology of chronic periodontitis. Further studies are needed to determine spectrum of pathogenicity of the disease and effective management of diagnosis and treatment in order to decrease the risk of periodontic complicates such as systemic infection.

J. Faghri; Sh. Moghim; A. Moghareh Abed; F. Rezaei; M. Chalabi

2007-01-01

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Polymersome-mediated intracellular delivery of antibiotics to treat Porphyromonas gingivalis-infected oral epithelial cells.  

UK PubMed Central (United Kingdom)

The gram-negative anaerobe Porphyromonas gingivalis colonizes the gingival crevice and is etiologically associated with periodontal disease that can lead to alveolar bone damage and resorption, promoting tooth loss. Although susceptible to antibiotics, P. gingivalis can evade antibiotic killing by residing within gingival keratinocytes. This provides a reservoir of organisms that may recolonize the gingival crevice once antibiotic therapy is complete. Polymersomes are nanosized amphiphilic block copolymer vesicles that can encapsulate drugs. Cells internalize polymersomes by endocytosis into early endosomes, where they are disassembled by the low pH, causing intracellular release of their drug load. In this study, polymersomes were used as vehicles to deliver antibiotics in an attempt to kill intracellular P. gingivalis within monolayers of keratinocytes and organotypic oral mucosal models. Polymersome-encapsulated metronidazole or doxycycline, free metronidazole, or doxycycline, or polymersomes alone as controls, were used, and the number of surviving intracellular P. gingivalis was quantified after host cell lysis. Polymersome-encapsulated metronidazole or doxycycline significantly (P<0.05) reduced the number of intracellular P. gingivalis in both monolayer and organotypic cultures compared to free antibiotic or polymersome alone controls. Polymersomes are effective delivery vehicles for antibiotics that do not normally gain entry to host cells. This approach could be used to treat recurrent periodontitis or other diseases caused by intracellular-dwelling organisms.-Wayakanon, K., Thornhill, M. H., Douglas, C. W. I., Lewis, A. L., Warren, N. J., Pinnock, A., Armes, S. P., Battaglia, G., Murdoch, C. Polymersome-mediated intracellular delivery of antibiotics to treat Porphyromonas gingivalis-infected oral epithelial cells.

Wayakanon K; Thornhill MH; Douglas CW; Lewis AL; Warren NJ; Pinnock A; Armes SP; Battaglia G; Murdoch C

2013-08-01

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Neutrophils alter epithelial response to Porphyromonas gingivalis in a gingival crevice model.  

UK PubMed Central (United Kingdom)

A gingival crevice model (epithelial cell-Porphyromonas gingivalis-neutrophil) was established and used to profile gingipain, matrix metalloproteinase (MMP), MMP mediators [neutrophil gelatinase-associated lipocalin (NGAL) and tissue inhibitor of metalloproteinases 1 (TIMP-1)] and cytokine networks. Smoking is the primary environmental risk factor for periodontitis. Therefore, the influence of cigarette smoke extract (CSE) was also monitored in the same model. Porphyromonas gingivalis alone induced low levels of interleukin-1? and interleukin-8 from epithelial cells, but high levels of both cytokines were produced on the addition of neutrophils. Exposure to CSE (100 and 1000 ng ml(-1) nicotine equivalency) significantly compromised P. gingivalis-induced cytokine secretion (both P < 0.05). P. gingivalis induced impressive secretion of NGAL (P < 0.05) that was not influenced by CSE. The influence of CSE on gingipain production was strain-specific. Purified gingipains effectively and rapidly degraded both TIMP-1 and MMP-9. Induction of large amounts of NGAL, degradation of TIMP-1, and increased gingipain activity would each be expected to prolong collagen degradation and promote disease progression. However, gingipains also degrade MMP-9. Hence, P. gingivalis exerts a complex influence on the proteolytic balance of a gingival crevice model. Exposure to CSE reduces the proinflammatory cytokine burden, which may be expected to promote P. gingivalis survival. In addition to novel findings that provide mechanistic insight into periodontal disease progression, these results are in keeping with the recognized clinical dogma of decreased inflammation/increased disease in smokers. This straightforward gingival crevice model is established as a suitable vehicle for the elucidation of mechanisms that contribute to susceptibility to periodontitis.

Bondy-Carey JL; Galicia J; Bagaitkar J; Potempa JS; Potempa B; Kinane DF; Veillard F; Scott DA

2013-04-01

 
 
 
 
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Porphyromonas gingivalis Regulates TREM-1 in Human Polymorphonuclear Neutrophils via Its Gingipains  

Science.gov (United States)

The Triggering Receptor Expressed on Myeloid cells 1 (TREM-1) is a cell surface receptor of the immunoglobulin superfamily, with the capacity to amplify pro-inflammatory cytokine production and regulate apoptosis. Polymorphonuclear neutrophils (PMNs) are the first line of defence against infection, and a major source of TREM-1. Porphyromonas gingivalis is a Gram-negative anaerobe highly implicated in the inflammatory processes governing periodontal disease, which is characterized by the destruction of the tooth-supporting tissues. It expresses a number of virulence factors, including the cysteine proteinases (or gingipains). The aim of this in vitro study was to investigate the effect of P. gingivalis on TREM-1 expression and production by primary human PMNs, and to evaluate the role of its gingipains in this process. After 4 h of challenge, P. gingivalis enhanced TREM-1 expression as identified by quantitative real-time PCR. This was followed by an increase in soluble (s)TREM-1 secretion over a period of 18 h, as determined by ELISA. At this time-point, the P. gingivalis-challenged PMNs exhibited diminished TREM-1 cell-membrane staining, as identified by flow cytometry and confocal laser scanning microscopy. Furthermore engagement of TREM-1, by means of anti-TREM-1 antibodies, enhanced the capacity of P. gingivalis to stimulate interleukin (IL)-8 production. Conversely, antagonism of TREM-1 using a synthetic peptide resulted in reduction of IL-8 secretion. Using isogenic P. gingivalis mutant strains, we identified the Arg-gingipain to be responsible for shedding of sTREM-1 from the PMN surface, whereas the Lys-gingipain had the capacity to degrade TREM-1. In conclusion, the differential regulation of TREM-1 by the P. gingivalis gingipains may present a novel mechanism by which P. gingivalis manipulates the host innate immune response helping to establish chronic periodontal inflammation.

Bostanci, Nagihan; Thurnheer, Thomas; Aduse-Opoku, Joseph; Curtis, Michael A.; Zinkernagel, Annelies S.; Belibasakis, Georgios N.

2013-01-01

62

Effects of nicotine on the growth and protein expression of Porphyromonas gingivalis.  

UK PubMed Central (United Kingdom)

Tobacco smoking is considered one of the most significant environmental risk factors for destructive periodontal disease. The effect of smoking on periodontopathic microbiota has not yet been elucidated, as previous studies failed to identify a concrete relationship between periodontopathic microorganisms and smoking. However, it is likely that smoking, as an environmental stress factor, may affect the behavior of dental plaque microorganisms, ultimately leading to alteration of the host-parasite interaction. The goal of this study was to examine the effect of nicotine, a major component of tobacco, on the growth and protein expression of the crucial periodontal pathogen Porphyromonas gingivalis. The growth of P. gingivalis 381 was measured after bacterial cells were cultivated in liquid broth containing various nicotine concentrations. First, P. gingivalis cells were allowed to grow in the presence of a single dose of nicotine (the single exposure protocol) at 0, 1, 2, 4, and 8 mg/L, respectively. Second, P. gingivalis cells were exposed to five consecutive doses of nicotine (the multiple exposure protocol) at 0, 1, 2, and 4 mg/L, respectively. Bacterial growth was measured by optical density and protein expression was analyzed by SDS-PAGE and 2-D gel electrophoresis. In the single nicotine exposure protocol, it was observed that the growth of P. gingivalis 381 was inhibited by nicotine in a dose-dependent manner. In the multiple nicotine exposure protocol, the growth rate of P. gingivalis increased with each subsequent nicotine exposure, even though bacterial growth was also inhibited in a dose dependent fashion. SDS-PAGE and 2-D gel electrophoresis analyses revealed a minor change in the pattern of protein expression, showing differences in proteins with low molecular weights (around 20 kDa) on exposure to nicotine. The results of this study suggest that nicotine exerts an inhibitory effect on the growth of P. gingivalis, and has a potential to modulate protein expression in P. gingivalis.

Baek O; Zhu W; Kim HC; Lee SW

2012-02-01

63

HtrA in Porphyromonas gingivalis can regulate growth and gingipain activity under stressful environmental conditions.  

UK PubMed Central (United Kingdom)

In several micro-organisms, HtrA, a serine periplasmic protease, is considered an important virulence factor that plays a regulatory role in oxidative and temperature stress. The authors have previously shown that the vimA gene product is an important virulence regulator in Porphyromonas gingivalis. Further, purified recombinant VimA physically interacted with the major gingipains and the HtrA from P. gingivalis. To further evaluate a role for HtrA in the pathogenicity of this organism, a 1.5 kb fragment containing the htrA gene was PCR-amplified from the chromosomal DNA of P. gingivalis W83. This gene was insertionally inactivated using the ermF-ermAM antibiotic-resistance cassette and used to create an htrA-deficient mutant by allelic exchange. In one randomly chosen isogenic mutant designated P. gingivalis FLL203, there was increased sensitivity to hydrogen peroxide. Growth of this mutant at an elevated temperature was more inhibited compared to the wild-type. Further, in contrast to the wild-type, there was a significant decrease in Arg-gingipain activity after heat shock in FLL203. However, the gingipain activity in the mutant returned to normal levels after a further 30 min incubation at room temperature. Collectively, these data suggest that HtrA may play a similar role in oxidative and temperature stress in P. gingivalis as observed in other organisms.

Roy F; Vanterpool E; Fletcher HM

2006-11-01

64

The atherogenic bacterium Porphyromonas gingivalis evades circulating phagocytes by adhering to erythrocytes  

DEFF Research Database (Denmark)

A relationship between periodontitis and coronary heart disease has been investigated intensively. A pathogenic role for the oral bacterium Porphyromonas gingivalis has been suggested for both diseases. We examined whether complement activation by P. gingivalis strain ATCC 33277 allows the bacterium to adhere to human red blood cells (RBCs) and thereby evade attack by circulating phagocytes. On incubation with normal human serum, the P. gingivalis strain efficiently fixed complement component 3 (C3). Incubation of bacteria with washed whole blood cells suspended in autologous serum resulted in a dose- and time-dependent adherence to RBCs. The adherence required functionally intact complement receptor 1 (CR1; also called CD35) on the RBCs and significantly inhibited the uptake of P. gingivalis by neutrophils and B cells within 1 min of incubation (by 64% and 51%, respectively) and that by monocytes after between 15 min and 30 min of incubation (by 66% and 53%, respectively). The attachment of C3b/iC3b to bacterium-bearing RBCs decreased progressively after 15 min, indicating that conversion of C3 fragments into C3dg occurred, decreasing the affinity for CR1 on RBCs. We propose that P. gingivalis exploits RBCs as a transport vehicle, rendering it inaccessible to attack by phagocytes, and by doing so plays a role in the development of systemic diseases.

BelstrØm, Daniel; Holmstrup, Palle

2011-01-01

65

Identification and characterization of novel glycoproteins involved in growth and biofilm formation by Porphyromonas gingivalis.  

Science.gov (United States)

Porphyromonas gingivalis has been implicated as a major pathogen associated with chronic periodontitis. To extend our knowledge of post-translational protein glycosylation in P. gingivalis, a proteomic analysis involving two-dimensional polyacrylamide gel electrophoresis combined with carbohydrate staining and mass spectrometry was performed. Four novel glycoproteins, PGN0743, PGN0876, PGN1513 and PGN0729, in P. gingivalis ATCC 33277 were identified. These four identified glycoproteins possess a range of biochemical activities and cellular localization. PGN0743 contains a sequence motif identifying it as a FKBP-type cis-trans isomerase, which has activity usually associated with chaperone functions. PGN0876 and PGN1513 contain tetratricopeptide repeat domains that mediate protein-protein interactions. PGN0729 encodes the outer membrane protein 41 precursor, which was previously identified as Pgm6, and is homologous to the OmpA protein in Escherichia coli. Several different types of glycoprotein were identified, suggesting that P. gingivalis possesses a general mechanism for protein glycosylation. PGN0743-deficient and PGN0876-deficient mutants were constructed to examine the role(s) of the two identified glycoproteins. Both mutants showed a decreased growth rate under nutrient-limited conditions and reduced biofilm formation activity. These results suggest that the novel glycoproteins PGN0743 and PGN0876 play an important role in the growth and colonization of P. gingivalis. PMID:23134611

Kishi, M; Hasegawa, Y; Nagano, K; Nakamura, H; Murakami, Y; Yoshimura, F

2012-07-21

66

Identification and characterization of novel glycoproteins involved in growth and biofilm formation by Porphyromonas gingivalis.  

UK PubMed Central (United Kingdom)

Porphyromonas gingivalis has been implicated as a major pathogen associated with chronic periodontitis. To extend our knowledge of post-translational protein glycosylation in P. gingivalis, a proteomic analysis involving two-dimensional polyacrylamide gel electrophoresis combined with carbohydrate staining and mass spectrometry was performed. Four novel glycoproteins, PGN0743, PGN0876, PGN1513 and PGN0729, in P. gingivalis ATCC 33277 were identified. These four identified glycoproteins possess a range of biochemical activities and cellular localization. PGN0743 contains a sequence motif identifying it as a FKBP-type cis-trans isomerase, which has activity usually associated with chaperone functions. PGN0876 and PGN1513 contain tetratricopeptide repeat domains that mediate protein-protein interactions. PGN0729 encodes the outer membrane protein 41 precursor, which was previously identified as Pgm6, and is homologous to the OmpA protein in Escherichia coli. Several different types of glycoprotein were identified, suggesting that P. gingivalis possesses a general mechanism for protein glycosylation. PGN0743-deficient and PGN0876-deficient mutants were constructed to examine the role(s) of the two identified glycoproteins. Both mutants showed a decreased growth rate under nutrient-limited conditions and reduced biofilm formation activity. These results suggest that the novel glycoproteins PGN0743 and PGN0876 play an important role in the growth and colonization of P. gingivalis.

Kishi M; Hasegawa Y; Nagano K; Nakamura H; Murakami Y; Yoshimura F

2012-12-01

67

Modelación por homología de la proteína Luxs de Porphyromonas gingivalis cepa W83/ Modelling by homology of Luxs protein in Porphyromonas gingivalis strain W83  

Scientific Electronic Library Online (English)

Full Text Available Abstract in spanish Antecedentes: En las proteínas no se logra siempre su cristalización, de buen tamaño y de buena calidad para someterla a difracción de rayos X. De tal manera que se abre un campo para el desarrollo de estudios teóricos moleculares y proteínicos, que permiten la representación de las moléculas en tres dimensiones, proporcionando una información espacial para estudiar la interacción entre ligandos y receptores macromoleculares. Materialesy Métodos: Estudio In sil (more) ico, a partir del análisis de secuencias primarias de seis diferentes proteínas LuxS cristalizadas de diversas bacterias, se seleccionó la proteína 1J6X del Helicobacter pylori, por su similaridad con la secuencia de la proteína LuxS en Porphyromonas gingivalis (P. gingivalis) cepa W83, para producir un modelo por homología de esta proteína, utilizando los programas Sybyl y MOE. Se realizó un acoplamiento con el ligando natural para evaluar la reproducibilidad del modelo en un ambiente biológico. Resultados: Se desarrolló el modelado de la proteína LuxS de P. gingivalis cepa W83, que permite el acercamiento a una estructura que se propone, por la interacción entre la proteína y su ligando natural. El modelo generado con recursos computacionales logró una correcta estructura molecular que aceptó la realización de diversos cálculos. El acoplamiento demostró una cavidad donde se logran diversas posiciones del ligando con buenos resultados. Conclusiones: Se obtuvo un modelo 3D para la proteína LuxS en la P. gingivalis cepa W83 validado por diferentes métodos computacionales con una adecuada reproducibilidad biológica por medio del acoplamiento molecular. Abstract in english Background: Crystallization is not always achieved for all proteins in a good size and a good quality for X-ray diffraction. So that condition opens a field for the development of theoretical molecular and protein studies allowing the representation of the molecules in 3D, providing spatial information to study the interaction between ligands and macromolecular receptors. Materials and Methods: In silico study from primary sequence analysis of six different proteins LuxS (more) crystallized of several bacteria. 1J6X protein of Helicobacter pylori was selected for its similarity with the LuxS protein sequence in Porphyromonas gingivalis (P. gingivalis) strain W83 to produce a homology model of this protein, using the Sybyl and MOE software. A docking was performed to assess the reproducibility of the model in a biological environment. Results: The LuxS protein modelling of P. gingivalis strain W83 was developed, which allows the approach to a proposed structure for the interaction between the protein and its natural ligand. The model generated with computational resources achieved the correct position and biological behavior by means of developed calculations. The docking showed a cavity in which the ligand adopted several positions with good results. Conclusions: A LuxS protein model was obtained, validated by different methods. This generated a 3D model for LuxS protein in P. gingivalis strain W83 with biological reproducibility by means of molecular docking.

Díaz Caballero, A; Martínez Serrano, E; Vivas Reyes, R; Puerta Llerena, L; Méndez Cuadro, D; Cabrales Salgado, R; Padilla Rodríguez, A

2012-12-01

68

Modelación por homología de la proteína Luxs de Porphyromonas gingivalis cepa W83 Modelling by homology of Luxs protein in Porphyromonas gingivalis strain W83  

Directory of Open Access Journals (Sweden)

Full Text Available Antecedentes: En las proteínas no se logra siempre su cristalización, de buen tamaño y de buena calidad para someterla a difracción de rayos X. De tal manera que se abre un campo para el desarrollo de estudios teóricos moleculares y proteínicos, que permiten la representación de las moléculas en tres dimensiones, proporcionando una información espacial para estudiar la interacción entre ligandos y receptores macromoleculares. Materialesy Métodos: Estudio In silico, a partir del análisis de secuencias primarias de seis diferentes proteínas LuxS cristalizadas de diversas bacterias, se seleccionó la proteína 1J6X del Helicobacter pylori, por su similaridad con la secuencia de la proteína LuxS en Porphyromonas gingivalis (P. gingivalis) cepa W83, para producir un modelo por homología de esta proteína, utilizando los programas Sybyl y MOE. Se realizó un acoplamiento con el ligando natural para evaluar la reproducibilidad del modelo en un ambiente biológico. Resultados: Se desarrolló el modelado de la proteína LuxS de P. gingivalis cepa W83, que permite el acercamiento a una estructura que se propone, por la interacción entre la proteína y su ligando natural. El modelo generado con recursos computacionales logró una correcta estructura molecular que aceptó la realización de diversos cálculos. El acoplamiento demostró una cavidad donde se logran diversas posiciones del ligando con buenos resultados. Conclusiones: Se obtuvo un modelo 3D para la proteína LuxS en la P. gingivalis cepa W83 validado por diferentes métodos computacionales con una adecuada reproducibilidad biológica por medio del acoplamiento molecular.Background: Crystallization is not always achieved for all proteins in a good size and a good quality for X-ray diffraction. So that condition opens a field for the development of theoretical molecular and protein studies allowing the representation of the molecules in 3D, providing spatial information to study the interaction between ligands and macromolecular receptors. Materials and Methods: In silico study from primary sequence analysis of six different proteins LuxS crystallized of several bacteria. 1J6X protein of Helicobacter pylori was selected for its similarity with the LuxS protein sequence in Porphyromonas gingivalis (P. gingivalis) strain W83 to produce a homology model of this protein, using the Sybyl and MOE software. A docking was performed to assess the reproducibility of the model in a biological environment. Results: The LuxS protein modelling of P. gingivalis strain W83 was developed, which allows the approach to a proposed structure for the interaction between the protein and its natural ligand. The model generated with computational resources achieved the correct position and biological behavior by means of developed calculations. The docking showed a cavity in which the ligand adopted several positions with good results. Conclusions: A LuxS protein model was obtained, validated by different methods. This generated a 3D model for LuxS protein in P. gingivalis strain W83 with biological reproducibility by means of molecular docking.

A Díaz Caballero; E Martínez Serrano; R Vivas Reyes; L Puerta Llerena; D Méndez Cuadro; R Cabrales Salgado; A Padilla Rodríguez

2012-01-01

69

The unique hmuY gene sequence as a specific marker of Porphyromonas gingivalis.  

UK PubMed Central (United Kingdom)

Porphyromonas gingivalis, a major etiological agent of chronic periodontitis, acquires heme from host hemoproteins using the HmuY hemophore. The aim of this study was to develop a specific P. gingivalis marker based on a hmuY gene sequence. Subgingival samples were collected from 66 patients with chronic periodontitis and 40 healthy subjects and the entire hmuY gene was analyzed in positive samples. Phylogenetic analyses demonstrated that both the amino acid sequence of the HmuY protein and the nucleotide sequence of the hmuY gene are unique among P. gingivalis strains/isolates and show low identity to sequences found in other species (below 50 and 56%, respectively). In agreement with these findings, a set of hmuY gene-based primers and standard/real-time PCR with SYBR Green chemistry allowed us to specifically detect P. gingivalis in patients with chronic periodontitis (77.3%) and healthy subjects (20%), the latter possessing lower number of P. gingivalis cells and total bacterial cells. Isolates from healthy subjects possess the hmuY gene-based nucleotide sequence pattern occurring in W83/W50/A7436 (n?=?4), 381/ATCC 33277 (n?=?3) or TDC60 (n?=?1) strains, whereas those from patients typically have TDC60 (n?=?21), W83/W50/A7436 (n?=?17) and 381/ATCC 33277 (n?=?13) strains. We observed a significant correlation between periodontal index of risk of infectiousness (PIRI) and the presence/absence of P. gingivalis (regardless of the hmuY gene-based sequence pattern of the isolate identified [r?=?0.43; P?=?0.0002] and considering particular isolate pattern [r?=?0.38; P?=?0.0012]). In conclusion, we demonstrated that the hmuY gene sequence or its fragments may be used as one of the molecular markers of P. gingivalis.

Gmiterek A; Wójtowicz H; Mackiewicz P; Radwan-Oczko M; Kantorowicz M; Chomyszyn-Gajewska M; Fr?szczak M; Bielecki M; Olczak M; Olczak T

2013-01-01

70

The unique hmuY gene sequence as a specific marker of Porphyromonas gingivalis.  

Science.gov (United States)

Porphyromonas gingivalis, a major etiological agent of chronic periodontitis, acquires heme from host hemoproteins using the HmuY hemophore. The aim of this study was to develop a specific P. gingivalis marker based on a hmuY gene sequence. Subgingival samples were collected from 66 patients with chronic periodontitis and 40 healthy subjects and the entire hmuY gene was analyzed in positive samples. Phylogenetic analyses demonstrated that both the amino acid sequence of the HmuY protein and the nucleotide sequence of the hmuY gene are unique among P. gingivalis strains/isolates and show low identity to sequences found in other species (below 50 and 56%, respectively). In agreement with these findings, a set of hmuY gene-based primers and standard/real-time PCR with SYBR Green chemistry allowed us to specifically detect P. gingivalis in patients with chronic periodontitis (77.3%) and healthy subjects (20%), the latter possessing lower number of P. gingivalis cells and total bacterial cells. Isolates from healthy subjects possess the hmuY gene-based nucleotide sequence pattern occurring in W83/W50/A7436 (n?=?4), 381/ATCC 33277 (n?=?3) or TDC60 (n?=?1) strains, whereas those from patients typically have TDC60 (n?=?21), W83/W50/A7436 (n?=?17) and 381/ATCC 33277 (n?=?13) strains. We observed a significant correlation between periodontal index of risk of infectiousness (PIRI) and the presence/absence of P. gingivalis (regardless of the hmuY gene-based sequence pattern of the isolate identified [r?=?0.43; P?=?0.0002] and considering particular isolate pattern [r?=?0.38; P?=?0.0012]). In conclusion, we demonstrated that the hmuY gene sequence or its fragments may be used as one of the molecular markers of P. gingivalis. PMID:23844074

Gmiterek, Anna; Wójtowicz, Halina; Mackiewicz, Pawe?; Radwan-Oczko, Ma?gorzata; Kantorowicz, Ma?gorzata; Chomyszyn-Gajewska, Maria; Fr?szczak, Magdalena; Bielecki, Marcin; Olczak, Mariusz; Olczak, Teresa

2013-07-02

71

The Unique hmuY Gene Sequence as a Specific Marker of Porphyromonas gingivalis  

Science.gov (United States)

Porphyromonas gingivalis, a major etiological agent of chronic periodontitis, acquires heme from host hemoproteins using the HmuY hemophore. The aim of this study was to develop a specific P. gingivalis marker based on a hmuY gene sequence. Subgingival samples were collected from 66 patients with chronic periodontitis and 40 healthy subjects and the entire hmuY gene was analyzed in positive samples. Phylogenetic analyses demonstrated that both the amino acid sequence of the HmuY protein and the nucleotide sequence of the hmuY gene are unique among P. gingivalis strains/isolates and show low identity to sequences found in other species (below 50 and 56%, respectively). In agreement with these findings, a set of hmuY gene-based primers and standard/real-time PCR with SYBR Green chemistry allowed us to specifically detect P. gingivalis in patients with chronic periodontitis (77.3%) and healthy subjects (20%), the latter possessing lower number of P. gingivalis cells and total bacterial cells. Isolates from healthy subjects possess the hmuY gene-based nucleotide sequence pattern occurring in W83/W50/A7436 (n?=?4), 381/ATCC 33277 (n?=?3) or TDC60 (n?=?1) strains, whereas those from patients typically have TDC60 (n?=?21), W83/W50/A7436 (n?=?17) and 381/ATCC 33277 (n?=?13) strains. We observed a significant correlation between periodontal index of risk of infectiousness (PIRI) and the presence/absence of P. gingivalis (regardless of the hmuY gene-based sequence pattern of the isolate identified [r?=?0.43; P?=?0.0002] and considering particular isolate pattern [r?=?0.38; P?=?0.0012]). In conclusion, we demonstrated that the hmuY gene sequence or its fragments may be used as one of the molecular markers of P. gingivalis.

Mackiewicz, Pawel; Radwan-Oczko, Malgorzata; Kantorowicz, Malgorzata; Chomyszyn-Gajewska, Maria; Fraszczak, Magdalena; Bielecki, Marcin; Olczak, Mariusz; Olczak, Teresa

2013-01-01

72

Intracellular survival and vascular cell-to-cell transmission of Porphyromonas gingivalis  

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Full Text Available Abstract Background Porphyromonas gingivalis is associated with periodontal disease and invades different cell types including epithelial, endothelial and smooth muscle cells. In addition to P. gingivalis DNA, we have previously identified live invasive bacteria in atheromatous tissue. However, the mechanism of persistence of this organism in vascular tissues remains unclear. Therefore, the objective of this study was to analyze the ability of intracellular P. gingivalis to persist for extended periods of time, transmit to and possibly replicate in different cell types. Results Using antibiotic protection assays, immunofluorescent and laser confocal microscopy, we found that after a prolonged intracellular phase, while P. gingivalis can still be detected by immunostaining, the intracellular organisms lose their ability to be recovered in vitro. Surprisingly however, intracellular P. gingivalis could be recovered in vitro upon co incubation with fresh vascular host cells. We then demonstrated that the organism was able to exit the initially infected host cells, then enter and multiply in new host cells. Further, we found that cell-to-cell contact increased the transmission rate but was not required for transmission. Finally, we found that the invasion of new host cells allowed P. gingivalis to increase its numbers. Conclusion Our results suggest that the persistence of vascular tissue-embedded P. gingivalis is due to its ability to transmit among different cell types. This is the first communication demonstrating the intercellular transmission as a likely mechanism converting latent intracellular bacteria from state of dormancy to a viable state allowing for persistence of an inflammatory pathogen in vascular tissue.

Li Ling; Michel Raynald; Cohen Joshua; DeCarlo Arthur; Kozarov Emil

2008-01-01

73

Functional differences of Porphyromonas gingivalis Fimbriae in determining periodontal disease pathogenesis: a literature review/ Prevalencia de los genotipos FimA de Porphyromonas gingivalis en diferentes poblaciones mundiales: Revisión de la Literatura  

Scientific Electronic Library Online (English)

Full Text Available Abstract in spanish Porphyromonas gingivalis es un microorganismo implicado en la periodontitis crónica y agresiva. Dentro de sus factores de virulencia, se encuentran las Fimbrias, las cuales están compuestas por una proteína denominada FimBrillina, que está codificada por el gen FimA, del cual existen 6 genotipos (I, II, III, IV, V, Ib), según la secuencia de nucleótidos. Los genotipos II y IV han sido relacionados con periodontitis, mientras el I con salud periodontal. Identificar l (more) os genotipos de FimA de P. gingivalis en pacientes con periodontitis podría generar nuevas estrategias que conlleven a suprimir los genotipos más patogénicos para prevenir el desarrollo de la periodontitis en portadores sanos. Se revisó la prevalencia de los genotipos de FimA de P. gingivalis en diferentes países del mundo, para lo cual se realizó una búsqueda sistemática en bases de datos de Pubmed, Hinary y Science Direct usando los descriptores: Porphyromonas gingivalis, adhesión bacteriana, periodontitis, Fimbrias, Fim A, y genotipificación hasta abril del 2011. Abstract in english Porphyromonas gingivalis is implicated in chronic and aggressive periodontitis. This bacterium has numerous virulence factors and one is the Fimbriae, which is quite important for bacterial colonization. Fimbriae are appendices that anchor to the bacterial wall and are comprised of the protein FimBriline encoded by the FimA gene. Thus far, six genotypes have been identified, FimA I to V and Ib. Genotypes II and IV are associated with periodontal disease, while genotype I (more) is related to gingival health. Genotype identification of P. gingivalis FimA in periodontitis would be important to confirm the pathogenic genotypes and to establish risk at population level. This review is about the P. gingivalis FimA genotype prevalence worldwide. A systematic search using Pubmed, Hinary, and Science Direct within the following descriptors: Porphyromonas gingivalis, bacterial adhesion, periodontitis, Fimbriae, FimA, genotipification was performed to April 2011.

Moreno, Sandra; Contreras, Adolfo

2013-01-01

74

Potential value of a rice protein extract, containing proteinaceous inhibitors against cysteine proteinases from Porphyromonas gingivalis, for managing periodontal diseases.  

UK PubMed Central (United Kingdom)

Arg-specific gingipain (Rgp) is a major pathogenic determinant of Porphyromonas gingivalis which is a major pathogen in periodontal disease. We prepared protein extracts with Rgp-inhibitory activity from polished rice (Oryza sativa) and evaluated the effects of these extracts on the growth and pathogenicity of P. gingivalis. The extracts inhibited the proteolytic degradation of human proteins by P. gingivalis proteinases, and repressed the growth and homotypic biofilm formation of P. gingivalis. The disruption of adhesion of epithelial cells by P. gingivalis was also restricted by the rice protein extracts. Our results suggested that the rice protein extracts suppressed the pathogenicity and growth of P. gingivalis by inhibiting the bacterial proteinase activities, implying that the Rgp-inhibitory proteins prepared from rice may be potentially valuable as nutraceutical agents for preventing periodontal diseases.

Taiyoji M; Yamanaka T; Tsuno T; Ohtsubo S

2013-01-01

75

Potential value of a rice protein extract, containing proteinaceous inhibitors against cysteine proteinases from Porphyromonas gingivalis, for managing periodontal diseases.  

Science.gov (United States)

Arg-specific gingipain (Rgp) is a major pathogenic determinant of Porphyromonas gingivalis which is a major pathogen in periodontal disease. We prepared protein extracts with Rgp-inhibitory activity from polished rice (Oryza sativa) and evaluated the effects of these extracts on the growth and pathogenicity of P. gingivalis. The extracts inhibited the proteolytic degradation of human proteins by P. gingivalis proteinases, and repressed the growth and homotypic biofilm formation of P. gingivalis. The disruption of adhesion of epithelial cells by P. gingivalis was also restricted by the rice protein extracts. Our results suggested that the rice protein extracts suppressed the pathogenicity and growth of P. gingivalis by inhibiting the bacterial proteinase activities, implying that the Rgp-inhibitory proteins prepared from rice may be potentially valuable as nutraceutical agents for preventing periodontal diseases. PMID:23291749

Taiyoji, Mayumi; Yamanaka, Takashi; Tsuno, Takuo; Ohtsubo, Sadami

2013-01-07

76

Asociación entre porphyromona gingivalis y proteína C reactiva en enfermedades sistémicas inflamatorias Association between porphyromonas gingivalis and C-reactive protein in systemic inflammatory diseases  

Directory of Open Access Journals (Sweden)

Full Text Available La proteína C reactiva (PCR) es un marcador serológico de la inflamación asociado con incremento en el riesgo de enfermedades sistémicas inflamatorias (ESI). La periodontitis también se relaciona con niveles elevados de PCR en adultos y con una reducción de la misma después de su tratamiento. Así, se ha postulado que la PCR puede ser un posible mediador de la asociación entre periodontitis y ESI. Los patógenos periodontales además de inducir inflamación local y destrucción tisular están involucrados en el aumento de la respuesta sistémica inflamatoria e inmunológica. Diferentes autores han investigado la relación entre los anticuerpos para algunos patógenos periodontales y la PCR, pero la asociación se ha notificado firmemente para IgG a Porphyromona gingivalis. Es escasa la evidencia de asociación de una medida directa entre patógenos periodontales y PCR, sin embargo es muy importante debido a que la presencia de anticuerpos no necesariamente es un indicador de infección activa.C-reactive protein (CRP) is a serological marker of systemic inflammation that has been associated with increased risk systemic inflammatory diseases. Periodontitis has also been linked to elevated CRP levels in adults as well as with a reduction in PCR after its treatment. It is thus postulated that CRP might be a possible mediator of the association between periodontitis and systemic inflammatory diseases. Periodontal pathogens do not induce only local inflammation and tissue destruction. They are also involved in systemic increases in inflammatory and inmmune responses. Several studies have investigated antibodies to various periodontal pathogens in relation to CRP, but the association has been reported consistently only for IgG to Porphyromonas gingivalis. Evidence is sparse on the association between a direct measure of periodontal pathogens and CRP, while it is more important because the presence of antibody titers is not necessarily indicative of an active infection.

C.M. Ardila Medina; G.I. Lafaurie Villamil

2010-01-01

77

Humoral immune response to antigens of Porphyromonas gingivalis ATCC 33277 in chronic periodontitis  

Scientific Electronic Library Online (English)

Full Text Available Abstract in english Introduction: Periodontitis is a chronic disease that results from an interaction of a mixed bacterial challenge and the host response. Objective: The purposes of this study were to evaluate the IgG serum levels to Porphyromonas gingivalis antigens by ELISA in individuals with different periodontal conditions correlated with clinical parameters, and to analyze the immunoreactivity profiles by Western blotting. Methods: Serum IgG levels against the cell sonicate antigen fr (more) om 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. Results: In the CP group, sera reactivity by ELISA was significantly higher than in the G and H groups (Kruskal-Wallis p

Franca, Mônica; Moura-Costa, Lília; Meyer, Roberto J.; Trindade, Soraya C.; Tunes, Urbino da Rocha; Freire, Songelí M.

2007-06-01

78

Hyperlipidemia Impaired Innate Immune Response to Periodontal Pathogen Porphyromonas gingivalis in Apolipoprotein E Knockout Mice  

Science.gov (United States)

A finely-tuned innate immune response plays a pivotal role in protecting host against bacterial invasion during periodontal disease progression. Hyperlipidemia has been suggested to exacerbate periodontal health condition. However, the underlying mechanism has not been addressed. In the present study, we investigated the effect of hyperlipidemia on innate immune responses to periodontal pathogen Porphyromonas gingivalis infection. Apolipoprotein E-deficient and wild-type mice at the age of 20 weeks were used for the study. Peritoneal macrophages were isolated and subsequently used for the study of viable P. gingivalis infection. ApoE?/? mice demonstrated inhibited iNOS production and impaired clearance of P. gingivalis in vitro and in vivo; furthermore, ApoE?/? mice displayed disrupted cytokine production pattern in response to P. gingivalis, with a decreased production of tumor necrosis factor-?, interleukin-6 (IL-6), IL-1? and monocyte chemotactic protein-1. Microarray data demonstrated that Toll-like receptor (TLR) and NOD-like receptor (NLR) pathway were altered in ApoE?/? mice macrophages; further analysis of pattern recognition receptors (PRRs) demonstrated that expression of triggering receptors on myeloid cells-1 (TREM-1), an amplifier of the TLR and NLR pathway, was decreased in ApoE?/? mice macrophages, leading to decreased recruitment of NF-?B onto the promoters of the TNF-? and IL-6. Our data suggest that in ApoE?/? mice hyperlipidemia disrupts the expression of PRRs, and cripples the host’s capability to generate sufficient innate immune response to P. gingivalis, which may facilitate immune evasion, subgingival colonization and establishment of P. gingivalis in the periodontal niche.

Yan, Fuhua; Xiao, Yin

2013-01-01

79

Adhesion of Porphyromonas gingivalis and Biofilm Formation on Different Types of Orthodontic Brackets.  

UK PubMed Central (United Kingdom)

Objectives. To examine the interaction between Porphyromonas gingivalis and 3 different orthodontic brackets in vitro, focusing on the effect of an early salivary pellicle and other bacteria on the formation of biofilms. Material and Methods. Mono- and multi-species P. gingivalis biofilms were allowed to form in vitro, on 3 different bracket types (stainless steel, ceramic and plastic) with and without an early salivary pellicle. The brackets were anaerobically incubated for 3 days in Brain Heart Infusion Broth to form biofilms. Bacteria were quantified by trypsin treatment and enumeration of the total viable counts of bacteria recovered. Results. Saliva was found to significantly affect (P < 0.001) adhesion and biofilm formation of P. gingivalis, with higher numbers for the coated brackets. No significant effect was detected for the impact of the type of biofilm, although on stainless steel and plastic brackets there was a tendency for higher numbers of the pathogen in multi-species biofilms. Bracket material alone was not found to affect the number of bacteria. Conclusions. The salivary pellicle seems to facilitate the adhesion of P. gingivalis and biofilm formation on orthodontic brackets, while the material comprising the brackets does not significantly impact on the number of bacteria.

Papaioannou W; Panagopoulos A; Koletsi-Kounari H; Kontou E; Makou M

2012-01-01

80

?-Amylase is a potential growth inhibitor of Porphyromonas gingivalis, a periodontal pathogenic bacterium.  

Science.gov (United States)

BACKGROUND AND OBJECTIVE: Porphyromonas gingivalis is a major etiological agent in the development and progression of periodontal diseases. In this study, we isolated a cell growth inhibitor against P. gingivalis species from rice protein extract. MATERIAL AND METHODS: The cell growth inhibitor active against P. gingivalis was purified from polished rice extract using a six-step column chromatography process. Its antimicrobial properties were investigated through microscope analysis, spectrum of activity and general structure. RESULTS: The inhibitor was identified as AmyI-1, an ?-amylase, and showed significant cell growth inhibitory activity against P. gingivalis species. Scanning electron microscopy micrograph analysis and bactericidal assay indicated an intriguing possibility that the inhibitor compromises the cell membrane structure of the bacterial cells and leads to cell death. Moreover, ?-amylases from human saliva and porcine pancreas showed inhibitory activity similar to that of AmyI-1. CONCLUSIONS: This is the first study to report that ?-amylases cause cell death of periodontal pathogenic bacteria. This finding highlights the potential importance and therapeutic potential of ?-amylases in treating periodontal diseases. PMID:23550921

Ochiai, A; Harada, K; Hashimoto, K; Shibata, K; Ishiyama, Y; Mitsui, T; Tanaka, T; Taniguchi, M

2013-04-01

 
 
 
 
81

?-Amylase is a potential growth inhibitor of Porphyromonas gingivalis, a periodontal pathogenic bacterium.  

UK PubMed Central (United Kingdom)

BACKGROUND AND OBJECTIVE: Porphyromonas gingivalis is a major etiological agent in the development and progression of periodontal diseases. In this study, we isolated a cell growth inhibitor against P. gingivalis species from rice protein extract. MATERIAL AND METHODS: The cell growth inhibitor active against P. gingivalis was purified from polished rice extract using a six-step column chromatography process. Its antimicrobial properties were investigated through microscope analysis, spectrum of activity and general structure. RESULTS: The inhibitor was identified as AmyI-1, an ?-amylase, and showed significant cell growth inhibitory activity against P. gingivalis species. Scanning electron microscopy micrograph analysis and bactericidal assay indicated an intriguing possibility that the inhibitor compromises the cell membrane structure of the bacterial cells and leads to cell death. Moreover, ?-amylases from human saliva and porcine pancreas showed inhibitory activity similar to that of AmyI-1. CONCLUSIONS: This is the first study to report that ?-amylases cause cell death of periodontal pathogenic bacteria. This finding highlights the potential importance and therapeutic potential of ?-amylases in treating periodontal diseases.

Ochiai A; Harada K; Hashimoto K; Shibata K; Ishiyama Y; Mitsui T; Tanaka T; Taniguchi M

2013-04-01

82

Prevalence of fimA genotypes of Porphyromonas gingivalis and other periodontal bacteria in a Spanish population with chronic periodontitis  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Objectives: The aim of this study was to determine the prevalence of the different fimA genotypes of Porphyromonas gingivalis in adult Spanish patients with chronic periodontitis, patients with gingivitis and periodontally healthy subjects, and the relationship between these genotypes and other peri...

Puig-Silla, Miriam; Dasí-Fernánde, Francisco; Montiel-Company, José-María; Almerich-Silla, José-Manuel

83

Genomic comparison of invasive and rare non-invasive strains reveals Porphyromonas gingivalis genetic polymorphisms  

Directory of Open Access Journals (Sweden)

Full Text Available Porphyromonas gingivalis strains are shown to invade human cells in vitro with different invasion efficiencies, varying by up to three orders of magnitude.We tested the hypothesis that invasion-associated interstrain genomic polymorphisms are present in P. gingivalis and that putative invasion-associated genes can contribute to P. gingivalis invasion.Using an invasive (W83) and the only available non-invasive P. gingivalis strain (AJW4) and whole genome microarrays followed by two separate software tools, we carried out comparative genomic hybridization (CGH) analysis.We identified 68 annotated and 51 hypothetical open reading frames (ORFs) that are polymorphic between these strains. Among these are surface proteins, lipoproteins, capsular polysaccharide biosynthesis enzymes, regulatory and immunoreactive proteins, integrases, and transposases often with abnormal GC content and clustered on the chromosome. Amplification of selected ORFs was used to validate the approach and the selection. Eleven clinical strains were investigated for the presence of selected ORFs. The putative invasion-associated ORFs were present in 10 of the isolates. The invasion ability of three isogenic mutants, carrying deletions in PG0185, PG0186, and PG0982 was tested. The PG0185 (ragA) and PG0186 (ragB) mutants had 5.1×103-fold and 3.6×103-fold decreased in vitro invasion ability, respectively.The annotation of divergent ORFs suggests deficiency in multiple genes as a basis for P. gingivalis non-invasive phenotype. Access the supplementary material to this article: Supplement, table (see Supplementary files under Reading Tools online).

Svetlana Dolgilevich; Brian Rafferty; Darya Luchinskaya; Emil Kozarov

2011-01-01

84

Role of the Streptococcus gordonii SspB protein in the development of Porphyromonas gingivalis biofilms on streptococcal substrates.  

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Porphyromonas gingivalis is an aggressive periodontal pathogen that persists in the mixed-species plaque biofilm on tooth surfaces. P. gingivalis cells attach to the plaque commensal Streptococcus gordonii and this coadhesion event leads to the development of P. gingivalis biofilms. Binding of these organisms is multimodal, involving both the P. gingivalis major fimbrial FimA protein and the species-specific interaction of the minor fimbrial Mfa1 protein with the streptococcal SspB protein. This study examined the contribution of the Mfa1-SspB interaction to P. gingivalis biofilm formation. P. gingivalis biofilms readily formed on substrata of S. gordonii DL1 but not on Streptococcus mutans cells which lack a coadhesion-mediating homologue of SspB. An insertional inactivation of the mfa1 gene in P. gingivalis resulted in a phenotype deficient in S. gordonii binding and unable to form biofilms. Furthermore, analysis using recombinant streptococci and enterococci showed that P. gingivalis biofilms formed on Enterococcus faecalis strains expressing SspB or translational fusions of SspB with SpaP (the non-adherent SspB homologue in S. mutans) containing the P. gingivalis adherence domain (SspB adherence region, BAR) of SspB. In contrast, an isogenic Ssp null mutant of S. gordonii DL1 was unable to support biofilm growth, even though this strain bound to P. gingivalis FimA at levels similar to wild-type S. gordonii DL1. Finally, site-specific mutation of two functional amino acid residues in BAR resulted in SspB polypeptides that did not promote the development of P. gingivalis biofilms. These results suggest that the induction of P. gingivalis biofilms on a streptococcal substrate requires functional SspB-minor fimbriae interactions. PMID:12055284

Lamont, Richard J; El-Sabaeny, Azza; Park, Yoonsuk; Cook, Guy S; Costerton, J William; Demuth, Donald R

2002-06-01

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regT can modulate gingipain activity and response to oxidative stress in Porphyromonas gingivalis.  

UK PubMed Central (United Kingdom)

Recombinant VimA protein can interact with the gingipains and several other proteins that may play a role in its biogenesis in Porphyromonas gingivalis. In silico analysis of PG2096, a hypothetical protein that was shown to interact with VimA, suggests that it may have environmental stress resistance properties. To further evaluate the role(s) of PG2096, the predicted open reading frame was PCR amplified from P. gingivalis W83 and insertionally inactivated using the ermF-ermAM antibiotic-resistance cassette. One randomly chosen PG2096-defective mutant created by allelic exchange and designated FLL205 was further characterized. Under normal growth conditions at 37 °C, Arg-X and Lys-X gingipain activities in FLL205 were reduced by approximately 35?% and 21?%, respectively, compared to the wild-type strain. However, during prolonged growth at an elevated temperature of 42 °C, Arg-X activity was increased by more than 40?% in FLL205 in comparison to the wild-type strain. In addition, the PG2096-defective mutant was more resistant to oxidative stress when treated with 0.25 mM hydrogen peroxide. Taken together these results suggest that the PG2096 gene, designated regT (regulator of gingipain activity at elevated temperatures), may be involved in regulating gingipain activity at elevated temperatures and be important in oxidative stress resistance in P. gingivalis.

Vanterpool E; Aruni AW; Roy F; Fletcher HM

2010-10-01

86

Antibacterial activity against Porphyromonas gingivalis and biological characteristics of antibacterial stainless steel.  

UK PubMed Central (United Kingdom)

To evaluate the possibility of an alternative to the traditional orthodontic stainless steel implants, the antibacterial activity against Porphyromonas gingivalis (P. gingivalis) and the related cytotoxicity of a type 304 Cu bearing antibacterial stainless steel were studied. The results indicated that the antibacterial stainless steel showed excellent antibacterial property against P. gingivalis, compared with the control steel (a purchased medical grade 304 stainless steel). Compared to the control steel, there were fewer bacteria on the surface of the antibacterial stainless steel, with significant difference in morphology. The cytotoxicities of the antibacterial stainless steel to both MG-63 and KB cells were all grade 1, the same as those of the control steel. There were no significant differences in the apoptosis rates on MG-63 and KB cells between the antibacterial stainless steel and the control steel. This study demonstrates that the antibacterial stainless steel is possible to reduce the incidence of implant-related infections and can be a more suitable material for the micro-implant than the conventional stainless steel in orthodontic treatment.

Zhang D; Ren L; Zhang Y; Xue N; Yang K; Zhong M

2013-05-01

87

Effectiveness of Antibiotic Medicaments against Biofilm Formation of Enterococcus faecalis and Porphyromonas gingivalis.  

UK PubMed Central (United Kingdom)

INTRODUCTION: In this study we compared the antibacterial effect of triple antibiotic paste (TAP), double antibiotic paste (DAP), and calcium hydroxide [Ca(OH)2] against Enterococcus faecalis and Porphyromonas gingivalis biofilm. METHODS: The minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), minimum biofilm inhibitory concentration (MBIC), and biofilm formation were measured by using microtiter plate methods. The 2 bacteria were treated with different dilutions of TAP, DAP, and Ca(OH)2 solutions. The turbidities of the bacterial cultures in the microtiter plate were measured by optical density at 490 nm by using a spectrophotometer. Data were analyzed by 2-way analysis of variance (? = 0.05). RESULTS: For TAP, the MIC and MBIC values were 0.003 mg/mL for E. faecalis and 0.006 mg/mL for P. gingivalis. The MBC values for TAP were 0.3 mg/mL for both bacteria. The MIC and MBIC values for DAP were 0.001 mg/mL for E. faecalis and P. gingivalis. The MBC values for DAP were 0.14 mg/mL for both bacteria. Biofilm formation of the 2 bacteria was significantly decreased with TAP and DAP at all tested dilutions (P < .0001) compared with control groups; however, TAP and DAP biofilm formations were not significantly different from each other. Ca(OH)2 significantly decreased bacterial biofilm formation compared with the control, but it was significantly less than TAP and DAP (P < .05). CONCLUSIONS: Both TAP and DAP were more effective than Ca(OH)2 against E. faecalis and P. gingivalis. DAP can be considered an effective and comparable antibacterial substitute for TAP.

Sabrah AH; Yassen GH; Gregory RL

2013-11-01

88

Erythritol alters microstructure and metabolomic profiles of biofilm composed of Streptococcus gordonii and Porphyromonas gingivalis.  

UK PubMed Central (United Kingdom)

The effects of sugar alcohols such as erythritol, xylitol, and sorbitol on periodontopathic biofilm are poorly understood, though they have often been reported to be non-cariogenic sweeteners. In the present study, we evaluated the efficacy of sugar alcohols for inhibiting periodontopathic biofilm formation using a heterotypic biofilm model composed of an oral inhabitant Streptococcus gordonii and a periodontal pathogen Porphyromonas gingivalis. Confocal microscopic observations showed that the most effective reagent to reduce P. gingivalis accumulation onto an S. gordonii substratum was erythritol, as compared with xylitol and sorbitol. In addition, erythritol moderately suppressed S. gordonii monotypic biofilm formation. To examine the inhibitory effects of erythritol, we analyzed the metabolomic profiles of erythritol-treated P. gingivalis and S. gordonii cells. Metabolome analyses using capillary electrophoresis time-of-flight mass spectrometry revealed that a number of nucleic intermediates and constituents of the extracellular matrix, such as nucleotide sugars, were decreased by erythritol in a dose-dependent manner. Next, comparative analyses of metabolites of erythritol- and sorbitol-treated cells were performed using both organisms to determine the erythritol-specific effects. In P. gingivalis, all detected dipeptides, including Glu-Glu, Ser-Glu, Tyr-Glu, Ala-Ala and Thr-Asp, were significantly decreased by erythritol, whereas they tended to be increased by sorbitol. Meanwhile, sorbitol promoted trehalose 6-phosphate accumulation in S. gordonii cells. These results suggest that erythritol has inhibitory effects on dual species biofilm development via several pathways, including suppression of growth resulting from DNA and RNA depletion, attenuated extracellular matrix production, and alterations of dipeptide acquisition and amino acid metabolism.

Hashino E; Kuboniwa M; Alghamdi SA; Yamaguchi M; Yamamoto R; Cho H; Amano A

2013-07-01

89

The effects of Mg-ion implantation and sandblasting on Porphyromonas gingivalis attachment.  

UK PubMed Central (United Kingdom)

OBJECTIVES: The purpose of this study was to evaluate the effect of titanium surface treatment on Porphyromonas gingivalis bacterial attachment. MATERIALS AND METHODS: Titanium disks of 15 mm in diameter and 1 mm in thickness (n=40) were subjected to mechanical grinding, or sandblasting. Magnesium (Mg) ions were implanted onto the titanium surface using a plasma source ion implantation method. The structure, chemistry, and surface morphologies of the titanium surfaces were analyzed using scanning electron microscopy (SEM), X-ray photoelectron spectroscopy and Auger electron spectroscopy. Surface roughness was measured using a laser profilometer. Half of the titanium disks in each group were dipped in saliva for 24 h. All of the titanium specimens were rinsed with distilled water. A P. gingivalis strain was cultured in anaerobic conditions at 37°C for 72 h, and all titanium specimens were dipped in the bacterial suspension at 37°C for 24 h. Specimens were examined at × 3000 magnification using a SEM. The number of bacteria in each of 10 separate fields was determined by directly counting the number of bacterial colonies that adhered to each specimen. The mean values were calculated afterward. The resulting data were analyzed to assess the significance of observed differences based on the method of the surface treatment, ion implantation, and saliva dipping. RESULTS: The amount of P. gingivalis attached to the sandblasted specimens was greater than that on the ground specimens (P<0.001). Moreover, surfaces with Mg-ion implantation had more attachments than nonimplanted surfaces (P<0.001). Saliva dipping acted synergistically with surface roughness and chemical composition of the specimens. CONCLUSIONS: Chemically modified surface increase the attachment of a major periodontopathic bacterium, P. gingivalis.

Kim ML; Jeong CM; Jeon YC; Byon E; Jeong Y; Cho LR

2012-02-01

90

Capsaicin inhibits Porphyromonas gingivalis growth, biofilm formation, gingivomucosal inflammatory cytokine secretion, and in vitro osteoclastogenesis.  

UK PubMed Central (United Kingdom)

The prevention and treatment of periodontitis requires not only the control of causative pathogens, especially Porphyromonas gingivalis, but also the regulation of inflammatory immune response. Investigating auxiliary drugs for periodontitis during conventional treatments is, thus, quite important. Capsaicin, an agonist for the vanilloid receptor subtype 1 (TRPV1), due to its bacteriostatic activity against Gram-negative bacteria and anti-inflammatory effects, appears to be a promising drug. In this work, the antimicrobial activity of capsaicin against P. gingivalis and biofilm formation, inflammatory cytokine levels in experimental periodontitis, osteoclast precursor proliferation, and osteoclastogenesis in vitro were fully investigated. The results showed that capsaicin inhibited P. gingivalis growth with a minimum inhibitory concentration (MIC) and a minimum bactericidal concentration (MBC) of 16 and 64 mg/l, respectively. Capsaicin also inhibited P. gingivalis biofilm formation, with minimum biofilm inhibition concentrations MBIC50 and MBIC90 of 16 and 32 mg/l, respectively, and reduced pre-formed biofilms' viability with a minimum biofilm reduction concentration MBRC50 of 64 mg/l, as demonstrated by confocal laser scanning microscopy. In experimental periodontitis, except for IL-10, TNF-?, IL-1?, IL-6, IL-12, and iNOS were depressed after capsaicin treatment. Moreover, capsaicin also suppressed osteoclast precursor proliferation and osteoclastogenesis, as demonstrated by NF-?B p65. However, this favorable effect was attenuated by the TRPV1 antagonist, camphor. It, thus, suggests that capsaicin is a potential drug for the auxiliary treatment of periodontitis. TRPV1 activation may involve in beneficial roles of capsaicin on periodontitis.

Zhou Y; Guan X; Zhu W; Liu Z; Wang X; Yu H; Wang H

2013-08-01

91

Humoral immune response to antigens of Porphyromonas gingivalis ATCC 33277 in chronic periodontitis  

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Full Text Available Introduction: Periodontitis is a chronic disease that results from an interaction of a mixed bacterial challenge and the host response. Objective: The purposes of this study were to evaluate the IgG serum levels to Porphyromonas gingivalis antigens by ELISA in individuals with different periodontal conditions correlated with clinical parameters, and to analyze the immunoreactivity profiles by Western blotting. Methods: 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. Results: 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). There was no statistically significant difference between G and HP reactivity (Dunnett t3 p=0.617). Among individuals with chronic periodontitis, the IgG-anti-P. gingivalis serum levels were positively correlated with percentage of clinical attachment level =5mm (r s = + 0.375, p<0.05) and a negative correlation was found between IgG-anti-P. gingivalis levels and percentage of probing pocket depth 0-3mm (r s = - 0. 411, p< 0.05). The analysis of sera immunoreactivity profiles to sonicate antigen by Western blotting showed differences between the sera of CP, G and H group individuals. The serum from CP frequently reacted with high molecular weight (103 kDa, 86 kDa, 72 kDa, 60 kDa, 58 kDa, 52 kDa) protein fractions. Conclusions: Serum levels of IgG anti-P. gingivalis distinguished individuals with chronic periodontitis, gingivitis and healthy periodontium. There was a correlation between clinical parameters and serum IgG levels against P. gingivalis. There was a difference in the recognition profile of protein fractions among the studied groups and some bands were more specific

Mônica Franca; Lília Moura-Costa; Roberto J. Meyer; Soraya C. Trindade; Urbino da Rocha Tunes; Songelí M. Freire

2007-01-01

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

UK PubMed Central (United Kingdom)

BACKGROUND: Apoptosis is a highly controlled process of cell death that can be induced by periodontopathogens. The present study aimed to investigate the expression of Fas and Bcl-2 proteins by CD3+ T cells in vitro under stimulation by total Porphyromonas gingivalis antigens and purified recombinant P. gingivalis HmuY protein. RESULTS: CD3+ T cells derived from CP patients and stimulated with HmuY expressed higher levels of Bcl-2 compared to identical cells stimulated with P. gingivalis crude extract or cells derived from NP control subjects (p = 0.043). CONCLUSION: The authors hypothesize that P. gingivalis HmuY plays a role in the pathogenesis of chronic periodontitis, possibly by reducing or delaying apoptosis in T cells through a pathway involving the Bcl-2 protein.

Carvalho-Filho PC; Trindade SC; Olczak T; Sampaio GP; Oliveira-Neto MG; Santos HA; Pereira BF; Moura-Costa L; Xavier MT; Meyer R

2013-09-01

93

A Two-Component System Regulates Hemin Acquisition in Porphyromonas gingivalis.  

UK PubMed Central (United Kingdom)

Porphyromonas gingivalis is a Gram-negative oral anaerobe associated with infection of the periodontia. The organism has a small number of two-component signal transduction systems, and after comparing genome sequences of strains W83 and ATCC 33277 we discovered that the latter was mutant in histidine kinase (PGN_0752), while the cognate response regulator (PGN_0753) remained intact. Microarray-based transcriptional profiling and ChIP-seq assays were carried out with an ATCC 33277 transconjugant containing the functional histidine kinase from strain W83 (PG0719). The data showed that the regulon of this signal transduction system contained genes that were involved in hemin acquisition, including gingipains, at least three transport systems, as well as being self-regulated. Direct regulation by the response regulator was confirmed by electrophoretic mobility shift assays. In addition, the system appears to be activated by hemin and the regulator acts as both an activator and repressor.

Scott JC; Klein BA; Duran-Pinedo A; Hu L; Duncan MJ

2013-01-01

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A Two-Component System Regulates Hemin Acquisition in Porphyromonas gingivalis.  

Science.gov (United States)

Porphyromonas gingivalis is a Gram-negative oral anaerobe associated with infection of the periodontia. The organism has a small number of two-component signal transduction systems, and after comparing genome sequences of strains W83 and ATCC 33277 we discovered that the latter was mutant in histidine kinase (PGN_0752), while the cognate response regulator (PGN_0753) remained intact. Microarray-based transcriptional profiling and ChIP-seq assays were carried out with an ATCC 33277 transconjugant containing the functional histidine kinase from strain W83 (PG0719). The data showed that the regulon of this signal transduction system contained genes that were involved in hemin acquisition, including gingipains, at least three transport systems, as well as being self-regulated. Direct regulation by the response regulator was confirmed by electrophoretic mobility shift assays. In addition, the system appears to be activated by hemin and the regulator acts as both an activator and repressor. PMID:24039921

Scott, Jodie C; Klein, Brian A; Duran-Pinedo, Ana; Hu, Linden; Duncan, Margaret J

2013-09-05

95

The periodontal pathogen Porphyromonas gingivalis cleaves apoB-100 and increases the expression of apoM in LDL in whole blood leading to cell proliferation  

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Objective: Several studies support an association between periodontal disease and atherosclerosis with a crucial role for the pathogen Porphyromonas gingivalis. This study aims to investigate the proteolytic and oxidative activity of P. gingivalis on LDL in a whole blood system by using a proteomic ...

Bengtsson, Torbjörn; Karlsson, Helen; Gunnarsson, Patrik; Skoglund, Caroline; Elison, Charlotte; Leanderson, Per

96

Maturation of the Arginine-Specific Proteases of Porphyromonas gingivalis W50 Is Dependent on a Functional prR2 Protease Gene  

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The prpR1 of Porphyromonas gingivalis codes for three distinct enzymes with specificity for arginyl peptide bonds termed RI, RIA, and RIB. These three isoforms comprise the majority of the extracellular, arginine-specific protease activity in P. gingivalis W50. RI is a heterodimer in which the catal...

Aduse-Opoku, Joseph; Rangarajan, Minnie; Young, Katherine A.; Curtis, Michael A.

97

vimA gene downstream of recA is involved in virulence modulation in Porphyromonas gingivalis W83.  

Science.gov (United States)

A 0.9-kb open reading frame encoding a unique 32-kDa protein was identified downstream of the recA gene of Porphyromonas gingivalis. Reverse transcription-PCR and Northern blot analysis showed that both the recA gene and this open reading frame are part of the same transcriptional unit. This cloned fragment was insertionally inactivated using the ermF-ermAM antibiotic resistance cassette to create a defective mutant by allelic exchange. When plated on Brucella blood agar, the mutant strain, designated P. gingivalis FLL92, was non-black pigmented and showed significant reduction in beta-hemolysis compared with the parent strain, P. gingivalis W83. Arginine- and lysine-specific cysteine protease activities, which were mostly soluble, were approximately 90% lower than that of the parent strain. Expression of the rgpA, rgpB, and kgp protease genes was the same in P. gingivalis FLL92 as in the wild-type strain. In contrast to the parent strain, P. gingivalis FLL92 showed increased autoaggregration in addition to a significant reduction in hemagglutinating and hemolysin activities. In in vivo experiments using a mouse model, P. gingivalis FLL92 was dramatically less virulent than the parent strain. A molecular survey of this mutant and the parent strain using all known P. gingivalis insertion sequence elements as probes suggested that no intragenomic changes due to the movement of these elements have occurred in P. gingivalis FLL92. Taken together, these results suggest that the recA downstream gene, designated vimA (virulence-modulating gene), plays an important role in virulence modulation in P. gingivalis W83, possibly representing a novel posttranscriptional or translational regulation of virulence factors in P. gingivalis. PMID:11119521

Abaibou, H; Chen, Z; Olango, G J; Liu, Y; Edwards, J; Fletcher, H M

2001-01-01

98

vimA gene downstream of recA is involved in virulence modulation in Porphyromonas gingivalis W83.  

UK PubMed Central (United Kingdom)

A 0.9-kb open reading frame encoding a unique 32-kDa protein was identified downstream of the recA gene of Porphyromonas gingivalis. Reverse transcription-PCR and Northern blot analysis showed that both the recA gene and this open reading frame are part of the same transcriptional unit. This cloned fragment was insertionally inactivated using the ermF-ermAM antibiotic resistance cassette to create a defective mutant by allelic exchange. When plated on Brucella blood agar, the mutant strain, designated P. gingivalis FLL92, was non-black pigmented and showed significant reduction in beta-hemolysis compared with the parent strain, P. gingivalis W83. Arginine- and lysine-specific cysteine protease activities, which were mostly soluble, were approximately 90% lower than that of the parent strain. Expression of the rgpA, rgpB, and kgp protease genes was the same in P. gingivalis FLL92 as in the wild-type strain. In contrast to the parent strain, P. gingivalis FLL92 showed increased autoaggregration in addition to a significant reduction in hemagglutinating and hemolysin activities. In in vivo experiments using a mouse model, P. gingivalis FLL92 was dramatically less virulent than the parent strain. A molecular survey of this mutant and the parent strain using all known P. gingivalis insertion sequence elements as probes suggested that no intragenomic changes due to the movement of these elements have occurred in P. gingivalis FLL92. Taken together, these results suggest that the recA downstream gene, designated vimA (virulence-modulating gene), plays an important role in virulence modulation in P. gingivalis W83, possibly representing a novel posttranscriptional or translational regulation of virulence factors in P. gingivalis.

Abaibou H; Chen Z; Olango GJ; Liu Y; Edwards J; Fletcher HM

2001-01-01

99

Identification and characterization of Porphyromonas gingivalis client proteins that bind to Streptococcus oralis glyceraldehyde-3-phosphate dehydrogenase.  

UK PubMed Central (United Kingdom)

Coaggregation of Porphyromonas gingivalis and oral streptococci is thought to play an important role in P. gingivalis colonization. Previously, we reported that P. gingivalis major fimbriae interacted with Streptococcus oralis glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and that amino acid residues 166 to 183 of GAPDH exhibited strong binding activity toward P. gingivalis fimbriae (H. Nagata, M. Iwasaki, K. Maeda, M. Kuboniwa, E. Hashino, M. Toe, N. Minamino, H. Kuwahara, and S. Shizukuishi, Infect. Immun. 77:5130-5138, 2009). The present study aimed to identify and characterize P. gingivalis components other than fimbriae that interact with S. oralis GAPDH. A pulldown assay was performed to detect potential interactions between P. gingivalis client proteins and S. oralis recombinant GAPDH with amino acid residues 166 to 183 deleted by site-directed mutagenesis. Seven proteins, namely, tonB-dependent receptor protein (RagA4), arginine-specific proteinase B, 4-hydroxybutyryl-coenzyme A dehydratase (AbfD), lysine-specific proteinase, GAPDH, NAD-dependent glutamate dehydrogenase (GDH), and malate dehydrogenase (MDH), were identified by two-dimensional gel electrophoresis followed by proteomic analysis using tandem mass spectrometry. Interactions between these client proteins and S. oralis GAPDH were analyzed with a biomolecular interaction analysis system. S. oralis GAPDH showed high affinity for five of the seven client proteins (RagA4, AbfD, GAPDH, GDH, and MDH). Interactions between P. gingivalis and S. oralis were measured by a turbidimetric method and fluorescence microscopy. RagA4, AbfD, and GDH enhanced coaggregation, whereas GAPDH and MDH inhibited coaggregation. Furthermore, the expression of luxS in P. gingivalis was upregulated by RagA4, AbfD, and GDH but was downregulated by MDH. These results indicate that the five P. gingivalis client proteins function as regulators in P. gingivalis biofilm formation with oral streptococci.

Maeda K; Nagata H; Kuboniwa M; Ojima M; Osaki T; Minamino N; Amano A

2013-03-01

100

Hepatocytes produce tumor necrosis factor-? and interleukin-6 in response to Porphyromonas gingivalis.  

UK PubMed Central (United Kingdom)

BACKGROUND AND OBJECTIVE: The liver plays a major role in clearing systemic bacterial infections. In addition, inflammatory cytokines produced in the liver play a critical role in systemic cytokine levels. The aim of this study was to investigate the production of tumor necrosis factor-? (TNF-?) and interleukin-6 (IL-6) by hepatocytes in response to periodontal pathogens. MATERIAL AND METHODS: The mouse hepatic carcinoma cell line Hepa-1.6 and the mouse macrophage-like cell line RAW 264 were co-cultured in Transwell insert plates. Cells were stimulated with bacterial extracts prepared from Porphyromonas gingivalis and the induction of TNF-? and IL-6 was measured using real-time PCR and ELISA. RESULTS: After stimulation with bacteria, the induction of TNF-? and IL-6 was observed in RAW 264 cells and Hepa-1.6 cells. Significant reduction of TNF-? mRNA expression in Hepa-1.6 cells was observed after treatment with antibody to TNF-?. CONCLUSION: The results obtained in the present study show that P. gingivalis extract induces TNF-? and IL-6 in an in vitro liver model and that macrophage-derived TNF-? mediates the induction of TNF-? in hepatocytes.

Takano M; Sugano N; Mochizuki S; Koshi RN; Narukawa TS; Sawamoto Y; Ito K

2012-02-01

 
 
 
 
101

Expression, purification and characterization of enoyl-ACP reductase II, FabK, from Porphyromonas gingivalis  

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The rapid rise in bacterial drug resistance coupled with the low number of novel antimicrobial compounds in the discovery pipeline has led to a critical situation requiring the expedient discovery and characterization of new antimicrobial drug targets. Enzymes in the bacterial fatty acid synthesis pathway, FAS-II, are distinct from their mammalian counterparts, FAS-I, in terms of both structure and mechanism. As such, they represent attractive targets for the design of novel antimicrobial compounds. Enoyl-acyl carrier protein reductase II, FabK, is a key, rate-limiting enzyme in the FAS-II pathway for several bacterial pathogens. The organism, Porphyromonas gingivalis, is a causative agent of chronic periodontitis that affects up to 25% of the US population and incurs a high national burden in terms of cost of treatment. P. gingivalis expresses FabK as the sole enoyl reductase enzyme in its FAS-II cycle, which makes this a particularly appealing target with potential for selective antimicrobial therapy. Herein we report the molecular cloning, expression, purification and characterization of the FabK enzyme from P. gingivalis, only the second organism from which this enzyme has been isolated. Characterization studies have shown that the enzyme is a flavoprotein, the reaction dependent upon FMN and NADPH and proceeding via a Ping-Pong Bi-Bi mechanism to reduce the enoyl substrate. A sensitive assay measuring the fluorescence decrease of NADPH as it is converted to NADP{sup +} during the reaction has been optimized for high-throughput screening. Finally, protein crystallization conditions have been identified which led to protein crystals that diffract x-rays to high resolution.

Hevener, Kirk E.; Mehboob, Shahila; Boci, Teuta; Truong, Kent; Santarsiero, Bernard D.; Johnson, Michael E. (UIC)

2012-10-25

102

Porphyromonas gingivalis Facilitates the Development and Progression of Destructive Arthritis through Its Unique Bacterial Peptidylarginine Deiminase (PAD)  

Science.gov (United States)

Rheumatoid arthritis and periodontitis are two prevalent chronic inflammatory diseases in humans and are associated with each other both clinically and epidemiologically. Recent findings suggest a causative link between periodontal infection and rheumatoid arthritis via bacteria-dependent induction of a pathogenic autoimmune response to citrullinated epitopes. Here we showed that infection with viable periodontal pathogen Porphyromonas gingivalis strain W83 exacerbated collagen-induced arthritis (CIA) in a mouse model, as manifested by earlier onset, accelerated progression and enhanced severity of the disease, including significantly increased bone and cartilage destruction. The ability of P. gingivalis to augment CIA was dependent on the expression of a unique P. gingivalis peptidylarginine deiminase (PPAD), which converts arginine residues in proteins to citrulline. Infection with wild type P. gingivalis was responsible for significantly increased levels of autoantibodies to collagen type II and citrullinated epitopes as a PPAD-null mutant did not elicit similar host response. High level of citrullinated proteins was also detected at the site of infection with wild-type P. gingivalis. Together, these results suggest bacterial PAD as the mechanistic link between P. gingivalis periodontal infection and rheumatoid arthritis.

Maresz, Katarzyna J.; Hellvard, Annelie; Sroka, Aneta; Adamowicz, Karina; Bielecka, Ewa; Koziel, Joanna; Gawron, Katarzyna; Mizgalska, Danuta; Marcinska, Katarzyna A.; Benedyk, Malgorzata; Pyrc, Krzysztof; Quirke, Anne-Marie; Jonsson, Roland; Alzabin, Saba; Venables, Patrick J.; Nguyen, Ky-Anh

2013-01-01

103

Gingipain-dependent degradation of mammalian target of rapamycin pathway proteins by the periodontal pathogen Porphyromonas gingivalis during invasion.  

Science.gov (United States)

Porphyromonas gingivalis and Tannerella forsythia are gram-negative pathogens strongly associated with periodontitis. Their abilities to interact, invade and persist within host cells are considered crucial to their pathogenicity, but the mechanisms by which they subvert host defences are not well understood. In this study, we set out to investigate whether P. gingivalis and T. forsythia directly target key signalling molecules that may modulate the host cell phenotype to favour invasion and persistence. Our data identify, for the first time, that P. gingivalis, but not T. forsythia, reduces levels of intracellular mammalian target of rapamycin (mTOR) in oral epithelial cells following invasion over a 4-h time course, via the action of gingipains. The ability of cytochalasin D to abrogate P. gingivalis-mediated mTOR degradation suggests that this effect is dependent upon cellular invasion. We also show that levels of several other proteins in the mTOR signalling pathway are modulated by gingipains, either directly or as a consequence of mTOR degradation including p-4E-BP1. Taken together, our data suggest that P. gingivalis manipulates the mTOR pathway, providing evidence for a potentially novel mechanism by which P. gingivalis mediates its effects on host cell responses to infection. PMID:23714361

Stafford, P; Higham, J; Pinnock, A; Murdoch, C; Douglas, C W I; Stafford, G P; Lambert, D W

2013-05-29

104

Gingipain-dependent degradation of mammalian target of rapamycin pathway proteins by the periodontal pathogen Porphyromonas gingivalis during invasion.  

UK PubMed Central (United Kingdom)

Porphyromonas gingivalis and Tannerella forsythia are gram-negative pathogens strongly associated with periodontitis. Their abilities to interact, invade and persist within host cells are considered crucial to their pathogenicity, but the mechanisms by which they subvert host defences are not well understood. In this study, we set out to investigate whether P. gingivalis and T. forsythia directly target key signalling molecules that may modulate the host cell phenotype to favour invasion and persistence. Our data identify, for the first time, that P. gingivalis, but not T. forsythia, reduces levels of intracellular mammalian target of rapamycin (mTOR) in oral epithelial cells following invasion over a 4-h time course, via the action of gingipains. The ability of cytochalasin D to abrogate P. gingivalis-mediated mTOR degradation suggests that this effect is dependent upon cellular invasion. We also show that levels of several other proteins in the mTOR signalling pathway are modulated by gingipains, either directly or as a consequence of mTOR degradation including p-4E-BP1. Taken together, our data suggest that P. gingivalis manipulates the mTOR pathway, providing evidence for a potentially novel mechanism by which P. gingivalis mediates its effects on host cell responses to infection.

Stafford P; Higham J; Pinnock A; Murdoch C; Douglas CW; Stafford GP; Lambert DW

2013-10-01

105

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

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Full Text Available Abstract Background Porphyromonas gingivalis is a Gram-negative intracellular pathogen associated with periodontal disease. We have previously reported on whole-cell quantitative proteomic analyses to investigate the differential expression of virulence factors as the organism transitions from an extracellular to intracellular lifestyle. The original results with the invasive strain P. gingivalis ATCC 33277 were obtained using the genome sequence available at the time, strain W83 [GenBank: AE015924]. We present here a re-processed dataset using the recently published genome annotation specific for strain ATCC 33277 [GenBank: AP009380] and an analysis of differential abundance based on metabolic pathways rather than individual proteins. Results Qualitative detection was observed for 1266 proteins using the strain ATCC 33277 annotation for 18 hour internalized P. gingivalis within human gingival epithelial cells and controls exposed to gingival cell culture medium, an improvement of 7% over the W83 annotation. Internalized cells showed increased abundance of proteins in the energy pathway from asparagine/aspartate amino acids to ATP. The pathway producing one short chain fatty acid, propionate, showed increased abundance, while that of another, butyrate, trended towards decreased abundance. The translational machinery, including ribosomal proteins and tRNA synthetases, showed a significant increase in protein relative abundance, as did proteins responsible for transcription. Conclusion Use of the ATCC 33277 specific genome annotation resulted in improved proteome coverage with respect to the number of proteins observed both qualitatively in terms of protein identifications and quantitatively in terms of the number of calculated abundance ratios. Pathway analysis showed a significant increase in overall protein synthetic and transcriptional machinery in the absence of significant growth. These results suggest that the interior of host cells provides a more energy rich environment compared to the extracellular milieu. Shifts in the production of cytotoxic fatty acids by intracellular P. gingivalis may play a role in virulence. Moreover, despite extensive genomic re-arrangements between strains W83 and 33277, there is sufficient sequence similarity at the peptide level for proteomic abundance trends to be largely accurate when using the heterologous strain annotated genome as the reference for database searching.

Hendrickson Erik L; Xia Qiangwei; Wang Tiansong; Lamont Richard J; Hackett Murray

2009-01-01

106

Virulencia y variabilidad de Porphyromonas gingivalis y Aggregatibacter actinomycetemcomitans y su asociación a la periodontitis/ Virulence and variability on Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans and their association to periodontitis  

Scientific Electronic Library Online (English)

Full Text Available Abstract in spanish Las periodontitis son un conjunto de patologías de naturaleza inflamatoria y etiología infecciosa producidas por el biofilm patogénico subgingival. Porphyromonas gingivalis y Aggregatibacter actinomycetemcomitans son bacterias periodonto-patógenas que pueden causar daño directo a las estructuras periodontales a través de los diversos factores de virulencia que expresan. Sobre la base de estos factores de virulencia, distintos genotipos y serotipos bacterianos se han (more) descrito, cada uno de ellos con una potencial variable patogenicidad. En esta revisión bibliográfica se describen diferentes factores de virulencia de P. gingivalis y A. actinomycetemcomitans y se discute la variable inmunogenicidad y patogenicidad de los distintos genotipos y serotipos descritos para ellos. Tanto P. gingivalis como A. actinomycetemcomitans poseen diversos factores de virulencia asociados al inicio, progresión y severidad de las periodontitis. En P. gingivalis, los factores de virulencia para los cuales se describen distintos genotipos y/o serotipos son fimbria, LPS y cápsula bacteriana, y en A. actinomycetemcomitans son leucotoxina A, Cdt y LPS. Cada uno de estos distintos genotipos y serotipos induce una respuesta inmuno-inflamatoria diferente en el hospedero y, por lo tanto, se podrían asociar a una variable patogenicidad y podrían determinar las características clínicas de la enfermedad. Abstract in english Periodontitis represents a heterogenic group of periodontal infections elicited by bacteria residing at the subgingival biofilm. Although this biofilm is constituted by a broad variety of bacterial species, only a limited number has been associated with the periodontitis aetiology, among them Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans. Both P. gingivalis and A. actinomycetemcomitans express a number of virulence factors that contribute to direct ti (more) ssue damage and, based on them, distinct genotypes and serotypes have been described, each one with a potential variable pathogenicity. This review aimed to analyze the different virulence factors described for P. gingivalis and A. actinomycetemcomitans and to discuss the variable immunogenicity and pathogenicity of their serotypes and genotypes. P. gingivalis and A. actinomycetemcomitans express different virulence factors and they determine the initiation, progression, and severity of periodontitis. In P. gingivalis, distinct serotypes and/or genotypes are described based on fimbriae, LPS, and capsule. Additionally, in A. actinomycetemcomitans distinct serotypes and/or genotypes are described based on leucotoxin A, Cdt, and LPS. These distinct serotypes and genotypes induce a differential immunoinflammatory response and, thus, could be associated with variations in pathogenicity and reflected in clinic characteristics of the disease.

Díaz Zúñiga, J; Yáñez Figueroa, J; Melgar Rodríguez, S; Álvarez Rivas, C; Rojas Lagos, C; Vernal Astudillo, R

2012-04-01

107

Periodontal Treatment Decreases Levels of Antibodies to Porphyromonas Gingivalis and Citrulline in Patients With Rheumatoid Arthritis and Periodontitis.  

UK PubMed Central (United Kingdom)

Background: Porphyromonas gingivalis has been implicated as an etiological agent of rheumatoid arthritis (RA) due to the expression of peptidylarginine deiminase. The present study was undertaken to evaluate whether periodontal treatment may affect serum antibodies to P. gingivalis and citrulline levels in relation to disease activity of RA. Methods: Fifty-five patients with RA were randomly assigned to receive oral hygiene instruction and supragingival scaling (treatment group, n = 26) or no periodontal treatment (control group, n = 29). Periodontal and rheumatologic parameters and serum levels of cytokine and inflammatory markers, citrulline and immunoglobulin G (IgG) to P. gingivalis were examined at baseline and 8 weeks later. Results: Both groups did not differ statistically in all parameters except for % sites with probing depth and clinical attachment level ? 4 mm at baseline. The treatment group exhibited a significantly greater decrease in disease activity score including 28 joints using C-reactive protein (DAS28-CRP) (P = 0.02), serum levels of IgG to P. gingivalis hemin binding protein 35 (HBP35) (P = 0.04) and citrulline (P = 0.02) than the control group. Serum levels of IgG to P. gingivalis HBP35 were significantly correlated positively with those of anti-CCP antibodies (P = 0.0002). The same correlation was obtained between serum levels of IgG to P. gingivalis sonicated extracts and those of rheumatoid factor (P = 0.02) Conclusions: These results suggest that supragingival scaling decreases DAS28-CRP and serum levels of IgG to P. gingivalis HBP35 and citrulline in RA patients. These observations may reflect a role of P. gingivalis in the protein citrullination, which is related to the pathogenesis of RA.

Okada M; Kobayashi T; Ito S; Yokoyama T; Abe A; Murasawa A; Yoshie H

2013-05-01

108

Periodontal Treatment Decreases Levels of Antibodies to Porphyromonas Gingivalis and Citrulline in Patients With Rheumatoid Arthritis and Periodontitis.  

Science.gov (United States)

Background: Porphyromonas gingivalis has been implicated as an etiological agent of rheumatoid arthritis (RA) due to the expression of peptidylarginine deiminase. The present study was undertaken to evaluate whether periodontal treatment may affect serum antibodies to P. gingivalis and citrulline levels in relation to disease activity of RA. Methods: Fifty-five patients with RA were randomly assigned to receive oral hygiene instruction and supragingival scaling (treatment group, n = 26) or no periodontal treatment (control group, n = 29). Periodontal and rheumatologic parameters and serum levels of cytokine and inflammatory markers, citrulline and immunoglobulin G (IgG) to P. gingivalis were examined at baseline and 8 weeks later. Results: Both groups did not differ statistically in all parameters except for % sites with probing depth and clinical attachment level ? 4 mm at baseline. The treatment group exhibited a significantly greater decrease in disease activity score including 28 joints using C-reactive protein (DAS28-CRP) (P = 0.02), serum levels of IgG to P. gingivalis hemin binding protein 35 (HBP35) (P = 0.04) and citrulline (P = 0.02) than the control group. Serum levels of IgG to P. gingivalis HBP35 were significantly correlated positively with those of anti-CCP antibodies (P = 0.0002). The same correlation was obtained between serum levels of IgG to P. gingivalis sonicated extracts and those of rheumatoid factor (P = 0.02) Conclusions: These results suggest that supragingival scaling decreases DAS28-CRP and serum levels of IgG to P. gingivalis HBP35 and citrulline in RA patients. These observations may reflect a role of P. gingivalis in the protein citrullination, which is related to the pathogenesis of RA. PMID:23701010

Okada, Moe; Kobayashi, Tetsuo; Ito, Satoshi; Yokoyama, Tomoko; Abe, Asami; Murasawa, Akira; Yoshie, Hiromasa

2013-05-23

109

[FimA fimbriae of the periodontal disease-associated bacterium porphyromonas gingivalis].  

UK PubMed Central (United Kingdom)

The periodontal disease-associated bacterium Porphyromonas gingivalis primarily uses FimA fimbriae for adhesion to and colonization in the gingival tissues. The fimbriae show a filamentous structure that is composed of polymer of FimA encoded by the fimA gene. FimC, FimD and FimE are associated with the fimbriae as minor components. FimB anchors the fimbriae to the bacterial surface and regulates their length. The N terminus of FimA is digested in a maturation process, then mature FimA proteins are polymerized to form fimbriae in the outer membrane. Transcription of the fimA gene is regulated by the two-component regulatory system of FimS/R. In addition, expression of FimA is influenced by many environmental factors such as nutrients, environmental stresses, and other bacterial products. The fimA gene shows a genetic polymorphism and it is proposed that there are six genotypes (types I-V and Ib). Types II and IV are frequently isolated from severe periodontal patients. Therefore, they are predicted to be high virulent types, but the molecular mechanisms remain unclear. FimA fimbriae also exhibit a heterogenic antigenicity that is basically consistent with the fimA genotype. The fimbriae interact with many molecules such as surface molecules of host cells, extracellular matrix, salivary components, and bacterial components. Many reports argue binding of FimA residues in the fimbriae to the target molecules, but it is reported that accessory components of FimCDE critically function as an adhesin. Elucidation of adherent mechanism of P. gingivalis through the FimA fimbriae could lead to a development of prophylaxis against the bacterial infection.

Nagano K

2013-01-01

110

Identification of Porphyromonas gingivalis proteins secreted by the Por secretion system.  

Science.gov (United States)

The Gram-negative bacterium Porphyromonas gingivalis possesses a number of potential virulence factors for periodontopathogenicity. In particular, cysteine proteinases named gingipains are of interest given their abilities to degrade host proteins and process other virulence factors such as fimbriae. Gingipains are translocated on the cell surface or into the extracellular milieu by the Por secretion system (PorSS), which consists of a number of membrane or periplasmic proteins including PorK, PorL, PorM, PorN, PorO, PorP, PorQ, PorT, PorU, PorV (PG27, LptO), PorW and Sov. To identify proteins other than gingipains secreted by the PorSS, we compared the proteomes of P. gingivalis strains kgp rgpA rgpB (PorSS-proficient strain) and kgp rgpA rgpB porK (PorSS-deficient strain) using two-dimensional gel electrophoresis and peptide-mass fingerprinting. Sixteen spots representing 10 different proteins were present in the particle-free culture supernatant of the PorSS-proficient strain but were absent or faint in that of the PorSS-deficient strain. These identified proteins possessed the C-terminal domains (CTDs), which had been suggested to form the CTD protein family. These results indicate that the PorSS is used for secretion of a number of proteins other than gingipains and that the CTDs of the proteins are associated with the PorSS-dependent secretion. PMID:23075153

Sato, Keiko; Yukitake, Hideharu; Narita, Yuka; Shoji, Mikio; Naito, Mariko; Nakayama, Koji

2012-11-08

111

Identification of Porphyromonas gingivalis proteins secreted by the Por secretion system.  

UK PubMed Central (United Kingdom)

The Gram-negative bacterium Porphyromonas gingivalis possesses a number of potential virulence factors for periodontopathogenicity. In particular, cysteine proteinases named gingipains are of interest given their abilities to degrade host proteins and process other virulence factors such as fimbriae. Gingipains are translocated on the cell surface or into the extracellular milieu by the Por secretion system (PorSS), which consists of a number of membrane or periplasmic proteins including PorK, PorL, PorM, PorN, PorO, PorP, PorQ, PorT, PorU, PorV (PG27, LptO), PorW and Sov. To identify proteins other than gingipains secreted by the PorSS, we compared the proteomes of P. gingivalis strains kgp rgpA rgpB (PorSS-proficient strain) and kgp rgpA rgpB porK (PorSS-deficient strain) using two-dimensional gel electrophoresis and peptide-mass fingerprinting. Sixteen spots representing 10 different proteins were present in the particle-free culture supernatant of the PorSS-proficient strain but were absent or faint in that of the PorSS-deficient strain. These identified proteins possessed the C-terminal domains (CTDs), which had been suggested to form the CTD protein family. These results indicate that the PorSS is used for secretion of a number of proteins other than gingipains and that the CTDs of the proteins are associated with the PorSS-dependent secretion.

Sato K; Yukitake H; Narita Y; Shoji M; Naito M; Nakayama K

2013-01-01

112

Role of MyD88-dependent and MyD88-independent signaling in Porphyromonas gingivalis-elicited macrophage foam cell formation.  

UK PubMed Central (United Kingdom)

Clinical studies and experimental modeling identify a potential link between periodontal disease and periodontal pathogens such as Porphyromonas gingivalis and atherosclerosis and formation of macrophage foam cells. Toll-like receptors and molecules governing their intracellular signaling pathways such as MyD88 play roles in atherosclerosis, as well as host response to P. gingivalis. The aim of this study was to define roles of MyD88 and TRIF during macrophage foam cell formation in response to P. gingivalis. In the presence of human low-density lipoprotein (LDL) mouse bone-marrow-derived macrophages (BM?) cultured with P. gingivalis responded with significant reduction in tumor necrosis factor-? (TNF-?) and interleukin-6 (IL-6). The BM? stained strongly with oil red O, regardless of whether bacterial challenge occurred concurrent with or before LDL treatment. Heat-killed P. gingivalis stimulated foam cell formation in a similar way to live bacteria. The BM? from MyD88-knockout and Lps2 mice revealed a significant role for MyD88, and a minor role for TRIF in P. gingivalis-elicited foam cell formation. Porphyromonas gingivalis-elicited TNF-? and IL-6 were affected by MyD88 ablation and to a lesser extent by TRIF status. These data indicate that LDL affects the TNF-? and IL-6 response of macrophages to P. gingivalis challenge and that MyD88 and TRIF play important roles in P. gingivalis-elicited foam cell formation.

Shaik-Dasthagirisaheb YB; Huang N; Baer MT; Gibson FC 3rd

2013-02-01

113

Species specificity, surface exposure, protein expression, immunogenicity, and participation in biofilm formation of Porphyromonas gingivalis HmuY.  

UK PubMed Central (United Kingdom)

BACKGROUND: Porphyromonas gingivalis is a major etiological agent of chronic periodontitis. The aim of this study was to examine the species specificity, surface exposure, protein expression, immunogenicity, and participation in biofilm formation of the P. gingivalis heme-binding protein HmuY. RESULTS: HmuY is a unique protein of P. gingivalis since only low amino-acid sequence homology has been found to proteins encoded in other species. It is exposed on the cell surface and highly abundant in the outer membrane of the cell, in outer-membrane vesicles, and is released into culture medium in a soluble form. The protein is produced constitutively at low levels in bacteria grown under high-iron/heme conditions and at higher levels in bacteria growing under the low-iron/heme conditions typical of dental plaque. HmuY is immunogenic and elicits high IgG antibody titers in rabbits. It is also engaged in homotypic biofilm formation by P. gingivalis. Anti-HmuY antibodies exhibit inhibitory activity against P. gingivalis growth and biofilm formation. CONCLUSIONS: Here it is demonstrated that HmuY may play a significant role not only in heme acquisition, but also in biofilm accumulation on abiotic surfaces. The data also suggest that HmuY, as a surface-exposed protein, would be available for recognition by the immune response during chronic periodontitis and the production of anti-HmuY antibodies may inhibit biofilm formation.

Olczak T; Wójtowicz H; Ciuraszkiewicz J; Olczak M

2010-01-01

114

Inhibition of Urokinase-Type Plasminogen Activator Expression by Macelignan in Porphyromonas gingivalis Supernatant-Induced Human Oral Epithelial Cells  

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Full Text Available This study was to investigate the effect of macelignan on Porphyromonas gingivalis supernatant-induced uPA expression via regulating mitogen-activated protein kinase (MAPK) and activating protein-1 (AP-1) signaling pathways in human oral epithelial KB cells using casein zymography, Western blotting, reverse transcription-PCR and reporter gene assays. Zymographic analysis of secreted enzymes identified the main caseinolytic band at 54 kDa. Macelignan inhibited the expression of uPA protein and mRNA, as well uPA secretion, in KB cells exposed to P. gingivalis supernatant. Consistent with these findings, macelignan suppressed phosphorylation of p38 and c-Jun N terminal kinase (JNK) in P. gingivalis supernatant-induced KB cells. The levels of c-Fos and phosphorylated c-Jun, which together form AP-1, the transcription factor that is involved in uPA gene expression, were partially reduced by macelignan. Macelignan also blocked P. gingivalis supernatant-induced AP-1 activity in these cells. These results suggest that macelignan decreased P. gingivalis supernatant-induced uPA expression by blocking AP-1 activity, which may be mediated by inhibition of phosphorylation of p38 and JNK in KB cells. Macelignan may potently use for the modulation of periodontal inflammation.

YANTI

2010-01-01

115

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

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Full Text Available Background: Biofilms are organized communities of microorganisms embedded in a self-produced extracellular polymeric matrix (EPM), often with great phylogenetic variety. Bacteria in the subgingival biofilm are key factors that cause periodontal diseases; among these are the Gram-negative bacteria Fusobacterium nucleatum and Porphyromonas gingivalis. The objectives of this study were to characterize the major components of the EPM and to test the effect of deoxyribonuclease I (DNase I) and proteinase K. Methods: F. nucleatum and P. gingivalis bacterial cells were grown in dynamic and static biofilm models. The effects of DNase I and proteinase K enzymes on the major components of the EPM were tested during biofilm formation and on mature biofilm. Confocal laser scanning microscopy was used in observing biofilm structure. Results: Proteins and carbohydrates were the major components of the biofilm matrix, and extracellular DNA (eDNA) was also present. DNase I and proteinase K enzymes had little effect on biofilms in the conditions used. In the flow cell, F. nucleatum was able to grow in partially oxygenated conditions while P. gingivalis failed to form biofilm alone in similar conditions. F. nucleatum supported the growth of P. gingivalis when they were grown together as dual species biofilm. Conclusion: DNase I and proteinase K had little effect on the biofilm matrix in the conditions used. F. nucleatum formed biofilm easily and supported the growth of P. gingivalis, which preferred anaerobic conditions.

Marwan Mansoor Ali Mohammed; Audun H. Nerland; Mohammed Al-Haroni; Vidar Bakken

2013-01-01

116

Variabilidad de la síntesis de RANKL por linfocitos T frente a distintos serotipos capsulares de Porphyromonas gingivalis Variability in the RANKL synthesis by T lymphocytes in response to different Porphyromonas gingivalis capsular serotypes  

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Full Text Available Propósito: Las periodontitis representan un grupo heterogéneo de infecciones periodontales cuya etiología son las bacterias residentes en el biofilm subgingival. Aunque este biofilm está constituido por una amplia variedad de especies bacterianas, sólo un número limitado de especies, como Porphyromonas gingivalis, se ha asociado a la etiología de la enfermedad. P. gingivalis expresa diversos factores de virulencia que pueden causar daño directo a los tejidos del hospedero; sin embargo, su mayor patogenicidad involucra la inducción de una respuesta inmuno-inflamatoria, durante la cual se secretan una amplia variedad de citoquinas, quimioquinas y mediadores inflamatorios que pueden inducir la destrucción de los tejidos de soporte de los dientes y la pérdida de ellos. Método: En esta investigación, se evaluó si los distintos serotipos capsulares (K) de P. gingivalis pueden determinar los niveles de síntesis de RANKL, citoquina clave en la destrucción del hueso alveolar durante la periodontitis. Para ello, se cuantificaron los niveles de expresión de RANKL mediante PCR cuantitativa y los niveles de secreción mediante ELISA en linfocitos T activados en presencia de los serotipos capsulares K1-K6 de P. gingivalis, y estos se correlacionaron a los niveles de expresión de los factores de transcripción asociados a cada uno de los fenotipos de linfocitos efectores: Th1 (T-bet), Th2 (GATA-3), Th17 (RORC2) y Treg (Foxp3). Resultados: Mayores niveles de expresión y secreción de RANKL fueron detectados en linfocitos T activados en presencia de los serotipos K1 y K2 de P. gingivalis, en comparación a los detectados ante los otros serotipos. Además, estos mayores niveles de RANKL se correlacionaron positivamente con los niveles de expresión de RORC2. Conclusión: Estos datos demuestran que la síntesis de RANKL por linfocitos T se restringe a ciertos serotipos capsulares de P. gingivalis (K1 y K2) y permiten sugerir que los serotipos K1 y K2 de P. gingivalis podrían asociarse a la destrucción del hueso alveolar y a la pérdida de los dientes durante la periodontitis.Aim: Periodontitis represents a heterogenic group of periodontal infections elicited by bacteria residing at the subgingival biofilm. Although this biofilm is constituted by a broad variety of bacterial species, only a limited number has been associated with the periodontitis aetiology, among them Porphyromonas gingivalis. P. gingivalis express a number of virulence factors that contribute to direct tissue damage; however, their pathogenicity relies mainly on the induction of a host immuno-inflammatory response. This leads to the release of a broad array of cytokines, chemokines and inflammatory mediators, which cause destruction of the tooth-supporting alveolar bone and ultimately tooth loss. Method: In the present investigation, in order to determine whether different P. gingivalis serotypes might lead to a differential RANKL synthesis, a key cytokine involved in alveolar bone resorption, the mRNA expression and secretion of RANKL and the expression of transcription factors T-bet, GATA-3, RORC2 and Foxp3, the master-switch genes controlling the Th1, Th2, Th17, and Treg cell differentiation, respectively, were analyzed on human T cells activated with different P. gingivalis capsular (K) serotypes. Results: T lymphocytes responding to P. gingivalis serotypes K1 or K2, but not to the other serotypes, led to an increased expression and secretion of RANKL. In addition, these higher RANKL levels correlate with RORC2 expression upon activation with K1 or K2 serotypes. Conclusion: These data demonstrated that RANKL expression and secretion by T lymphocytes was restricted to particular P. gingivalis serotypes (namely K1 and K2), and allowed to suggest a link between these serotypes with alveolar bone destruction and teeth loosening during the periodontitis.

M Navarrete; A Silva; M Sanz; R Vernal

2010-01-01

117

Variabilidad de la síntesis de RANKL por linfocitos T frente a distintos serotipos capsulares de Porphyromonas gingivalis/ Variability in the RANKL synthesis by T lymphocytes in response to different Porphyromonas gingivalis capsular serotypes  

Scientific Electronic Library Online (English)

Full Text Available Abstract in spanish Propósito: Las periodontitis representan un grupo heterogéneo de infecciones periodontales cuya etiología son las bacterias residentes en el biofilm subgingival. Aunque este biofilm está constituido por una amplia variedad de especies bacterianas, sólo un número limitado de especies, como Porphyromonas gingivalis, se ha asociado a la etiología de la enfermedad. P. gingivalis expresa diversos factores de virulencia que pueden causar daño directo a los tejidos del h (more) ospedero; sin embargo, su mayor patogenicidad involucra la inducción de una respuesta inmuno-inflamatoria, durante la cual se secretan una amplia variedad de citoquinas, quimioquinas y mediadores inflamatorios que pueden inducir la destrucción de los tejidos de soporte de los dientes y la pérdida de ellos. Método: En esta investigación, se evaluó si los distintos serotipos capsulares (K) de P. gingivalis pueden determinar los niveles de síntesis de RANKL, citoquina clave en la destrucción del hueso alveolar durante la periodontitis. Para ello, se cuantificaron los niveles de expresión de RANKL mediante PCR cuantitativa y los niveles de secreción mediante ELISA en linfocitos T activados en presencia de los serotipos capsulares K1-K6 de P. gingivalis, y estos se correlacionaron a los niveles de expresión de los factores de transcripción asociados a cada uno de los fenotipos de linfocitos efectores: Th1 (T-bet), Th2 (GATA-3), Th17 (RORC2) y Treg (Foxp3). Resultados: Mayores niveles de expresión y secreción de RANKL fueron detectados en linfocitos T activados en presencia de los serotipos K1 y K2 de P. gingivalis, en comparación a los detectados ante los otros serotipos. Además, estos mayores niveles de RANKL se correlacionaron positivamente con los niveles de expresión de RORC2. Conclusión: Estos datos demuestran que la síntesis de RANKL por linfocitos T se restringe a ciertos serotipos capsulares de P. gingivalis (K1 y K2) y permiten sugerir que los serotipos K1 y K2 de P. gingivalis podrían asociarse a la destrucción del hueso alveolar y a la pérdida de los dientes durante la periodontitis. Abstract in english Aim: Periodontitis represents a heterogenic group of periodontal infections elicited by bacteria residing at the subgingival biofilm. Although this biofilm is constituted by a broad variety of bacterial species, only a limited number has been associated with the periodontitis aetiology, among them Porphyromonas gingivalis. P. gingivalis express a number of virulence factors that contribute to direct tissue damage; however, their pathogenicity relies mainly on the inductio (more) n of a host immuno-inflammatory response. This leads to the release of a broad array of cytokines, chemokines and inflammatory mediators, which cause destruction of the tooth-supporting alveolar bone and ultimately tooth loss. Method: In the present investigation, in order to determine whether different P. gingivalis serotypes might lead to a differential RANKL synthesis, a key cytokine involved in alveolar bone resorption, the mRNA expression and secretion of RANKL and the expression of transcription factors T-bet, GATA-3, RORC2 and Foxp3, the master-switch genes controlling the Th1, Th2, Th17, and Treg cell differentiation, respectively, were analyzed on human T cells activated with different P. gingivalis capsular (K) serotypes. Results: T lymphocytes responding to P. gingivalis serotypes K1 or K2, but not to the other serotypes, led to an increased expression and secretion of RANKL. In addition, these higher RANKL levels correlate with RORC2 expression upon activation with K1 or K2 serotypes. Conclusion: These data demonstrated that RANKL expression and secretion by T lymphocytes was restricted to particular P. gingivalis serotypes (namely K1 and K2), and allowed to suggest a link between these serotypes with alveolar bone destruction and teeth loosening during the periodontitis.

Navarrete, M; Silva, A; Sanz, M; Vernal, R

2010-04-01

118

Influence of periodontal disease, Porphyromonas gingivalis and cigarette smoking on systemic anti-citrullinated peptide antibody titres.  

UK PubMed Central (United Kingdom)

BACKGROUND: Anti-citrullinated protein antibody (ACPA) responses may precede clinical onset of rheumatoid arthritis. Porphyromonas gingivalis peptidylarginine deiminase can citrullinate proteins possibly inducing autoimmunity in susceptible individuals. AIM: To determine whether periodontitis, carriage of P. gingivalis, smoking and periodontal therapy influence ACPA titres. METHODS: Serum and plaque samples were collected from 39 periodontitis patients before and after non-surgical periodontal treatment, and from 36 healthy subjects. Carriage of P. gingivalis was determined by PCR of plaque DNA. ACPA was determined by anti-cyclic citrullinated peptide (CCP) enzyme-linked immunosorbent assay (ELISA). Anti-P. gingivalis titres were determined by ELISA. RESULTS: Untreated periodontitis patients had higher anti-CCP antibody titres than healthy controls [three patients (8%) greater than manufacturer suggested assay diagnostic threshold (5 Assay Units/AU) versus none (0%); mean ± SEM: 1.37 ± 0.23 versus 0.40 ± 0.10 AU, p < 0.0001]. Periodontitis patients who smoked demonstrated lower anti-P. gingivalis (15956 ± 4385 versus 2512 ± 1290 Units/ml, p < 0.05), but similar anti-CCP than non-smoking periodontitis patients (smokers: 1.31 ± 0.35; non-smokers: 1.41 ± 0.32 AU). Healthy smokers demonstrated elevated anti-CCP titres (0.75 ± 0.19 AU), at levels between healthy non-smokers (0.15 ± 0.05 AU) and non-smoker periodontitis patients. Six months after periodontal treatment, there were significant reductions in anti-CCP (non-smokers p < 0.05) and anti-P. gingivalis (all participants p < 0.01). CONCLUSION: In subjects with periodontitis, P. gingivalis infection may be responsible for inducing autoimmune responses that characterize rheumatoid arthritis.

Lappin DF; Apatzidou D; Quirke AM; Oliver-Bell J; Butcher JP; Kinane DF; Riggio MP; Venables P; McInnes IB; Culshaw S

2013-10-01

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Survival in transport media of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Prevotella intermedia in human subgingival samples.  

UK PubMed Central (United Kingdom)

The purpose of this study was to evaluate the survival in 3 transport media of 3 suspected periodontal pathogens, Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Prevotella intermedia. Subgingival samples were taken from 10 patients with severe periodontitis, all harboring at least two of the above-mentioned species. The material was dispersed and aliquots were added to vials containing reduced transport fluid, reduced transport fluid containing 10% Fildes extract, or viability-maintaining microbiostatic medium, anaerobically prepared (VMGA III). Viable counts were determined after 1, 2, 4, 24 and 48 h of storage at 4 degrees C or at room temperature. The results showed that, for up to 4 h of storage, no significant differences existed for all parameters tested. A large increase of the total viable counts was found in VMGA III at room temperature after 24 and 48 h. This was due to an outgrowth of mainly streptococci. Incubation at 4 degrees C yielded often a significantly higher recovery compared to room temperature. After storage at room temperature, the tested bacteria were below detection level in some samples.

van Steenbergen TJ; Petit MD; Tijhof CJ; van Winkelhoff AJ; van der Velden U; de Graaff J

1993-12-01

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Survival in transport media of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Prevotella intermedia in human subgingival samples.  

Science.gov (United States)

The purpose of this study was to evaluate the survival in 3 transport media of 3 suspected periodontal pathogens, Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Prevotella intermedia. Subgingival samples were taken from 10 patients with severe periodontitis, all harboring at least two of the above-mentioned species. The material was dispersed and aliquots were added to vials containing reduced transport fluid, reduced transport fluid containing 10% Fildes extract, or viability-maintaining microbiostatic medium, anaerobically prepared (VMGA III). Viable counts were determined after 1, 2, 4, 24 and 48 h of storage at 4 degrees C or at room temperature. The results showed that, for up to 4 h of storage, no significant differences existed for all parameters tested. A large increase of the total viable counts was found in VMGA III at room temperature after 24 and 48 h. This was due to an outgrowth of mainly streptococci. Incubation at 4 degrees C yielded often a significantly higher recovery compared to room temperature. After storage at room temperature, the tested bacteria were below detection level in some samples. PMID:8152838

van Steenbergen, T J; Petit, M D; Tijhof, C J; van Winkelhoff, A J; van der Velden, U; de Graaff, J

1993-12-01

 
 
 
 
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Crystal structure and mechanism of tripeptidyl activity of prolyl tripeptidyl aminopeptidase from Porphyromonas gingivalis.  

Science.gov (United States)

The crystal structure of prolyl tripeptidyl aminopeptidase from Porphyromonas gingivalis was determined. Prolyl tripeptidyl aminopeptidase consists of beta-propeller and catalytic domains, and a large cavity between the domains; this structure is similar to dipeptidyl aminopeptidase IV. A catalytic triad (Ser603, His710, and Asp678) was located in the catalytic domain; this triad was virtually identical to that of the enzymes belonging to the prolyl oligopeptidase family. The structure of an inactive S603A mutant enzyme complexed with a substrate was also determined. The pyrrolidine ring of the proline residue appeared to fit into a hydrophobic pocket composed of Tyr604, Val629, Trp632, Tyr635, Tyr639, Val680, and Val681. There were characteristic differences in the residues of the beta-propeller domain, and these differences were related to the substrate specificity of tripeptidyl activity. The N-terminal amino group was recognized by salt bridges, with two carboxyl groups of Glu205 and Glu206 from a helix in dipeptidyl aminopeptidase IV. In prolyl tripeptidyl aminopeptidase, however, the Glu205 (located in the loop) and Glu636 were found to carry out this function. The loop structure provides sufficient space to accommodate three N-terminal residues (Xaa-Xaa-Pro) of substrates. This is the first report of the structure and substrate recognition mechanism of tripeptidyl peptidase. PMID:16914159

Ito, Kiyoshi; Nakajima, Yoshitaka; Xu, Yue; Yamada, Nozomi; Onohara, Yuko; Ito, Takashi; Matsubara, Futoshi; Kabashima, Tsutomu; Nakayama, Koji; Yoshimoto, Tadashi

2006-08-17

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Crystal structure and mechanism of tripeptidyl activity of prolyl tripeptidyl aminopeptidase from Porphyromonas gingivalis.  

UK PubMed Central (United Kingdom)

The crystal structure of prolyl tripeptidyl aminopeptidase from Porphyromonas gingivalis was determined. Prolyl tripeptidyl aminopeptidase consists of beta-propeller and catalytic domains, and a large cavity between the domains; this structure is similar to dipeptidyl aminopeptidase IV. A catalytic triad (Ser603, His710, and Asp678) was located in the catalytic domain; this triad was virtually identical to that of the enzymes belonging to the prolyl oligopeptidase family. The structure of an inactive S603A mutant enzyme complexed with a substrate was also determined. The pyrrolidine ring of the proline residue appeared to fit into a hydrophobic pocket composed of Tyr604, Val629, Trp632, Tyr635, Tyr639, Val680, and Val681. There were characteristic differences in the residues of the beta-propeller domain, and these differences were related to the substrate specificity of tripeptidyl activity. The N-terminal amino group was recognized by salt bridges, with two carboxyl groups of Glu205 and Glu206 from a helix in dipeptidyl aminopeptidase IV. In prolyl tripeptidyl aminopeptidase, however, the Glu205 (located in the loop) and Glu636 were found to carry out this function. The loop structure provides sufficient space to accommodate three N-terminal residues (Xaa-Xaa-Pro) of substrates. This is the first report of the structure and substrate recognition mechanism of tripeptidyl peptidase.

Ito K; Nakajima Y; Xu Y; Yamada N; Onohara Y; Ito T; Matsubara F; Kabashima T; Nakayama K; Yoshimoto T

2006-09-01

123

Involvement of a periodontal pathogen, Porphyromonas gingivalis on the pathogenesis of non-alcoholic fatty liver disease  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Non-alcoholic fatty liver disease (NAFLD) is a hepatic manifestation of metabolic syndrome that is closely associated with multiple factors such as obesity, hyperlipidemia and type 2 diabetes mellitus. However, other risk factors for the development of NAFLD are unclear. With the association between periodontal disease and the development of systemic diseases receiving increasing attention recently, we conducted this study to investigate the relationship between NAFLD and infection with Porphyromonas gingivalis (P. gingivalis), a major causative agent of periodontitis. Methods The detection frequencies of periodontal bacteria in oral samples collected from 150 biopsy-proven NAFLD patients (102 with non-alcoholic steatohepatitis (NASH) and 48 with non-alcoholic fatty liver (NAFL) patients) and 60 non-NAFLD control subjects were determined. Detection of P. gingivalis and other periodontopathic bacteria were detected by PCR assay. In addition, effect of P. gingivalis-infection on mouse NAFLD model was investigated. To clarify the exact contribution of P. gingivalis-induced periodontitis, non-surgical periodontal treatments were also undertaken for 3 months in 10 NAFLD patients with periodontitis. Results The detection frequency of P. gingivalis in NAFLD patients was significantly higher than that in the non-NAFLD control subjects (46.7% vs. 21.7%, odds ratio: 3.16). In addition, the detection frequency of P. gingivalis in NASH patients was markedly higher than that in the non-NAFLD subjects (52.0%, odds ratio: 3.91). Most of the P. gingivalis fimbria detected in the NAFLD patients was of invasive genotypes, especially type II (50.0%). Infection of type II P. gingivalis on NAFLD model of mice accelerated the NAFLD progression. The non-surgical periodontal treatments on NAFLD patients carried out for 3 months ameliorated the liver function parameters, such as the serum levels of AST and ALT. Conclusions Infection with high-virulence P. gingivalis might be an additional risk factor for the development/progression of NAFLD/NASH.

Yoneda Masato; Naka Shuhei; Nakano Kazuhiko; Wada Koichiro; Endo Hiroki; Mawatari Hironori; Imajo Kento; Nomura Ryota; Hokamura Kazuya; Ono Masafumi; Murata Shogo; Tohnai Iwai; Sumida Yoshio; Shima Toshihide; Kuboniwa Masae; Umemura Kazuo; Kamisaki Yoshinori; Amano Atsuo; Okanoue Takeshi; Ooshima Takashi; Nakajima Atsushi

2012-01-01

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Vitamin D inhibits the expression of interleukin-8 in human periodontal ligament cells stimulated with Porphyromonas gingivalis.  

UK PubMed Central (United Kingdom)

OBJECTIVE: Vitamin D has been known to be closely associated with periodontitis while the exact mechanisms remain unclear. The present study aimed to discover the effects of 1a,25-dihydroxyvitamin D3 (1,25D) on the expressions of interleukin (IL)-6 and IL-8 in human periodontal ligament cells (hPDLCs) stimulated with Porphyromonas gingivalis (P. gingivalis) W83. DESIGN: Primary cultures of hPDLCs from ten donors were established and the cells of passage four were treated with 1,25D or P. gingivalis individually or 1,25D combined with P. gingivalis. The levels of IL-6 and IL-8 protein in hPDLCs were detected with enzyme-linked immunosorbent assay (ELISA) and the mRNA levels were detected with real-time RT-PCR. RESULTS: P. gingivalis significantly promoted the protein expressions of IL-6 and IL-8. P. gingivalis at the multiplicity of infection (MOI) 100 exerted the strongest promotion effect on the IL-6 protein expression by 5.83-fold compared with the controls (2482.88±26.53pg/ml versus 425.80±77.25pg/ml, P<0.0005) and the IL-8 protein expression by 12.39-fold (4965.81±1072.55pg/ml versus 400.75±2.27pg/ml, P=0.005) in hPDLCs at 24h. At 48h, 10(-8)mol/L 1,25D had the best inhibition on the IL-8 protein expression in hPDLCs by 2.00-fold compared with the controls (100.76±21.11pg/ml versus 201.75±18.15pg/ml, P<0.0005) and the IL-8 mRNA expression by 2.13-fold (P<0.0005). 10(-8)mol/L 1,25D combined with P. gingivalis (MOI 100) exerted the strongest inhibition effect on the IL-8 protein expression by 1.54-fold compared with P. gingivalis treatment alone (3077.33±210.04pg/ml versus 4738.24±1386.17pg/ml, P=0.018) and the IL-8 mRNA expression by 1.78-fold (P=0.012) in hPDLCs at 12h. 1,25D did not influence the expression of IL-6 in hPDLCs with or without P. gingivalis treatment. CONCLUSION: Vitamin D may potentially inhibit the periodontal inflammation induced by P. gingivalis partly by decreasing the IL-8 expression in hPDLCs.

Tang X; Pan Y; Zhao Y

2013-04-01

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Prevalence of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Tannerella forsythia in Japanese patients with generalized chronic and aggressive periodontitis.  

Science.gov (United States)

This study aimed to investigate the prevalence and levels of major periodontal pathogens, Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis and Tannerella forsythia in subgingival plaque samples of a group of Japanese patients with aggressive periodontitis (AgP) and chronic periodontitis (CP). A total of 40 patients with clinical diagnosis of AgP or CP and 10 periodontally healthy volunteers were subjected to clinical and microbiological analysis. Subgingival plaque samples were analyzed for A. actinomycetemcomitans, P. gingivalis and T. forsythia with a real-time polymerase chain reaction (PCR) technique. The prevalence of P. gingivalis and T. forsythia was relatively high in patients with periodontitis: over 60% of AgP or CP patients harbored these pathogens whereas they were not detected in the subgingival plaque samples from periodontally healthy individuals. P. gingivalis and T. forsythia were relatively frequently detected together in AgP and CP patients. No significant differences in the prevalence or level of the 3 pathogens were found between periodontitis groups. The proportion of T. forsythia was approximately 4-fold higher in CP group than in AgP group (P = 0.02). In periodontitis patients, a significant positive correlation was found between periodontal parameters (probing depth and clinical attachment level) and the numbers of total bacteria, P. gingivalis and T. forsythia. No distinct pattern of the subgingival profile of these pathogens was discerned between the two disease entities, except for the difference in the proportion of T. forsythia. The red complex bacteria, P. gingivalis and T. forsythia were highly prevalent in this population of Japanese AgP and CP patients, collaborating their roles in periodontitis. PMID:23608307

Tomita, Sachiyo; Komiya-Ito, Akiyo; Imamura, Kentaro; Kita, Daichi; Ota, Koki; Takayama, Saori; Makino-Oi, Asako; Kinumatsu, Takashi; Ota, Mikio; Saito, Atsushi

2013-04-20

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Porphyromonas gingivalis induces CCR5-dependent transfer of infectious HIV-1 from oral keratinocytes to permissive cells  

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Full Text Available Abstract Background Systemic infection with HIV occurs infrequently through the oral route. The frequency of occurrence may be increased by concomitant bacterial infection of the oral tissues, since co-infection and inflammation of some cell types increases HIV-1 replication. A putative periodontal pathogen, Porphyromonas gingivalis selectively up-regulates expression of the HIV-1 coreceptor CCR5 on oral keratinocytes. We, therefore, hypothesized that P. gingivalis modulates the outcome of HIV infection in oral epithelial cells. Results Oral and tonsil epithelial cells were pre-incubated with P. gingivalis, and inoculated with either an X4- or R5-type HIV-1. Between 6 and 48 hours post-inoculation, P. gingivalis selectively increased the infectivity of R5-tropic HIV-1 from oral and tonsil keratinocytes; infectivity of X4-tropic HIV-1 remained unchanged. Oral keratinocytes appeared to harbor infectious HIV-1, with no evidence of productive infection. HIV-1 was harbored at highest levels during the first 6 hours after HIV exposure and decreased to barely detectable levels at 48 hours. HIV did not appear to co-localize with P. gingivalis, which increased selective R5-tropic HIV-1 trans infection from keratinocytes to permissive cells. When CCR5 was selectively blocked, HIV-1 trans infection was reduced. Conclusion P. gingivalis up-regulation of CCR5 increases trans infection of harbored R5-tropic HIV-1 from oral keratinocytes to permissive cells. Oral infections such as periodontitis may, therefore, increase risk for oral infection and dissemination of R5-tropic HIV-1.

Giacaman Rodrigo A; Asrani Anil C; Gebhard Kristin H; Dietrich Elizabeth A; Vacharaksa Anjalee; Ross Karen F; Herzberg Mark C

2008-01-01

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Effect of Porphyromonas gingivalis ATCC 33277 vesicle on adherence of Streptococcus mutans OMZ 70 to the experimental pellicle.  

UK PubMed Central (United Kingdom)

The vesicles of Porphyromonas gingivalis ATCC 33277 strongly aggregated Streptococcus cricetus, S. rattus, and S. mutans, but poorly aggregated S. sobrinus. The adherence of S. mutans OMZ 70 to hydroxyapatite (HA) coated with whole saliva was increased in parallel with the quantity of the vesicles. The significant increase of adherence of S. mutans OMZ 70 by the vesicles was also observed on the HA coated with parotid saliva, submandibular saliva, serum, and type I collagen. These findings suggest that the vesicles may act as a bridge between mutans streptococcus and the tooth surface.

Kamaguchi A; Baba H; Hoshi M; Inomata K

1995-01-01

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Characterization of Heat-Inducible Expression and Cloning of HtpG (Hsp90 Homologue) of Porphyromonas gingivalis  

Science.gov (United States)

Porphyromonas gingivalis is implicated in the etiology of periodontal disease. Associations between microbial virulence and stress protein expression have been identified in other infections. For example, Hsp90 homologues in several microbial species have been shown to contribute to virulence. We previously reported that P. gingivalis possessed an Hsp90 homologue (HtpG) which cross-reacts with human Hsp90. In addition, we found that elevated levels of serum antibody to Hsp90 stress protein in individuals colonized with this microorganism were associated with periodontal health. However, the role of HtpG in P. gingivalis has not been explored. Therefore, we cloned the htpG gene and investigated the characteristics of HtpG localization and expression in P. gingivalis. htpG exists as a single gene of 2,052 bp from which a single message encoding a mature protein of approximately 68 kDa is transcribed. Western blot analysis revealed that the 68-kDa polypeptide was stress inducible and that a major band at 44 kDa and a minor band at 40 kDa were present at constitutive levels. Cellular localization studies revealed that the 44- and 40-kDa species were associated with membrane and vesicle fractions, while the 68-kDa polypeptide was localized to the cytosolic fractions.

Lopatin, Dennis E.; Combs, Allison; Sweier, Domenica G.; Fenno, J. Christopher; Dhamija, Sangeeta

2000-01-01

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Fucoidan interferes with Porphyromonas gingivalis-induced aneurysm enlargement by decreasing neutrophil activation.  

UK PubMed Central (United Kingdom)

PURPOSE: Neutrophils have been shown to be involved in all stages of human and experimental abdominal aortic aneurysm (AAA) development. The initial processes of neutrophil rolling and trapping in the intraluminal thrombus (ILT) are mediated mainly by P-selectin expressed by activated platelets. In the present study, we propose to evaluate the beneficial effect of fucoidan, a competitive binding agent of P-selectin, on aneurysmal growth in a rat model of aortic aneurysm with neutrophil enrichment of the ILT induced by repeated episodes of weak bacteremia. METHODS: Sixty Lewis rats with experimental AAAs, developed from decellularized aortic xenografts, were divided into four groups. Two groups were used as controls: group fucoidan control (FC) was treated with 200 mg of fucoidan (F) delivered by 2 mL, 4-week osmotic pumps placed intraperitoneally before closing the abdomen, and group C received saline instead of fucoidan. Two more groups were injected weekly with Porphyromonas gingivalis (P. gingivalis [Pg]): group F+Pg received 200 mg of intraperitoneal fucoidan and group Pg received saline. AAAs were harvested after 4 weeks and peripheral blood was sampled at that time. Cell-free DNA (cf-DNA) and myeloperoxydase (MPO) antigen concentrations were determined in plasma and in AAA-conditioned media. Histology and P-selectin immunostaining were performed on AAA tissue samples. RESULTS: Comparing rats injected with Pg, those receiving fucoidan presented reduced aneurysmal diameter. Histologic analysis of AAAs showed that fucoidan reduced the ILT thickness in Pg-injected rats, with fewer trapped neutrophils, and with signs of a healing process, as observed in control group C. Immunohistological analysis revealed a substantial decrease in P-selectin immunostaining at the luminal surface of aneurysms in fucoidan-treated rats compared to the other groups, suggesting an interaction between fucoidan and P-selectin. A significant decrease in MPO concentrations in both plasma and conditioned medium was induced by fucoidan treatment in Pg-injected rats, reflecting a pacification of the ILT biological activity. This effect was associated with a reduction in neutrophil activation and apoptosis, reflected by a significant decrease in cf-DNA concentration in both plasma and conditioned medium of fucoidan-treated rats. CONCLUSIONS: Our results suggest that fucoidan has a beneficial effect on experimental aneurysmal degeneration by decreasing neutrophil activation in the ILT enhanced by weak pathogen contamination. This effect seems to be related to its interaction with P-selectin, which may decrease the trapping of neutrophils into the ILT. Fucoidan could represent a therapeutic option in AAAs to decrease the neutrophil activation involved in the degenerative process of aneurysmal expansion and rupture.

Alsac JM; Delbosc S; Rouer M; Journé C; Louedec L; Meilhac O; Michel JB

2013-03-01

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Decreased interleukin-2 responses to Fusobacterium nucleatum and Porphyromonas gingivalis in generalized aggressive periodontitis  

DEFF Research Database (Denmark)

BACKGROUND: Compromised T-cell responses to periodontal pathogens may contribute to the pathogenesis of generalized aggressive periodontitis (GAgP). In this study, we attempted to characterize T-helper cell (Th1, Th2, and Th17) responses in patients with GAgP and healthy controls upon stimulation with disease-relevant pathogens. METHODS: Mononuclear cells (MNCs) from 10 white patients with GAgP and 10 white controls were stimulated with Porphyromonas gingivalis American Type Culture Collection (ATCC) 33277 (Pg), Prevotella intermedia ATCC 25611, Fusobacterium nucleatum ATCC 49256 (Fn), and similar bacteria isolated from the participants' inherent oral flora. Tetanus toxoid (TT) was used as control antigen. The resulting production of interferon-gamma (IFN-gamma) and interleukin (IL)-2, -4, -5 and -17 and the induced proliferation of CD4+ T cells were measured. RESULTS: MNCs from patients with GAgP exhibited decreased IL-2 responses to Pg and Fn. No difference was observed between patients with GAgP and controls with regard to CD4+ T-cell proliferation or the production of IFN-gamma and IL-4, -5, and -17, irrespective of whether type strains or bacteria isolated from the participants' oral cavity were used for stimulation. Moreover, similar proliferative and cytokine responses to TT were observed. Notably, smoking patients with GAgP exhibited significantly lower IFN-gamma responses to the bacteria and to TT than non-smoking patients or controls. CONCLUSIONS: The decreased IL-2 responses of patients with GAgP to Pg and Fn combined with adequate IL-2 responses to TT suggest an impaired antigen-specific T-cell reactivity with periodontal pathogens in GAgP. The decreased IFN-gamma responses of smokers within the patient group suggest that smoking may aggravate this impairment.

Borch, Tanja SkuldbØl; Pedersen, Morten LØbner

2009-01-01

131

Regulon controlled by the GppX hybrid two component system in Porphyromonas gingivalis.  

UK PubMed Central (United Kingdom)

The periodontal pathogen Porphyromonas gingivalis experiences a number of environmental conditions in the oral cavity, and must monitor and respond to a variety of environmental cues. However, the organism possesses only five full two-component systems, one of which is the hybrid system GppX. To investigate the regulon controlled by GppX we performed RNA-Seq on a ?GppX mutant. Fifty-three genes were upregulated and 37 genes were downregulated in the ?GppX mutant. Pathway analyses revealed no systemic function for GppX under nutrient-replete conditions; however, over 40% of the differentially abundant genes were annotated as encoding hypothetical proteins indicating a novel role for GppX. Abundance of small RNA was, in general, not affected by the absence of GppX. To further define the role of GppX with respect to regulation of a hypothetical protein observed with the greatest significant relative abundance change relative to a wild-type control, PGN_0151, we constructed a series of strains in which the ?gppX mutation was complemented with a GppX protein containing specific domain and phosphotransfer mutations. The transmembrane domains, the DNA-binding domain and the phosphotransfer residues were all required for regulation of PGN_0151. In addition, binding of GppX to the PGN_0151 promoter regions was confirmed by an electrophoretic mobility shift assay. Both the ?GppX mutant and a ?PGN_0151 mutant were deficient in monospecies biofilm formation, suggesting a role for the GppX-PGN_0151 regulon in colonization and survival of the organism.

Hirano T; Beck DA; Wright CJ; Demuth DR; Hackett M; Lamont RJ

2013-02-01

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Trimeric structure of major outer membrane proteins homologous to OmpA in Porphyromonas gingivalis.  

Science.gov (United States)

The major outer membrane proteins Pgm6 (41 kDa) and Pgm7 (40 kDa) of Porphyromonas gingivalis ATCC 33277 are encoded by open reading frames pg0695 and pg0694, respectively, which form a single operon. Pgm6 and Pgm7 (Pgm6/7) have a high degree of similarity to Escherichia coli OmpA in the C-terminal region and are predicted to form eight-stranded beta-barrels in the N-terminal region. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Pgm6/7 appear as bands with apparent molecular masses of 40 and 120 kDa, with and without a reducing agent, suggesting a monomer and trimer, respectively. To verify the predicted trimeric structure and function of Pgm6/7, we constructed three mutants with pg0695, pg0694, or both deleted. The double mutant produced no Pgm6/7. The single-deletion mutants appeared to contain less Pgm7 and Pgm6 and to form homotrimers that migrated slightly faster (115 kDa) and slower (130 kDa), respectively, than wild-type Pgm6/7 under nonreducing conditions. N-terminal amino acid sequencing and mass spectrometry analysis of partially digested Pgm6/7 detected only fragments from Pgm6 and Pgm7. Two-dimensional, diagonal electrophoresis and chemical cross-linking experiments with or without a reducing agent clearly showed that Pgm6/7 mainly form stable heterotrimers via intermolecular disulfide bonds. Furthermore, growth retardation and arrest of the three mutants and increased permeability of their outer membranes indicated that Pgm6/7 play an important role in outer membrane integrity. Based on results of liposome swelling experiments, these proteins are likely to function as a stabilizer of the cell wall rather than as a major porin in this organism. PMID:15659668

Nagano, Keiji; Read, Erik K; Murakami, Yukitaka; Masuda, Takashi; Noguchi, Toshihide; Yoshimura, Fuminobu

2005-02-01

133

Trimeric structure of major outer membrane proteins homologous to OmpA in Porphyromonas gingivalis.  

UK PubMed Central (United Kingdom)

The major outer membrane proteins Pgm6 (41 kDa) and Pgm7 (40 kDa) of Porphyromonas gingivalis ATCC 33277 are encoded by open reading frames pg0695 and pg0694, respectively, which form a single operon. Pgm6 and Pgm7 (Pgm6/7) have a high degree of similarity to Escherichia coli OmpA in the C-terminal region and are predicted to form eight-stranded beta-barrels in the N-terminal region. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Pgm6/7 appear as bands with apparent molecular masses of 40 and 120 kDa, with and without a reducing agent, suggesting a monomer and trimer, respectively. To verify the predicted trimeric structure and function of Pgm6/7, we constructed three mutants with pg0695, pg0694, or both deleted. The double mutant produced no Pgm6/7. The single-deletion mutants appeared to contain less Pgm7 and Pgm6 and to form homotrimers that migrated slightly faster (115 kDa) and slower (130 kDa), respectively, than wild-type Pgm6/7 under nonreducing conditions. N-terminal amino acid sequencing and mass spectrometry analysis of partially digested Pgm6/7 detected only fragments from Pgm6 and Pgm7. Two-dimensional, diagonal electrophoresis and chemical cross-linking experiments with or without a reducing agent clearly showed that Pgm6/7 mainly form stable heterotrimers via intermolecular disulfide bonds. Furthermore, growth retardation and arrest of the three mutants and increased permeability of their outer membranes indicated that Pgm6/7 play an important role in outer membrane integrity. Based on results of liposome swelling experiments, these proteins are likely to function as a stabilizer of the cell wall rather than as a major porin in this organism.

Nagano K; Read EK; Murakami Y; Masuda T; Noguchi T; Yoshimura F

2005-02-01

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Crystallization and preliminary X-ray diffraction analysis of gingipain R2 from Porphyromonas gingivalis in complex with H-D-Phe-Phe-Arg-chloromethylketone.  

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Gingipain R2 is a 50 kDa proteinase from the oral pathogenic bacterium Porphyromonas gingivalis. This proteinase, which displays no significant sequence homology to any protein previously analyzed by X-ray crystallography, has been crystallized using the vapor diffusion method. Two different crystal...

Banbula, A.; Potempa, J.; Travis, J.; Bode, W.; Medrano, F. J.

135

Experimental periodontitis induced by Porphyromonas gingivalis does not alter the onset or severity of diabetes in mice.  

UK PubMed Central (United Kingdom)

BACKGROUND AND OBJECTIVE: Diabetes mellitus is believed to increase the risk and severity of periodontitis. However, less evidence is available on the converse effects of periodontitis on diabetes. The objective of the study was to investigate to what degree experimental periodontitis induced by Porphyromonas gingivalis might influence the onset and severity of diabetes in different mouse models. MATERIAL AND METHODS: Twenty-eight male Tallyho/JngJ mice (type 2 diabetes), 20 male streptozotocin-induced diabetes C57BL/6J mice (type 1 diabetes) and 20 male C57BL/6J mice at 4 wks of age were evenly divided into two groups: periodontal infection and sham infection. Periodontitis was induced by Porphyromonas gingivalis W50 (P. gingivalis) oral inoculation before the development of diabetes. Sham-infected mice received vehicle as control. P. gingivalis in the oral cavity were identified by quantitative polymerase chain reaction. Fasting glucose, body weight and food intake levels were monitored and glucose tolerance tests were performed to assess glucose homeostasis for the onset and progression of diabetes. The level of alveolar bone loss and tumor necrosis factor-alpha were determined in week 20 when mice were killed. RESULTS: Mice in the infection groups developed more alveolar bone loss than those in sham-infection groups (Tallyho p = 0.021; C57-STZ p = 0.014; C57 p = 0.035). Hyperglycemic mice exhibited significantly more bone loss compared to those normal glucose mice (Tallyho vs. C57 p = 0.029; C57-STZ vs. C57 p = 0.024). The level of tumor necrosis factor-alpha was consistent with that of periodontal bone loss and hyperglycemia. There was no significant effect of mouse species on the amount of bone loss at the same level of blood glucose. No statistically significant difference or trend in glucose metabolism was found between the infection and sham-infection group. CONCLUSION: Diabetes enhanced the risk for periodontal disease induced by P. gingivalis. However, no converse impact was found between this periodontal infection and onset and severity of diabetes in both type 1 and 2 diabetes mice.

Li H; Yang H; Ding Y; Aprecio R; Zhang W; Wang Q; Li Y

2013-10-01

136

PURIFICACIÓN DE LIPOPOLISACÁRIDO DE Porphyromonas gingivalis LIBRE DE POLISACÁRIDOS UTILIZANDO CROMATOGRAFÍA DE ALTA RESOLUCIÓN SEPHACRYL S-200  

Directory of Open Access Journals (Sweden)

Full Text Available El objetivo de este trabajo fue mejorar un método estándar para la purificación de lipopolisacárido (LPS) de Porphyromonas gingivalis libre de polisacáridos usando una estrategia de extracción, digestión enzimática y cromatografía de alta resolución. La bacteria P. gingivalis se cultivó en condiciones de anaerobiosis y se hizo extracción de las membranas con el método de fenol-agua. Luego de una digestión enzimática (DNAsa, RNAsa y proteasa) se separó el extracto por filtración por gel con Sephacryl S-200. La muestra purificada se caracterizó por electroforesis en gel de acrilamida con tinción de plata y por el método Purpald se detecto el ácido 2-ceto-3-desoxioctulosónico (KDO). Se obtuvo una preparación libre de ácidos nucleicos, proteínas y polisacáridos. La separación por cromatografía fue de alta resolución al permitir la obtención de dos picos con diferentes componentes. El protocolo de purificación nos permitió obtener LPS de P. gingivalis con alto grado de pureza, el cual podría ser usado en próximos ensayos para evaluar su función en ensayos in vitro e in vivo; así como iniciar la obtención de LPS de otras bacterias períodontopáticas, con el fin de investigar la asociación de enfermedad períodontal con enfermedades cardiovasculares.

Gualtero Diego; Castellanos Jaime; Perez Gerardo; Lafaurie Gloria

2008-01-01

137

Porphyromonas gingivalis HmuY-induced production of interleukin-6 and IL-6 polymorphism in chronic periodontitis.  

UK PubMed Central (United Kingdom)

BACKGROUND: In chronic periodontitis (CP), the gene polymorphism of interleukin-6 (IL-6) to 174C/G has been associated with the altered production of this cytokine. The aim of this pilot study is to compare the allelic and genotypic frequencies in patients with CP with control individuals without periodontitis (NP) and to measure the production of IL-6 by whole blood cells stimulated with Porphyromonas gingivalis HmuY protein. METHODS: DNA was isolated from peripheral blood cells of 49 patients with CP and 60 control individuals classified as NP, and genotyping was performed by polymerase chain reaction using sequence-specific primers. Whole blood cells from 29 patients with CP and 30 control individuals were stimulated for 48 hours with HmuY, and IL-6 levels were measured using enzyme-linked immunosorbent assay. RESULTS: The proportion of individuals carrying the G allele at position -174 of the IL-6 gene was higher in the group with CP (85.7%) than in the normal control group (73.3%; P <0.03). P. gingivalis HmuY-induced production of IL-6 was higher in the group with CP (P <0.05). CONCLUSIONS: Our findings suggest that P. gingivalis HmuY may be associated with increased IL-6 production during CP. Furthermore, patients with periodontitis and individuals with higher HmuY-induced production of IL-6 show a high frequency of the G allele at position -174.

Trindade SC; Olczak T; Gomes-Filho IS; de Moura-Costa LF; Vale VC; Galdino-Neto M; Alves Dos Santos H; de Carvalho Filho PC; Stocker A; Bendicho MT; Xavier MT; de Moraes Marcílio Cerqueira E; Meyer R

2013-05-01

138

The inhibitory effect of alendronate and taurine on osteoclast differentiation mediated by Porphyromonas gingivalis sonicates in vitro.  

UK PubMed Central (United Kingdom)

The objective of this study was to investigate the ability of alendronate and taurine in inhibiting in vitro osteoclast differentiation induced by bacteria. Whole cell sonicates of Porphyromonas gingivalis were used as an osteoclast-stimulating factor in a mouse coculture system and differentiated osteoclasts were confirmed by tartrate-resistant acid phosphatase (TRAP) staining. Alendronate at the concentrations of 10(-7) M, 10(-6) M, and 10(-5) M and taurine at the concentrations of 4 mM, 8 mM, and 12 mM were used. The cytotoxic effects of alendronate and taurine were examined using methylthiazole-tetrazolium bromide (MTT) assay. The amounts of interleukin-6 (IL-6) in culture supernatants were also measured using ELISA. The sonicates of P. gingivalis at the concentration of 0.01-0.1 microg/ml significantly stimulated the formation of osteoclasts (p < 0.05). Alendronate (10(-5) M) and taurine (12 mM) significantly suppressed the sonicate-stimulated osteoclast formation. In MTT assay, no cytotoxic effects were evident in all concentrations of alendronate and taurine. Alendronate and taurine did not affect the amount of IL-6 induced by P. gingivalis sonicates. These data indicate that alendronate and taurine have inhibitory effects on bacteria-stimulated osteoclast formation in vitro and that this inhibitory mechanism is not related to the blocking of IL-6 production.

Kum KY; Park JH; Yoo YJ; Choi BK; Lee HJ; Lee SJ

2003-01-01

139

Clinical correlations with Porphyromonas gingivalis antibody responses in patients with early rheumatoid arthritis.  

UK PubMed Central (United Kingdom)

INTRODUCTION: Prior studies have demonstrated increased frequencies of antibodies to Porphyromonas gingivalis (Pg), a leading agent of periodontal disease (PD), in rheumatoid arthritis (RA) patients. However, these patients generally had longstanding disease, and clinical associations with these antibodies were inconsistent. Our goal was to examine Pg antibody responses and their clinical associations in patients with early RA, prior to and after disease-modifying antirheumatic drug (DMARD) therapy. METHODS: Serum samples were tested from 50 DMARD-naive RA patients using an ELISA with whole Pg sonicate. For comparison, serum samples were tested from patients with late RA, other connective tissue diseases (CTD), age-similar healthy hospital personnel, and blood bank donors. Pg antibody responses in early RA patients were correlated with standard RA biomarkers, measures of disease activity, and function. RESULTS: At enrollment, 17 of the 50 patients (34%) with early RA had positive IgG antibody responses to Pg, as did 13 of the 43 patients (30%) with late RA. RA patients had significantly higher Pg antibody responses than healthy hospital personnel and blood bank donors (P<0.0001). Additionally, RA patients tended to have higher Pg antibody reactivity than patients with other CTD (P=0.1), and CTD patients tended to have higher Pg responses than healthy subjects (P=0.07). Compared with Pg antibody-negative patients, early RA patients with positive Pg responses more often had anti-CCP antibody reactivity, their anti-CCP levels were significantly higher (P=0.03), and the levels of anti-Pg antibodies correlated directly with anti-CCP levels (P<0.01). Furthermore, at study entry, the Pg-positive antibody group had greater RF values (P=0.04) and higher inflammatory markers (erythrocyte sedimentation rate, ESR) (P=0.05), and tended to have higher disease activity scores (DAS-28-ESR and CDAI), and more functional impairment (HAQ). In Pg-positive patients, greater disease activity was still apparent after 12 months of DMARD therapy. CONCLUSIONS: A subset of early RA patients had positive Pg antibody responses; the responses correlated with anti-CCP antibody reactivity, and to a lesser degree with ESR values. There was a trend toward greater disease activity in Pg-positive patients, and this trend remained after 12 months of DMARD therapy. These findings are consistent with a role for Pg in disease pathogenesis in a subset of RA patients.

Arvikar SL; Collier DS; Fisher MC; Unizony S; Cohen GL; McHugh G; Kawai T; Strle K; Steere AC

2013-09-01

140

In vitro cytokine responses to periodontal pathogens: generalized aggressive periodontitis is associated with increased IL-6 response to Porphyromonas gingivalis  

DEFF Research Database (Denmark)

Generalized aggressive periodontitis (GAgP) is an inflammatory condition resulting in destruction of tooth-supporting tissues. We examined the production of IL-1beta, IL-6, tumour necrosis factor (TNF)-alpha, IL-12 and IL-10 in cultures of peripheral mononuclear cells (MNC) from 10 patients with GAgP and 10 controls stimulated with periodontal pathogens or a control antigen, tetanus toxoid (TT) in the presence of autologous serum. The pathogens used were Porphyromonas gingivalis, Prevotella intermedia and Fusobacterium nucleatum, either as type strains or bacteria isolated from the participants' inherent oral flora. The P. gingivalis -induced production of IL-6 was approximately 2.5-fold higher in patients with GAgP than in healthy controls (P <0.05), while the corresponding TNF-alpha production was non-significantly elevated. IL-1beta production induced by P. gingivalis, as all cytokine responses induced by Pr. intermedia, F. nucleatum and TT was similar in the two groups. A reduced IL-12p70 response to Pr. intermedia and F. nucleatum was observed in smokers compared to non-smoking patients (P <0.02). To assess the role of serum factors in the elevated IL-6 response to P. gingivalis, MNC from two donors free of disease were stimulated with this bacterium in the presence of the various patient and control sera. An elevated IL-6 and TNF-alpha response was observed in the presence of patient sera (P <0.01 and P <0.04, respectively). The data suggest that an exaggerated production of IL-6 occurs in GAgP, and that pro-inflammatory serum factors play an essential role in the response.

Borch, T S; Holmstrup, Palle

2010-01-01

 
 
 
 
141

Analysis of viable vs. dead Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis using selective quantitative real-time PCR with propidium monoazide.  

UK PubMed Central (United Kingdom)

BACKGROUND AND OBJECTIVES: One of the major disadvantages of DNA-based microbial diagnostics is their inability to differentiate DNA between viable and dead microorganisms, which could be important when studying etiologically relevant pathogens. The aim of this investigation was to optimize a method for the selective detection and quantification of only viable Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis cells by combining quantitative real-time polymerase chain reaction (qPCR) and propidium monoazide (PMA). MATERIAL AND METHODS: Three different concentrations of PMA (10, 50 or 100 ?m) were added to suspensions of 10(6) (CFU)/mL of viable/dead A. actinomycetemcomitans and P. gingivalis cells. After DNA isolation, qPCR was carried out using specific primers and probes for the tested bacteria. PMA was further tested with different mixtures containing varying ratios of viable and dead cells. The efficacy of PMA to detect viable/dead cells was tested by analysis of variance. RESULTS: For these specific bacterial pathogens, 100 ?m PMA resulted in a significant reduction of qPCR amplification with dead cells (10(6) CFU/mL), while with viable cells no significant inhibition was detected. PMA was also effective in detecting selectively viable cells by qPCR detection, when mixtures of varying ratios of viable and dead bacteria were used. CONCLUSIONS: This study demonstrated the efficiency of PMA for differentiating viable and dead A. actinomycetemcomitans and P. gingivalis cells. This method of PMA-qPCR may be useful for monitoring new antimicrobial strategies and for assessing the pathogenic potential of A. actinomycetemcomitans and P. gingivalis in different oral conditions when using molecular diagnostic methods.

Sánchez MC; Marín MJ; Figuero E; Llama-Palacios A; Herrera D; Sanz M

2013-04-01

142

Immunogenicity of a cholera toxin B subunit Porphyromonas gingivalis fimbrial antigen fusion protein expressed in E. coli.  

Science.gov (United States)

The gram-negative anaerobic oral bacterium Porphyromonas gingivalis initiates periodontal disease through fimbrial attachment to saliva-coated oral surfaces. To study the effects of immunomodulation on enhancement of subunit vaccination, the expression in E. coli and immunogenicity of P. gingivalis fimbrial protein (FimA) linked to the C-terminus of the cholera toxin B subunit (CTB) were investigated. Complementary DNAs encoding the P. gingivalis 381 fimbrillin protein sequence FimA1 (amino acid residues 1-200) and FimA2 (amino acid residues 201-337) were cloned into an E. coli expression vector downstream of a cDNA fragment encoding the immunostimulatory CTB. CTB-FimA1 and CTB-FimA2 fusion proteins synthesized in E. coli BL21 (DE3) cells were purified under denaturing conditions by Ni2+-NTA affinity column chromatography. Renaturation of the CTB-FimA1 and CTB-FimA2 fusion proteins, permitted identification of CTB-FimA pentamers and restored CTB binding activity to GM1-ganglioside to provide a biologically active CTB-FimA fusion protein. Mice orally inoculated with purified CTB-FimA1 or CTB-FimA2 fusion proteins generated measurable FimA1 and FimA2 IgG antibody titers, while no serum fimbrial IgG antibodies were detected when mice were inoculated with FimA1 or FimA2 proteins alone. Immunoblot analysis confirmed that sera from mice immunized with CTB linked to FimA1 or FimA2 contained antibodies specific for P. gingivalis fimbrial proteins. In addition, mice immunized with FimA2 or CTB-FimA2 generated measurable intestinal IgA titers indicating the presence of fimbrial antibody class switching. Further, mice orally immunized with CTB-FimA1 generated higher IgA antibody titers than mice inoculated with FimA1 alone. The experimental data show that the immunostimulatory molecule CTB enhances B cell-mediated immunity against linked P. gingivalis FimA fusion proteins, in comparison to immunization with FimA protein alone. Thus, linkage of CTB to P. gingivalis fimbrial antigens can increase subunit vaccine immunogenicity to provide enhanced protection against periodontal disease. PMID:18807220

Kim, Tae-Geum; Huy, Nguyen-Xuan; Kim, Mi-Young; Jeong, Dong-Keun; Jang, Yong-Suk; Yang, Moon-Sik; Langridge, William H R; Lee, Jin-Yong

2008-09-20

143

Immunogenicity of a cholera toxin B subunit Porphyromonas gingivalis fimbrial antigen fusion protein expressed in E. coli.  

UK PubMed Central (United Kingdom)

The gram-negative anaerobic oral bacterium Porphyromonas gingivalis initiates periodontal disease through fimbrial attachment to saliva-coated oral surfaces. To study the effects of immunomodulation on enhancement of subunit vaccination, the expression in E. coli and immunogenicity of P. gingivalis fimbrial protein (FimA) linked to the C-terminus of the cholera toxin B subunit (CTB) were investigated. Complementary DNAs encoding the P. gingivalis 381 fimbrillin protein sequence FimA1 (amino acid residues 1-200) and FimA2 (amino acid residues 201-337) were cloned into an E. coli expression vector downstream of a cDNA fragment encoding the immunostimulatory CTB. CTB-FimA1 and CTB-FimA2 fusion proteins synthesized in E. coli BL21 (DE3) cells were purified under denaturing conditions by Ni2+-NTA affinity column chromatography. Renaturation of the CTB-FimA1 and CTB-FimA2 fusion proteins, permitted identification of CTB-FimA pentamers and restored CTB binding activity to GM1-ganglioside to provide a biologically active CTB-FimA fusion protein. Mice orally inoculated with purified CTB-FimA1 or CTB-FimA2 fusion proteins generated measurable FimA1 and FimA2 IgG antibody titers, while no serum fimbrial IgG antibodies were detected when mice were inoculated with FimA1 or FimA2 proteins alone. Immunoblot analysis confirmed that sera from mice immunized with CTB linked to FimA1 or FimA2 contained antibodies specific for P. gingivalis fimbrial proteins. In addition, mice immunized with FimA2 or CTB-FimA2 generated measurable intestinal IgA titers indicating the presence of fimbrial antibody class switching. Further, mice orally immunized with CTB-FimA1 generated higher IgA antibody titers than mice inoculated with FimA1 alone. The experimental data show that the immunostimulatory molecule CTB enhances B cell-mediated immunity against linked P. gingivalis FimA fusion proteins, in comparison to immunization with FimA protein alone. Thus, linkage of CTB to P. gingivalis fimbrial antigens can increase subunit vaccine immunogenicity to provide enhanced protection against periodontal disease.

Kim TG; Huy NX; Kim MY; Jeong DK; Jang YS; Yang MS; Langridge WH; Lee JY

2009-02-01

144

Prevotella intermedia and Porphyromonas gingivalis isolated from osseointegrated dental implants: colonization and antimicrobial susceptibility/ Prevotella intermedia e Porphyromonas gingivalis isolados de implantes osseointegrados: colonização e susceptibilidade a antimicrobianos  

Scientific Electronic Library Online (English)

Full Text Available Abstract in portuguese Neste estudo foram avaliadas a colonização e a susceptibilidade a antimicrobianos de P. intermedia e P. gingivalis isolados de amostras de sulcus gengivais e peri-implantares. As amostras foram coletadas de 30 pacientes submetidos a implantes, em três tempos diferentes: no momento da cirurgia, 20 e 60 dias após a instalação do implante. Os organismos foram identificados por testes bioquímicos ou por kit comercial API 32-A e por PCR. A susceptibilidade antimicrobian (more) a foi determinada usando-se o método de diluição em ágar. Foram isolados dezenove P. intermedia (quatro de peri-implantites e 15 de sulco gengival) e somente sete P. gingivalis de sulco gengival. Pelo PCR os organismos foram detectados de sete amostras sete peri-implantares e de 32 gengivais. As bactérias foram susceptíveis aos antibióticos usados exceto para azitromicina com 65% de resistência para P. intermedia. As espécies avaliadas foram sensíveis para cádmio, níquel e paládio, e mostraram diferentes faixas de resistência para titânio, alumínio e bicloreto de mercúrio. A maioria de P. intermedia foi resistente para chumbo, prata, cobre, titânio, zinco, alumínio e bicloreto de mercúrio. As bactérias colonizaram implantes após 60 dias de cirurgia e PCR pode ser usado como ferramenta para a detecção bacteriana na implantodontia. Abstract in english The colonization and antimicrobial susceptibility of P. intermedia and P. gingivalis isolated from peri-implant and gingival sulcus samples were determined. Samples were collected from 30 patients submitted to implant in three different times: at the moment of the surgery, 20 and 60 days after the implant installation. Organisms were identified by using a biochemical tests or API 32-A kit and by PCR. The antimicrobial susceptibility was determined by using an agar dilutio (more) n method. Nineteen P. intermedia (4 from peri-implant sites and 15 from gingival sulcus), and only seven P. gingivalis from gingival sulcus were isolated. Organisms were detected by PCR from seven peri-implant and 32 gingival samples. Bacteria were susceptible to the used antibiotics except to azithromycin with 65% of resistance for P. intermedia strains. Both tested species were susceptible to cadmium, nickel and palladium, and showed different resistance rates to titanium, aluminum and mercuric chloride. Most of P. intermedia strains were resistant to lead, silver, copper, titanium, zinc, aluminum and mercuric chloride. Bacteria colonized implants after 60 days of surgery and PCR may be used as a tool for bacterial detection in implantology.

Pfau, Eduardo Augusto; Avila-Campos, Mario Julio

2005-09-01

145

Prevotella intermedia and Porphyromonas gingivalis isolated from osseointegrated dental implants: colonization and antimicrobial susceptibility Prevotella intermedia e Porphyromonas gingivalis isolados de implantes osseointegrados: colonização e susceptibilidade a antimicrobianos  

Directory of Open Access Journals (Sweden)

Full Text Available The colonization and antimicrobial susceptibility of P. intermedia and P. gingivalis isolated from peri-implant and gingival sulcus samples were determined. Samples were collected from 30 patients submitted to implant in three different times: at the moment of the surgery, 20 and 60 days after the implant installation. Organisms were identified by using a biochemical tests or API 32-A kit and by PCR. The antimicrobial susceptibility was determined by using an agar dilution method. Nineteen P. intermedia (4 from peri-implant sites and 15 from gingival sulcus), and only seven P. gingivalis from gingival sulcus were isolated. Organisms were detected by PCR from seven peri-implant and 32 gingival samples. Bacteria were susceptible to the used antibiotics except to azithromycin with 65% of resistance for P. intermedia strains. Both tested species were susceptible to cadmium, nickel and palladium, and showed different resistance rates to titanium, aluminum and mercuric chloride. Most of P. intermedia strains were resistant to lead, silver, copper, titanium, zinc, aluminum and mercuric chloride. Bacteria colonized implants after 60 days of surgery and PCR may be used as a tool for bacterial detection in implantology.Neste estudo foram avaliadas a colonização e a susceptibilidade a antimicrobianos de P. intermedia e P. gingivalis isolados de amostras de sulcus gengivais e peri-implantares. As amostras foram coletadas de 30 pacientes submetidos a implantes, em três tempos diferentes: no momento da cirurgia, 20 e 60 dias após a instalação do implante. Os organismos foram identificados por testes bioquímicos ou por kit comercial API 32-A e por PCR. A susceptibilidade antimicrobiana foi determinada usando-se o método de diluição em ágar. Foram isolados dezenove P. intermedia (quatro de peri-implantites e 15 de sulco gengival) e somente sete P. gingivalis de sulco gengival. Pelo PCR os organismos foram detectados de sete amostras sete peri-implantares e de 32 gengivais. As bactérias foram susceptíveis aos antibióticos usados exceto para azitromicina com 65% de resistência para P. intermedia. As espécies avaliadas foram sensíveis para cádmio, níquel e paládio, e mostraram diferentes faixas de resistência para titânio, alumínio e bicloreto de mercúrio. A maioria de P. intermedia foi resistente para chumbo, prata, cobre, titânio, zinco, alumínio e bicloreto de mercúrio. As bactérias colonizaram implantes após 60 dias de cirurgia e PCR pode ser usado como ferramenta para a detecção bacteriana na implantodontia.

Eduardo Augusto Pfau; Mario Julio Avila-Campos

2005-01-01

146

Further evidence that major outer membrane proteins homologous to OmpA in Porphyromonas gingivalis stabilize bacterial cells.  

UK PubMed Central (United Kingdom)

INTRODUCTION: Porphyromonas gingivalis is one of the most important bacteria in the progression of chronic periodontal disease. We hypothesized that the major outer membrane proteins Pgm6/7, which are homologous to the OmpA protein in Escherichia coli, might contribute to the stabilization of the cell surface. In this study, the effects of Pgm6/7 on the cell surface were examined morphologically. METHODS: Deletion mutants of Pgm6/7 (Delta694, Delta695 and Delta695-694) were constructed using the polymerase chain reaction-based overlap extension method. Wild-type ATCC 33277 and Pgm6/7 mutants were grown under anaerobic conditions. Whole cells and thin sections of fixed cells were stained and examined by transmission electron microscopy. RESULTS: Compared with the wild-type, numerous vesicles released from cells were observed in each deletion mutant. The outer membrane appeared wavy and irregular. Increased numbers of vesicles were confirmed after their preparation from the culture supernatant. Total gingipain activity in vesicles was increased five- to 10-fold in the deletion mutants. CONCLUSION: This report provides further evidence that Pgm6/7 proteins in P. gingivalis play an important role in the maintenance of bacterial outer membrane integrity.

Iwami J; Murakami Y; Nagano K; Nakamura H; Yoshimura F

2007-10-01

147

Sublingual vaccination with outer membrane protein of Porphyromonas gingivalis and Flt3 ligand elicits protective immunity in the oral cavity.  

Science.gov (United States)

In this study, we demonstrated that the 40-kDa outer membrane protein of Porphyromonas gingivalis (40k-OMP) sublingually administered with a cDNA vector plasmid encoding Flt3 ligand (pFL) elicited a protective immune response. Sublingual immunization of mice with 40k-OMP plus pFL induced significant serum IgG and IgA, as well as salivary IgA, antibody responses that were comparable to those induced by 40k-OMP plus cholera toxin as adjuvant. When the subclasses of 40k-OMP-specific IgG were evaluated, sublingual immunization with 40k-OMP plus pFL induced both IgG1 and IgG2a antibody responses. Sublingual delivery of pFL resulted in FL expression in submandibular glands, but not in other oral tissues. Furthermore, marked increases in FL protein occurred in saliva and serum, and the frequencies of both CD11c(+)CD11b(+) and CD11c(+)CD8alpha(+) dendritic cells with up-regulated expression of CD80, CD86 and CD40 molecules significantly increased in submandibular lymph nodes and spleen. Importantly, the mice given sublingual 40k-OMP plus pFL showed a significant reduction of alveolar bone loss caused by oral infection with P. gingivalis. These findings suggest that sublingual administration of 40k-OMP with pFL acts as an effective and safe mucosal vaccine against oral P. gingivalis infection, and may be a useful tool in the prevention of chronic periodontitis. PMID:19852927

Zhang, Tao; Hashizume, Tomomi; Kurita-Ochiai, Tomoko; Yamamoto, Masafumi

2009-10-21

148

Sublingual vaccination with outer membrane protein of Porphyromonas gingivalis and Flt3 ligand elicits protective immunity in the oral cavity.  

UK PubMed Central (United Kingdom)

In this study, we demonstrated that the 40-kDa outer membrane protein of Porphyromonas gingivalis (40k-OMP) sublingually administered with a cDNA vector plasmid encoding Flt3 ligand (pFL) elicited a protective immune response. Sublingual immunization of mice with 40k-OMP plus pFL induced significant serum IgG and IgA, as well as salivary IgA, antibody responses that were comparable to those induced by 40k-OMP plus cholera toxin as adjuvant. When the subclasses of 40k-OMP-specific IgG were evaluated, sublingual immunization with 40k-OMP plus pFL induced both IgG1 and IgG2a antibody responses. Sublingual delivery of pFL resulted in FL expression in submandibular glands, but not in other oral tissues. Furthermore, marked increases in FL protein occurred in saliva and serum, and the frequencies of both CD11c(+)CD11b(+) and CD11c(+)CD8alpha(+) dendritic cells with up-regulated expression of CD80, CD86 and CD40 molecules significantly increased in submandibular lymph nodes and spleen. Importantly, the mice given sublingual 40k-OMP plus pFL showed a significant reduction of alveolar bone loss caused by oral infection with P. gingivalis. These findings suggest that sublingual administration of 40k-OMP with pFL acts as an effective and safe mucosal vaccine against oral P. gingivalis infection, and may be a useful tool in the prevention of chronic periodontitis.

Zhang T; Hashizume T; Kurita-Ochiai T; Yamamoto M

2009-12-01

149

[Relationship between volatile fatty acids and Porphyromonas gingivalis and Treponema denticola in gingival crevicular fluids of patients with aggressive periodontitis].  

UK PubMed Central (United Kingdom)

OBJECTIVE: Short chain fatty acids (SCFAs), such as succinic acid, acetic acid, propionic acid, butyric acid, etc. are metabolic product of putative periodontal pathogens, which play significant roles in periodontitis. The aim of this study was to analyze the relationship between Porphyromonas gingivalis (P. gingivalis), Treponema denticola (T. denticola), and the concentration of SCFAs in gingival crevicular fluid (GCF) of patients with aggressive periodontitis (AgP). METHODS: GCF was sampled from 4 sites per individual in 20 patients with AgP and 14 healthy controls. Concentrations of SCFAs, including succinic acid, acetic acid, propionic acid, butyric acid, and isovaleric acid in the supernant of GCF were analyzed by high performance capillary electrophoresis (HPCE), P. gingivalis and T. denticola in the deposit of the same GCF were detected by PCR with their electrophoretic band quantified. RESULTS: The concentrations of succinic acid, acetic acid, propionic acid, butyric acid, and isovaleric acid, the prevalence and PCR band quantity of P. gingivalis and T. denticola in GCF were all significantly higher in patients with AgP than that of healthy controls. In patients with AgP, butyric acid concentration was significantly higher in P. gingivalis positive sites than negative sites [2.87 (0.99, 4.36) mmol/L vs. 0.33 (0.00, 1.44) mmol/L, P<0.05], the concentrations of succinic acid, acetic acid, propionic acid, butyric acid, and isovaleric acid were positively correlated with PCR band quantity of P. gingivalis (r value was 0.334, 0.548, 0.411, 0.493, 0.273, respectively, P<0.05); the concentrations of SCFAs were significantly higher in T. denticola positive sites than negative sites: succinic acid, 1.67 (1.15, 2.11) mmol/L vs. 0.80 (0.48, 1.06) mmol/L; acetic acid, 31.95 (23.77, 43.13) mmol/L vs.12.51 (7.57, 15.69) mmol/L; propionic acid, 11.86 (6.55, 14.98) mmol/L vs. 2.82 (1.71, 7.03) mmol/L; butyric acid, 3.45 (2.41, 4.78) mmol/L vs. 0.54 (0.00, 1.56) mmol/L; isovaleric acid, 2.23 (1.05, 3.85) mmol/L vs. 0.62 (0.00, 2.33) mmol/L. The concentrations of succinic acid, acetic acid, propionic acid, butyric acid were positively correlated with PCR band quantity of T. denticola (r value was 0.443, 0.702, 0.625, 0.557, respectively, P<0.05). CONCLUSION: SCFAs concentrations reflect the quantity of P. gingivalis and T. denticola in patients with AgP, and may be an indicator to the disease progression in patients with AgP.

Lu RF; Feng L; Gao XJ; Meng HX; Feng XH

2013-02-01

150

Emerging family of proline-specific peptidases of Porphyromonas gingivalis: purification and characterization of serine dipeptidyl peptidase, a structural and functional homologue of mammalian prolyl dipeptidyl peptidase IV.  

Science.gov (United States)

Porphyromonas gingivalis is an asaccharolytic and anaerobic bacterium that possesses a complex proteolytic system which is essential for its growth and evasion of host defense mechanisms. In this report, we show the purification and characterization of prolyl dipeptidyl peptidase IV (DPPIV) produced by this organism. The enzyme was purified to homogeneity, and its enzymatic activity and biochemical properties were investigated. P. gingivalis DPPIV, like its human counterpart, is able to cleave the N terminus of synthetic oligopeptides with sequences analogous to those of interleukins 1beta and 2. Additionally, this protease hydrolyzes biologically active peptides including substance P, fibrin inhibitory peptide, and beta-casomorphin. Southern blot analysis of genomic DNA isolated from several P. gingivalis strains reveal that a single copy of the DPPIV gene was present in all strains tested. PMID:10678923

Banbula, A; Bugno, M; Goldstein, J; Yen, J; Nelson, D; Travis, J; Potempa, J

2000-03-01

151

Arg-gingipain inhibition and anti-bacterial activity selective for Porphyromonas gingivalis by malabaricone C.  

UK PubMed Central (United Kingdom)

Effects of malabaricon C, isolated from nutmeg (Myristica fragrans), on Arg-gingipain activity and growth of several kinds of anaerobic and aerobic microorganisms were investigated. Malabaricone C irreversibly inhibited Arg-gingipain by 50% at a concentration of 0.7 microgram/ml and selectively suppressed Porphyromomas gingivalis growth.

Shinohara C; Mori S; Ando T; Tsuji T

1999-08-01

152

Alkyl hydroperoxide peroxidase subunit C (ahpC) protects against organic peroxides but does not affect the virulence of Porphyromonas gingivalis W83.  

UK PubMed Central (United Kingdom)

The cloned Porphyromonas gingivalis alkyl hydroperoxide reductase (ahpC) gene complemented an ahpC defect in Escherichia coli. To study the role of ahpC in protecting against oxidative stress in P. gingivalis a 1.8 kb fragment containing the ahpC gene was amplified from the chromosome of P. gingivalis W83. This gene was insertionally inactivated using the ermF-ermAM antibiotic resistance cassette and used to create a ahpC-deficient mutant by allelic exchange. One mutant strain, designated FLL141, demonstrated no change in the growth rate, black pigmentation, beta-hemolysis or level of proteolytic activity compared to the parent strain. Although P. gingivalis FLL141 was more sensitive to hydrogen peroxide than the parent strain, there was no change in its virulence potential in the mouse model compared to the wild-type strain. These findings suggest that the ahpC gene plays a role in peroxide resistance in P. gingivalis but does not contribute significantly to virulence.

Johnson NA; Liu Y; Fletcher HM

2004-08-01

153

Alkyl hydroperoxide peroxidase subunit C (ahpC) protects against organic peroxides but does not affect the virulence of Porphyromonas gingivalis W83.  

Science.gov (United States)

The cloned Porphyromonas gingivalis alkyl hydroperoxide reductase (ahpC) gene complemented an ahpC defect in Escherichia coli. To study the role of ahpC in protecting against oxidative stress in P. gingivalis a 1.8 kb fragment containing the ahpC gene was amplified from the chromosome of P. gingivalis W83. This gene was insertionally inactivated using the ermF-ermAM antibiotic resistance cassette and used to create a ahpC-deficient mutant by allelic exchange. One mutant strain, designated FLL141, demonstrated no change in the growth rate, black pigmentation, beta-hemolysis or level of proteolytic activity compared to the parent strain. Although P. gingivalis FLL141 was more sensitive to hydrogen peroxide than the parent strain, there was no change in its virulence potential in the mouse model compared to the wild-type strain. These findings suggest that the ahpC gene plays a role in peroxide resistance in P. gingivalis but does not contribute significantly to virulence. PMID:15209993

Johnson, N A; Liu, Y; Fletcher, H M

2004-08-01

154

Inactivation of vimF, a putative glycosyltransferase gene downstream of vimE, alters glycosylation and activation of the gingipains in Porphyromonas gingivalis W83.  

UK PubMed Central (United Kingdom)

Regulation/activation of the Porphyromonas gingivalis gingipains is poorly understood. A 1.2-kb open reading frame, a putative glycosyltransferase, downstream of vimE, was cloned, insertionally inactivated using the ermF-ermAM antibiotic resistance cassette, and used to create a defective mutant by allelic exchange. In contrast to the wild-type W83 strain, this mutant, designated P. gingivalis FLL95, was nonpigmented and nonhemolytic when plated on Brucella blood agar. Arginine- and lysine-specific gingipain activities were reduced by approximately 97% and 96%, respectively, relative to that of the parent strain. These activities were unaffected by the growth phase, in contrast to the vimA-defective mutant P. gingivalis FLL92. Expression of the rgpA, rgpB, and kgp gingipain genes was unaffected in P. gingivalis FLL95 in comparison to the wild-type strain. In nonactive gingipain extracellular protein fractions, multiple high-molecular-weight proteins immunoreacted with gingipain-specific antibodies. The specific gingipain-associated sugar moiety recognized by monoclonal antibody 1B5 was absent in FLL95. Taken together, these results suggest that the vimE downstream gene, designated vimF (virulence modulating gene F), which is a putative glycosyltransferase group 1, is involved in the regulation of the major virulence factors of P. gingivalis.

Vanterpool E; Roy F; Fletcher HM

2005-07-01

155

Inactivation of vimF, a putative glycosyltransferase gene downstream of vimE, alters glycosylation and activation of the gingipains in Porphyromonas gingivalis W83.  

Science.gov (United States)

Regulation/activation of the Porphyromonas gingivalis gingipains is poorly understood. A 1.2-kb open reading frame, a putative glycosyltransferase, downstream of vimE, was cloned, insertionally inactivated using the ermF-ermAM antibiotic resistance cassette, and used to create a defective mutant by allelic exchange. In contrast to the wild-type W83 strain, this mutant, designated P. gingivalis FLL95, was nonpigmented and nonhemolytic when plated on Brucella blood agar. Arginine- and lysine-specific gingipain activities were reduced by approximately 97% and 96%, respectively, relative to that of the parent strain. These activities were unaffected by the growth phase, in contrast to the vimA-defective mutant P. gingivalis FLL92. Expression of the rgpA, rgpB, and kgp gingipain genes was unaffected in P. gingivalis FLL95 in comparison to the wild-type strain. In nonactive gingipain extracellular protein fractions, multiple high-molecular-weight proteins immunoreacted with gingipain-specific antibodies. The specific gingipain-associated sugar moiety recognized by monoclonal antibody 1B5 was absent in FLL95. Taken together, these results suggest that the vimE downstream gene, designated vimF (virulence modulating gene F), which is a putative glycosyltransferase group 1, is involved in the regulation of the major virulence factors of P. gingivalis. PMID:15972484

Vanterpool, Elaine; Roy, Francis; Fletcher, Hansel M

2005-07-01

156

Porphyromonas gingivalis-nucleoside-diphosphate-kinase inhibits ATP-induced reactive-oxygen-species via P2X7 receptor/NADPH-oxidase signalling and contributes to persistence.  

UK PubMed Central (United Kingdom)

Ligation of P2X7 receptors with a 'danger signal', extracellular ATP (eATP), has recently been shown to result in production of intracellular reactive-oxygen-species (ROS) in macrophages. We show that primary gingival epithelial cells (GECs) produce sustained, robust cellular ROS upon stimulation by eATP. The induction of ROS was mediated by P2X7 receptor signalling coupled with NADPH-oxidase activation, as determined by pharmacological inhibition and RNA interference. Furthermore, Porphyromonas gingivalis, an oral opportunistic pathogen, upregulated the antioxidant glutathione response, modulated eATP-induced cytosolic and mitochondrial ROS generated through P2X7 /NADPH-oxidase interactome, and subsequently blocked oxidative stress in GECs via temporal secretion of a P.?gingivalis effector, nucleoside-diphosphate-kinase (Ndk). An ndk-deficient P.?gingivalis mutant lacked the ability to inhibit ROS production and persist intracellularly following eATP stimulation. Treatment with recombinant Ndk significantly diminished eATP-evoked ROS production. P.?gingivalis infection elicited a strong, time-dependent increase in anti-oxidativemitochondrial UCP2 levels, whereas ndk-deficient mutant did not cause any change. The results reveal a novel signalling cascade that is tightly coupled with eATP signalling and ROS regulation. Ndk by P.?gingivalis counteracts these antimicrobial signalling activities by secreting Ndk, thus contributing to successful persistence of the pathogen.

Choi CH; Spooner R; DeGuzman J; Koutouzis T; Ojcius DM; Yilmaz Ö

2013-06-01

157

Serine Lipids of Porphyromonas gingivalis Are Human and Mouse Toll-Like Receptor 2 Ligands.  

Science.gov (United States)

The total cellular lipids of Porphyromas gingivalis, a known periodontal pathogen, were previously shown to promote dendritic cell activation and inhibition of osteoblasts through engagement of Toll-like receptor 2 (TLR2). The purpose of the present investigation was to fractionate all lipids of P. gingivalis and define which lipid classes account for the TLR2 engagement, based on both in vitro human cell assays and in vivo studies in mice. Specific serine-containing lipids of P. gingivalis, called lipid 654 and lipid 430, were identified in specific high-performance liquid chromatography fractions as the TLR2-activating lipids. The structures of these lipids were defined using tandem mass spectrometry and nuclear magnetic resonance methods. In vitro, both lipid 654 and lipid 430 activated TLR2-expressing HEK cells, and this activation was inhibited by anti-TLR2 antibody. In contrast, TLR4-expressing HEK cells failed to be activated by either lipid 654 or lipid 430. Wild-type (WT) or TLR2-deficient (TLR2(-/-)) mice were injected with either lipid 654 or lipid 430, and the effects on serum levels of the chemokine CCL2 were measured 4 h later. Administration of either lipid 654 or lipid 430 to WT mice resulted in a significant increase in serum CCL2 levels; in contrast, the administration of lipid 654 or lipid 430 to TLR2(-/-) mice resulted in no increase in serum CCL2. These results thus identify a new class of TLR2 ligands that are produced by P. gingivalis that likely play a significant role in mediating inflammatory responses both at periodontal sites and, potentially, in other tissues where these lipids might accumulate. PMID:23836823

Clark, Robert B; Cervantes, Jorge L; Maciejewski, Mark W; Farrokhi, Vahid; Nemati, Reza; Yao, Xudong; Anstadt, Emily; Fujiwara, Mai; Wright, Kyle T; Riddle, Caroline; La Vake, Carson J; Salazar, Juan C; Finegold, Sydney; Nichols, Frank C

2013-07-08

158

A monoclonal antibody against fimA type II Porphyromonas gingivalis inhibits IL-8 production in human gingival fibroblasts.  

UK PubMed Central (United Kingdom)

The periodontal pathogen Porphyromans gingivalis is classified into six groups (types I-V and Ib) based on the genotype of the fimbriae A (fimA) gene. Among genotypes, fimA type II strains are thought to be most strongly related to advanced periodontitis. The present study was undertaken to develop passive immunotherapy monoclonal antibodies (MAbs) against periodontitis, which are capable of inhibiting virulency and were constructed through the immunization of outer membrane vesicles (OMV) fraction of fimAII strain, TDC60, using mouse hybridoma technology. MAbs that recognized OMV by ELISA assay were identified, and 28 clones were screened by Western blot analysis. After purifying these MAbs using protein G column, the effect of the MAb on IL-8 production from human gingival fibroblasts by OMV was examined. We selected MAb TDC4-33H, which strongly inhibited the IL-8 production with a higher MAb production rate. Since the MAb showed an individual ladder-like profile against OMV by Western blotting, we further examined the reactivity against lipopolysaccharides (LPS) from TDC60, W83 (fimAIV), and ATCC33277 (fimAI). As a result, MAb TDC4-33H recognized all LPSs. Moreover, MAb TDC4-33H significantly inhibited the LPS-stimulated IL-8 production in human gingival fibroblasts. These findings suggest that MAb TDC4-33H reacts with LPS and may be useful for passive immunotherapy through neutralizing IL-8 production in gingival fibroblasts by P. gingivalis LPS.

Hijiya T; Shibata Y; Hayakawa M; Abiko Y

2010-06-01

159

A monoclonal antibody against fimA type II Porphyromonas gingivalis inhibits IL-8 production in human gingival fibroblasts.  

Science.gov (United States)

The periodontal pathogen Porphyromans gingivalis is classified into six groups (types I-V and Ib) based on the genotype of the fimbriae A (fimA) gene. Among genotypes, fimA type II strains are thought to be most strongly related to advanced periodontitis. The present study was undertaken to develop passive immunotherapy monoclonal antibodies (MAbs) against periodontitis, which are capable of inhibiting virulency and were constructed through the immunization of outer membrane vesicles (OMV) fraction of fimAII strain, TDC60, using mouse hybridoma technology. MAbs that recognized OMV by ELISA assay were identified, and 28 clones were screened by Western blot analysis. After purifying these MAbs using protein G column, the effect of the MAb on IL-8 production from human gingival fibroblasts by OMV was examined. We selected MAb TDC4-33H, which strongly inhibited the IL-8 production with a higher MAb production rate. Since the MAb showed an individual ladder-like profile against OMV by Western blotting, we further examined the reactivity against lipopolysaccharides (LPS) from TDC60, W83 (fimAIV), and ATCC33277 (fimAI). As a result, MAb TDC4-33H recognized all LPSs. Moreover, MAb TDC4-33H significantly inhibited the LPS-stimulated IL-8 production in human gingival fibroblasts. These findings suggest that MAb TDC4-33H reacts with LPS and may be useful for passive immunotherapy through neutralizing IL-8 production in gingival fibroblasts by P. gingivalis LPS. PMID:20568993

Hijiya, Takahiro; Shibata, Yasuko; Hayakawa, Mitsuo; Abiko, Yoshimitsu

2010-06-01

160

[Quantitative study of porphyromonas gingivalis in subgingival plaques of orthodontic adults].  

UK PubMed Central (United Kingdom)

OBJECTIVE: To assess the early variation of clinical periodontal indexes and Porphyronwnas gingivulis (Pgingivalis) in subgingival plaques of orthodontic adults. METHODS: 11 orthodontic adults were selected. Clinical periodontal indexes of observed teeth were examined at three different time points: Before orthodontic treatment, the first month after treatment and the third month after treatment, and subgingival plaques were collected simultaneously at each time point. Clinical periodontal indexes included four ones: Plaque index (PU), sulcus bleeding index (SBI), probing depth (PD), and attachment loss. We used real-time quantitative polymerase chain reaction to detect and quantitate the number of total bacteria and P.gingivalis in each sample to obtain positive rate of P.gingivalis and the percentage of P. gingiaalis in total bacteria. RESULTS: PLI and SBI of the first and third month were more than that of the baseline (P < 0.05). PD rose at the first month after treatment (P < 0.05), and then dropped at the third month. PD of all the 11 participants was lower than 2 mm. No attachment loss was found. The positive rate of P.gingivolis was stable (45.5%) and the proportion of Pgingivalis had no significant difference (p > 0.05) at each time point. CONCLUSION: Fixed orthodontic appliances caused plaque accumulation, accordingly slight gingiva inflammation and the increasement of P.gingivalis occurred in the early stage, hut none periodontitis was found.

Lu H; Zhou HM; Song XB; Sun JL; Liu HY

2010-04-01

 
 
 
 
161

Nucleotide sequence of the Porphyromonas gingivalis W83 recA homolog and construction of a recA-deficient mutant.  

UK PubMed Central (United Kingdom)

Degenerate oligonucleotide primers were used in PCR to amplify a region of the recA homolog from Porphyromonas gingivalis W83. The resulting PCR fragment was used as a probe to identify a recombinant lambda DASH phage (L10) carrying the P. gingivalis recA homolog. The recA homolog was localized to a 2.1-kb BamHI fragment. The nucleotide sequence of this 2.1-kb fragment was determined, and a 1.02-kb open reading frame (341 amino acids) was detected. The predicted amino acid sequence was strikingly similar (90% identical residues) to the RecA protein from Bacteroides fragilis. No SOS box, characteristic of LexA-regulated promoters, was found in the 5' upstream region of the P. gingivalis recA homolog. In both methyl methanesulfonate and UV survival experiments the recA homolog from P. gingivalis complemented the recA mutation of Escherichia coli HB101. The cloned P. gingivalis recA gene was insertionally inactivated with the ermF-ermAM antibiotic resistance cassette to create a recA-deficient mutant (FLL33) by allelic exchange. The recA-deficient mutant was significantly more sensitive to UV irradiation than the wild-type strain, W83. W83 and FLL33 showed the same level of virulence in in vivo experiments using a mouse model. These results suggest that the recA gene in P. gingivalis W83 plays the expected role of repairing DNA damage caused by UV irradiation. However, inactivation of this gene did not alter the virulence of P. gingivalis in the mouse model.

Fletcher HM; Morgan RM; Macrina FL

1997-11-01

162

Nucleotide sequence of the Porphyromonas gingivalis W83 recA homolog and construction of a recA-deficient mutant.  

Science.gov (United States)

Degenerate oligonucleotide primers were used in PCR to amplify a region of the recA homolog from Porphyromonas gingivalis W83. The resulting PCR fragment was used as a probe to identify a recombinant lambda DASH phage (L10) carrying the P. gingivalis recA homolog. The recA homolog was localized to a 2.1-kb BamHI fragment. The nucleotide sequence of this 2.1-kb fragment was determined, and a 1.02-kb open reading frame (341 amino acids) was detected. The predicted amino acid sequence was strikingly similar (90% identical residues) to the RecA protein from Bacteroides fragilis. No SOS box, characteristic of LexA-regulated promoters, was found in the 5' upstream region of the P. gingivalis recA homolog. In both methyl methanesulfonate and UV survival experiments the recA homolog from P. gingivalis complemented the recA mutation of Escherichia coli HB101. The cloned P. gingivalis recA gene was insertionally inactivated with the ermF-ermAM antibiotic resistance cassette to create a recA-deficient mutant (FLL33) by allelic exchange. The recA-deficient mutant was significantly more sensitive to UV irradiation than the wild-type strain, W83. W83 and FLL33 showed the same level of virulence in in vivo experiments using a mouse model. These results suggest that the recA gene in P. gingivalis W83 plays the expected role of repairing DNA damage caused by UV irradiation. However, inactivation of this gene did not alter the virulence of P. gingivalis in the mouse model. PMID:9353038

Fletcher, H M; Morgan, R M; Macrina, F L

1997-11-01

163

Prolyl tripeptidyl peptidase from Porphyromonas gingivalis. A novel enzyme with possible pathological implications for the development of periodontitis.  

Science.gov (United States)

Porphyromonas gingivalis possesses a complex proteolytic system, which is essential for both its growth and evasion of host defense mechanisms. In this report we characterized, both at a protein and genomic level, a novel peptidase of this system with prolyl tripeptidyl peptidase activity. The enzyme was purified to homogeneity, and its enzymatic activity and biochemical properties were investigated. The amino acid sequence at the amino terminus and of internal peptide fragments enabled identification of the gene encoding this enzyme, which we refer to as PtpA for prolyl tripeptidyl peptidase A. The gene encodes an 82-kDa protein, which contains a GWSYGG motif, characteristic for members of the S9 prolyl oligopeptidase family of serine proteases. However, it does not share any structural similarity to other tripeptidyl peptidases, which belong to the subtilisin family. The production of prolyl tripeptidyl peptidase may contribute to the pathogenesis of periodontal tissue destruction through the mutual interaction of this enzyme, host and bacterial collagenases, and dipeptidyl peptidases in the degradation of collagen during the course of infection. PMID:10092598

Banbula, A; Mak, P; Bugno, M; Silberring, J; Dubin, A; Nelson, D; Travis, J; Potempa, J

1999-04-01

164

Prolyl tripeptidyl peptidase from Porphyromonas gingivalis. A novel enzyme with possible pathological implications for the development of periodontitis.  

UK PubMed Central (United Kingdom)

Porphyromonas gingivalis possesses a complex proteolytic system, which is essential for both its growth and evasion of host defense mechanisms. In this report we characterized, both at a protein and genomic level, a novel peptidase of this system with prolyl tripeptidyl peptidase activity. The enzyme was purified to homogeneity, and its enzymatic activity and biochemical properties were investigated. The amino acid sequence at the amino terminus and of internal peptide fragments enabled identification of the gene encoding this enzyme, which we refer to as PtpA for prolyl tripeptidyl peptidase A. The gene encodes an 82-kDa protein, which contains a GWSYGG motif, characteristic for members of the S9 prolyl oligopeptidase family of serine proteases. However, it does not share any structural similarity to other tripeptidyl peptidases, which belong to the subtilisin family. The production of prolyl tripeptidyl peptidase may contribute to the pathogenesis of periodontal tissue destruction through the mutual interaction of this enzyme, host and bacterial collagenases, and dipeptidyl peptidases in the degradation of collagen during the course of infection.

Banbula A; Mak P; Bugno M; Silberring J; Dubin A; Nelson D; Travis J; Potempa J

1999-04-01

165

Oral Porphyromonas gingivalis translocates to liver and regulates hepatic glycogen synthesis through the Akt/GSK-3? signaling pathway.  

UK PubMed Central (United Kingdom)

Periodontal diseases are common chronic inflammatory disorders that result in the destruction of tissues around teeth. Many clinical studies suggest that periodontal diseases are risk factors for insulin resistance and diabetic mellitus development. However, the molecular mechanisms by which periodontal diseases regulate the progress of diabetes mellitus remain unknown. In this study, we investigated whether Porphyromonas gingivalis (P. g.), a major pathogen of periodontal diseases, present in the oral cavity, moves to the liver and affects hepatic glycogen synthesis. SNAP26b-tagged P.g. (SNAP-P.g.) was introduced into the oral cavity to induce periodontal disease in 4-week old female Balb/c mice. SNAP-P.g. was detected in liver extracted from SNAP-P.g.-treated mice using nested PCR analysis. High blood glucose levels tended to promote SNAP-P.g. translocation from the oral cavity to the liver in mice. Periodic acid-Schiff staining suggested that hepatic glycogen synthesis decreased in SNAP-P.g-treated mice. SNAP-P.g. was also internalized into the human hepatoma cell line HepG2, and this attenuated the phosphorylation of insulin receptor substrate (IRS)-1, Akt and glycogen synthase kinase-3? induced by insulin. Insulin-induced glycogen synthesis was suppressed by SNAP-P.g. in HepG2 cells. Our results suggest that P.g. translocation from the oral cavity to the liver may contribute to the progress of diabetes mellitus by influencing hepatic glycogenesis.

Ishikawa M; Yoshida K; Okamura H; Ochiai K; Takamura H; Fujiwara N; Ozaki K

2013-07-01

166

Evaluation of chemical composition and efficacy of Chinese propolis extract on Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans: An in vitro study  

Science.gov (United States)

Background: Propolis as a natural remedy has maintained its popularity over long periods of time. The aim of this study was to determine the chemical composition in terms of total phenolic compounds and flavonoids present in Chinese propolis and to carry out an in vitro evaluation of its antimicrobial activity and the minimal inhibitory concentrations for Porphyromonas gingivalis (Pg) and Aggregatibacter actinomycetemcomitans (Aa). Materials and Methods: From the ethanolic extract of propolis (EEP), total phenol content was determined by the Folin–Ciocalteau method, flavones and flavonols by the modified aluminum chloride colorimetric method, and flavanones by the 2.4-dinitrophenylhydrazine (2,4-DNP) method. Agar well diffusion assay was used to evaluate the antimicrobial potential of propolis against Pg and Aa. The minimum inhibitory concentration of propolis against the two bacteria was determined using serial tube dilution technique. Results: The total concentration of phenol in the EEP was 19.44%, flavones and flavonols 2.616%, and flavanones 16.176%. The inhibitory zone depicting antimicrobial activity ranged from 18 to 25 mm for Pg and from 12 to 14 mm for Aa. The concentration range of Chinese propolis that is sensitive to inhibit the growth of Pg was 0.1–0.0125 ?g/ml and for Aa it was 0.1–0.025 ?g/ml. Conclusion: These data suggest that Chinese propolis has potent antimicrobial activity against the two periodontopathogens, suggesting its possible use as a natural alternative to the widely used synthetic antibiotics for periodontal therapy.

Agarwal, Garima; Vemanaradhya, Gayathri G.; Mehta, Dhoom S.

2012-01-01

167

A Computational Simulation Study of Benzamidine Derivatives Binding to Arginine-Specific Gingipain (HRgpA) from Periodontopathogen Porphyromonas gingivalis  

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Full Text Available We have shown that the binding free energy calculation from molecular dynamics can be adapted successfully to cysteine proteinases, such as arginine-specific gingipain (HRgpA) from Porphyromonas gingivalis. The binding free energy obtained is in good agreement with the available experimental data for eight benzamidine derivatives including urea and ether linker. The calculations showed that the electrostatic energies between HRgpA and inhibitors were important in determining the relative affinities of the inhibitors to the HRgpA, with an average binding free energy of about ?5 kcal/mol. The average structures of the eight complexes suggest that benzamidine inhibitors interact with Asp387, His435, and Cys468 by hydrogen bonding and with Trp508 by hydrophilic interactions that are essential for the activities of benzamidine inhibitors. It can therefore be expected that the method provides a reliable tool for the investigation of new HRgpA inhibitors. This finding could significantly benefit the future design of HRgpA inhibitors.

Dooil Kim; Dae-Sil Lee

2010-01-01

168

Chlorhexidine varnishes effectively inhibit Porphyromonas gingivalis and Streptococcus mutans - An in vivo study  

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Full Text Available Background: Chlorhexidine varnish (Cervitec- Ivoclar Vivadent- Liechtenstein) is a sustained-release delivery system that can provide protection against white spots and gingivitis, which are common iatrogenic side effects of orthodontic treatment. Chlorhexidine in varnish form does not depend on patient compliance, does not stain teeth or alter taste sensation like the mouth rinse. Materials and Methods : A split-mouth technique was followed in the treatment of 30 patients selected by stringent selection criteria, evaluating a single application of the test varnish on two randomly allotted quadrants along with a placebo on the other two quadrants. Streptococcus mutans counts responsible for white spots and P. gingivalis count [using PCR test] responsible for gingivitis were done at the start of the study, and then 1 and 3 months later. Results: The chlorhexidine varnish reduced the Streptococci mutans count at the end of 1 month, and this reduction was statistically significant. At the end of 3 months, there was no difference in the S. mutans counts between the two groups. There was a statistically significant reduction in the P. gingivalis count at the end of both 1 and 3 months in comparison to the placebo group. Conclusion: Chlorhexidine varnishes are capable of reducing S. mutans and P. gingivalis and gingivitis, thus improving the overall oral health of the patient. The side effects of chlorhexidine mouth rinses are not seen with this varnish. An application schedule of at least once a month is recommended as the effectiveness is reduced comparatively at the end of 3 months.

George Ashwin; Kalangi Suresh; Vasudevan Mithuna; Krishnaswamy N

2010-01-01

169

Temporal activation of anti- and pro-apoptotic factors in human gingival fibroblasts infected with the periodontal pathogen, Porphyromonas gingivalis: potential role of bacterial proteases in host signalling  

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Full Text Available Abstract Background Porphyromonas gingivalis is the foremost oral pathogen of adult periodontitis in humans. However, the mechanisms of bacterial invasion and the resultant destruction of the gingival tissue remain largely undefined. Results We report host-P. gingivalis interactions in primary human gingival fibroblast (HGF) cells. Quantitative immunostaining revealed the need for a high multiplicity of infection for optimal infection. Early in infection (2–12 h), P. gingivalis activated the proinflammatory transcription factor NF-kappa B, partly via the PI3 kinase/AKT pathway. This was accompanied by the induction of cellular anti-apoptotic genes, including Bfl-1, Boo, Bcl-XL, Bcl2, Mcl-1, Bcl-w and Survivin. Late in infection (24–36 h) the anti-apoptotic genes largely shut down and the pro-apoptotic genes, including Nip3, Hrk, Bak, Bik, Bok, Bax, Bad, Bim and Moap-1, were activated. Apoptosis was characterized by nuclear DNA degradation and activation of caspases-3, -6, -7 and -9 via the intrinsic mitochondrial pathway. Use of inhibitors revealed an anti-apoptotic function of NF-kappa B and PI3 kinase in P. gingivalis-infected HGF cells. Use of a triple protease mutant P. gingivalis lacking three major gingipains (rgpA rgpB kgp) suggested a role of some or all these proteases in myriad aspects of bacteria-gingival interaction. Conclusion The pathology of the gingival fibroblast in P. gingivalis infection is affected by a temporal shift from cellular survival response to apoptosis, regulated by a number of anti- and pro-apoptotic molecules. The gingipain group of proteases affects bacteria-host interactions and may directly promote apoptosis by intracellular proteolytic activation of caspase-3.

Urnowey Sonya; Ansai Toshihiro; Bitko Vira; Nakayama Koji; Takehara Tadamichi; Barik Sailen

2006-01-01

170

Nasal immunization with the 40-kDa outer membrane protein of Porphyromonas gingivalis plus cholera toxin induces protective immunity in aged mice.  

UK PubMed Central (United Kingdom)

Previous studies have demonstrated that nasal administration of the outer membrane protein of Porphyromonas gingivalis (40k-OMP) with cholera toxin (CT) as an adjuvant elicits protective immune responses against P. gingivalis in young mice. In the present study, we investigated whether administration of 40k-OMP plus CT would also induce 40k-OMP-specific antibody (Ab) responses to provide protective immunity against P. gingivalis infection in aged mice. Nasal immunization with 40k-OMP plus CT elicited 40k-OMP-specific IgG and IgA Ab responses in serum and a significant anti-40k-OMP IgA Ab response in nasal washes and saliva in 1- and 2-year-old mice. Furthermore, both Th1- and Th2-type cytokine responses were induced by the immunization, and cytokine-associated IgG1, IgG2a, and IgG2b Ab responses were observed in the spleens of aged mice. Although the aged mice showed lower 40k-OMP-specific Ab responses than young mice, their mucosal IgA Ab titers as well as serum IgG Ab responses indicated a retained ability to mediate protective immunity; the only exception was saliva in 2-year-old mice. These findings suggest that nasal immunization with 40k-OMP plus CT can be a potential vaccination strategy for eliciting levels of Abs sufficient to provide protective immunity against P. gingivalis infection in aged mice.

Cai Y; Kurita-Ochiai T; Kobayashi R; Hashizume T; Yamamoto M

2013-01-01

171

The sinR ortholog PGN_0088 encodes a transcriptional regulator that inhibits polysaccharide synthesis in Porphyromonas gingivalis ATCC 33277 biofilms.  

UK PubMed Central (United Kingdom)

Biofilm-forming cells are distinct from well characterized planktonic cells and aggregate in the extracellular matrix, the so-called extracellular polymeric substances (EPS). The sinR gene of Bacillus subtilis encodes a transcriptional regulator that is known to be involved in the biosynthesis of EPS in biofilms. Porphyromonas gingivalis inhabits the subgingival and extraradicular biofilm of humans and is one of the primary pathogens that cause progressive marginal and refractory apical periodontitis. Furthermore, P. gingivalis possesses PGN_0088, which encodes a putative ortholog of B. subtilis sinR. Here, we investigated the role of PGN_0088 (sinR) on biofilm formation. P. gingivalis strains formed biofilms on saliva-coated glass surfaces in phosphate buffered saline. Quantitative analysis indicated that the biofilm of the sinR null mutant consisted of dense exopolysaccharide. Microscopic observations showed that the increased levels of exopolysaccharide produced by the sinR mutant changed the morphology of the EPS to a mesh-liked structure. Furthermore, physical analyses suggested that the enrichment of exopolysaccharide in the EPS enhanced the resistance of the biofilm to hydrodynamic shear force. The results presented here demonstrate sinR plays important roles in the ability of P. gingivalis strain ATCC 33277 to act as a negative mediator of exopolysaccharide accumulation and is indirectly associated with the structure of the EPS and the force of its adhesion to surfaces.

Yamamoto R; Noiri Y; Yamaguchi M; Asahi Y; Maezono H; Kuboniwa M; Hayashi M; Ebisu S

2013-01-01

172

Effects of Japanese traditional herbal medicines (Kampo) on growth and virulence properties of Porphyromonas gingivalis and viability of oral epithelial cells.  

Science.gov (United States)

Abstract Context: Kampos, commonly used in Japanese traditional medicine, are standardized herbal mixtures that have been used for centuries to treat a variety of ailments. We hypothesized that Kampos may have unidentified properties that may be beneficial in periodontitis, an inflammatory disease affecting the tooth-supporting tissues. Objective: The aim of our study was to investigate various Kampos and their natural ingredients for their effects on Porphyromonas gingivalis growth, adherence to epithelial cells and proteinase activity. In addition, their effects on oral epithelial cell viability were evaluated. Materials and methods: Growth inhibition of P. gingivalis by various Kampos and their natural ingredients was evaluated by a microdilution broth assay method. Their effects on P. gingivalis proteinase activity and adherence to oral epithelial cells were determined by fluorometric assays. The cytotoxicity of test compounds towards oral epithelial cells was evaluated by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] test. Results: Of the 27 Kampos tested, 7 were found to inhibit the growth of P. gingivalis. The lowest minimal inhibitory concentration (MIC) (250?µg/ml) was obtained with TJ-113. Analysis of the composition of the seven active Kampos showed that they contain Chinese rhubarb as a common ingredient. Therefore, additional growth inhibitory assays on P. gingivalis were carried out with purified anthraquinones known to be present in rhubarb. Aloe-emodin and rhein possessed the strongest antibacterial effects towards P. gingivalis with an MIC of 0.78?µg/ml. The seven Kampos containing rhubarb and purified anthraquinones also exhibited the capacity to decrease the adherence of P. gingivalis to oral epithelial cells and to reduce its proteinase activity. The most important anti-adherence effect of Kampo was obtained with TJ-126; at 250?µg/ml it reduced adherence of P. gingivalis to epithelial cells by 83%. Purified anthraquinones were found to be less active than Kampos. Kampo TJ-113 was found to be the most effective for inhibition of gelatin degradation (49% inhibition at 62.5?µg/ml). Again, purified anthraquinones inhibited gelatin degradation to a lesser extent. Lastly, none of the tested compounds showed cytotoxicity towards oral epithelial cells at the effective concentrations. Conclusion: Kampos containing rhubarb and its anthraquinone derivatives may represent promising molecules for controlling periodontal diseases through their capacity to inhibit P. gingivalis growth and virulence properties. PMID:23987742

Liao, James; Zhao, Lei; Yoshioka, Masami; Hinode, Daisuke; Grenier, Daniel

2013-08-29

173

Effects of Japanese traditional herbal medicines (Kampo) on growth and virulence properties of Porphyromonas gingivalis and viability of oral epithelial cells.  

UK PubMed Central (United Kingdom)

Abstract Context: Kampos, commonly used in Japanese traditional medicine, are standardized herbal mixtures that have been used for centuries to treat a variety of ailments. We hypothesized that Kampos may have unidentified properties that may be beneficial in periodontitis, an inflammatory disease affecting the tooth-supporting tissues. Objective: The aim of our study was to investigate various Kampos and their natural ingredients for their effects on Porphyromonas gingivalis growth, adherence to epithelial cells and proteinase activity. In addition, their effects on oral epithelial cell viability were evaluated. Materials and methods: Growth inhibition of P. gingivalis by various Kampos and their natural ingredients was evaluated by a microdilution broth assay method. Their effects on P. gingivalis proteinase activity and adherence to oral epithelial cells were determined by fluorometric assays. The cytotoxicity of test compounds towards oral epithelial cells was evaluated by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] test. Results: Of the 27 Kampos tested, 7 were found to inhibit the growth of P. gingivalis. The lowest minimal inhibitory concentration (MIC) (250?µg/ml) was obtained with TJ-113. Analysis of the composition of the seven active Kampos showed that they contain Chinese rhubarb as a common ingredient. Therefore, additional growth inhibitory assays on P. gingivalis were carried out with purified anthraquinones known to be present in rhubarb. Aloe-emodin and rhein possessed the strongest antibacterial effects towards P. gingivalis with an MIC of 0.78?µg/ml. The seven Kampos containing rhubarb and purified anthraquinones also exhibited the capacity to decrease the adherence of P. gingivalis to oral epithelial cells and to reduce its proteinase activity. The most important anti-adherence effect of Kampo was obtained with TJ-126; at 250?µg/ml it reduced adherence of P. gingivalis to epithelial cells by 83%. Purified anthraquinones were found to be less active than Kampos. Kampo TJ-113 was found to be the most effective for inhibition of gelatin degradation (49% inhibition at 62.5?µg/ml). Again, purified anthraquinones inhibited gelatin degradation to a lesser extent. Lastly, none of the tested compounds showed cytotoxicity towards oral epithelial cells at the effective concentrations. Conclusion: Kampos containing rhubarb and its anthraquinone derivatives may represent promising molecules for controlling periodontal diseases through their capacity to inhibit P. gingivalis growth and virulence properties.

Liao J; Zhao L; Yoshioka M; Hinode D; Grenier D

2013-08-01

174

Antimicrobial photodynamic therapy using a diode laser with a potential new photosensitizer, indocyanine green-loaded nanospheres, may be effective for the clearance of Porphyromonas gingivalis.  

UK PubMed Central (United Kingdom)

BACKGROUND: Antimicrobial photodynamic therapy (aPDT) is a new treatment method for the removal of infectious pathogens using a photosensitizer and light of a specific wavelength, e.g., toluidine blue with a wavelength of about 600 nm. We explored a new photosensitizer and focused on indocyanine green (ICG), which has high absorption at a wavelength of 800-805 nm. We investigated the bactericidal effect of PDT on Porphyromonas gingivalis using a new photosensitizer, ICG-loaded nanospheres with an 805 nm wavelength low-level diode laser irradiation. METHODS: We designed ICG-loaded nanospheres coated with chitosan (ICG-Nano/c) as a photosensitizer. A solution containing Porphyromonas gingivalis (10(8)  CFU/mL) with or without ICG-Nano/c (or ICG) was prepared and irradiated with a diode laser or without laser irradiation as a negative control. The irradiation settings were 0.5 W with a duty ratio of 10%, for 3-100 ms in repeated pulse (RPT) or continuous wave mode. CFU were counted after 7 d of anaerobic culture. RESULTS: We observed that ICG-Nano/c could adhere to the surface of P. gingivalis. When ICG-Nano/c was used for aPDT, irradiation with RPT 100 ms mode gave the lowest increase in temperature. Laser irradiation with ICG-Nano/c significantly reduced the number of P. gingivalis (i.e., approximately 2-log10 bacterial killing). The greatest bactericidal effect was found in the RPT 100 ms group. However, laser irradiation (RPT 100 ms) with ICG, as well as without photosensitizer, had no effect on the number of bacteria. CONCLUSIONS: Within the limits of this study, ICG-Nano/c with low-level diode laser (0.5 W; 805 nm) irradiation showed an aPDT-like effect, which might be useful for a potential photodynamic periodontal therapy.

Nagahara A; Mitani A; Fukuda M; Yamamoto H; Tahara K; Morita I; Ting CC; Watanabe T; Fujimura T; Osawa K; Sato S; Takahashi S; Iwamura Y; Kuroyanagi T; Kawashima Y; Noguchi T

2013-10-01

175

Association between clinical parameters and the presence of Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis in patients with progressive periodontal lesions  

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Full Text Available Background/Aim. Periodontitis is a chronic inflammatory disease of periodontal tissues with consequential is bone loss as a result of host immunological reactions caused by periopathogens. The aim of the study was to investigate if there is a correlation between clinical parameters and the presence of two most aggressive periopathogens (Aggregatibacter actinomycetemcomitans - Aa and Porphyromonas gingivalis - Pg) in patients with progressive periodontal lesions. Methods. A total of 34 systemic healthy people, 23 to 70 years old, were included in the study. The patients were clinically and radiologically examined, and after that, the representative pocket with greatest pocket depth was chosen and the sample was collected from that place. The measured clinic parameters were: gingival index, index of gingival bleeding, pocket depth and plaque indices. The multiplex Polymerase Chain Reaction (PCR) method was used for detection of periopathogens. After obtaining results, appropriate statistical tests were used to correlate the clinical and microbiological results. Results. Aa and Pg were detected in the same percentage of samples. Aa and Pg were detected in 35.29% samples alone, and in 29.41% both were detected. The values of measured clinical parameters did not show a statistical significance between the groups. In analysis of correlations among clinical parameters inside the groups, a statistical significance was found only between gingival and plaque index in the group with Aa. Conclusion. Clinical course of periodontitis in the developed stage does not differ in relation to the presence of different periopathogens as the major inductors of immunologically guided destructive processes.

Raki? Mia; Zeli? Ksenija; Pavlica Dušan; Hadžimihajlovi? Miloš; Milašin Jelena; Mili?i? Biljana; Nikoli? Nebojša; Stamatovi? Novak; Mati? Smiljana; Aleksi? Zoran; Jankovi? Saša

2010-01-01

176

Nasal Vaccination with the 40-Kilodalton Outer Membrane Protein of Porphyromonas gingivalis and a Nontoxic Chimeric Enterotoxin Adjuvant Induces Long-Term Protective Immunity with Reduced Levels of Immunoglobulin E Antibodies?  

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In this study, we demonstrated that the 40-kDa outer membrane protein of Porphyromonas gingivalis (40-kDa OMP) nasally administered with a nontoxic chimeric adjuvant that combines the A subunit of mutant cholera toxin E112K with the pentameric B subunit of heat-labile enterotoxin from enterotoxigeni...

Momoi, Fumiki; Hashizume, Tomomi; Kurita-Ochiai, Tomoko; Yuki, Yoshikazu; Kiyono, Hiroshi; Yamamoto, Masafumi

177

Comparison of 1-Periodontal Indices and Cultural Porphyromonas Gingivalis Colony Count in Aggressive Periodontitis Patients Treated by Scaling and Rootplanning with or Without Metronidazole Gel  

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Full Text Available Objective: Systemic antibiotics and locally applied antimicrobial agents have been suggested to enhance clinical parameters. Patients exhibiting aggressive periodontitis in particular benefit from adjunctive antibiotic therapy. The purpose of this investigation was to evaluate the effect of local antibiotic therapy with metronidazoleadjunctively to scaling and root planning (SRP) in the treatment of aggressive periodontitis.Materials and Methods: Twenty patients diagnosed with aggressive periodontitis were placed in a spilt mouth design. Microbial specimens were taken from thedeepest pocket of the teeth. The sites that had positive results of Porphyromonas gingivalis (P.g) were located randomly to receive SRP treatment in the control group and SRP plus metronidazole gel in the test group. Pocket probing depth (PPD), clinical attachment level (CAL) and bleeding on probing (BOP) parameters and numbers of P.g. colony were taken at baseline, 6 weeks and 12 weeks later.All data were collected and analyzed and tested by Wilcoxon and paired t test. Results: The case group patients had significantly better results in BOP, PPD and the number of P.g colony count reduction in comparison with the control group (p0.05).Conclusion: In non-surgical periodontal treatment of aggressive periodontits adjunctive metronidazole gel therapy has a better effect on the reduction of porphyromonas gingivalis content of pockets.

Z. Kadkhoda; S. Rafiei Tari; P. Owlia; S. Seyyed Zadeh Sabounchei

2012-01-01

178

The vimE gene downstream of vimA is independently expressed and is involved in modulating proteolytic activity in Porphyromonas gingivalis W83.  

UK PubMed Central (United Kingdom)

Regulation/activation of the Porphyromonas gingivalis gingipains is poorly understood. A unique 1.3-kb open reading frame downstream of the bcp-recA-vimA transcriptional unit was cloned, insertionally inactivated with the ermF-ermAM antibiotic resistance cassette, and used to create a defective mutant by allelic exchange. In contrast to the wild-type W83 strain, the growth rate of the mutant strain (designated FLL93) was reduced, and when plated on Brucella blood agar it was nonpigmented and nonhemolytic. Arginine- and lysine-specific gingipain activities were reduced by approximately 90 and 85%, respectively, relative to activities of the parent strain. These activities were unaffected by the culture's growth phase, in contrast to the vimA-defective mutant P. gingivalis FLL92, which has increased proteolytic activity in stationary phase. Expression of the rgpA, rgpB, and kgp gingipain genes was unaltered in P. gingivalis FLL93 compared to that of the wild-type strain. Further, in extracellular protein fractions a 64-kDa band was identified that was immunoreactive with the RgpB-specific proenzyme antibodies. Active-site labeling with dansyl-glutamyl-glycyl-arginyl chloromethyl ketone or immunoblot analysis showed no detectable protein band representing the gingipain catalytic domain. In vitro protease activity could be slightly induced by a urea denaturation-renaturation cycle in an extracellular protein fraction, in contrast to the vimA-defective mutant P. gingivalis FLL92. Expression of flanking genes, including recA, vimA, and Pg0792, was unaltered by the mutation. Taken together, these results suggest that the vimA downstream gene, designated vimE (for virulence-modulating gene E), is involved in the regulation of protease activity in P. gingivalis.

Vanterpool E; Roy F; Fletcher HM

2004-10-01

179

The vimE gene downstream of vimA is independently expressed and is involved in modulating proteolytic activity in Porphyromonas gingivalis W83.  

Science.gov (United States)

Regulation/activation of the Porphyromonas gingivalis gingipains is poorly understood. A unique 1.3-kb open reading frame downstream of the bcp-recA-vimA transcriptional unit was cloned, insertionally inactivated with the ermF-ermAM antibiotic resistance cassette, and used to create a defective mutant by allelic exchange. In contrast to the wild-type W83 strain, the growth rate of the mutant strain (designated FLL93) was reduced, and when plated on Brucella blood agar it was nonpigmented and nonhemolytic. Arginine- and lysine-specific gingipain activities were reduced by approximately 90 and 85%, respectively, relative to activities of the parent strain. These activities were unaffected by the culture's growth phase, in contrast to the vimA-defective mutant P. gingivalis FLL92, which has increased proteolytic activity in stationary phase. Expression of the rgpA, rgpB, and kgp gingipain genes was unaltered in P. gingivalis FLL93 compared to that of the wild-type strain. Further, in extracellular protein fractions a 64-kDa band was identified that was immunoreactive with the RgpB-specific proenzyme antibodies. Active-site labeling with dansyl-glutamyl-glycyl-arginyl chloromethyl ketone or immunoblot analysis showed no detectable protein band representing the gingipain catalytic domain. In vitro protease activity could be slightly induced by a urea denaturation-renaturation cycle in an extracellular protein fraction, in contrast to the vimA-defective mutant P. gingivalis FLL92. Expression of flanking genes, including recA, vimA, and Pg0792, was unaltered by the mutation. Taken together, these results suggest that the vimA downstream gene, designated vimE (for virulence-modulating gene E), is involved in the regulation of protease activity in P. gingivalis. PMID:15385452

Vanterpool, Elaine; Roy, Francis; Fletcher, Hansel M

2004-10-01

180

Identification of a Gingipain-Sensitive Surface Ligand of Porphyromonas gingivalis That Induces Toll-Like Receptor 2- and 4-Independent NF-?B Activation in CHO Cells?  

Science.gov (United States)

Porphyromonas gingivalis is a major periodontal pathogen that has the pathogenic proteinases Arg-specific gingipain and Lys-specific gingipain. We previously found that a cell surface component on P. gingivalis is able to induce Toll-like receptor 2 (TLR2)- and TLR4-independent signaling in 7.19 cells and that this component can be degraded by gingipains. In this study, we purified this component from the P. gingivalis gingipain-null mutant KDP136 and obtained two candidate proteins. Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry analysis showed that the proteins, with molecular masses of 123 and 43 kDa, were encoded by PGN_0748 and PGN_0728 (pgm6), respectively, in the P. gingivalis ATCC 33277 genome sequence. The PGN_0748-encoded protein, which we refer to as gingipain-sensitive ligand A (GslA), reacted with antiserum that could effectively inhibit the activity of KDP136 to induce NF-?B activation in 7.19 cells, but Pgm6 did not. To further determine what protein is responsible for the NF-?B activation, we constructed gslA, pgm6, and pgm6 pgm7 deletion mutants from KDP136. When 7.19 cells were exposed to those mutants, the gslA deletion mutant did not induce NF-?B activation, whereas the pgm6 and pgm6 pgm7 deletion mutants did. Furthermore, NF-?B activation in 7.19 cells induced by KDP136 was partially inhibited by antiserum against a recombinant protein expressed from the 5?-terminal third of gslA. These results indicate that GslA is one of the factors that induce NF-?B activation in 7.19 cells. Interestingly, the gslA gene was present in four of seven P. gingivalis strains tested. This restricted distribution might be associated with the virulence potential of each strain.

Haruyama, Koki; Yoshimura, Atsutoshi; Naito, Mariko; Kishimoto, Mami; Shoji, Mikio; Abiko, Yoshimitsu; Hara, Yoshitaka; Nakayama, Koji

2009-01-01

 
 
 
 
181

Identification of proteinaceous inhibitors of a cysteine proteinase (an Arg-specific gingipain) from Porphyromonas gingivalis in rice grain, using targeted-proteomics approaches.  

UK PubMed Central (United Kingdom)

Porphyromonas gingivalis is known to be a major etiologic agent in the onset and progression of chronic periodontitis. Among various virulence factors that this bacterium produces, Arg- and Lys-specific cysteine proteinases (gingipains) are believed to be major determinants of the pathogenicity of P. gingivalis. Here, we report on our finding that there are inhibitors of these cysteine proteinases in a rice protein fraction. Comprehensive affinity chromatography and MS analyses resulted in the identification of 17 Arg-gingipain (Rgp)-interacting proteins in the rice endosperm. Of these, four proteins (i.e., a 26 kDa globulin, a plant lipid transfer/trypsin-alpha amylase inhibitor, the RA17 seed allergen, and an alpha amylase/trypsin inhibitor) were estimated to account for 90% of the Rgp inhibitory activity in the rice protein fraction, using a two-dimensional gel system of double-layer reverse zymography. In addition, a synthetic peptide derived from an Rgp-interacting protein, cyanate hydratase, could inhibit the growth of P. gingivalis and showed inhibitory activity against both the Arg- and Lys-gingipains. These results suggest that these rice proteins may be useful as nutraceutical ingredients for the prevention and management of periodontal diseases.

Taiyoji M; Shitomi Y; Taniguchi M; Saitoh E; Ohtsubo S

2009-11-01

182

Identification of proteinaceous inhibitors of a cysteine proteinase (an Arg-specific gingipain) from Porphyromonas gingivalis in rice grain, using targeted-proteomics approaches.  

Science.gov (United States)

Porphyromonas gingivalis is known to be a major etiologic agent in the onset and progression of chronic periodontitis. Among various virulence factors that this bacterium produces, Arg- and Lys-specific cysteine proteinases (gingipains) are believed to be major determinants of the pathogenicity of P. gingivalis. Here, we report on our finding that there are inhibitors of these cysteine proteinases in a rice protein fraction. Comprehensive affinity chromatography and MS analyses resulted in the identification of 17 Arg-gingipain (Rgp)-interacting proteins in the rice endosperm. Of these, four proteins (i.e., a 26 kDa globulin, a plant lipid transfer/trypsin-alpha amylase inhibitor, the RA17 seed allergen, and an alpha amylase/trypsin inhibitor) were estimated to account for 90% of the Rgp inhibitory activity in the rice protein fraction, using a two-dimensional gel system of double-layer reverse zymography. In addition, a synthetic peptide derived from an Rgp-interacting protein, cyanate hydratase, could inhibit the growth of P. gingivalis and showed inhibitory activity against both the Arg- and Lys-gingipains. These results suggest that these rice proteins may be useful as nutraceutical ingredients for the prevention and management of periodontal diseases. PMID:19691286

Taiyoji, Mayumi; Shitomi, Yasuyuki; Taniguchi, Masayuki; Saitoh, Eiichi; Ohtsubo, Sadami

2009-11-01

183

Specific in situ visualization of plasma cells producing antibodies against Porphyromonas gingivalis in gingival radicular cyst: application of the enzyme-labeled antigen method.  

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The enzyme-labeled antigen method was applied to visualize plasma cells producing antibodies to Porphyromonas gingivalis, flora of the human oral cavity. Antibodies to P. gingivalis have reportedly been detected in sera of patients with periodontitis. Biotinylated bacterial antigens, Ag53, and four gingipain domains (Arg-pro, Arg-hgp, Lys-pro, and Lys-hgp) were prepared by the cell-free protein synthesis system using the wheat germ extract. In paraformaldehyde-fixed frozen sections of rat lymph nodes experimentally immunized with Ag53-positive and Ag53-negative P. gingivalis, plasma cells were labeled with biotinylated Arg-hgp and Lys-hgp. Antibodies to Ag53 were detected only in the nodes immunized with Ag53-positive bacteria. In two of eight lesions of gingival radicular cyst with inflammatory infiltration, CD138-positive plasma cells in frozen sections were signalized for Arg-hgp and Lys-hgp. An absorption study using unlabeled antigens confirmed the specificity of staining. The AlphaScreen method identified the same-type antibodies in tissue extracts but not in sera. Antibodies to Ag53, Arg-pro, and Lys-pro were undetectable. In two cases, serum antibodies to Arg-hgp and Lys-hgp were AlphaScreen positive, whereas plasma cells were scarcely observed within the lesions. These findings indicate the validity of the enzyme-labeled antigen method. This is the very first application of this novel histochemical technique to human clinical samples. PMID:21525188

Tsuge, Shinya; Mizutani, Yasuyoshi; Matsuoka, Kazuhiro; Sawasaki, Tatsuya; Endo, Yaeta; Naruishi, Koji; Maeda, Hiroshi; Takashiba, Shogo; Shiogama, Kazuya; Inada, Ken-Ichi; Tsutsumi, Yutaka

2011-04-27

184

Presencia de Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola y Aggregatibacter actinomycetemcomitans en el biofilm subgingival de pacientes diabéticos tipo 2: estudio transversal/ Presence of Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola and Aggregatibacter actinomycetemcomitans in the subgingival biofilm of diabetic mellitus 2 patients: a cross sectional study  

Scientific Electronic Library Online (English)

Full Text Available Abstract in spanish Antecedentes: La investigación de la microflora subgingival en pacientes diabéticos tipo 2 con periodontitis ha presentado resultados contradictorios. Objetivo: Determinar la presencia de Porphyromonas gingivalis, Tannerella forshytia, Treponema denticola y Aggregatibacter actinomycetemcomitans, en el biofilm subgingival de pacientes diabéticos tipo 2 y relacionarlo con el grado de control metabólico. Método: Estudio descriptivo transversal, en el cual se analizaron (more) 23 pacientes diabéticos derivados consecutivamente del Policlínico de Especialidades de la Universidad de los Andes. Previo consentimiento informado, se realizó un examen clínico periodontal que incluyó mediciones de profundidad al sondaje, nivel de inserción clínica y sangrado gingival. Fueron clasificados según severidad de periodontitis y control metabólico de la diabetes determinado por un promedio de 3 exámenes de hemoglobina glicosilada. La detección microbiológica se realizó mediante la técnica de reacción en cadena de la polimerasa. Resultados: En el grupo de pacientes estudiados, Treponema denticola y Tannerella forsythia fueron las bacterias más prevalentes (65.2%), seguida por Porphyromonas gingivalis (17.3%) y Aggregatibacter actinomycetemcomitans (13%). Los pacientes con peor control glicémico tuvieron una mayor presencia de Treponema denticola, Tannerella forsythia, Porphyromonas gingivalis y Agreggatibacter actinomycetemcomitans y un aumento en el índice de sangrado al sondaje. Conclusiones: En el grupo de pacientes diabéticos estudiado, las bacterias más prevalentes fueron Treponema denticola y Tannerella forsythia. Los pacientes diabéticos tipo 2 con moderado y mal control glicémico presentaron mayor presencia de los microorganismos estudiados, comparado con los grupos con mejores niveles de control glicémico. Abstract in english Background: The investigation of subgingival microflora in type 2 diabetic patients with periodontitis presented conflicting results. Aim: To determine the presence of Porphyromonas gingivalis, Tannerella forshytia, Treponema denticola and Aggregatibacter actinomycetemcomitans in subgingival biofilm of patients with diabetes type 2 and to relate it to the degree of metabolic control. Method: A descriptive study, which analyzed 23 diabetic patients consecutively referred f (more) rom the Internal Medicine Unit of Medicine Faculty at Universidad de los Andes was conducted. After obtaining an informed consent from the patients a clinical examination that included measurements of periodontal pocket depth, clinical attachment level and gingival bleeding was performed. The patients were classified according to the severity of periodontitis and metabolic control of diabetes as determined by an average of 3 of glycosylated haemoglobin tests. Microbial technique was performed by chain reaction of polymerase. Results: In the group of patients examined the most prevalent bacteria were, Treponema denticola and Tannerella forsythia (65.2%), followed by Porphyromonas gingivalis (17.3%) and Aggregatibacter actinomycetemcomitans (13%). Patients with poor glycemic control had a greater presence of Treponema denticola, Tannerella forsythia, Porphyromonas gingivalis and Agreggatibacter actinomycetemcomitans and an increase in the rate of bleeding on probing. Conclusions: In the group of diabetic patients studied, the most prevalent bacteria were Treponema denticola and Tannerella forsythia. Type 2 diabetic patients with moderate and poor glycemic control had a higher presence of these microorganisms, compared to groups with higher levels of glycemic control.

Quintero, AJ; Prada, P; Inostroza, CM; Chaparro, A; Sanz, AF; Ramírez, VL; Morales, HC

2011-08-01

185

Presencia de Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola y Aggregatibacter actinomycetemcomitans en el biofilm subgingival de pacientes diabéticos tipo 2: estudio transversal Presence of Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola and Aggregatibacter actinomycetemcomitans in the subgingival biofilm of diabetic mellitus 2 patients: a cross sectional study  

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Full Text Available Antecedentes: La investigación de la microflora subgingival en pacientes diabéticos tipo 2 con periodontitis ha presentado resultados contradictorios. Objetivo: Determinar la presencia de Porphyromonas gingivalis, Tannerella forshytia, Treponema denticola y Aggregatibacter actinomycetemcomitans, en el biofilm subgingival de pacientes diabéticos tipo 2 y relacionarlo con el grado de control metabólico. Método: Estudio descriptivo transversal, en el cual se analizaron 23 pacientes diabéticos derivados consecutivamente del Policlínico de Especialidades de la Universidad de los Andes. Previo consentimiento informado, se realizó un examen clínico periodontal que incluyó mediciones de profundidad al sondaje, nivel de inserción clínica y sangrado gingival. Fueron clasificados según severidad de periodontitis y control metabólico de la diabetes determinado por un promedio de 3 exámenes de hemoglobina glicosilada. La detección microbiológica se realizó mediante la técnica de reacción en cadena de la polimerasa. Resultados: En el grupo de pacientes estudiados, Treponema denticola y Tannerella forsythia fueron las bacterias más prevalentes (65.2%), seguida por Porphyromonas gingivalis (17.3%) y Aggregatibacter actinomycetemcomitans (13%). Los pacientes con peor control glicémico tuvieron una mayor presencia de Treponema denticola, Tannerella forsythia, Porphyromonas gingivalis y Agreggatibacter actinomycetemcomitans y un aumento en el índice de sangrado al sondaje. Conclusiones: En el grupo de pacientes diabéticos estudiado, las bacterias más prevalentes fueron Treponema denticola y Tannerella forsythia. Los pacientes diabéticos tipo 2 con moderado y mal control glicémico presentaron mayor presencia de los microorganismos estudiados, comparado con los grupos con mejores niveles de control glicémico.Background: The investigation of subgingival microflora in type 2 diabetic patients with periodontitis presented conflicting results. Aim: To determine the presence of Porphyromonas gingivalis, Tannerella forshytia, Treponema denticola and Aggregatibacter actinomycetemcomitans in subgingival biofilm of patients with diabetes type 2 and to relate it to the degree of metabolic control. Method: A descriptive study, which analyzed 23 diabetic patients consecutively referred from the Internal Medicine Unit of Medicine Faculty at Universidad de los Andes was conducted. After obtaining an informed consent from the patients a clinical examination that included measurements of periodontal pocket depth, clinical attachment level and gingival bleeding was performed. The patients were classified according to the severity of periodontitis and metabolic control of diabetes as determined by an average of 3 of glycosylated haemoglobin tests. Microbial technique was performed by chain reaction of polymerase. Results: In the group of patients examined the most prevalent bacteria were, Treponema denticola and Tannerella forsythia (65.2%), followed by Porphyromonas gingivalis (17.3%) and Aggregatibacter actinomycetemcomitans (13%). Patients with poor glycemic control had a greater presence of Treponema denticola, Tannerella forsythia, Porphyromonas gingivalis and Agreggatibacter actinomycetemcomitans and an increase in the rate of bleeding on probing. Conclusions: In the group of diabetic patients studied, the most prevalent bacteria were Treponema denticola and Tannerella forsythia. Type 2 diabetic patients with moderate and poor glycemic control had a higher presence of these microorganisms, compared to groups with higher levels of glycemic control.

AJ Quintero; P Prada; CM Inostroza; A Chaparro; AF Sanz; VL Ramírez; HC Morales

2011-01-01

186

Biological activities of lipopolysaccharides from oral bacteria and their relevance to the pathogenesis of chronic periodontitis.  

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Chronic periodontitis is a major cause of tooth loss in adults and is a consequence of the colonisation of the subgingival region by organisms such as Porphyromonas gingivalis, Prevotella intermedia and Fusobacterium nucleatum. Lipopolysaccharide (LPS) is a constituent of the cell walls of all of th...

Wilson, M

187

Antimicrobial effect of photodynamic therapy using high-power blue light-emitting diode and red-dye agent on Porphyromonas gingivalis.  

UK PubMed Central (United Kingdom)

BACKGROUND AND OBJECTIVE: Antimicrobial photodynamic therapy (a-PDT) using a combination of red-colored laser/light-emitting diode (LED) and blue dye has been employed for periodontal therapy and the antimicrobial effect seems promising. Blue light, which has favorable wavelength properties, would be more effective as a light source for a-PDT because blue light itself possesses an antimicrobial effect. This study aimed to investigate the effect of a-PDT using a novel combination of high-power blue LED and red-dye agent on Porphyromonas gingivalis in vitro. MATERIAL AND METHODS: Porphyromonas gingivalis ATCC 33277 suspension was irradiated with blue LED (BL) (425-470 nm) or red LED (RL) (625-635 nm) at 30-90 J/cm(2) , or was mixed with erythrosine (ER), phloxine B (PB) or rose bengal (RB) with or without BL irradiation (30 J/cm(2) ). RL (30 J/cm(2) ) in combination with toluidine blue was employed as positive control. All the suspensions of P. gingivalis were serially diluted, plated and incubated anaerobically, and the numbers of colony-forming units (CFUs) were counted on day 7. RESULTS: BL irradiation at 60 and 90 J/cm(2) demonstrated a significant reduction in the numbers of CFUs. ER, PB and RB solutions at 160 ?g/mL showed almost no or only a minimal reduction in the numbers of CFUs. BL at 30 J/cm(2) combined with ER, PB or RB at 160 ?g/mL resulted in a log reduction of 0.9, 1.0 and 7.1, respectively, in the numbers of CFUs; 30 J/cm(2) BL with RB at 1.6, 16 and 160 ?g/mL demonstrated a log reduction of 6.3, 8.0 and 5.5, respectively; and a log reduction of 5.2 was obtained after 30 J/cm(2) RL with 16 ?g/mL TB. CONCLUSION: Within the limits of this study, BL was found to have an antimicrobial/growth-inhibiting effect on P. gingivalis, and a-PDT using a combination of BL and RB shows promise as a new technical modality for bacterial elimination in periodontal therapy.

Chui C; Aoki A; Takeuchi Y; Sasaki Y; Hiratsuka K; Abiko Y; Izumi Y

2013-02-01

188

Isolation of the new anacardic acid 6-[16'Z-nonadecenyl]-salicylic acid and evaluation of its antimicrobial activity against Streptococcus mutans and Porphyromonas gingivalis.  

UK PubMed Central (United Kingdom)

A new anacardic acid, 6-[16'Z-nonadecenyl]-salicylic acid (1), along with seven known compounds, 6-[8'Z-pentadecenyl] salicylic acid (15:1 anacardic acid) (2), 6-nonadecenyl salicylic acid (anacardic acid 19:0) (3), 6-pentadecyl salicylic acid (anacardic acid 15:0) (4), masticadienonic acid (5), 3?-hydroxymasticadienonic acid (6), 3-epi-oleanolic acid (7) and ?-sitosterol, were isolated from the bark of Amphipterygium adstringens using a bioassay-guided fractionation method. The structure of the new compound (1) was elucidated by spectroscopic data interpretation. The known compounds (2-7) were identified by comparison of their spectroscopic data with reported values in the literature. Compounds 1-4 exhibited antibacterial activity against Streptococcus mutans and Porphyromonas gingivalis with minimum inhibitory concentrations ranging from 7 to 104?µg?mL and from 12 to 126?µg?mL, respectively.

Rivero-Cruz BE; Esturau N; Sánchez-Nieto S; Romero I; Castillo-Juárez I; Rivero-Cruz JF

2011-08-01

189

Isolation of the new anacardic acid 6-[16'Z-nonadecenyl]-salicylic acid and evaluation of its antimicrobial activity against Streptococcus mutans and Porphyromonas gingivalis.  

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A new anacardic acid, 6-[16'Z-nonadecenyl]-salicylic acid (1), along with seven known compounds, 6-[8'Z-pentadecenyl] salicylic acid (15:1 anacardic acid) (2), 6-nonadecenyl salicylic acid (anacardic acid 19:0) (3), 6-pentadecyl salicylic acid (anacardic acid 15:0) (4), masticadienonic acid (5), 3?-hydroxymasticadienonic acid (6), 3-epi-oleanolic acid (7) and ?-sitosterol, were isolated from the bark of Amphipterygium adstringens using a bioassay-guided fractionation method. The structure of the new compound (1) was elucidated by spectroscopic data interpretation. The known compounds (2-7) were identified by comparison of their spectroscopic data with reported values in the literature. Compounds 1-4 exhibited antibacterial activity against Streptococcus mutans and Porphyromonas gingivalis with minimum inhibitory concentrations ranging from 7 to 104?µg?mL and from 12 to 126?µg?mL, respectively. PMID:21815722

Rivero-Cruz, Blanca E; Esturau, Nuria; Sánchez-Nieto, Sobeida; Romero, Irma; Castillo-Juárez, Israel; Rivero-Cruz, J Fausto

2011-08-04

190

A highly catalytically active ?-carbonic anhydrase from the pathogenic anaerobe Porphyromonas gingivalis and its inhibition profile with anions and small molecules.  

UK PubMed Central (United Kingdom)

Carbonic anhydrases (CAs, EC 4.2.1.1) belonging to the ?-class are present in archaea, bacteria and plants but, except the Methanosarcina thermophila enzymes CAM and CAMH, they were poorly characterized so far. Here we report a new such enzyme (PgiCA), the ?-CA from the oral cavity pathogenic bacterium Porphyromonas gingivalis, the main causative agent of periodontitis. PgiCA showed a good catalytic activity for the CO2 hydration reaction, comparable to that of the human (h) isoform hCA I. Inorganic anions such as thiocyanate, cyanide, azide, hydrogen sulfide, sulfamate and trithiocarbonate were effective PgiCA inhibitors with inhibition constants in the range of 41-97 ?M. Other effective inhibitors were diethyldithiocarbamate, sulfamide, and phenylboronic acid, with KIs of 4.0-9.8 ?M. The role of this enzyme as a possible virulence factor of P. gingivalis is poorly understood at the moment but its good catalytic activity and the possibility to be inhibited by a large number of compounds may lead to interesting developments in the field.

Del Prete S; Vullo D; De Luca V; Carginale V; Scozzafava A; Supuran CT; Capasso C

2013-07-01

191

A highly catalytically active ?-carbonic anhydrase from the pathogenic anaerobe Porphyromonas gingivalis and its inhibition profile with anions and small molecules.  

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Carbonic anhydrases (CAs, EC 4.2.1.1) belonging to the ?-class are present in archaea, bacteria and plants but, except the Methanosarcina thermophila enzymes CAM and CAMH, they were poorly characterized so far. Here we report a new such enzyme (PgiCA), the ?-CA from the oral cavity pathogenic bacterium Porphyromonas gingivalis, the main causative agent of periodontitis. PgiCA showed a good catalytic activity for the CO2 hydration reaction, comparable to that of the human (h) isoform hCA I. Inorganic anions such as thiocyanate, cyanide, azide, hydrogen sulfide, sulfamate and trithiocarbonate were effective PgiCA inhibitors with inhibition constants in the range of 41-97 ?M. Other effective inhibitors were diethyldithiocarbamate, sulfamide, and phenylboronic acid, with KIs of 4.0-9.8 ?M. The role of this enzyme as a possible virulence factor of P. gingivalis is poorly understood at the moment but its good catalytic activity and the possibility to be inhibited by a large number of compounds may lead to interesting developments in the field. PMID:23769640

Del Prete, Sonia; Vullo, Daniela; De Luca, Viviana; Carginale, Vincenzo; Scozzafava, Andrea; Supuran, Claudiu T; Capasso, Clemente

2013-05-28

192

Genotipificación de los genes rgpA y kgp que codifican para las gingipaínas de Porphyromonas gingivalis/ Genotyping of rgpA and kgp genes encoding Pophyromonas gingivalis gingipains  

Scientific Electronic Library Online (English)

Full Text Available Abstract in spanish Porphyromonas gingivalis es un microorganismo fuertemente asociado con la etiología de la periodontitis. Esta bacteria posee varios factores de virulencia, dentro de los que destacan las gingipaínas, debido a sus múltiples acciones relacionadas con la destrucción de la matriz extracelular del tejido conectivo periodontal, la modulación del sistema inmune del hospedero y la estimulación de la expresión de citoquinas pro-inflamatorias. Estas proteinasas tienen afinid (more) ades específicas siendo Arg-gingipaínas (RgpA y RgpB, codificadas por los genes rgpA y rgpB, respectivamente) y Lys-gingipaínas (Kgp, codificada por el gen kgp). Se ha descrito que existen polimorfismos en los genes que codifican para esta proteinasas. El objetivo del presente estudio fue describir la frecuencia de los genotipos identificados para los genes rgpA y kgp en aislados clínicos de P. gingivalis, obtenidos desde pacientes con periodontitis. Para ello se utilizó amplificación por PCR de los genes rgpA y kgp, seguido de análisis de restricción. De un total de 47 aislados provenientes de 4 individuos con periodontitis crónica y 2 con periodontitis agresiva, se genotipificaron 38 aislados para el gen rgpA, exhibiendo la totalidad de éstos el patrón electroforético A (100%). Para el gen kgp se genotipificaron 43 aislados, presentando 28 de ellos (65.2%) el perfil electroforético kgp-I y 15 aislados (34.8%) el perfil kgp-II. En los aislados provenientes de un individuo fue posible apreciar ambos genotipos descritos para el gen kgp. Los resultados indican un predominio del patrón electroforético A (rgpA) y que el genotipo kgp-I fue el más frecuentemente encontrado de los genotipos kgp. Abstract in english Porphyromonas gingivalis is a microorganism strongly associated with the etiology of periodontitis. This periodontal bacterium possesses an array of virulence factors, among which gingipains have a key importance, being involved with extracellular matrix destruction of periodontal tissues, modulation of host immune response and stimulation in the production of pro-inflammatory cytokines by different types of cells. These proteinases have specific affinities, being Arg-gin (more) gipains (RgpA and RgpB, encoded by rgpA and rgpB genes, respectively) and Lys-gingipains (Kgp, encoded by the kgp gene). It has been described that there are polymorphisms in the genes encoding for gingipains. Therefore, the aim of the present study was to describe the frequency of rgpA and kgp genotypes in clinical isolates of P. gingivalis obtained from periodontitis patients. For determining the rgpA and kgp genotypes, we used PCR amplification and restriction analysis. From 47 isolates obtained from 4 individuals with chronic periodontitis and 2 subjects with aggressive periodontitis, 38 were typified for rgpA gene and all exhibited the electrophoretic pattern A (100%). For kgp gene, we characterized 43 isolates, 28 of them (65.2%) with the kgp-I electrophoretic profile and 15 isolates (34.8%) with the kgp-II profile. In the isolates belonging to one individual, we found both genotypes of kgp gene. The results indicate a clear predominance of the electrophoretic pattern A (for rgpA gene) and kgp-I genotype was the most frequently found of the kgp genotypes.

Abusleme, L; Blanc, V; Léon, R; Gamonal, J; Silva, N

2012-12-01

193

Porphyromonas gingivalis, Treponema denticola and toll-like receptor 2 are associated with hypertensive disorders in placental tissue: a case-control study.  

UK PubMed Central (United Kingdom)

AIM(S): To explore the associations between the presence of periodontal pathogens and the expression of toll-like receptors (TLR-2 and TLR-4) in the placental tissue of patients with hypertensive disorders compared to the placentas of healthy normotensive patients. MATERIAL AND METHODS: A case-control study was performed. From a cohort composed of 126 pregnant women, 33 normotensive healthy pregnant women were randomly selected, and 25 cases of patients with hypertensive disorders of pregnancy, including gestational hypertension and pre-eclampsia, were selected. Placental biopsy was obtained after aseptic placental collection at the time of delivery. All of the samples were processed and analysed for the detection of Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Fusobacterium nucleatum, Treponema denticola and Tannerella forsythia using the polymerase chain reaction (PCR) technique. Determination of the expressions of TLR-2 and TLR-4 was performed in samples of total purified protein isolated from placental tissues and analysed by ELISA. The data were assessed using descriptive statistics. The associations among variables were estimated through multiple logistic regression models and the Mann-Whitney test to evaluate the differences between the two groups. RESULTS: A significant increase was observed in the expression of TLR-2 in the placentas of patients with hypertensive disorders (p = 0.04). Additionally, the multiple logistic regression models demonstrated an association between the presence of T. denticola and P. gingivalis in placental tissues and hypertensive disorders (OR: 9.39, p = 0.001, CI 95% 2.39-36.88 and OR: 7.59, p = 0.019, CI 95% 1.39-41.51, respectively). CONCLUSIONS: In the present study, pregnant women with periodontal disease presented an association in the placental tissue between the presence of T. denticola and P. gingivalis and hypertensive disorders. Additionally, increased expression of TLR-2 was observed. However, further studies are required to determine the specific roles of periodontal pathogens and TLRs in the placental tissue of patients with pregnancy-related hypertensive disorders.

Chaparro A; Blanlot C; Ramírez V; Sanz A; Quintero A; Inostroza C; Bittner M; Navarro M; Illanes SE

2013-05-01

194

CCL3 and CXCL12 production in vitro by dental pulp fibroblasts from permanent and deciduous teeth stimulated by Porphyromonas gingivalis LPS  

Scientific Electronic Library Online (English)

Full Text Available Abstract in english Objective: The aim of this study was to compare the production of the chemokines CCL3 and CXCL12 by cultured dental pulp fibroblasts from permanent (PDPF) and deciduous (DDPF) teeth under stimulation by Porphyromonas gingivalis LPS (PgLPS). Material and Methods: Primary culture of fibroblasts from permanent (n=3) and deciduous (n=2) teeth were established using an explant technique. After the fourth passage, fibroblasts were stimulated by in (more) creasing concentrations of PgLPS (0 – 10 µg/mL) at 1, 6 and 24 h. The cells were tested for viability through MTT assay, and production of the chemokines CCL3 and CXCL12 was determined through ELISA. Comparisons among samples were performed using One-way ANOVA for MTT assay and Two-way ANOVA for ELISA results. Results: Cell viability was not affected by the antigen after 24 h of stimulation. PgLPS induced the production of CCL3 by dental pulp fibroblasts at similar levels for both permanent and deciduous pulp fibroblasts. Production of CXCL12, however, was significantly higher for PDPF than DDPF at 1 and 6 h. PgLPS, in turn, downregulated the production of CXCL12 by PDPF but not by DDPF. Conclusion: These data suggest that dental pulp fibroblasts from permanent and deciduous teeth may present a differential behavior under PgLPS stimulation.

Sipert, Carla Renata; Morandini, Ana Carolina de Faria; Modena, Karin Cristina da Silva; Dionisio, Thiago Jose; Machado, Maria Aparecida Andrade Moreira; Oliveira, Sandra Helena Penha de; Campanelli, Ana Paula; Santos, Carlos Ferreira

2013-04-01

195

Identification of a gingipain-sensitive surface ligand of Porphyromonas gingivalis that induces Toll-like receptor 2- and 4-independent NF-kappaB activation in CHO cells.  

Science.gov (United States)

Porphyromonas gingivalis is a major periodontal pathogen that has the pathogenic proteinases Arg-specific gingipain and Lys-specific gingipain. We previously found that a cell surface component on P. gingivalis is able to induce Toll-like receptor 2 (TLR2)- and TLR4-independent signaling in 7.19 cells and that this component can be degraded by gingipains. In this study, we purified this component from the P. gingivalis gingipain-null mutant KDP136 and obtained two candidate proteins. Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry analysis showed that the proteins, with molecular masses of 123 and 43 kDa, were encoded by PGN_0748 and PGN_0728 (pgm6), respectively, in the P. gingivalis ATCC 33277 genome sequence. The PGN_0748-encoded protein, which we refer to as gingipain-sensitive ligand A (GslA), reacted with antiserum that could effectively inhibit the activity of KDP136 to induce NF-kappaB activation in 7.19 cells, but Pgm6 did not. To further determine what protein is responsible for the NF-kappaB activation, we constructed gslA, pgm6, and pgm6 pgm7 deletion mutants from KDP136. When 7.19 cells were exposed to those mutants, the gslA deletion mutant did not induce NF-kappaB activation, whereas the pgm6 and pgm6 pgm7 deletion mutants did. Furthermore, NF-kappaB activation in 7.19 cells induced by KDP136 was partially inhibited by antiserum against a recombinant protein expressed from the 5'-terminal third of gslA. These results indicate that GslA is one of the factors that induce NF-kappaB activation in 7.19 cells. Interestingly, the gslA gene was present in four of seven P. gingivalis strains tested. This restricted distribution might be associated with the virulence potential of each strain. PMID:19667049

Haruyama, Koki; Yoshimura, Atsutoshi; Naito, Mariko; Kishimoto, Mami; Shoji, Mikio; Abiko, Yoshimitsu; Hara, Yoshitaka; Nakayama, Koji

2009-08-10

196

Identification of a gingipain-sensitive surface ligand of Porphyromonas gingivalis that induces Toll-like receptor 2- and 4-independent NF-kappaB activation in CHO cells.  

UK PubMed Central (United Kingdom)

Porphyromonas gingivalis is a major periodontal pathogen that has the pathogenic proteinases Arg-specific gingipain and Lys-specific gingipain. We previously found that a cell surface component on P. gingivalis is able to induce Toll-like receptor 2 (TLR2)- and TLR4-independent signaling in 7.19 cells and that this component can be degraded by gingipains. In this study, we purified this component from the P. gingivalis gingipain-null mutant KDP136 and obtained two candidate proteins. Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry analysis showed that the proteins, with molecular masses of 123 and 43 kDa, were encoded by PGN_0748 and PGN_0728 (pgm6), respectively, in the P. gingivalis ATCC 33277 genome sequence. The PGN_0748-encoded protein, which we refer to as gingipain-sensitive ligand A (GslA), reacted with antiserum that could effectively inhibit the activity of KDP136 to induce NF-kappaB activation in 7.19 cells, but Pgm6 did not. To further determine what protein is responsible for the NF-kappaB activation, we constructed gslA, pgm6, and pgm6 pgm7 deletion mutants from KDP136. When 7.19 cells were exposed to those mutants, the gslA deletion mutant did not induce NF-kappaB activation, whereas the pgm6 and pgm6 pgm7 deletion mutants did. Furthermore, NF-kappaB activation in 7.19 cells induced by KDP136 was partially inhibited by antiserum against a recombinant protein expressed from the 5'-terminal third of gslA. These results indicate that GslA is one of the factors that induce NF-kappaB activation in 7.19 cells. Interestingly, the gslA gene was present in four of seven P. gingivalis strains tested. This restricted distribution might be associated with the virulence potential of each strain.

Haruyama K; Yoshimura A; Naito M; Kishimoto M; Shoji M; Abiko Y; Hara Y; Nakayama K

2009-10-01

197

The Porphyromonas gingivalis HmuY haemophore binds gallium(iii), zinc(ii), cobalt(iii), manganese(iii), nickel(ii), and copper(ii) protoporphyrin IX but in a manner different to iron(iii) protoporphyrin IX.  

UK PubMed Central (United Kingdom)

Porphyromonas gingivalis, a major etiological agent of chronic periodontitis, acquires haem from host haemoproteins through a haem transporter HmuR and a haemophore HmuY. The aim of this study was to analyse the binding specificity of HmuY towards non-iron metalloporphyrins which may be employed as antimicrobials to treat periodontitis. HmuY binds gallium(iii), zinc(ii), cobalt(iii), manganese(iii), nickel(ii), and copper(ii) protoporphyrin IX but in a manner different to iron(iii) protoporphyrin IX which uses His(134) and His(166) as axial ligands. The metal ions in Ga(iii)PPIX and Zn(ii)PPIX can accept only His(166) as an axial ligand, whereas nickel(ii) and copper(ii) interact exclusively with His(134). Two forms of pentacoordinate manganese(iii) are present in the Mn(iii)PPIX-HmuY complex since the metal accepts either His(134) or His(166) as a single axial ligand. The cobalt ion is hexacoordinate in the Co(iii)PPIX-HmuY complex and binds His(134) and His(166) as axial ligands; however, some differences in their environments exist. Despite different coordination modes of the central metal ion, gallium(iii), zinc(ii), cobalt(iii), and manganese(iii) protoporphyrin IX bound to the HmuY haemophore cannot be displaced by excess haem. All of the metalloporphyrins examined bind to a P. gingivalis wild-type strain with higher ability compared to a mutant strain lacking a functional hmuY gene, thus corroborating binding of non-iron metalloporphyrins to purified HmuY protein. Our results further clarify the basis of metalloporphyrin acquisition by P. gingivalis and add to understanding of the interactions with porphyrin derivatives which exhibit antimicrobial activity against P. gingivalis.

Wójtowicz H; Bielecki M; Wojaczy?ski J; Olczak M; Smalley JW; Olczak T

2013-04-01

198

Lipopolysaccharide affinity for titanium implant biomaterials.  

UK PubMed Central (United Kingdom)

STATEMENT OF PROBLEM: Lipopolysaccharide (LPS) affinity for titanium implant biomaterials could affect crevicular LPS concentrations and thereby influence periimplant inflammation. PURPOSE OF STUDY: The purpose of this study was to evaluate Porphyromonas gingivalis and Escherichia coli LPS affinity for titanium biomaterials groups that differed in surface oxide composition and surface roughness. MATERIAL AND METHOD: Polished and abraded grade 1 commercially pure titanium and grade 5 alloyed extra low interstitial titanium specimens were treated with 10 EU/mm2 and radiolabeled LPS. RESULTS: The resultant mean +/- SD LPS adherence values ranged from 4.17 +/- 0.29 to 4.79 +/- 0.40 EU/ mm2. No difference in adherence and elution was indicated on the basis of LPS type, surface oxide composition, or surface roughness. Moreover, P. gingivalis and F. coli LPS desorption was below detection. CONCLUSION: Clinically, the high affinity of both LPS types for titanium biomaterials may adversely influence the periimplant tissue response.

Nelson SK; Knoernschild KL; Robinson FG; Schuster GS

1997-01-01

199

Cyclooxygenase-2 S-nitrosylation in salivary gland acinar cell inflammatory responses to Porphyromonas gingivalis: modulatory effect of ghrelin  

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Full Text Available Disturbances in nitric oxide synthase (NOS) system and the excessive prostaglandin (PGE2) generation are well-recognized features of oral mucosal inflammatory responses to periodontopathic bacterium, P. gingivalis. Employing rat sublingual gland acinar cells, we show that P. gingivalis LPS-induced up-regulation in PGE2 generation and the enhancement in inducible (i) iNOS activity was associated with COX-2 activation through S-nitrosylation, and accompanied by the suppression in cSrc activity and the impairment in constitutive (c) cNOS phosphorylation. Further, we demonstrate that the countering effect of peptide hormone, ghrelin, on the LPS-induced changes was reflected in the increased cNOS activation through phosphorylation, repression in iNOS induction, and the reduction in PGE2 generation associated with the loss of COX-2 protein S-nitrosylation. Moreover, the effect of ghrelin on cNOS phosphorylation and the LPS-induced COX-2 S-nitrosylation was susceptible to the blockage by cSrc inhibition. Our findings suggest that P. gingivalis-induced up-regulation in iNOS leads to COX-2 S-nitrosylation and up-regulation in PGE2 generation, and that the countering effect of ghrelin is mediated through Src-dependent cNOS activation that is obligatory for the maintenance of iNOS gene suppression.

Bronislaw L. Slomiany; Amalia Slomiany

2011-01-01

200

[Effect of NF-?B on the expression of interleukin-6 induced by lipopolysaccharides of Porphyromonas endodontalis in MC3T3-E1 cells.  

UK PubMed Central (United Kingdom)

PURPOSE: To investigate the effect of NF-?B signaling on the expression of interleukin-6(IL-6) induced by lipopolysaccharides(LPS) extracted from Porphyromonas endodontalis(P.e) in MC3T3-El cells. METHODS: MC3T3-E1 cells were pretreated with BAY-117082 for 1 h, and then were treated with 10 mg/L P.e-LPS for different times. The translocation of NF-?B was observed by immunofluorescence. The expression of IL-6 was detected by reverse transcription polymerse chain reaction (RT-PCR) and enzyme-linked immuno sorbent assay (ELISA). Statistical analysis was performed using multi-way ANOVA and Dunnett's t test with SPSS 13.0 software package. RESULTS: The staining of NF-?B was mostly in cytoplasm in untreated cells. Rapid translocation of NF-?B into nucleus was observed in the cells stimulated for 30 min and mostly relocalization of NF-?B from nucleus to cytoplasm was observed after 60 min. Pretreatment with 10 ?mol/L BAY-117082 for 1h significantly inhibited P.e-LPS-induced translocation of NF-?B .The mRNA and proteins of IL-6 decreased significantly after pretreatment with 10 ?mol/L BAY-117082 and the expression of IL-6 proteins was reduced from (774.983±6.585) ng/L to (377.384±14.620) ng/L (P<0.01). The group of treatment with BAY-117082 alone had no significant difference from the blank control group. CONCLUSIONS: P.e-LPS can induce translocation of NF-?B in mouse osteoblast MC3T3-El, and P.e-LPS may induce the expression of IL-6 in mouse osteoblast through the signaling of NF-?B. Supported by Science and Technology Program of Liaoning Province(2011225020).

Yu YQ; Guo JJ; Qiu LH; Lv Y; Jia G; Guo Y

2013-08-01

 
 
 
 
201

Deletion of Lipoprotein PG0717 in Porphyromonas gingivalis W83 Reduces Gingipain Activity and Alters Trafficking in and Response by Host Cells  

Science.gov (United States)

P. gingivalis (Pg), a causative agent of chronic generalized periodontitis, has been implicated in promoting cardiovascular disease. Expression of lipoprotein gene PG0717 of Pg strain W83 was found to be transiently upregulated during invasion of human coronary artery endothelial cells (HCAEC), suggesting this protein may be involved in virulence. We characterized the virulence phenotype of a PG0717 deletion mutant of pg W83. There were no differences in the ability of W83?717 to adhere and invade HCAEC. However, the increased proportion of internalized W83 at 24 hours post-inoculation was not observed with W83?717. Deletion of PG0717 also impaired the ability of W83 to usurp the autophagic pathway in HCAEC and to induce autophagy in Saos-2 sarcoma cells. HCAEC infected with W83?717 also secreted significantly greater amounts of MCP-1, IL-8, IL-6, GM-CSF, and soluble ICAM-1, VCAM-1, and E-selectin when compared to W83. Further characterization of W83?717 revealed that neither capsule nor lipid A structure was affected by deletion of PG0717. Interestingly, the activity of both arginine (Rgp) and lysine (Kgp) gingipains was reduced in whole-cell extracts and culture supernatant of W83?717. RT-PCR revealed a corresponding decrease in transcription of rgpB but not rgpA or kgp. Quantitative proteome studies of the two strains revealed that both RgpA and RgpB, along with putative virulence factors peptidylarginine deiminase and Clp protease were significantly decreased in the W83?717. Our results suggest that PG0717 has pleiotropic effects on W83 that affect microbial induced manipulation of host responses important for microbial clearance and infection control.

Reyes, Leticia; Eiler-McManis, Eileen; Rodrigues, Paulo H.; Chadda, Amandeep S.; Wallet, Shannon M.; Belanger, Myriam; Barrett, Amanda G.; Alvarez, Sophie; Akin, Debra; Dunn, William A.; Progulske-Fox, Ann

2013-01-01

202

Isolation of a variant Porphyromonas sp. from polymicrobial infections in central bearded dragons (Pogona vitticeps).  

UK PubMed Central (United Kingdom)

Isolates of gram-negative anaerobic bacteria from reptiles have only occasionally been identified to the genus and species level in the veterinary medical literature. In particular, reports identifying Porphyromonas spp. from infections in reptiles are scarce. The present report describes unique Porphyromonas isolates obtained from necrosuppurative infections in central bearded dragons (Pogona vitticeps). The isolates grew in the presence of oxygen, were strongly hemolytic, and did not produce detectable black, iron porphyrin pigment. Biochemical identification kit numeric biocodes gave high but unreliable probabilities (>99.9%) for identification as Porphyromonas gingivalis. Partial 16S ribosomal RNA gene sequences of the isolates were identical to each other and shared 91% identity with those of Porphyromonas gulae. The isolates may represent a new reptile-associated Porphyromonas species.

Bemis DA; Greenacre CB; Bryant MJ; Jones RD; Kania SA

2011-01-01

203

Isolation of a variant Porphyromonas sp. from polymicrobial infections in central bearded dragons (Pogona vitticeps).  

Science.gov (United States)

Isolates of gram-negative anaerobic bacteria from reptiles have only occasionally been identified to the genus and species level in the veterinary medical literature. In particular, reports identifying Porphyromonas spp. from infections in reptiles are scarce. The present report describes unique Porphyromonas isolates obtained from necrosuppurative infections in central bearded dragons (Pogona vitticeps). The isolates grew in the presence of oxygen, were strongly hemolytic, and did not produce detectable black, iron porphyrin pigment. Biochemical identification kit numeric biocodes gave high but unreliable probabilities (>99.9%) for identification as Porphyromonas gingivalis. Partial 16S ribosomal RNA gene sequences of the isolates were identical to each other and shared 91% identity with those of Porphyromonas gulae. The isolates may represent a new reptile-associated Porphyromonas species. PMID:21217036

Bemis, David A; Greenacre, Cheryl B; Bryant, Mary Jean; Jones, Rebekah D; Kania, Stephen A

2011-01-01

204

[Effect of lipopolysaccharide on proliferation and inflammatory factors expression of human periodontal ligament stem cells].  

UK PubMed Central (United Kingdom)

OBJECTIVE: To investigate the effect of Porphyromonas gingivalis lipopolysaccharide (LPS) on proliferation and inflammatory factors expression of human periodontal ligament stem cells (HPDLSCs). METHODS: HPDLSCs were cultivated and identified. Experiment was divided into 3 groups according to culture solution: Group A with alpha-MEM culture solution containing 10 microg.mL-1 LPS, group B with supernatant fluid containing 10ng.mL-1 LPS stimulated monocyte, group C with alpha-MEM culture solution. The proliferation ability of HPDLSCs was analyzed by MTF assay. The expression levels of interleukin-1beta (IL-1beta), interleukin-6 (IL-6), tumor necrosis factor (TNF-alpha) mRNA of HPDLSCs were detected by reverse transcriptase polymerase chain reaction(RT-PCR). RESULTS: HPDLSCs had clonality, bone and fat differentiation ability. Compared with group C, the proliferation ability of HPDLSCs of group A and group B was significantly inhibited, and the proliferation ability of HPDLSCs of group B were more significantly inhibited than that of group A (P<0.05). The expression of IL-1beta, IL-6 and TNF-alpha mRNA of group A and group B increased compared with the control group, and the expression of IL-1beta, IL-6 and TNF-alpha mRNA of group B increased more than that of group A (P<0.05). CONCLUSION: Porphyromonas gingivalis may inhibit the proliferation of HPDLSCs directly or indirectly through LPS and increase expression of inflammatory factor, exacerbate periodontal inflammatory tissue damage and delay the self-repairing of periodontal tissue.

Wang L; Xia J; Liu Q; Jin Y

2013-06-01

205

Inhibitory effects of Jixueteng on P. gingivalis-induced bone loss and osteoclast differentiation.  

UK PubMed Central (United Kingdom)

OBJECTIVE: The aim of this study was to investigate the possibility of Jixueteng as a preventive and therapeutic drug for the periodontitis. We investigated the inhibitory effects of Jixueteng on Porphyromonas gingivalis-induced bone loss in mice, antibacterial activity against P. gingivalis and differentiation of osteoclast and viability of cells. MATERIALS AND METHODS: Fifty-four male, 4-week-old C57BL/6N mice, were randomly divided into the following three groups of 18 mice each; group A served as the P. gingivalis non-infected control (sham group), group B was infected orally with P. gingivalis and group C was administered Jixueteng extract in drinking water and was then infected with P. gingivalis. In order to evaluate the effect of Jixueteng, the distance from the alveolar bone crest to the cemento-enamel junction was determined. P. gingivalis suspension was exposed for 1, 15 and 60 min to 5 ml of the Jixueteng extract. Furthermore, to clarify the mechanism of the inhibitory effects of Jixueteng on osteoclast formation, Jixueteng extract was added to the culture of mouse bone marrow cells, osteoclast precursor. RESULTS: Administration of Jixueteng along with P. gingivalis infection significantly reduced alveolar bone loss compared to P. gingivalis infection. Jixueteng treatment at the concentration of 0.01% significantly inhibited osteoclast formation. The addition of Jixueteng extract (0.1%, 0.01%, and 0.001%) to the culture showed a significant inhibition of the number of surviving osteoclasts in a dose-dependent manner. CONCLUSION: Jixueteng has an antibacterial activity against P. gingivalis and inhibitory effects on osteoclastogenesis, it may be useful as a therapeutic drug in the treatment of P. gingivalis-induced periodontitis.

Toyama T; Todoki K; Takahashi Y; Watanabe K; Takahashi SS; Sugiyama S; Lee MC; Hamada N

2012-11-01

206

Porphyromonas gingivalis stimulates TACE production by T cells  

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INTRODUCTION: Tumour necrosis factor-alpha converting enzyme (TACE), also known as ADAM17, is a membrane-bound metalloprotease and disintegrin. It is produced by a number of host cells and is known to shed and release cell-bound cytokines, particularly members of the tumour necrosis factor family. T...

Bostanci, N; Reddi, D; Rangarajan, M; Curtis, M A; Belibasakis, G N

207

Human ?- and ?-Defensins Bind to Immobilized Adhesins from Porphyromonas gingivalis?  

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Human neutrophil peptide ?-defensins (HNPs) and human ?-defensins (HBDs) are small well-characterized peptides with broad antimicrobial activities and a diversity of innate immune functions. Although the interactions of defensins with bacteria and their membranes have been well characterized, the in...

Dietrich, Deborah E.; Xiao, Xiangjun; Dawson, Deborah V.; Bélanger, Myriam; Xie, Hua; Progulske-Fox, Ann; Brogden, Kim A.

208

Bacteroides gingivalis and Bacteroides intermedius recognize different sites on human fibrinogen  

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Bacteroides (Porphyromonas) gingivalis and Bacteroides (Porphyromonas) intermedius have been implicated in the etiology of human periodontal diseases. These organisms are able to bind and degrade human fibrinogen, and these interactions may play a role in the pathogenesis of periodontal disease. In attempts to map the bacterial binding sites along the fibrinogen molecule, we have found that strains of B. gingivalis and B. intermedius, respectively, recognize spatially distant and distinct sites on the fibrinogen molecule. Isolated reduced and alkylated alpha-, beta-, and gamma-fibrinogen chains inhibited binding of 125I-fibrinogen to both Bacteroides species in a concentration-dependent manner. Plasmin fragments D and to some extent fragment E, however, produced a concentration-dependent inhibition of 125I-fibrinogen binding to B. intermedius strains but did not affect binding of 125I-fibrinogen to B. gingivalis strains. Radiolabeled fibrinogen chains and fragments were compared with 125I-fibrinogen with respect to specificity and reversibility of binding to bacteria. According to these criteria, gamma chain most closely resembled the native fibrinogen molecule in behavior toward B. gingivalis strains and fragments D most closely resembled fibrinogen in behavior toward B. intermedius strains. The ability of anti-human fibrinogen immunoglobulin G (IgG) to inhibit binding of 125I-fibrinogen to B. intermedius strains was greatly reduced by absorbing the IgG with fragments D. Absorbing the IgG with fragments D had no effect on the ability of the antibody to inhibit binding of 125I-fibrinogen to B. gingivalis strains. A purified staphylococcal fibrinogen-binding protein blocked binding of 125I-fibrinogen to B. intermedius strains but not to B. gingivalis strains.

1990-01-01

209

Bacteroides gingivalis and Bacteroides intermedius recognize different sites on human fibrinogen  

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Bacteroides (Porphyromonas) gingivalis and Bacteroides (Porphyromonas) intermedius have been implicated in the etiology of human periodontal diseases. These organisms are able to bind and degrade human fibrinogen, and these interactions may play a role in the pathogenesis of periodontal disease. In attempts to map the bacterial binding sites along the fibrinogen molecule, we have found that strains of B. gingivalis and B. intermedius, respectively, recognize spatially distant and distinct sites on the fibrinogen molecule. Isolated reduced and alkylated alpha-, beta-, and gamma-fibrinogen chains inhibited binding of 125I-fibrinogen to both Bacteroides species in a concentration-dependent manner. Plasmin fragments D and to some extent fragment E, however, produced a concentration-dependent inhibition of 125I-fibrinogen binding to B. intermedius strains but did not affect binding of 125I-fibrinogen to B. gingivalis strains. Radiolabeled fibrinogen chains and fragments were compared with 125I-fibrinogen with respect to specificity and reversibility of binding to bacteria. According to these criteria, gamma chain most closely resembled the native fibrinogen molecule in behavior toward B. gingivalis strains and fragments D most closely resembled fibrinogen in behavior toward B. intermedius strains. The ability of anti-human fibrinogen immunoglobulin G (IgG) to inhibit binding of 125I-fibrinogen to B. intermedius strains was greatly reduced by absorbing the IgG with fragments D. Absorbing the IgG with fragments D had no effect on the ability of the antibody to inhibit binding of 125I-fibrinogen to B. gingivalis strains. A purified staphylococcal fibrinogen-binding protein blocked binding of 125I-fibrinogen to B. intermedius strains but not to B. gingivalis strains.

Lantz, M.S.; Allen, R.D.; Bounelis, P.; Switalski, L.M.; Hook, M. (Univ. of Alabama, Birmingham (USA))

1990-02-01

210

Regulation of ICAM-1 expression in gingival fibroblasts infected with high-glucose-treated P.?gingivalis.  

Science.gov (United States)

Porphyromonas gingivalis is a major pathogen in the initiation and progression of periodontal disease, which is recognized as a common complication of diabetes. ICAM-1 expression by human gingival fibroblasts (HGFs) is crucial for regulating local inflammatory responses in inflamed periodontal tissues. However, the effect of P.?gingivalis in a high-glucose situation in regulating HGF function is not understood. The P.?gingivalis strain CCUG25226 was used to study the mechanisms underlying the modulation of HGF ICAM-1 expression by invasion of high-glucose-treated P.?gingivalis (HGPg). A high-glucose condition upregulated fimA?mRNA expression in P.?gingivalis and increased its invasion ability in HGFs. HGF invasion with HGPg induced increases in the expression of ICAM-1. By using specific inhibitors and short hairpin RNA (shRNA), we have demonstrated that the activation of p38 MAPK and Akt pathways is critical for HGPg-induced ICAM-1 expression. Luciferase reporters and chromatin immunoprecipitation assays suggest that HGPg invasion increases NF-?B- and Sp1-DNA-binding activities in HGFs. Inhibition of NF-?B and Sp1 activations blocked the HGPg-induced ICAM-1 promoter activity and expression. The effect of HGPg on HGF signalling and ICAM-1 expression is mediated by CXC chemokine receptor 4 (CXCR4). Our findings identify the molecular pathways underlying HGPg-dependent ICAM-1 expression in HGFs, providing insight into the effect of P.?gingivalis invasion in HGFs. PMID:23551616

Chang, Li-Ching; Kuo, Hsing-Chun; Chang, Shun-Fu; Chen, Heng Jung; Lee, Kam-Fai; Lin, Tseng-Hsi; Huang, Ting-Ying; Choe, Chu-Shan; Lin, Li-Tsen; Chen, Cheng-Nan

2013-04-29

211

Oxidized galectin-1 reduces lipopolysaccharide-induced increase of proinflammatory cytokine mRNA in cultured macrophages.  

UK PubMed Central (United Kingdom)

BACKGROUND: Periodontitis is prevalent in older humans. Limiting the inflammation associated with periodontitis may provide a therapy for this condition, because Gram-negative bacteria expressing lipopolysaccharide (LPS) have a key role in initiation of inflammation by activating macrophage functions. Because oxidized galectin-1 regulates macrophage functions in other systems, we sought to establish whether this galectin-1 mRNA is expressed in the oral cavity, and whether it could dampen LPS-induced macrophage activation in vitro. METHODS: Using the reverse transcriptase polymerase chain reaction (RT-PCR), we measured galectin-1 mRNA expression to clarify its localization to rat gingival tissues and studied the effect of Porphyromonas gingivalis challenge on galectin-1 expression. Next, we tested the effects of adding oxidized galectin-1 to cultured LPS-activated peritoneal macrophages on mRNA expression of proinflammatory factors by RT-PCR and real-time RT-PCR. RESULTS: We established that galectin-1 mRNA is expressed in gingival tissues and also showed that galectin-1 mRNA was significantly increased by challenge with P. gingivalis, indicating that galectin-1 may regulate oral inflammation. On the other hand, LPS 100 ng/mL in serum-containing medium induced macrophages to upregulate mRNA associated with a proinflammatory response, ie, interleukins 1? and 6, and inducible nitric oxide synthase. We showed that application of 0.1-10 ng/mL of oxidized galectin-1 to LPS-treated macrophages reduced the intense LPS- induced increase by serum in proinflammatory mRNA expression in a concentration-dependent manner. Furthermore, application of oxidized galectin-1 10 ng/mL to LPS-treated macrophages in serum-free medium also showed a similar effect on LPS activity. CONCLUSION: Oxidized galectin-1 restricts the proinflammatory actions of LPS, and this protein could limit the negative effects of inflammation.

Kogawa Y; Nakajima K; Sasaguri K; Hamada N; Kawasaki H; Sato S; Kadoya T; Horie H

2011-01-01

212

ICAM-1 is Regulated by Rorphyromonas Gingivalis Through NOD1 and NOD2 Molecules in Periodontal Fibroblasts.  

UK PubMed Central (United Kingdom)

[Background]: The mechanism by which Porphyromonas gingivalis (P. gingivalis) regulates ICAM-1 expression in human periodontal ligament cells (hPDLCs) and human gingival fibroblasts (hGFs) is unknown. The aim of this study was to investigate whether nucleotide binding oligomerization domain-containing protein (NOD) 1 and NOD2 were involved in this process, and the clinical significance of ICAM-1 in periodontitis. [Methods]: hPDLCs and hGFs were treated with P. gingivalis, Tri-DAP (an agonist for NOD1) and MDP (an agonist for NOD2). Alternatively, cells transfected with siRNA targeting NOD1and NOD2 were treated with P. gingivalis. ICAM-1, NOD1 and NOD2 were detected at mRNA and protein levels. In addition, clinical examination were performed in 30 healthy controls and 40 patients with chronic periodontitis pre- and post-treatment, and serum soluble ICAM-1 (sICAM-1) levels in these individuals were detected by ELISA. [Results]: This study showed P. gingivalis caused an increase in ICAM-1, NOD1and NOD2 expression in periodontal fibroblasts. There was a linear correlation between ICAM-1 and NOD1/2 levels. Activation of NOD1/2 by the specific agonist led to the up-regulation of ICAM-1, whereas knocking down NOD1/2 caused a reduction in P. gingivalis-induced ICAM-1 production. Furthermore, sICAM-1 levels were higher in patients with chronic periodontitis than in healthy controls, and were positively related to the clinical periodontal parameters. After periodontal treatment, sICAM-1 levels decreased significantly. [Conclusion]: Our results indicate that sICAM-1 levels are correlated to the severity of periodontitis. NOD1/2 mediates P. gingivalis-induced ICAM-1 production in periodontal fibroblasts. NOD1/2 could be considered as potential targets for periodontal therapy.

Liu J; Duan J; Wang Y; Ouyang X

2013-05-01

213

Oxidized galectin-1 reduces lipopolysaccharide-induced increase of proinflammatory cytokine mRNA in cultured macrophages  

Directory of Open Access Journals (Sweden)

Full Text Available Yukie Kogawa1, Kou Nakajima1, Kenichi Sasaguri1, Nobushiro Hamada2, Haruhisa Kawasaki3, Sadao Sato1, Toshihiko Kadoya4, Hidenori Horie51Department of Orthodontics, 2Department of Oral Microbiology, Kanagawa Dental College, Yokosuka; 3Keio University, Kanagawa; 4Maebashi Institute of Technology, Maebashi; 5Research Center of Brain and Oral Science, Kanagawa Dental College, Yokosuka, JapanBackground: Periodontitis is prevalent in older humans. Limiting the inflammation associated with periodontitis may provide a therapy for this condition, because Gram-negative bacteria expressing lipopolysaccharide (LPS) have a key role in initiation of inflammation by activating macrophage functions. Because oxidized galectin-1 regulates macrophage functions in other systems, we sought to establish whether this galectin-1 mRNA is expressed in the oral cavity, and whether it could dampen LPS-induced macrophage activation in vitro.Methods: Using the reverse transcriptase polymerase chain reaction (RT-PCR), we measured galectin-1 mRNA expression to clarify its localization to rat gingival tissues and studied the effect of Porphyromonas gingivalis challenge on galectin-1 expression. Next, we tested the effects of adding oxidized galectin-1 to cultured LPS-activated peritoneal macrophages on mRNA expression of proinflammatory factors by RT-PCR and real-time RT-PCR.Results: We established that galectin-1 mRNA is expressed in gingival tissues and also showed that galectin-1 mRNA was significantly increased by challenge with P. gingivalis, indicating that galectin-1 may regulate oral inflammation. On the other hand, LPS 100 ng/mL in serum-containing medium induced macrophages to upregulate mRNA associated with a proinflammatory response, ie, interleukins 1? and 6, and inducible nitric oxide synthase. We showed that application of 0.1–10 ng/mL of oxidized galectin-1 to LPS-treated macrophages reduced the intense LPS-induced increase by serum in proinflammatory mRNA expression in a concentration-dependent manner. Furthermore, application of oxidized galectin-1 10 ng/mL to LPS-treated macrophages in serum-free medium also showed a similar effect on LPS activity.Conclusion: Oxidized galectin-1 restricts the proinflammatory actions of LPS, and this protein could limit the negative effects of inflammation.Keywords: periodontitis, inflammation, macrophage, lipopolysaccharide, galectin-1, proinflammatory factors

Yukie Kogawa; Kou Nakajima; Kenichi Sasaguri; et al

2011-01-01

214

Macrolide antibiotics like azithromycin increase lipopolysaccharide-induced IL-8 production by human gingival fibroblasts  

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Full Text Available Abstract Objective Macrolide antibiotics are reported to modulate the production of cytokines in various type of cells. We examined the effect of macrolide antibiotics on inflammatory cytokines (IL-6 and IL-8) and chemical mediator (PGE2) and also matrix metalloproteinases (MMPs) productions by human gingival fibroblasts (HGFs) treated with lipopolysaccharide (LPS). Methods The effect of macrolide antibiotics [erythromycin (EM), azithromycin (AZM) and josamycin (JOM)] on HGFs proliferation were examined by MTT assay. HGFs were treated with LPS from Porphyromonas gingivalis (PgLPS) and macrolide antibiotics, and IL-6, IL-8 and PGE2 levels were evaluated by ELISA. MMPs were detected by gelatin zymography. Results AZM slightly but significantly decreased HGFs proliferation, while EM and JOM did not affected. AZM increased PgLPS-induced IL-8 production dose-dependently, while AZM did not alter IL-6 and PGE2 productions. EM and JOM did not altered PgLPS-induced IL-6, IL-8 and PGE2 productions. All macrolide antibiotics did not alter MMPs production. These results indicate that macrolide antibiotics have no direct anti-inflammatory effect. However, the use of the inhibitors of cell signaling pathway failed to reveal the mechanism that AZM enhanced PgLPS-induced IL-8 production. Conclusion These results suggest macrolide antibiotics have an indirect anti-inflammatory effect as a result of their antimicrobial properties. Because AZM increased LPS-induced IL-8 production by HGFs, the possibility is considered that neutrophils may be migrated to periodontal tissue and phagocytize the periodontopathic bacteria more efficiently.

Kamemoto A; Ara T; Hattori T; Fujinami Y; Imamura Y; Wang P-L

2009-01-01

215

Specificity of antimicrobial peptide LL-37 to neutralize periodontopathogenic lipopolysaccharide activity in human oral fibroblasts.  

UK PubMed Central (United Kingdom)

BACKGROUND: The antimicrobial peptide LL-37 is known to have a potent lipopolysaccharide (LPS)-neutralizing activity in various cell types. Because of observed heterogeneity within periodontopathogenic LPS, the authors hypothesized that LL-37 had specificity to neutralize such LPS activity. The present study, therefore, aims to investigate the LPS-neutralizing activity of LL-37 to various periodontopathogenic LPS in interleukin-8 (IL-8) production after challenging them in human oral fibroblasts. METHODS: Human periodontal ligament fibroblasts (PDLF) and gingival fibroblasts (GF) were cultured from biopsies of periodontal ligament and gingival tissues. After cell confluence in 24-well plates, LPS (10 ?g/mL) from Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, and Aggregatibacter actinomycetemcomitans were added with or without LL-37 (10 ?g/mL). After 18 hours, the supernatant was collected and analyzed in IL-8 production by enzyme-linked immunosorbent assay. RESULTS: All periodontopathogenic LPS statistically significantly induced IL-8 production in both PDLF and GF (P <0.01). After neutralization with LL-37, both PDLF and GF showed a statistically significant reduction in IL-8 production compared with LPS-treated groups without LL-37 (P <0.01), and the percentage of reduction in IL-8 production in PDLF appeared to be higher than in GF. In addition, the percentage of reduction in IL-8 production varied considerably according to each periodontopathogenic LPS. CONCLUSIONS: The antimicrobial peptide LL-37 had an ability to suppress periodontopathogenic LPS-induced IL-8 production in both PDLF and GF. Its LPS-neutralizing activity revealed specificity to periodontopathogenic LPS and seemed to be dependent on the heterogeneity within LPS between different genera.

Suphasiriroj W; Mikami M; Shimomura H; Sato S

2013-02-01

216

The ability of the BANA test to detect different levels of P. gingivalis, T. denticola and T. forsythia  

Scientific Electronic Library Online (English)

Full Text Available Abstract in english The aim of this study was to evaluate the ability of the BANA Test to detect different levels of Porphyromonas gingivalis, Treponema denticola and Tannerella forsythia or their combinations in subgingival samples at the initial diagnosis and after periodontal therapy. Periodontal sites with probing depths between 5-7 mm and clinical attachment level between 5-10 mm, from 53 subjects with chronic periodontitis, were sampled in four periods: initial diagnosis (T0), immediat (more) ely (T1), 45 (T2) and 60 days (T3) after scaling and root planing. BANA Test and Checkerboard DNA-DNA hybridization identified red complex species in the subgingival biofilm. In all experimental periods, the highest frequencies of score 2 (Checkerboard DNA-DNA hybridization) for P. gingivalis, T. denticola and T. forsythia were observed when strong enzymatic activity (BANA) was present (p

Andrade, José Alexandre de; Feres, Magda; Figueiredo, Luciene Cristina de; Salvador, Sérgio Luiz; Cortelli, Sheila Cavalca

2010-06-01

217

Anti-Porphyromonas gingivalis and Anti-Inflammatory Activities of A-Type Cranberry Proanthocyanidins ?  

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A-type cranberry proanthocyanidins (AC-PACs) have recently been reported to be beneficial for human health, especially urinary tract health. The effect of these proanthocyanidins on periodontitis, a destructive disease of tooth-supporting tissues, needs to be investigated. The purpose of this study ...

La, Vu Dang; Howell, Amy B.; Grenier, Daniel

218

The ability of the BANA test to detect different levels of P. gingivalis, T. denticola and T. forsythia  

Directory of Open Access Journals (Sweden)

Full Text Available The aim of this study was to evaluate the ability of the BANA Test to detect different levels of Porphyromonas gingivalis, Treponema denticola and Tannerella forsythia or their combinations in subgingival samples at the initial diagnosis and after periodontal therapy. Periodontal sites with probing depths between 5-7 mm and clinical attachment level between 5-10 mm, from 53 subjects with chronic periodontitis, were sampled in four periods: initial diagnosis (T0), immediately (T1), 45 (T2) and 60 days (T3) after scaling and root planing. BANA Test and Checkerboard DNA-DNA hybridization identified red complex species in the subgingival biofilm. In all experimental periods, the highest frequencies of score 2 (Checkerboard DNA-DNA hybridization) for P. gingivalis, T. denticola and T. forsythia were observed when strong enzymatic activity (BANA) was present (p < 0.01). The best agreement was observed at initial diagnosis. The BANA Test sensitivity was 95.54% (T0), 65.18% (T1), 65.22% (T2) and 50.26% (T3). The specificity values were 12.24% (T0), 57.38% (T1), 46.27% (T2) and 53.48% (T3). The BANA Test is more effective for the detection of red complex pathogens when the bacterial levels are high, i.e. in the initial diagnosis of chronic periodontitis.

José Alexandre de Andrade; Magda Feres; Luciene Cristina de Figueiredo; Sérgio Luiz Salvador; Sheila Cavalca Cortelli

2010-01-01

219

Antigen Capture of Porphyromonas gingivalis by Human Macrophages Is Enhanced but Killing and Antigen Presentation Are Reduced by Endotoxin Tolerance?  

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The innate and the adaptive arms of the mucosal immune system must be coordinated to facilitate the control of pathogenic invasion while maintaining immune homeostasis. Toll-like receptors, able to activate the cell to produce bactericidal and inflammatory cytokines but also able to upregulate antig...

Muthukuru, Manoj; Cutler, Christopher W.

220

A molecular survey of S. mutans and P. gingivalis oral microbial burden in human saliva using Relative Endpoint Polymerase Chain Reaction (RE-PCR) within the population of a Nevada dental school revealed disparities among minorities  

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Full Text Available Abstract Background The University of Nevada, Las Vegas School of Dental Medicine recently opened an orthodontic treatment clinic to address the needs of the racially and ethnically diverse population of Southern Nevada, primarily focusing on the treatment and care of low-income and minority patients. Although orthodontic treatment and therapy has been shown to induce changes in the oral cavity, much of this evidence was collected from traditional White, teenage orthodontic clinic populations. The primary goal of this study was to describe the microbial burden of the cariogenic and periodontal pathogens, Streptococcus mutans and Porphyromonas gingivalis within the UNLV-SDM patient population. Methods Representative saliva samples were collected from healthy adult patients for DNA isolation. Relative endpoint polymerase chain reaction (RE-PCR) was performed to ascertain the presence and relative microbial burden of these oral pathogens. Results Nearly one quarter (13/56) or 23.3% of these patients had elevated levels of S. mutans, while (10/56) and 17.8% of these samples were found to have elevated levels of P. gingivalis, - with (90%) of P. gingivalis-positive samples from minority patients (X2?=?17.921, d.f. = 1; p? Conclusions These findings of elevated P. gingivalis levels, primarily among minority patients, may suggest underlying oral health practices contributing to adverse oral health conditions within this population. Oral health knowledge and practices among minority patients may be strongly influenced by other factors, including education and socioeconomic status, suggesting additional research may be needed to accurately determine the most appropriate standards for care and oral health education within this patient population.

Davis Jay; Freel Nicholas; Findley Allison; Tomlin Keaton; Howard Katherine M; Seran Clifford C; Cruz Patricia; Kingsley Karl

2012-01-01

 
 
 
 
221

Characterization of legionella lipopolysaccharide.  

UK PubMed Central (United Kingdom)

The lipopolysaccharide(LPS) of Legionella spp. is an immuno-dominant antigen and the basis for Legionella pneumophila serogroup classification. The LPS shows a peculiar structure composed of a very hydrophobic lipid A acylated by long chain fatty acids and an O-antigen-specific chain consisting of homopolymeric legionaminic acid. In this chapter we describe a method for the isolation of LPS from L. pneumophila. In the first part we describe the chemical purification, in the second part we outline the application of monoclonal antibody (mAb) in Western blot and immuno-localization by indirect immunofluorescence. This report does not describe physico-chemical methods that analyze the structure of lipopolysaccharide entities.

Lück C; Helbig JH

2013-01-01

222

Inflammatory cytokines are suppressed by light-emitting diode irradiation of P. gingivalis LPS-treated human gingival fibroblasts: inflammatory cytokine changes by LED irradiation.  

Science.gov (United States)

Human gingival fibroblasts (hGFs) play an important role in the inflammatory reaction to lipopolysaccharide (LPS) from P. gingivalis, which infects periodontal connective tissue. In addition, although light-emitting diode (LED) irradiation has been reported to have biostimulatory effects, including anti-inflammatory activity, the pathological mechanisms of these effects are unclear. This study examined the effects of 635-nm irradiation of P. gingivalis LPS-treated human gingival fibroblasts on inflammatory cytokine profiles and the mitogen-activated protein kinase (MAPK) pathway, which is involved in cytokine production. Gingival fibroblasts treated or not treated with P. gingivalis LPS were irradiated with 635-nm LED light, and cytokine profiles in the supernatant were assessed using a human inflammation antibody array. Expression of cyclooxyginase-2 (COX-2) protein and phosphorylation of extracellular signal-regulated kinase (ERK 1/2), p38, and c-Jun-N-terminal kinase (JNK) were assessed by Western-blot analysis to determine the effects on the MAPK pathway, and prostaglandin E(2) (PGE(2)) in the supernatant was measured using an enzyme-linked immunoassay. COX-2 protein expression and PGE(2) production were significantly increased in the LPS-treated group and decreased by LED irradiation. LPS treatment of gingival fibroblasts led to the increased release of the pro-inflammatory-related cytokines interleukin-6 (IL-6) and IL-8, whereas LED irradiation inhibited their release. Analysis of MAPK signal transduction revealed a considerable decrease in p38 phosphorylation in response to 635-nm radiation either in the presence or absence of LPS. In addition, 635-nm LED irradiation significantly promoted JNK phosphorylation in the presence of LPS. LED irradiation can inhibit activation of pro-inflammatory cytokines, mediate the MAPK signaling pathway, and may be clinically useful as an anti-inflammatory tool. PMID:21814735

Choi, HongRan; Lim, WonBong; Kim, InAe; Kim, JiSun; Ko, YoungJong; Kwon, Hyukil; Kim, SangWoo; Kabir, K M Ahsan; Li, Xiaojie; Kim, Oksu; Lee, YoungJoon; Kim, SeoYune; Kim, OkJoon

2011-08-04

223

Supported lipopolysaccharide bilayers.  

UK PubMed Central (United Kingdom)

In this report, the formation of supported lipopolysaccharide bilayers (LPS-SLBs) is studied with extracted native and glycoengineered LPS from Escherichia coli ( E. coli ) and Salmonella enterica sv typhimurium ( S. typhimurium ) to assemble a platform that allows measurement of LPS membrane structure and the detection of membrane tethered saccharide-protein interactions. We present quartz crystal microbalance with dissipation monitoring (QCM-D) and fluorescence recovery after photobleaching (FRAP) characterization of LPS-SLBs with different LPS species, having, for example, different molecular weights, that show successful formation of SLBs through vesicle fusion on SiO(2) surfaces with LPS fractions up to 50 wt %. The thickness of the LPS bilayers were investigated with AFM force-distance measurements which showed only a slight thickness increase compared to pure POPC SLBs. The E. coli LPS were chosen to study the saccharide-protein interaction between the Htype II glycan epitope and the Ralstonia solanacearum lectin (RSL). RSL specifically recognizes fucose sugars, which are present in the used Htype II glycan epitope and absent in the epitopes LPS1 and EY2. We show via fluorescence microscopy that the specific, but weak and multivalent interaction can be detected and discriminated on the LPS-SLB platform.

Kaufmann S; Ilg K; Mashaghi A; Textor M; Priem B; Aebi M; Reimhult E

2012-08-01

224

Supported lipopolysaccharide bilayers.  

Science.gov (United States)

In this report, the formation of supported lipopolysaccharide bilayers (LPS-SLBs) is studied with extracted native and glycoengineered LPS from Escherichia coli ( E. coli ) and Salmonella enterica sv typhimurium ( S. typhimurium ) to assemble a platform that allows measurement of LPS membrane structure and the detection of membrane tethered saccharide-protein interactions. We present quartz crystal microbalance with dissipation monitoring (QCM-D) and fluorescence recovery after photobleaching (FRAP) characterization of LPS-SLBs with different LPS species, having, for example, different molecular weights, that show successful formation of SLBs through vesicle fusion on SiO(2) surfaces with LPS fractions up to 50 wt %. The thickness of the LPS bilayers were investigated with AFM force-distance measurements which showed only a slight thickness increase compared to pure POPC SLBs. The E. coli LPS were chosen to study the saccharide-protein interaction between the Htype II glycan epitope and the Ralstonia solanacearum lectin (RSL). RSL specifically recognizes fucose sugars, which are present in the used Htype II glycan epitope and absent in the epitopes LPS1 and EY2. We show via fluorescence microscopy that the specific, but weak and multivalent interaction can be detected and discriminated on the LPS-SLB platform. PMID:22830310

Kaufmann, Stefan; Ilg, Karin; Mashaghi, Alireza; Textor, Marcus; Priem, Bernard; Aebi, Markus; Reimhult, Erik

2012-08-09

225

Lipopolysaccharide of Coxiella burnetii.  

UK PubMed Central (United Kingdom)

A lipopolysaccharide (LPS) is considered to be one of the major determinants of virulence expression and infection of virulent Coxiella burnetii. The LPSs from virulent phase I (LPS I) and from avirulent phase II (LPS II) bacteria were investigated for their chemical composition, structure and biological properties. LPS II is of rough (R) type in contrast to LPS I, which is phenotypically smooth (S) and contains a noticeable amount of two sugars virenose (Vir) and dihydrohydroxystreptose (Strep), which have not been found in other LPSs and can be considered as unique biomarkers of the bacterium. Both sugars were suggested to be located mostly in terminal positions of the O-specific chain of LPS I (O-PS I) and to be involved in the immunobiology of Q fever. There is a need to establish a more detailed chemical structure of LPS I in connection with prospective, deeper studies on mechanisms of pathogenesis and immunity of Q fever, its early and reliable diagnosis, and effective prophylaxis against the disease. This will also help to better understanding of host-pathogen interactions and contribute to improved modulation of pathological reactions which in turn are prerequisite for research and development of vaccines of new type. A fundamental understanding of C. burnetii LPS biosynthesis is still lacking. The intracellular nature of the bacterium, lack of genetic tools and its status as a selected agent have made elucidating basic physiological mechanisms challenging. The GDP-?-D-Vir biosynthetic pathway proposed most recently is an important initial step in this endeavour. The current advanced technologies providing the genetic tools necessary to screen C. burnetii mutants and propagate isogenic mutants might speed the discovery process.

Narasaki CT; Toman R

2012-01-01

226

[Lung cancer with bronchial stenosis due to foreign body and Entoameba gingivalis infection].  

Science.gov (United States)

Oral cavity infection by protozoarian agents may lead to pathologies such as stomatitis and gengivitis. An higher incidence has been reported in immunocompromised patients and in patients with dental disorders. Entoameba gingivalis localizes into oral cavity and in particular into interstitial and interdental spaces. Infection propagation to bronchial or lung parenchyma represents a complication. In this report the Authors, starting from a recently treated case, discuss on the incidence, complications and surgical management of lung infection by Entoameba gingivalis. PMID:21453594

Monaco, F; Mondello, B; Barone, M; Familiari, D; Sibilio, M; La Rocca, A; Lentini, S; Monaco, M

2011-03-01

227

Lipopolysaccharides and plant innate immunity.  

UK PubMed Central (United Kingdom)

Plants posses an innate immune system that has many parallels with those found in mammals and insects. A range of molecules of microbial origin called Microbe Associated Molecular Patterns (MAMPs) act to trigger basal defense responses in plants. These elicitors include lipopolysaccharides (LPS) from diverse Gram-negative bacteria. Both core oligosaccharide and the lipid A moieties of LPS as well as synthetic O-antigen oligosaccharides have activity in inducing defense responses in the model plant Arabidopsis thaliana. Very little is known of the mechanism of LPS perception by plants, although plant receptors for other MAMPs such as flagellin have been described. Recent work has implicated the Arabidopsis syntaxin PEN1 as a potential actor in LPS induction of plant defenses, which may suggest a role for vesicle trafficking in the signalling process.

Erbs G; Molinaro A; Dow JM; Newman MA

2010-01-01

228

Effect of environmental pH on enzyme activity and growth of Bacteroides gingivalis W50.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Since the pH of the gingival crevice increases from below neutrality in health to above pH 8 in disease, we decided to investigate the effect of environmental pH on the growth and enzyme activity of Bacteroides gingivalis W50. Cells were grown in a chemostat under hemin-excess conditions over a rang...

McDermid, A S; McKee, A S; Marsh, P D

229

[Monosaccharide composition of Ralstonia solanacearum lipopolysaccharides].  

Science.gov (United States)

Results of monosaccharide analysis of Ralstonia solanacearum lipopolysaccharides of the strains of different geographical origin are shown. Such monosaccharides as glucose, rhamnose and glucosamine prevail. Following the results of analysis, strains with supposedly truncated O-polysaccharide are revealed. As to their quantitative and monosaccharide composition of LPS the strains have demonstrated high level of heterogeneity. PMID:23516837

Hrytsa?, R V; Brovars'ka, O S; Varbanets', L D

230

[Monosaccharide composition of Ralstonia solanacearum lipopolysaccharides].  

UK PubMed Central (United Kingdom)

Results of monosaccharide analysis of Ralstonia solanacearum lipopolysaccharides of the strains of different geographical origin are shown. Such monosaccharides as glucose, rhamnose and glucosamine prevail. Following the results of analysis, strains with supposedly truncated O-polysaccharide are revealed. As to their quantitative and monosaccharide composition of LPS the strains have demonstrated high level of heterogeneity.

Hrytsa? RV; Brovars'ka OS; Varbanets' LD

2013-01-01

231

A rare case of Lemierre`s syndrome caused by Porphyromonas asaccharolytica.  

Science.gov (United States)

Lemierre's syndrome is only very rarely caused by Porphyromonas asaccharolytica. Here, we report the case of a 35-year-old man who developed a left peritonsillar abscess, thrombophlebitis of the left internal jugular vein, and septic embolization of both lungs. Anaerobic P. asaccharolytica was isolated in the blood cultures, and we subsequently confirmed the diagnosis as Lemierre's syndrome. Our case indicates that although P. asaccharolytica is not commonly found in oral cavities, this organism may still cause Lemierre's syndrome. Consequently, when it is detected in blood cultures, the treating physician should perform the medical examination while keeping in mind the possibility that the patient could have Lemierre's syndrome. PMID:23435719

Takeda, K; Kenzaka, T; Morita, Y; Kuroki, S; Kajii, E

2013-02-24

232

Equine meningo-encephalitis caused by Halicephalobus gingivalis: a case report observed during West Nile disease surveillance activities  

Directory of Open Access Journals (Sweden)

Full Text Available A seven-year-old horse was euthanised after exhibiting a severe and rapidly progressive neurological disorder. Tissue samples were despatched to the Italian Reference Centre for Animal Foreign Diseases (Istituto 'G. Caporale' in Teramo) for diagnosis. All laboratory tests for equine neurotropic viruses gave negative results. Scattered perivascular inflammatory infiltrates and several parasites that were morphologically classified as Halicephalobus gingivalis, were seen within the brain upon microscopic examination. Pathological findings led to the diagnosis of parasitic meningo-encephalitis caused by H. gingivalis. This case report confirms that halicephalobosis should be taken into account in the differential diagnosis of equine encephalopathy and it also highlights the value of a multidisciplinary approach to problem solving in veterinary medicine.

Gabriella Di Francesco; Giovanni Savini; Alessandro Maggi; Nicola Cavaliere; Anna Rita D’Angelo; Giuseppe Marruchella

2012-01-01

233

Identification of Legionella species by lipopolysaccharide antigen pattern.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Electrophoretic analysis of lipopolysaccharide (LPS) extracts from 430 previously serotyped Legionella isolates and 28 American Type Culture Collection (ATCC) non-Legionella pneumophila Legionella reference strains representing different Legionella species and serogroups has been performed. LPS was ...

Jürgens, D; Fehrenbach, F J

234

Complexing of bacterial lipopolysaccharide with lung surfactant.  

UK PubMed Central (United Kingdom)

Lipopolysaccharides (LPS) from Escherichia coli, Salmonella typhi, Klebsiella pneumoniae, Serratia marcescens, or Pseudomonas aeruginosa were mixed with pulmonary surfactant to investigate their in vitro interaction. After 6 h of incubation at 37 degrees C, LPS-surfactant mixtures were examined by sucrose density gradient centrifugation. The E. coli LPS-surfactant mixture was examined by immunoelectron microscopy with protein A-colloidal gold. The binding that occurred between LPS and the surfactant vesicles resulted in a complex with a density higher than the density of the surfactant alone. The protein A-colloidal gold identified LPS in the LPS-surfactant complexes. The toxicity of E. coli LPS was enhanced by complexing with the surfactant when compared with the intraperitoneal injection into CF1 mice, even at a 64:1 ratio of surfactant to LPS. The complexing of LPS and surfactant in the lung may alter the physiologic properties of surfactant that contribute to the physiopathological changes observed with some types of pneumonia.

Brogden KA; Cutlip RC; Lehmkuhl HD

1986-06-01

235

Composition of the lipopolysaccharide of Neisseria gonorrhoeae.  

UK PubMed Central (United Kingdom)

Analysis of glycose and fatty acid content of lipopolysaccharide extracted from 38 strains of Neisseria gonorrhoeae indicated that glycoses common to colonial types 1 to 5 were glucose, mannose, and galactose, N-acetylneuraminic acid, 2-keto-3-deoxyoctulosonic acid (KDO), glucosamine, and galactosamine were also invariably present. Virulent colonial types 1 and 2 contained no rhamnose, in contrast to avirulent types 3 to 5 and several strains of the nonpathogenic species N. sicca and N. lactamica. Fucose, characteristic of these nonpathogenic species, was not present in the gonococci. Variation in the concentration of individual glycoses in different strains was also noted. Mannose-KDO, galactose-KDO, and glucose-KDO ratios of virulent gonococci exceeded those of avirulent organisms, except that the correlation for glucose was not quite so striking. This relationship was not found in N. sicca and N. lactamica strains. Fatty acid analyses of lipid A from gonococci showed that 10-, 12-, 14-, 16-, and 18-carbon acids, as well as 3-hydroxytetradecanoic acid, were present, but differences in concentration between colonial types, although evident in some cases, appeared less significant than glycose content.

Wiseman GM; Caird JD

1977-05-01

236

Composition of the lipopolysaccharide of Neisseria gonorrhoeae.  

Science.gov (United States)

Analysis of glycose and fatty acid content of lipopolysaccharide extracted from 38 strains of Neisseria gonorrhoeae indicated that glycoses common to colonial types 1 to 5 were glucose, mannose, and galactose, N-acetylneuraminic acid, 2-keto-3-deoxyoctulosonic acid (KDO), glucosamine, and galactosamine were also invariably present. Virulent colonial types 1 and 2 contained no rhamnose, in contrast to avirulent types 3 to 5 and several strains of the nonpathogenic species N. sicca and N. lactamica. Fucose, characteristic of these nonpathogenic species, was not present in the gonococci. Variation in the concentration of individual glycoses in different strains was also noted. Mannose-KDO, galactose-KDO, and glucose-KDO ratios of virulent gonococci exceeded those of avirulent organisms, except that the correlation for glucose was not quite so striking. This relationship was not found in N. sicca and N. lactamica strains. Fatty acid analyses of lipid A from gonococci showed that 10-, 12-, 14-, 16-, and 18-carbon acids, as well as 3-hydroxytetradecanoic acid, were present, but differences in concentration between colonial types, although evident in some cases, appeared less significant than glycose content. PMID:405323

Wiseman, G M; Caird, J D

1977-05-01

237

Lipopolysaccharide Extracellular Emulsifier Produced by Penicillium citrinum  

Directory of Open Access Journals (Sweden)

Full Text Available A Brazilian strain of Penicillium citrinum produced a lipopolysaccharide with emulsifier properties during cultivation on mineral medium, containing ammonium sulfate as nitrogen source, with 1% (v/v) olive oil as the carbon source. The maximal emulsifier production (1.6 U mL-1) was obtained after 60 h of cultivation and the biomass reached 7.5 g L-1 at the end of fermentation. The production yield i.e. the amount the carbon source utilized for product synthesis was 54% and the best emulsifying activity was observed for xylene and diesel oil when compared to other carbohydrates tested. The emulsifier was shown to be stable to a wide range of pH and temperature values and was shown to contain D-galactose, D-glucose and D-xylose (8.2:1.0:5.3) with a total carbohydrate content of 43%. The presence of salts stimulated the emulsification activity, suggesting potential for its application in industrial waste or marine remediation.

M.M. Camargo de Morais; S.A.F. Ramos; M.C.B. Pimentel; E.H.M. Melo; M.A. Morais Jr; J.F. Kennedy; J.L. Lima Filho

2006-01-01

238

Cellular and free lipopolysaccharides of some species of Neisseria.  

UK PubMed Central (United Kingdom)

Cultures of eight non-pathogenic species of Neisseria grown in simple defined media released lipopolysaccharide (free lipopolysaccharide) by a process distinct from cellular autolysis. Analyses of the pure cellular and free lipopolysaccharides obtained from six species of Neisseria revealed that they were remarkably similar and were devoid of detectable O-antigen side chains. Three distinct types of core-oligosaccharides were demonstrated. Type I core-oligosaccharide was a branched structure of alpha-D-glucopyranosyl units (7 mol) terminated by a reducing end group of 3-deoxy-D-manno-octulosonic acid. Type II core-oligosaccharide contained D-glucose, 2-deoxy-2-amino-D-glucose, L-rhamnose, L-glycero-D-manno-heptose, 3-deoxy-D-manno-octulosonic acid, phosphate, and ethanolamine in a molar ratio of 3:2:1:1:1:1:1. Type III coreoligosaccharide was composed of D-glucose, L-glycero-D-manno-heptose, 3-deoxy-D-manno-octulosonic acid, and phosphate in a molar ratio of 3:3:1:1. Lipopolysaccharides of N. caviae and N. sicca contained type I core-oligosaccharides exclusively, while those of N. flava and N. perflava contained only type II core-oligosaccharide. Cellular lipopolysaccharide from N. cinerea contained core-oligosaccharides of types I and II in a ratio of 27:73, while the analogous preparation from N. flavescens contained core-oligosaccharide types II and III in a ratio of 21:4. Free lipopolysaccharides from these two organisms contained only one type of coreoligosaccharide. Lipid A components of all the lipopolysaccharide preparations were very similar being composed of about 25% by weight of dodecanoic acid, 3-hydroxy-dodecanoic acid, and 3-hydroxy-tetradecanoic acid.

Johnson KG; Perry MB; McDonald IJ; Russel RR

1975-12-01

239

Cellular and free lipopolysaccharides of some species of Neisseria.  

Science.gov (United States)

Cultures of eight non-pathogenic species of Neisseria grown in simple defined media released lipopolysaccharide (free lipopolysaccharide) by a process distinct from cellular autolysis. Analyses of the pure cellular and free lipopolysaccharides obtained from six species of Neisseria revealed that they were remarkably similar and were devoid of detectable O-antigen side chains. Three distinct types of core-oligosaccharides were demonstrated. Type I core-oligosaccharide was a branched structure of alpha-D-glucopyranosyl units (7 mol) terminated by a reducing end group of 3-deoxy-D-manno-octulosonic acid. Type II core-oligosaccharide contained D-glucose, 2-deoxy-2-amino-D-glucose, L-rhamnose, L-glycero-D-manno-heptose, 3-deoxy-D-manno-octulosonic acid, phosphate, and ethanolamine in a molar ratio of 3:2:1:1:1:1:1. Type III coreoligosaccharide was composed of D-glucose, L-glycero-D-manno-heptose, 3-deoxy-D-manno-octulosonic acid, and phosphate in a molar ratio of 3:3:1:1. Lipopolysaccharides of N. caviae and N. sicca contained type I core-oligosaccharides exclusively, while those of N. flava and N. perflava contained only type II core-oligosaccharide. Cellular lipopolysaccharide from N. cinerea contained core-oligosaccharides of types I and II in a ratio of 27:73, while the analogous preparation from N. flavescens contained core-oligosaccharide types II and III in a ratio of 21:4. Free lipopolysaccharides from these two organisms contained only one type of coreoligosaccharide. Lipid A components of all the lipopolysaccharide preparations were very similar being composed of about 25% by weight of dodecanoic acid, 3-hydroxy-dodecanoic acid, and 3-hydroxy-tetradecanoic acid. PMID:1220863

Johnson, K G; Perry, M B; McDonald, I J; Russel, R R

1975-12-01

240

Prenatal lipopolysaccharide increases maternal behavior, decreases maternal odor preference, and induces lipopolysaccharide hyporesponsiveness  

Scientific Electronic Library Online (English)

Full Text Available Abstract in english The present study investigated whether late maternal inflammation disrupts the mother/pup interaction, resulting in long-lasting effects on pup behavior and alterations in biological pathways, thereby programming prepubertal behavior and the pups' inflammatory responses after bacterial endotoxin treatment. Female rats received 100 ?g/kg lipopolysaccharide (LPS) or .9% saline solution on gestation day 18. Reproductive performance was observed at birth. On lactation da (more) ys (LD) 5 and LD 6, respectively, maternal behavior and maternal aggressive behavior were assessed. In pups, maternal odor preference on LD 7, open field behavior on LD 21, and serum tumor necrosis factor ? (TNF-?) levels after LPS challenge on LD 21 were investigated. The results showed that prenatal LPS exposure improved maternal care and reduced maternal aggressive behavior but did not alter maternal reproductive performance. Male offspring exhibited increased body weights at birth and reduced maternal odor preference. Lipopolysaccharide challenge increased the duration of immobility in the open field and induced a slight increase in serum TNF-? levels. Prenatal exposure to LPS during late pregnancy improved maternal care, reduced maternal olfactory preference, and induced TNF-? hyporesponsiveness to a single dose of LPS in pups.

Penteado, Sandra; Gomes, Cristina de Oliveira Massoco-Salles; Kirsten, Thiago; Reis-Silva, Thiago; Melo, Rafael César de; Acenjo, Michelli; Queiroz-Hazarbassanov, Nicolle; Bernardi, Maria Martha

2013-06-01

 
 
 
 
241

Eikenella corrodens and Porphyromonas asaccharolytica pleural empyema in a diabetic patient with obstructive sleep apnea syndrome on noninvasive ventilation.  

UK PubMed Central (United Kingdom)

Eikenella corrodens is a normal inhabitant of the human oral cavity and gastrointestinal and genitourinary tracts. Pleuropulmonary infections by this microorganism are uncommon. Pulmonary aspiration is a chief predisposing condition. Although the outcome is usually favorable, its distinctive antibiotic sensitivity pattern makes bacterial identification an important feature in dealing with this infection. The authors report a case of pleural empyema caused by co-infection with Eikenella corrodens and Porphyromonas asaccharolytica, in an immunocompetent diabetic patient with obstructive sleep apnea syndrome, followed by a discussion on the role of noninvasive ventilation in the development of this infection.

Caiano Gil J; Calisto R; Amado J; Barreto V

2013-03-01

242

Molecular mechanism in tolerance to lipopolysaccharide.  

UK PubMed Central (United Kingdom)

Stimulation with lipopolysaccharide (LPS) will lead to the expression of a variety of genes in CD14+ monocytes/macrophages, but also in CD14- fibroblasts and endothelial cells. Upon secondary LPS stimulation, the expression of many of these genes is only minimal. This applies to several cytokines, most prominent among them tumor necrosis factor (TNF). Induction of tolerance appears to require some degree of activation in the primary exposure, as partial structures of LPS induce tolerance, as long as they are able to activate cells. Studies on the mechanism of unresponsiveness in tolerant cells show that the CD14 LPS receptor is not downregulated but may even increase in number at the cell surface. Furthermore, this receptor appears to be functional in that mobilization of the transcription factor NF-kappa B does still occur. This NF-kappa B complex is composed primarily of p50p50 homodimers, that bind to the respective DNA motif in the promoter region of many proinflammatory genes, thereby blocking transactivation. However, LPS tolerance does not lead to downregulation of all kinds of response, as some genes are even increased in expression upon secondary stimulation; these include p50 of NF-kappa B, TNF receptor type II and interleukin-10 (IL-10). These gene products are involved in the downregulation of proinflammatory cytokines and may thereby be instrumental in the unresponsiveness observed. Hence, tolerance to LPS is not a passive process that occurs in an exhausted cell; rather, it is a well-controlled active response that is orchestrated in order to prevent excessive inflammation. Important modulators of tolerance are glucocorticoids, which result in a general decrease of gene expression, and interferon-gamma (IFN-gamma), which enhances expression of proinflammatory genes. LPS tolerance does occur in some clinical settings, as in hemodialysis, in sepsis and in patients treated repeatedly with LPS or other monocyte activators. In fact, LPS tolerance may be exploited for prophylaxis of severe sepsis in patients at risk.

Ziegler-Heitbrock HW

1995-01-01

243

Curcumin attenuates lipopolysaccharide-induced renal inflammation.  

UK PubMed Central (United Kingdom)

Renal inflammation is the main pathological change in many acute and chronic kidney diseases. Curcumin, a yellow pigment present in the rhizome of turmeric (Curcuma longa L. Zingiberaceae), was found to be a potential anti-inflammatory agent. The present study aimed to investigate the effects of curcumin on the inflammation of mice kidney and cultured renal tubular epithelial cells (HK-2 cells) induced by lipopolysaccharide (LPS) and to explore the mechanism. Curcumin was injected intraperitoneally before LPS administration. Renal inflammation was assessed by evaluating monocyte chemoattractant protein-1 (MCP-1) expression and macrophage infiltration in renal tissue using immunohistochemical methods, and also by measuring renal MCP-1 mRNA level using Real-Time polymerase chain reaction (PCR). HK-2 cells were cultured to investigate the in vitro effect of curcumin against LPS-induced renal inflammation. The expression of MCP-1 and interleukin-8 (IL-8) mRNA was measured by Real-Time PCR. The expression of MCP-1 and IL-8 protein in supernatant was detected by enzyme-linked immunosorbent assay (ELISA). The activity of nuclear factor (NF)-?B was detected by electrophoretic mobility shift assay (EMSA). The results demonstrated that curcumin could inhibit LPS-induced renal MCP-1 mRNA expression. Curcumin also significantly inhibited the expression of MCP-1 and IL-2 mRNA in HK-2 cells, and partially inhibited the secretion of MCP-1 and IL-8. Furthermore, curcumin was found to inhibit the DNA-binding activity of NF-?B. The present study demonstrated that curcumin has a protective effect on LPS-induced experimental renal inflammation, and this effect might be attributed to its inhibitory effects on MCP-1 mRNA expression and DNA-binding activity of NF-?B. Hence, curcumin might be potentially useful in some kidney diseases by preventing renal inflammation.

Zhong F; Chen H; Han L; Jin Y; Wang W

2011-01-01

244

Lipopolysaccharide tolerance inhibits eye inflammation. I. Reduced immune complex or lipopolysaccharide effects.  

UK PubMed Central (United Kingdom)

The effect of endotoxin tolerance on ocular inflammation was studied in rabbits. A single intravenous (IV) injection of endotoxin (bacterial lipopolysaccharide [LPS]) produced a mild acute iridocyclitis. Repeated daily (five to seven days) IV injections of LPS (5 micrograms extracted from Salmonella typhimurium) led to a state of refractoriness or LPS "tolerance," and ocular inflammation was no longer produced. In contrast to controls, in rabbits tolerant to LPS, IV LPS failed to elevate prostaglandin E2, thromboxane B2, or chemotactic factors in the aqueous humor. Rabbits tolerant to LPS also resisted the increase in vascular permeability normally induced by an ocular reversed passive Arthus reaction. These results demonstrated that LPS tolerance can induce anti-inflammatory effects in the eye.

Howes EL Jr; Goldyne ME; Perez HD; Goldstein IM; Rosenbaum JT

1985-02-01

245

The effect of 2% chlorhexidine digluconate irrigation on clinical parameters and the level of Bacteroides gingivalis in periodontal pockets.  

UK PubMed Central (United Kingdom)

Eight patients with moderate periodontitis volunteered to participate in a study to assess the effect of subgingival 2% chlorhexidine irrigation, with and without scaling and root planing, on clinical parameters and the level of Bacteroides gingivalis in periodontal pockets. Each quadrant was required to have at least one site with a probing depth of 6 mm or greater and bleeding on probing. The patients were treated following a randomized four quadrant design: one quadrant received no treatment; a second quadrant received scaling and root planing only; a third quadrant received chlorhexidine irrigation only; the fourth quadrant received scaling and root planing, plus chlorhexidine irrigation. Sites to receive chlorhexidine were irrigated at 0, 1, 2, and 3 weeks. Clinical and microbiological indices were measured and recorded at 0, 5, 7, 11, and 15 weeks. The clinical parameters measured included; Plaque Index (PI), Gingival Index (GI), probing depth (PD), Bleeding Tendency (BT), and attachment level (AL). The attachment level was measured using an occlusal stint as a fixed reference point. The level of Bacteroides gingivalis was measured by labeling the plaque sample with a polyclonal fluorescent antibody. The plaque smear was then read using a fluorescent microscope at 1000 magnification. The Spearman Rank-Order Correlation was used to determine the relationship between parameters at baseline. The effects of the treatment groups were compared using the Neuman-Keuls Multiple Comparison Technique. The results showed that a positive correlation existed between B. gingivalis (rs = 0.68) and Bleeding Tendency and between P1I (rs = 0.77) and GI.(ABSTRACT TRUNCATED AT 250 WORDS)

Southard SR; Drisko CL; Killoy WJ; Cobb CM; Tira DE

1989-06-01

246

Efectos de un gel de tetraciclina al 5% sobre los niveles de P. gingivalis, P. intermedia y A. actinomycetemcomitans  

Scientific Electronic Library Online (English)

Full Text Available Abstract in spanish El objetivo de la presente investigación fue estudiar los efectos de un gel de tetraciclina al 5% sobre los niveles de 3 microorganismos asociados al desarrollo de la periodontitis rápidamente progresiva (PRP). En un total de 20 pacientes con PRP se seleccionaron 5 dientes por paciente con bolsas periodontales > a 5 mm. con sangrado al sondaje periodontal en los cuales se efectuaron los siguientes tratamientos: 1. Control. 2. Aplicación de un gel de tetraciclina al 5% (more) (T). 3. Placebo (Pl). 4. Raspado y alisado radicular (RA). 5. T + RA .De forma previa, en cada sitio seleccionado, se tornaron muestras de placa subgingival con un cono de papel estéril para detectar y cuantificar la presencia de P. gingivalis , P. intermedia y A. actinomycetemcomitans, mediante el uso de sondas DNA (OMNIGENE, U.S.A.).Se dieron instrucciones de higiene oral, y se efectuó un nuevo control microbiológico a los 60 días El análisis estadístico de los resultados demostró lo siguiente: 1. Ninguno de los tratamientos redujo significativamente los niveles de A. actinomycetemcomitans. 2. Se detectó una reducción significativa de P. gingivalis en los sitios tratados con (R.A). (p Abstract in english The purpose of this investigation was to study the effects of local delivery of a tetracycline 5% gel on the levels of 3 bacteria associated to development of rapidly progressive periodontitis. In a sample of 20 patients, five teeth were selected from each patient with periodontal pockets > 5 mm and bleeding upon probing. One of the following treatments were done at each selected site: 1. Control (no treatment). 2. Local delivery of a 5% tetracycline gel. 3. Placebo gel. (more) 4. Scaling and root planing. 5. Scaling and root planing + local application of tetracycline gel. Previously, at each selected site samples of subgingival plaque were taken with sterile paper points in order to detecte and quantify the presence of P.gingivalis, P.intermedia and A. actinomycetemcomitans, by using DNA probe technology (OMNIGENE, U.S.A.). Oral hygiene instructions were given to each patient and new samples of subgingival plaque were obtained at 60 days. Statistical analysis of results showed the following 1. No significant reductions of A. actinomycetemcomitans were found with performed treatments. 2. Scaling and root planing reduced the levels of P. gingivalis (p 0.02) or when the later was combined with tetracycline (p

Gallardo, F.; Plaza, J.C.; Sotta, R. de la

2002-04-01

247

Characterization of the lipopolysaccharide from Vibrio cholerae 395 (Ogawa).  

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The chemical structure and biological properties of the lipopolysaccharide (LPS) from Vibrio cholerae 395 (Ogawa), isolated by the phenol-water procedure, were studied. Upon acid hydrolysis, the LPS was split into its polysaccharide and lipid A moieties. The polysaccharide contained both neutral (gl...

Kabir, S

248

Induction of oral tolerance in mice unresponsive to bacterial lipopolysaccharide.  

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C3H/HeJ lipopolysaccharide (LPS)-unresponsive and closely related C3H/HeSn LPS-responsive mice were rendered tolerant to hen albumin by antigen feeding before parenteral immunization. Both single and multiple feedings of antigen were effective in inducing tolerance to hen albumin in LPS-responsive a...

Saklayen, M G; Pesce, A J; Pollak, V E; Michael, J G

249

Fosfomycin Alters Lipopolysaccharide-Induced Inflammatory Cytokine Production in Mice  

Science.gov (United States)

To determine the mechanisms of immunomodulating action of fosfomycin (FOF), we examined its effect on the production of inflammatory cytokines in mice injected with lipopolysaccharide (LPS). Treatment with FOF significantly lowered the peak serum levels of tumor necrosis factor alpha and interleukin-1?, indicating that FOF alters inflammatory cytokine production after LPS stimulation.

Matsumoto, Tetsuya; Tateda, Kazuhiro; Miyazaki, Shuichi; Furuya, Nobuhiko; Ohno, Akira; Ishii, Yoshikazu; Hirakata, Yoichi; Yamaguchi, Keizo

1999-01-01

250

Differential regulation of cytokine production in lipopolysaccharide tolerance in mice.  

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We investigated the pattern of down-regulation of cytokine production in endotoxin (lipopolysaccharide [LPS]) tolerance. A 4-day treatment with LPS (35 micrograms per mouse) was followed by a challenge on day 6 with one more injection of LPS. Circulating tumor necrosis factor (TNF) and interleukin-6...

Erroi, A; Fantuzzi, G; Mengozzi, M; Sironi, M; Orencole, S F; Clark, B D; Dinarello, C A; Isetta, A; Gnocchi, P; Giovarelli, M

251

Acetaminophen reduces lipopolysaccharide-induced fever by inhibiting cyclooxygenase-2.  

UK PubMed Central (United Kingdom)

Acetaminophen is one of the world's most commonly used drugs to treat fever and pain, yet its mechanism of action has remained unclear. Here we tested the hypothesis that acetaminophen blocks fever through inhibition of cyclooxygenase-2 (Cox-2), by monitoring lipopolysaccharide induced fever in mice with genetic manipulations of enzymes in the prostaglandin cascade. We exploited the fact that lowered levels of a specific enzyme make the system more sensitive to any further inhibition of the same enzyme. Mice were immune challenged by an intraperitoneal injection of bacterial wall lipopolysaccharide and their body temperature recorded by telemetry. We found that mice heterozygous for Cox-2, but not for microsomal prostaglandin E synthase-1 (mPGES-1), displayed attenuated fever, indicating a rate limiting role of Cox-2. We then titrated a dose of acetaminophen that did not inhibit the lipopolysaccharide-induced fever in wild-type mice. However, when the same dose of acetaminophen was given to Cox-2 heterozygous mice, the febrile response to lipopolysaccharide was strongly attenuated, resulting in an almost normalized temperature curve, whereas no difference was seen between wild-type and heterozygous mPGES-1 mice. Furthermore, the fever to intracerebrally injected prostaglandin E2 was unaffected by acetaminophen treatment. These findings reveal that acetaminophen, similar to aspirin and other non-steroidal anti-inflammatory drugs, is antipyretic by inhibiting cyclooxygenase-2, and not by inhibiting mPGES-1 or signaling cascades downstream of prostaglandin E2.

Engström Ruud L; Wilhelms DB; Eskilsson A; Vasilache AM; Elander L; Engblom D; Blomqvist A

2013-08-01

252

Sphingosine Kinase Protects Lipopolysaccharide-Activated Macrophages from Apoptosis  

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Lipopolysaccharide (LPS) signaling is critical for the innate immune response to gram-negative bacteria. Here, evidence is presented for LPS stimulation of sphingosine kinase (SPK) in the RAW 264.7 murine macrophage cell line and rat primary hepatic macrophages (HMs). LPS treatment of RAW 264.7 cell...

Wu, Weicheng; Mosteller, Raymond D.; Broek, Daniel

253

Acetaminophen reduces lipopolysaccharide-induced fever by inhibiting cyclooxygenase-2.  

Science.gov (United States)

Acetaminophen is one of the world's most commonly used drugs to treat fever and pain, yet its mechanism of action has remained unclear. Here we tested the hypothesis that acetaminophen blocks fever through inhibition of cyclooxygenase-2 (Cox-2), by monitoring lipopolysaccharide induced fever in mice with genetic manipulations of enzymes in the prostaglandin cascade. We exploited the fact that lowered levels of a specific enzyme make the system more sensitive to any further inhibition of the same enzyme. Mice were immune challenged by an intraperitoneal injection of bacterial wall lipopolysaccharide and their body temperature recorded by telemetry. We found that mice heterozygous for Cox-2, but not for microsomal prostaglandin E synthase-1 (mPGES-1), displayed attenuated fever, indicating a rate limiting role of Cox-2. We then titrated a dose of acetaminophen that did not inhibit the lipopolysaccharide-induced fever in wild-type mice. However, when the same dose of acetaminophen was given to Cox-2 heterozygous mice, the febrile response to lipopolysaccharide was strongly attenuated, resulting in an almost normalized temperature curve, whereas no difference was seen between wild-type and heterozygous mPGES-1 mice. Furthermore, the fever to intracerebrally injected prostaglandin E2 was unaffected by acetaminophen treatment. These findings reveal that acetaminophen, similar to aspirin and other non-steroidal anti-inflammatory drugs, is antipyretic by inhibiting cyclooxygenase-2, and not by inhibiting mPGES-1 or signaling cascades downstream of prostaglandin E2. PMID:23545161

Engström Ruud, Linda; Wilhelms, Daniel Björk; Eskilsson, Anna; Vasilache, Ana Maria; Elander, Louise; Engblom, David; Blomqvist, Anders

2013-03-29

254

Role of calcium during lipopolysaccharide stimulation of neutrophils.  

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This study investigated the role of intracellular calcium concentration ([Ca]i) as a possible intermediate in the lipopolysaccharide (LPS) second messenger pathway for the activation of neutrophils (polymorphonuclear leukocytes [PMNs]). Isolated PMNs were loaded with the calcium-sensitive fluorescen...

Rodeberg, D A; Babcock, G F

255

THE IMPORTANCE OF AGGREGATIBACTER ACTINOMYCETEMCOMITANS IN ETIOLOGY OF PERIODONTAL DISEASE - MINI REVIEW  

Directory of Open Access Journals (Sweden)

Full Text Available Periodontal disease is a chronic, degenerative disease of parodontium which is made of gingiva, periodontal ligament, cementum and alveolar bone. The main etiological factor for development of periodontal disease is dental plaque or oral biofilm in association with anaerobic bacteria. Aggregatibacter actinomycetemcomitans is one of the most powerful periodonthopathogens. This microorganism produces many virulent factors: leucotoxin as the most important, then bacteriocin, chemotaxis inhibiting factor, cytotoxic factors, Fc binding proteins, immunosuppressive factors, lipopolysaccharide collagenase, fibroblast inhibiting factor, antibiotic resistance determinants, adhesives, invasives and function inhibiting factor of polymorphonuclear leukocytes. The ability of Aggregatibacter actinomycetemcomitans lipopolysaccharides to stimulate macrophages to release interleukins IL-1, IL-1?, and tumor necrosis factor (TNF) is of main importance. These cytokines are able to stimulate the bone resorption. Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis represent exogenous microorganisms, based on its minor presence in healthy individuals. It has been recommended that periodontal diseases associated with periodontal pathogens represent "true infections".

Ljiljana Kesi?; Milica Petrovi?; Radmila Obradovi?; Ana Pej?i?

2009-01-01

256

Discrimination between Bacteroides, Dichelobacter, Fusobacterium, Porphyromonas and Prevotella isolated from caprine footrot by PCR-RFLP - short communication.  

Science.gov (United States)

Footrot is widely considered the most severe and most common foot pathology in small ruminants. This study tested the ability of a molecular typing system based on polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay of the 16S rRNA gene to discriminate between the strict anaerobe genera most commonly isolated from footrot ( Bacteroides, Dichelobacter, Fusobacterium, Porphyromonas and Prevotella ) in goats in Extremadura (Spain), with a view to facilitating identification for diagnostic purposes and thus providing a useful tool for future epidemiological studies. Although the electrophoretic patterns obtained with the enzyme Tru 1I were more readily interpreted, and may thus be the best initial option, results may be confirmed by a second enzyme ( RsaI). The PCR-RFLP assay of the 16S rRNA gene may therefore prove a useful addition to conventional biochemical identification techniques, providing taxonomic information at genus level. PMID:19584033

Lacombe-Antoneli, Angela; Píriz, Segundo; Quesada, Alberto; Vadillo, Santiago

2009-06-01

257

The alteration of copper homeostasis in inflammation induced by lipopolysaccharides.  

UK PubMed Central (United Kingdom)

Significant changes of copper homeostasis were triggered by lipopolysaccharides, which result in systemic inflammatory response and contribute to hepatic injury. Administration of lipopolysaccharides resulted in the increase of plasma "free" copper and total copper concentrations, whereas, the decrease of "free" copper and total copper contents in liver tissue. Copper-associated proteins were detected and showed a down-regulation of X-linked inhibitor of apoptosis protein, and up-regulation of copper metabolism domain containing 1 and copper transporter 1. The alteration of these proteins would lower the apoptotic threshold. Meanwhile, the increasing of circulation copper might cause oxidative injury through Fenton reaction and contribute to tissue injury. Our findings underscored the possibility that these changes in systemic copper homeostasis might provide a novel insight of the characteristic of the acute phase of inflammatory response and the underlying influence on tissue injury.

Han M; Lin Z; Zhang Y

2013-08-01

258

Innate immunity probed by lipopolysaccharides affinity strategy and proteomics.  

UK PubMed Central (United Kingdom)

Lipopolysaccharides (LPSs) are ubiquitous and vital components of the cell surface of Gram-negative bacteria that have been shown to play a relevant role in the induction of the immune-system response. In animal and plant cells, innate immune defenses toward microorganisms are triggered by the perception of pathogen associated molecular patterns. These are conserved and generally indispensable microbial structures such as LPSs that are fundamental in the Gram-negative immunity recognition. This paper reports the development of an integrated strategy based on lipopolysaccharide affinity methodology that represents a new starting point to elucidate the molecular mechanisms elicited by bacterial LPS and involved in the different steps of innate immunity response. Biotin-tagged LPS was immobilized on streptavidin column and used as a bait in an affinity capture procedure to identify protein partners from human serum specifically interacting with this effector. The complex proteins/lipopolysaccharide was isolated and the protein partners were fractionated by gel electrophoresis and identified by mass spectrometry. This procedure proved to be very effective in specifically binding proteins functionally correlated with the biological role of LPS. Proteins specifically bound to LPS essentially gathered within two functional groups, regulation of the complement system (factor H, C4b, C4BP, and alpha 2 macroglobulin) and inhibition of LPS-induced inflammation (HRG and Apolipoproteins). The reported strategy might have important applications in the elucidation of biological mechanisms involved in the LPSs-mediated molecular recognition and anti-infection responses.

Giangrande C; Colarusso L; Lanzetta R; Molinaro A; Pucci P; Amoresano A

2013-01-01

259

Endotoxin tolerance of adrenal gland: attenuation of corticosterone production in response to lipopolysaccharide and adrenocorticotropic hormone.  

UK PubMed Central (United Kingdom)

OBJECTIVES: Reversible adrenal insufficiency frequently has been diagnosed in critically ill patients with sepsis who have either low basal cortisol levels or low cortisol responses to adrenocorticotropic hormone (ACTH) stimulation. It is generally accepted that a phenomenon called "endotoxin tolerance" contributes to immunosuppression during sepsis. The present study was to investigate whether endotoxin tolerance occurs in the adrenal gland, leading to hyporesponsiveness of adrenal gland during sepsis. DESIGN: Controlled laboratory experiment. SETTING: University research laboratory. SUBJECTS: Sprague-Dawley male rats 200-250 g and primary isolated adrenal fasciculata-reticularis cells. INTERVENTIONS: Rats received intra-arterial injection of purified lipopolysaccharide (0.5 mg/kg) through indwelling femoral arterial catheters, and 24 hrs later the adrenocortical sensitivity to exogenous ACTH (10 ng/kg) was detected. Primary fasciculata-reticularis cells were pretreated with lipopolysaccharide at 0.1-100 ng/mL or with ACTH at 0.01-10 ng/mL and then challenged, in fresh media, with 1 ?g/mL lipopolysaccharide or 10 ng/mL ACTH. MEASUREMENTS AND MAIN RESULTS: Toll-like receptor 4 was expressed in adrenal gland and primary fasciculata-reticularis cells. Plasma corticosterone response to ACTH was decreased in rats receiving preinjection of lipopolysaccharide. Lipopolysaccharide pretreatment caused a significant decrease in corticosterone production in response to subsequent ACTH and lipopolysaccharide stimulation in primary fasciculata-reticularis cells. Lipopolysaccharide pretreatment inhibited ACTH- and lipopolysaccharide-induced expression of steroid metabolizing enzymes. Lipopolysaccharide significantly decreased Toll-like receptor 4 and ACTH receptor expression. CONCLUSIONS: Pre-exposure to lipopolysaccharide resulted in hyporesponsiveness to ACTH stimulation in rats. In vitro, lipopolysaccharide pretreatment impaired corticosterone production of fasciculata-reticularis cells in response to ACTH and lipopolysaccharide, which was associated with decreased expression of synthetic enzymes required for corticosterone production. Our results indicate that endotoxin tolerance of adrenal gland is one of the mechanisms for adrenocortical insufficiency during sepsis.

Liu S; Zhu X; Liu Y; Wang C; Wang S; Tang X; Ni X

2011-03-01

260

Meningo-encefalite equina da Halicephalobus gingivalis: contributo casistico nell’ambito delle attività di sorveglianza della Febbre del Nilo occidentale (West Nile disease)  

Directory of Open Access Journals (Sweden)

Full Text Available Un cavallo di 7 anni è stato abbattuto dopo aver manifestato una grave sindrome neurologica a rapida evoluzione. Campioni tessutali sono stati inviati al Centro Studi Malattie Esotiche dell’Istituto Zooprofilattico Sperimentale dell’Abruzzo e del Molise “G. Caporale” (Istituto G. Caporale) per gli accertamenti diagnostici del caso. Gli esami per le più comuni virosi neurologiche equine non hanno evidenziato la presenza di infezioni in atto. Istologicamente, si è osservata a livello encefalico la presenza di manicotti perivascolari e numerosi corpi parassitari, morfologicamente riferibili a Halicephalobus gingivalis. Il rinvenimento ha consentito di formulare la diagnosi di meningo-encefalite da H. gingivalis. Il caso riportato conferma che le encefaliti parassitarie devono essere annoverate nella diagnosi differenziale delle encefalopatie equine e sottolinea l’utilità dell’approccio diagnostico multidisciplinare.

Gabriella Di Francesco; Giovanni Savini; Alessandro Maggi; Nicola Cavaliere; Anna Rita D’Angelo; Giuseppe Marruchella

2012-01-01

 
 
 
 
261

Computer Simulation of the Rough Lipopolysaccharide Membrane of Pseudomonas Aeruginosa  

Energy Technology Data Exchange (ETDEWEB)

Lipopolysaccharides (LPS) form the major constituent of the outer membrane of Gram-negative bacteria, and are believed to play a key role in processes that govern microbial metal binding, microbial adsorption to mineral surfaces, and microbe mediated oxidation/reduction reactions at the bacterial exterior surface. A computational modeling capability is being developed for the study of geochemical reactions at the outer bacterial envelope of Gram-negative bacteria. The understanding of these mechanisms is crucial for the development of successful environmental bioremediation strategies. A molecular model for the rough LPS of Pseudomonas aeruginosa has been designed based on available experimentally determined structural information.

Lins, Roberto D.; Straatsma, TP

2001-08-01

262

Inhibition of lipopolysaccharide (LPS)-induced endothelial cytotoxicity by diosmin.  

UK PubMed Central (United Kingdom)

The cytotoxic effect of lipopolysaccharide (LPS) on cultivated bovine aortic endothelial cells was determined. This LPS-induced cytotoxicity was attenuated by diosmin. That means, the IC50 value of LPS in the combination with 8 mumol/l diosmin was shifted from 31 to 70 ng/ml in a concentration dependent manner. As a hypothesis it was suggested, that the inhibition of LPS-induced cytotoxicity in bovine aortic endothelial cell cultures by diosmin could be probably mediated via inhibition of tyrosine kinases.

Melzig MF; Loose R

1999-04-01

263

Structure of the core oligosaccharide in the serotype O8 lipopolysaccharide from Klebsiella pneumoniae.  

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Two classes of mutants with O-antigen-deficient lipopolysaccharides were isolated from the serotype O8 reference strain, belonging to Klebsiella pneumoniae subspecies ozaenae. These mutants were selected by resistance to bacteriophage KO1-2, which recognizes and lyses strains with lipopolysaccharide...

Severn, W B; Kelly, R F; Richards, J C; Whitfield, C

264

Detection of lipopolysaccharides by ethidium bromide staining after sodium dodecyl sulfate-polyacrylamide gel electrophoresis.  

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A rapid and easy method for staining lipopolysaccharides with ethidium bromide is described. Lipopolysaccharides could be visualized by ethidium bromide with almost the same sensitivity as found with the silver-staining method in less than 30 min. The ethidium bromide-staining method was particularl...

Kido, N; Ohta, M; Kato, N

265

Lysine-spermine conjugates: hydrophobic polyamine amides as potent lipopolysaccharide sequestrants.  

UK PubMed Central (United Kingdom)

Lipopolysaccharides (LPS), otherwise termed 'endotoxins', are outer-membrane constituents of Gram-negative bacteria. Lipopolysaccharides play a key role in the pathogenesis of 'Septic Shock', a major cause of mortality in the critically ill patient. Therapeutic options aimed at limiting downstream systemic inflammatory processes by targeting lipopolysaccharide do not exist at the present time. We have defined the pharmacophore necessary for small molecules to specifically bind and neutralize LPS and, using animal models of sepsis, have shown that the sequestration of circulatory LPS by small molecules is a therapeutically viable strategy. In this paper, the interactions of a focused library of lysine-spermine conjugates with lipopolysaccharide (LPS) have been characterized. Lysine-spermine conjugates with the epsilon-amino terminus of the lysinyl moiety derivatized with long-chain aliphatic hydrophobic substituents in acyl or alkyl linkage bind and neutralize bacterial lipopolysaccharides, and may be of use in the prevention or treatment of endotoxic shock states.

Burns MR; Wood SJ; Miller KA; Nguyen T; Cromer JR; David SA

2005-04-01

266

Prolonged sleep fragmentation of mice exacerbates febrile responses to lipopolysaccharide.  

UK PubMed Central (United Kingdom)

BACKGROUND: Sleep disruption is a frequent occurrence in modern society. Whereas many studies have focused on the consequences of total sleep deprivation, few have investigated the condition of sleep disruption. NEW METHOD: We disrupted sleep of mice during the light period for 9 consecutive days using an intermittently rotating disc. RESULTS: Electroencephalogram (EEG) data demonstrated that non-rapid eye movement (NREM) sleep was severely fragmented and REM sleep was essentially abolished during the 12h light period. During the dark period, when sleep was not disrupted, neither NREM sleep nor REM sleep times differed from control values. Analysis of the EEG revealed a trend for increased power in the peak frequency of the NREM EEG spectra during the dark period. The fragmentation protocol was not overly stressful as body weights and water consumption remained unchanged, and plasma corticosterone did not differ between mice subjected to 3 or 9 days of sleep disruption and home cage controls. However, mice subjected to 9 days of sleep disruption by this method responded to lipopolysaccharide with an exacerbated febrile response. COMPARISON WITH EXISTING METHODS: Existing methods to disrupt sleep of laboratory rodents often subject the animal to excessive locomotion, vibration, or sudden movements. This method does not suffer from any of these confounds. CONCLUSIONS: This study demonstrates that prolonged sleep disruption of mice exacerbates febrile responses to lipopolysaccharide. This device provides a method to determine mechanisms by which chronic insufficient sleep contributes to the etiology of many pathologies, particularly those with an inflammatory component.

Ringgold KM; Barf RP; George A; Sutton BC; Opp MR

2013-09-01

267

Neutralization and transfer of lipopolysaccharide by phospholipid transfer protein.  

UK PubMed Central (United Kingdom)

Phospholipid transfer protein (PLTP) and lipopolysaccharide-binding protein (LPB) are lipid transfer proteins found in human plasma. PLTP shares 24% sequence similarity with LBP. PLTP mediates the transfer and exchange of phospholipids between lipoprotein particles, whereas LBP transfers bacterial lipopolysaccharide (LPS) either to lipoprotein particles or to CD14, a soluble and cell-surface receptor for LPS. We asked whether PLTP could interact with LPS and mediate the transfer of LPS to lipoproteins or to CD14. PLTP was able to bind and neutralize LPS: incubation of LPS with purified recombinant PLTP (rPLTP) resulted in the inhibition of the ability of LPS to stimulate adhesive responses of neutrophils, and addition of rPLTP to blood inhibited cytokine production in response to LPS. Transfer of LPS by rPLTP was examined using fluorescence dequenching experiments and native gel electrophoresis. The results suggested that rPLTP was able to mediate the exchange of LPS between micelles and the transfer of LPS to reconstituted HDL particles, but it did not transfer LPS to CD14. Consonant with these findings, rPLTP did not mediate CD14-dependent adhesive responses of neutrophils to LPS. These results suggest that while PLTP and LBP both bind and transfer LPS, PLTP is unable to transfer LPS to CD14 and thus does not mediate responses of cells to LPS.

Hailman E; Albers JJ; Wolfbauer G; Tu AY; Wright SD

1996-05-01

268

Dexmedetomidine attenuates lipopolysaccharide-induced proinflammatory response in primary microglia.  

UK PubMed Central (United Kingdom)

BACKGROUND: Neuroinflammation mediated by microglia has been implicated in delirium. Suppression of microglial activation may therefore contribute to alleviate delirium. It has been reported that dexmedetomidine (DEX) has a potent anti-inflammatory property. In the present study, we investigated the effects of DEX on the production of proinflammatory mediators in lipopolysaccharide-stimulated microglia. MATERIALS AND METHODS: The concentrations of DEX were chosen to correspond to 1, 10, and 100 times of clinically relevant concentration (i.e., 1, 10, and 100ng/mL). The levels of proinflammatory mediators, such as inducible nitric oxide synthase or nitric oxide, prostaglandin E(2), interleukin 1?, and tumor necrosis factor ?, were measured. RESULTS: DEX at 1ng/mL did not affect the production of proinflammatory mediators. DEX at 10 and 100ng/mL significantly inhibited the release of nitric oxide, prostaglandin E(2), interleukin 1?, and tumor necrosis factor ? and the expression of inducible nitric oxide synthase messenger RNA. CONCLUSIONS: These results suggest that DEX is a potent suppressor of lipopolysaccharide-induced inflammation in activated microglia and may be a potential therapeutic agent for the treatment of intensive care unit delirium.

Peng M; Wang YL; Wang CY; Chen C

2013-01-01

269

Structure of the core oligosaccharide in the serotype O8 lipopolysaccharide from Klebsiella pneumoniae.  

Science.gov (United States)

Two classes of mutants with O-antigen-deficient lipopolysaccharides were isolated from the serotype O8 reference strain, belonging to Klebsiella pneumoniae subspecies ozaenae. These mutants were selected by resistance to bacteriophage KO1-2, which recognizes and lyses strains with lipopolysaccharide molecules containing the D-galactan II O antigen. Strain RFK-11 contains a defect in O-antigen synthesis and has a complete core, including the attachment site for O antigen. This mutation is complemented by a plasmid carrying the rfb (O-antigen biosynthesis) gene cluster from the related K. pneumoniae serotype O1. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the lipopolysaccharide from strain RFK-9 has a mobility typical of deep-rough lipopolysaccharide. RFK-9 lipopolysaccharide lacks the attachment site for O antigen. Lipopolysaccharides from strains RFK-9 and RFK-11 were isolated, and their structures were determined by methylation analyses, muclear magnetic resonance spectroscopy, and mass spectroscopy. The deduced O8 core oligosaccharide includes the partial core structure reported for the K. pneumoniae subspecies pneumoniae serotype O1 lipopolysaccharide (M. Süsskind, S. Müller-Leonnies, W. Nimmich, H. Brade, and O. Holst, Carbohydr. Res. 269:C1-7, 1995), consistent with the possibility of a conserved core structure within the species. The core oligosaccharide differs from those of the genera Salmonella and Escherichia by the absence of a hexose-containing outer core, the lack of phosphate residues in the inner core, and the presence of galacturonic acid residues. PMID:8626303

Severn, W B; Kelly, R F; Richards, J C; Whitfield, C

1996-03-01

270

DNA probe detection of periodontopathogens in advanced periodontitis.  

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Species-specific DNA probes were used to determine the presence of Actinobacillus actinomycetemcomitans (A.a.), Porphyromonas (Bacteroides) gingivalis, Prevotella intermedia, Treponema denticola, Eikenella corrodens, Fusobacterium nucleatum, and Wolinella recta in subgingival plaque from deep pocket...

Söder, PO; Jin, LJ; Söder, B

271

Antimicrobial activity of traditional Chinese medicines on common oral bacteria  

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Objective: To evaluate twenty Traditional Chinese Medicines (TCM) against four oral bacteria. Methods: Twenty TCM were tested for sensitivity against Streptococcus mitis, Streptococcus sanguis, Streptococcus mutans and Porphyromonas gingivalis. Aliquots of suspension of each bacterial species were i...

Yuen, MKZ; Wong, RWK; Hagg, U; Samaranayake, L

272

Antimicrobial Activity of Traditional Chinese Medicines on Common Oral Bacteria  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Objective: To evaluate twenty Traditional Chinese Medicines (TCM) against four oral bacteria. Methods: Twenty TCM were tested for sensitivity against Streptococcus mitis, Streptococcus sanguis, Streptococcus mutans and Porphyromonas gingivalis. Aliquots of suspension of each bacterial species were i...

Michelle K. Z. Yuen; Ricky W. K. Wong; Urban Hägg; Lakshman Samaranayake

273

Basic peptide protamine exerts antimicrobial activity against periodontopathic bacteria——Growth inhibition of periodontopathic bacteria by protamine  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Protamine was investigated for its antibacterial activity against the periodontal pathogens, Porphyromonas gingivalis, Prevotella intermedia and Aggregatibacter actinomycetemcomitans. We determined the minimum inhibitory concentrations of protamine and its hydrolysate and their bactericidal activity...

Tadashi Miura; Keishi Iohara; Tetsuo Kato; Kazuyuki Ishihara; Masao Yoshinari

274

Lipopolysaccharide enhances bactericidal activity in Dictyostelium discoideum cells.  

Science.gov (United States)

Innate immune cells respond to invading microbes upon detection of pathogen-associated molecular patterns (PAMPS). PAMP-recognition machinery is evolutionarily conserved, allowing for characterization in model organisms. The model organism Dictyostelium discoideum can exist as single-celled amoebae, which phagocytize bacteria for nutrients. Although D. discoideum is used extensively to study phagocytosis, it has not been determined if D. discoideum detects bacterial PAMPs using pattern-recognition machinery. Here we show that D. discoideum mounts responses against the bacterial cell wall PAMP, lipopolysaccharide (LPS). Upon treatment with LPS or its active component Lipid A, D. discoideum cells more efficiently clear phagocytized bacteria. LPS-enhanced bactericidal activity appears dependent both on MAPK signaling pathways as well as on the D. discoideum toll/interleukin-1 receptor domain-containing protein, TirA. These findings indicate that pattern-recognition machinery required to detect and respond to bacterial PAMPs may be conserved in D. discoideum. PMID:21527280

Walk, Alexander; Callahan, Jennifer; Srisawangvong, Pat; Leuschner, Jessica; Samaroo, Dave; Cassilly, Daniel; Snyder, Michelle L D

2011-04-19

275

Lipopolysaccharide enhances bactericidal activity in Dictyostelium discoideum cells.  

UK PubMed Central (United Kingdom)

Innate immune cells respond to invading microbes upon detection of pathogen-associated molecular patterns (PAMPS). PAMP-recognition machinery is evolutionarily conserved, allowing for characterization in model organisms. The model organism Dictyostelium discoideum can exist as single-celled amoebae, which phagocytize bacteria for nutrients. Although D. discoideum is used extensively to study phagocytosis, it has not been determined if D. discoideum detects bacterial PAMPs using pattern-recognition machinery. Here we show that D. discoideum mounts responses against the bacterial cell wall PAMP, lipopolysaccharide (LPS). Upon treatment with LPS or its active component Lipid A, D. discoideum cells more efficiently clear phagocytized bacteria. LPS-enhanced bactericidal activity appears dependent both on MAPK signaling pathways as well as on the D. discoideum toll/interleukin-1 receptor domain-containing protein, TirA. These findings indicate that pattern-recognition machinery required to detect and respond to bacterial PAMPs may be conserved in D. discoideum.

Walk A; Callahan J; Srisawangvong P; Leuschner J; Samaroo D; Cassilly D; Snyder ML

2011-08-01

276

E. coli Lipopolysaccharide: acute oral toxicity study in mice.  

UK PubMed Central (United Kingdom)

A single dose of endotoxin (lipopolysaccharide) from a common laboratory cloning and expression strain (Escherichia coli BL21[DE3]) was administered to groups of male and female CD-1 mice (n=5/group) at doses up to 1,000,000 endotoxin units (EU) per mouse by oral gavage. The mice were observed for mortality, body weight effects, and clinical signs for 14 days after which they were sacrificed for gross organ necropsy. All mice survived until the scheduled sacrifice, no clinical signs of toxicity were observed, no test substance-related body weight losses occurred and no gross lesions were present at necropsy. Under the conditions of this study, oral administration of E. coli BL21(DE3) endotoxin to mice at a dose of up to 1,000,000 EU/mouse produced no evidence of toxicity.

Harper MS; Carpenter C; Klocke DJ; Carlson G; Davis T; Delaney B

2011-08-01

277

Repeated injections of lipopolysaccharide attenuate the satiety effects of cholecystokinin.  

UK PubMed Central (United Kingdom)

The present study examined the effects of repeated injections of lipopolysaccharide (LPS) and cholecystokinin (CCK) on both sucrose palatability and body weight. Rats received repeated injections of LPS (200 microg/kg), CCK (8 microg/kg) and/or NaCl on days 1, 4, 7 and 10. Body weight was monitored on each test day and sucrose palatability was assessed using the taste reactivity test (TRT) on days 7 and 10. Rats treated with LPS developed a rapid tolerance to the reductions in body weight; however, this tolerance did not affect sucrose palatability. Furthermore it was found that repeated exposures to LPS and CCK attenuated the typical satiety related behaviors produced in the TRT by administration of CCK. This desensitization to CCK with repeated exposures to LPS may be one of the mechanisms that could account for the rapid development of tolerance to LPS-induced hypophagia.

Cross-Mellor SK; Kavaliers M; Ossenkopp KP

1999-12-01

278

Repeated injections of lipopolysaccharide attenuate the satiety effects of cholecystokinin.  

Science.gov (United States)

The present study examined the effects of repeated injections of lipopolysaccharide (LPS) and cholecystokinin (CCK) on both sucrose palatability and body weight. Rats received repeated injections of LPS (200 microg/kg), CCK (8 microg/kg) and/or NaCl on days 1, 4, 7 and 10. Body weight was monitored on each test day and sucrose palatability was assessed using the taste reactivity test (TRT) on days 7 and 10. Rats treated with LPS developed a rapid tolerance to the reductions in body weight; however, this tolerance did not affect sucrose palatability. Furthermore it was found that repeated exposures to LPS and CCK attenuated the typical satiety related behaviors produced in the TRT by administration of CCK. This desensitization to CCK with repeated exposures to LPS may be one of the mechanisms that could account for the rapid development of tolerance to LPS-induced hypophagia. PMID:10716221

Cross-Mellor, S K; Kavaliers, M; Ossenkopp, K P

1999-12-16

279

Potential role for lipopolysaccharide in congenital sensorineural hearing loss.  

Science.gov (United States)

Congenital sensorineural hearing loss (SNHL) is common. In the Western world, the incidence is 1-3 per 1000 live births. The aetiology encompasses genetic and non-genetic factors accounting for 55 % and 45 % of cases, respectively. Reports that describe the contribution of intrauterine infection to the occurrence of congenital SNHL are limited, and comparative analysis of the different pathogens is lacking. Lipopolysaccharide (LPS), a product of bacteriolysis, has been demonstrated to be associated with inner ear damage in experimental studies. To elucidate the potential role of this toxin in congenital SNHL and to identify the pathogenesis and transmission routes, we reviewed the literature. We speculate that different routes of exposure to LPS in utero may result in congenital inner ear damage. PMID:20093374

Smit, A L; Stokroos, R J; Litjens, S G H; Kremer, B; Kramer, B W

2010-01-21

280

Immunostimulants in teleost fish: probiotics, ?-glucans and lipopolysaccharides  

Directory of Open Access Journals (Sweden)

Full Text Available AbstractImmunostimulants mainly consist of microorganisms’ structural elements whose principle is based on stimulating the innate immune system, leading to an improvement in animals’ sanitary state and increased resistance to pathogens. ?-glucans and lipopolysaccharides are some of the most used immunostimulants in the fish industry, including beneficial bacteria called probiotics. Studies on fish have been focused on the in vitro and in vivo evaluation of cellular and humoral responses, modulating gene transcription and the effects of resistance regarding pathogens of interest, usually having positive effects on the fishes’ immunological state and resistance to disease. The proposed review was aimed at investigating the mechanisms of action of some of the most used immunostimulants, conceptualising their use in aquiculture and discussing recent work on the topic thereby forming the basis for proposing pertinent, feasible investigations for tackling some problems in the fish-farming health/sanitation area.

Mónica A. Vásquez - Piñeros; Iang S. Rondón - Barragan; Pedro R. Eslava- Mocha

2012-01-01

 
 
 
 
281

Potential role for lipopolysaccharide in congenital sensorineural hearing loss.  

UK PubMed Central (United Kingdom)

Congenital sensorineural hearing loss (SNHL) is common. In the Western world, the incidence is 1-3 per 1000 live births. The aetiology encompasses genetic and non-genetic factors accounting for 55 % and 45 % of cases, respectively. Reports that describe the contribution of intrauterine infection to the occurrence of congenital SNHL are limited, and comparative analysis of the different pathogens is lacking. Lipopolysaccharide (LPS), a product of bacteriolysis, has been demonstrated to be associated with inner ear damage in experimental studies. To elucidate the potential role of this toxin in congenital SNHL and to identify the pathogenesis and transmission routes, we reviewed the literature. We speculate that different routes of exposure to LPS in utero may result in congenital inner ear damage.

Smit AL; Stokroos RJ; Litjens SG; Kremer B; Kramer BW

2010-04-01

282

Autophagy in periodontitis patients and gingival fibroblasts: unraveling the link between chronic diseases and inflammation  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Periodontitis, the most prevalent chronic inflammatory disease, has been related to cardiovascular diseases. Autophagy provides a mechanism for the turnover of cellular organelles and proteins through a lysosome-dependent degradation pathway. The aim of this research was to study the role of autophagy in peripheral blood mononuclear cells from patients with periodontitis and gingival fibroblasts treated with a lipopolysaccharide of Porphyromonas gingivalis. Autophagy-dependent mechanisms have been proposed in the pathogenesis of inflammatory disorders and in other diseases related to periodontitis, such as cardiovascular disease and diabetes. Thus it is important to study the role of autophagy in the pathophysiology of periodontitis. Methods Peripheral blood mononuclear cells from patients with periodontitis (n = 38) and without periodontitis (n = 20) were used to study autophagy. To investigate the mechanism of autophagy, we evaluated the influence of a lipopolysaccharide from P. gingivalis in human gingival fibroblasts, and autophagy was monitored morphologically and biochemically. Autophagosomes were observed by immunofluorescence and electron microscopy. Results We found increased levels of autophagy gene expression and high levels of mitochondrial reactive oxygen species production in peripheral blood mononuclear cells from patients with periodontitis compared with controls. A significantly positive correlation between both was observed. In human gingival fibroblasts treated with lipopolysaccharide from P. gingivalis, there was an increase of protein and transcript of autophagy-related protein 12 (ATG12) and microtubule-associated protein 1 light chain 3 alpha LC3. A reduction of mitochondrial reactive oxygen species induced a decrease in autophagy whereas inhibition of autophagy in infected cells increased apoptosis, showing the protective role of autophagy. Conclusion Results from the present study suggest that autophagy is an important and shared mechanism in other conditions related to inflammation or alterations of the immune system, such as periodontitis.

Bullon Pedro; Cordero Mario; Quiles José; Ramirez-Tortosa Maria; Gonzalez-Alonso Adrian; Alfonsi Simona; García-Marín Rocio; de Miguel Manuel; Battino Maurizio

2012-01-01

283

Characterization and functional study of Antrodia camphorata lipopolysaccharide.  

UK PubMed Central (United Kingdom)

Lipopolysaccharide (LPS) is a highly proinflammatory molecule isolated from bacteria. This study demonstrated the existence of LPS in a medicinal fungus, Antrodia camphorata. Because no LPS had been identified in any fungus organism, the purification of LPS from A. camphorata was attempted. LPSs from six strains of A. camphorata (35396, 35398, 35716, B71, B85, and B86) were isolated. Chemical and functional properties were investigated on the fungus LPS. Compositional analysis revealed that sorbitol, fucose, galactose, and glucose were the neutral sugars in LPS of A. camphorata. Galactosamine, glucosamine, galactose, and glucose were the predominant monosaccharide species in E. coli O129 LPS molecules, whereas galactosamine and glucosamine were absent in A. camphorata LPS. Because these properties are different from those of bacterial LPS, the functions between fungus and bacterial LPS are also discussed. The vascular endothelial lining of blood vessels, which controls leucocyte traffic and activation, may be one of the primary targets of LPS action during sepsis. Assays for biological activity were performed on endothelial cells with anti-inflammatory effects associated with sepsis. A. camphorata LPS apparently showed a lesser extent of cytotoxicity than bacterial LPS. In contrary to the proinflammatory property of bacterial LPS, LPS from A. camphorata differentially reversed bacterial LPS-induced intercellular adhersion molecule-1 and monocyte adhesion; both were indicators during inflammatory process. In conclusion, basic chemical properties categorized A. camphorata extracts into lipopolysaccharide. However, the detailed functional structures and bioactivities of A. camphorata LPS were totally different from those of bacterial LPS. The investigation of the existence and anti-inflammatory effect of fungus LPS is at present a truly novel and important finding. These results show that LPS isolated from A. camphorata offers a novel therapeutic target for anti-inflammation against E. coli infection.

Cheng JJ; Yang CJ; Cheng CH; Wang YT; Huang NK; Lu MK

2005-01-01

284

Direct evidence for the presence of lipopolysaccharide components in Pseudomonas ribosomal vaccine.  

UK PubMed Central (United Kingdom)

The presence of sugars specific to lipopolysaccharide, glucose, and rhamnose was demonstrated in a Pseudomonas ribosomal vaccine. The detection of these sugars was accomplished by radiological means after paper chromatography of the neutral fraction of acid-hydrolyzed vaccine.

Lieberman MM

1977-08-01

285

Structural Analysis and Involvement in Plant Innate Immunity of Xanthomonas axonopodis pv. citri Lipopolysaccharide*  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Xanthomonas axonopodis pv. citri (Xac) causes citrus canker, provoking defoliation and premature fruit drop with concomitant economical damage. In plant pathogenic bacteria, lipopolysaccharides are important virulence factors, and they are being increasingly recognized as major pathogen-associated m...

Casabuono, Adriana; Petrocelli, Silvana; Ottado, Jorgelina; Orellano, Elena G.; Couto, Alicia S.

286

Clindamycin Modulates Inflammatory-Cytokine Induction in Lipopolysaccharide-Stimulated Mouse Peritoneal Macrophages  

Digital Repository Infrastructure Vision for European Research (DRIVER)

We investigated the mechanism by which clindamycin (CLI) modulates cytokine induction after lipopolysaccharide (LPS) stimulation. Although CLI decreased the intracellular expression levels of tumor necrosis factor alpha and interleukin 1? (IL-1?) and increased IL-6 expression in macrophages, cytokin...

Nakano, Tetsuji; Hiramatsu, Kazufumi; Kishi, Kenji; Hirata, Norio; Kadota, Jun-ichi; Nasu, Masaru

287

Modification of biological activity of lipopolysaccharide in the complex with chitosan.  

Science.gov (United States)

In the complex with chitosan, lipopolysaccharide partially lost its ability to induce lymphokines tumor necrosis factor and interleukin-8, but retained immunostimulating properties and increased phagocytic function of macrophages by improving digestion of bacteria. PMID:15452608

Ermak, I M; Davydova, V N; Gorbach, V I; Berdyshev, E L; Kuznetsova, T A; Ivanushko, I A; Gazha, A K; Smolina, T P; Zaporozhets, T S; Solov'eva, T F

2004-04-01

288

Selective sorting of cargo proteins into bacterial membrane vesicles.  

UK PubMed Central (United Kingdom)

In contrast to the well established multiple cellular roles of membrane vesicles in eukaryotic cell biology, outer membrane vesicles (OMV) produced via blebbing of prokaryotic membranes have frequently been regarded as cell debris or microscopy artifacts. Increasingly, however, bacterial membrane vesicles are thought to play a role in microbial virulence, although it remains to be determined whether OMV result from a directed process or from passive disintegration of the outer membrane. Here we establish that the human oral pathogen Porphyromonas gingivalis has a mechanism to selectively sort proteins into OMV, resulting in the preferential packaging of virulence factors into OMV and the exclusion of abundant outer membrane proteins from the protein cargo. Furthermore, we show a critical role for lipopolysaccharide in directing this sorting mechanism. The existence of a process to package specific virulence factors into OMV may significantly alter our current understanding of host-pathogen interactions.

Haurat MF; Aduse-Opoku J; Rangarajan M; Dorobantu L; Gray MR; Curtis MA; Feldman MF

2011-01-01

289

Selective sorting of cargo proteins into bacterial membrane vesicles.  

Science.gov (United States)

In contrast to the well established multiple cellular roles of membrane vesicles in eukaryotic cell biology, outer membrane vesicles (OMV) produced via blebbing of prokaryotic membranes have frequently been regarded as cell debris or microscopy artifacts. Increasingly, however, bacterial membrane vesicles are thought to play a role in microbial virulence, although it remains to be determined whether OMV result from a directed process or from passive disintegration of the outer membrane. Here we establish that the human oral pathogen Porphyromonas gingivalis has a mechanism to selectively sort proteins into OMV, resulting in the preferential packaging of virulence factors into OMV and the exclusion of abundant outer membrane proteins from the protein cargo. Furthermore, we show a critical role for lipopolysaccharide in directing this sorting mechanism. The existence of a process to package specific virulence factors into OMV may significantly alter our current understanding of host-pathogen interactions. PMID:21056982

Haurat, M Florencia; Aduse-Opoku, Joseph; Rangarajan, Minnie; Dorobantu, Loredana; Gray, Murray R; Curtis, Michael A; Feldman, Mario F

2010-11-05

290

Structural Analysis and Involvement in Plant Innate Immunity of Xanthomonas axonopodis pv. citri Lipopolysaccharide*  

Science.gov (United States)

Xanthomonas axonopodis pv. citri (Xac) causes citrus canker, provoking defoliation and premature fruit drop with concomitant economical damage. In plant pathogenic bacteria, lipopolysaccharides are important virulence factors, and they are being increasingly recognized as major pathogen-associated molecular patterns for plants. In general, three domains are recognized in a lipopolysaccharide: the hydrophobic lipid A, the hydrophilic O-antigen polysaccharide, and the core oligosaccharide, connecting lipid A and O-antigen. In this work, we have determined the structure of purified lipopolysaccharides obtained from Xanthomonas axonopodis pv. citri wild type and a mutant of the O-antigen ABC transporter encoded by the wzt gene. High pH anion exchange chromatography and matrix-assisted laser desorption/ionization mass spectrum analysis were performed, enabling determination of the structure not only of the released oligosaccharides and lipid A moieties but also the intact lipopolysaccharides. The results demonstrate that Xac wild type and Xacwzt LPSs are composed mainly of a penta- or tetra-acylated diglucosamine backbone attached to either two pyrophosphorylethanolamine groups or to one pyrophosphorylethanolamine group and one phosphorylethanolamine group. The core region consists of a branched oligosaccharide formed by Kdo2Hex6GalA3Fuc3NAcRha4 and two phosphate groups. As expected, the presence of a rhamnose homo-oligosaccharide as O-antigen was determined only in the Xac wild type lipopolysaccharide. In addition, we have examined how lipopolysaccharides from Xac function in the pathogenesis process. We analyzed the response of the different lipopolysaccharides during the stomata aperture closure cycle, the callose deposition, the expression of defense-related genes, and reactive oxygen species production in citrus leaves, suggesting a functional role of the O-antigen from Xac lipopolysaccharides in the basal response.

Casabuono, Adriana; Petrocelli, Silvana; Ottado, Jorgelina; Orellano, Elena G.; Couto, Alicia S.

2011-01-01

291

Structural analysis and involvement in plant innate immunity of Xanthomonas axonopodis pv. citri lipopolysaccharide.  

UK PubMed Central (United Kingdom)

Xanthomonas axonopodis pv. citri (Xac) causes citrus canker, provoking defoliation and premature fruit drop with concomitant economical damage. In plant pathogenic bacteria, lipopolysaccharides are important virulence factors, and they are being increasingly recognized as major pathogen-associated molecular patterns for plants. In general, three domains are recognized in a lipopolysaccharide: the hydrophobic lipid A, the hydrophilic O-antigen polysaccharide, and the core oligosaccharide, connecting lipid A and O-antigen. In this work, we have determined the structure of purified lipopolysaccharides obtained from Xanthomonas axonopodis pv. citri wild type and a mutant of the O-antigen ABC transporter encoded by the wzt gene. High pH anion exchange chromatography and matrix-assisted laser desorption/ionization mass spectrum analysis were performed, enabling determination of the structure not only of the released oligosaccharides and lipid A moieties but also the intact lipopolysaccharides. The results demonstrate that Xac wild type and Xacwzt LPSs are composed mainly of a penta- or tetra-acylated diglucosamine backbone attached to either two pyrophosphorylethanolamine groups or to one pyrophosphorylethanolamine group and one phosphorylethanolamine group. The core region consists of a branched oligosaccharide formed by Kdo?Hex?GalA?Fuc3NAcRha? and two phosphate groups. As expected, the presence of a rhamnose homo-oligosaccharide as O-antigen was determined only in the Xac wild type lipopolysaccharide. In addition, we have examined how lipopolysaccharides from Xac function in the pathogenesis process. We analyzed the response of the different lipopolysaccharides during the stomata aperture closure cycle, the callose deposition, the expression of defense-related genes, and reactive oxygen species production in citrus leaves, suggesting a functional role of the O-antigen from Xac lipopolysaccharides in the basal response.

Casabuono A; Petrocelli S; Ottado J; Orellano EG; Couto AS

2011-07-01

292

Isoforskolin pretreatment attenuates lipopolysaccharide-induced acute lung injury in animal models.  

Science.gov (United States)

Isoforskolin was isolated from Coleus forskohlii native to Yunnan in China. We hypothesize that isoforskolin pretreatment attenuates acute lung injury induced by lipopolysaccharide (endotoxin). Three acute lung injury models were used: situ perfused rat lung, rat and mouse models of endotoxic shock. Additionally, lipopolysaccharide stimulated proinflammatory cytokine production was evaluated in human mononuclear leukocyte. In situ perfused rat lungs, pre-perfusion with isoforskolin (100, and 200 ?M) and dexamethasone (65 ?M, positive control) inhibited lipopolysaccharide (10 mg/L) induced increases in lung neutrophil adhesion rate, myeloperoxidase activity, lung weight Wet/Dry ratio, permeability-surface area product value, and tumor necrosis factor (TNF)-? levels. In rats, pretreatments with isoforskolin (5, 10, and 20 mg/kg, i.p.) and dexamethasone (5mg/kg, i.p.) markedly reduced lipopolysaccharide (6 mg/kg i.v.) induced increases of karyocyte, neutrophil counts and protein content in bronchoalveolar lavage fluid, and plasma myeloperoxidase activity. Lung histopathology showed that morphologic changes induced by lipopolysaccharide were less pronounced in the isoforskolin and dexamethasone pretreated rats. In mice, 5 mg/kg isoforskolin and dexamethasone caused 100% and 80% survival, respectively, after administration of lipopolysaccharide (62.5mg/kg, i.v., 40% survival if untreated). In human mononuclear leukocyte, isoforskolin (50, 100, and 200 ?M) and dexamethasone (10 ?M) pre-incubation lowered lipopolysaccharide (2 ?g/mL) induced secretion of the cytokine TNF-?, and interleukins (IL)-1?, IL-6, and IL-8. In conclusion, pretreatment with isoforskolin attenuates lipopolysaccharide-induced acute lung injury in several models, and it is involved in down-regulation of inflammatory responses and proinflammatory cytokines TNF-?, IL-1?, IL-6, and IL-8. PMID:21272678

Yang, Weimin; Qiang, Dongjin; Zhang, Min; Ma, Limei; Zhang, Yonghui; Qing, Chen; Xu, Yunlong; Zhen, Chunlan; Liu, Jikai; Chen, Yan-Hua

2011-01-25

293

Immunization of dairy cows with an Escherichia coli J5 lipopolysaccharide vaccine.  

UK PubMed Central (United Kingdom)

Development of a lipopolysaccharide-protein conjugate vaccine and the immunological response to the vaccine were investigated. Lipopolysaccharide derived from Escherichia coli J5 was detoxified by mild alkaline hydrolysis. Detoxification reduced endotoxin activity 2500-fold compared with that of native J5 lipopolysaccharide. The conjugate vaccine was synthesized by covalently coupling detoxified lipopolysaccharide to chicken serum albumin by reductive amination. Dairy cows were immunized with 8.35 mg of conjugate (n = 3) or 5 x 10(9) heat-killed J5 bacterin (n = 5) at 215 DIM and received a secondary immunization 14 d later. Control cows were not immunized. Immunization enhanced serum antibody titer to J5 lipopolysaccharide antigens. Whey IgG and IgM titers to J5 lipopolysaccharide were not different among treatment groups. Serum and whey IgG titers to J5 whole-cell antigens were elevated in immunized cows within treatment groups. Immunization did not enhance whey IgM to J5 whole-cell antigens. Conjugate immunization elicited an immune response comparable with or greater than that of immunized cows with J5 bacterin.

Tomita GM; Todhunter DA; Hogan JS; Smith KL

1995-10-01

294

Bitter gourd suppresses lipopolysaccharide-induced inflammatory responses.  

Science.gov (United States)

Bitter gourd ( Momordica charantia L.) is a popular tropical vegetable in Asian countries. Previously it was shown that bitter gourd placenta extract suppressed lipopolysaccharide (LPS)-induced TNFalpha production in RAW 264.7 macrophage-like cells. Here it is shown that the butanol-soluble fraction of bitter gourd placenta extract strongly suppresses LPS-induced TNFalpha production in RAW 264.7 cells. Gene expression analysis using a fibrous DNA microarray showed that the bitter gourd butanol fraction suppressed expression of various LPS-induced inflammatory genes, such as those for TNF, IL1alpha, IL1beta, G1p2, and Ccl5. The butanol fraction significantly suppressed NFkappaB DNA binding activity and phosphorylation of p38, JNK, and ERK MAPKs. Components in the active fraction from bitter gourd were identified as 1-alpha-linolenoyl-lysophosphatidylcholine (LPC), 2-alpha-linolenoyl-LPC, 1-lynoleoyl-LPC, and 2-linoleoyl-LPC. Purified 1-alpha-linolenoyl-LPC and 1-linoleoyl-LPC suppressed the LPS-induced TNFalpha production of RAW 264.7 cells at a concentration of 10 microg/mL. PMID:18489106

Kobori, Masuko; Nakayama, Hirosuke; Fukushima, Kenji; Ohnishi-Kameyama, Mayumi; Ono, Hiroshi; Fukushima, Tatsunobu; Akimoto, Yukari; Masumoto, Saeko; Yukizaki, Chizuko; Hoshi, Yoshikazu; Deguchi, Tomoaki; Yoshida, Mitsuru

2008-05-20

295

Lipopolysaccharides induces MUC5AC overproduction in human nasal epithelium.  

UK PubMed Central (United Kingdom)

Hyperproduction of mucin in the nasal epithelium is an important feature of nasal inflammatory diseases. We investigated the mechanism of lipopolysaccharides (LPS) involvement in mucin 5 subtype AC (MUC5AC) expression in human nasal epithelial cells. The primary human nasal epithelial cells were cultured in vitro, which were treated with LPS (10 nM/ml or 1 ?M/ml) for 12 and 24 h. LPS-induced MUC5AC protein was determined in nasal epithelial cells. The levels of nuclear factor kappa B p65 (NF-?Bp65) and its inhibitor kappa B? (I?B?) protein were also detected, and interleukin-1? (IL-1?) mRNA was detected by real-time PCR. LPS up-regulated MUC5AC protein in human nasal epithelial cells, and we determined that the up-regulation of MUC5AC expression was due to a time- and dose-dependent degradation of I?B? protein, which resulted in the increase of NF-?Bp65 nuclear translocation. Subsequently, we also determined that LPS can induce IL-1? mRNA in a time- and dose-dependent manner. These data show that LPS treatment activated NF-?B by promoting the degradation of I?B? and the nuclear localization of NF-?Bp65, which induced MUC5AC overproduction.

Wang W; Xu X; Zheng M; Wan L

2013-02-01

296

Biological activities of lipopolysaccharide fractionated by preparative acrylamide gel electrophoresis.  

UK PubMed Central (United Kingdom)

Lipopolysaccharide from a smooth strain of Salmonella minnesota was fractionated into two major fractions and one intermediate fraction by using sodium dodecylsulfate-polyacrylamide gel electrophoresis. On the basis of the study by Hitchcock and Brown, it was deduced that the top fraction was mainly long O-side chain LPS and the bottom fraction was O-side chain-less LPS. The middle fraction was a mixture of both short O-side chain LPS and O-side chain-less LPS. The antigenic properties and biological activities were not altered in this fractionation procedure. Comparison of the biological activities of the top fraction with those of the bottom fraction revealed that the bottom fraction had higher activity in polyclonal B-cell activation and spleen-swelling effect and that there was no significant difference in adjuvant activity, ability to render macrophages cytotoxic, induction of colony-stimulating factor and the ability to induce the Schwartzmann reaction. It was suggested that O-side chain makes no contribution to the latter biological activities including adjuvant activity of S. minnesota LPS.

Ohta M; Rothmann J; Kovats E; Pham PH; Nowotny A

1985-01-01

297

Wip1 Phosphatase Involved in Lipopolysaccharide-Induced Neuroinflammation.  

UK PubMed Central (United Kingdom)

Wild type p53-induced phosphatase 1 (Wip1) is a phosphatase which belongs to protein phosphatase type 2C family, which have been predominantly linked to cell growth and to cellular stress signaling. Numerous downstream targets of Wip1 have been identified, and genetic studies confirm that some play a part in tumorigenesis. Recent evidence highlights a new role for Wip1 in the regulation of NF-?B p65, which indicated that it might play a critical role in immune system. However, its regulation role in central nervous system (CNS) remains poorly understood. To elaborate whether Wip1 was involved in CNS injury, we performed a neuroinflammatory model by lipopolysaccharide (LPS) lateral-ventral injection in adult rats. Wip1 expression was strongly upregulated in active astrocytes in inflamed brain cortex. In vitro studies indicated that the upregulation of Wip1 may be involved in the subsequent astrocytic activation following LPS exposure, and knockdown of Wip1 in primary astrocytes by siRNA showed that Wip1 inhibited the synthesis of TNF-?. Collectively, these results suggested that Wip1 may be important in host defense in CNS immune response, which might provide a potent therapeutic target of neuroinflammation.

Tan X; Zhang J; Jin W; Li L; Xu W; Zheng H; Rui Y; Ke K; Zhou R; Cao M; Pan Y

2013-08-01

298

Bacterial lipopolysaccharide protects the retina from light-induced damage.  

Science.gov (United States)

Light-induced damage is a widely used model to study retinal degeneration. We examined whether bacterial lipopolysaccharide (LPS) protects the retina against light-induced injury. One day before intense light exposure for 24 h, rats were intravitreally injected with LPS in one eye and vehicle in the contralateral eye. At several time points after light exposure, rats were subjected to electroretinography and histological analysis. Bax, Bcl-xL, p-Akt, and p-Stat3 levels were assessed by Western blotting, and retinal thiobarbituric acid reactive substances levels were measured as an index of lipid peroxidation. One group of animals received injections of dexamethasone, aminoguanidine (an inducible NOS inhibitor), 5-hydroxydecanoic acid (a mitochondrial K(+) /ATP channel blocker), or wortmannin [a phosphoinositide-3-kinase (PI3K) inhibitor] in order to analyze their effect on the protection induced by LPS. LPS afforded significant morphologic and functional protection in eyes exposed to intense light. Light damage induced an increase in mitochondrial Bax/cytoplasmic Bax ratio, and lipid peroxidation which were prevented by LPS. Dexamethasone and wortmannin (but not aminoguanidine or 5-hydroxydecanoic acid) prevented the effect of LPS. Moreover, wortmannin prevented the effect of LPS on p-Akt levels. These results indicate that LPS provides retinal protection against light-induced stress, probably through a PI3K/Akt-dependent mechanism. PMID:22536982

Bordone, Melina P; Lanzani, María Florencia; López-Costa, Juan José; Chianelli, Mónica S; Franco, Pablo; Sáenz, Daniel A; Rosenstein, Ruth E

2012-05-23

299

Bacterial lipopolysaccharide protects the retina from light-induced damage.  

UK PubMed Central (United Kingdom)

Light-induced damage is a widely used model to study retinal degeneration. We examined whether bacterial lipopolysaccharide (LPS) protects the retina against light-induced injury. One day before intense light exposure for 24 h, rats were intravitreally injected with LPS in one eye and vehicle in the contralateral eye. At several time points after light exposure, rats were subjected to electroretinography and histological analysis. Bax, Bcl-xL, p-Akt, and p-Stat3 levels were assessed by Western blotting, and retinal thiobarbituric acid reactive substances levels were measured as an index of lipid peroxidation. One group of animals received injections of dexamethasone, aminoguanidine (an inducible NOS inhibitor), 5-hydroxydecanoic acid (a mitochondrial K(+) /ATP channel blocker), or wortmannin [a phosphoinositide-3-kinase (PI3K) inhibitor] in order to analyze their effect on the protection induced by LPS. LPS afforded significant morphologic and functional protection in eyes exposed to intense light. Light damage induced an increase in mitochondrial Bax/cytoplasmic Bax ratio, and lipid peroxidation which were prevented by LPS. Dexamethasone and wortmannin (but not aminoguanidine or 5-hydroxydecanoic acid) prevented the effect of LPS. Moreover, wortmannin prevented the effect of LPS on p-Akt levels. These results indicate that LPS provides retinal protection against light-induced stress, probably through a PI3K/Akt-dependent mechanism.

Bordone MP; Lanzani MF; López-Costa JJ; Chianelli MS; Franco P; Sáenz DA; Rosenstein RE

2012-07-01

300

Zymosan: induction of sickness behavior and interaction with lipopolysaccharide.  

Science.gov (United States)

The yeast particulate zymosan (Zy) activates innate immune system cells and induces cytokine secretion. There is also evidence that Zy can affect biologic responses to bacterial lipopolysaccharide (LPS) and that the pathways by which these two agents act upon immune cells are only partially distinct. The present experiments assessed the ability of Zy to elicit CNS-mediated sickness symptoms and to alter their responses to LPS. In Experiment 1, Zy induced elements of the sickness behavior syndrome dose-responsively in Long-Evans rats, as indicated by reductions in consumption of a highly palatable bait and in body temperature. In Experiment 2, Zy exerted a priming effect, sensitizing animals to subsequent LPS as measured by reductions in bait consumption, 24-h laboratory chow intake, and body temperature. Experiment 3 failed to provide evidence for LPS-to-Zy cross-tolerance but did indicate that the administration of Zy disrupts previously acquired LPS tolerance. These results suggest that the specifics of exposure to microbially derived innate immune activators have to be taken into account in investigating the biologic bases of sickness behaviors and developing models of coinfection. PMID:14637214

Cremeans-Smith, Julie K; Newberry, Benjamin H

2003-11-01

 
 
 
 
301

Role of nitric oxide in tolerance to lipopolysaccharide in mice.  

UK PubMed Central (United Kingdom)

The injection of repeated doses of lipopolysaccharide (LPS) results in attenuation of the febrile response, which is called endotoxin tolerance. We tested the hypothesis that nitric oxide (NO) arising from inducible NO synthase (iNOS) plays a role in endotoxin tolerance, using not only pharmacological trials but also genetically engineered mice. Body core temperature was measured by biotelemetry in mice treated with NG-monomethyl-L-arginine (L-NMMA, 40 mg/kg; a nonselective NO synthase inhibitor) or aminoguanidine (AG, 10 mg/kg; a selective iNOS inhibitor) and in mice deficient in the iNOS gene (iNOS KO) mice. Tolerance to LPS was induced by means of three consecutive LPS (100 microg/kg) intraperitoneal injections at 24-h intervals. In wild-type mice, we observed a significant reduction of the febrile response to repeated administration of LPS. Injection of L-NMMA and AG markedly enhanced the febrile response to LPS in tolerant animals. Conversely, iNOS-KO mice repeatedly injected with LPS did not become tolerant to the pyrogenic effect of LPS. These data are consistent with the notion that NO modulates LPS tolerance in mice and that iNOS isoform is involved in NO synthesis during LPS tolerance.

Dias MB; Almeida MC; Carnio EC; Branco LG

2005-04-01

302

Role of nitric oxide in tolerance to lipopolysaccharide in mice.  

Science.gov (United States)

The injection of repeated doses of lipopolysaccharide (LPS) results in attenuation of the febrile response, which is called endotoxin tolerance. We tested the hypothesis that nitric oxide (NO) arising from inducible NO synthase (iNOS) plays a role in endotoxin tolerance, using not only pharmacological trials but also genetically engineered mice. Body core temperature was measured by biotelemetry in mice treated with NG-monomethyl-L-arginine (L-NMMA, 40 mg/kg; a nonselective NO synthase inhibitor) or aminoguanidine (AG, 10 mg/kg; a selective iNOS inhibitor) and in mice deficient in the iNOS gene (iNOS KO) mice. Tolerance to LPS was induced by means of three consecutive LPS (100 microg/kg) intraperitoneal injections at 24-h intervals. In wild-type mice, we observed a significant reduction of the febrile response to repeated administration of LPS. Injection of L-NMMA and AG markedly enhanced the febrile response to LPS in tolerant animals. Conversely, iNOS-KO mice repeatedly injected with LPS did not become tolerant to the pyrogenic effect of LPS. These data are consistent with the notion that NO modulates LPS tolerance in mice and that iNOS isoform is involved in NO synthesis during LPS tolerance. PMID:15579566

Dias, Mirela B; Almeida, Maria C; Carnio, Evelin C; Branco, Luiz G S

2004-12-03

303

Lipopolysaccharide induces autotaxin expression in human monocytic THP-1 cells  

International Nuclear Information System (INIS)

[en] Autotaxin (ATX) is a secreted enzyme with lysophospholipase D (lysoPLD) activity, which converts lysophosphatidylcholine (LPC) into lysophosphatidic acid (LPA), a bioactive phospholipid involved in numerous biological activities, including cell proliferation, differentiation, and migration. In the present study, we found that bacterial lipopolysaccharide (LPS), a well-known initiator of the inflammatory response, induced ATX expression in monocytic THP-1 cells. The activation of PKR, JNK, and p38 MAPK was required for the ATX induction. The LPS-induced ATX in THP-1 cells was characterized as the ? isoform. In the presence of LPC, ATX could promote the migrations of THP-1 and Jurkat cells, which was inhibited by pertussis toxin (PTX), an inhibitor of Gi-mediated LPA receptor signaling. In summary, LPS induces ATX expression in THP-1 cells via a PKR, JNK and p38 MAPK-mediated mechanism, and the ATX induction is likely to enhance immune cell migration in proinflammatory response by regulating LPA levels in the microenvironment.

2009-01-09

304

Evaluation of lipopolysaccharide aggregation by light scattering spectroscopy.  

Science.gov (United States)

Lipopolysaccharides (LPS) are cell wall components of Gram-negative bacteria. These molecules behave as bacterial endotoxins and their release into the bloodstream is a determinant of the development of a wide range of pathologies. These amphipathic molecules can self-aggregate into supramolecular structures with different shapes and sizes. The formation of these structures occurs when the LPS concentration is higher than the apparent critical micelle concentration (CMC(a)). Light scattering spectroscopy (both static and dynamic) was used to directly characterize the aggregation process of LPS from Escherichia coli serotype 026:B6. The results point to a CMC(a) value of 14 microg mL(-1) and the existence of premicelle LPS oligomers below this concentration. Both structures were characterized in terms of molecular weight (5.5 x 10(6) and 16 x 10(6) g mol(-1) below and above the CMC(a), respectively), interaction with the aqueous environment, gyration radius (56 and 105 nm), hydrodynamic radius, (60 and 95 nm) and geometry of the supramolecular structures (nearly spherical). Our data indicates that future in vitro experiments should be carried out both below and above the CMC(a). The search for drugs that interact with the aggregates, and thus change the CMC(a) and condition LPS interactions in the bloodstream, could be a new way to prevent certain bacterial-endotoxin-related pathologies. PMID:12512082

Santos, Nuno C; Silva, Ana C; Castanho, Miguel A R B; Martins-Silva, J; Saldanha, Carlota

2003-01-01

305

Evaluation of lipopolysaccharide aggregation by light scattering spectroscopy.  

UK PubMed Central (United Kingdom)

Lipopolysaccharides (LPS) are cell wall components of Gram-negative bacteria. These molecules behave as bacterial endotoxins and their release into the bloodstream is a determinant of the development of a wide range of pathologies. These amphipathic molecules can self-aggregate into supramolecular structures with different shapes and sizes. The formation of these structures occurs when the LPS concentration is higher than the apparent critical micelle concentration (CMC(a)). Light scattering spectroscopy (both static and dynamic) was used to directly characterize the aggregation process of LPS from Escherichia coli serotype 026:B6. The results point to a CMC(a) value of 14 microg mL(-1) and the existence of premicelle LPS oligomers below this concentration. Both structures were characterized in terms of molecular weight (5.5 x 10(6) and 16 x 10(6) g mol(-1) below and above the CMC(a), respectively), interaction with the aqueous environment, gyration radius (56 and 105 nm), hydrodynamic radius, (60 and 95 nm) and geometry of the supramolecular structures (nearly spherical). Our data indicates that future in vitro experiments should be carried out both below and above the CMC(a). The search for drugs that interact with the aggregates, and thus change the CMC(a) and condition LPS interactions in the bloodstream, could be a new way to prevent certain bacterial-endotoxin-related pathologies.

Santos NC; Silva AC; Castanho MA; Martins-Silva J; Saldanha C

2003-01-01

306

Effects of lipopolysaccharide on the catabolic activity of macrophages  

Energy Technology Data Exchange (ETDEWEB)

The ability of macrophages to degrade and catabolize antigens is of relevance both as a means to process complex antigens prior to presentation to T cells, as well as a way to down regulate immune responses by destroying the antigenicity of polypeptides. With these considerations, the authors have investigated the regulation of macrophage catabolic activity by lipopolysaccharide (LPS). Catabolic activity was quantitated by following the distribution and molecular form of /sup 125/-I labelled surface components of heat-killed Listeria monocytogenes (HKLM) subsequent to their uptake by macrophages. They have compared the catabolic activity of macrophages from peritoneal exudates of mice injected i.p. with saline or LPS and have found that LPS-elicited macrophages display a greatly enhanced (3 fold) rate of catabolism. This increase in catabolic activity peaks 3 days after LPS injection and steadily declines thereafter, approaching a baseline level after 3 weeks. The enhancement of catabolic activity is under LPS gene control. LPS-elicited macrophages rapidly destroy the antigenicity of bacterial antigens and function poorly as antigen presenting cells in vitro. These results suggest that LPS elicits a macrophage population specialized for antigen degradation functions with negative regulatory effects on the induction of specific immune responses.

Cluff, C.; Ziegler, H.K.

1986-03-05

307

Remifentanil attenuates human neutrophils activation induced by lipopolysaccharide.  

UK PubMed Central (United Kingdom)

The purpose of the present study was to investigate the effect of various opioids including remifentanil, sufentanil, alfentanil and fentanyl on human neutrophil activation induced by lipopolysaccharide (LPS) and to reveal the correlation involving the activation of cytokines and mitogen-activated protein kinases (MAPKs). Neutrophils from human blood were incubated with various concentrations of opioids with LPS. We measured protein levels for tumor necrosis factor-alpha (TNF-?), interleukin (IL)-6 and IL-8 after 4?h incubation period. To clarify the intracellular signaling pathway, we measured the levels of phosphorylation of p38, extracellular signal-regulated protein kinases 1 and 2 (ERK1/2) and c-Jun N-terminal kinase. We also measured the levels for cytokines and MAPKs to reveal the effects of several opioid receptor subtype antagonists on remifentanil with LPS. In the present experiment, only remifentanil could attenuate activation of human neutrophils exposed to LPS. In particular, remifentanil decreased activation of intracellular signaling pathways, including p38 and ERK1/2, and expression of pro-inflammatory cytokines, including TNF-?, IL-6 and IL-8. Especially the decreased activation of cytokines and MAPKs was significantly reverted by a kappa-opioid receptor antagonist on remifentanil with LPS. These results demonstrate that remifentanil can attenuate human neutrophils activations induced by LPS and a kappa-opioid receptor be probably involved in these anti-inflammatory effects mediated by remifentanil.

Hyejin J; Mei L; Seongheon L; Cheolwon J; Seokjai K; Hongbeom B; Minsun K; Sungsu C; Sanghyun K

2013-04-01

308

Intrabronchial activated protein C enhances lipopolysaccharide-induced pulmonary responses.  

UK PubMed Central (United Kingdom)

Intravenous administration of activated protein C (APC) inhibits coagulation and inflammation in the lungs of humans and animals. Investigations in rodents demonstrated that direct intrapulmonary delivery of APC also exerts anticoagulant and anti-inflammatory effects. The effect of intrabronchial administration of recombinant human (rh)APC on lipopolysaccharide (LPS)-induced haemostatic and inflammatory alterations in the bronchoalveolar space of humans was studied. Eight subjects received rhAPC via intrabronchial instillation by bronchoscope, while in a contralateral subsegment subjects received saline; all subjects were challenged bilaterally with LPS in the same lung subsegments. Four additional subjects received rhAPC (75 ?g), with saline as a control in the contralateral subsegment, while they were bilaterally "challenged" with saline. After 6 h a bronchoalveolar lavage was performed and coagulation and inflammatory parameters were measured. rhAPC enhanced LPS-induced coagulation activation in the bronchoalveolar space, when compared with the control side. In addition, rhAPC amplified LPS-induced pro-inflammatory responses, as indicated by higher concentrations of cytokines and chemokines. rhAPC alone did not have procoagulant or pro-inflammatory effects. Locally administered rhAPC has unexpected procoagulant and pro-inflammatory effects in LPS-challenged lung subsegments. These data argue against a role for intrapulmonary delivery of rhAPC as a treatment strategy for lung inflammatory disorders in humans.

Kager LM; de Boer JD; Bresser P; van der Zee JS; Zeerleder S; Meijers JC; van 't Veer C; van der Poll T

2013-07-01

309

Intrabronchial activated protein C enhances lipopolysaccharide-induced pulmonary responses.  

Science.gov (United States)

Intravenous administration of activated protein C (APC) inhibits coagulation and inflammation in the lungs of humans and animals. Investigations in rodents demonstrated that direct intrapulmonary delivery of APC also exerts anticoagulant and anti-inflammatory effects. The effect of intrabronchial administration of recombinant human (rh)APC on lipopolysaccharide (LPS)-induced haemostatic and inflammatory alterations in the bronchoalveolar space of humans was studied. Eight subjects received rhAPC via intrabronchial instillation by bronchoscope, while in a contralateral subsegment subjects received saline; all subjects were challenged bilaterally with LPS in the same lung subsegments. Four additional subjects received rhAPC (75 ?g), with saline as a control in the contralateral subsegment, while they were bilaterally "challenged" with saline. After 6 h a bronchoalveolar lavage was performed and coagulation and inflammatory parameters were measured. rhAPC enhanced LPS-induced coagulation activation in the bronchoalveolar space, when compared with the control side. In addition, rhAPC amplified LPS-induced pro-inflammatory responses, as indicated by higher concentrations of cytokines and chemokines. rhAPC alone did not have procoagulant or pro-inflammatory effects. Locally administered rhAPC has unexpected procoagulant and pro-inflammatory effects in LPS-challenged lung subsegments. These data argue against a role for intrapulmonary delivery of rhAPC as a treatment strategy for lung inflammatory disorders in humans. PMID:23060625

Kager, Liesbeth M; de Boer, J Daan; Bresser, Paul; van der Zee, Jaring S; Zeerleder, Sacha; Meijers, Joost C M; van 't Veer, Cornelis; van der Poll, Tom

2012-10-11

310

Serum lipopolysaccharide-binding protein as a marker of atherosclerosis.  

UK PubMed Central (United Kingdom)

BACKGROUND: Recently, serum lipopolysaccharide-binding protein (LBP) has been closely associated with coronary artery disease. Here, we aimed to investigate the possible relationship between serum LBP and markers of atherosclerosis. METHODS: Serum LBP and carotid intima media thickness (C-IMT) were measured in 332 subjects (101 men and 231 women) who were recruited from an ongoing multicenter project. RESULTS: Serum LBP was significantly associated with obesity [BMI, fat mass and waist circumference (r > 0.38, p < 0.0001)], HOMA (r = 0.36, p < 0.0001) and high sensitivity CRP (hsCRP) (r = 0.50, p < 0.0001). Circulating LBP was also positively associated with C-IMT (r = 0.27, p < 0.0001). Circulating LBP (? = 0.16; p = 0.001) contributed independently to C-IMT variance, after controlling for the effects of age, gender, BMI and hsCRP. Of note, circulating LBP was found to be increased in the population with carotid plaque (n = 50; 32.7 ± 12.5 vs 28.7 ± 10.7; p = 0.021). CONCLUSION: The consistent association between serum LBP and the carotid intima media thickness, a widely used atherosclerosis marker, reveals serum LBP as a putative factor related to atherosclerosis.

Serrano M; Moreno-Navarrete JM; Puig J; Moreno M; Guerra E; Ortega F; Xifra G; Ricart W; Fernández-Real JM

2013-10-01

311

Lipopolysaccharide induced inflammation in the perivascular space in lungs  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Lipopolysaccharide (LPS) contained in tobacco smoke and a variety of environmental and occupational dusts is a toxic agent causing lung inflammation characterized by migration of neutrophils and monocytes into alveoli. Although migration of inflammatory cells into alveoli of LPS-treated rats is well characterized, the dynamics of their accumulation in the perivascular space (PVS) leading to a perivascular inflammation (PVI) of pulmonary arteries is not well described. Methods Therefore, we investigated migration of neutrophils and monocytes into PVS in lungs of male Sprague-Dawley rats treated intratracheally with E. coli LPS and euthanized after 1, 6, 12, 24 and 36 hours. Control rats were treated with endotoxin-free saline. H&E stained slides were made and immunohistochemistry was performed using a monocyte marker and the chemokine Monocyte-Chemoattractant-Protein-1 (MCP-1). Computer-assisted microscopy was performed to count infiltrating cells. Results Surprisingly, the periarterial infiltration was not a constant finding in each animal although LPS-induced alveolitis was present. A clear tendency was observed that neutrophils were appearing in the PVS first within 6 hours after LPS application and were decreasing at later time points. In contrast, mononuclear cell infiltration was observed after 24 hours. In addition, MCP-1 expression was present in perivascular capillaries, arteries and the epithelium. Conclusion PVI might be a certain lung reaction pattern in the defense to infectious attacks.

Tschernig Thomas; Janardhan Kyathanahalli S; Pabst Reinhard; Singh Baljit

2008-01-01

312

Interactions of lipopolysaccharide and polymyxin studied by NMR spectroscopy.  

UK PubMed Central (United Kingdom)

In the light of occurrence of bacterial strains with multiple resistances against most antibiotics, antimicrobial peptides that interact with the outer layer of Gram-negative bacteria, such as polymyxin (PMX), have recently received increased attention. Here we present a study of the interactions of PMX-B, -E, and -M with lipopolysaccharide (LPS) from a deep rough mutant strain of Escherichia coli. A method for efficient purification of biosynthetically produced LPS using reversed-phase high-performance liquid chromatography in combination with ternary solvent mixtures was developed. LPS was incorporated into a membrane model, dodecylphosphocholine micelles, and its interaction with polymyxins was studied by heteronuclear NMR spectroscopy. Data from chemical shift mapping using isotope-labeled LPS or labeled polymyxin, as well as from isotope-filtered nuclear Overhauser effect spectroscopy experiments, reveal the mode of interaction of LPS with polymyxins. Using molecular dynamics calculations the complex of LPS with PMX-B in the presence of dodecylphosphocholine micelles was modeled using restraints derived from chemical shift mapping data and intermolecular nuclear Overhauser effects. In the modeled complex the macrocycle of PMX is centered around the phosphate group at GlcN-B, and additional contacts from polar side chains are formed to GlcN-A and Kdo-C, whereas hydrophobic side chains penetrate the acyl-chain region.

Mares J; Kumaran S; Gobbo M; Zerbe O

2009-04-01

313

Lipopolysaccharide-induced dental pulp cell apoptosis and the expression of Bax and Bcl-2 in vitro  

Directory of Open Access Journals (Sweden)

Full Text Available Lipopolysaccharide exerts many effects on many cell lines, including cytokine secretion, and cell apoptosis and necrosis. We investigated the in vitro effects of lipopolysaccharide on apoptosis of cultured human dental pulp cells and the expression of Bcl-2 and Bax. Dental pulp cells showed morphologies typical of apoptosis after exposure to lipopolysaccharide. Flow cytometry showed that the rate of apoptosis of human dental pulp cells increased with increasing lipopolysaccharide concentration. Compared with controls, lipopolysaccharide promoted pulp cell apoptosis (P < 0.05) from 0.1 to 100 ?g/mL but not at 0.01 ?g/mL. Cell apoptosis was statistically higher after exposure to lipopolysaccharide for 3 days compared with 1 day, but no difference was observed between 3 and 5 days. Immunohistochemistry showed that expression of Bax and Bcl-2 was enhanced by lipopolysaccharide at high concentrations, but no evident expression was observed at low concentrations (0.01 and 0.1 ?g/mL) or in the control groups. In conclusion, lipopolysaccharide induced dental pulp cell apoptosis in a dose-dependent manner, but apoptosis did not increase with treatment duration. The expression of the apoptosis regulatory proteins Bax and Bcl-2 was also up-regulated in pulp cells after exposure to a high concentration of lipopolysaccharide.

H. Yang; Y.T. Zhu; R. Cheng; M.Y. Shao; Z.S. Fu; L. Cheng; F.M. Wang; T. Hu

2010-01-01

314

Oil field and freshwater isolates of Shewanella putrefaciens have lipopolysaccharide polyacrylamide gel profiles characteristic of marine bacteria  

Energy Technology Data Exchange (ETDEWEB)

The lipopolysaccharide structure of oil field and freshwater isolates of bacteria that reduce ferric iron, recently classified as strains of Shewanella putrefaciens, was analyzed using polyacrylamide gel electrophoresis and a lipopolysaccharide-specific silver-staining procedure. The results demonstrate that all the oil field and freshwater isolates examined exhibited the more hydrophobic R-type lipopolysaccharide, which has been found to be characteristic of Gram-negative marine bacteria. This hydrophobic lipopolysaccharide would confer an advantage on bacteria involved in hydrocarbon degradation by assisting their association with the surface of oil droplets. 15 refs., 1 fig.

Pickard, C.; Foght, J.M.; Pickard, M.A.; Westlake, D.W.S. (Alberta Univ., Edmonton, AB (Canada))

1993-01-01

315

Methylprednisolone Stiffens Aortas in Lipopolysaccharide-Induced Chronic Inflammation in Rats  

Science.gov (United States)

Introduction Glucocorticoids are commonly used as therapeutic agents in many acute and chronic inflammatory and auto-immune diseases. The current study investigated the effects of methylprednisolone (a synthetic glucocorticoid) on aortic distensibility and vascular resistance in lipopolysaccharide-induced chronic inflammation in male Wistar rats. Methods Chronic inflammation was induced by implanting a subcutaneous slow-release ALZET osmotic pump (1 mg kg?1 day?1 lipopolysaccharide) for either 2 or 4 weeks. Arterial wave transit time (?) was derived to describe the elastic properties of aortas using the impulse response function of the filtered aortic input impedance spectra. Results Long-term lipopolysaccharide challenge enhanced the expression of advanced glycation end products (AGEs) in the aortas. Lipopolysaccharide also upregulated the inducible form of nitric oxide synthase to produce high levels of nitric oxide (NO), which resulted in vasodilation, as evidenced by the fall in total peripheral resistance (Rp). However, lipopolysaccharide challenge did not influence the elastic properties of aortas, as shown by the unaltered ?. The NO-mediated vascular relaxation may counterbalance the AGEs-induced arterial stiffening so that the aortic distensibility remained unaltered. Treating lipopolysaccharide-challenged rats with methylprednisolone prevented peripheral vasodilation because of its ability to increase Rp. However, methylprednisolone produced an increase in aorta stiffness, as manifested by the significant decline in ?. The diminished aortic distensibility by methylprednisolone paralleled a significant reduction in NO plasma levels, in the absence of any significant changes in AGEs content. Conclusion Methylprednisolone stiffens aortas and elastic arteries in lipopolysaccharide-induced chronic inflammation in rats, for NO activity may be dominant as a counteraction of AGEs.

Ko, Ya-Hui; Tsai, Ming-Shian; Lee, Po-Huang; Liang, Jin-Tung; Chang, Kuo-Chu

2013-01-01

316

Glucocorticoid-dependent and -independent mechanisms involved in lipopolysaccharide tolerance.  

UK PubMed Central (United Kingdom)

Injection of bacterial lipopolysaccharide (LPS) into animals results in a transient increase in serum tumor necrosis factor (TNF). Maximal increases in TNF were detected by 1 h and 3-4 h serum TNF was no longer apparent. These animals were LPS tolerant and a repetitive LPS stimulus did not result in an additional peak in TNF. Regulation of TNF expression in LPS-tolerant animals was at the transcriptional level as TNF mRNA was not apparent in spleen or peritoneal macrophages following a second LPS stimulus. Adrenalectomized (adrex) mice, in contrast, did not become LPS tolerant and sera from these animals demonstrated an additional peak in TNF 1 h following a second LPS stimulus. Concomitant with the secondary rise in serum TNF in adrex mice was an increase in splenic TNF mRNA. The ability of adrex mice to become LPS tolerant was restored by exogenous glucocorticoids. LPS tolerance was also investigated in the galactosamine LPS model which like the adrex model is characterized by a thousandfold increase in the sensitivity of these animals to the lethal effects of LPS. Consistent with the absence of LPS tolerance in adrex mice, galactosamine-sensitized mice were also responsive to a second LPS stimulus and did not become LPS tolerant. While LPS-treated adrex mice had no significant increases in serum corticosterone, corticosterone levels in LPS-treated galactosamine-sensitized mice were comparable to LPS-stimulated normals suggesting that LPS tolerance involves both glucocorticoid-dependent and -independent components. Finally, prophylactic administration of a monoclonal antibody against murine TNF protected normal and galactosamine-sensitized mice from a lethal dose of LPS and yet had no protective effect in adrex animals.

Evans GF; Zuckerman SH

1991-09-01

317

Lipopolysaccharide induces a fibrotic-like phenotype in endothelial cells.  

UK PubMed Central (United Kingdom)

Endothelial dysfunction is crucial in endotoxaemia-derived sepsis syndrome pathogenesis. It is well accepted that lipopolysaccharide (LPS) induces endothelial dysfunction through immune system activation. However, LPS can also directly generate actions in endothelial cells (ECs) in the absence of participation by immune cells. Although interactions between LPS and ECs evoke endothelial death, a significant portion of ECs are resistant to LPS challenge. However, the mechanism that confers endothelial resistance to LPS is not known. LPS-resistant ECs exhibit a fibroblast-like morphology, suggesting that these ECs enter a fibrotic programme in response to LPS. Thus, our aim was to investigate whether LPS is able to induce endothelial fibrosis in the absence of immune cells and explore the underlying mechanism. Using primary cultures of ECs and culturing intact blood vessels, we demonstrated that LPS is a crucial factor to induce endothelial fibrosis. We demonstrated that LPS was able and sufficient to promote endothelial fibrosis, in the absence of immune cells through an activin receptor-like kinase 5 (ALK5) activity-dependent mechanism. LPS-challenged ECs showed an up-regulation of both fibroblast-specific protein expression and extracellular matrix proteins secretion, as well as a down-regulation of endothelial markers. These results demonstrate that LPS is a crucial factor in inducing endothelial fibrosis in the absence of immune cells through an ALK5-dependent mechanism. It is noteworthy that LPS-induced endothelial fibrosis perpetuates endothelial dysfunction as a maladaptive process rather than a survival mechanism for protection against LPS. These findings are useful in improving current treatment against endotoxaemia-derived sepsis syndrome and other inflammatory diseases.

Echeverría C; Montorfano I; Sarmiento D; Becerra A; Nuñez-Villena F; Figueroa XF; Cabello-Verrugio C; Elorza AA; Riedel C; Simon F

2013-06-01

318

Lipopolysaccharide enhances the cytotoxicity of 2-chloroethyl ethyl sulfide  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background The bacterial endotoxin, lipopolysaccharide (LPS), is a well-characterized inflammatory factor found in the cell wall of Gram-negative bacteria. In this investigation, we studied the cytotoxic interaction between 2-chloroethyl ethyl sulfide (CEES or ClCH2CH2SCH2CH3) and LPS using murine RAW264.7 macrophages. CEES is a sulfur vesicating agent and is an analog of 2,2'-dichlorodiethyl sulfide (sulfur mustard). LPS is a ubiquitous natural agent found in the environment. The ability of LPS and other inflammatory agents (such as TNF-alpha and IL-1beta) to modulate the toxicity of CEES is likely to be an important factor in the design of effective treatments. Results RAW 264.7 macrophages stimulated with LPS were found to be more susceptible to the cytotoxic effect of CEES than unstimulated macrophages. Very low levels of LPS (20 ng/ml) dramatically enhanced the toxicity of CEES at concentrations greater than 400 ?M. The cytotoxic interaction between LPS and CEES reached a maximum 12 hours after exposure. In addition, we found that tumor necrosis factor-alpha (TNF-alpha) and interleukin-1-beta (IL-1-beta) as well as phorbol myristate acetate (PMA) also enhanced the cytotoxic effects of CEES but to a lesser extent than LPS. Conclusion Our in vitro results suggest the possibility that LPS and inflammatory cytokines could enhance the toxicity of sulfur mustard. Since LPS is a ubiquitous agent in the natural environment, its presence is likely to be an important variable influencing the cytotoxicity of sulfur mustard toxicity. We have initiated further experiments to determine the molecular mechanism whereby the inflammatory process influences sulfur mustard cytotoxicity.

Stone William L; Qui Min; Smith Milton

2003-01-01

319

Shikonin attenuates lipopolysaccharide-induced acute lung injury in mice.  

UK PubMed Central (United Kingdom)

BACKGROUND: Shikonin, a natural naphthoquinone pigment extracted from the root of Lithospermum erythrorhizon, has shown a variety of pharmacologic properties including anti-inflammatory effect. In the present study, we analyzed the role of shikonin in acute lung injury induced by lipopolysaccharide (LPS) in mice. MATERIALS AND METHODS: Sixty male BALB/C mice were randomly allocated into six groups (n = 10, each): control group, shikonin group (50 mg/kg), LPS group, and three different doses (12.5, 25, and 50 mg/kg) for shikonin-treated groups. Shikonin or vehicle was given with an intragastric administration 1 h before an intratracheal instillation of LPS (5 mg/kg). The severity of pulmonary injury was evaluated 6 h after LPS challenge. RESULTS: Shikonin pretreatment significantly attenuated LPS-induced pulmonary histopathologic changes, alveolar hemorrhage, and neutrophil infiltration. The lung wet-to-dry weight ratios, as the index of pulmonary edema, were markedly decreased by shikonin pretreatment. Moreover, shikonin decreased the productions of the proinflammatory cytokines including tumor necrosis factor alpha and interleukin 1? and the concentration of total proteins in the bronchoalveolar lavage fluid. Shikonin pretreatment also reduced the concentrations of myeloperoxidase and nitric oxide in lung tissues. In addition, shikonin pretreatment significantly suppressed LPS-induced activation of cyclooxygenase 2 and inducible nitric oxide synthase and the nuclear factor ?B DNA-binding activity in lung tissues. CONCLUSIONS: This study indicates that shikonin may have a protective effect against LPS-induced acute lung injury, and the potential mechanism of this action may attribute partly to the inhibition of inducible nitric oxide synthase and cyclooxygenase 2 expression by downregulating nuclear factor ?B activation.

Bai GZ; Yu HT; Ni YF; Li XF; Zhang ZP; Su K; Lei J; Liu BY; Ke CK; Zhong DX; Wang YJ; Zhao JB

2013-06-01

320

HORMONAL SYNCHRONIZATION OF LIPOPOLYSACCHARIDE-INDUCED HYPOTHERMIC RESPONSE IN RATS.  

UK PubMed Central (United Kingdom)

Background: Recent experimental evidence suggests that lipopolysaccharide (LPS)- induced hypothermia is an adaptive thermoregulatory strategy against immunological challenge in rats. We hypothesized that the hormones which are predominantly responsible for energy homeostasis may have efferent signaling roles for development of the hypothermia. Aim: The aim of the study was to evaluate the changes of hypothalamic-pituitary-thyroid (HPT) and hypothalamic-pituitary-adrenal (HPA) axis hormones, leptin and erythropoietin at various phases of LPS-induced hypothermia such as the initial phase, nadir and the end of the response in blood sampled rats. Material and Methods: Body temperature of adult male albino Wistar rats was recorded by biotelemetry. E.coli O111:B4 LPS (250 Ag/kg, ip) was injected alone or with SC-560, a cyclooxygenase-1 selective inhibitor (1 mg/kg, sc). Results: Serum FT4 levels elevated at the initial phase, but FT3 levels decreased at nadir and remained low at the end of the response. Meanwhile, no change was observed in TSH levels. Serum adrenocorticotropic hormone (ACTH) levels reduced at the initial phase and serum corticosterone levels decreased at nadir without any change in serum corticotropin-releasing hormone (CRH) levels throughout the hypothermia. Serum leptin levels increased only at the end of the response. No change was observed in the levels of serum erythropoietin. SC-560 treatment abolished both LPS-induced hypothermia and respective hormonal changes. Conclusion: Data suggest that HPT axis hormones may contribute for development of LPS-induced hypothermia in rats. Data also support the view that leptin may have a role for the recovery of hypothermic response.

Polat H; Mamuk S; Akarsu ES

2013-04-01

 
 
 
 
321

[Lipopolysaccharide induced expression of peroxiredoxin 1 in airway epithelial cell].  

UK PubMed Central (United Kingdom)

OBJECTIVE: To investigate the effect of lipopolysaccharide ( LPS) on expression of peroxiredoxin 1(prdx1) in airway epithelial cells. METHODS: The airway epithelium cell line BEAS-2B was cultivated, and the cells were stimulated with 0, 1, and 10 mg/L of LPS for 12 hours and 24 hours, and then were harvested for prdx1 expression detection. The mRNA expression of prdx1 was detected by reverse transcription-polymerase chain reaction (RT-PCR).The airway epithelium cells were stimulated with 0, 0.1 , 0.5, 1 , 5, and 10 mg/L of LPS for 12 hours, and were collected for determination of prdx 1 protein expression by Western blotting. RESULTS: RT-PCR results showed that the prdx1 mRNA expression was significantly increased within 12 hours of stimulation with elevation of the dosage of LPS.The prdx1 mRNA expression at 12 hours of stimulation by 10 mg/L LPS was significantly higher than that in control group(2.014 ± 0.197 vs. 0.644 ± 0.178, P<0.05). However, with prolongation of LPS stimulation time, the prdx1 mRNA expression at 24 hours was slightly declined. Western blotting results showed that the prdx1 protein expression was gradually increased with elevation of dosage of LPS. The prdx1 protein expression at 12 hours of stimulation with 5 mg/L LPS was significantly higher than that in control group ( 1.069 ± 0.175 vs. 0.328 ± 0.010, P<0.05), and the expression remained at high level at 12 hours of stimulation with 10 mg/L LPS (0.984 ± 0.220 ). CONCLUSION: 10 mg/Lof LPS can induce the mRNA and protein expression of prdx1 in BEAS-2B cell after 12 hours of stimulation.

Liu Yi; Mao P; Liu XQ; He WQ; Mo HY; Li YM

2013-03-01

322

Lipopolysaccharide induces a fibrotic-like phenotype in endothelial cells.  

Science.gov (United States)

Endothelial dysfunction is crucial in endotoxaemia-derived sepsis syndrome pathogenesis. It is well accepted that lipopolysaccharide (LPS) induces endothelial dysfunction through immune system activation. However, LPS can also directly generate actions in endothelial cells (ECs) in the absence of participation by immune cells. Although interactions between LPS and ECs evoke endothelial death, a significant portion of ECs are resistant to LPS challenge. However, the mechanism that confers endothelial resistance to LPS is not known. LPS-resistant ECs exhibit a fibroblast-like morphology, suggesting that these ECs enter a fibrotic programme in response to LPS. Thus, our aim was to investigate whether LPS is able to induce endothelial fibrosis in the absence of immune cells and explore the underlying mechanism. Using primary cultures of ECs and culturing intact blood vessels, we demonstrated that LPS is a crucial factor to induce endothelial fibrosis. We demonstrated that LPS was able and sufficient to promote endothelial fibrosis, in the absence of immune cells through an activin receptor-like kinase 5 (ALK5) activity-dependent mechanism. LPS-challenged ECs showed an up-regulation of both fibroblast-specific protein expression and extracellular matrix proteins secretion, as well as a down-regulation of endothelial markers. These results demonstrate that LPS is a crucial factor in inducing endothelial fibrosis in the absence of immune cells through an ALK5-dependent mechanism. It is noteworthy that LPS-induced endothelial fibrosis perpetuates endothelial dysfunction as a maladaptive process rather than a survival mechanism for protection against LPS. These findings are useful in improving current treatment against endotoxaemia-derived sepsis syndrome and other inflammatory diseases. PMID:23635013

Echeverría, César; Montorfano, Ignacio; Sarmiento, Daniela; Becerra, Alvaro; Nuñez-Villena, Felipe; Figueroa, Xavier F; Cabello-Verrugio, Claudio; Elorza, Alvaro A; Riedel, Claudia; Simon, Felipe

2013-05-02

323

GLYCOLIPID FRACTION FROM CYANOBACTERIA FOR TREATMENT OF DISEASES OF THE ORAL CAVITY  

UK PubMed Central (United Kingdom)

The present invention relates to the preparation and use of a glycolipid fraction from Oscillatoria Planktothrix sp., for the treatment and/or prevention of bacterial gum diseases primarily caused by: Actinobacillum actinomycetemconcomitans, Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola and even more preferably by Porphyromonas gingivalis. Said gum diseases, in particular gingivitis and periodontitis (pyorrhea), are primarily caused by a pro-inflammatory response to components of P. gingivalis, leading to destruction of periodontal tissue, and are often accompanied by osteoclastogenesis (increased number of osteoclasts responsible for destruction of bone tissue), and by chronic infection.

MOLTENI MONICA

324

Quantitative determination of ion distributions in bacterial lipopolysaccharide membranes by grazing-incidence X-ray fluorescence  

Digital Repository Infrastructure Vision for European Research (DRIVER)

A model of the outer membrane of Gram-negative bacteria was created by the deposition of a monolayer of purified rough mutant lipopolysaccharides at an air/water interface. The density profiles of monovalent (K+) and divalent (Ca2+) cations normal to the lipopolysaccharides (LPS) monolayers were inv...

Schneck, Emanuel; Schubert, Thomas; Konovalov, Oleg V.; Quinn, Bonnie E.; Gutsmann, Thomas; Brandenburg, Klaus

325

Variation in the structure and bacteriophage-inactivating capacity of Salmonella anatum lipopolysaccharide as a function of growth temperature.  

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Growth temperature affects both the structure and the phage-inactivating capacity of Salmonella anatum A1 lipopolysaccharide. Whereas S. anatum cells normally synthesize smooth lipopolysaccharide when grown at physiological temperature (37 degrees C), a partial smooth-rough transition occurs when ce...

McConnell, M; Wright, A

326

Baclofen influences lipopolysaccharide-mediated interleukin-6 release from murine pituicytes  

DEFF Research Database (Denmark)

Pituicytes, the glial cells of the neurohypophysis, secrete interleukin-6 upon stimulation with various inflammatory mediators, i.e. lipopolysaccharide. Previous studies have identified several receptors on pituicytes. This study investigates the effect of GABA(B) receptor activation on interleukin-6 release from pituicytes. Cultured murine pituicytes were stimulated for 24 h with lipopolysaccharide (0.5 ng/ml) to give a significant interleukin-6 release compared to control. The interleukin-6 release was significantly potentiated by the GABA(B) receptor agonist (R)-4-amino-3-(4-chlorophenyl) butanoic acid (R-baclofen; 10, 100 or 500 microM). However, R-baclofen itself (10, 100 or 500 microM) did not stimulate the interleukin-6 secretion. Furthermore, the potent GABA(B) receptor antagonists 3-[[(3,4-Dichlorophenyl)methyl]amino]propyl]diethoxymethyl) phosphinic acid (CGP52432; 30 or 300 microM) and (RS)-3-Amino-2-(4-chlorophenyl)-2-hydroxypropyl-sulphonic acid (2-OH-saclofen; 10 or 100 microM) did not remove the effect of R-baclofen (100 microM). Gamma-amino butyric acid (GABA; 30 or 300 microM) did not alter the lipopolysaccharide-mediated interleukin-6 response. After 30 min, intracellular cyclic AMP (cAMP) was higher in cells stimulated with lipopolysaccharide compared to control, and R-baclofen significantly inhibited this increase in cAMP. Nevertheless, neither lipopolysaccharide nor R-baclofen had any effect on intracellular cAMP after 24 h of stimulation. The results suggest that the effect of R-baclofen on lipopolysaccharide-stimulated interleukin-6 secretion is independent of GABA(B) receptors.

Kjeldsen, Tine H; Hansen, Erik W

2002-01-01

327

Dry-heat destruction of lipopolysaccharide: mathematical approach to process evaluation.  

UK PubMed Central (United Kingdom)

A mathematical model was developed to estimate the number of logarithmic cycles (LDec) of lipopolysaccharide concentration destroyed by a dry-heat sterilization process. The LDec values calculated from the mathematical model agreed well with those obtained from the destruction of lipopolysaccharide by a dry-heat treatment. A discussion of how the mathematical model may be used to evaluate a dry-heat sterilization cycle is presented. This mathematical model and the dry-heat destruction curves indicated existence of a maximum LDec value at each temperature. The implications of this finding are discussed.

Tsuji K; Lewis AR

1978-11-01

328

Lipid lateral organization on giant unilamellar vesicles containing lipopolysaccharides  

DEFF Research Database (Denmark)

We developed a new (to our knowledge) protocol to generate giant unilamellar vesicles (GUVs) composed of mixtures of single lipopolysaccharide (LPS) species and Escherichia coli polar lipid extracts. Four different LPSs that differed in the size of the polar headgroup (i.e., LPS smooth > LPS-Ra > LPS-Rc > LPS-Rd) were selected to generate GUVs composed of different LPS/E. coli polar lipid mixtures. Our procedure consists of two main steps: 1), generation and purification of oligolamellar liposomes containing LPSs; and 2), electroformation of GUVs using the LPS-containing oligolamellar vesicles at physiological salt and pH conditions. Analysis of LPS incorporation into the membrane models (both oligolamellar vesicles and GUVs) shows that the final concentration of LPS is lower than that expected from the initial E. coli lipids/LPS mixture. In particular, our protocol allows incorporation of no more than 15 mol % for LPS-smooth and LPS-Ra, and up to 25 mol % for LPS-Rc and LPS-Rd (with respect to total lipids).We used the GUVs to evaluate the impact of different LPS species on the lateral structure of the host membrane (i.e., E. coli polar lipid extract). Rhodamine-DPPE-labeled GUVs show the presence of elongated micrometer-sized lipid domains for GUVs containing either LPS-Rc or LPS-Rd above 10 mol %. Laurdan GP images confirm this finding and show that this particular lateral scenario corresponds to the coexistence of fluid disordered and gel (LPS-enriched)-like micron-sized domains, in similarity to what is observed when LPS is replaced with lipid A. For LPSs containing the more bulky polar headgroup (i.e., LPS-smooth and LPS-Ra), an absence of micrometer-sized domains is observed for all LPS concentrations explored in the GUVs (up to ?15 mol %). However, fluorescence correlation spectroscopy (using fluorescently labeled LPS) and Laurdan GP experiments in these microscopically homogeneous membranes suggests the presence of LPS clusters with dimensions below our microscope's resolution (?380 nm radial). Our results indicate that LPSs can cluster into gel-like domains in these bacterial model membranes, and that the size of these domains depends on the chemical structure and concentration of the LPSs.

Kubiak, Jakub; Brewer, Jonathan R.

2011-01-01

329

Comparing immune activation (lipopolysaccharide) and toxin (lithium chloride)-induced gustatory conditioning: lipopolysaccharide produces conditioned taste avoidance but not aversion.  

UK PubMed Central (United Kingdom)

Feeding and drinking typically involve both appetitive and consummatory behaviors. Appetitive behaviors include those behaviors produced by an animal prior to the actual consumption, such as approach movements, whereas consummatory behaviors (such as licking and chewing) are involved in the actual consumption of food. The present research compared the gustatory conditioning effects of bacterial lipopolysaccharide (LPS) and lithium chloride (LiCl) in two different paradigms, conditioned taste avoidance and conditioned taste aversion which differentially affect the appetitive and consummatory components of feeding. Male rats were implanted with intraoral cannulae and habituated to a water deprivation schedule and afterwards received two conditioning days (Days 1 and 4). Each conditioning day consisted of 1 h access to a novel sucrose solution (0.3 M) immediately followed by a systemic injection of LPS (200 microg/kg), LiCl (0.15 M, 3 meq) or NaCl vehicle. Conditioned taste aversion was assessed using the taste reactivity test on Day 7, where orofacial and somatic responses were videotaped and analyzed during 3 brief (1 min) exposures to the sucrose solution. Conditioned taste avoidance was assessed on Days 8 and 9 using a two-bottle preference test (sucrose versus water). Animals conditioned with LiCl displayed typical aversive-like responses in the taste reactivity paradigm evidenced by significant reductions in positive ingestive responses (P<0.05) and an increase in active aversive responses (P<0.05) relative to controls. Furthermore, LiCl treatment resulted in conditioned avoidance of sucrose in the two-bottle preference test characterized by a decreased sucrose preference (P<0.05). Conditioning with LPS produced a reduced sucrose preference (P<0.05) relative to controls, comparable to the avoidance seen in LiCl-treated rats. In contrast, conditioning with LPS resulted in similar positive ingestive responses to intraorally infused sucrose as seen in controls. The present results demonstrate that LPS treatment produces conditioned avoidance but not aversion and suggest that LPS can selectively condition the appetitive aspects of feeding whereas the consummatory behaviors remain unaffected.

Cross-Mellor SK; Kavaliers M; Ossenkopp KP

2004-01-01

330

Comparing immune activation (lipopolysaccharide) and toxin (lithium chloride)-induced gustatory conditioning: lipopolysaccharide produces conditioned taste avoidance but not aversion.  

Science.gov (United States)

Feeding and drinking typically involve both appetitive and consummatory behaviors. Appetitive behaviors include those behaviors produced by an animal prior to the actual consumption, such as approach movements, whereas consummatory behaviors (such as licking and chewing) are involved in the actual consumption of food. The present research compared the gustatory conditioning effects of bacterial lipopolysaccharide (LPS) and lithium chloride (LiCl) in two different paradigms, conditioned taste avoidance and conditioned taste aversion which differentially affect the appetitive and consummatory components of feeding. Male rats were implanted with intraoral cannulae and habituated to a water deprivation schedule and afterwards received two conditioning days (Days 1 and 4). Each conditioning day consisted of 1 h access to a novel sucrose solution (0.3 M) immediately followed by a systemic injection of LPS (200 microg/kg), LiCl (0.15 M, 3 meq) or NaCl vehicle. Conditioned taste aversion was assessed using the taste reactivity test on Day 7, where orofacial and somatic responses were videotaped and analyzed during 3 brief (1 min) exposures to the sucrose solution. Conditioned taste avoidance was assessed on Days 8 and 9 using a two-bottle preference test (sucrose versus water). Animals conditioned with LiCl displayed typical aversive-like responses in the taste reactivity paradigm evidenced by significant reductions in positive ingestive responses (P<0.05) and an increase in active aversive responses (P<0.05) relative to controls. Furthermore, LiCl treatment resulted in conditioned avoidance of sucrose in the two-bottle preference test characterized by a decreased sucrose preference (P<0.05). Conditioning with LPS produced a reduced sucrose preference (P<0.05) relative to controls, comparable to the avoidance seen in LiCl-treated rats. In contrast, conditioning with LPS resulted in similar positive ingestive responses to intraorally infused sucrose as seen in controls. The present results demonstrate that LPS treatment produces conditioned avoidance but not aversion and suggest that LPS can selectively condition the appetitive aspects of feeding whereas the consummatory behaviors remain unaffected. PMID:14684243

Cross-Mellor, Shelley K; Kavaliers, Martin; Ossenkopp, Klaus-Peter

2004-01-01

331

Calcium ions induce collapse of charged O-side chains of lipopolysaccharides from Pseudomonas aeruginosa  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Lipopolysaccharide (LPS) monolayers deposited on planar, hydrophobic substrates were used as a defined model of outer membranes of Pseudomonas aeruginosa strain dps 89. To investigate the influence of ions on the (out-of-plane) monolayer structure, we measured specular X-ray reflectivity at high ene...

Schneck, Emanuel; Papp-Szabo, Erzsebet; Quinn, Bonnie E.; Konovalov, Oleg V.; Beveridge, Terry J.; Pink, David A.

332

Experimental induction of recurrent airway obstruction with inhaled fungal spores, lipopolysaccharide, and silica microspheres in horses.  

UK PubMed Central (United Kingdom)

OBJECTIVE: To evaluate experimental induction of recurrent airway obstruction (RAO) with inhaled fungal spores, lipopolysaccharide, and silica microspheres in horses. ANIMALS: 7 horses with and 3 horses without a history of RAO. PROCEDURES: RAO-susceptible horses ranged in age from 17 to approximately 30 years, and control horses ranged in age from 7 to approximately 15 years. Pure mold cultures were derived from repeated culture of hay and identified via gene amplification and sequencing. Pulmonary function testing and bronchoalveolar lavage were performed before and after nebulization with a suspension of spores derived from 3 fungi, lipopolysaccharide, and 1-microm silica microspheres in all horses. This was followed by a 4-month washout period and a further pulmonary function test followed by saline (0.9% NaCl) solution challenge and bronchoalveolar lavage. RESULTS: Lichtheimia corymbifera, Aspergillus fumigatus, and Eurotium amstelodami were consistently identified in cultures of moldy hay. Nebulization with fungal spores, lipopolysaccharide, and microspheres induced significant increases in pleural pressure in RAO-susceptible but not control horses. Airway neutrophilia developed in both groups of horses with exposure to challenge material but more severely in RAO-susceptible horses. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that inhalation of fungal spores in combination with lipopolysaccharide and silica microspheres can induce disease exacerbation in susceptible horses and may thus be a useful model for future standardized studies of RAO in horses.

Beeler-Marfisi J; Clark ME; Wen X; Sears W; Huber L; Ackerley C; Viel L; Bienzle D

2010-06-01

333

Regulation of hepcidin and ferroportin expression by lipopolysaccharide in splenic macrophages.  

UK PubMed Central (United Kingdom)

Acute and chronic inflammatory states are associated with many changes in intracellular iron metabolism including sequestration of iron in the mononuclear-phagocyte system (MPS) and a decline in serum iron. Previous work in rodent models of acute inflammation has demonstrated inflammation-induced downregulation of intestinal and MPS iron exporter, ferroportin 1, mRNA and protein. In addition, these models have also demonstrated hepatic induction of mRNA of the small 25 amino acid peptide hepcidin. Hepcidin has been hypothesized to be the mediator of iron- and inflammation-induced changes in iron metabolism. The molecular details of the connection between iron metabolism, hepcidin and inflammation have become clearer with the recent finding of hepcidin-induced internalization and degradation of FPN1. The work presented here demonstrates that the lipopolysaccharide-induced splenic macrophage FPN1 mRNA downregulation is not dependent upon the action of a single cytokine such as IL-6, IL-1 or TNF-alpha because mice deficient in these pathways downregulate FPN1 normally. Furthermore, hepcidin is also synthesized in the spleen of normal mice and induced by lipopolysaccharide. Additionally, in vitro, splenic adherent cells produce hepcidin in response to lipopolysaccharide in an IL-6-dependent manner. There appear to be both probable transcriptional and post-transcriptional control of FPN1 expression by lipopolysaccharide-induced inflammation. The former effect is on mRNA expression and is independent of hepcidin, whereas the latter is IL-6- and hepcidin-dependent.

Liu XB; Nguyen NB; Marquess KD; Yang F; Haile DJ

2005-07-01

334

[Estimation of mean molecular weight of Enterobacteriaceae lipopolysaccharides using gas chromatography for determination of fatty acids].  

Science.gov (United States)

In the paper, we propose a method for estimation of the mean molecular weight of lipopolysaccharide, which is important for accuracy of endotoxin activity investigation. In our study, it was assumed that lipid A portion in Enterobacterial lipopolysaccharide is substituted by four 3-hydroxytetradecanoic acid residues. Lipopolysaccharides of S, Ra, Rc and Re chemotypes being laboratory preparations as well as purchased from Sigma were investigated. Fatty acids were determined by of gas chromatography as methyl esters according to the procedure described by Wollenweber and Rietschel. Mean molecular weight was calculated by the formula: MMW = [formula: see text]. A high agreement between the estimated and the theoretical molecular weight values was demonstrated in the case of Salmonella minnesota R595 (Re) LPS preparation. As expected, LPS heterogeneity increase together with enlargement of polysaccharide chain length which is visible in electrophoregrams also. Except for LPS mean molecular weight estimation, the method allows its detection in various preparations and samples, distinguishing of R and S LPS forms as well as the determination of mean length of O-specific chain in lipopolysaccharides which structures are known. PMID:10222736

Zió?kowski, A; Swierzko, A S; Cedzy?ski, M

1998-01-01

335

Heat shock protein 90 mediates macrophage activation by Taxol and bacterial lipopolysaccharide  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Taxol, a plant-derived antitumor agent, stabilizes microtubules. Taxol also elicits cell signals in a manner indistinguishable from bacterial lipopolysaccharide (LPS). LPS-like actions of Taxol are controlled by the lps gene and are independent of binding to the known Taxol target, ?-tubulin. Using ...

Byrd, Cynthia A.; Bornmann, William; Erdjument-Bromage, Hediye; Tempst, Paul; Pavletich, Nikola; Rosen, Neal; Nathan, Carl F.

336

EFFECTS OF DIESEL EXHAUST PARTICLES ON HUMAN ALVEOLAR MACROPHAGE RESPONSIVENESS TO LIPOPOLYSACCHARIDE  

Science.gov (United States)

Effects of diesel exhaust particles on human alveolar macrophage responsiveness to lipopolysaccharide S. Mundandhara1 , S. Becker2 and M. Madden2, 1UNC Center for Environmental Medicine, Asthma, and Lung Biology, 2US EPA, NHEERL, HSD, Chapel Hill, NC, US Epidemiological...

337

Nitric oxide mediates prostaglandins' deleterious effect on lipopolysaccharide-triggered murine fetal resorption  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Genital tract bacterial infections could induce abortion and are some of the most common complications of pregnancy; however, the mechanisms remain unclear. We investigated the role of prostaglandins (PGs) in the mechanism of bacterial lipopolysaccharide (LPS)-induced pregnancy loss in a mouse model...

Aisemberg, J.; Vercelli, C.; Billi, S.; Ribeiro, M. L.; Ogando, D.; Meiss, R.; McCann, S. M.; Rettori, V.; Franchi, A. M.

338

Uncoupling protein 2 plays an important role in nitric oxide production of lipopolysaccharide-stimulated macrophages  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The expression of uncoupling protein 2 (UCP2) was reduced in macrophages after stimulation with lipopolysaccharide (LPS). The physiological consequence and the regulatory mechanisms of the UCP2 down-regulation by LPS were investigated in a macrophage cell line, RAW264 cells. UCP2 overexpression in R...

Kizaki, Takako; Suzuki, Kenji; Hitomi, Yoshiaki; Taniguchi, Naoyuki; Saitoh, Daizoh; Watanabe, Kenji; Onoé, Kazunori

339

Effect of NO donor sodium nitroprusside on lipopolysaccharide induced acute lung injury in rats  

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Nitric oxide (NO) donor-sodium nitroprusside (SNP) mitigates acute lung injury (ALI), but the mechanism of this protection is incompletely known. We investigated the effect of SNP on lipopolysaccharide (LPS)-induced ALI in rats. Forty-eight male Wistar rats were randomly assigned into six groups: th...

Xia, Zy; Wang, Xy; Chen, X; Xia, Z

340

Sinigrin suppresses nitric oxide production in rats administered intraperitoneally with lipopolysaccharide  

UK PubMed Central (United Kingdom)

Sinigrin, a glucosinolate contained in cruciferous vegetables, is converted to allyl isothiocyanate by plant myrosinase or microbial myrosinase in gut microflora. The intake of sinigrin significantly decreases the urinary nitrate+nitrite level (a nitric oxide [NO] production marker) in rats administered intraperitoneally with lipopolysaccharide (LPS). This result demonstrates that sinigrin suppresses NO production and therefore likely has an antioxidative effect in vivo.

Ippoushi Katsunari; Takeuchi Atsuko; Azuma Keiko

2010-06-01

 
 
 
 
341

[Role of lipopolysaccharide in the action of complement on gram negative bacteria  

UK PubMed Central (United Kingdom)

In this review a short description of the methods for the activation of the complement system and data on the role of different structures of lipopolysaccharide of gram negative bacteria in this process are presented. Variants of complement-induced bacteriolysis are considered. Special attention is given to cholera infection and the role of Vibrio cholerae O139 polysaccharide in interaction with the complement.

Lobanov VV

2004-09-01

342

Ganglioside mimicry of Campylobacter jejuni lipopolysaccharides determines antiganglioside specificity in rabbits  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The core oligosaccharides of Campylobacter jejuni lipopolysaccharides (LPS) display molecular mimicry with gangliosides. Cross-reactive anti-LPS-antiganglioside antibodies have been implicated to show a crucial role in the pathogenesis of the Guillain-Barre and Miller Fisher syndro...

Ang, C.W.; Noordzij, P.G.; Klerk, M.A. de; Endtz, H.P.; Doorn, P.A. van; Laman, J.D.

343

Absence of a characteristic cell wall lipopolysaccharide in the phototrophic bacterium Chloroflexus aurantiacus.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Two strains of the gliding phototrophic bacterium Chloroflexus aurantiacus were investigated for the presence of lipopolysaccharide (LPS). With both strains, all fractions of hot phenol-water extracts and the extracted cell residues from whole cells or cell homogenates were found to be free from cha...

Meissner, J; Krauss, J H; Jürgens, U J; Weckesser, J

344

Tumor inhibition in mice by lipopolysaccharide-induced peritoneal cells and an induced soluble factor.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Lipopolysaccharide (LPS) was shown to prevent tumor growth in BALB/c mice when administered either prior to or after the inoculation of lethal doses of tumor cells. An attempt to elucidate the mechanism of this phenomenon utilizing in vivo protocols was made by the adoptive transfer of tumor protect...

Berendt, M J; Saluk, P H

345

Induction of autoimmunity in good and poor responder mice with mouse thyroglobulin and lipopolysaccharide  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The administration of soluble mouse thyroglobulin (MTg) in conjunction with bacterial lipopolysaccharide (LPS) led to the termination of natural tolerance to MTg in mice. The extent of autoimmunity correlated with responsiveness to MTg, previously shown by the injection of MTg in complete Freund's a...

346

Antibody Responses to Escherichia coli O157 and Other Lipopolysaccharides in Healthy Children and Adults  

Digital Repository Infrastructure Vision for European Research (DRIVER)

In Mexico, diarrheal disease due to different serotypes of Escherichia coli is highly prevalent, with only sporadic isolation of O157 non-H7 strains. This could be due to exposure to the O157 or related E. coli lipopolysaccharide (LPS), such as O7 or O116, at an early age. By using enzyme-linked imm...

Navarro, Armando; Eslava, Carlos; Hernandez, Ulises; Navarro-Henze, Jose Luis; Aviles, Magali; Garcia-de la Torre, Guadalupe

347

Cerium dioxide nanoparticles do not modulate the lipopolysaccharide-induced inflammatory response in human monocytes.  

UK PubMed Central (United Kingdom)

BACKGROUND: Cerium dioxide (CeO(2)) nanoparticles have potential therapeutic applications and are widely used for industrial purposes. However, the effects of these nanoparticles on primary human cells are largely unknown. The ability of nanoparticles to exacerbate pre-existing inflammatory disorders is not well documented for engineered nanoparticles, and is certainly lacking for CeO(2) nanoparticles. We investigated the inflammation-modulating effects of CeO(2) nanoparticles at noncytotoxic concentrations in human peripheral blood monocytes. METHODS: CD14(+) cells were isolated from peripheral blood samples of human volunteers. Cells were exposed to either 0.5 or 1 ?g/mL of CeO(2) nanoparticles over a period of 24 or 48 hours with or without lipopolysaccharide (10 ng/mL) prestimulation. Modulation of the inflammatory response was studied by measuring secreted tumor necrosis factor-alpha, interleukin-1beta, macrophage chemotactic protein-1, interferon-gamma, and interferon gamma-induced protein 10. RESULTS: CeO(2) nanoparticle suspensions were thoroughly characterized using dynamic light scattering analysis (194 nm hydrodynamic diameter), zeta potential analysis (-14 mV), and transmission electron microscopy (irregular-shaped particles). Transmission electron microscopy of CD14(+) cells exposed to CeO(2) nanoparticles revealed that these nanoparticles were efficiently internalized by monocytes and were found either in vesicles or free in the cytoplasm. However, no significant differences in secreted cytokine profiles were observed between CeO(2) nanoparticle-treated cells and control cells at noncytotoxic doses. No significant effects of CeO(2) nanoparticle exposure subsequent to lipopolysaccharide priming was observed on cytokine secretion. Moreover, no significant difference in lipopolysaccharide-induced cytokine production was observed after exposure to CeO(2) nanoparticles followed by lipopolysaccharide exposure. CONCLUSION: CeO(2) nanoparticles at noncytotoxic concentrations neither modulate pre-existing inflammation nor prime for subsequent exposure to lipopolysaccharides in human monocytes from healthy subjects.

Hussain S; Al-Nsour F; Rice AB; Marshburn J; Ji Z; Zink JI; Yingling B; Walker NJ; Garantziotis S

2012-01-01

348

Comparison of the limulus amebocyte lysate test and gas chromatography-mass spectrometry for measuring lipopolysaccharides (endotoxins) in airborne dust from poultry-processing industries.  

Science.gov (United States)

The lipopolysaccharide (endotoxin) content in airborne dust samples from three different poultry slaughterhouses was determined with both the chromogenic Limulus amebocyte lysate assay and gas chromatography-mass spectrometry analysis of lipopolysaccharide-derived 3-hydroxy fatty acids. Gram-negative cell walls were also measured by using two-dimensional gas chromatography/electron-capture analysis of diaminopimelic acid originating from the peptidoglycan. The correlation between the results of the Limulus assay and those of gas chromatography-mass spectrometry for determination of the lipopolysaccharide content in the dust samples was poor, whereas a good correlation was obtained between lipopolysaccharide and diaminopimelic acid concentrations with the gas chromatographic methods. The results suggest that it is predominantly cell-wall-dissociated lipopolysaccharides that are measured with the Limulus assay, whereas the gas chromatographic methods allow determination of total concentrations of lipopolysaccharide, including Limulus-inactive lipopolysaccharide, gram-negative cells, and cellular debris. PMID:2187411

Sonesson, A; Larsson, L; Schütz, A; Hagmar, L; Hallberg, T

1990-05-01

349

Comparison of the limulus amebocyte lysate test and gas chromatography-mass spectrometry for measuring lipopolysaccharides (endotoxins) in airborne dust from poultry-processing industries.  

UK PubMed Central (United Kingdom)

The lipopolysaccharide (endotoxin) content in airborne dust samples from three different poultry slaughterhouses was determined with both the chromogenic Limulus amebocyte lysate assay and gas chromatography-mass spectrometry analysis of lipopolysaccharide-derived 3-hydroxy fatty acids. Gram-negative cell walls were also measured by using two-dimensional gas chromatography/electron-capture analysis of diaminopimelic acid originating from the peptidoglycan. The correlation between the results of the Limulus assay and those of gas chromatography-mass spectrometry for determination of the lipopolysaccharide content in the dust samples was poor, whereas a good correlation was obtained between lipopolysaccharide and diaminopimelic acid concentrations with the gas chromatographic methods. The results suggest that it is predominantly cell-wall-dissociated lipopolysaccharides that are measured with the Limulus assay, whereas the gas chromatographic methods allow determination of total concentrations of lipopolysaccharide, including Limulus-inactive lipopolysaccharide, gram-negative cells, and cellular debris.

Sonesson A; Larsson L; Schütz A; Hagmar L; Hallberg T

1990-05-01

350

Comparative evaluation of two structurally related flavonoids, isoliquiritigenin and liquiritigenin, for their oral infection therapeutic potential.  

UK PubMed Central (United Kingdom)

Isoliquiritigenin (1) and liquiritigenin (2) are structurally related flavonoids found in a variety of plants. The purpose of this study was to perform a comparative analysis of biological properties of these compounds in regard to their therapeutic potential for oral infections. Compound 1 demonstrated significant antibacterial activity against three major periodontopathogens, Porphyromonas gingivalis, Fusobacterium nucleatum, and Prevotella intermedia. In contrast, 2 exerted less pronounced effects on the above bacterial species. Neither compound was effective against cariogenic bacteria (Streptococcus mutans and Streptococcus sobrinus). Furthermore, 1 exhibited a stronger inhibitory activity than 2 toward P. gingivalis collagenase and human matrix metalloproteinase 9. Finally, the capacity of 1 to attenuate the inflammatory response of macrophages induced by Aggregatibacter actinomycetemcomitans lipopolysaccharide (LPS) was much higher when compared to 2. The activation of transcriptional factors nuclear factor-?B (NF-?B) p65 and activator protein-1 (AP-1) associated with the LPS-induced inflammatory response in macrophages was inhibited strongly by 1, but less affected by 2.

Feldman M; Santos J; Grenier D

2011-09-01

351

Comparative evaluation of two structurally related flavonoids, isoliquiritigenin and liquiritigenin, for their oral infection therapeutic potential.  

Science.gov (United States)

Isoliquiritigenin (1) and liquiritigenin (2) are structurally related flavonoids found in a variety of plants. The purpose of this study was to perform a comparative analysis of biological properties of these compounds in regard to their therapeutic potential for oral infections. Compound 1 demonstrated significant antibacterial activity against three major periodontopathogens, Porphyromonas gingivalis, Fusobacterium nucleatum, and Prevotella intermedia. In contrast, 2 exerted less pronounced effects on the above bacterial species. Neither compound was effective against cariogenic bacteria (Streptococcus mutans and Streptococcus sobrinus). Furthermore, 1 exhibited a stronger inhibitory activity than 2 toward P. gingivalis collagenase and human matrix metalloproteinase 9. Finally, the capacity of 1 to attenuate the inflammatory response of macrophages induced by Aggregatibacter actinomycetemcomitans lipopolysaccharide (LPS) was much higher when compared to 2. The activation of transcriptional factors nuclear factor-?B (NF-?B) p65 and activator protein-1 (AP-1) associated with the LPS-induced inflammatory response in macrophages was inhibited strongly by 1, but less affected by 2. PMID:21866899

Feldman, Mark; Santos, Juliana; Grenier, Daniel

2011-08-25

352

Cerium dioxide nanoparticles do not modulate the lipopolysaccharide-induced inflammatory response in human monocytes  

Directory of Open Access Journals (Sweden)

Full Text Available Salik Hussain1,*, Faris Al-Nsour1,*, Annette B Rice1, Jamie Marshburn1, Zhaoxia Ji2, Jeffery I Zink2, Brenda Yingling1, Nigel J Walker3, Stavros Garantziotis11Clinical Research Unit, National Institute of Environmental Health Sciences/National Institute of Health, Research Triangle Park, NC, 2UC Center for Environmental Implications of Nanotechnology University of California, Los Angeles, CA, 3Division of National Toxicology Program, National Institute of Environmental Health Sciences/National Institute of Health, Research Triangle Park, NC, USA*Both are principal authorsBackground: Cerium dioxide (CeO2) nanoparticles have potential therapeutic applications