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Effect of anti-CD14 antibody on experimental periodontitis induced by Porphyromonas gingivalis lipopolysaccharide.  

Science.gov (United States)

The lipopolysaccharide (LPS) released by Porphyromonas gingivalis, a Gram-negative bacterium found in the periodontal pockets of patients with periodontitis, induces bone resorbing activity in vivo. We previously showed that a receptor for LPS on human gingival fibroblasts and gingival epithelial cells is CD14. In this study, we established a mouse model of experimental periodontitis by applying a P. gingivalis LPS solution to the buccal region of mice. P. gingivalis LPS-induced bone resorption and interleukin-6 production in the gingival tissues were significantly inhibited by pretreatment with anti-CD14 antibody for 5 weeks prior to LPS treatment. This result suggests that anti-CD14 antibody may be usable as a prototype for the development of drugs for the treatment of periodontal disease. PMID:12120761

Wang, Pao-Li; Oido-Mori, Mari; Fujii, Takeo; Kowashi, Yusuke; Kikuchi, Masanori; Suetsugu, Yasushi; Tanaka, Junzo; Azuma, Yasutaka; Shinohara, Mitsuko; Ohura, Kiyoshi

2002-06-01

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Luteolin and fisetin inhibit the effects of lipopolysaccharide obtained from Porphyromonas gingivalis in human gingival fibroblasts.  

Science.gov (United States)

Periodontitis is an inflammatory process of infectious origin that affects the gums and, in severe cases, destroys connective tissue, leading to loss of the dental organ. Gram-negative Porphyromonas gingivalis bacteria are recovered from patients with chronic periodontitis. The polysaccharide obtained from these bacteria induces the expression of interleukin (IL)-1 beta, tumor necrosis factor, and IL-6. Flavonoids are molecules that participate in the control of inflammatory processes. We studied the role of the flavonoids fisetin, luteolin, myricetin, and morin in inhibiting the activation of mitogen-activated protein kinase (MAPK) and AKT as well as their role in lipopolysaccharide (LPS)-induced cyclooxygenase-2 (COX-2) transcription. All four of these flavonoids were found to inhibit MAPK and AKT. Fisetin and luteolin blocked the activation of MAPK and AKT to levels below basal levels. All of these flavonoids also blocked LPS-mediated COX-2 expression. PMID:23054013

Gutiérrez-Venegas, Gloria; Contreras-Sánchez, Anabel

2013-01-01

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Porphyromonas gingivalis galE Is Involved in Lipopolysaccharide O-Antigen Synthesis and Biofilm Formation?  

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Porphyromonas gingivalis is a crucial component of complex plaque biofilms that form in the oral cavity, resulting in the progression of periodontal disease. To elucidate the mechanism of periodontal biofilm formation, we analyzed the involvement of several genes related to the synthesis of polysaccharides in P. gingivalis. Gene knockout P. gingivalis mutants were constructed by insertion of an ermF-ermAM cassette; among these mutants, the galE mutant showed some characteristic phenotypes inv...

Nakao, Ryoma; Senpuku, Hidenobu; Watanabe, Haruo

2006-01-01

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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  

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Full Text Available SciELO Colombia | Language: Spanish 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 condicion [...] es 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. 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 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.

2008-12-01

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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  

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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

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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 SciELO Colombia | Language: Spanish 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 condicion [...] es 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. 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 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.

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Identification of Porphyromonas gingivalis lipopolysaccharide-binding proteins in human saliva.  

Science.gov (United States)

Porphyromonas gingivalis causes periodontal diseases and its lipopolysaccharide (LPS) is considered as a major virulence factor responsible for pathogenesis. Since initial recognition of P. gingivalis LPS (Pg.LPS) in the oral cavity might be crucial for the host response, we identified Pg.LPS-binding proteins (Pg.LPS-BPs) using Pg.LPS-immobilized beads and a high-resolution mass spectrometry. LPS purified from P. gingivalis was conjugated onto N-hydroxysuccinimidyl-Sepharose(®) 4 Fast Flow beads. Notably, Pg.LPS-conjugated beads could stimulate Toll-like receptor 2 (TLR2) as determined by a TLR2-depdendent reporter expression system using CHO/CD14/TLR2. In addition, the Pg.LPS-conjugated beads induced the production of inflammatory mediators such as nitric oxide and interferon-gamma-inducible protein-10 in the macrophage cell-line, RAW 264.7. These results imply that Pg.LPS retained its immunological properties during the conjugation process. Then, the Pg.LPS-conjugated beads were mixed with a pool of saliva obtained from nine human subjects to capture Pg.LPS-BPs and molecular identities were determined by LTQ-Orbitrap hybrid fourier transform mass spectrometry. Pg.LPS-BPs captured at high frequencies included alpha-amylase, cystatin, prolactin-inducible protein, lysozyme C, immunoglobulin components, serum albumin, lipocalin-1, and submaxillary gland androgen-regulated protein 3B. These proteins are known to be involved in bacterial adhesion and colonization, anti-microbial functions or modulation of immune responses. PMID:21742380

Choi, Seulggie; Baik, Jung Eun; Jeon, Jun Ho; Cho, Kun; Seo, Deog-Gyu; Kum, Kee-Yeon; Yun, Cheol-Heui; Han, Seung Hyun

2011-09-01

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Purificación y caracterización de lipopolisacáridos de Eikenella corrodens 23834 y Porphyromonas gingivalis W83 / Purification and characterization of lipopolysaccharide from Eikenella corrodens 23834 and Porphyromonas gingivalis W83  

Scientific Electronic Library Online (English)

Full Text Available SciELO Colombia | Language: Spanish Abstract in spanish La purificación de lipopolisacáridos (LPS) o endotoxinas y su caracterización es un aspecto esencial para estudios que buscan aclarar el papel de estas biomoléculas de bacterias Gram negativas presentes en la cavidad oral y su relación con enfermedades locales periodontales y sistémicas. Este estudi [...] o implementa una metodología para la extracción, purificación y caracterización de LPS a partir de bacteria completa de Eikenella corrodens 23834 y Porphyromonas gingivalis W83, utilizando técnicas previamente descritas. La extracción cruda de LPS se realizó con fenol-agua caliente; la purificación se realizó con tratamiento enzimático con nucleasas y proteasa, seguido de cromatografía de exclusión por tamaño (Sephacryl S-200 HR) con deoxicolato de sodio como fase móvil. La caracterización de los extractos purificados se realizó por barrido espectrofotométrico, pruebas bioquímicas de electroforesis SDS-PAGE, ensayo Purpald y la prueba cromogénica de LAL. Como control para la identificación y caracterización de los extractos purificados se utilizaron LPS comerciales de Escherichia coli, Salmonella typhimurium, Rodobacter sphaeroides y Porphyromonas gingivalis. La metodología implementada permitió la obtención de LPS de elevada pureza con la identificación de KDO o heptosas, un quimiotipo de LPS-S (liso) para E. corrodens y LPS-SR (semi-rugoso) para P. gingivalis W83. Ambos LPS purificados mostraron capacidad endotóxica a bajas concentraciones. La metodología implementada en este estudio para la purificación y caracterización de LPS a partir de bacteria completa fue eficiente al compararla con los LPS comerciales. Abstract in english Purification of lipopolysaccharide (LPS) or endotoxins and its characterization is an important aspect for studies aimed at clarify the role of these biomolecules from Gram negative bacteria present in the oral cavity and its relationship with periodontal and systemic diseases. This study describes [...] an extraction, purification and characterization method of LPS from Eikenella corrodens 23834 and Porphyromonas gingivalis W83. LPS extraction was performed by using hot phenol-water; the purification was done with nuclease and protease enzymatic treatment, followed by size-exclusion chromatography (Sephacryl S-200 HR) with sodium deoxycholate as mobile phase. The characterization of the purified extracts was performed by spectrophotometric scanning, SDS-PAGE biochemical tests, Purpald assay and chromogenic LAL test. As control, commercial LPS from Escherichia coli, Salmonella typhimurium, P. gingivalis, and Rodobacter sphaeroides were used. The methodology mentioned above had allowed obtaining high purity LPS by identifying KDO or heptoses, a chemotype S-LPS (smooth) to E. corrodens; SR-LPS (semi-rough) for P. gingivalis W83. Both purified LPS showed endotoxic capacity at low concentrations. The methodology used in this study for purification and characterization of LPS from the whole bacteria was efficient when it was compared with commercial LPS.

Diego Fernando, Gualtero Escobar; Jeimy Paola, Porras Gaviria; Sebastian, Bernau Gutierrez; Diana Marcela, Buitrago Ramírez; Diana Marcela, Castillo Perdomo; Gloria Ines, Lafaurie Villamil.

2014-07-01

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Purificación y caracterización de lipopolisacáridos de Eikenella corrodens 23834 y Porphyromonas gingivalis W83 / Purification and characterization of lipopolysaccharide from Eikenella corrodens 23834 and Porphyromonas gingivalis W83  

Scientific Electronic Library Online (English)

Full Text Available SciELO Colombia | Language: Spanish Abstract in spanish La purificación de lipopolisacáridos (LPS) o endotoxinas y su caracterización es un aspecto esencial para estudios que buscan aclarar el papel de estas biomoléculas de bacterias Gram negativas presentes en la cavidad oral y su relación con enfermedades locales periodontales y sistémicas. Este estudi [...] o implementa una metodología para la extracción, purificación y caracterización de LPS a partir de bacteria completa de Eikenella corrodens 23834 y Porphyromonas gingivalis W83, utilizando técnicas previamente descritas. La extracción cruda de LPS se realizó con fenol-agua caliente; la purificación se realizó con tratamiento enzimático con nucleasas y proteasa, seguido de cromatografía de exclusión por tamaño (Sephacryl S-200 HR) con deoxicolato de sodio como fase móvil. La caracterización de los extractos purificados se realizó por barrido espectrofotométrico, pruebas bioquímicas de electroforesis SDS-PAGE, ensayo Purpald y la prueba cromogénica de LAL. Como control para la identificación y caracterización de los extractos purificados se utilizaron LPS comerciales de Escherichia coli, Salmonella typhimurium, Rodobacter sphaeroides y Porphyromonas gingivalis. La metodología implementada permitió la obtención de LPS de elevada pureza con la identificación de KDO o heptosas, un quimiotipo de LPS-S (liso) para E. corrodens y LPS-SR (semi-rugoso) para P. gingivalis W83. Ambos LPS purificados mostraron capacidad endotóxica a bajas concentraciones. La metodología implementada en este estudio para la purificación y caracterización de LPS a partir de bacteria completa fue eficiente al compararla con los LPS comerciales. Abstract in english Purification of lipopolysaccharide (LPS) or endotoxins and its characterization is an important aspect for studies aimed at clarify the role of these biomolecules from Gram negative bacteria present in the oral cavity and its relationship with periodontal and systemic diseases. This study describes [...] an extraction, purification and characterization method of LPS from Eikenella corrodens 23834 and Porphyromonas gingivalis W83. LPS extraction was performed by using hot phenol-water; the purification was done with nuclease and protease enzymatic treatment, followed by size-exclusion chromatography (Sephacryl S-200 HR) with sodium deoxycholate as mobile phase. The characterization of the purified extracts was performed by spectrophotometric scanning, SDS-PAGE biochemical tests, Purpald assay and chromogenic LAL test. As control, commercial LPS from Escherichia coli, Salmonella typhimurium, P. gingivalis, and Rodobacter sphaeroides were used. The methodology mentioned above had allowed obtaining high purity LPS by identifying KDO or heptoses, a chemotype S-LPS (smooth) to E. corrodens; SR-LPS (semi-rough) for P. gingivalis W83. Both purified LPS showed endotoxic capacity at low concentrations. The methodology used in this study for purification and characterization of LPS from the whole bacteria was efficient when it was compared with commercial LPS.

Diego Fernando, Gualtero Escobar; Jeimy Paola, Porras Gaviria; Sebastian, Bernau Gutierrez; Diana Marcela, Buitrago Ramírez; Diana Marcela, Castillo Perdomo; Gloria Ines, Lafaurie Villamil.

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Porphyromonas gingivalis lipopolysaccharide weakly activates M1 and M2 polarized mouse macrophages but induces inflammatory cytokines.  

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Porphyromonas gingivalis is associated with chronic periodontitis, an inflammatory disease of the tooth's supporting tissues. Macrophages are important in chronic inflammatory conditions, infiltrating tissue and becoming polarized to an M1 or M2 phenotype. As responses to stimuli differ between these phenotypes, we investigated the effect of P. gingivalis lipopolysaccharide (LPS) on M1 and M2 macrophages. M1 and M2 polarized macrophages were produced from murine bone marrow macrophages (BMM?) primed with gamma interferon (IFN-?) or interleukin-4 (IL-4), respectively, and incubated with a low or high dose of P. gingivalis LPS or control TLR2 and TLR4 ligands. In M1-M?, the high dose of P. gingivalis LPS (10 ?g/ml) significantly increased the expression of CD40, CD86, inducible nitric oxide synthase, and nitric oxide secretion. The low dose of P. gingivalis LPS (10 ng/ml) did not induce costimulatory or antibacterial molecules but did increase the secretion of IL-1?, IL-6, IL-12p40, IL-12p70, and tumor necrosis factor alpha (TNF-?). P. gingivalis LPS marginally increased the expression of CD206 and YM-1, but it did enhance arginase expression by M2-M?. Furthermore, the secretion of the chemokines KC, RANTES, eotaxin, and MCP-1 from M1, M2, and nonpolarized M? was enhanced by P. gingivalis LPS. TLR2/4 knockout macrophages combined with the TLR activation assays indicated that TLR2 is the main activating receptor for P. gingivalis LPS and whole cells. In conclusion, although P. gingivalis LPS weakly activated M1-M? or M2-M? compared to control TLR ligands, it induced the secretion of inflammatory cytokines, particularly TNF-? from M1-M? and IL-10 from M2-M?, as well as chemotactic chemokines from polarized macrophages. PMID:25047849

Holden, James A; Attard, Troy J; Laughton, Katrina M; Mansell, Ashley; O'Brien-Simpson, Neil M; Reynolds, Eric C

2014-10-01

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Lipopolysaccharides of Bacteroides intermedius (Prevotella intermedia) and Bacteroides (Porphyromonas) gingivalis induce interleukin-8 gene expression in human gingival fibroblast cultures.  

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Lipopolysaccharides (LPS) prepared from Bacteroides intermedius (Prevotella intermedia) and Bacteroides (Porphyromonas) gingivalis by hot phenol-water extraction induced interleukin-8 (IL-8) mRNA in normal human gingival fibroblast cultures, as demonstrated by Northern (RNA) blot analysis. IL-8 mRNA levels began to increase after a 2-h exposure, reached a maximum after 12 h, and then dropped to the unstimulated level at 48 h. IL-8 mRNA levels were also enhanced in a dose-dependent manner. By ...

Tamura, M.; Tokuda, M.; Nagaoka, S.; Takada, H.

1992-01-01

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Heme oxygenase-1 signals are involved in preferential inhibition of pro-inflammatory cytokine release by surfactin in cells activated with Porphyromonas gingivalis lipopolysaccharide.  

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Porphyromonas gingivalis is considered the major pathogen of periodontal disease, which leads to chronic inflammation in oral tissues. P. gingivalis-produced lipopolysaccharide (LPS) is a key factor in the development of periodontitis. It is established that surfactin produced by Bacillus subtilis confers anti-inflammatory properties. However, the underlying mechanisms responsible for surfactin-induced anti-inflammatory actions in the context of periodontitis are poorly understood. In this study, we investigated whether surfactin affected P. gingivalis LPS-induced pro-inflammatory cytokines, including tumor necrosis factor-?, interleukin (IL)-1?, IL-6, and IL-12, and determined that it significantly inhibited their production. Surfactin-mediated inhibition was mainly due to blocked activation of P. gingivalis LPS-triggered nuclear factor-?B. We also examined whether the regulatory effect of surfactin on P. gingivalis LPS-stimulated human THP-1 macrophages was mediated by the induction of heme oxygenase-1 (HO-1) signals, and determined that surfactin also induced HO-1 mRNA and protein expression via activation of Nrf-2. Additionally, we found that small interfering RNA-mediated knock-down of Nrf-2 significantly inhibited surfactin-induced HO-1 expression. Furthermore, inhibition of phosphoinositide 3-kinase (PI3K)/Akt and extracellular signal-regulated kinase (ERK) significantly decreased surfactin-induced HO-1 expression, which is consistent with the suggestion that surfactin-induced HO-1 expression occurs via PI3K/Akt, ERK, and Nrf-2. Treatment with a selective inhibitor of HO-1 reversed the surfactin-mediated inhibition of pro-inflammatory cytokines, suggesting that surfactin induces anti-inflammatory effects by activating Nrf-2-mediated HO-1 induction via PI3K/Akt and ERK signaling. Collectively, these observations support the potential of surfactin as a candidate in strategies to prevent caries, periodontitis, or other inflammatory diseases. PMID:20833156

Park, Sun Young; Kim, Young Hun; Kim, Eun-Kyoung; Ryu, Eun Yeon; Lee, Sang-Joon

2010-12-01

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Porphyromonas gingivalis: a clonal pathogen?  

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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

2011-11-01

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Gingipain aminopeptidase activities in Porphyromonas gingivalis  

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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. All 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 degradation of proteins and peptides P. gingivalis. PMID:23667904

Veillard, Florian; Potempa, Barbara; Poreba, Marcin; Drag, Marcin; Potempa, Jan

2014-01-01

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Characterization of the relA Gene of Porphyromonas gingivalis  

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Based upon the nucleotide sequence of the relA gene from Escherichia coli, a gene fragment corresponding to the homologous gene from the pathogenic oral bacterium Porphyromonas gingivalis 381 was isolated by PCR and utilized to construct a relA mutant. The mutant, KS7, was defective in ribosome-mediated ppGpp formation and also in the stringent response.

Sen, Keya; Hayashi, Jun-ichiro; Kuramitsu, Howard K.

2000-01-01

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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

2011-10-01

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Prevalence of Porphyromonas gingivalis four rag locus genotypes in patients of orthodontic gingivitis and periodontitis.  

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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 (pPorphyromonas gingivalis/rag locus and gingival index. rag-3 and rag-4 were the predominant genotypes in the patients of orthodontic gingivitis and mild-to-moderate periodontitis in Shandong. Porphyromonas.gingivalis carrying rag-1 has the strong virulence and could be associated with severe periodontitis. PMID:23593379

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

2013-01-01

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Comparative transcriptomic analysis of Porphyromonas gingivalis biofilm and planktonic cells  

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Full Text Available Abstract Background Porphyromonas gingivalis in subgingival dental plaque, as part of a mature biofilm, has been strongly implicated in the onset and progression of chronic periodontitis. In this study using DNA microarray we compared the global gene expression of a P. gingivalis biofilm with that of its planktonic counterpart grown in the same continuous culture. Results Approximately 18% (377 genes, at 1.5 fold or more, P-value P. gingivalis genome was differentially expressed when the bacterium was grown as a biofilm. Genes that were down-regulated in biofilm cells, relative to planktonic cells, included those involved in cell envelope biogenesis, DNA replication, energy production and biosynthesis of cofactors, prosthetic groups and carriers. A number of genes encoding transport and binding proteins were up-regulated in P. gingivalis biofilm cells. Several genes predicted to encode proteins involved in signal transduction and transcriptional regulation were differentially regulated and may be important in the regulation of biofilm growth. Conclusion This study analyzing global gene expression provides insight into the adaptive response of P. gingivalis to biofilm growth, in particular showing a down regulation of genes involved in growth and metabolic activity.

Lissel J Patricia

2009-01-01

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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 functions as a dominant receptor for P. gingivalis and contributes to P. gingivalis colonization.

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

2004-01-01

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Reducing medium for the cultivation of Porphyromonas gingivalis.  

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Porphyromonas gingivalis, a micro-organism frequently associated with human periodontal diseases, is commonly cultured under a reducing atmosphere in enclosed cabinets or glove boxes. This paper reports a modified Wilkins-Chalgren (MWC) medium for the culture of P. gingivalis under atmospheric conditions at 37 degrees C. On the basis of preliminary tests, WC broth was supplemented as follows: 500 mg/l cysteine hydrochloride; 250 mg/l sodium thioglycolate; and 1,000 mg/l sodium bicarbonate. Three P. gingivalis isolates (JKG-I, 33277 and A7436) showed very similar growth over 24 and 48 h periods when cultured both in WC medium in an anaerobic chamber and in the MWC medium. Culture of these isolates both in the anaerobic chamber and in the MWC medium yielded very similar data with respect to trypsin-like activity, total protease activity, and reactivity to monoclonal antibodies specific for P. gingivalis. Growth in the MWC medium varied over a 3- to 4-fold range for seven additional isolates (JKG-7, D86B6, D13B11, D84D2, JKG9, D67D9, D82F5) over 24 and 48 h periods. PMID:9237385

Pederson, E D; Turner, D W; Lamberts, B L; Schade, S Z

1997-01-01

 
 
 
 
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Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis in young Chinese adults.  

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The aim of this study was to determine the presence or absence of Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis in young Chinese adults and to examine the A. actinomycetemcomitans isolates from positive subjects with regard to the serotype distribution, presence of the leukotoxin gene lktA and the promoter for the leukotoxin operon as well as the incidence of phage Aa phi 23. Sixty subjects, working in a knitting factory in the Province of Guangzhou, People's Republic of China, were investigated. Subgingival microbial samples were taken from both upper first molars. They were cultured both anaerobically and in 5% CO2. P. gingivalis was found in 33 subjects. On average, it constituted 7% of the total anaerobic cultivable counts. A. actinomycetemcomitans was detected in 37 subjects of which seven yielded counts > 10(5). Twenty-one subjects were positive for both organisms. A. actinomycetemcomitans serotype a was found in 9 subjects, serotype c was found in 23 and serotype e in 5. A. actinomycetemcomitans serotypes b and d were not detected in any subjects. Presence of the leukotoxin gene lktA was demonstrated for all A. actinomycetemcomitans isolates; however, none of the A. actinomycetemcomitans strains from the present study had a deletion in the promoter region of the leukotoxin operon. The results of this investigation show a high frequency of the putative periodontal pathogens P. gingivalis and A. actinomycetemcomitans and corroborate the concept that there is variation in virulence and pathogenic potential among isolates from different subjects. PMID:10093538

Mombelli, A; Gmür, R; Frey, J; Meyer, J; Zee, K Y; Tam, J O; Lo, E C; Di Rienzo, J; Lang, N P; Corbet, E F

1998-08-01

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Identification of an O-antigen chain length regulator, WzzP, in Porphyromonas gingivalis  

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The periodontal pathogen Porphyromonas gingivalis has two different lipopolysaccharides (LPSs) designated O-LPS and A-LPS, which are a conventional O-antigen polysaccharide and an anionic polysaccharide that are both linked to lipid A-cores, respectively. However, the precise mechanisms of LPS biosynthesis remain to be determined. In this study, we isolated a pigment-less mutant by transposon mutagenesis and identified that the transposon was inserted into the coding sequence PGN_2005, which encodes a hypothetical protein of P. gingivalis ATCC 33277. We found that (i) LPSs purified from the PGN_2005 mutant were shorter than those of the wild type; (ii) the PGN_2005 protein was located in the inner membrane fraction; and (iii) the PGN_2005 gene conferred Wzz activity upon an Escherichia coli wzz mutant. These results indicate that the PGN_2005 protein, which was designated WzzP, is a functional homolog of the Wzz protein in P. gingivalis. Comparison of amino acid sequences among WzzP and conventional Wzz proteins indicated that WzzP had an additional fragment at the C-terminal region. In addition, we determined that the PGN_1896 and PGN_1233 proteins and the PGN_1033 protein appear to be WbaP homolog proteins and a Wzx homolog protein involved in LPS biosynthesis, respectively. PMID:23509024

Shoji, Mikio; Yukitake, Hideharu; Sato, Keiko; Shibata, Yasuko; Naito, Mariko; Aduse-Opoku, Joseph; Abiko, Yoshimitsu; Curtis, Michael A; Nakayama, Koji

2013-01-01

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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 extract e descritores clínicos periodontais. Material e Métodos: Indivíduos com periodontite (29) e com saúde periodontal (26) foram avaliados de acordo com as medidas de profundidade de son...

Trindade, Soraya Castro Et Al

2007-01-01

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Influence of resin coating materials on Porphyromonas gingivalis attachment.  

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Resin coating materials have been used for composite resin or provisional restoration in order to prevent plaque accumulation on their surfaces. However, it is not clear whether the coating materials influence attachment of periodontal bacteria. Therefore, we investigated the effect of resin coating materials on the attachment of Porphyromonas gingivalis (Pg). The polymerized auto cure resin plates were coated with two resin coating materials. To estimate the Pg attachment, each plate was immersed in brain heart infusion medium containing Pg. The quantity of bacteria attached on each plate was evaluated by crystal violet quantification. Morphological change of Pg was recorded using scanning electron microscopy. Both coating groups presented significantly lower Pg attachment compared to the control. The Pg shapes on the plates with resin coating materials were similar to the non-treated control plates. The resin coating materials clearly prevent Pg attachment on the polymerized auto cure resin plate. PMID:22277610

Kumada, Ai; Matsuka, Yoshizo; Mine, Atsushi; Ono, Mitsuaki; Uehara, Junji; Sonoi, Norihiro; Ito, Takashi; Takashiba, Shogo; Kuboki, Takuo

2012-02-01

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Characterization of Porphyromonas gingivalis Insertion Sequence-Like Element ISPg5  

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Porphyromonas gingivalis, a black-pigmented, gram-negative anaerobe, is found in periodontitis lesions, and its presence in subgingival plaque significantly increases the risk for periodontitis. In contrast to many bacterial pathogens, P. gingivalis strains display considerable variability, which is likely due to genetic exchange and intragenomic changes. To explore the latter possibility, we have studied the occurrence of insertion sequence (IS)-like elements in P. gingivalis W83 by utilizin...

Califano, Joseph V.; Kitten, Todd; Lewis, Janina P.; Macrina, Francis L.; Fleischmann, Robert D.; Fraser, Claire M.; Duncan, Margaret J.; Dewhirst, Floyd E.

2000-01-01

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Experimental Porphyromonas gingivalis infection in nonimmune athymic BALB/c mice.  

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The purpose of this report was to study the role of T lymphocytes following injection of Porphyromonas gingivalis in a mouse abscess model. Three invasive P. gingivalis isolates (ATCC 53977, W83, and AJW4) were injected into athymic BALB/c mice and their heterozygous (nu/+) littermates. The athymic BALB/c (nu/nu) mice were able to localize the invasive P. gingivalis isolates at the injection site. By comparison, the heterozygous BALB/c (nu/+) littermates developed hemorrhagic secondary lesion...

1991-01-01

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Asociación de la severidad de la periodontitis con niveles de cotinina y Porphyromonas gingivalis / Association between the severity of periodontitis with cotinine levels and Porphyromonas gingivalis  

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Full Text Available SciELO Cuba | Language: Spanish Abstract in spanish Fundamento: la cotinina aumenta los efectos de las toxinas producidas por los periodontopatógenos y se ha observado que el hábito de fumar altera la respuesta humoral e incrementa la infectividad de la Porphyromonas gingivalis. Objetivo: investigar la asociación entre los niveles de cotinina, la sev [...] eridad y la extensión de la periodontitis, entre los niveles de cotinina y presencia de P. gingivalis. Método: en el presente estudio de corte transversal, el universo estuvo constituido por 108 sujetos. Los parámetros periodontales se midieron en seis sitios por diente en todos los dientes, se excluyó el tercer molar. Se tomaron muestras de P. gingivalis en las bolsas periodontales. Resultados: al comparar fumadores y no fumadores se observaron diferencias estadísticamente significativas en la profundidad de sondaje y en el nivel de inserción clínica, con peores condiciones periodontales en los fumadores (p Abstract in english Background: cotinine increases the effects of the toxins produced by periodontopathogens and it has been observed that the smoking habit alters the humoral response and decreases the effectiveness of Porphyromonas gingivalis. Objective: to investigate the association between the cotinine levels and [...] the severity and extent of periodontitis; as well as between the cotinine levels and the presence of Porphyromonas gingivalis. Method: in the present cross-sectional study, the universe was composed of 108 individuals. The periodontal parameters were measured in six sites per tooth in all the teeth; the third molar was excluded. Some samples of Porphyromonas gingivalis in the periodontal pocket were taken. Results: when comparing smokers and non-smokers, differences statistically significant in the probing depth and in the clinical attachment level were observed with worse periodontal conditions in smokers (p

Carlos Martín, Ardila Medina; Isabel Cristina, Guzmán Zuluaga; Maria Adelaida, Vélez Echeverri.

2014-08-01

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Infection with Porphyromonas gingivalis Exacerbates Endothelial Injury in Obese Mice  

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Background A number of studies have revealed a link between chronic periodontitis and cardiovascular disease in obese patients. However, there is little information about the influence of periodontitis-associated bacteria, Porphyromonas gingivalis (Pg), on pathogenesis of atherosclerosis in obesity. Methods In vivo experiment: C57BL/6J mice were fed with a high-fat diet (HFD) or normal chow diet (CD), as a control. Pg was infected from the pulp chamber. At 6 weeks post-infection, histological and immunohistochemical analysis of aortal tissues was performed. In vitro experiment: hTERT-immortalized human umbilical vein endothelial cells (HuhT1) were used to assess the effect of Pg/Pg-LPS on free fatty acid (FFA) induced endothelial cells apoptosis and regulation of cytokine gene expression. Results Weaker staining of CD31 and increased numbers of TUNEL positive cells in aortal tissue of HFD mice indicated endothelial injury. Pg infection exacerbated the endothelial injury. Immunohistochemically, Pg was detected deep in the smooth muscle of the aorta, and the number of Pg cells in the aortal wall was higher in HFD mice than in CD mice. Moreover, in vitro, FFA treatment induced apoptosis in HuhT1 cells and exposure to Pg-LPS increased this effect. In addition, Pg and Pg-LPS both attenuated cytokine production in HuhT1 cells stimulated by palmitate. Conclusions Dental infection of Pg may contribute to pathogenesis of atherosclerosis by accelerating FFA-induced endothelial injury. PMID:25334003

Inubushi, Toshihiro; Kitagawa, Masae; Furusho, Hisako; Ando, Toshinori; Ayuningtyas, Nurina Febriyanti; Nagasaki, Atsuhiro; Ishihara, Kazuyuki; Tahara, Hidetoshi; Kozai, Katsuyuki; Takata, Takashi

2014-01-01

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Adhesion of Actinomyces viscosus to Porphyromonas (Bacteroides) gingivalis-coated hexadecane droplets.  

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Interbacterial adhesion (coadhesion) is considered a major determinant of dental plaque ecology. In this report, we studied several aspects of the adhesion of Porphyromonas (Bacteroides) gingivalis to hexadecane in order to use the liquid hydrocarbon as a convenient substratum for coadhesion assays. Washed suspensions of hydrophobic P. gingivalis 2561 cells were vortexed with hexadecane to yield highly stable cell-coated droplets. Kinetics of coadhesion between Actinomyces viscosus cells and ...

Rosenberg, M.; Buivids, I. A.; Ellen, R. P.

1991-01-01

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

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Previously, we have reported that gingipain activity in Porphyromonas gingivalis, the major causative agent in adult periodontitis, is post-translationally regulated by the unique Vim proteins including VimF, a putative glycosyltransferase. To further characterize VimF, an isogenic mutant defective in this gene in a different P. gingivalis genetic background was evaluated. In addition, the recombinant VimF protein was used to further confirm its glycosyltransferase function. The vimF-defectiv...

Muthiah, Arun S.; Aruni, Wilson; Robles, Antonette G.; Dou, Yuetan; Roy, Francis; Fletcher, Hansel M.

2013-01-01

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Effect of simulated high-altitude hypoxia on Porphyromonas gingivalis  

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Full Text Available Objective?To investigate the effects of simulated high-altitude hypoxia on the detection rate and endotoxin level of Porphyromonas gingivalis (Pg of subgingival bacterial plagues in rabbit periodontitis models. Methods?Forty male rabbits were randomly divided into four groups, namely, normoxia control group (group A1, normoxia experimental group (group A2, hypoxia control group (group B1, and hypoxia experimental group (group B2. Each group included 10 rabbits. Periodontitis models was established in groups A2 and B2 combined by ligating both lower central incisors with steel ligature and feeding periodontitis diets, and then the animals were housed in a hypoxia chamber (simulating 5000m altitude, 23h per day. Groups A1 and A2 were raised normal diet in normoxia environment. After eight weeks, the rabbit periodontitis model was evaluated by observing radiographic features of the X-ray films and histopathologic changes under a light microscope. Subgingival plague sample from periodontal pockets on both lower central incisors were collected for isolation, culture and identification of Pg, and for detection of the endotoxin level. Results?The histopathologic observation and X-ray examination results showed that the periodontitis of rabbits in group B2 was significantly more severe than that in group A2. The detection rates of Pg in group A1, A2, B1 and B2 was 0%, 50%, 55% and 95% (P < 0.05. Pg detection rate and endotoxin level were higher in group B2 (95%, 0.46±0.04EU/ml than in group A2 (50%, 0.38±0.02EU/ml, P < 0.05. Conclusions?The process speed and damage degree of periodontitis in hypoxic environment is higher than that in normoxic environment. Moreover, the hypoxic environment is more suitable in the colonization of Pg with higher endotoxin level in subgingival plague.

Jing-jing HUANG

2012-04-01

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NOD2 Contributes to Porphyromonas gingivalis-induced Bone Resorption.  

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The NOD-like receptors are cytoplasmic proteins that sense microbial by-products released by invasive bacteria. Although NOD1 and NOD2 are functionally expressed in cells from oral tissues and play a role triggering immune responses, the role of NOD2 receptor in the bone resorption and in the modulation of osteoclastogenesis is still unclear. We show that in an experimental model of periodontitis with Porphyromonas gingivalis W83, NOD2(-/-) mice showed lower bone resorption when compared to wild type. Quantitative polymerase chain reaction analysis revealed that wild-type infected mice showed an elevated RANKL/OPG ratio when compared to NOD2(-/-) infected mice. Moreover, the expression of 2 osteoclast activity markers-cathepsin K and matrix metalloproteinase 9-was significantly lower in gingival tissue from NOD2(-/-) infected mice compared to WT infected ones. The in vitro study reported an increase in the expression of the NOD2 receptor 24 hr after stimulation of hematopoietic bone marrow cells with M-CSF and RANKL. We also evaluated the effect of direct activation of NOD2 receptor on osteoclastogenesis, by the activation of this receptor in preosteoclasts culture, with different concentrations of muramyl dipeptide. The results show no difference in the number of TRAP-positive cells. Although it did not alter the osteoclasts differentiation, the activation of NOD2 receptor led to a significant increase of cathepsin K expression. We confirm that this enzyme was active, since the osteoclasts resorption capacity was enhanced by muramyl dipeptide stimulation, evaluated in osteoassay plate. These results show that the lack of NOD2 receptor impairs the bone resorption, suggesting that NOD2 receptor could contribute to the progression of bone resorption in experimental model of periodontitis. The stimulation of NOD2 by its agonist, muramyl dipeptide, did not affect osteoclastogenesis, but it does favor the bone resorption capacity identified by increased osteoclast activity. PMID:25239844

Prates, T P; Taira, T M; Holanda, M C; Bignardi, L A; Salvador, S L; Zamboni, D S; Cunha, F Q; Fukada, S Y

2014-11-01

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Porphyromonas gingivalis: keeping the pathos out of the biont  

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Full Text Available The primary goal of the human microbiome initiative has been to increase our understanding of the structure and function of our indigenous microbiota and their effects on human health and predisposition to disease. Because of its clinical importance and accessibility for in vivo study, the oral biofilm is one of the best-understood microbial communities associated with the human body. Studies have shown that there is a succession of select microbial interactions that directs the maturation of a defined community structure, generating the formation of dental plaque. Although the initiating factors that lead to disease development are not clearly defined, in many individuals there is a fundamental shift from a health-associated biofilm community to one that is pathogenic in nature and a central player in the pathogenic potential of this community is the presence of Porphyromonas gingivalis. This anaerobic bacterium is a natural member of the oral microbiome, yet it can become highly destructive (termed pathobiont and proliferate to high cell numbers in periodontal lesions, which is attributed to its arsenal of specialized virulence factors. Hence, this organism is regarded as a primary etiologic agent of periodontal disease progression. In this review, we summarize some of the latest information regarding what is known about its role in periodontitis, including pathogenic potential as well as ecological and nutritional parameters that may shift this commensal to a virulent state. We also discuss parallels between the development of pathogenic biofilms and the human cellular communities that lead to cancer, specifically we frame our viewpoint in the context of ‘wounds that fail to heal’.

Carla Cugini

2013-04-01

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Characterization of a bacterial tyrosine kinase in Porphyromonas gingivalis involved in polymicrobial synergy.  

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Interspecies communication between Porphyromonas gingivalis and Streptococcus gordonii underlies the development of synergistic dual species communities. Contact with S. gordonii initiates signal transduction within P. gingivalis that is based on protein tyrosine (de)phosphorylation. In this study, we characterize a bacterial tyrosine (BY) kinase (designated Ptk1) of P. gingivalis and demonstrate its involvement in interspecies signaling. Ptk1 can utilize ATP for autophosphorylation and is dephosphorylated by the P. gingivalis tyrosine phosphatase, Ltp1. Community development with S. gordonii is severely abrogated in a ptk1 mutant of P. gingivalis, indicating that tyrosine kinase activity is required for maximal polymicrobial synergy. Ptk1 controls the levels of the transcriptional regulator CdhR and the fimbrial adhesin Mfa1 which mediates binding to S. gordonii. The ptk1 gene is in an operon with two genes involved in exopolysaccharide synthesis, and similar to other BY kinases, Ptk1 is necessary for exopolysaccharide production in P. gingivalis. Ptk1 can phosphorylate the capsule related proteins PGN_0224, a UDP-acetyl-mannosamine dehydrogenase, and PGN_0613, a UDP-glucose dehydrogenase, in P. gingivalis. Knockout of ptk1 in an encapsulated strain of P. gingivalis resulted in loss of capsule production. Collectively these results demonstrate that the P. gingivalis Ptk1 BY kinase regulates interspecies communication and controls heterotypic community development with S. gordonii through adjusting the levels of the Mfa1 adhesin and exopolysaccharide. PMID:24811194

Wright, Christopher J; Xue, Peng; Hirano, Takanori; Liu, Chengcheng; Whitmore, Sarah E; Hackett, Murray; Lamont, Richard J

2014-06-01

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Effects of Streptococcus thermophilus on volatile sulfur compounds produced by Porphyromonas gingivalis.  

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Halitosis as oral malodour is an unpleasant odour caused by volatile sulfur compounds (VSCs). VSCs are produced primarily by anaerobic bacteria that abundantly produce proteinase as trypsin-like enzyme. General therapies, such as mouthwash and plaque control, do not provide a continuous effect on oral halitosis. Streptococcus thermophilus is a probiotic bacterium that is beneficial for human health. The aim of this study was to evaluate the effect of S. thermophilus on Porphyromonas gingivalis-producing VSCs and to analyze the inhibitory mechanism of halitosis. P. gingivalis was cultured with or without S. thermophilus, and the emission of VSCs from the spent culture medium was measured by gas chromatography. In order to analyze the inhibitory effect, the antibacterial activity of S. thermophilus against P. gingivalis was assessed. After the spent culture medium or whole bacterial of S. thermophilus was mixed with the spent culture medium of P. gingivalis, VSCs were again measured by gas chromatograph. When S. thermophilus and P. gingivalis were co-cultivated, VSCs were present at a lower level than those of single-cultured P. gingivalis. S. thermophilus inhibited growth of P. gingivalis, and the whole bacteria and the spent culture medium of S. thermophilus reduced emission of VSCs gas. S. thermophilus may reduce oral malodour by inhibition of P. gingivalis growth and neutralizing VSCs with their metabolites or themselves. PMID:25105253

Lee, Sung-Hoon; Baek, Dong-Heon

2014-11-01

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VimA-Dependent Modulation of Acetyl Coenzyme A Levels and Lipid A Biosynthesis Can Alter Virulence in Porphyromonas gingivalis  

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The Porphyromonas gingivalis VimA protein has multifunctional properties that can modulate several of its major virulence factors. To further characterize VimA, P. gingivalis FLL406 carrying an additional vimA gene and a vimA-defective mutant in a different P. gingivalis genetic background were evaluated. The vimA-defective mutant (FLL451) in the P. gingivalis ATCC 33277 genetic background showed a phenotype similar to that of the vimA-defective mutant (FLL92) in the P. gingivalis W83 genetic...

Aruni, A. Wilson; Lee, J.; Osbourne, D.; Dou, Y.; Roy, F.; Muthiah, A.; Boskovic, D. S.; Fletcher, H. M.

2012-01-01

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Oral streptococcal glyceraldehyde-3-phosphate dehydrogenase mediates interaction with Porphyromonas gingivalis fimbriae.  

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Interaction of Porphyromonas gingivalis with plaque-forming bacteria is necessary for its colonization in periodontal pockets. Participation of Streptococcus oralis glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and P. gingivalis fimbriae in this interaction has been reported. In this investigation, the contribution of various oral streptococcal GAPDHs to interaction with P. gingivalis fimbriae was examined. Streptococcal cell surface GAPDH activity was measured by incubation of a constant number of streptococci with glyceraldehyde-3-phosphate and analysis for the conversion of NAD+ to NADH based on the absorbance at 340 nm. Coaggregation activity was measured by a turbidimetric assay. Cell surface GAPDH activity was correlated with coaggregation activity (r = 0.854, P mutanolysin and purified on a Cibacron Blue Sepharose column. Subsequently, their DNA sequences were elucidated. Purified GAPDHs bound P. gingivalis recombinant fimbrillin by Western blot assay, furthermore, their DNA sequences displayed a high degree of homology with one another. Moreover, S. oralis recombinant GAPDH inhibited coaggregation between P. gingivalis and the aforementioned five streptococcal strains in a dose-dependent manner. These results suggest that GAPDHs of various plaque-forming streptococci may be involved in their attachment to P. gingivalis fimbriae and that they may contribute to P. gingivalis colonization. PMID:15488735

Maeda, Kazuhiko; Nagata, Hideki; Nonaka, Aya; Kataoka, Kosuke; Tanaka, Muneo; Shizukuishi, Satoshi

2004-11-01

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

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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. PMID:23620122

Cai, Yu; Kurita-Ochiai, Tomoko; Hashizume, Tomomi; Yamamoto, Masafumi

2013-02-01

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Porphyromonas gingivalis Mediates Inflammasome Repression in Polymicrobial Cultures through a Novel Mechanism Involving Reduced Endocytosis*  

Science.gov (United States)

The interleukin (IL)-1?-processing inflammasome has recently been identified as a target for pathogenic evasion of the inflammatory response by a number of bacteria and viruses. We postulated that the periodontal pathogen, Porphyromonas gingivalis may suppress the inflammasome as a mechanism for its low immunogenicity and pathogenic synergy with other, more highly immunogenic periodontal bacteria. Our results show that P. gingivalis lacks signaling capability for the activation of the inflammasome in mouse macrophages. Furthermore, P. gingivalis can suppress inflammasome activation by another periodontal bacterium, Fusobacterium nucleatum. This repression affects IL-1? processing, as well as other inflammasome-mediated processes, including IL-18 processing and cell death, in both human and mouse macrophages. F. nucleatum activates IL-1? processing through the Nlrp3 inflammasome; however, P. gingivalis repression is not mediated through reduced levels of inflammasome components. P. gingivalis can repress Nlrp3 inflammasome activation by Escherichia coli, and by danger-associated molecular patterns and pattern-associated molecular patterns that mediate activation through endocytosis. However, P. gingivalis does not suppress Nlrp3 inflammasome activation by ATP or nigericin. This suggests that P. gingivalis may preferentially suppress endocytic pathways toward inflammasome activation. To directly test whether P. gingivalis infection affects endocytosis, we assessed the uptake of fluorescent particles in the presence or absence of P. gingivalis. Our results show that P. gingivalis limits both the number of cells taking up beads and the number of beads taken up for bead-positive cells. These results provide a novel mechanism of pathogen-mediated inflammasome inhibition through the suppression of endocytosis. PMID:22843689

Taxman, Debra J.; Swanson, Karen V.; Broglie, Peter M.; Wen, Haitao; Holley-Guthrie, Elizabeth; Huang, Max Tze-Han; Callaway, Justin B.; Eitas, Tim K.; Duncan, Joseph A.; Ting, Jenny P. Y.

2012-01-01

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Porphyromonas gingivalis mediates inflammasome repression in polymicrobial cultures through a novel mechanism involving reduced endocytosis.  

Science.gov (United States)

The interleukin (IL)-1?-processing inflammasome has recently been identified as a target for pathogenic evasion of the inflammatory response by a number of bacteria and viruses. We postulated that the periodontal pathogen, Porphyromonas gingivalis may suppress the inflammasome as a mechanism for its low immunogenicity and pathogenic synergy with other, more highly immunogenic periodontal bacteria. Our results show that P. gingivalis lacks signaling capability for the activation of the inflammasome in mouse macrophages. Furthermore, P. gingivalis can suppress inflammasome activation by another periodontal bacterium, Fusobacterium nucleatum. This repression affects IL-1? processing, as well as other inflammasome-mediated processes, including IL-18 processing and cell death, in both human and mouse macrophages. F. nucleatum activates IL-1? processing through the Nlrp3 inflammasome; however, P. gingivalis repression is not mediated through reduced levels of inflammasome components. P. gingivalis can repress Nlrp3 inflammasome activation by Escherichia coli, and by danger-associated molecular patterns and pattern-associated molecular patterns that mediate activation through endocytosis. However, P. gingivalis does not suppress Nlrp3 inflammasome activation by ATP or nigericin. This suggests that P. gingivalis may preferentially suppress endocytic pathways toward inflammasome activation. To directly test whether P. gingivalis infection affects endocytosis, we assessed the uptake of fluorescent particles in the presence or absence of P. gingivalis. Our results show that P. gingivalis limits both the number of cells taking up beads and the number of beads taken up for bead-positive cells. These results provide a novel mechanism of pathogen-mediated inflammasome inhibition through the suppression of endocytosis. PMID:22843689

Taxman, Debra J; Swanson, Karen V; Broglie, Peter M; Wen, Haitao; Holley-Guthrie, Elizabeth; Huang, Max Tze-Han; Callaway, Justin B; Eitas, Tim K; Duncan, Joseph A; Ting, Jenny P Y

2012-09-21

 
 
 
 
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In vitro models of tissue penetration and destruction by Porphyromonas gingivalis.  

Science.gov (United States)

Porphyromonas gingivalis is a gram-negative anaerobic bacterium that is considered the key etiologic agent of chronic periodontitis. Arg- and Lys-gingipain cysteine proteinases produced by P. gingivalis are key virulence factors and are believed to be essential for significant tissue component degradation, leading to host tissue invasion by periodontopathogens. Two in vitro models were used to determine the extent to which P. gingivalis can reach connective tissue. The tissue penetration potential of P. gingivalis was first investigated by using an engineered human oral mucosa model composed of normal human epithelial cells and fibroblasts. Internalized bacteria were assessed by transmission electron microscopy. Bacteria were observed within multilayered gingival epithelial cells and in the space between the stratified epithelium and the lamina propria. A gingipain-null mutant strain of P. gingivalis was found to be less potent in penetrating tissue than the wild-type strain. Proinflammatory responses to P. gingivalis infection were evaluated. P. gingivalis increased the secretion of interleukin-1 beta, interleukin-6, interleukin-8, and tumor necrosis factor alpha. In the second part of the study, the contribution of P. gingivalis gingipains to tissue penetration was investigated by using a reconstituted basement membrane model (Matrigel). The penetration of (14)C-labeled P. gingivalis cells through Matrigel was significantly reduced when leupeptin, a specific inhibitor of Arg-gingipain activity, was added or when a gingipain-null mutant was used. The results obtained with these two relevant models support the capacities of P. gingivalis to infiltrate periodontal tissue and to modulate the proinflammatory response and suggest a critical role of gingipains in tissue destruction. PMID:15271930

Andrian, Elisoa; Grenier, Daniel; Rouabhia, Mahmoud

2004-08-01

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Role of the Clp System in Stress Tolerance, Biofilm Formation, and Intracellular Invasion in Porphyromonas gingivalis?  

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Clp proteases and chaperones are ubiquitous among prokaryotes and eukaryotes, and in many pathogenic bacteria the Clp stress response system is also involved in regulation of virulence properties. In this study, the roles of ClpB, ClpC, and ClpXP in stress resistance, homotypic and heterotypic biofilm formation, and intracellular invasion in the oral opportunistic pathogen Porphyromonas gingivalis were investigated. Absence of ClpC and ClpXP, but not ClpB, resulted in diminished tolerance to ...

Capestany, Cindy A.; Tribble, Gena D.; Maeda, Kazuhiko; Demuth, Donald R.; Lamont, Richard J.

2008-01-01

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Inactivation of Membrane Tumor Necrosis Factor Alpha by Gingipains from Porphyromonas gingivalis  

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Gingipains are cysteine proteinases produced by Porphyromonas gingivalis, a major causative bacterium of adult periodontitis. They consist of arginine-specific (HRgpA and RgpB) and lysine-specific (Kgp) proteinases. Gingipains strongly affect the host defense system by degrading some cytokines, components of the complement system, and several immune cell receptors. In an in vitro model, gingipains were shown to degrade soluble tumor necrosis factor alpha (TNF-?). However, since membrane TNF-...

Me?z?yk-kopec?, Renata; Bzowska, Ma?gorzata; Potempa, Jan; Bzowska, Monika; Jura, Natalia; Sroka, Aneta; Black, Roy A.; Bereta, Joanna

2005-01-01

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Inhibition of Trypsin-Like Cysteine Proteinases (Gingipains) from Porphyromonas gingivalis by Tetracycline and Its Analogues  

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Extracellular cysteine proteinases, referred to as gingipains, are considered important virulence factors for Porphyromonas gingivalis, a bacterium recognized as a major etiologic agent of chronic periodontitis. We investigated the effect of tetracycline and its analogues, doxycycline and minocycline, on the enzymatic activities of gingipains. Tetracyclines at 100 ?M totally inhibited the amidolytic activity of arginine-specific gingipains (HRgpA and RgpB). In contrast, inhibition of Kgp was...

Imamura, Takahisa; Matsushita, Kenji; Travis, James; Potempa, Jan

2001-01-01

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Porphyromonas gingivalis induce apoptosis in human gingival epithelial cells through a gingipain-dependent mechanism  

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Full Text Available Abstract Background The oral pathogen Porphyromonas gingivalis has been shown to modulate apoptosis in different cell types, but its effect on epithelial cells remains unclear. Results We demonstrate that primary human gingival epithelial cells (HGECs challenged with live P. gingivalis for 24 hours exhibit apoptosis, and we characterize this by M30 epitope detection, caspase-3 activity, DNA fragmentation and Annexin-V staining. Live bacteria strongly upregulated intrinsic and extrinsic apoptotic pathways. Pro-apoptotic molecules such as caspase-3, -8, -9, Bid and Bax were upregulated after 24 hours. The anti-apoptotic Bcl-2 was also upregulated, but this was not sufficient to ensure cell survival. The main P. gingivalis proteases arginine and lysine gingipains are necessary and sufficient to induce host cell apoptosis. Thus, live P. gingivalis can invoke gingival epithelial cell apoptosis in a time and dose dependent manner with significant apoptosis occurring between 12 and 24 hours of challenge via a gingipain-dependent mechanism. Conclusion The present study provides evidence that live, but not heat-killed, P. gingivalis can induce apoptosis after 24 hours of challenge in primary human gingival epithelial cells. Either arginine or lysine gingipains are necessary and sufficient factors in P. gingivalis elicited apoptosis.

Garcia Carlos A

2009-05-01

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Intercellular spreading of Porphyromonas gingivalis infection in primary gingival epithelial cells.  

Science.gov (United States)

Porphyromonas gingivalis, an important periodontal pathogen, is an effective colonizer of oral tissues. The organism successfully invades, multiplies in, and survives for extended periods in primary gingival epithelial cells (GECs). It is unknown whether P. gingivalis resides in the cytoplasm of infected cells throughout the infection or can spread to adjacent cells over time. We developed a technique based on flow cytofluorometry and fluorescence microscopy to study propagation of the organism at different stages of infection of GECs. Results showed that P. gingivalis spreads cell to cell and that the amount of spreading increases gradually over time. There was a very low level of propagation of bacteria to uninfected cells early in the infection (3 h postinfection), but there were 20-fold and 45-fold increases in the propagation rate after 24 h and 48 h, respectively, of infection. Immunofluorescence microscopy of infected cells suggested that intercellular translocation of P. gingivalis may be mediated through actin-based membrane protrusions, bypassing the need for release of bacteria into extracellular medium. Consistent with these observations, cytochalasin D treatment of infected cells resulted in significant inhibition of bacterial spreading. This study shows for the first time that P. gingivalis disseminates from cell to cell without passing through the extracellular space. This mechanism of spreading may allow P. gingivalis to colonize oral tissues without exposure to the humoral immune response. PMID:16369027

Yilmaz, Ozlem; Verbeke, Philippe; Lamont, Richard J; Ojcius, David M

2006-01-01

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Susceptibility of Porphyromonas gingivalis and Streptococcus mutans to Antibacterial Effect from Mammea americana.  

Science.gov (United States)

The development of periodontal disease and dental caries is influenced by several factors, such as microorganisms of bacterial biofilm or commensal bacteria in the mouth. These microorganisms trigger inflammatory and immune responses in the host. Currently, medicinal plants are treatment options for these oral diseases. Mammea americana extracts have reported antimicrobial effects against several microorganisms. Nevertheless, this effect is unknown against oral bacteria. Therefore, the aim of this study was to evaluate the antibacterial effect of M. americana extract against Porphyromonas gingivalis and Streptococcus mutans. For this, an experimental study was conducted. Ethanolic extract was obtained from seeds of M. americana (one oil phase and one ethanolic phase). The strains of Porphyromonas gingivalis ATCC 33277 and Streptococcus mutans ATCC 25175 were exposed to this extract to evaluate its antibacterial effect. Antibacterial activity was observed with the two phases of M. americana extract on P. gingivalis and S. mutans with lower MICs (minimum inhibitory concentration). Also, bactericidal and bacteriostatic activity was detected against S. mutans, depending on the concentration of the extract, while on M. americana extract presented only bacteriostatic activity against P. gingivalis. These findings provide important and promising information allowing for further exploration in the future. PMID:24864137

Herrera Herrera, Alejandra; Franco Ospina, Luis; Fang, Luis; Díaz Caballero, Antonio

2014-01-01

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

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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

2007-01-01

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Development of a novel plasmid vector pTIO-1 adapted for electrotransformation of Porphyromonas gingivalis.  

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We report here the construction of a plasmid vector designed for the efficient electrotransformation of the periodontal pathogen Porphyromonas gingivalis. The novel Escherichia coli-Bacteroides/P. gingivalis shuttle vector, designated pTIO-1, is based on the 11.0-kb E. coli-Bacteroides conjugative shuttle vector, pVAL-1 (a pB8-51 derivative). To construct pTIO-1, the pB8-51 origin of replication and erythromycin resistance determinant of pVAL-1 were cloned into the E. coli cloning vector pBluescript II SK(-) and non-functional regions were deleted. pTIO-1 has an almost complete multiple cloning site from pBluescript II SK(-). The size of pTIO-1 is 4.5kb, which is convenient for routine gene manipulation. pTIO-1 was introduced into P. gingivalis via electroporation, and erythromycin-resistant transformants carrying pTIO-1 were obtained. We characterized the transformation efficiency, copy number, host range, stability, and insert size capacity of pTIO-1. An efficient plasmid electrotransformation of P. gingivalis will facilitate functional analysis and expression of P. gingivalis genes, including the virulence factors of this bacterium. PMID:25102110

Tagawa, Junpei; Inoue, Tetsuyoshi; Naito, Mariko; Sato, Keiko; Kuwahara, Tomomi; Nakayama, Masaaki; Nakayama, Koji; Yamashiro, Takashi; Ohara, Naoya

2014-10-01

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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

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Macrophage depletion abates Porphyromonas gingivalis-induced alveolar bone resorption in mice.  

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The role of the macrophage in the immunopathology of periodontitis has not been well defined. In this study, we show that intraoral inoculation of mice with Porphyromonas gingivalis resulted in infection, alveolar bone resorption, and a significant increase in F4/80(+) macrophages in gingival and submandibular lymph node tissues. Macrophage depletion using clodronate-liposomes resulted in a significant reduction in F4/80(+) macrophage infiltration of gingival and submandibular lymph node tissues and significantly (p CD206(+)), were the dominant macrophage phenotype of the gingival infiltrate in response to P. gingivalis infection. P. gingivalis induced a significant (p < 0.01) increase in NO production and a small increase in urea concentration, as well as a significant increase in the secretion of IL-1?, IL-6, IL-10, IL-12 (p70), eotaxin, G-CSF, GM-CSF, macrophage chemoattractant protein-1, macrophage inflammatory protein-? and -?, and TNF-? in isolated murine macrophages. In conclusion, P. gingivalis infection induced infiltration of functional/inflammatory M1 macrophages into gingival tissue and alveolar bone resorption. Macrophage depletion reduced P. gingivalis infection and alveolar bone resorption by modulating the host immune response. PMID:25070844

Lam, Roselind S; O'Brien-Simpson, Neil M; Lenzo, Jason C; Holden, James A; Brammar, Gail C; Walsh, Katrina A; McNaughtan, Judith E; Rowler, Dennis K; Van Rooijen, Nico; Reynolds, Eric C

2014-09-01

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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 SciELO Chile | Language: Spanish 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 tre [...] s 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. 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 informat [...] ion 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.

2012-12-01

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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 SciELO Colombia | Language: English 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 los 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 FimBr [...] iline 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 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.

Sandra, Moreno; Adolfo, Contreras.

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

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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

2010-04-01

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

Scientific Electronic Library Online (English)

Full Text Available SciELO Spain | Language: Spanish Abstract in spanish 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, s [...] e 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. Abstract in english 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 tha [...] t 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.

56

Identification and sequence analysis of a methylase gene in Porphyromonas gingivalis.  

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A gene from the periodontal organism Porphyromonas gingivalis has been identified as encoding a DNA methylase. The gene, referred to as pgiIM, has been sequenced and found to contain a reading frame of 864 basepairs. The putative amino acid sequence of the encoded methylase was 288 amino acids, and shared 47% and 31% homology with the Streptococcus pneumoniae DpnII and E. coli Dam methylases, respectively. The activity and specificity of the pgi methylase (M.PgiI) was confirmed by cloning the...

Banas, J. A.; Ferretti, J. J.; Progulske-fox, A.

1991-01-01

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Synthesis of sulfonamides with effective inhibitory action against Porphyromonas gingivalis ?-carbonic anhydrase.  

Science.gov (United States)

New benzenesulfonamides incorporating GABA or N-?-acetyl-L-lysine scaffolds as well as guanidine functionalities as water solubilizing moieties were obtained, using 4-aminoethyl/methyl-benzenesulfonamide and metanilamide/sulfanilamide as zinc-binding motives. The new compounds were medium potency inhibitors of the widespread cytosolic human carbonic anhydrase (CA, EC 4.2.1.1) isoforms I and II and more effective inhibitors (KIs low nanomolar range) of the bacterial ?-CA from the oral pathogen Porphyromonas gingivalis. These sulfonamides may be useful tools for understanding the physiological role of bacterial CAs in pathogenesis of some infectious disease. PMID:25011913

Ceruso, Mariangela; Del Prete, Sonia; AlOthman, Zeid; Osman, Sameh M; Scozzafava, Andrea; Capasso, Clemente; Supuran, Claudiu T

2014-08-15

58

FimR and FimS: biofilm formation and gene expression in Porphyromonas gingivalis.  

Science.gov (United States)

Porphyromonas gingivalis is a late-colonizing bacterium of the subgingival dental plaque biofilm associated with periodontitis. Two P. gingivalis genes, fimR and fimS, are predicted to encode a two-component signal transduction system comprising a response regulator (FimR) and a sensor histidine kinase (FimS). In this study, we show that fimS and fimR, although contiguous on the genome, are not part of an operon. We inactivated fimR and fimS in both the afimbriated strain W50 and the fimbriated strain ATCC 33277 and demonstrated that both mutants formed significantly less biofilm than their respective wild-type strains. Quantitative reverse transcription-real-time PCR showed that expression of fimbriation genes was reduced in both the fimS and fimR mutants of strain ATCC 33277. The mutations had no effect, in either strain, on the P. gingivalis growth rate or on the response to hydrogen peroxide or growth at pH 9, at 41 degrees C, or at low hemin availability. Transcriptome analysis using DNA microarrays revealed that inactivation of fimS resulted in the differential expression of 10% of the P. gingivalis genome (>1.5-fold; P fimS mutant. PMID:20061484

Lo, Alvin; Seers, Christine; Dashper, Stuart; Butler, Catherine; Walker, Glenn; Walsh, Katrina; Catmull, Deanne; Hoffmann, Brigitte; Cleal, Steven; Lissel, Patricia; Boyce, John; Reynolds, Eric

2010-03-01

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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

2011-03-01

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Peptidyl Arginine Deiminase from Porphyromonas gingivalis Abolishes Anaphylatoxin C5a Activity.  

Science.gov (United States)

Evasion of killing by the complement system, a crucial part of innate immunity, is a key evolutionary strategy of many human pathogens. A major etiological agent of chronic periodontitis, the Gram-negative bacterium Porphyromonas gingivalis, produces a vast arsenal of virulence factors that compromise human defense mechanisms. One of these is peptidylarginine deiminase (PPAD), an enzyme unique to P. gingivalis among bacteria, which converts Arg residues in polypeptide chains into citrulline. Here, we report that PPAD citrullination of a critical C-terminal arginine of the anaphylatoxin C5a disabled the protein function. Treatment of C5a with PPAD in vitro resulted in decreased chemotaxis of human neutrophils and diminished calcium signaling in monocytic cell line U937 transfected with the C5a receptor (C5aR) and loaded with a fluorescent intracellular calcium probe: Fura-2 AM. Moreover, a low degree of citrullination of internal arginine residues by PPAD was also detected using mass spectrometry. Further, after treatment of C5 with outer membrane vesicles naturally shed by P. gingivalis, we observed generation of C5a totally citrullinated at the C-terminal Arg-74 residue (Arg74Cit). In stark contrast, only native C5a was detected after treatment with PPAD-null outer membrane vesicles. Our study suggests reduced antibacterial and proinflammatory capacity of citrullinated C5a, achieved via lower level of chemotactic potential of the modified molecule, and weaker cell activation. In the context of previous studies, which showed crosstalk between C5aR and Toll-like receptors, as well as enhanced arthritis development in mice infected with PPAD-expressing P. gingivalis, our findings support a crucial role of PPAD in the virulence of P. gingivalis. PMID:25324545

Bielecka, Ewa; Scavenius, Carsten; Kantyka, Tomasz; Jusko, Monika; Mizgalska, Danuta; Szmigielski, Borys; Potempa, Barbara; Enghild, Jan J; Prossnitz, Eric R; Blom, Anna M; Potempa, Jan

2014-11-21

 
 
 
 
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Peptidyl arginine deiminase from Porphyromonas gingivalis abolishes C5a activity  

DEFF Research Database (Denmark)

Evasion of killing by the complement system, a crucial part of innate immunity, is a key evolutionary strategy of many human pathogens. A major etiological agent of chronic periodontitis, the Gram-negative bacterium Porphyromonas gingivalis produces a vast arsenal of virulence factors that compromise human defense mechanisms. One of these is peptidylarginine deiminase (PPAD), an enzyme unique to P. gingivalis among bacteria, which converts Arg residues in polypeptide chains into citrulline. Here, we report that PPAD citrullination of a critical C-terminal arginine of the anaphylatoxin C5a disabled the protein function. Treatment of C5a with PPAD in vitro resulted in decreased chemotaxis of human neutrophils and diminished calcium signaling in monocytic cell line U937 transfected with the C5a receptor (C5aR) and loaded with a fluorescent intracellular calcium probe: Fura 2-AM. Moreover, a low degree of citrullination of internal arginine residues by PPAD was also detected using mass spectrometry. Further, after treatment of C5 with outer membrane vesicles (OMVs) naturally shed by P. gingivalis we observed generation of C5a totally citrullinated at the C-terminal Arg74 residue (Arg74Cit). In stark contrast only native C5a was detected after treatment with PPAD-null OMVs. Our study suggests reduced antibacterial and proinflammatory capacity of citrullinated C5a, achieved via lower level of chemotactic potential of the modified molecule, and weaker cell activation. In the context of previous studies, which showed crosstalk between C5aR and toll-like receptors, as well as enhanced arthritis development in mice infected with PPAD expressing P. gingivalis, our findings support a crucial role of PPAD in the virulence of P. gingivalis.

Bielecka, Ewa; Scavenius, Carsten

2014-01-01

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Porphyromonas gingivalis–dendritic cell interactions: consequences for coronary artery disease  

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Full Text Available An estimated 80 million US adults have one or more types of cardiovascular diseases. Atherosclerosis is the single most important contributor to cardiovascular diseases; however, only 50% of atherosclerosis patients have currently identified risk factors. Chronic periodontitis, a common inflammatory disease, is linked to an increased cardiovascular risk. Dendritic cells (DCs are potent antigen presenting cells that infiltrate arterial walls and may destabilize atherosclerotic plaques in cardiovascular disease. While the source of these DCs in atherosclerotic plaques is presently unclear, we propose that dermal DCs from peripheral inflamed sites such as CP tissues are a potential source. This review will examine the role of the opportunistic oral pathogen Porphyromonas gingivalis in invading DCs and stimulating their mobilization and misdirection through the bloodstream. Based on our published observations, combined with some new data, as well as a focused review of the literature we will propose a model for how P. gingivalis may exploit DCs to gain access to systemic circulation and contribute to coronary artery disease. Our published evidence supports a significant role for P. gingivalis in subverting normal DC function, promoting a semimature, highly migratory, and immunosuppressive DC phenotype that contributes to the inflammatory development of atherosclerosis and, eventually, plaque rupture.

Amir E. Zeituni

2010-12-01

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Porphyromonas gingivalis peptidoglycans induce excessive activation of the innate immune system in silkworm larvae.  

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Porphyromonas gingivalis, a pathogen that causes inflammation in human periodontal tissue, killed silkworm (Bombyx mori, Lepidoptera) larvae when injected into the blood (hemolymph). Silkworm lethality was not rescued by antibiotic treatment, and heat-killed bacteria were also lethal. Heat-killed bacteria of mutant P. gingivalis strains lacking virulence factors also killed silkworms. Silkworms died after injection of peptidoglycans purified from P. gingivalis (pPG), and pPG toxicity was blocked by treatment with mutanolysin, a peptidoglycan-degrading enzyme. pPG induced silkworm hemolymph melanization at the same dose as that required to kill the animal. pPG injection increased caspase activity in silkworm tissues. pPG-induced silkworm death was delayed by injecting melanization-inhibiting reagents (a serine protease inhibitor and 1-phenyl-2-thiourea), antioxidants (N-acetyl-l-cysteine, glutathione, and catalase), and a caspase inhibitor (Ac-DEVD-CHO). Thus, pPG induces excessive activation of the innate immune response, which leads to the generation of reactive oxygen species and apoptotic cell death in the host tissue. PMID:20702417

Ishii, Kenichi; Hamamoto, Hiroshi; Imamura, Katsutoshi; Adachi, Tatsuo; Shoji, Mikio; Nakayama, Koji; Sekimizu, Kazuhisa

2010-10-22

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

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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. PMID:23352947

Zhang, Dan; Ren, Ling; Zhang, Yang; Xue, Nan; Yang, Ke; Zhong, Ming

2013-05-01

65

Structure determination and analysis of a haemolytic gingipain adhesin domain from Porphyromonas gingivalis  

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Porphyromonas gingivalis is an obligately anaerobic bacterium recognized as an aetiological agent of adult periodontitis. P. gingivalis produces cysteine proteinases, the gingipains. The crystal structure of a domain within the haemagglutinin region of the lysine gingipain (Kgp) is reported here. The domain was named K2 as it is the second of three homologous structural modules in Kgp. The K2 domain structure is a 'jelly-roll' fold with two anti-parallel {beta}-sheets. This fold topology is shared with adhesive domains from functionally diverse receptors such as MAM domains, ephrin receptor ligand binding domains and a number of carbohydrate binding modules. Possible functions of K2 were investigated. K2 induced haemolysis of erythrocytes in a dose-dependent manner that was augmented by the blocking of anion transport. Further, cysteine-activated arginine gingipain RgpB, which degrades glycophorin A, sensitized erythrocytes to the haemolytic effect of K2. Cleaved K2, similar to that found in extracted Kgp, lacks the haemolytic activity indicating that autolysis of Kgp may be a staged process which is artificially enhanced by extraction of the protein. The data indicate a functional role for K2 in the integrated capacity conferred by Kgp to enable the porphyrin auxotroph P. gingivalis to capture essential haem from erythrocytes.

Li, N.; Yun, P.; Nadkarni, M.A.; Ghadikolaee, N.B.; Nguyen, K.A.; Lee, M.; Hunter, N.; Collyer, C.A. (Sydney)

2010-08-27

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Periodontitis, Porphyromonas gingivalis y su relación con la expresión de quorum sensing / Periodontitis, Porphyromonas gingivalis and its relation to quorum sensing expression  

Scientific Electronic Library Online (English)

Full Text Available SciELO Cuba | Language: Spanish Abstract in spanish Los mecanismos de señalización bacteriana desempeñan un papel fundamental en el establecimiento y progresión de la enfermedad periodontal. Dadas estas circunstancias es crucial profundizar en el entendimiento de estos mecanismos para intentar proveer estrategias terapéuticas novedosas. El presente a [...] rtículo de revisión, de carácter narrativo, tiene como objetivo conducir un análisis crítico de la evidencia disponible sobre la influencia de Porphyromonas gingivalis (Pg) y expresión de quorum sensing (Qs) en enfermedad periodontal. Se realizó una búsqueda a través de bases de datos como Ovid (MEDLINE), ScienceDirect, Hinari. El conocimiento actual de estos mecanismos ofrece la posibilidad de desarrollar nuevos y profundos estudios (teóricos y experimentales) sobre la expresión del QS en pacientes con enfermedad periodontal y permitirá un novedoso campo de investigación con el que no se cuenta en la actualidad. Desde su descubrimiento, el QS se vislumbra como un espacio de investigación valioso en el cual se debe insistir de manera permanente. La anterior evidencia permite concluir que a través de la regulación de la expresión de determinados genes en bacterias como la PG, se puede efectuar la inhibición de la formación de las biopelículas que tiene efectos directos e indirectos sobre el desarrollo de la enfermedad periodontal. Abstract in english The bacterial signaling mechanisms play a key role in the establishment and progression of periodontal disease. Due to these circumstances it is crucial to deepen in the understanding of these mechanisms to try to provide novel therapeutic strategies. The objective of present narrative literature re [...] view was to make a critical analyze of the available evidence on the influence of Porphyromonas gingivalis (PG) and the quorum sensing expression in periodontal disease. Using the Ovid (MEDLINE) ScienceDirect, Hinari database we made a search. The current knowledge of these mechanisms offers the possibility of developing new and deep studies (theoretical and experimental) on the QS expression in patients presenting with periodontal disease allowing a novel research field not currently available. From its discovery the QS is discerned as a valuable research space in which we must to insist in a permanent way. The above mentioned evidence allows concluding that by the regulation of the expression of determined genes in bacteria like PG, it is possible to carry out the inhibition in the formation of the biofilms with direct and indirect effects on the periodontal disease development.

Antonio, Díaz Caballero; Ricardo, Vivas Reyes; Leonardo, Puerta Llerena; Maicol, Ahumedo Monterrosa; Ricardo, Cabrales Salgado; Alejandra, Herrera Herrera; Miguel, Simancas Pallares.

2010-12-01

67

The relationship between colonization and haemagglutination inhibiting and B cell epitopes of Porphyromonas gingivalis  

Science.gov (United States)

Passive immunization with the monoclonal antibody 61BG1.3 selectively prevents colonization by Porphyromonas gingivalis in humans (Booth V, Ashley FP, Lehner T. Infect Immun 1996; 64:422-7). The protective MoAb recognizes the j3 component of the RI protease of P. gingivalis which is formed by proteolytic processing of a polyprotein precursor termed PrpRl. This subunit is both a haemagglutinin and an antigen which is recognized by sera from patients with periodontitis. In this study the relationship was investigated between a colonization epitope which is recognized by the MoAb 61BG1.3, a haemagglutinating and B cell epitope which are recognized by sera from patients with periodontitis. B cell epitopes were mapped by Western blotting with a series of truncated recombinant polypeptides spanning the adhesion domain within residues 784–1130 of PrpRl and by ELISA using a panel of synthetic peptides spanning the same sequence. The epitope which is recognized by the protective MoAb was mapped within residues 907–931 of PrpRl, while serum responses of patients were directed predominantly to the adjacent carboxy-terminal sequence within residues 934–1042. The haemagglutinating epitope was mapped to residues 1073–1112. In view of our previous findings that the MoAb 61BG1.3 prevents colonization of P. gingivalis in vivo and inhibits haemagglutination, these two epitopes may be in proximity in the native protein. Active or passive immunization strategies which target the protective or haemagglutinating epitopes of the adhesion domain of PrpRl may provide a means of preventing infection with P. gingivalis. PMID:9367414

KELLY, C G; BOOTH, V; KENDAL, H; SLANEY, J M; CURTIS, M A; LEHNER, T

1997-01-01

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Identification and characterization of prokaryotic dipeptidyl-peptidase 5 from Porphyromonas gingivalis.  

Science.gov (United States)

Porphyromonas gingivalis, a Gram-negative asaccharolytic anaerobe, is a major causative organism of chronic periodontitis. Because the bacterium utilizes amino acids as energy and carbon sources and incorporates them mainly as dipeptides, a wide variety of dipeptide production processes mediated by dipeptidyl-peptidases (DPPs) should be beneficial for the organism. In the present study, we identified the fourth P. gingivalis enzyme, DPP5. In a dpp4-7-11-disrupted P. gingivalis ATCC 33277, a DPP7-like activity still remained. PGN_0756 possessed an activity indistinguishable from that of the mutant, and was identified as a bacterial orthologue of fungal DPP5, because of its substrate specificity and 28.5% amino acid sequence identity with an Aspergillus fumigatus entity. P. gingivalis DPP5 was composed of 684 amino acids with a molecular mass of 77,453, and existed as a dimer while migrating at 66 kDa on SDS-PAGE. It preferred Ala and hydrophobic residues, had no activity toward Pro at the P1 position, and no preference for hydrophobic P2 residues, showed an optimal pH of 6.7 in the presence of NaCl, demonstrated Km and kcat/Km values for Lys-Ala-MCA of 688 ?M and 11.02 ?M(-1) s(-1), respectively, and was localized in the periplasm. DPP5 elaborately complemented DPP7 in liberation of dipeptides with hydrophobic P1 residues. Examinations of DPP- and gingipain gene-disrupted mutants indicated that DPP4, DPP5, DPP7, and DPP11 together with Arg- and Lys-gingipains cooperatively liberate most dipeptides from nutrient oligopeptides. This is the first study to report that DPP5 is expressed not only in eukaryotes, but also widely distributed in bacteria and archaea. PMID:24398682

Ohara-Nemoto, Yuko; Rouf, Shakh M A; Naito, Mariko; Yanase, Amie; Tetsuo, Fumi; Ono, Toshio; Kobayakawa, Takeshi; Shimoyama, Yu; Kimura, Shigenobu; Nakayama, Koji; Saiki, Keitarou; Konishi, Kiyoshi; Nemoto, Takayuki K

2014-02-28

69

Human lactoferrin binding to Porphyromonas gingivalis, Prevotella intermedia and Prevotella melaninogenica.  

Science.gov (United States)

Human isolates of Porphyromonas gingivalis (n = 16), Prevotella intermedia n = 82) and Prevotella melaninogenica (n = 18) from diseased periodontal pockets were examined for interaction with human lactoferrin (HLf) in a standardized 125I-labeled protein binding assay. The highest HLf binding was found in P. intermedia strains, followed by P. gingivalis and P. melaninogenica. Further characterization of the interaction was performed with 1 representative strain from each species. HLf binding to P. gingivalis reached a saturation instantly and was optimal at pH 5.0-6.5. The corresponding values for P. melaninogenica were 90 min and pH 3.0-5.5. The HLf binding to the 2 strains seem to be nonspecific. In contrast, P. intermedia demonstrated specific binding, and a time-saturability within 60 min with an optimal uptake at pH 6.0-7.5. Scatchard analysis implied 45,000 receptors per cell with an affinity constant of 5.5 x 10(-7) M on P. intermedia strain 4H. The binding capacity in all 3 strains was affected by the culture medium. HLf binding components in these strains were susceptible to heat or proteases. Binding was eliminated in P. gingivalis and was enhanced in P. intermedia and P. melaninogenica by periodate treatment. Unlabeled HLf or bovine lactoferrin effectively displaced labeled HLf binding. Various proteins and carbohydrates did not inhibit HLf binding. Our data suggest that HLf binds to these periodontitis-associated species and that this mechanism is distinct from the previously known ligand interactions in oral bacteria. PMID:1726544

Kalfas, S; Andersson, M; Edwardsson, S; Forsgren, A; Naidu, A S

1991-12-01

70

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

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Full Text Available 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 extract e descritores clínicos periodontais. Material e Métodos: Indivíduos com periodontite (29 e com saúde periodontal (26 foram avaliados de acordo com as medidas de profundidade de sondagem, sangramento à sondagem e nível de inserção clínica. O extrato de Porphyromonas gingivalis foi fracionado por cromatografia de troca iônica e a resposta humoral contra a fração IV foi avaliada por “enzyme linked immunosorbent assay”. Resultados e Conclusão: O percentual de sítios com sangramento à sondagem (critério 1 estava correlacionado significativamente com os níveis séricos de IgG2 (r = 0,385; p < 0,05. Os níveis de IgG total e IgG2 correlacionaram-se significativamente com o percentual de sítios com nível de inserção clínica (NIC ? 3 mm (critério 2 (r = 0,428; p < 0,05 e r = 0,510; p < 0,01, respectivamente, NIC ? 5 mm (critérion 3 (r = 0,499 e r = 0,518, respectivamente; p < 0,01 e percentual de sítios com NIC ? 3 mm associado a profundidade de sondagem ? 4 mm e sangramento à sondagem no mesmo sítio (criterion 4 (r = 0,607; p < 0,001 e r = 0,487; p < 0,01, respectivamente. Foi observada uma correlação positiva estatistivamente significante entre os níveis de IgGA e IgG1 e o critério 4 (r = 0,339 e r = 0,345, respectivamente; p < 0,05, e entre os níveis de IgG3 e o critério 3 (p < 0,05; r = 0,370. Estes resultados indicam que quanto mais acurado for o critério de diagnóstico empregado para a determinação da doença periodontal, maiores os níveis séricos de imunoglobulinas.

Trindade, Soraya Castro et. al

2007-01-01

71

Sulfonamide inhibition studies of the ?-carbonic anhydrase from the oral pathogen Porphyromonas gingivalis.  

Science.gov (United States)

A carbonic anhydrase (CA, EC 4.2.1.1) denominated PgiCA, belonging to the ?-class, from the oral pathogenic bacteria Porphyromonas gingivalis, the main causative agent of periodontitis, was investigated for its inhibition profile with sulfonamides and one sulfamate. Dichlorophenamide, topiramate and many simple aromatic/heterocyclic sulfonamides were ineffective as PgiCA inhibitors whereas the best inhibition was observed with halogenosulfanilamides incorporating heavy halogens, 4-hydroxy- and 4-hydroxyalkyl-benzenesulfonamides, acetazolamide, methazolamide, zonisamide, indisulam, celecoxib, saccharin and hydrochlorothiazide (KIs in the range of 131-380nM). The inhibition profile of PgiCA was very different from that of CAM, hCA I and II or the ?-CA from a protozoan parasite (Leishmania donovani chagasii). Identification of potent and possibly selective inhibitors of PgiCA may lead to pharmacological tools useful for understanding the physiological role(s) of this enzyme. PMID:24316122

Vullo, Daniela; Del Prete, Sonia; Osman, Sameh M; De Luca, Viviana; Scozzafava, Andrea; Alothman, Zeid; Supuran, Claudiu T; Capasso, Clemente

2014-01-01

72

Diagnostic evaluation of a nanobody with picomolar affinity toward the protease RgpB from Porphyromonas gingivalis  

DEFF Research Database (Denmark)

Porphyromonas gingivalis is one of the major periodontitis-causing pathogens. P. gingivalis secretes a group of proteases termed gingipains, and in this study we have used the RgpB gingipain as a biomarker for P. gingivalis. We constructed a naive camel nanobody library and used phage display to select one nanobody toward RgpB with picomolar affinity. The nanobody was used in an inhibition assay for detection of RgpB in buffer as well as in saliva. The nanobody was highly specific for RgpB given that it did not bind to the homologous gingipain HRgpA. This indicated the presence of a binding epitope within the immunoglobulin-like domain of RgpB. A subtractive inhibition assay was used to demonstrate that the nanobody could bind native RgpB in the context of intact cells. The nanobody bound exclusively to the P. gingivalis membrane-bound RgpB isoform (mt-RgpB) and to secreted soluble RgpB. Further cross-reactivity studies with P. gingivalis gingipain deletion mutants showed that the nanobody could discriminate between native RgpB and native Kgp and RgpA in complex bacterial samples. This study demonstrates that RgpB can be used as a specific biomarker for P. gingivalis detection and that the presented nanobody-based assay could supplement existing methods for P. gingivalis detection.

Skottrup, Peter Durand; Leonard, Paul

2011-01-01

73

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

Science.gov (United States)

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. PMID:23264054

Maeda, Kazuhiko; Nagata, Hideki; Kuboniwa, Masae; Ojima, Miki; Osaki, Tsukasa; Minamino, Naoto; Amano, Atsuo

2013-03-01

74

Altered Gingipain Maturation in vimA- and vimE-Defective Isogenic Mutants of Porphyromonas gingivalis  

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We have previously shown that gingipain activity in Porphyromonas gingivalis is modulated by the unique vimA and vimE genes. To determine if these genes had a similar phenotypic effect on protease maturation and activation, isogenic mutants defective in those genes were further characterized. Western blot analyses with antigingipain antibodies showed RgpA-, RgpB-, and Kgp-immunoreactive bands in membrane fractions as well as the culture supernatant of both P. gingivalis W83 and FLL93, the vim...

Vanterpool, Elaine; Roy, Francis; Sandberg, Lawrence; Fletcher, Hansel M.

2005-01-01

75

Genetic exchange of fimbrial alleles exemplifies the adaptive virulence strategy of Porphyromonas gingivalis.  

Science.gov (United States)

Porphyromonas gingivalis is a gram-negative anaerobic bacterium, a member of the human oral microbiome, and a proposed "keystone" pathogen in the development of chronic periodontitis, an inflammatory disease of the gingiva. P. gingivalis is a genetically diverse species, and is able to exchange chromosomal DNA between strains by natural competence and conjugation. In this study, we investigate the role of horizontal DNA transfer as an adaptive process to modify behavior, using the major fimbriae as our model system, due to their critical role in mediating interactions with the host environment. We show that P. gingivalis is able to exchange fimbrial allele types I and IV into four distinct strain backgrounds via natural competence. In all recombinants, we detected a complete exchange of the entire fimA allele, and the rate of exchange varies between the different strain backgrounds. In addition, gene exchange within other regions of the fimbrial genetic locus was identified. To measure the biological implications of these allele swaps we compared three genotypes of fimA in an isogenic background, strain ATCC 33277. We demonstrate that exchange of fimbrial allele type results in profound phenotypic changes, including the quantity of fimbriae elaborated, membrane blebbing, auto-aggregation and other virulence-associated phenotypes. Replacement of the type I allele with either the type III or IV allele resulted in increased invasion of gingival fibroblast cells relative to the isogenic parent strain. While genetic variability is known to impact host-microbiome interactions, this is the first study to quantitatively assess the adaptive effect of exchanging genes within the pan genome cloud. This is significant as it presents a potential mechanism by which opportunistic pathogens may acquire the traits necessary to modify host-microbial interactions. PMID:24626479

Kerr, Jennifer E; Abramian, Jared R; Dao, Doan-Hieu V; Rigney, Todd W; Fritz, Jamie; Pham, Tan; Gay, Isabel; Parthasarathy, Kavitha; Wang, Bing-yan; Zhang, Wenjian; Tribble, Gena D

2014-01-01

76

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

Energy Technology Data Exchange (ETDEWEB)

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

77

Inactivation of epidermal growth factor by Porphyromonas gingivalis as a potential mechanism for periodontal tissue damage  

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Porphyromonas gingivalis is a Gram-negative bacterium associated with the development of periodontitis. The evolutionary success of this pathogen results directly from the presence of numerous virulence factors, including a peptidylarginine deiminase (PPAD), an enzyme, which converts arginine to citrulline in proteins and peptides. Such posttranslational modification is thought to affect the function of many different signaling molecules. Taking into account the importance of tissue remodeling and repair mechanisms for periodontal homeostasis, which are orchestrated by ligands of the epidermal growth factor receptor (EGFR), we investigated the ability of PPAD to distort cross-talk between the epithelium and the EGF signaling pathway. We found that EGF preincubation with purified recombinant PPAD, or a wild-type strain of P. gingivalis, but not with a PPAD-deficient isogenic-mutant, efficiently hindered the ability of the growth factor to stimulate epidermal cell proliferation and migration. In addition, PPAD abrogated EGFR-EGF interaction-dependent stimulation of expression of Suppressor of Cytokine Signaling 3 (SOCS3) and Interferon Regulatory Factor 1 (IRF-1). Biochemical analysis clearly showed that the PPAD-exerted effects on EGF activities were solely due to deimination of the C-terminal arginine. Interestingly, citrullination of two internal Arg residues with human endogenous peptidylarginine deiminases did not alter EFG function, arguing that the C-terminal arginine is essential for EGF biological activity. Cumulatively, these data suggest that PPAD-activity-abrogating EGF function in gingival pockets may at least partially contribute to tissue damage and delayed healing within P. gingivalis-infected periodontia.

Pyrc, Krzysztof; Milewska, Aleksandra

2013-01-01

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Insights into the antiatherogenic molecular mechanisms of andrographolide against Porphyromonas gingivalis-induced atherosclerosis in rabbits.  

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Atherosclerosis is the commonest and most important vascular disease. Andrographolide (AND) is the main bioactive component of the medicinal plant Andrographis paniculata and is used in traditional medicine. This study was aimed to evaluate the antiatherogenic effect of AND against atherosclerosis induced by Porphyromonas gingivalis in White New Zealand rabbits. Thirty rabbits were divided into five groups as follows: G1, normal group; G2-5, were orally challenged with P. gingivalis five times a week over 12 weeks; G2, atherogenic control group; G3, standard group treated with atorvastatin (AV) 5 mg/kg; and G4 and G5, treatment groups treated with AND 10 and 20 mg/kg, respectively over 12 weeks. Serums were subjected to antioxidant enzymatic and anti-inflammatory activities, and the aorta was subjected to histological analyses. Groups treated with AND showed a significant reversal of liver and renal biochemical changes, compared with the atherogenic control group. In the same groups, superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), total glutathione (GSH) levels in serum were significantly increased (P?gingivalis. PMID:25172523

Al Batran, Rami; Al-Bayaty, Fouad; Al-Obaidi, Mazen M Jamil; Ashrafi, Amer

2014-12-01

79

Tetratricopeptide repeat protein-associated proteins contribute to the virulence of Porphyromonas gingivalis.  

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Porphyromonas gingivalis is one of the most etiologically important microorganisms in periodontal disease. We found in a previous study that PG1385 (TprA) protein, a tetratricopeptide repeat (TPR) protein, was upregulated in P. gingivalis wild-type cells placed in a mouse subcutaneous chamber and that a tprA mutant was clearly less virulent in the mouse subcutaneous abscess model (M. Yoshimura et al., Oral Microbiol. Immunol. 23:413-418, 2008). In the present study, we investigated the gene expression profile of tprA mutant cells placed in a mouse subcutaneous chamber and found that 9 genes, including PG2102 (tapA), PG2101 (tapB), and PG2100 (tapC) genes, were downregulated in the tprA mutant compared with those in the wild type. Expression of a cluster of tapA, tapB, and tapC genes of the mutant was also downregulated in an in vitro culture with enriched brain heart infusion medium. The TprA protein has three TPR motifs known as a protein-protein interaction module. Yeast two-hybrid system analysis and in vitro protein binding assays with immunoprecipitation and surface plasmon resonance detection revealed that the TprA protein could bind to TapA and TapB proteins. TprA and TapB proteins were located in the periplasmic space, whereas TapA, which appeared to be one of the C-terminal domain family proteins, was located at the outer membrane. We constructed tapA, tapB, and tapC single mutants and a tapA-tapB-tapC deletion mutant. In the mouse subcutaneous infection experiment, all of the mutants were less virulent than the wild type. These results suggest that TprA, TapA, TapB, and TapC are cooperatively involved in P. gingivalis virulence. PMID:20351137

Kondo, Yoshio; Ohara, Naoya; Sato, Keiko; Yoshimura, Mamiko; Yukitake, Hideharu; Naito, Mariko; Fujiwara, Taku; Nakayama, Koji

2010-06-01

80

Immune response of macrophages from young and aged mice to the oral pathogenic bacterium Porphyromonas gingivalis  

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Abstract Periodontal disease is a chronic inflammatory gum disease that in severe cases leads to tooth loss. Porphyromonas gingivalis (Pg) is a bacterium closely associated with generalized forms of periodontal disease. Clinical onset of generalized periodontal disease commonly presents in individuals over the age of 40. Little is known regarding the effect of aging on inflammation associated with periodontal disease. In the present study we examined the immune response of ...

Gibson Frank C; Kantarci Alpdogan; Shaik-Dasthagirisaheb Yazdani B

2010-01-01

 
 
 
 
81

Inhibition of gingipains by their profragments as the mechanism protecting Porphyromonas gingivalis against premature activation of secreted proteases  

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Arginine-specific (RgpB and RgpA) and lysine-specific (Kgp) gingipains are secretory cysteine proteinases of Porphyromonas gingivalis that act as important virulence factors for the organism. They are translated as zymogens with both N- and C-terminal extensions, which are proteolytically cleaved during secretion. In this report, we describe and characterize inhibition of the gingipains by their N-terminal prodomains to maintain latency during their export through the cellular compartments.

Veillard, Florian; Sztukowska, Maryta

2013-01-01

82

Effect of Porphyromonas gingivalis infection on post-transcriptional regulation of the low-density lipoprotein receptor in mice  

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Full Text Available Abstract Background Periodontal disease is suggested to increase the risk of atherothrombotic disease by inducing dyslipidemia. Recently, we demonstrated that proprotein convertase subtilisin/kexin type 9 (PCSK9, which is known to play a critical role in the regulation of circulating low-density lipoprotein (LDL cholesterol levels, is elevated in periodontitis patients. However, the underlying mechanisms of elevation of PCSK9 in periodontitis patients are largely unknown. Here, we explored whether Porphyromonas gingivalis, a representative periodontopathic bacterium, -induced inflammatory response regulates serum PCSK9 and cholesterol levels using animal models. Methods We infected C57BL/6 mice intraperitoneally with Porphyromonas gingivalis, a representative strain of periodontopathic bacteria, and evaluated serum PCSK9 levels and the serum lipid profile. PCSK9 and LDL receptor (LDLR gene and protein expression, as well as liver X receptors (Lxrs, inducible degrader of the LDLR (Idol, and sterol regulatory element binding transcription factor (Srebf2 gene expression, were examined in the liver. Results P. gingivalis infection induced a significant elevation of serum PCSK9 levels and a concomitant elevation of total and LDL cholesterol compared with sham-infected mice. The LDL cholesterol levels were significantly correlated with PCSK9 levels. Expression of the Pcsk9, Ldlr, and Srebf2 genes was upregulated in the livers of the P. gingivalis-infected mice compared with the sham-infected mice. Although Pcsk9 gene expression is known to be positively regulated by sterol regulatory element binding protein (SREBP2 (human homologue of Srebf2, whereas Srebf2 is negatively regulated by cholesterol, the elevated expression of Srebf2 found in the infected mice is thought to be mediated by P. gingivalis infection. Conclusions P. gingivalis infection upregulates PCSK9 production via upregulation of Srebf2, independent of cholesterol levels. Further studies are required to elucidate how infection regulates Srebf2 expression and subsequently influences lipid metabolism.

Miyazawa Haruna

2012-09-01

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Porphyromonas gingivalis Peptidylarginine Deiminase, a Key Contributor in the Pathogenesis of Experimental Periodontal Disease and Experimental Arthritis  

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Objectives To investigate the suggested role of Porphyromonas gingivalis peptidylarginine deiminase (PAD) in the relationship between the aetiology of periodontal disease and experimentally induced arthritis and the possible association between these two conditions. Methods A genetically modified PAD-deficient strain of P. gingivalis W50 was produced. The effect of this strain, compared to the wild type, in an established murine model for experimental periodontitis and experimental arthritis was assessed. Experimental periodontitis was induced following oral inoculation with the PAD-deficient and wild type strains of P. gingivalis. Experimental arthritis was induced via the collagen antibody induction process and was monitored by assessment of paw swelling and micro-CT analysis of the radio-carpal joints. Experimental periodontitis was monitored by micro CT scans of the mandible and histological assessment of the periodontal tissues around the mandibular molars. Serum levels of anti-citrullinated protein antibodies (ACPA) and P. gingivalis were assessed by ELISA. Results The development of experimental periodontitis was significantly reduced in the presence of the PAD-deficient P. gingivalis strain. When experimental arthritis was induced in the presence of the PAD-deficient strain there was less paw swelling, less erosive bone damage to the joints and reduced serum ACPA levels when compared to the wild type P. gingivalis inoculated group. Conclusion This study has demonstrated that a PAD-deficient strain of P. gingivalis was associated with significantly reduced periodontal inflammation. In addition the extent of experimental arthritis was significantly reduced in animals exposed to prior induction of periodontal disease through oral inoculation of the PAD-deficient strain versus the wild type. This adds further evidence to the potential role for P. gingivalis and its PAD in the pathogenesis of periodontitis and exacerbation of arthritis. Further studies are now needed to elucidate the mechanisms which drive these processes. PMID:24959715

Gully, Neville; Bright, Richard; Marino, Victor; Marchant, Ceilidh; Cantley, Melissa; Haynes, David; Butler, Catherine; Dashper, Stuart; Reynolds, Eric; Bartold, Mark

2014-01-01

84

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

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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. PMID:23995804

Nagano, Keiji

2013-01-01

85

Porphyromonas gingivalis GroEL Induces Osteoclastogenesis of Periodontal Ligament Cells and Enhances Alveolar Bone Resorption in Rats  

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Porphyromonas gingivalis is a major periodontal pathogen that contains a variety of virulence factors. The antibody titer to P. gingivalis GroEL, a homologue of HSP60, is significantly higher in periodontitis patients than in healthy control subjects, suggesting that P. gingivalis GroEL is a potential stimulator of periodontal disease. However, the specific role of GroEL in periodontal disease remains unclear. Here, we investigated the effect of P. gingivalis GroEL on human periodontal ligament (PDL) cells in vitro, as well as its effect on alveolar bone resorption in rats in vivo. First, we found that stimulation of PDL cells with recombinant GroEL increased the secretion of the bone resorption-associated cytokines interleukin (IL)-6 and IL-8, potentially via NF-?B activation. Furthermore, GroEL could effectively stimulate PDL cell migration, possibly through activation of integrin ?1 and ?2 mRNA expression as well as cytoskeletal reorganization. Additionally, GroEL may be involved in osteoclastogenesis via receptor activator of nuclear factor ?-B ligand (RANKL) activation and alkaline phosphatase (ALP) mRNA inhibition in PDL cells. Finally, we inoculated GroEL into rat gingiva, and the results of microcomputed tomography (micro-CT) and histomorphometric assays indicated that the administration of GroEL significantly increased inflammation and bone loss. In conclusion, P. gingivalis GroEL may act as a potent virulence factor, contributing to osteoclastogenesis of PDL cells and resulting in periodontal disease with alveolar bone resorption. PMID:25058444

Lin, Feng-Yen; Hsiao, Fung-Ping; Huang, Chun-Yao; Shih, Chun-Ming; Tsao, Nai-Wen; Tsai, Chien-Sung; Yang, Shue-Fen; Chang, Nen-Chung; Hung, Shan-Ling; Lin, Yi-Wen

2014-01-01

86

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

2010-04-01

87

Evaluation of the effect of andrographolide on atherosclerotic rabbits induced by Porphyromonas gingivalis.  

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Epidemiologic evidence has demonstrated significant associations between atherosclerosis and Porphyromonas gingivalis (Pg). We had investigated the effect of andrographolide (AND) on atherosclerosis induced by Pg in rabbits. For experimental purpose, we separated thirty male white New Zealand rabbits into 5 groups. Group 1 received standard food pellets; Groups 2-5 were orally challenged with Pg; Group 3 received atorvastatin (AV, 5?mg/kg), and Groups 4-5 received 10 and 20?mg/kg of AND, respectively, over 12 weeks. Groups treated with AND showed significant decrease in TC, TG, and LDL levels (P<0.05) and significant increase in HDL level in the serum of rabbits. Furthermore, the treated groups (G3-G5) exhibited reductions in interleukins (IL-1? and IL-6) and C-reactive protein (CRP) as compared to atherogenicgroup (G2). The histological results showed that the thickening of atherosclerotic plaques were less significant in treated groups (G3-G5) compared with atherogenicgroup (G2). Also, alpha-smooth muscle actin (?-SMA) staining decreased within the plaques of atherogenicgroup (G2), while it was increased in treated groups (G3-G5). Lastly, groups treated with AV and AND (G3-G5) showed significant reduction of CD36 expression (P<0.05) compared to atherogenicgroup (G2). These results substantially proved that AND contain antiatherogenic activity. PMID:25215291

Al Batran, Rami; Al-Bayaty, Fouad; Al-Obaidi, Mazen M Jamil; Hussain, Saba F; Mulok, Tengku Z

2014-01-01

88

Effects of a combination therapy to eliminate Porphyromonas gingivalis in refractory periodontitis.  

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This report describes the clinical and microbiological features of 30 refractory patients and their response to a combined local and systemic therapy at 6 weeks and 3 years following treatment. The refractory treatment protocol (RefTx) consisted of a 2-week regimen of amoxicillin/clavulanate potassium in conjunction with professional, intrasulcular delivery of povidone iodine, and chlorhexide mouthwash rinses b.i.d. Eighty-seven percent of the patients had favorable clinical responses to the RefTx and could be divided into 3 groups (A, B, C) based upon initial flora patterns and the shifts that occurred following treatment. Pretreatment prevalence of Porphyromonas gingivalis (P.g.) was 36.7%. The RefTx was effective in reducing P.g. below detection levels in 10 of the 11 positive patients at P effective at reducing probing pocket depth with a 56% decrease in the number of pockets greater than 6 mm at 6 weeks. This was accompanied by an overall gain of > or = 1 mm of probable attachment in 45% of all sites. The clinical effects of the RefTx were shown to persist at 34.3 months with an apparent attachment gain of > or = 1 mm in 41.2% of sites. These data suggest that P.g. and OBP are important pathogens in refractory periodontitis and that the RefTx protocol is an acceptable, non-invasive alternative for the management of these patients. PMID:8277411

Collins, J G; Offenbacher, S; Arnold, R R

1993-10-01

89

Rapid detection of Actinobacillus actinomycetemcomitans, Prevotella intermedia and Porphyromona gingivalis by multiplex PCR.  

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The identification of specific periodontal pathogens by conventional methods, mainly anaerobic cultivation, is difficult, time consuming and even sometimes unreliable. Therefore, a multiplex PCR method for simultaneous detection of Actinobacillus actinomycetemcomitans (A.a.), Porphyromona gingivalis (P.g.) and Prevotella intermedia (P.i.) was developed for rapid and easy identification of these specific bacterial pathogens in subgingival plaque samples. In this paper, there is a detailed description of the oligonucleotide primer selection, DNA extraction and PCR conditions and the sequencing of the amplified products. The locus chosen to be amplified is a highly variable region in the 16S ribosomal DNA. For the development of this technique ATCC cultures and pure cultures from subgingival plaque samples taken from periodontitis patients were used. As an internal positive control a recombinant plasmid was developed. This simple DNA extraction procedure and the DNA amplification and visualization of the amplified product permits the detection of the bacteria in a working day. Thus, this multiplex PCR method is a rapid and effective detection method for specific periodontal pathogens. PMID:9524322

García, L; Tercero, J C; Legido, B; Ramos, J A; Alemany, J; Sanz, M

1998-01-01

90

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

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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

2012-02-01

91

Binding Specificity of the Porphyromonas gingivalis Heme and Hemoglobin Receptor HmuR, Gingipain K, and Gingipain R1 for Heme, Porphyrins, and Metalloporphyrins  

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Previous genetic and biochemical studies have confirmed that hemoglobin and hemin utilization in Porphyromonas gingivalis is mediated by the outer membrane hemoglobin and heme receptor HmuR, as well as gingipain K (Kgp), a lysine-specific cysteine protease, and gingipain R1 (HRgpA), one of two arginine-specific cysteine proteases. In this study we report on the binding specificity of the recombinant P. gingivalis HmuR protein and native gingipains for hemoglobin, hemin, various porphyrins, an...

Olczak, Teresa; Dixon, Dabney White; Genco, Caroline Attardo

2001-01-01

92

Binding of Porphyromonas gingivalis Fimbriae to Proline-Rich Glycoproteins in Parotid Saliva via a Domain Shared by Major Salivary Components  

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Porphyromonas gingivalis, a putative periodontopathogen, can bind to human saliva through its fimbriae. We previously found that salivary components from the submandibular and sublingual glands bind to P. gingivalis fimbriae and that acidic proline-rich protein (PRP) and statherin function as receptor molecules for fimbriae. In this study, we investigated the fimbria-binding components in parotid saliva. Fractionated human parotid saliva by gel-filtration chromatography was immobilized onto n...

Amano, Atsuo; Shizukuishi, Satoshi; Horie, Hiroshi; Kimura, Shigenobu; Morisaki, Ichijiro; Hamada, Shigeyuki

1998-01-01

93

Inactivation of vimF, a Putative Glycosyltransferase Gene Downstream of vimE, Alters Glycosylation and Activation of the Gingipains in Porphyromonas gingivalis W83  

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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-sp...

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

2005-01-01

94

Porphyromonas gingivalis short fimbriae are regulated by a FimS/FimR two-component system.  

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Porphyromonas gingivalis possesses two distinct fimbriae. The long (FimA) fimbriae have been extensively studied. Expression of the fimA gene is tightly controlled by a two-component system (FimS/FimR) through a cascade regulation. The short (Mfa1) fimbriae are less understood. The authors have recently demonstrated that both fimbriae are required for formation of P. gingivalis biofilms. Here, the novel finding that FimR, a member of the two-component regulatory system, is a transcriptional activator of the mfa1 gene is promoted. Unlike the regulatory mechanism of FimA by FimR, this regulation of the mfa1 gene is accomplished by FimR directly binding to the promoter region of mfa1. PMID:17451448

Wu, Jie; Lin, Xinghua; Xie, Hua

2007-06-01

95

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.

Dietrich Elizabeth A

2008-03-01

96

The periodontal pathogen Porphyromonas gingivalis induces expression of transposases and cell death of Streptococcus mitis in a biofilm model.  

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Oral microbial communities are extremely complex biofilms with high numbers of bacterial species interacting with each other (and the host) to maintain homeostasis of the system. Disturbance in the oral microbiome homeostasis can lead to either caries or periodontitis, two of the most common human diseases. Periodontitis is a polymicrobial disease caused by the coordinated action of a complex microbial community, which results in inflammation of tissues that support the teeth. It is the most common cause of tooth loss among adults in the United States, and recent studies have suggested that it may increase the risk for systemic conditions such as cardiovascular diseases. In a recent series of papers, Hajishengallis and coworkers proposed the idea of the "keystone-pathogen" where low-abundance microbial pathogens (Porphyromonas gingivalis) can orchestrate inflammatory disease by turning a benign microbial community into a dysbiotic one. The exact mechanisms by which these pathogens reorganize the healthy oral microbiome are still unknown. In the present manuscript, we present results demonstrating that P. gingivalis induces S. mitis death and DNA fragmentation in an in vitro biofilm system. Moreover, we report here the induction of expression of multiple transposases in a Streptococcus mitis biofilm when the periodontopathogen P. gingivalis is present. Based on these results, we hypothesize that P. gingivalis induces S. mitis cell death by an unknown mechanism, shaping the oral microbiome to its advantage. PMID:24866802

Duran-Pinedo, Ana E; Baker, Vinesha D; Frias-Lopez, Jorge

2014-08-01

97

A member of the peptidase M48 superfamily of Porphyromonas gingivalis is associated with virulence in vitro and in vivo  

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Full Text Available Background: In vivo-induced antigen technology was previously used to identify 115 genes induced in Porphyromonas gingivalis W83 during human infection. One of these, PG2197, a conserved hypothetical protein which has homology to a Zn-dependent protease, was examined with respect to a role in disease. Design: The expression of PG2197 in human periodontitis patients was investigated, but as there is increasing evidence of a direct relationship between P. gingivalis and cardiovascular disease, a mutation was constructed in this gene to also determine its role in adherence, invasion, and persistence within human coronary artery endothelial cells (HCAEC and neutrophil killing susceptibility. Results: Plaque samples from 20 periodontitis patients were analyzed by real-time PCR, revealing that PG2197 was expressed in 60.0% of diseased sites compared to 15.8% of healthy sites, even though P. gingivalis was detected in equal numbers from both sites. The expression of this gene was also found to be up-regulated in microarrays at 5 and 30 min of invasion of HCAEC. Interestingly, a PG2197 mutant displayed increased adherence, invasion, and persistence within HCAEC when compared to the wild-type strain. Conclusion: This gene appears to be important for the virulence of P. gingivalis, both in vivo and in vitro.

Sheila Walters

2009-11-01

98

Biochemical characterization of the ?-carbonic anhydrase from the oral pathogen Porphyromonas gingivalis, PgiCA.  

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Carbonic anhydrases (CAs, EC 4.2.1.1) catalyze a simple but physiologically relevant reaction in all life kingdoms, carbon dioxide hydration to bicarbonate and protons. CAs are present in many pathogenic species and are involved in the bicarbonate metabolism/biosynthetic reactions involving this ion. Ubiquity of these enzymes suggests a pivotal role in microbial virulence and pathogenicity. Porphyromonas gingivalis is an anaerobic bacterium, which colonizes the oral cavity, being involved in the pathogenesis of periodontitis, an inflammatory disease leading to tooth loss. Recently, we reported an anion inhibitory study on the ?-CA (denominated PgiCA) identified in the genome of this Gram-negative bacterium. In this paper we continue our research on PgiCA, and describe the biochemical characterization of the recombinant protein, its thermal stability, the oligomeric state and the enzyme kinetics. PgiCA is a polypeptide chain formed of 192 amino acids and displays an identity of 30-33% when compared with the prototypical ?-CAs, CAM or CAMH (from Methanosarcina thermophila) or CcmM (from Thermosynechococcus elongatus). A subunit molecular mass of 21 kDa was estimated by SDS-PAGE, while HPLC size exclusion chromatography under native conditions gave an estimated molecular mass of 65 kDa suggesting that the recombinant enzyme self-associate in a homotrimer, as all other ?-CAs studied so far. Enzyme kinetic analysis showed that PgiCA is 62 times more effective as a catalyst compared to CAM, the only other ?-CA characterized in detail kinetically. All these features represent an interesting attractive for the drug design of inhibitors/activators of this new enzyme. PMID:23914926

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

2014-08-01

99

Histidine kinase-mediated production and autoassembly of Porphyromonas gingivalis fimbriae.  

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Porphyromonas gingivalis, a Gram-negative oral anaerobe, is strongly associated with chronic adult periodontitis, and it utilizes FimA fimbriae to persistently colonize and evade host defenses in the periodontal crevice. The FimA-related gene cluster (the fim gene cluster) is positively regulated by the FimS-FimR two-component system. In this study, comparative analyses between fimbriate type strain ATCC 33277 and fimbria-deficient strain W83 revealed differences in their fimS loci, which encode FimS histidine kinase. Using a reciprocal gene exchange system, we established that FimS from W83 is malfunctional. Complementation analysis with chimeric fimS constructs revealed that W83 FimS has a defective kinase domain due to a truncated conserved G3 box motif that provides an ATP-binding pocket. The introduction of the functional fimS from 33277 restored the production, but not polymerization, of endogenous FimA subunits in W83. Further analyses with a fimA-exchanged W83 isogenic strain showed that even the fimbria-deficient W83 retains the ability to polymerize FimA from 33277, indicating the assembly of mature FimA by a primary structure-dependent mechanism. It also was shown that the substantial expression of 33277-type FimA fimbriae in the W83 derivative requires the introduction and expression of the functional 33277 fimS. These findings indicate that FimSR is the unique and universal regulatory system that activates the fim gene cluster in a fimA genotype-independent manner. PMID:20118268

Nishikawa, Kiyoshi; Duncan, Margaret J

2010-04-01

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

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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; LØbner, Morten

2009-01-01

 
 
 
 
101

Characterization of Wheat Germ Agglutinin Lectin-Reactive Glycosylated OmpA-Like Proteins Derived from Porphyromonas gingivalis.  

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Glycosylation is one of the common posttranslational modifications in eukaryotes. Recently, glycosylated proteins have also been identified in prokaryotes. A few glycosylated proteins, including gingipains, have been identified in Porphyromonas gingivalis, a major pathogen associated with chronic periodontitis. However, no other glycosylated proteins have been found. The present study identified glycoproteins in P. gingivalis cell lysates by lectin blotting. Whole-cell lysates reacted with concanavalin A (ConA), Lens culinaris agglutinin (LCA), Phaseolus vulgaris erythroagglutinin (PHA-E4), and wheat germ agglutinin (WGA), suggesting the presence of mannose-, N-acetylgalactosamine-, or N-acetylglucosamine (GlcNAc)-modified proteins. Next, glycoproteins were isolated by ConA-, LCA-, PHA-E4-, or WGA-conjugated lectin affinity chromatography although specific proteins were enriched only by the WGA column. Mass spectrometry analysis showed that an OmpA-like, heterotrimeric complex formed by Pgm6 and Pgm7 (Pgm6/7) was the major glycoprotein isolated from P. gingivalis. Deglycosylation experiments and Western blotting with a specific antibody indicated that Pgm6/7 was modified with O-GlcNAc. When whole-cell lysates from P. gingivalis mutant strains with deletions of Pgm6 and Pgm7 were applied to a WGA column, homotrimeric Pgm7, but not Pgm6, was isolated. Heterotrimeric Pgm6/7 had the strongest affinity for fibronectin of all the extracellular proteins tested, whereas homotrimeric Pgm7 showed reduced binding activity. These findings suggest that the heterotrimeric structure is important for the biological activity of glycosylated WGA-binding OmpA-like proteins in P. gingivalis. PMID:25135681

Murakami, Yukitaka; Hasegawa, Yoshiaki; Nagano, Keiji; Yoshimura, Fuminobu

2014-11-01

102

Involvement of PG2212 Zinc Finger Protein in the Regulation of Oxidative Stress Resistance in Porphyromonas gingivalis W83.  

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The adaptation of Porphyromonas gingivalis to H2O2-induced stress while inducible is modulated by an unknown OxyR-independent mechanism. Previously, we reported that the PG_2212 gene was highly upregulated in P. gingivalis under conditions of prolonged oxidative stress. Because this gene may have regulatory properties, its function in response to H2O2 was further characterized. PG2212, annotated as a hypothetical protein of unknown function, is a 10.3-kDa protein with a cysteine 2-histidine 2 (Cys2His2) zinc finger domain. The isogenic mutant P. gingivalis FLL366 (?PG_2212) showed increased sensitivity to H2O2 and decreased gingipain activity compared to the parent strain. Transcriptome analysis of P. gingivalis FLL366 revealed that approximately 11% of the genome displayed altered expression (130 downregulated genes and 120 upregulated genes) in response to prolonged H2O2-induced stress. The majority of the modulated genes were hypothetical or of unknown function, although some are known to participate in oxidative stress resistance. The promoter region of several of the most highly modulated genes contained conserved motifs. In electrophoretic mobility shift assays, the purified rPG2212 protein did not bind its own promoter region but bound a similar region in several of the genes modulated in the PG_2212-deficient mutant. A metabolome analysis revealed that PG2212 can regulate a number of genes coding for proteins involved in metabolic pathways critical for its survival under the conditions of oxidative stress. Collectively, our data suggest that PG2212 is a transcriptional regulator that plays an important role in oxidative stress resistance and virulence regulation in P. gingivalis. PMID:25225267

Dou, Yuetan; Aruni, Wilson; Luo, Tianlong; Roy, Francis; Wang, Charles; Fletcher, Hansel M

2014-12-01

103

Inhibitory Effect of Dodonaea viscosa var. angustifolia on the Virulence Properties of the Oral Pathogens Streptococcus mutans and Porphyromonas gingivalis.  

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Aim. This study investigated the effect of Dodonaea viscosa var. angustifolia (DVA) on the virulence properties of cariogenic Streptococcus mutans and Porphyromonas gingivalis implicated in periodontal diseases. Methods. S. mutans was cultured in tryptone broth containing a crude leaf extract of DVA for 16 hours, and the pH was measured after 10, 12, 14, and 16?h. Biofilms of S. mutans were grown on glass slides for 48 hours and exposed to plant extract for 30 minutes; the adherent cells were reincubated and the pH was measured at various time intervals. Minimum bactericidal concentration of the extracts against the four periodontal pathogens was determined. The effect of the subinhibitory concentration of plant extract on the production of proteinases by P. gingivalis was also evaluated. Results. DVA had no effect on acid production by S. mutans biofilms; however, it significantly inhibited acid production in planktonic cells. Periodontal pathogens were completely eliminated at low concentrations ranging from 0.09 to 0.02?mg/mL of crude plant extracts. At subinhibitory concentrations, DVA significantly reduced Arg-gingipain (24%) and Lys-gingipain (53%) production by P. gingivalis (P ? 0.01). Conclusions. These results suggest that DVA has the potential to be used to control oral infections including dental caries and periodontal diseases. PMID:24223061

Patel, Mrudula; Naidoo, Roxanne; Owotade, Foluso John

2013-01-01

104

Increased levels of Porphyromonas gingivalis are associated with ischemic and hemorrhagic cerebrovascular disease in humans: an in vivo study  

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Full Text Available OBJECTIVE: This study investigated the role of periodontal disease in the development of stroke or cerebral infarction in patients by evaluating the clinical periodontal conditions and the subgingival levels of periodontopathogens. MATERIAL AND METHODS: Twenty patients with ischemic (I-CVA or hemorrhagic (H-CVA cerebrovascular episodes (test group and 60 systemically healthy patients (control group were evaluated for: probing depth, clinical attachment level, bleeding on probing and plaque index. Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans were both identified and quantified in subgingival plaque samples by conventional and real-time PCR, respectively. RESULTS: The test group showed a significant increase in each of the following parameters: pocket depth, clinical attachment loss, bleeding on probing, plaque index and number of missing teeth when compared to control values (p<0.05, unpaired t-test. Likewise, the test group had increased numbers of sites that were contaminated with P. gingivalis (60%x10%; p<0.001; chi-squared test and displayed greater prevalence of periodontal disease, with an odds ratio of 48.06 (95% CI: 5.96-387.72; p<0.001. Notably, a positive correlation between probing depth and the levels of P. gingivalis in ischemic stroke was found (r=0.60; p=0.03; Spearman's rank correlation coefficient test. A. actinomycetemcomitans DNA was not detected in any of the groups by conventional or real-time PCR. CONCLUSIONS: Stroke patients had deeper pockets, more severe attachment loss, increased bleeding on probing, increased plaque indexes, and in their pockets harbored increased levels of P. gingivalis. These findings suggest that periodontal disease is a risk factor for the development of cerebral hemorrhage or infarction. Early treatment of periodontitis may counteract the development of cerebrovascular episodes.

Janaina Salomon Ghizoni

2012-02-01

105

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

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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

106

Increased levels of Porphyromonas gingivalis are associated with ischemic and hemorrhagic cerebrovascular disease in humans: an in vivo study  

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Full Text Available SciELO Brazil | Language: English Abstract in english OBJECTIVE: This study investigated the role of periodontal disease in the development of stroke or cerebral infarction in patients by evaluating the clinical periodontal conditions and the subgingival levels of periodontopathogens. MATERIAL AND METHODS: Twenty patients with ischemic (I-CVA) or hemor [...] rhagic (H-CVA) cerebrovascular episodes (test group) and 60 systemically healthy patients (control group) were evaluated for: probing depth, clinical attachment level, bleeding on probing and plaque index. Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans were both identified and quantified in subgingival plaque samples by conventional and real-time PCR, respectively. RESULTS: The test group showed a significant increase in each of the following parameters: pocket depth, clinical attachment loss, bleeding on probing, plaque index and number of missing teeth when compared to control values (p

Janaina Salomon, Ghizoni; Luís Antônio de Assis, Taveira; Gustavo Pompermaier, Garlet; Marcos Flávio, Ghizoni; Jefferson Ricardo, Pereira; Thiago José, Dionísio; Daniel Thomas, Brozoski; Carlos Ferreira, Santos; Adriana Campos Passanezi, Sant' Ana.

107

Integrin ?5?1-fimbriae binding and actin rearrangement are essential for Porphyromonas gingivalis invasion of osteoblasts and subsequent activation of the JNK pathway  

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Full Text Available Abstract Background Chronic periodontitis is an infectious disease of the periodontium, which includes the gingival epithelium, periodontal ligament and alveolar bone. The signature clinical feature of periodontitis is resorption of alveolar bone and subsequent tooth loss. The Gram-negative oral anaerobe, Porphyromonas gingivalis, is strongly associated with periodontitis, and it has been shown previously that P. gingivalis is capable of invading osteoblasts in a dose- and time-dependent manner resulting in inhibition of osteoblast differentiation and mineralization in vitro. It is not yet clear which receptors and cytoskeletal components mediate the invasive process, nor how the signaling pathways and viability of osteoblasts are affected by bacterial internalization. This study aimed to investigate these issues using an in vitro model system involving the inoculation of P. gingivalis ATCC 33277 into primary osteoblast cultures. Results It was found that binding between P. gingivalis fimbriae and integrin ?5?1 on osteoblasts, and subsequent peripheral condensation of actin, are essential for entry of P. gingivalis into osteoblasts. The JNK pathway was activated in invaded osteoblasts, and apoptosis was induced by repeated infections. Conclusions These observations indicate that P. gingivalis manipulates osteoblast function to promote its initial intracellular persistence by prolonging the host cell life span prior to its intercellular dissemination via host cell lysis. The identification of molecules critical to the interaction between P. gingivalis and osteoblasts will facilitate the development of new therapeutic strategies for the prevention of periodontal bone loss.

Zhang Wenjian

2013-01-01

108

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  

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Full Text Available SciELO Brazil | Language: English 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 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. 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 i [...] mplant 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.

Eduardo Augusto, Pfau; Mario Julio, Avila-Campos.

2005-09-01

109

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  

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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

2005-09-01

110

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 SciELO Brazil | Language: English 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 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. 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 i [...] mplant 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.

Eduardo Augusto, Pfau; Mario Julio, Avila-Campos.

111

Anion inhibition study of the ?-class carbonic anhydrase (PgiCAb) from the oral pathogen Porphyromonas gingivalis.  

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The oral pathogenic bacterium Porphyromonas gingivalis, encodes for two carbonic anhydrase (CA, EC 4.2.1.1) one belonging to the ?-class (PgiCAb) and another one to the ?-class (PgiCA). This last enzyme has been characterized earlier for its inhibition profile with various classes of CA inhibitors, such as the sulfonamides and anions, whereas for PgiCAb such data were not yet reported. Here we show that PgiCAb has a good catalytic activity for the CO2 hydration reaction, with kcat 2.8×10(5)s(-1) and kcat/Km of 1.5×10(7)M(-1)×s(-1), being inhibited by cyanate and diethyldithiocarbamate in the submillimolar range (KIs of 0.23-0.76mM) and more efficiently by sulfamide, sulfamate, phenylboronic acid and phenylarsonic acid (KIs of 60-78?M). The anion inhibition profile of the two P. gingivalis enzymes is very different. Identification of selective inhibitors of PgiCAb/PgiCA may lead to pharmacological tools useful for understanding the physiological role(s) of these enzymes, since this bacterium is the main causative agent of periodontitis and few treatment options are presently available. PMID:25168748

Vullo, Daniela; Del Prete, Sonia; Osman, Sameh M; Scozzafava, Andrea; Alothman, Zeid; Supuran, Claudiu T; Capasso, Clemente

2014-09-15

112

Improved Multiplex PCR Using Conserved and Species-Specific 16S rRNA Gene Primers for Simultaneous Detection of Actinobacillus actinomycetemcomitans, Bacteroides forsythus, and Porphyromonas gingivalis  

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Among putative periodontal pathogens, Actinobacillus actinomycetemcomitans, Bacteroides forsythus, and Porphyromonas gingivalis are most convincingly implicated as etiological agents in periodontitis. Therefore, techniques for detection of those three species would be of value. We previously published a description of a multiplex PCR that detects A. actinomycetemcomitans and P. gingivalis. The present paper presents an improvement on that technique, which now allows more sensitive detection of all three periodontal pathogens. Sensitivity was determined by testing serial dilutions of A. actinomycetemcomitans, B. forsythus, and P. gingivalis cells. Primer specificity was tested against (i) all gene sequences from the GenBank-EMBL database, (ii) six A. actinomycetemcomitans, one B. forsythus, and four P. gingivalis strains, (iii) eight different species of oral bacteria, and (iv) supra- and subgingival plaque samples from 20 healthy subjects and subgingival plaque samples from 10 patients with periodontitis. The multiplex PCR had a detection limit of 10 A. actinomycetemcomitans, 10 P. gingivalis, and 100 B. forsythus cells. Specificity was confirmed by the fact that (i) none of our forward primers were homologous to the 16S rRNA genes of other oral species, (ii) amplicons of predicted size were detected for all A. actinomycetemcomitans, B. forsythus, and P. gingivalis strains tested, and (iii) no amplicons were detected for the eight other bacterial species. A. actinomycetemcomitans, B. forsythus, and P. gingivalis were detected in 6 of 20, 1 of 20, and 11 of 20 of supragingival plaque samples, respectively, and 4 of 20, 7 of 20, and 13 of 20 of subgingival plaque samples, respectively, from periodontally healthy subjects. Among patients with periodontitis, the organisms were detected in 7 of 10, 10 of 10, and 7 of 10 samples, respectively. The simultaneous detection of three periodontal pathogens is an advantage of this technique over conventional PCR assays. PMID:10523542

Tran, Simon Dangtuan; Rudney, Joel D.

1999-01-01

113

In-Vivo Effect of Andrographolide on Alveolar Bone Resorption Induced by Porphyromonas gingivalis and Its Relation with Antioxidant Enzymes  

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Alveolar bone resorption is one of the most important facts in denture construction. Porphyromonas gingivalis (Pg) causes alveolar bone resorption, and morphologic measurements are the most frequent methods to identify bone resorption in periodontal studies. This study has aimed at evaluating the effect of Andrographolide (AND) on alveolar bone resorption in rats induced by Pg. 24 healthy male Sprague Dawley rats were divided into four groups as follows: normal control group and three experimental groups challenged orally with Pg ATCC 33277 five times a week supplemented with 20?mg/kg and 10?mg/kg of AND for twelve weeks. Alveolar bones of the left and right sides of the mandible were assessed by a morphometric method. The bone level, that is, the distance from the alveolar bone crest to cementumenamel junction (CEJ), was measured using 6.1?:?1 zoom stereomicroscope and software. AND reduced the effect of Pg on alveolar bone resorption and decreased the serum levels of Hexanoyl-Lysine (HEL); furthermore the reduced glutathione/oxidised glutathione (GSH/GSSG) ratio in AND treated groups (10 and 20?mg/kg) significantly increased when compared with the Pg group (P < 0.05). We can conclude that AND suppresses alveolar bone resorption caused by Pg in rats. PMID:24151590

Al Batran, Rami; Al-Bayaty, Fouad H.; Al-Obaidi, Mazen M. Jamil

2013-01-01

114

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

2010-09-01

115

Histatin 5 binds to Porphyromonas gingivalis hemagglutinin B (HagB) and alters HagB-induced chemokine responses  

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Histatins are human salivary gland peptides with anti-microbial and anti-inflammatory activities. In this study, we hypothesized that histatin 5 binds to Porphyromonas gingivalis hemagglutinin B (HagB) and attenuates HagB-induced chemokine responses in human myeloid dendritic cells. Histatin 5 bound to immobilized HagB in a surface plasmon resonance (SPR) spectroscopy-based biosensor system. SPR spectroscopy kinetic and equilibrium analyses, protein microarray studies, and I-TASSER structural modeling studies all demonstrated two histatin 5 binding sites on HagB. One site had a stronger affinity with a KD1 of 1.9 ?M and one site had a weaker affinity with a KD2 of 60.0 ?M. Binding has biological implications and predictive modeling studies and exposure of dendritic cells both demonstrated that 20.0 ?M histatin 5 attenuated (p < 0.05) 0.02 ?M HagB-induced CCL3/MIP-1?, CCL4/MIP-1?, and TNF? responses. Thus histatin 5 is capable of attenuating chemokine responses, which may help control oral inflammation.

Borgwardt, Derek S.; Martin, Aaron D.; van Hemert, Jonathan R.; Yang, Jianyi; Fischer, Carol L.; Recker, Erica N.; Nair, Prashant R.; Vidva, Robinson; Chandrashekaraiah, Shwetha; Progulske-Fox, Ann; Drake, David; Cavanaugh, Joseph E.; Vali, Shireen; Zhang, Yang; Brogden, Kim A.

2014-01-01

116

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

2010-01-01

117

The vimE Gene Downstream of vimA Is Independently Expressed and Is Involved in Modulating Proteolytic Activity in Porphyromonas gingivalis W83  

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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 ...

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

2004-01-01

118

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

2012-01-01

119

Myxomavirus Anti-Inflammatory Chemokine Binding Protein Reduces the Increased Plaque Growth Induced by Chronic Porphyromonas gingivalis Oral Infection after Balloon Angioplasty Aortic Injury in Mice  

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Thrombotic occlusion of inflammatory plaque in coronary arteries causes myocardial infarction. Treatment with emergent balloon angioplasty (BA) and stent implant improves survival, but restenosis (regrowth) can occur. Periodontal bacteremia is closely associated with inflammation and native arterial atherosclerosis, with potential to increase restenosis. Two virus-derived anti-inflammatory proteins, M-T7 and Serp-1, reduce inflammation and plaque growth after BA and transplant in animal models through separate pathways. M-T7 is a broad spectrum C, CC and CXC chemokine-binding protein. Serp-1 is a serine protease inhibitor (serpin) inhibiting thrombotic and thrombolytic pathways. Serp-1 also reduces arterial inflammation and improves survival in a mouse herpes virus (MHV68) model of lethal vasculitis. In addition, Serp-1 demonstrated safety and efficacy in patients with unstable coronary disease and stent implant, reducing markers of myocardial damage. We investigate here the effects of Porphyromonas gingivalis, a periodontal pathogen, on restenosis after BA and the effects of blocking chemokine and protease pathways with M-T7 and Serp-1. ApoE?/? mice had aortic BA and oral P. gingivalis infection. Arterial plaque growth was examined at 24 weeks with and without anti-inflammatory protein treatment. Dental plaques from mice infected with P. gingivalis tested positive for infection. Neither Serp-1 nor M-T7 treatment reduced infection, but IgG antibody levels in mice treated with Serp-1 and M-T7 were reduced. P. gingivalis significantly increased monocyte invasion and arterial plaque growth after BA (Pinfection. Blockade of chemokines, but not serine proteases significantly reduced arterial plaque growth, suggesting a central role for chemokine-mediated inflammation after BA in P. gingivalis infected mice. PMID:25354050

Lucas, Alexandra R.; Verma, Raj K.; Dai, Erbin; Liu, Liying; Chen, Hao; Kesavalu, Sheela; Rivera, Mercedes; Velsko, Irina; Ambadapadi, Sriram; Chukkapalli, Sasanka; Kesavalu, Lakshmyya

2014-01-01

120

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 SciELO Chile | Language: Spanish 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 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 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.

 
 
 
 
121

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 SciELO Chile | Language: Spanish 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 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 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.

2011-08-01

122

Prevalence of Enterococcus faecalis and Porphyromonas gingivalis in infected root canals and their susceptibility to endodontic treatment procedures: A molecular study  

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Full Text Available Introduction. Because apical periodontitis is recognizably an infectious disease, elimination or reduction of intracanal bacteria is of utmost importance for optimum treatment outcome. Objective. The prevalence of Enterococcus faecalis and Porphyromonas gingivalis in infected root canals was studied Also, the effect of endodontic therapy by using intracanal medicaments, calcium hydroxide paste (CH or gutta-percha points containing calcium hydroxide (CH-GP or chlorhexidine (CHX-GP on these microorganisms was assessed by polymerase chain reaction (PCR assay. Methods. Fifty-one patients with chronic apical periodontitis were randomly allocated in one of the following groups according to the intracanal medicament used: CH, CH-GP and CHX-GP group. Bacterial samples were taken upon access (S1, after chemomechanical instrumentation (S2 and after 15-day medication (S3. PCR assay was used to detect the presence of selected bacteria. Results. E. faecalis was detected in 49% (25/51 and P. gingivalis in 17.6% (9/51 of the samples. Samples which showed no bacterial presence at S1 were excluded from further analysis. Overall analysis of all 29 samples revealed significant differences between S1 and S2 (p<0.001, S2 and S3 (p<0.05, and S1 and S3 (p<0.001. When distinction was made between the intracanal medications, there was a significant difference in the number of PCR positive samples between S1 and S2, S1 and S3, but not between S2 and S3 samples. Conclusion. E. faecalis is more prevalent than P. gingivalis in primary endodontic infection. Intracanal medication in conduction with instrumentation and irrigation efficiently eliminates E. faecalis and P. gingivalis from infected root canals.

Stojanovi? Nikola

2014-01-01

123

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

2011-08-01

124

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

2013-07-15

125

Aerosolized clindamycin is superior to aerosolized dexamethasone or clindamycin-dexamethasone combination in the treatment of severe Porphyromonas gingivalis aspiration pneumonia in an experimental murine model.  

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Adjunctive corticosteroid treatment to reduce excessive local inflammatory response in pneumonia is controversial. To study the effects of an early local adjunct dexamethasone treatment on the course of pneumonia and inflammatory/cytokine response, mice were intratracheally inoculated with live Porphyromonas gingivalis and treated with either clindamycin (C), dexamethasone (D), C+D combination, or were not treated (Pg). Six mice from each group were euthanized at 6, 24, 72, and 168 hours after inoculation. Levels of tumor necrosis factor (TNF)-?, soluble TNF-? receptors (sTNFR1 and sTNFR2), interleukin (IL)-1?, and IL-6 in the serum and lung-homogenate supernatant were determined. Lung samples were histopathologically assessed and all findings compared to those found in 24 sham-inoculated mice (phosphate-buffered saline [PBS]). Severe P. gingivalis-induced bronchopneumonia progressed from 24 hours, peaked at 72 hours, and resolved after 168 hours with changes in local and systemic cytokine levels. Clindamycin-treated mice developed only mild bronchopneumonia that resolved fast (72 hours) with an early (6-24 hours) normalization of local and systemic cytokine levels. Similar course of pneumonia and cytokine level changes were observed in mice treated with C+D, but later. Early (6-24 hours) local elevation of sTNFRs was observed in C and C+D groups of mice, whereas nontreated (Pg) mice had increased systemic sTNFRs. Severe bronchopneumonia with delayed resolution was observed in D-group mice, with an early local and systemic decrease in sTNFR1 and persistent elevation of local TNF-?. Clindamycin or a clindamycin-dexamethasone combination treatment significantly improves the course of P. gingivalis-aspiration pneumonia, but more so if clindamycin alone is used. A favorable course of pneumonia seems to be associated with an early elevation of sTNFRs and normalization of TNF-?. PMID:22149928

Nemec, Ana; Pavlica, Zlatko; Nemec-Svete, Alenka; Eržen, Damijan; Milutinovi?, Aleksandra; Petelin, Milan

2012-02-01

126

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 SciELO Chile | Language: Spanish 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 Porphyromo [...] nas 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. 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 P [...] orphyromonas 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.

2010-04-01

127

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 SciELO Chile | Language: Spanish 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 Porphyromo [...] nas 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. 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 P [...] orphyromonas 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.

128

Porphyromonas gingivalis-derived lysine gingipain enhances osteoclast differentiation induced by tumor necrosis factor-? and interleukin-1? but suppresses that by interleukin-17A: importance of proteolytic degradation of osteoprotegerin by lysine gingipain.  

Science.gov (United States)

Periodontitis is a chronic inflammatory disease accompanied by alveolar bone resorption by osteoclasts. Porphyromonas gingivalis, an etiological agent for periodontitis, produces cysteine proteases called gingipains, which are classified based on their cleavage site specificity (i.e. arginine (Rgps) and lysine (Kgps) gingipains). We previously reported that Kgp degraded osteoprotegerin (OPG), an osteoclastogenesis inhibitory factor secreted by osteoblasts, and enhanced osteoclastogenesis induced by various Toll-like receptor (TLR) ligands (Yasuhara, R., Miyamoto, Y., Takami, M., Imamura, T., Potempa, J., Yoshimura, K., and Kamijo, R. (2009) Lysine-specific gingipain promotes lipopolysaccharide- and active-vitamin D3-induced osteoclast differentiation by degrading osteoprotegerin. Biochem. J. 419, 159-166). Osteoclastogenesis is induced not only by TLR ligands but also by proinflammatory cytokines, including tumor necrosis factor-? (TNF-?), interleukin (IL)-1?, and IL-17A, in inflammatory conditions, such as periodontitis. Although Kgp augmented osteoclastogenesis induced by TNF-? and IL-1? in co-cultures of mouse osteoblasts and bone marrow cells, it suppressed that induced by IL-17A. In a comparison of proteolytic degradation of these cytokines by Kgp in a cell-free system with that of OPG, TNF-? and IL-1? were less susceptible, whereas IL-17A and OPG were equally susceptible to degradation by Kgp. These results indicate that the enhancing effect of Kgp on cytokine-induced osteoclastogenesis is dependent on the difference in degradation efficiency between each cytokine and OPG. In addition, elucidation of the N-terminal amino acid sequences of OPG fragments revealed that Kgp primarily cleaved OPG in its death domain homologous region, which might prevent dimer formation of OPG required for inhibition of receptor activator of nuclear factor ?B ligand. Collectively, our results suggest that degradation of OPG by Kgp is a crucial event in the development of osteoclastogenesis and bone loss in periodontitis. PMID:24755218

Akiyama, Tomohito; Miyamoto, Yoichi; Yoshimura, Kentaro; Yamada, Atsushi; Takami, Masamichi; Suzawa, Tetsuo; Hoshino, Marie; Imamura, Takahisa; Akiyama, Chie; Yasuhara, Rika; Mishima, Kenji; Maruyama, Toshifumi; Kohda, Chikara; Tanaka, Kazuo; Potempa, Jan; Yasuda, Hisataka; Baba, Kazuyoshi; Kamijo, Ryutaro

2014-05-30

129

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 SciELO Chile | Language: Spanish 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 afinidades 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, modulatio [...] n 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-gingipains (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.

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

2012-12-01

130

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 SciELO Chile | Language: Spanish 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 afinidades 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, modulatio [...] n 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-gingipains (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.

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

131

Binding of Porphyromonas gingivalis Fimbriae to Proline-Rich Glycoproteins in Parotid Saliva via a Domain Shared by Major Salivary Components  

Science.gov (United States)

Porphyromonas gingivalis, a putative periodontopathogen, can bind to human saliva through its fimbriae. We previously found that salivary components from the submandibular and sublingual glands bind to P. gingivalis fimbriae and that acidic proline-rich protein (PRP) and statherin function as receptor molecules for fimbriae. In this study, we investigated the fimbria-binding components in parotid saliva. Fractionated human parotid saliva by gel-filtration chromatography was immobilized onto nitrocellulose membranes for the overlay assay following sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The salivary components on the membrane were allowed to interact with fimbriae purified from P. gingivalis ATCC 33277, and the interacted fimbriae were probed with anti-fimbria antibodies. The fimbriae were shown to bind to two forms of proline-rich glycoproteins (PRGs) as well as to acidic PRPs and statherin. Moreover, fimbriae bound to several components of smaller molecular size which appeared to be acidic PRP variants and basic PRPs. Fimbriae bound strongly to the purified PRGs adsorbed onto hydroxyapatite (HAP) beads. In contrast, PRGs in solution failed to inhibit the fimbrial binding to the immobilized PRGs on the HAP beads. These findings suggest that the appearance of binding site(s) of PRGs can be ascribed to their conformational changes. We previously identified the distinct segments within PRP and statherin molecules that are involved in fimbrial binding. The peptides analogous to the binding regions of PRP and statherin (i.e., PRP-C and STN-C) markedly inhibit the binding of fimbriae to PRP and statherin immobilized on the HAP beads, respectively. The PRP-C significantly inhibited the binding of fimbriae to PRG-coated HAP beads as well as to PRP on HAP beads. The peptide did not affect the binding of fimbriae to statherin, whereas the STN-C showed no effect on the fimbrial binding to PRPs or PRGs. In the overlay assay, the PRP-C clearly diminished the interactions between the fimbriae and the various salivary components, including PRPs, the PRGs, and the components with smaller molecular sizes but not statherin. These results strongly suggest that fimbriae bind to salivary components (except statherin) via common peptide segments. It is also suggested that fimbriae bind to saliva through the two distinct binding domains of receptory salivary components: (i) PRGs and PRPs and (ii) statherin. PMID:9573091

Amano, Atsuo; Shizukuishi, Satoshi; Horie, Hiroshi; Kimura, Shigenobu; Morisaki, Ichijiro; Hamada, Shigeyuki

1998-01-01

132

Role of ghrelin in modulation of s-nitrosylation-Dependent akt inactivation induced in salivary gland acinar cells by porphyromonas gingivalis  

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Full Text Available Ghrelin, a peptide hormone, newly identified in oral mucosal tissue, has emerged re-cently as a principal modulator of the in-flammatory responses to bacterial infection through the regulation of nitric oxide syn-thase system. In this study, using rat sub-lingual salivary gland acinar cells, we report that lipopolysaccharide (LPS of periodon-topathic bacterium, P. gingivalis- induced enhancement in the activity of inducible ni-tric oxide synthase (iNOS was associated with the suppression in Akt kinase activity and the impairment in constitutive (c cNOS phosphorylation. Further, we show that the detrimental effect of the LPS on Akt activa-tion, manifested in the kinase protein S-nitrosylation and a decrease in its phos-phorylation at Ser473, was susceptible to suppression by iNOS inhibitor, 1400W. Moreover, we demonstrate that a peptide hormone, ghrelin, countered the LPS- induced changes in Akt activity and NOS system. This effect of ghrelin was reflected in the decreased in Akt S-nitrosylation and the increase in its phosphorylation at Ser473, as well as cNOS activation through phos-phorylation. Our findings suggest that P. gingivalis-induced up-regulation in iNOS leads to Akt kinase inactivation through S-nitrosylation that impacts cNOS activation through phosphorylation. We also show that the countering effect of ghrelin on P. gingivalis-induced disturbances in Akt ac-tivation are manifested in a decrease in the kinase S-nitrosylation and the increase in its phosphorylation.

Bronislaw L. Slomiany

2010-12-01

133

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

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Full Text Available SciELO Brazil | Language: English 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 fro [...] m permanent (n=3) and deciduous (n=2) teeth were established using an explant technique. After the fourth passage, fibroblasts were stimulated by increasing 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.

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

2013-04-01

134

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

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Full Text Available 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 increasing 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.

Carla Renata Sipert

2013-04-01

135

Sulfonamide inhibition study of the carbonic anhydrases from the bacterial pathogen Porphyromonas gingivalis: the ?-class (PgiCAb) versus the ?-class (PgiCA) enzymes.  

Science.gov (United States)

The oral pathogenic bacterium Porphyromonas gingivalis, encodes for two carbonic anhydrases (CAs, EC 4.2.1.1) one belonging to the ?-class (PgiCA) and another one to the ?-class (PgiCAb). This last enzyme has been cloned and characterized here for its inhibition profile with the main class of CA inhibitors, the sulfonamides. Many of the clinically used sulfonamides as well as simple aromatic/heterocyclic sulfonamides were ineffective as PgiCAb inhibitors whereas better inhibition was observed with simple derivatives such as sulfanilamide, metanilamide, 4-aminoalkylbenzenesulfonamides (KIs of 364-475nM). The halogenosulfanilamides incorporating heavy halogens, 4-hydroxy- and 4-hydroxyalkyl-benzenesulfonamides, were also micromolar, ineffective PgiCAb inhibitors. The best inhibitors of the ?-class enzyme were acetazolamide and ethoxzolamide, with KIs of 214-280nM. Interestingly, the ?-class enzyme was much more sensitive to sulfonamide inhibitors compared to the ?-class one, PgiCAb. Identification of potent and possibly selective inhibitors of PgiCAb/PgiCA may lead to pharmacological tools useful for understanding the physiological role(s) of these enzymes, since this bacterium is the main causative agent of periodontitis and few treatment options are presently available. PMID:25129169

Prete, Sonia Del; Vullo, Daniela; Osman, Sameh M; Scozzafava, Andrea; AlOthman, Zeid; Capasso, Clemente; Supuran, Claudiu T

2014-09-01

136

Site-specific O-Glycosylation on the MUC2 Mucin Protein Inhibits Cleavage by the Porphyromonas gingivalis Secreted Cysteine Protease (RgpB)  

DEFF Research Database (Denmark)

The colonic epithelial surface is protected by an inner mucus layer that the commensal microflora cannot penetrate. We previously demonstrated that Entamoeba histolytica secretes a protease capable of dissolving this layer that is required for parasite penetration. Here, we asked whether there are bacteria that can secrete similar proteases. We screened bacterial culture supernatants for such activity using recombinant fragments of the MUC2 mucin, the major structural component, and the only gel-forming mucin in the colonic mucus. MUC2 has two central heavily O-glycosylated mucin domains that are protease-resistant and has cysteine-rich N and C termini responsible for polymerization. Culture supernatants of Porphyromonas gingivalis, a bacterium that secretes proteases responsible for periodontitis, cleaved the MUC2 C-terminal region, whereas the N-terminal region was unaffected. The active enzyme was isolated and identified as Arg-gingipain B (RgpB). Two cleavage sites were localized to IR?TT and NR?QA. IR?TTcleavage will disrupt the MUC2 polymers. Because this site has two potential O-glycosylation sites, we tested whether recombinant GalNAc-transferases (GalNAc-Ts) could glycosylate a synthetic peptide covering the IRTT sequence. Only GalNAc-T3 was able to glycosylate the second Thr in IRTT, rendering the sequence resistant to cleavage by RgpB. Furthermore, when GalNAc-T3 was expressed in CHO cells expressing the MUC2 C terminus, the second threonine was glycosylated, and the protein became resistant to RgpB cleavage. These findings suggest that bacteria can produce proteases capable of dissolving the inner protective mucus layer by specific cleavages in the MUC2 mucin and that this cleavage can be modulated by site-specific O-glycosylation.

van der Post, Sjoerd; Subramani, Durai B

2013-01-01

137

The K1K2 region of Lys-gingipain of Porphyromonas gingivalis blocks induction of HLA expression by gamma interferon.  

Science.gov (United States)

In the context of periodontal disease, cysteine proteinases or gingipains from Porphyromonas gingivalis have been implicated in the hydrolysis of cytokines, including gamma interferon (IFN-?). This cytokine plays a crucial role in host defenses, in part, by regulating expression of major histocompatibility complex molecules. Our recent analysis has identified three structurally defined modules, K1, K2, and K3, of the hemagglutinin region of the lysine gingipain Kgp. These three structurally homologous domains have a common ?-sandwich topology that is similar to that found in a superfamily of adhesins and carbohydrate binding domains. The three Kgp hemagglutinin modules are distinguished by variation in some of the loops projecting from the ?-sandwich core. Recombinant products corresponding to both single and multidomain regions as well as native Kgp bound IFN-? with similar affinities. Among the adhesin domain constructs, only the K1K2 polypeptide inhibited the upregulation of HLA-1 expression in a human erythroleukemia (K562) line induced by both recombinant and native IFN-?. The K1K2 polypeptide also inhibited HLA-DR expression induced by IFN-? in human umbilical vein endothelial cells. These effects were competitively inhibited by coincubation with sodium or potassium chloride solution. The N-terminal residues of IFN-? were implicated in mediating the effect of K1K2, while antibody binding to loop 1 of K2 blocked the action of K1K2. The findings indicate the potential significance of structurally defined Kgp adhesin modules in the inactivation of IFN-? but also the potential of K1K2 in locating the target for the catalytic domain of Kgp. PMID:22802347

Yun, Peter L; Li, Nan; Collyer, Charles A; Hunter, Neil

2012-10-01

138

Expression levels of novel cytokine IL-32 in periodontitis and its role in the suppression of IL-8 production by human gingival fibroblasts stimulated with Porphyromonas gingivalis  

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Full Text Available Background:IL-32 was recently found to be elevated in the tissue of rheumatoid arthritis and inflammatory bowel disease. Periodontitis is a chronic inflammatory disease caused by polymicrobial infections that result in soft tissue destruction and alveolar bone loss. Although IL-32 is also thought to be associated with periodontal disease, its expression and possible role in periodontal tissue remain unclear. Therefore, this study investigated the expression patterns of IL-32 in healthy and periodontally diseased gingival tissue. The expression of IL-32 in cultured human gingival fibroblasts (HGF as well as effects of autocrine IL-32 on IL-8 production from HGF were also examined.Methods:Periodontal tissue was collected from both healthy volunteers and periodontitis patients, and immunofluorescent staining was performed in order to determine the production of IL-32. Using real-time PCR and ELISA, mRNA expression and protein production of IL-32 in HGF, stimulated by Porphyromonas gingivalis (Pg, were also investigated.Results:Contrary to our expectation, the production of IL-32 in the periodontitis patients was significantly lower than in the healthy volunteers. According to immunofluorescent microscopy, positive staining for IL-32 was detected in prickle and basal cell layers in the epithelium as well as fibroblastic cells in connective tissue. Addition of fixed Pg in vitro was found to suppress the otherwise constitutive expression of IL-32 mRNA and protein in HGF. However, recombinant IL-32 in vitro inhibited the expression of IL-8 mRNA by HGF stimulated with Pg. Interestingly, anti-IL-32 neutralizing antibody upregulated the IL-8 mRNA expression in non-stimulated HGF, indicating that constitutive expression of IL-32 in HGF suppressed IL-8 mRNA expression in the absence of bacterial stimulation.Conclusion:These results indicate that IL-32 is constitutively produced by HGF which can be suppressed by Pg and may play a role in the downregulation of inflammatory responses, such as IL-8 production, in periodontal tissue.

Kazuhisa Ouhara

2012-03-01

139

Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans IgG Subclass Antibody Levels as Immunological Risk Indicators of Chronic Periodontitis: A Multilevel Approach / Niveles de Anticuerpos Subclase IgG de Porphyromonas gingivalis y Aggregatibacter actinomycetemcomitans como Indicadores de Riesgo Inmunológico de Periodontitis Crónica: un Enfoque Multinivel  

Scientific Electronic Library Online (English)

Full Text Available SciELO Chile | Language: English Abstract in spanish Los niveles de anticuerpos en algunos patógenos periodontales están asociados con mayores niveles de marcadores inflamatorios. El propósito de este estudio fue examinar la contribución relativa de inmunoglobulina sérica G (IgG) factores de nivel de anticuerpos de subclase y factores locales en la pr [...] ofundidad del sondaje en periodontitis crónica. Se tomaron muestras de suero de 444 pacientes con diagnóstico de periodontitis moderada y grave y de 223 sujetos de control. Se determinaron los títulos de anticuerpos IgG subclase a Porphyromonas gingivalis (Pg), Aggregatibacter actinomycetemcomitans (Aa) y Tanerella forsythia (Tf) mediante inmunoensayo indirecto (ELISA). La contribución relativa de los pacientes, los dientes, y el sitio asociado a los parámetros en la profundidad de sondaje fueron evaluados con un modelo multinivel jerárquico. Los resultados indicaron que los pacientes con periodontitis tenían niveles detectables de IgG1 e IgG2. Altos niveles de anticuerpos IgG1 e IgG2 contra Aa fueron observados en 132 y 142 pacientes con periodontitis, respectivamente. Niveles altos de anticuerpos IgG1 e IgG2 contra Pg fueron detectados en 141 y 138 en pacientes con periodontitis respectivamente, y niveles altos de anticuerpos IgG1 e IgG2 contra Tf se produjeron en 121 y 136 pacientes con periodontitis, respectivamente. La mayor parte de la varianza se atribuyó a nivel de sitio (48%). El análisis multinivel asociados a profundidad de sondaje con factores relacionados a los sujetos, anticuerpos (suero IgG1 e IgG2 Aa y Pg), factores de los dientes (tipo) y los factores del sitio (localización mesial - distal y sangrado al sondaje). Anticuerpos elevados de suero IgG1 e IgG2 Aa y Pg (factores de los sujetos) reflejan el estado de la enfermedad periodontal destructiva. Abstract in english Antibody levels to some periodontal pathogens are associated with enhanced levels of inflammatory markers. The purpose of the current study was to examine the relative contribution of serum immunoglobulin G (IgG) subclass antibody level factors and local factors on the probing pocket depth in chroni [...] c periodontitis. Serum samples were taken from 444 patients diagnosed with moderate and severe periodontitis and 223 control subjects. The IgG subclass antibody titers to Porphyromonas gingivalis (Pg), Aggregatibacter actinomycetemcomitans (Aa) and Tanerella forsythia (Tf) using indirect immunoassay (ELISA) were determined. The relative contribution of patient, tooth and site-associated parameters on the probing pocket depth were evaluated with a hierarchical multilevel model. The results indicated that periodontitis patients had detectable levels of IgG1 and IgG2. High IgG1 and IgG2 antibody levels against Aa occurred in 132 and 142 periodontitis patients, respectively. High IgG1 and IgG2 antibody levels against Pg occurred in 141 and 138, periodontitis patients, respectively, and High IgG1 and IgG2 antibody levels against Tf occurred in 121 and 136 periodontitis patients, respectively. The majority of the variance was attributed to the site level (48%). The multilevel analysis associated deeper probing depth with subject factors (serum IgG1 and IgG2 antibody to Pg and Aa), tooth factors (tooth type), and site factors (mesial-distal location and bleeding on probing). Elevated serum IgG1 and IgG2 antibody to Pg and Aa (subject factors) reflects destructive periodontal disease status.

Carlos M, Ardila; Isabel C, Guzmán; Lyan, Bermudez; Sebastian, Bernau; Adolfo, Contreras; Andres, Duque; Sylvia, Duarte; Juliette, De Avila; Gloria Ines, Lafaurie.

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Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans IgG Subclass Antibody Levels as Immunological Risk Indicators of Chronic Periodontitis: A Multilevel Approach / Niveles de Anticuerpos Subclase IgG de Porphyromonas gingivalis y Aggregatibacter actinomycetemcomitans como Indicadores de Riesgo Inmunológico de Periodontitis Crónica: un Enfoque Multinivel  

Scientific Electronic Library Online (English)

Full Text Available SciELO Chile | Language: English Abstract in spanish Los niveles de anticuerpos en algunos patógenos periodontales están asociados con mayores niveles de marcadores inflamatorios. El propósito de este estudio fue examinar la contribución relativa de inmunoglobulina sérica G (IgG) factores de nivel de anticuerpos de subclase y factores locales en la pr [...] ofundidad del sondaje en periodontitis crónica. Se tomaron muestras de suero de 444 pacientes con diagnóstico de periodontitis moderada y grave y de 223 sujetos de control. Se determinaron los títulos de anticuerpos IgG subclase a Porphyromonas gingivalis (Pg), Aggregatibacter actinomycetemcomitans (Aa) y Tanerella forsythia (Tf) mediante inmunoensayo indirecto (ELISA). La contribución relativa de los pacientes, los dientes, y el sitio asociado a los parámetros en la profundidad de sondaje fueron evaluados con un modelo multinivel jerárquico. Los resultados indicaron que los pacientes con periodontitis tenían niveles detectables de IgG1 e IgG2. Altos niveles de anticuerpos IgG1 e IgG2 contra Aa fueron observados en 132 y 142 pacientes con periodontitis, respectivamente. Niveles altos de anticuerpos IgG1 e IgG2 contra Pg fueron detectados en 141 y 138 en pacientes con periodontitis respectivamente, y niveles altos de anticuerpos IgG1 e IgG2 contra Tf se produjeron en 121 y 136 pacientes con periodontitis, respectivamente. La mayor parte de la varianza se atribuyó a nivel de sitio (48%). El análisis multinivel asociados a profundidad de sondaje con factores relacionados a los sujetos, anticuerpos (suero IgG1 e IgG2 Aa y Pg), factores de los dientes (tipo) y los factores del sitio (localización mesial - distal y sangrado al sondaje). Anticuerpos elevados de suero IgG1 e IgG2 Aa y Pg (factores de los sujetos) reflejan el estado de la enfermedad periodontal destructiva. Abstract in english Antibody levels to some periodontal pathogens are associated with enhanced levels of inflammatory markers. The purpose of the current study was to examine the relative contribution of serum immunoglobulin G (IgG) subclass antibody level factors and local factors on the probing pocket depth in chroni [...] c periodontitis. Serum samples were taken from 444 patients diagnosed with moderate and severe periodontitis and 223 control subjects. The IgG subclass antibody titers to Porphyromonas gingivalis (Pg), Aggregatibacter actinomycetemcomitans (Aa) and Tanerella forsythia (Tf) using indirect immunoassay (ELISA) were determined. The relative contribution of patient, tooth and site-associated parameters on the probing pocket depth were evaluated with a hierarchical multilevel model. The results indicated that periodontitis patients had detectable levels of IgG1 and IgG2. High IgG1 and IgG2 antibody levels against Aa occurred in 132 and 142 periodontitis patients, respectively. High IgG1 and IgG2 antibody levels against Pg occurred in 141 and 138, periodontitis patients, respectively, and High IgG1 and IgG2 antibody levels against Tf occurred in 121 and 136 periodontitis patients, respectively. The majority of the variance was attributed to the site level (48%). The multilevel analysis associated deeper probing depth with subject factors (serum IgG1 and IgG2 antibody to Pg and Aa), tooth factors (tooth type), and site factors (mesial-distal location and bleeding on probing). Elevated serum IgG1 and IgG2 antibody to Pg and Aa (subject factors) reflects destructive periodontal disease status.

Carlos M, Ardila; Isabel C, Guzmán; Lyan, Bermudez; Sebastian, Bernau; Adolfo, Contreras; Andres, Duque; Sylvia, Duarte; Juliette, De Avila; Gloria Ines, Lafaurie.

2013-12-01

 
 
 
 
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Thiazolidinedione (pioglitazone) blocks P. gingivalis- and F. nucleatum, but not E. coli, lipopolysaccharide (LPS)-induced interleukin-6 (IL-6) production in adipocytes.  

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An elevated level of C-reactive protein (CRP) predicts the future development of coronary heart disease. Periodontitis appears to up-regulate CRP. CRP is produced by hepatocytes in response to interleukin-6 (IL-6). A major source of IL-6 in obese subjects is adipocytes. We hypothesized that lipopolysaccharide (LPS) from periodontal pathogens stimulated adipocytes to produce IL-6, and that the production was suppressed by the drugs targeted against insulin resistance, thiazolidinedione (pioglitazone), since this agent potentially showed an anti-inflammatory effect. Mouse 3T3-L1 adipocytes were stimulated with E. coli, P. gingivalis, and F. nucleatum LPS. The IL-6 concentration in culture supernatants was measured. All LPS stimulated adipocytes to produce IL-6. Although pioglitazone changed adipocyte appearance from large to small, and completely suppressed P. gingivalis and F. nucleatum LPS-induced IL-6 production, E. coli LPS-induced IL-6 production was not efficiently blocked. Thus, pioglitazone completely blocked periodontal-bacteria-derived LPS-induced IL-6 production in adipocytes, a major inducer of CRP. PMID:15723863

Yamaguchi, M; Nishimura, F; Naruishi, H; Soga, Y; Kokeguchi, S; Takashiba, S

2005-03-01

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A combination of both arginine- and lysine-specific gingipain activity of Porphyromonas gingivalis is necessary for the generation of the micro-oxo bishaem-containing pigment from haemoglobin.  

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The black pigment of Porphyromonas gingivalis is composed of the mu-oxo bishaem complex of Fe(III) protoporphyrin IX (mu-oxo oligomer, dimeric haem), namely [Fe(III)PPIX]2O. P. gingivalis W50 and Rgp (Arg-gingipain)- and Kgp (Lys-gingipain)-deficient mutants K1A, D7, E8 and W501 [Aduse-Opoku, Davies, Gallagher, Hashim, Evans, Rangarajan, Slaney and Curtis (2000) Microbiology 146, 1933-1940] were grown on horse blood/agar for 14 days and examined for the production of mu-oxo bishaem. Mu-oxo Bishaem was detected by UV-visible, Mössbauer and Raman spectroscopies in wild-type W50 and in the black-pigmented RgpA- and RgpB-deficient mutants (W501 and D7 respectively), whereas no haem species were detected in the straw-coloured colonies of Kgp-deficient strain K1A. The dark brown pigment of the double RgpA/RgpB knockout mutant (E8) was not composed of mu-oxo bishaem, but of a high-spin monomeric Fe(III) protoporphyrin IX species (possibly a haem-albumin complex). In vitro incubation of oxyhaemoglobin with cells of the W50 strain and the RgpA- and RgpB-deficient mutants (W501 and D7) resulted in the formation of mu-oxo bishaem via methaemoglobin as an intermediate. Although the Kgp-deficient strain K1A converted oxyhaemoglobin into methaemoglobin, this was not further degraded into mu-oxo bishaem. The double RgpA/RgpB knockout was also not capable of producing mu-oxo bishaem from oxyhaemoglobin, but instead generated a haemoglobin haemichrome. Inhibition of Arg-X protease activity of W50, W501, D7 and K1A with leupeptin, under conditions where Lys-X protease activity was unaffected, prevented the production of mu-oxo bishaem from oxyhaemoglobin, but resulted in the formation of a haemoglobin haemichrome. These results show that one or both of RgpA and RgpB gingipains, in addition to the lysine-specific gingipain, is necessary for the production of mu-oxo bishaem from haemoglobin by whole cells of P. gingivalis. PMID:14741050

Smalley, John W; Thomas, Michael F; Birss, Andrew J; Withnall, Robert; Silver, Jack

2004-05-01

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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

2011-12-01

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LPS from P. gingivalis and Hypoxia Increases Oxidative Stress in Periodontal Ligament Fibroblasts and Contributes to Periodontitis  

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Oxidative stress is characterized by an accumulation of reactive oxygen species (ROS) and plays a key role in the progression of inflammatory diseases. We hypothesize that hypoxic and inflammatory events induce oxidative stress in the periodontal ligament (PDL) by activating NOX4. Human primary PDL fibroblasts were stimulated with lipopolysaccharide from Porphyromonas gingivalis (LPS-PG), a periodontal pathogen bacterium under normoxic and hypoxic conditions. By quantitative PCR, immunoblot, immunostaining, and a specific ROS assay we determined the amount of NOX4, ROS, and several redox systems. Healthy and inflamed periodontal tissues were collected to evaluate NOX4 and redox systems by immunohistochemistry. We found significantly increased NOX4 levels after hypoxic or inflammatory stimulation in PDL cells (P NOX4 and redox systems is crucial for ROS formation which plays a pivotal role during oral diseases. PMID:25374447

Golz, L.; Memmert, S.; Rath-Deschner, B.; Jager, A.; Appel, T.; Baumgarten, G.; Gotz, W.; Frede, S.

2014-01-01

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Inhibition of gingival fibroblast growth by Bacteroides gingivalis.  

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Human gingival fibroblasts were exposed in culture to cell extracts of different black-pigmented Bacteroides species, and their growth was monitored by determining thymidine uptake and counting cells. Of the Bacteroides species tested (B. gingivalis, B. asaccharolyticus, and B. intermedius), B. gingivalis gave the extract with the strongest inhibitory effect on fibroblast thymidine uptake. Linear inhibition reaching 80% of the control level was obtained with a dose of 100 micrograms of B. gingivalis extract protein per ml. The effect of B. asaccharolyticus resembled that of B. gingivalis, but even at the highest dose tested B. intermedius had only a slight inhibitory effect. When fibroblasts were counted after 2- and 4-day exposures to B. gingivalis extracts, a clear depression in the number of fibroblasts was found. The effects of extracts obtained from early and late growth phases of B. gingivalis cultures were similar. A fraction of B. gingivalis consisting essentially of lipopolysaccharides (LPSs) was obtained by degrading the extract proteins with proteinase K. Silver staining of polyacrylamide gels revealed a LPS pattern with a molecular mass ranging from 37 to 60 kilodaltons. This LPS-rich fraction caused inhibition of thymidine uptake by gingival fibroblasts similar to that caused by the native extract alone. Thus, the inhibition of gingival fibroblast growth by B. gingivalis appeared to be LPS mediated. This inhibitory effect of B. gingivalis on oral fibroblast growth may be a virulence factor of this bacterium. Images PMID:3793230

Larjava, H; Uitto, V J; Eerola, E; Haapasalo, M

1987-01-01

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Lipopolysaccharides from atherosclerosis-associated bacteria antagonize TLR4, induce formation of TLR2/1/CD36 complexes in lipid rafts and trigger TLR2-induced inflammatory responses in human vascular endothelial cells.  

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Infection with bacteria such as Chlamydia pneumonia, Helicobacter pylori or Porphyromonas gingivalis may be triggering the secretion of inflammatory cytokines that leads to atherogenesis. The mechanisms by which the innate immune recognition of these pathogens could lead to atherosclerosis remain unclear. In this study, using human vascular endothelial cells or HEK-293 cells engineered to express pattern-recognition receptors (PRRs), we set out to determine Toll-like receptors (TLRs) and functionally associated PRRs involved in the innate recognition of and response to lipopolysaccharide (LPS) from H. pylori or P. gingivalis. Using siRNA interference or recombinant expression of cooperating PRRs, we show that H. pylori and P. gingivalis LPS-induced cell activation is mediated through TLR2. Human vascular endothelial cell activation was found to be lipid raft-dependent and to require the formation of heterotypic receptor complexes comprising of TLR2, TLR1, CD36 and CD11b/CD18. In addition, we report that LPS from these bacterial strains are able to antagonize TLR4. This antagonistic activity of H. pylori or P. gingivalis LPS, as well as their TLR2 activation capability may be associated with their ability to contribute to atherosclerosis. PMID:17419716

Triantafilou, Martha; Gamper, Frederick G J; Lepper, Philipp M; Mouratis, Marios Angelos; Schumann, Christian; Harokopakis, Evlambia; Schifferle, Robert E; Hajishengallis, George; Triantafilou, Kathy

2007-08-01

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Effect of eel galectin AJL-1 on periodontopathic bacterial biofilm formation and their lipopolysaccharide-mediated inflammatory cytokine induction.  

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Porphyromonas gingivalis, Prevotella intermedia and Aggregatibacter actinomycetemcomitans, infectious pathogenic bacteria found in oral biofilm, cause periodontal disease. The inhibitory effect of AJL-1, a galectin present in the skin mucus of the Japanese eel Anguilla japonica, on biofilm formation by each of these strains was investigated by staining adherent bacteria on culture plates with crystal violet. An ATP bioluminescence assay was used to determine whether inhibition of biofilm formation was due to the bactericidal activity of AJL-1. The effect of AJL-1 on cytokine induction in human umbilical vascular endothelial cells (HUVECs) by lipopolysaccharide (LPS) isolated from A. actinomycetemcomitans was also investigated by enzyme-linked immunosorbent assay (ELISA). AJL-1 significantly inhibited biofilm formation by A. actinomycetemcomitans strains Y4, ATCC 29523 and ATCC 29524 but not by any strain of P. gingivalis or P. intermedia, and showed no bactericidal activity against A. actinomycetemcomitans strains. AJL-1 markedly suppressed interleukin (IL)-6 and IL-8 induction in HUVECs by LPS from A. actinomycetemcomitans strains Y4 and ATCC 29523. These observations indicate that AJL-1 is an effective inhibitor of biofilm formation by A. actinomycetemcomitans as well as of inflammatory cytokine induction in HUVECs by LPS. These finding indicate that AJL-1 may be of therapeutic value in A. actinomycetemcomitans-associated periodontal diseases. PMID:19505801

Takayama, Saori; Saitoh, Eiichi; Kimizuka, Ryuta; Yamada, Satoru; Kato, Tetsuo

2009-10-01

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Lysine substitutions convert a bacterial-agglutinating peptide into a bactericidal peptide that retains anti-lipopolysaccharide activity and low hemolytic activity.  

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GL13NH2 is a bacteria-agglutinating peptide derived from the sequence of the salivary protein parotid secretory protein (PSP, BPIFA2, SPLUNC2, C20orf70). The peptide agglutinates both Gram negative and Gram positive bacteria, and shows anti-lipopolysaccharide activity in vitro and in vivo. However, GL13NH2 does not exhibit bactericidal activity. To generate a more cationic peptide with potential bactericidal activity, three amino acid residues were replaced with lysine residues to generate the peptide GL13K. In this report, the antibacterial and anti-inflammatory activities of GL13K were characterized. GL13K had lost the ability to agglutinate bacteria but gained bactericidal activity. Substitution of individual amino acids in GL13K with alanine did not restore bacterial agglutination. GL13K was bactericidal against Pseudomonas aeruginosa, Streptococcus gordonii and Escherichia coli but not Porphyromonas gingivalis. Unlike the agglutinating activity of GL13NH2, the bactericidal activity of GL13K against P. aeruginosa was retained in the presence of saliva. Both GL13NH2 and GL13K exhibited anti-lipopolysaccharide activity. In GL13K, this activity appeared to depend on a serine hydroxyl group. GL13K protected mice from lipopolysaccharide-induced sepsis and the peptide exhibited a low level of hemolysis, suggesting that it may be suitable for in vivo application. PMID:22484285

Abdolhosseini, Mahsa; Nandula, Seshagiri R; Song, Jonathan; Hirt, Helmut; Gorr, Sven-Ulrik

2012-06-01

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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

150

Anti-inflammatory activity of fisetin in human gingival fibroblasts treated with lipopolysaccharide.  

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Fisetin is an anti-inflammatory flavonoid; however, its anti-inflammatory mechanism is not yet understood. In this study, we evaluated the anti-inflammatory effect of fisetin and its association with mitogen-activated protein kinase (MAPK) and nuclear factor kappa-beta pathways in human gingival fibroblasts (HGFs) treated with lipopolysaccharide (LPS) obtained from Porphyromonas gingivalis. The cell signaling, cell viability, and cyclooxygenase-2 (COX-2) expression of HGFs treated with various concentrations (0, 1, 5, 10, and 15 ?M) of fisetin were measured by cell viability assay (MTT), Western blotting, and reverse transcriptase polymerase chain reaction analysis on COX-2. We found that fisetin significantly reduced the synthesis and expression of prostaglandin E2 in HGFs treated with LPS. Activation of extracellular signal-regulated kinase, c-Jun N-terminal kinase, and p38 MAPK was suppressed consistently by fisetin in HGFs treated with LPS. The data indicate that fisetin inhibits MAPK activation and COX-2 expression without affecting cell viability. These findings may be valuable for understanding the mechanism of the effect of fisetin on periodontal disease. PMID:25263652

Gutiérrez-Venegas, Gloria; Contreras-Sánchez, Anabel; Ventura-Arroyo, Jairo Agustín

2014-10-01

151

Lysine-specific gingipain promotes lipopolysaccharide- and active vitamin D3-induced osteoclast differentiation by degrading osteoprotegerin  

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SYNOPSIS Porphyromonas gingivalis is one of the major pathogens of periodontitis, a condition characterized by excessive alveolar bone resorption by osteoclasts. The bacterium produces cysteine proteases called gingipains, which are classified according to their cleavage-site specificity into lysine-specific (Kgp) and arginine-specific gingipains (Rgps). In this study, we examined the effects of gingipains on osteoclast differentiation. In co-cultures of mouse bone marrow cells and osteoblasts, formation of multinucleated osteoclasts induced by 1?,25-dihydroxyvitamin D3 [1?,25(OH)2D3] was augmented by Kgp but not by RgpB. A physiological concentration (0.1 nM) of 1?,25(OH)2D3 induced the osteoclast formation in the presence of 100 nM Kgp to the extent comparable to that induced by 10 nM 1?,25(OH)2D3. Kgp also enhanced osteoclastogenesis induced by various microbial components including lipopolysaccharide. Combined use of Kgp and 1?,25(OH)2D3 or lipopolysaccharide also increased the number of resorption pits developed on dentin slices, indicating the osteoclasts formed in the presence of Kgp possess bone-resorbing activity. The enhanced osteoclastogenesis by Kgp was correlated with a depletion of osteoprotegerin in co-culture media and proteolytic activity-dependent, since benzyloxycarbonyl-phenylalanyl-lysyl-acycloxyketone, an inhibitor of Kgp, completely abolished osteoclastogenesis induced by Kgp. Kgp digested osteoprotegerin, since its recombinant protein was susceptible to degradation by Kgp in the presence of serum. As a result, Kgp did not augment osteoclastogenesis in co-cultures of osteoprotegerin-deficient osteoblasts and bone marrow cells. In addition, enhanced osteoclastogenesis by Kgp was abolished by excess amount of recombinant osteoprotegerin. These findings suggest that degradation of osteoprotegerin is one of the mechanisms underlying promotion of osteoclastogenesis by Kgp. PMID:19102726

Yasuhara, Rika; Miyamoto, Yoichi; Takami, Masamichi; Imamura, Takahisa; Potempa, Jan; Yoshimura, Kentaro; Kamijo, Ryutaro

2014-01-01

152

Oral P. gingivalis infection alters the vascular reactivity in healthy and spontaneously atherosclerotic mice  

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Full Text Available Abstract Background Considering that recent studies have demonstrated endothelial dysfunction in subjects with periodontitis and that there is no information about vascular function in coexistence of periodontitis and atherosclerosis, we assessed the impact of oral inoculation with the periodontal pathogen Porphyromonas gingivalis on vascular reactivity in healthy and hypercholesterolemic apolipoprotein E-deficient (ApoE mice. In vitro preparations of mesenteric arteriolar bed were used to determine the vascular responses to acetylcholine, sodium nitroprusside and phenylephrine (PE. Results Alveolar bone resorption, an evidence of periodontitis, was assessed and confirmed in all infected mice. Acetylcholine- and sodium nitroprusside-induced vasorelaxations were similar among all groups. Non-infected ApoE mice were hyperreactive to PE when compared to non-infected healthy mice. P gingivalis infection significantly enhanced the vasoconstriction to PE in both healthy and spontaneous atherosclerotic mice, when compared to their respective controls. Conclusions This study demonstrates that oral P gingivalis affects the alpha-adrenoceptor-mediated vascular responsiveness in both healthy and spontaneous atherosclerotic mice, reinforcing the association between periodontitis and cardiovascular diseases.

Stefanon Ivanita

2011-05-01

153

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

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Full Text Available SciELO Brazil | Language: English 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), 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

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

154

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

2009-07-01

155

Structural Modifications of Bacterial Lipopolysaccharide that Facilitate Gram-Negative Bacteria Evasion of Host Innate Immunity.  

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Bacterial lipopolysaccharide (LPS), a cell wall component characteristic of Gram-negative bacteria, is a representative pathogen-associated molecular pattern that allows mammalian cells to recognize bacterial invasion and trigger innate immune responses. The polysaccharide moiety of LPS primary plays protective roles for bacteria such as prevention from complement attacks or camouflage with common host carbohydrate residues. The lipid moiety, termed lipid A, is recognized by the Toll-like receptor 4 (TLR4)/MD-2 complex, which transduces signals for activation of host innate immunity. The basic structure of lipid A is a glucosamine disaccharide substituted by phosphate groups and acyl groups. Lipid A with six acyl groups (hexa-acylated form) has been indicated to be a strong stimulator of the TLR4/MD-2 complex. This type of lipid A is conserved among a wide variety of Gram-negative bacteria, and those bacteria are easily recognized by host cells for activation of defensive innate immune responses. Modifications of the lipid A structure to less-acylated forms have been observed in some bacterial species, and those forms are poor stimulators of the TLR4/MD-2 complex. Such modifications are thought to facilitate bacterial evasion of host innate immunity, thereby enhancing pathogenicity. This hypothesis is supported by studies of Yersinia pestis LPS, which contains hexa-acylated lipid A when the bacterium grows at 27°C (the temperature of the vector flea), and shifts to contain less-acylated forms when grown at the human body temperature of 37°C. This alteration of lipid A forms following transmission of Y. pestis from fleas to humans contributes predominantly to the virulence of this bacterium over other virulence factors. A similar role for less-acylated lipid A forms has been indicated in some other bacterial species, such as Francisella tularensis, Helicobacter pylori, and Porphyromonas gingivalis, and further studies to explore this concept are expected. PMID:23745121

Matsuura, Motohiro

2013-01-01

156

Induction of heme oxygenase-1 attenuates lipopolysaccharide-induced inflammasome activation in human gingival epithelial cells.  

Science.gov (United States)

Interleukin-1? (IL-1?) is a pathogenic factor for the destruction of periodontal tissue in periodontitis. The processing of IL-1? is regulated by cytosolic machinery termed as the inflammasome, which recruits and activates caspase-1 and then cleaves pro-IL-1? to produce mature IL-1?. Porphyromonas gingivalis (P. gingivalis) infection and lipopolysaccharide (LPS) have been shown to activate the NLRP3 inflammasome and stimulate IL-1? production in human oral cells. Heme oxygenase-1 (HO-1) is an ubiquitous cytoprotective enzyme. The products of HO-1 exhibit protective biological activities, including antioxidant and anti-inflammatory effects. In the present study, we investigated the hypothesis that the induction of HO-1 inhibits the activation of the inflammasome and protects against LPS-induced inflammatory damage in cultured human gingival epithelial cells (GECs). Our results revealed that LPS induced the overexpression of the inflammasome components, NLRP3, apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) and caspase-1, by western blot analysis. Co-immunoprecipitation analysis indicated that LPS increased the binding of NLRP3 and ASC, and confocal imaging revealed that LPS increased the immunostaining and co-localization of ASC and caspase-1, indicating that LPS enhanced the assembly/formation of the inflammasome components. Hemin, a potent HO-1 inducer, blocked the LPS-induced overexpression and the formation of the NLRP3 inflammasome. Furthermore, hemin also inhibited the LPS-induced increase in the production of IL-1?, as shown by enzyme-linked immunosorbent assay and blocked the nuclear translocation of pro-inflammatory nuclear factor-?B (NF-?B), as shown by confocal assays. As a result, hemin protected the cells from LPS-induced damage, which was demonstrated by the immunostaining pattern of the cell junction protein, E-cadherin. LPS produced a disturbed staining pattern of E-cadherin, suggesting the disruption of epithelial integrity, which was abolished in the hemin-treated cells. In conclusion, our data demonstrate that the induction of HO-1 by hemin attenuates LPS-induced inflammatory damage in human GECs through the inhibition of inflammasome activation. PMID:25069505

Li, Hong; Zhou, Xiaoli; Zhang, Jianli

2014-10-01

157

Rapid Myeloid Cell Transcriptional and Proteomic Responses to Periodontopathogenic Porphyromonas gingivalis  

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Long-lived monocytes, macrophages, and dendritic cells (DCs) are Toll-like receptor-expressing, antigen-presenting cells derived from a common myeloid lineage that play key roles in innate and adaptive immune responses. Based on immunohistochemical and molecular analyses of inflamed tissues from patients with chronic destructive periodontal disease, these cells, found in the inflammatory infiltrate, may drive the progressive periodontal pathogenesis. To investigate early transcriptional signa...

Nares, Salvador; Moutsopoulos, Niki M.; Angelov, Nikola; Rangel, Zoila G.; Munson, Peter J.; Sinha, Neha; Wahl, Sharon M.

2009-01-01

158

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

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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

2010-06-01

159

Genetic transformation of an obligate anaerobe, P. gingivalis for FMN-green fluorescent protein expression in studying host-microbe interaction.  

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The recent introduction of "oxygen-independent" flavin mononucleotide (FMN)-based fluorescent proteins (FbFPs) is of major interest to both eukaryotic and prokaryotic microbial biologists. Accordingly, we demonstrate for the first time that an obligate anaerobe, the successful opportunistic pathogen of the oral cavity, Porphyromonas gingivalis, can be genetically engineered for expression of the non-toxic green FbFP. The resulting transformants are functional for studying dynamic bacterial processes in living host cells. The visualization of the transformed P. gingivalis (PgFbFP) revealed strong fluorescence that reached a maximum emission at 495 nm as determined by fluorescence microscopy and spectrofluorometry. Human primary gingival epithelial cells (GECs) were infected with PgFbFP and the bacterial invasion of host cells was analyzed by a quantitative fluorescence microscopy and antibiotic protection assays. The results showed similar levels of intracellular bacteria for both wild type and PgFbFP strains. In conjunction with organelle specific fluorescent dyes, utilization of the transformed strain provided direct and accurate determination of the live/metabolically active P. gingivalis' trafficking in the GECs over time. Furthermore, the GECs were co-infected with PgFbFP and the ATP-dependent Clp serine protease-deficient mutant (ClpP-) to study the differential fates of the two strains within the same host cells. Quantitative co-localization analyses displayed the intracellular PgFbFP significantly associated with the endoplasmic reticulum network, whereas the majority of ClpP- organisms trafficked into the lysosomes. Hence, we have developed a novel and reliable method to characterize live host cell-microbe interactions and demonstrated the adaptability of FMN-green fluorescent protein for studying persistent host infections induced by obligate anaerobic organisms. PMID:21525983

Choi, Chul Hee; DeGuzman, Jefferson V; Lamont, Richard J; Yilmaz, Özlem

2011-01-01

160

Inhibition of gingival fibroblast growth by Bacteroides gingivalis.  

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Human gingival fibroblasts were exposed in culture to cell extracts of different black-pigmented Bacteroides species, and their growth was monitored by determining thymidine uptake and counting cells. Of the Bacteroides species tested (B. gingivalis, B. asaccharolyticus, and B. intermedius), B. gingivalis gave the extract with the strongest inhibitory effect on fibroblast thymidine uptake. Linear inhibition reaching 80% of the control level was obtained with a dose of 100 micrograms of B. gin...

Larjava, H.; Uitto, V. J.; Eerola, E.; Haapasalo, M.

1987-01-01

 
 
 
 
161

Prevalence of actinobacillus actinomycetemcomitans and prophyromonas gingivalis in subgingival microflora of patients with aggressive periodontitis  

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Full Text Available Statement of Problem: One of the best ways for treatment of Aggressive Periodontitis (AP is identification and elimination of etiologic factors specially two microorganisms Actinobacillus actinomycetemcomitans (Aa and Porphyromonas gingivalis (Pg in patients harboring them. Purpose: This study determines the prevalence of Aa and Pg and its correlation with age, sex and the number of family members as well as probing pocket depth (PPD in active sites of AP patients, referred to department of periodontics, Faculty of Dentistry, Tehran University of Medical Sciences. Materials and Methods: In this cross sectional, descriptive study, 54 sites (PPD> 5mm in 15 patients were considered for culture. Marginal gingiva was dried and sampling performed by paperpoint (#30. The selective medium for Aa, was Trypticase Soy Agar-Bacitracin- Vancomycin (TSBV and for Pg was Brucella agar. Results were analyzed using Fisher and Chi-Square statistical tests. Results: Thirteen patients or 38 sites (70.4% were identified as Aa positive and 3 patients or 10 sites (18.4% were Pg positive. There was no significant relation between the presence of Aa and sex or age (P=0.086. Pg was more prevalent in men compared with women (P<0.0001 but with regard to age there was no statistical difference between men and women. Aa had a significant positive correlation with PPD (P=0.002, which was not true for Pg. In addition, the number of positive sites showed a significant negative correlation with the number of family members. Conclusion: Based on the present study, the prevalence of Aa in deep pockets in patients with AP is higher than Pg.

Paknejad M.

2005-05-01

162

Effect of Allium cepa L. on Lipopolysaccharide-Stimulated Osteoclast Precursor Cell Viability, Count, and Morphology Using 4?,6-Diamidino-2-phenylindole-Staining  

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Allium cepa L. is known to possess numerous pharmacological properties. Our aim was to examine the in vitro effects of Allium cepa L. extract (AcE) on Porphyromonas gingivalis LPS and Escherichia coli LPS-stimulated osteoclast precursor cells to determine cell viability to other future cell-based assays. Osteoclast precursor cells (RAW 264.7) were stimulated by Pg LPS (1??g/mL) and E. coli LPS (1??g/mL) in the presence or absence of different concentrations of AcE (10–1000??g/mL) for 5 days at 37°C/5% CO2. Resazurin reduction and total protein content assays were used to detect cell viability. AcE did not affect cell viability. Resazurin reduction assay showed that AcE, at up to 1000??g/mL, did not significantly affect cell viability and cellular protein levels. Additionally a caspase 3/7 luminescence assay was used to disclose apoptosis and there was no difference in apoptotic activity between tested groups and control group. Fluorescence images stained by DAPI showed no alteration on the morphology and cell counts of LPS-stimulated osteoclast precursor cells with the use of AcE in all tested concentrations when compared to control. These findings suggest that Allium cepa L. extract could be used for in vitro studies on Porphyromonas gingivalis LPS and Escherichia coli LPS-stimulated osteoclast precursor cells. PMID:25221602

Oliveira, Tatiane; Figueiredo, Camila A.; Stavroullakis, Alexander; Da Silva Velozo, Eudes; Nogueira-Filho, Getulio

2014-01-01

163

Fluorescence immunoassay for detecting periodontal bacterial pathogens in plaque.  

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A particle concentration fluorescence immunoassay has been modified into a bacterial concentration fluorescence immunoassay (BCFIA) to rapidly detect periodontopathic bacteria in human plaque samples. The BCFIA utilizes fluorescently tagged monoclonal antibodies (MAbs) directed against the lipopolysaccharide of selected gram-negative plaque bacteria. Microorganisms closely associated with periodontal disease that can be identified in plaque with the BCFIA include Porphyromonas gingivalis, Bac...

Wolff, L. F.; Anderson, L.; Sandberg, G. P.; Aeppli, D. M.; Shelburne, C. E.

1991-01-01

164

Pathogenicity of Porphyromonas asaccharolytica in Adult Periodontitis of Malaysian Samples  

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Full Text Available Bacterial toxin of Porphyromonas asaccharolytica strains obtained from adult periodontitis were analysed using mouse model. Results from subcutaneous injection with whole bacterial cells of 108 and 1010 cells mL-1 failed to induce any infection. However when bacterial concentration of 1012 cells mL-1 was used, all P. asaccharolytica strains were able to induced the development of localized lesion on balb/c mice. In addition, infected mice appeared cachectic with ruffled hair. Thus, infection with the latter concentration was pathogenic and was dose dependent. The toxin was also found to be heat labile for all strains. Therefore, we conclude that the extracellularly toxin produced by P. asaccharolytica of our local isolates which were obtained from adult periodontitis, is virulent and heat labile.

W.H. Himratul-Aznita

2008-01-01

165

El Lipopolisacárido / Lipopolysaccharide  

Scientific Electronic Library Online (English)

Full Text Available SciELO Colombia | Language: Spanish Abstract in spanish El lipopolisacárido (LPS) o endotoxina es el mayor componente de la membrana externa de las bacterias Gram negativas, desempeñan una importante función en la activación del sistema inmune al constituir el antígeno superficial más importante de este tipo de bacterias. El LPS está compuesto por una re [...] gión lípidica y una glicosídica con funciones separadas y/o sinérgicas lo que hace de esta molécula uno de los factores de virulencia más complejos de comprender, esta revisión pretende hacer un acercamiento para dimensionar la universalidad y diversidad de efectos del principal responsable del shock endotóxico inducido por bacterias Gram negativas Abstract in english The lipopolysaccharide (LPS) or endotoxin is the major component of the outer membrane in Gram negative bacterial pathogens; it plays an important role in the activation of the immune system to be the most important surface antigen of Gram negative bacteria. The LPS consist of a lipophilic component [...] and a polysaccharide portion with different function that makes this molecule one of the virulence factors more complex to understand, this review make an approach to measure the diversity effects and universality of LPS

Stefany, Romero Hurtado; Carlos Arturo, Iregui.

166

First Evidence of Genetic Intraspecific Variability and Occurrence of Entamoeba gingivalis in HIV(+)/AIDS  

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Entamoeba gingivalis is considered an oral commensal but demonstrates a pathogenic potential associated with periodontal disease in immunocompromised individuals. Therefore, this study evaluated the occurrence, opportunistic conditions, and intraspecific genetic variability of E. gingivalis in HIV(+)/AIDS patients. Entamoeba gingivalis was studied using fresh examination (FE), culture, and PCR from bacterial plaque samples collected from 82 HIV(+)/AIDS patients. Genetic characterization of the lower ribosomal subunit of region 18S (18S-SSU rRNA) was conducted in 9 positive samples using low-stringency single specific primer PCR (LSSP-PCR) and sequencing analysis. Entamoeba gingivalis was detected in 63.4% (52/82) of the samples. No association was detected between the presence of E. gingivalis and the CD4+ lymphocyte count (?200 cells/mm3 (p?=?0.912) or viral load (p?=?0.429). The LSSP-PCR results helped group E. gingivalis populations into 2 polymorphic groups (68.3% similarity): group I, associated with 63.6% (7/11) of the samples, and group II, associated with 36.4% (4/11) of the samples, which shared 74% and 83.7% similarity and association with C and E isolates from HIV(?) individuals, respectively. Sequencing of 4 samples demonstrated 99% identity with the reference strain ATCC 30927 and also showed 2 divergent clusters, similar to those detected by LSSP-PCR. Opportunistic behavior of E. gingivalis was not detected, which may be related to the use of highly active antiretroviral therapy by all HIV(+)/AIDS patients. The high occurrence of E. gingivalis in these patients can be influenced by multifactorial components not directly related to the CD4+ lymphocyte counts, such as cholesterol and the oral microbiota host, which could mask the potential opportunistic ability of E. gingivalis. The identification of the 18S SSU-rRNA polymorphism by LSSP-PCR and sequencing analysis provides the first evidence of genetic variability in E. gingivalis isolated from HIV patients. PMID:24376598

Cembranelli, Sibeli B. S.; Souto, Fernanda O.; Ferreira-Paim, Kennio; Richinho, Tulio T.; Nunes, Poliana L.; Nascentes, Gabriel A. N.; Ferreira, Thatiana B.; Correia, Dalmo; Lages-Silva, Eliane

2013-01-01

167

First evidence of genetic intraspecific variability and occurrence of Entamoeba gingivalis in HIV(+)/AIDS.  

Science.gov (United States)

Entamoeba gingivalis is considered an oral commensal but demonstrates a pathogenic potential associated with periodontal disease in immunocompromised individuals. Therefore, this study evaluated the occurrence, opportunistic conditions, and intraspecific genetic variability of E. gingivalis in HIV(+)/AIDS patients. Entamoeba gingivalis was studied using fresh examination (FE), culture, and PCR from bacterial plaque samples collected from 82 HIV(+)/AIDS patients. Genetic characterization of the lower ribosomal subunit of region 18S (18S-SSU rRNA) was conducted in 9 positive samples using low-stringency single specific primer PCR (LSSP-PCR) and sequencing analysis. Entamoeba gingivalis was detected in 63.4% (52/82) of the samples. No association was detected between the presence of E. gingivalis and the CD4(+) lymphocyte count (?200 cells/mm(3) (p?=?0.912) or viral load (p?=?0.429). The LSSP-PCR results helped group E. gingivalis populations into 2 polymorphic groups (68.3% similarity): group I, associated with 63.6% (7/11) of the samples, and group II, associated with 36.4% (4/11) of the samples, which shared 74% and 83.7% similarity and association with C and E isolates from HIV(-) individuals, respectively. Sequencing of 4 samples demonstrated 99% identity with the reference strain ATCC 30927 and also showed 2 divergent clusters, similar to those detected by LSSP-PCR. Opportunistic behavior of E. gingivalis was not detected, which may be related to the use of highly active antiretroviral therapy by all HIV(+)/AIDS patients. The high occurrence of E. gingivalis in these patients can be influenced by multifactorial components not directly related to the CD4(+) lymphocyte counts, such as cholesterol and the oral microbiota host, which could mask the potential opportunistic ability of E. gingivalis. The identification of the 18S SSU-rRNA polymorphism by LSSP-PCR and sequencing analysis provides the first evidence of genetic variability in E. gingivalis isolated from HIV patients. PMID:24376598

Cembranelli, Sibeli B S; Souto, Fernanda O; Ferreira-Paim, Kennio; Richinho, Túlio T; Nunes, Poliana L; Nascentes, Gabriel A N; Ferreira, Thatiana B; Correia, Dalmo; Lages-Silva, Eliane

2013-01-01

168

Bacteroides gingivalis antigens and bone resorbing activity in root surface fractions of periodontally involved teeth  

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Bone resorbing activity and the presence of antigens of Bacteroides gingivalis were assessed in plaque, calculus, cementum, and dentin obtained from roots of teeth previously exposed to periodontitis. Each fraction was obtained by scaling the root surface. The fraction were extracted by stirring and sonication, and the soluble centrifuged, sterilized, dialyzed, and adjusted to equivalent protein concentrations. Cementum and dentin extracts from impacted teeth were prepared similarly and served as controls. Stimulation of bone resorption by each extract was assessed in organ cultures of fetal rat bones by measurement of release of previously-incorporated /sup 45/Ca from the bone into the medium. In some groups of teeth, calculus and cementum were treated with acid prior to scaling. Citric acid washes were recovered and dialyzed. An enzyme-linked immunosorbent assay (ELISA) was used to assess the extracts for the presence of antigens reactive with an antiserum to B. gingivalis. Significant stimulation of bone resorption was found in all calculus and periodontally-involved cementum preparations. ELISA showed significant levels of B.gingivalis antigens in plaque, calculus, and cementum of periodontally-involved teeth, but not in involved dentin nor in cementum or dentin of impact teeth. Treatment with citric acid removed essentially all B.gingivalis antigens from cementum but not calculus. The results suggest that substances which stimulate bone resorption and substances which react with B. gingivalis antiserum are present in surface plaque, calculus, and cementum or periodontally-involved teeth. These substances are not present in cementum and dentin of impacted teeth nor in dentin of periodontally-involved teeth. Treatment by both scaling and citric demineralization will remove most of these substances from cementum of teeth previously exposed to periodontitis.

Patters, M.R.; Landsberg, R.L.; Johansson, L.A.; Trummel, C.L.; Robertson, P.R. (Department of Periodontology, University of Connecticut, School of Dental Medicine, Farmington, Connecticut, U.S.A.)

1982-01-01

169

Bacteroides gingivalis antigens and bone resorbing activity in root surface fractions of periodontally involved teeth  

International Nuclear Information System (INIS)

Bone resorbing activity and the presence of antigens of Bacteroides gingivalis were assessed in plaque, calculus, cementum, and dentin obtained from roots of teeth previously exposed to periodontitis. Each fraction was obtained by scaling the root surface. The fraction were extracted by stirring and sonication, and the soluble centrifuged, sterilized, dialyzed, and adjusted to equivalent protein concentrations. Cementum and dentin extracts from impacted teeth were prepared similarly and served as controls. Stimulation of bone resorption by each extract was assessed in organ cultures of fetal rat bones by measurement of release of previously-incorporated 45Ca from the bone into the medium. In some groups of teeth, calculus and cementum were treated with acid prior to scaling. Citric acid washes were recovered and dialyzed. An enzyme-linked immunosorbent assay (ELISA) was used to assess the extracts for the presence of antigens reactive with an antiserum to B. gingivalis. Significant stimulation of bone resorption was found in all calculus and periodontally-involved cementum preparations. ELISA showed significant levels of B.gingivalis antigens in plaque, calculus, and cementum of periodontally-involved teeth, but not in involved dentin nor in cementum or dentin of impact teeth. Treatment with citric acid removed essentially all B.gingivalis antigens from cementum but not calculus. The results suggest that substances which stimulate bone resorption and substawhich stimulate bone resorption and substances which react with B. gingivalis antiserum are present in surface plaque, calculus, and cementum or periodontally-involved teeth. These substances are not present in cementum and dentin of impacted teeth nor in dentin of periodontally-involved teeth. Treatment by both scaling and citric demineralization will remove most of these substances from cementum of teeth previously exposed to periodontitis. (author)

170

The distribution pattern of Halicephalobus gingivalis in a horse is suggestive of a haematogenous spread of the nematode.  

Science.gov (United States)

The majority of Halicephalobus gingivalis-infections in horses have been fatal and are usually not diagnosed before necropsy. Therefore, knowledge about the nematode and the pathogenesis of infection in horses is limited. This has resulted in an on-going discussion about the port of entry and subsequent dissemination of H. gingivalis within the host. The present case of H. gingivalis-infection in a horse was diagnosed ante mortem. Post mortem findings, the distribution pattern of H. gingivalis nematodes in the brain, a high prevalence of inflammation in close relation to blood vessels, and the presence of the nematode in multiple organs with a disseminated pattern of distribution strongly suggested a haematogenous spread of the nematode in the horse. PMID:25233889

Henneke, Christina; Jespersen, Anna; Jacobsen, Stine; Nielsen, Martin K; McEvoy, Fintan; Jensen, Henrik E

2014-01-01

171

Multivariate analyses of cellular fatty acids in Bacteroides, Prevotella, Porphyromonas, Wolinella, and Campylobacter spp.  

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The genera Bacteroides, Wolinella, and Campylobacter contain several similar species that require taxonomic revision. Fatty acid profiles of whole bacterial cells have proven useful for taxonomy. In this study, cellular fatty acids from Bacteroides, Prevotella, Porphyromonas, Wolinella, and Campylobacter spp. were identified and quantitated by gas chromatography and gas chromatography-mass spectrometry, and the data were subjected to principal component analyses. Bacteroides fragilis, the typ...

Brondz, I.; Olsen, I.

1991-01-01

172

Biosynthesis of mycobacterial methylglucose lipopolysaccharides.  

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Mycobacterial pathogenesis is closely associated with a unique cell envelope rich in complex carbohydrates and unique lipids, among which are the mycolic acids. Mycobacteria also synthesize unique intracellular polymethylated polysaccharides (PMPSs), namely methylglucose lipopolysaccharides (MGLPs), which are acylated with short-chain fatty acids, and methylmannose polysaccharides (MMPs). Since PMPSs modulate the synthesis of long-chain fatty acids in vitro, the possibility of a similar role in vivo and the regulation of mycolic acids assembly have been anticipated. Unlike MGLPs, MMPs have been identified in M. smegmatis and other fast-growing mycobacteria but not in M. tuberculosis, implying an essential role for MGLPs in this pathogen and turning the biosynthetic enzymes into attractive drug targets. The genome of M. tuberculosis was decoded 14 years ago but only recently has the identity of the genes involved in MGLPs biosynthesis been investigated. Two gene clusters (Rv1208-Rv1213 and Rv3030-Rv3037c) containing a few genes considered to be essential for M. tuberculosis growth, have initially been proposed to coordinate MGLPs biosynthesis. Among these genes, only the product of Rv1208 for the first step in the MGLPs pathway has, so far, been crystallized and its three-dimensional structure been determined. However, recent results indicate that at least three additional clusters may be involved in this pathway. The functional assignment of authentic roles to some of these M. tuberculosis H37Rv genes sheds new light on the intricacy of MGLPs biogenesis and renewed interest on their biological role. PMID:22678749

Mendes, Vitor; Maranha, Ana; Alarico, Susana; Empadinhas, Nuno

2012-08-01

173

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

2013-08-01

174

Multivariate analyses of cellular fatty acids in Bacteroides, Prevotella, Porphyromonas, Wolinella, and Campylobacter spp.  

Science.gov (United States)

The genera Bacteroides, Wolinella, and Campylobacter contain several similar species that require taxonomic revision. Fatty acid profiles of whole bacterial cells have proven useful for taxonomy. In this study, cellular fatty acids from Bacteroides, Prevotella, Porphyromonas, Wolinella, and Campylobacter spp. were identified and quantitated by gas chromatography and gas chromatography-mass spectrometry, and the data were subjected to principal component analyses. Bacteroides fragilis, the type species of the genus Bacteroides, was distinct from the other organisms. While Bacteroides gracilis, Wolinella succinogenes, Wolinella curva, Wolinella recta, and Campylobacter fetus subsp. venerealis were close to each other, Prevotella (Bacteroides) buccae, Prevotella oralis, Prevotella oris, Prevotella disiens, Prevotella veroralis, Prevotella heparinolyticus, Porphyromonas (Bacteroides) endodontalis, and Bacteroides ureolyticus could be distinguished. B. fragilis was characterized by the presence of C3OH-i-1-, Ca-15, and Ci-15 and the absence of C12:0 and unsaturated fatty acids. For comparison, B. gracilis, B. ureolyticus, W. succinogenes, W. curva, W. recta, and Campylobacter fetus subsp. venerealis contained C12:0, C16:1, C18:1, and C3-OH-14 acids but lacked branched hydroxy and branched nonhydroxy acids. B. gracilis and B. ureolyticus are not "true" bacteroides. PMID:1993755

Brondz, I; Olsen, I

1991-01-01

175

The plant coumarins auraptene and lacinartin as potential multifunctional therapeutic agents for treating periodontal disease  

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Full Text Available Abstract Background Periodontal diseases are bacterial infections leading to chronic inflammation disorders that are frequently observed in adults. In the present study, we evaluated the effect of auraptene and lacinartin, two natural oxyprenylated coumarins, on the growth, adherence properties, and collagenase activity of Porphyromonas gingivalis. We also investigated the capacity of these compounds to reduce cytokine and matrix metalloproteinase (MMP secretion by lipopolysaccharide (LPS-stimulated macrophages and to inhibit MMP-9 activity. Methods Microplate dilution assays were performed to determine the effect of auraptene and lacinartin on P. gingivalis growth as well as biofilm formation stained with crystal violet. Adhesion of FITC-labeled P. gingivalis to oral epithelial cells was monitored by fluorometry. The effects of auraptene and lacinartin on LPS-induced cytokine and MMP secretion by macrophages were determined by immunological assays. Fluorogenic assays were used to evaluate the capacity of the two coumarins to inhibit the activity of P. gingivalis collagenase and MMP-9. Results Only lacinartin completely inhibited P. gingivalis growth in a complex culture medium. However, under iron-limiting conditions, auraptene and lacinartin both inhibited the growth of P. gingivalis. Lacinartin also inhibited biofilm formation by P. gingivalis and promoted biofilm desorption. Both compounds prevented the adherence of P. gingivalis to oral epithelial cells, dose-dependently reduced the secretion of cytokines (IL-8 and TNF-? and MMP-8 and MMP-9 by LPS-stimulated macrophages, and inhibited MMP-9 activity. Lacinartin also inhibited P. gingivalis collagenase activity. Conclusions By acting on multiple targets, including pathogenic bacteria, tissue-destructive enzymes, and the host inflammatory response, auraptene and lacinartin may be promising natural compounds for preventing and treating periodontal diseases.

Marquis Annie

2012-06-01

176

Nontoxic lipopolysaccharide from Rhodopseudomonas sphaeroides ATCC 17023.  

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Chemical analysis of the lipopolysaccharide from Rhodopseudomonas sphaeroides ATCC 17023, isolated by the phenol-chloroform-petroleum ether method, revealed the presence of glucuronic acid, 2-keto-3-deoxyoctonate, threonine, and phosphorus in the polysaccharide moiety. The lipid A component contained glucosamine, glucosamine phosphate, amide-bound 3-oxotetradecanoic acid and 3-hydroxytetradecanoic acid, and ester-bound 3-hydroxydecanoic acid and 7-tetradecenoic acid. Structural similarity of ...

Strittmatter, W.; Weckesser, J.; Salimath, P. V.; Galanos, C.

1983-01-01

177

Estudos de freqüência, morfologia e diagnóstico de Entamoeba gingivalis, Gros, 1849  

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Full Text Available Realizamos estudos de freqüência de Entamoeba gingivalis entre 100 pacientes atendidos nos ambulatórios odontológicos da Ufiiversidade Federal de Uberlândia (UFU, utilizando-se esfregaços corados pela técnica de Papanicolaou modificado, revelando um expressivo índice de 62% de positividade. A afinidade do corante pelo conteúdo vacuolarfagocítico impede uma nítida visualização das cromatinas central e periférica do núcleo do parasita. Lavados bucais de outros 10 pacientes foram utilizados para avaliar em qual método parasitológico de diagnóstico (a fresco e em coloração por hematoxilina férrica, Giemsa e Papanicolaou ocorre melhor visualização do parasita. O exame afresco do sedimento do lavado bucal revelou 100% de positividade e nítida visualização do parasita. Nenhuma técnica de coloração dos esfregaços se mostrou adequada, apresentando o núcleo freqüentemente mascarado pelos vacúolos fagocíticos. Em preparações coradas por azul de toluidina e na microscopia eletrônica de transmissão pode-se observar caracteres morfológicos típicos do protozoário.Entamoeba gingivalis is found only in its trophozoite form and it is postulated that its main transmission mechanism is through the kiss. E. gingivalis is considered pathogenic by some authors and commensal to others. It does not have a defined role in the installation of disease. To address some of this questions we studied a 100 patients who were seen through the Odontological Hospital from the Universidade Federal de Uberlândia in order to determine its frequency in the buccal cavity. The material were collected using swabs from four different buccal sites and the smears were stained by a modified Papanicolaou technique. The results revealed positivity index of 62%. The affinity of the dye to the food vacuole contents and to the ingested bactérias prevents clear visualisation of the central and peripherical chromatin constituents of the parasite's nucleus. Mouth washes with 3ml of saline from 10 patients, were used to evaluate which parasitological method of diagnosis (fresh, iron-haematoxylin stained, Giemsa and Papanicolaou gives better visualisation of the parasite. The mouth washes sediment from fresh material revealed 100% of positivity and clear visualisation of the free form and locomotion of the trophozoites. No stained technique of the smear showed adequate visualisation, presenting the nucleus partially covered by the food vacuoles. In stained preparations by toluidine blue ultrastructure analysis of the morphology of parasite can be obsewed.

Silvio Favoreto Junior

1995-12-01

178

[Halicephalobus gingivalis infection in a 5-year-old Tinker gelding].  

Science.gov (United States)

A 5-year old Tinker gelding was referred to the Department of Equine Sciences with a left eye uveitis and fever. At presentation the horse showed a mild lethargy, fever and decreased vision of the left eye. Rectal examination revealed an enlarged left kidney, with a hard and an irregular surface. The cranial mesentery artery had an enlarged and irregular aspect. Blood analysis showed anaemia, leucocytosis, increased blood urea nitrogen and creatinine and a hyperproteinemia. Urine analysis repeatedly showed a marked proteinuria and an increased gammaGT/creatinine ratio. The amount of abdominal fluid was slightly increased. However, the aspect, amount of cells and protein were normal. In the following two days the fever persisted and the horse showed anorexia and severe neurological signs. The horse was euthanized with permission of the owner. Post mortem examination showed a generalized parasitic infestation of Halicephalobus gingivalis in the uvea of the left eye, the kidneys and the central nerve system. PMID:16502977

Boswinkel, M; Neyens, I J S; Sloet van Oldruitenborgh-Oosterbaan, M M

2006-02-01

179

Porphyromonas gingivalis Fimbriae Inhibit Caspase-3-Mediated Apoptosis of Monocytic THP-1 Cells under Growth Factor Deprivation via Extracellular Signal-Regulated Kinase-Dependent Expression of p21 Cip/WAF1  

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Apoptotic regulation of monocytes/macrophages appears to be closely associated with chronic inflammatory reactions. Since it was demonstrated earlier that certain bacterial cell components are involved in apoptotic regulation of these cells, in the present study, we investigated whether the bacterial fimbria, an important cell structure involved in bacterial adherence to host cells, regulates apoptosis of human monocytic THP-1 cells induced under growth factor deprivation. To investigate this...

Ozaki, Ken; Hanazawa, Shigemasa

2001-01-01

180

Distinct lipid a moieties contribute to pathogen-induced site-specific vascular inflammation.  

Science.gov (United States)

Several successful pathogens have evolved mechanisms to evade host defense, resulting in the establishment of persistent and chronic infections. One such pathogen, Porphyromonas gingivalis, induces chronic low-grade inflammation associated with local inflammatory bone loss and systemic inflammation manifested as atherosclerosis. P. gingivalis expresses an atypical lipopolysaccharide (LPS) structure containing heterogeneous lipid A species, that exhibit Toll-like receptor-4 (TLR4) agonist or antagonist activity, or are non-activating at TLR4. In this study, we utilized a series of P. gingivalis lipid A mutants to demonstrate that antagonistic lipid A structures enable the pathogen to evade TLR4-mediated bactericidal activity in macrophages resulting in systemic inflammation. Production of antagonistic lipid A was associated with the induction of low levels of TLR4-dependent proinflammatory mediators, failed activation of the inflammasome and increased bacterial survival in macrophages. Oral infection of ApoE(-/-) mice with the P. gingivalis strain expressing antagonistic lipid A resulted in vascular inflammation, macrophage accumulation and atherosclerosis progression. In contrast, a P. gingivalis strain producing exclusively agonistic lipid A augmented levels of proinflammatory mediators and activated the inflammasome in a caspase-11-dependent manner, resulting in host cell lysis and decreased bacterial survival. ApoE(-/-) mice infected with this strain exhibited diminished vascular inflammation, macrophage accumulation, and atherosclerosis progression. Notably, the ability of P. gingivalis to induce local inflammatory bone loss was independent of lipid A expression, indicative of distinct mechanisms for induction of local versus systemic inflammation by this pathogen. Collectively, our results point to a pivotal role for activation of the non-canonical inflammasome in P. gingivalis infection and demonstrate that P. gingivalis evades immune detection at TLR4 facilitating chronic inflammation in the vasculature. These studies support the emerging concept that pathogen-mediated chronic inflammatory disorders result from specific pathogen-mediated evasion strategies resulting in low-grade chronic inflammation. PMID:25010102

Slocum, Connie; Coats, Stephen R; Hua, Ning; Kramer, Carolyn; Papadopoulos, George; Weinberg, Ellen O; Gudino, Cynthia V; Hamilton, James A; Darveau, Richard P; Genco, Caroline A

2014-07-01

 
 
 
 
181

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

Science.gov (United States)

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. PMID:23277124

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

2012-01-01

182

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

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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

2012-12-01

183

Lipopolysaccharide interaction with rabbit erythrocyte membranes.  

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In this study we have characterized the association of a polysaccharide-deficient, lipid-rich lipopolysaccharide (LPS) with rabbit erythrocytes (RaRBC). With polymyxin B sulfate-mediated hemolysis as a probe, we have shown that Salmonella minnesota R595 LPS interacts with RaRBC in two distinguishable steps. The first step whereby RaRBC exposed to LPS are rendered sensitive to polymyxin B-initiated lysis probably represents absorption of LPS to the RaRBC membrane. We investigated two possible ...

Carr, C.; Morrison, D. C.

1984-01-01

184

Biological characterization of Campylobacter fetus lipopolysaccharides.  

Science.gov (United States)

Lipopolysaccharides (LPS) of three strains of Campylobacter fetus (subspp. fetus and venerealis, and serotypes A and B), a bacterium of veterinary importance but also a cause of various infections in humans, were assessed for their ability to induce mitogenicity, gelation of Limulus amebocyte lysate, lethal toxicity in mice, and pyrogenicity in rabbits. All C. fetus LPS exhibited activities lower than those of Salmonella typhimurium LPS. LPS of C. fetus subsp. fetus serotype A had the lowest activity in all assays. Since the majority of C. fetus subsp. fetus isolates from humans are serotype A, the lower biological activities of this LPS may aid the pathogenesis of such strains. The lower activities of C. fetus LPS compared with those of S. typhimurium LPS may reflect the presence of longer fatty acid chains in the lipid A of C. fetus LPS, whereas interstrain differences in C. fetus LPS bioactivities may be related to some property influenced by composition of the saccharide moiety. PMID:8871115

Moran, A P; O'Malley, D T; Vuopio-Varkila, J; Varkila, K; Pyhälä, L; Saxén, H; Helander, I M

1996-08-01

185

Lipopolysaccharides of two strains of the phototrophic bacterium Rhodopseudomonas capsulata.  

Science.gov (United States)

The lipopolysaccharides of Rhodopseudomonas capsulata strains St. Louis (ATCC 23782) and Sp 11 both contain L-acofriose, rhamnose, glucose and glucosamine as the main sugar constituents. 2-Keto-3-deoxyoctonate and neuraminic acid were tentatively identified. The fatty acid spectrum found with both strains comprises 3-OH-C10 and C12:1 (ester-linked) and 3-oxo-C14 (amide-linked). Isolated lipid A from strain Sp 11 contains glucosamine, glucosamine-phosphate and the total of the fatty acids of the lipopolysaccharide. Methylation analysis of the degraded polysaccharide of this lipopolysaccharide shows L-acofriose in both terminal and 1 leads to 2 chain-linked positions in a 1:4 molar ratio. Rhamnose is exclusively chain-linked (1 leads to 2), glucose is both terminally and chain-linked (1 leads to 6) in a 1:1 molar ratio. The serological activity of the lipopolysaccharide of both the R. capsulata strains is low in antisera against living or heat-killed cells when tested by passive hemagglutination, Ouchterlony immunoprecipitation or gel-immunoelectrophoresis. No crossreaction was observed among the lipopolysaccharides of R. capsulata strains St. Louis, Sp 11 and 37b4 in immunoprecipitation. Lipopolysaccharide of strain Sp 11 was found to lack lethal toxicity in galactosamine-sensitized mice. PMID:6615126

Omar, A S; Flammann, H T; Borowiak, D; Weckesser, J

1983-06-01

186

Binding between lipopolysaccharide and cecropin A.  

Science.gov (United States)

Cecropin A (CA), a bioactive peptide, produced significant lethality to Pantoea agglomerans (PA) at low concentrations. Significant mortality occurred immediately after addition of CA. Separate preincubations of lipopolysaccharides (LPS) from the following bacteria: PA, Serratia marcescens, Escherichia coli (EC), and Salmonella typhimurium with CA were performed prior to the bioassay. CA was also preincubated with diphosphoryl lipid A (DPL-A) from EC and S. minnesota (SM), trilinolein, palmitic, lauric and myristic acids (fatty acids contained in the lipid A of PA-LPS) and bovine brain gangliosides. Spectral analysis to determine the interaction between glycosphingolipids (sphingomyelin, bovine brain gangliosides, and galactocerebrosides) and CA were performed. Results showed that all types of LPS and DPL-A as well as gangliosides studied blocked CA lethality to PA. The level of inhibition of CA antibacterial properties was dependent on LPS and DPL-A concentration. The individual fatty acids and trilinolein did not affect CA lethality to PA. Spectral studies showed complexation between CA and PA-LPS, both types of DPL-A, and the glycosphingolipids. Biological and chemical analyses confirm that CA binds to the diphosphoryl lipid A moiety of LPS. PMID:8569759

De Lucca, A J; Jacks, T J; Brogden, K A

1995-10-18

187

Lipopolysaccharide Extracellular Emulsifier Produced by Penicillium citrinum  

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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

2006-01-01

188

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

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English 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 tre [...] atment. Female rats received 100 ?g/kg lipopolysaccharide (LPS) or .9% saline solution on gestation day 18. Reproductive performance was observed at birth. On lactation days (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.

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

189

Curcumin attenuates lipopolysaccharide-induced renal inflammation.  

Science.gov (United States)

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. PMID:21415532

Zhong, Fang; Chen, Hui; Han, Lin; Jin, Yuanmeng; Wang, Weiming

2011-01-01

190

Fermentable non-starch polysaccharides increases the abundance of Bacteroides-Prevotella-Porphyromonas in ileal microbial community of growing pigs.  

Science.gov (United States)

Most plant-origin fiber sources used in pig production contains a mixture of soluble and insoluble non-starch polysaccharides (NSP). The knowledge about effects of these sources of NSP on the gut microbiota and its fermentation products is still scarce. The aim of this study was to investigate effects of feeding diets with native sources of NSP on the ileal and fecal microbial composition and the dietary impact on the concentration of short-chain fatty acids (SCFA) and lactic acid. The experiment comprised four diets and four periods in a change-over design with seven post valve t-cecum cannulated growing pigs. The four diets were balanced to be similar in NSP content and included one of four fiber sources, two diets were rich in pectins, through inclusion of chicory forage (CFO) and sugar beet pulp, and two were rich in arabinoxylan, through inclusion of wheat bran (WB) and grass meal. The gut microbial composition was assessed with terminal restriction fragment (TRF) length polymorphism and the abundance of Lactobacillus spp., Enterobacteriaceae, Bacteroides-Prevotella-Porphyromonas and the ?-xylosidase gene, xynB, were assessed with quantitative PCR. The gut microbiota did not cluster based on NSP structure (arabinoxylan or pectin) rather, the effect was to a high degree ingredient specific. In pigs fed diet CFO, three TRFs related to Prevotellaceae together consisted of more than 25% of the fecal microbiota, which is about 3 to 23 times higher (Pfiber (r=0.60; P=0.001) in ileal digesta were positively correlated with an increased abundance of Bacteroides-Prevotella-Porphyromonas. The effect on SCFA was correlated to specific neutral sugars where xylose increased the ileal butyric acid proportion, whereas arabinose increased the fecal butyric acid proportion. Moreover, chicory pectin increased the acetic acid proportion in both ileal digesta and feces. PMID:25046106

Ivarsson, E; Roos, S; Liu, H Y; Lindberg, J E

2014-11-01

191

DMPD: Lipopolysaccharide signaling in endothelial cells. [Dynamic Macrophage Pathway CSML Database  

Full Text Available 16357866 Lipopolysaccharide signaling in endothelial cells. Dauphinee SM, Karsan A. Lab Invest. ... signaling in endothelial cells. PubmedID 16357866 Title ... Lipopolysaccharide signaling in endothelial cells. ...

192

Augmented immunological activities of recombinant lipopolysaccharide possessing the mannose homopolymer as the O-specific polysaccharide.  

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Recombinant lipopolysaccharide possessing the mannose homopolymer as the O-specific polysaccharide was manufactured genetically by transforming Escherichia coli K-12 with various rfb genes capable of synthesizing the mannose homopolymer. Recombinant lipopolysaccharide exhibited levels of anticomplement activity, adjuvant activity, and regional lymph node-enlarging activity much higher than those exhibited by the original rough-type lipopolysaccharide from E. coli K-12 or lipopolysaccharide po...

Paeng, N.; Kido, N.; Schmidt, G.; Sugiyama, T.; Kato, Y.; Koide, N.; Yokochi, T.

1996-01-01

193

Bacterial lipopolysaccharide enhanced immunological responsiveness in exposed rats  

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Full Text Available n order to investigate the effects of the bacterial lipopolysaccharide (LPS on the brain and serum protein content, LPS was stereotaxically infused into the substantia nigra (SN of rats at different dosages (3 ?g, 10 ?g or 250 ?g/kg. The results showed that 7 days after neurosurgery there is no variations in brain protein content, while in serum protein content, the amount of gamma-globulins significantly increased compared to sham-operated control rats. The results suggested an immunologic response in the injected rats exposed to the bacterial lipopolysaccharide.

Lucian Gorgan

2009-09-01

194

Endotoxin-like activities of mycoplasmal lipopolysaccharides (lipoglycans).  

Science.gov (United States)

Lipoglycans (previously designated lipopolysaccharides) from several species of Acholeplasma and from Thermoplasma acidophilum were examined for endotoxin-like activities as measured by the standard rabbit fever test and the Limulus amoebocyte lysate assay. The lipoglycans from Acholeplasma granularum, Achloplasma laidlawii, Acholeplasma modicum, and Acholeplasma oculi caused a febrile response at concentrations of 1 ng/ml per kg or greater, whereas with control Escherichia coli EC-2 lipopolysaccharides, 6.25 ng/ml per kg was required. Similar results were obtained in the Limulus amoebocyte lysate test. The minimum concentrations in nanograms per milliliter required to stimulate formation of a solid clot were: Acholeplasma axanthum, 0.22; A. granularum, 0.85; A. modicum, 0.51; A. laidlawii, 1.05; A. oculi, 0.74. Standard E. coli 1B lipopolysaccharide required a concentration of 0.125 ng/ml. Thermoplasma lipoglycan was least active, requiring 4.25 ng/ml. Clotting of the Limulus lysate proceeds by the activation by lipopolysaccharide plus Ca(2+) of a proenzyme which cleaves an arginine-lysine peptide bond of the coagulogen. The clotting and amidase activities are inactivated by deoxycholate and can be reactivated by addition of lipopolysaccharide and Ca(2+). As with E. coli 1B lipopolysaccharide, acholeplasmal lipoglycans were shown to restore both clotting and amidase activities of the deoxycholate-inactivated Limulus clotting enzyme. The degree of restoration of amidase activity by mycoplasmal lipoglycans relative to E. coli lipopolysaccharide (1.00) were: A. axanthum, 1.71; A. modicum, 1.22; A. granularum, 0.61; and Thermoplasma, 0.37. The coagulating enzyme, restored with either E. coli lipopolysaccharide or mycoplasmal lipoglycans, was able to react with the synthetic peptide benzoyl-Ile-Glu-(gamma-OCH(3))-Gly-p-nitroaniline (an analog of the coagulogen) or with the purified coagulogen itself to form the clot. The mycoplasmal lipoglycans alone were incapable of promoting these reactions when incubated with the synthetic peptide or with the purified coagulogen, thereby ruling out the contamination of these lipoglycans with proteases capable of cleaving the same Arg-Lys peptide bond of the coagulogen. These results show that acholeplasmal lipoglycans possess endotoxin-like activities. Their passive or active role in disease remains to be established. PMID:7429642

Seid, R C; Smith, P F; Guevarra, G; Hochstein, H D; Barile, M F

1980-09-01

195

DMPD: Structural and functional analyses of bacterial lipopolysaccharides. [Dynamic Macrophage Pathway CSML Database  

Full Text Available 12106784 Structural and ... functional analyses of bacterial lipopolysaccharid es. Caroff M, Karibian ... D , Cavaillon JM, Haeffner-Cavaillon N. Microbes Infe ... 6. (.png) (.svg) (.html) (.csml) Show Structural and ... functional analyses of bacterial lipopolysaccharid ... es. Pubmed ID ... 12106784 Title Structural and ... functional analyse ... s of bacterial lipopolysaccharid es. Authors Caroff M, Karibian D , Cavaillon JM, Hae ...

196

Suppurative otitis and ascending meningoencephalitis associated with Bacteroides tectus and Porphyromonas gulae in a captive Parma wallaby (Macropus parma) with toxoplasmosis.  

Science.gov (United States)

A 6-year-old female Parma wallaby (Macropus parma) at a zoo in California developed acute ataxia and left-sided circling. Despite intensive care, clinical signs progressed to incoordination and prostration, and the animal was euthanized. At necropsy, the left tympanic cavity was filled with homogeneous suppurative exudate that extended into the cranium expanding the meninges and neuroparenchyma in the lateral and ventral aspect of the caudal ipsilateral brainstem and medulla oblongata. Microscopically, the brainstem showed regional severe suppurative meningoencephalitis with large numbers of neutrophils, fewer macrophages, and lymphocytes admixed with fibrin, necrotic cellular debris, hemorrhage, and mineralization, with numerous intralesional Gram-negative bacilli. Bacteroides spp. and Porphyromonas spp. were isolated on anaerobic culture from the meninges, and the bacteria were further characterized by partial 16S ribosomal RNA gene sequencing as Bacteroides tectus and Porphyromonas gulae. Bacterial aerobic culture from the meninges yielded very low numbers of mixed flora and Proteus spp., which were considered contaminants. Culture of Mycoplasma spp. from middle ear and meninges was negative. Additionally, Toxoplasma gondii cysts were detected by immunohistochemistry in the heart and brain, and anti-Toxoplasma antibodies were detected in serum. The genera Bacteroides and Porphyromonas have been associated with oral disease in marsupials; but not with otitis and meningoencephalitis. The results of the present work highlight the importance of performing anaerobic cultures in the diagnostic investigation of cases of suppurative otitis and meningoencephalitis in macropods. PMID:25057163

Giannitti, Federico; Schapira, Andrea; Anderson, Mark; Clothier, Kristin

2014-09-01

197

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. PMID:10049293

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

1999-01-01

198

Mechanisms of disposal of bacterial lipopolysaccharides by animal hosts.  

Science.gov (United States)

Much of the very extensive literature describing the (bio)chemistry and biology of bacterial lipopolysaccharides (LPS, endotoxin) has dealt with the properties of these molecules as potent triggers of host responses. This brief review will focus on what has been learned recently about mechanisms by which the host can dispose of LPS and counter its often excessive stimulatory effects. PMID:11008107

Elsbach, P

2000-08-01

199

Fatal equine meningoencephalitis in the United Kingdom caused by the panagrolaimid nematode Halicephalobus gingivalis: case report and review of the literature.  

Science.gov (United States)

A fatal case of eosinophilic and granulomatous meningoencephalitis caused by the free-living panagrolaimid nematode Halicephalobus gingivalis is reported in a 10-year-old Welsh gelding in the United Kingdom. Clinical examination first revealed behavioural abnormalities which rapidly progressed to severe ataxia, reduced mentation status and cranial nerve signs. Despite symptomatic treatment no amelioration of neurological signs was achieved and the horse was subjected to euthanasia. A complete post mortem examination revealed eosinophilic and granulomatous meningoencephalitis mainly affecting the cerebellum and brain stem with intralesional adult nematodes, larvae and eggs. There was also eosinophilic meningitis of the cervical spinal cord. The intralesional nematodes were morphologically consistent with the panagrolaimid nematode H. gingivalis. Although infection by this facultative neurotropic parasite is extremely rare, it needs to be considered in the differential diagnosis of central nervous signs in horses and, in particular, other equine helminthic infection of the central nervous system. This fatal case is unusual since lesions were locally very extensive and the nematodes did not colonise haematogenously to other organs as seen often in equine halicephalobosis. As the taxonomy of H. gingivalis has changed and some recent reports in the literature still refer to this species as Micronema deletrix or Halicephalobus deletrix, we here provide a short update of the species and some insights on the order Tylenchida, which contains free-living nematodes with parasitic tendencies. PMID:21496093

Hermosilla, C; Coumbe, K M; Habershon-Butcher, J; Schöniger, S

2011-11-01

200

Characterization of the Lipopolysaccharides and Capsules of Shewanella spp.  

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Electron microscopy, sodium dodecyl sulfate-polyacrylamide gel electrophoresis with silver staining and 1H, 13C, and 31P-nuclear magnetic resonance (NMR) were used to detect and characterize the lipopolysaccharides (LPSs) of several Shewanella species. Many expressed only rough LPS; however, approximately one-half produced smooth LPS (and/or capsular polysaccharides). Some LPSs were affected by growth temperature with increased chain length observed below 25°C. Maximum LPS heterogeneity was ...

Korenevsky, Anton A.; Vinogradov, Evgeny; Gorby, Yuri; Beveridge, Terry J.

2002-01-01

 
 
 
 
201

Innate immunity probed by lipopolysaccharides affinity strategy and proteomics.  

Science.gov (United States)

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. PMID:22752448

Giangrande, Chiara; Colarusso, Lucia; Lanzetta, Rosa; Molinaro, Antonio; Pucci, Piero; Amoresano, Angela

2013-01-01

202

Detectable serum flagellin and lipopolysaccharide and upregulated anti-flagellin and lipopolysaccharide immunoglobulins in human short bowel syndrome  

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Gut barrier dysfunction may occur in short bowel syndrome (SBS). We hypothesized that systemic exposure to flagellin and lipopolysaccharide (LPS) in SBS might regulate specific immune responses. We analyzed serial serum samples obtained from parenteral nutrition (PN)-dependent patients with SBS versus non-SBS control serum. Serum from 23 adult SBS patients was obtained at baseline and 4, 8, 12, 16, 20, and 24 wk in a trial of modified diet with or without growth hormone. Control serum was obt...

Ziegler, Thomas R.; Luo, Menghua; Esti?variz, Concepcio?n F.; Moore, Daniel A.; Sitaraman, Shanthi V.; Hao, Li; Bazargan, Niloofar; Klapproth, Jan-michael; Tian, Junqiang; Galloway, John R.; Leader, Lorraine M.; Jones, Dean P.; Gewirtz, Andrew T.

2008-01-01

203

Priming effect of fibronectin fragments on the macrophage inflammatory response: potential contribution to periodontitis.  

Science.gov (United States)

Fibronectin, an extracellular matrix component, is a substrate for multiple host and bacterial proteinases found in inflamed periodontal sites. In the present study, we investigated the potential contribution of various fibronectin fragments to the inflammatory process of periodontitis. Our results showed that the smaller fragments of fibronectin (30 and 45 kDa) were the most potent inflammatory inducers as they dose-dependently increased the secretion of TNF-?, IL-1?, and IL-8 by human macrophages. The 120-kDa fragment did not induce the secretion of all the cytokines tested, while intact fibronectin only increased IL-8 secretion and to a lesser extent TNF-? secretion. Cytokine secretion was associated with increased amounts of phosphorylated ERK1/2, JNK2, and p38? MAPK in treated macrophages. The combination of fibronectin or fibronectin fragments with Porphyromonas gingivalis lipopolysaccharide had an additive effect, but no synergism appeared to occur. It was also demonstrated that gingival crevicular fluid samples recovered from patients with moderate to severe periodontitis contained more fibronectin fragments than samples obtained from healthy subjects. Finally, both Arg- and Lys-gingipains purified from P. gingivalis were found to modulate fibronectin fragmentation. In conclusion, we showed that specific fibronectin fragments that may be present in diseased periodontal sites may contribute to maintaining and amplifying the inflammatory state and that P. gingivalis gingipains may be involved in the production of these fragments. PMID:22696147

Feghali, Karine; Grenier, Daniel

2012-10-01

204

Periodontal ligament and gingival fibroblasts participate in the production of TGF-?, interleukin (IL-8 and IL-10  

Directory of Open Access Journals (Sweden)

Full Text Available The aim of this study was to quantify and compare the production of transforming growth factor beta (TGF-?, interleukin (IL-8 and IL-10 by human cultured periodontal ligament and gingival fibroblasts both obtained from the same donors challenged with lipopolysaccharide (LPS from Porphyromonas gingivalis. Fibroblasts were exposed to 0.1-10 µg/mL of LPS from P. gingivalis and after 24 h the supernatants were collected and analyzed by enzyme-linked immunosorbent assay (ELISA. TGF-? protein production was upregulated in a concentration-dependent manner, mainly in gingival fibroblasts, which was statistically significant when challenged by 10 µg/mL LPS. Additionally, at this concentration, gingival fibroblasts had almost a two-fold increase in the amount of TGF-? when compared to periodontal ligament fibroblasts. Both periodontal ligament and gingival fibroblasts showed an increase in IL-8 production when challenged with 1 µg/mL and 10 µg/mL LPS. IL-10 production remained unaffected when challenged by any of the LPS concentrations tested in either periodontal ligament or gingival fibroblasts. Our results demonstrate that periodontal ligament and gingival fibroblasts when challenged by LPS from P. gingivalis with 24 h may play a critical role in producing TGF-? and IL-8 but not IL-10.

Ana Carolina de Faria Morandini

2011-04-01

205

Self-heat shock protein 60 induces tumour necrosis factor-alpha in monocyte-derived macrophage: possible role in chronic inflammatory periodontal disease.  

Science.gov (United States)

Heat shock protein 60 (hsp60) has been increasingly recognized as an important molecule in infectious and autoimmune diseases. We have demonstrated previously that serum antibodies to both human hsp60 and Porphyromonas gingivalis GroEL were elevated in periodontitis patients compared with healthy subjects. In order to clarify the relative importance of hsp60 in the inflammatory response in periodontal disease, the stimulatory effect of human and bacterial hsp60 on the production of tumour necrosis factor-alpha (TNF-alpha) was examined in phorbol myristate acetate (PMA)-stimulated THP-1 cells. As bacterial hsp60s, recombinant P. gingivalis and Actinobacillus actinomycetemcomitans GroEL was used. Human hsp60 but not P. gingivalis or A. actinomycetemcomitans GroEL demonstrated stimulatory activity similar to lipopolysaccharide (LPS) derived from the bacteria. The activity of hsp60 was inhibited by anti-CD14 and anti-Toll-like receptor 4 (TLR4) antibodies, suggesting that both CD14 and TLR4 mediate hsp60 signalling. Immunohistochemical analysis demonstrated that hsp60 is abundantly expressed in periodontitis lesions. Therefore, it is postulated that periodontopathic bacteria stimulate the cells in the periodontium to up-regulate the expression of hsp60, which in turn may stimulate macrophage and possibly other cells to produce proinflammatory cytokines. These mechanisms may be involved in the chronicity and tissue destruction of periodontal disease. PMID:11882035

Ueki, K; Tabeta, K; Yoshie, H; Yamazaki, K

2002-01-01

206

Lipopolysaccharide as a target for brucellosis vaccine design.  

Science.gov (United States)

The gram-negative bacteria of the genus Brucella are facultative intracellular parasites that cause brucellosis, a world wide-distributed zoonotic disease that represents a serious problem for animal and human health. There is no human-to-human contagion and, since there is no human vaccine, animal vaccination is essential to control brucellosis. However, current vaccines (all developed empirically) do not provide 100% protection and are infectious in humans. Attempts to generate new vaccines by obtaining mutants lacking the lipopolysaccharide O-polysaccharide, in purine metabolism or in Brucella type IV secretion system have not been successful. Here we propose a new approach to develop brucellosis vaccines based on the concept that Brucella surface molecules evade efficient detection by innate immunity, thus delaying protective Th1 responses and opening a time window to reach sheltered intracellular compartments. We showed recently that a branch of the core oligosaccharide section of Brucella lipopolysaccharide hampers recognition by TLR4-MD2. Mutation of glycosyltransferase WadC, involved in the synthesis of this branch, results in a lipopolysaccharide that, while keeping the O-polysaccharide essential for optimal protection, shows a truncated core, is more efficiently recognized by MD2 and triggers an increased cytokine response. In keeping with this, the wadC mutant is attenuated in dendritic cells and mice. In the mouse model of brucellosis vaccines, the Brucella abortus wadC mutant conferred protection similar to that provided by S19, the best cattle vaccine available. The properties of the wadC mutant provide the proof of concept for this new approach and open the way for more effective brucellosis vaccines. PMID:23219811

Conde-Álvarez, Raquel; Arce-Gorvel, Vilma; Gil-Ramírez, Yolanda; Iriarte, Maite; Grilló, María-Jesús; Gorvel, Jean Pierre; Moriyón, Ignacio

2013-05-01

207

Human monocyte CD14 is upregulated by lipopolysaccharide.  

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Membrane CD14 is involved in lipopolysaccharide (LPS)-induced monocyte activation; it binds LPS, and antibodies against CD14 block the effects of low-dose LPS. It is unknown how LPS regulates its own receptor CD14 in vitro. Therefore, we investigated the effects of LPS on CD14 mRNA and membrane and soluble CD14 (mCD14 and sCD14, respectively) in human monocytes and macrophages. No changes were observed during the first 3 h of LPS stimulation. After 6 to 15 h, LPS weakly reduced CD14 mRNA and ...

Landmann, R.; Knopf, H. P.; Link, S.; Sansano, S.; Schumann, R.; Zimmerli, W.

1996-01-01

208

Comparative immunochemistry of lipopolysaccharides from Branhamella catarrhalis strains.  

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Lipopolysaccharides (LPS) were extracted and purified from the type strain and from a clinical isolate of Branhamella catarrhalis. Chemical analysis revealed the presence of glucose, galactose, and glucosamine in different molar proportions in the LPS from these two isolates, whereas there was no difference between the two isolates in the ratios of ketodeoxyoctonate, phosphate, and the fatty acids C12, 3-OH-C12, and 3-OH-C11 present. Heptose or 3-OH-C14 was not detectable in either preparatio...

Fomsgaard, J. S.; Fomsgaard, A.; Høiby, N.; Bruun, B.; Galanos, C.

1991-01-01

209

In vitro study of 980nm diode laser in dental implant disinfection  

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Objective: To evaluate the potential of 980nm diode laser to reduce bacteria after irradiation of three different dental implant surfaces contaminated with Enterococcus faecalis and Porphyromonas gingivalis, as well as the possible changes in the irradiated implant surfaces.Methods: Seventy two implants with machined surfaces, airborne particle abraded with titanium oxide and acid-etched surfaces were exposed to Enterococcus faecalis and Porphyromonas gingivalis cultures and irradiated with 9...

Fábio Gonçalves; Artêmio Luiz Zanetti; Raquel Virgínia Zanetti; Saturnino Aparecido Ramalho

2009-01-01

210

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

2012-10-01

211

DMPD: Signal transduction by the lipopolysaccharide receptor, Toll-like receptor-4. [Dynamic Macrophage Pathway CSML Database  

Full Text Available 15379975 Signal ... transduction by the lipopolysaccharide receptor, Toll-like receptor-4. Palsson-M ... ;113(2):153-62. (.png) (.svg) (.html) (.csml) Show Signal ... transduction by the lipopolysaccharide receptor, T ... oll-like receptor-4. PubmedID 15379975 Title Signal ... transduction by the lipopolysaccharide receptor, T ...

212

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

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english 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 morpholo [...] gies 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

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

1027-10-01

213

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

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english 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 morpholo [...] gies 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

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

214

A method for unobtrusive labeling of lipopolysaccharides with quantum dots.  

Science.gov (United States)

Bacterial endotoxins or lipopolysaccharides (LPS) are among the most potent activators of innate immune system, yet mechanisms of their action and, in particular, the role of the glycans remain elusive. Efficient noninvasive labeling strategies are necessary for studying interactions of LPS glycans with biological systems. Here, we describe a new method for labeling LPS and other lipoglycans with luminescent quantum dots (QDots). The labeling is achieved by the partitioning of hydrophobic quantum dots into the core of various LPS aggregates without disturbing the native LPS structure. The biofunctionality of the LPS-QDot conjugates is demonstrated by labeling of mouse monocytes. This simple method will find broad applicability in studies concerned with visualization of LPS biodistribution and identification of LPS-binding agents. PMID:21567322

Morales-Betanzos, Carlos; Gonzalez-Moa, Maria; Svarovsky, Sergei A

2011-01-01

215

Antioxidant effects of cranberry powder in lipopolysaccharide treated hypercholesterolemic rats.  

Science.gov (United States)

This study was conducted to investigate the effects of cranberry power on antioxidant defense system in rats fed an atherogenic diet and injected with lipopolysaccharide (LPS). Sprague-Dawley rats were divided into the following 5 groups: normal diet+saline (NS), atherogenic diet+saline (AS), atherogenic diet+LPS (AL), atherogenic diet with 5% cranberry powder+LPS (AL-C5), and atherogenic diet with 10% cranberry powder+LPS (AL-C10). Total antioxidant status measured by ferric reducing ability of plasma (FRAP) was significantly reduced by LPS injection (24%) and was restored by the cranberry powder treatment (Pcranberry powder was incorporated in to the diet. Activity of serum superoxide dismutase (SOD) tended to be lowered by LPS injection and declined further in cranberry powder fortified groups. Overall results indicate that dietary cranberry powder may provide appropriate antioxidants to counter the diminished antioxidant status induced by exposing hypercholesterolemic rats to LPS. PMID:25054105

Kim, Mi Joung; Kim, Jung Hee; Kwak, Ho-Kyung

2014-06-01

216

Pleurotus eryngii Ameliorates Lipopolysaccharide-Induced Lung Inflammation in Mice.  

Science.gov (United States)

Pleurotus eryngii (P. eryngii) is consumed as a fresh cultivated mushroom worldwide and demonstrated to have multiple beneficial effects. We investigated the anti-inflammatory effect of P. eryngii in mice with acute lung injury (ALI). Intranasal instillation of lipopolysaccharide (LPS) (10? ? g/site/mouse) induced marked lung inflammation (increase in the number of inflammatory cells, protein leakage, and production of nitric oxide in bronchoalveolar lavage fluid) as well as histopathological damage in the lung, 6?h after treatment. Mice administered heat-treated P. eryngii (0.3-1?g/kg, p.o. (HTPE)) 1?h before LPS challenge showed decreased pulmonary inflammation and ameliorated histopathological damage. These results suggest that HTPE has anti-inflammatory effects against ALI. Thus, P. eryngii itself may also have anti-inflammatory effects and could be a beneficial food for the prevention of ALI induced by bacterial infection. PMID:24799939

Kawai, Junya; Andoh, Tsugunobu; Ouchi, Kenji; Inatomi, Satoshi

2014-01-01

217

[Immunostimulating activity of the lipopolysaccharides of blue-green algae].  

Science.gov (United States)

The whole cells of blue-gree algae and lipopolysaccharides isolated from these cells were shown to stimulate the production of macro-(mainly) and microglobulin antibodies in rabbits. The macro- and microphage indices in rabbits increased significantly after the injection of LPS isolated from blue-green algae 24--48 hours before infecting the animals with a virulent Y. pseudotuberculosis strain. Besides, the inhibiting action of this strain on the migration of phagocytes to the site of infection was abolished immediately after the injection. The use of the indirect hemagglutination test allowed to prove the absence of close antigenic interrelations between blue-green algae and the following organisms: Spirulina platensis, Microcystis aeruginosa, Phormidium africanum and P. uncinatum. PMID:117655

Besednova, N N; Smolina, T P; Mikhe?skaia, L V; Ovodova, R G

1979-12-01

218

Drugs composed of polysaccharides obtained from lipopolysaccharides extracted from bacteria  

International Nuclear Information System (INIS)

This invention concerns a protection and control agent against irradiation in living beings, composed of polysaccharides, called PS, obtained from lipopolysaccharides (LPS), which in turn are extracted from Gram-negative bacteria after separation of the major part and preferably all the initial lipid content of these LPS's. The molecular weight of the PS ingredients, once purified, is between 8,000 and 35,000 and mainly around 10,000. They contain: around 50 to 70% carbon hydrates by weight; around 1 to 8% hexoamines by weight; around 1 to 5% amino-compounds, among which 1 to 2% amino-acids by weight less than 1% and preferably no fatty acids or lipids and their phosphorus content is around 0.2 to 0.5% by weight

219

Effects of Salmonella typhimurium lipopolysaccharide on broiler chickens.  

Science.gov (United States)

The effects of Salmonella typhimurium lipopolysaccharide (LPS) on the physiology of 3-wk-old broiler chickens were studied at 12, 24, and 48 h after a single intravenous injection of saline or LPS. Lipopolysaccharide elevated cloacal temperature by 3 h after injection, induced a diuretic response, and decreased BW gain. An increase in the relative liver weight was evident in LPS-treated birds at all time intervals, whereas a decrease in the relative weight of bursa of Fabricius was observed only at the 48-h time point. The plasma interleukin (IL)-6 and the blood heterophil concentrations were elevated at 12 and 24 h following LPS administration. These changes were not observed in control chickens or in LPS-treated chickens at 48 h. A decrease in the blood glucose concentration in LPS-treated birds at 12 h was accompanied by an elevation in the blood phosphate level. An increase in total plasma protein concentration was observed only at 24 and 48 h after LPS treatment. Comparative SDS-PAGE analysis of plasma proteins from these birds under nonreducing conditions showed some quantitative differences in four bands of proteins between saline and LPS-treated chickens. A protein corresponding to an approximate molecular weight (MW) of 65 kDa increased in LPS-treated chickens, and three other proteins with MW of approximately 39, 49, and 56 kDa showed reductions in concentration compared with saline-treated controls. These results show that LPS induces a number of physiological changes that may be responsible for the regulation of the acute phase response in chickens. PMID:10685886

Xie, H; Rath, N C; Huff, G R; Huff, W E; Balog, J M

2000-01-01

220

Rapid presumptive identification of black-pigmented gram-negative anaerobic bacteria by using 4-methylumbelliferone derivatives.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

A rapid method for presumptive identification of black-pigmented gram-negative anaerobic rods was developed. Using filter paper spot tests for indole production, sialidase, alpha-glucosidase, beta-glucosidase, alpha-fucosidase, and trypsinlike enzyme activities, 100% of Porphyromonas gingivalis, Prevotella intermedia, and Bacteroides levii and 89% of Prevotella corporis isolates were correctly identified to the species level. Porphyromonas asaccharolytica and Porphyromonas endodontalis could ...

Moncla, B. J.; Braham, P.; Rabe, L. K.; Hillier, S. L.

1991-01-01

 
 
 
 
221

Pathogen-induced inflammation at sites distant from oral infection: bacterial persistence and induction of cell-specific innate immune inflammatory pathways  

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A hallmark of infection with the gram-negative pathogen Porphyromonas gingivalis is the induction of a chronic inflammatory response. P. gingivalis induces a local chronic inflammatory response that results in oral inflammatory bone destruction, which manifests as periodontal disease. In addition to chronic inflammation at the initial site of infection, mounting evidence has accumulated supporting a role for P. gingivalis-mediated periodontal disease as a risk factor for several systemic dise...

Hayashi, C.; Gudino, C. V.; Gibson, F. C.; Genco, C. A.

2010-01-01

222

Pathogen-Accelerated Atherosclerosis Occurs Early after Exposure and Can Be Prevented via Immunization  

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Here we report on early inflammatory events associated with Porphyromonas gingivalis-accelerated atherosclerosis in apolipoprotein E knockout (ApoE?/?) mice. Animals challenged with P. gingivalis presented with increased macrophage infiltration, innate immune marker expression, and atheroma without elevated systemic inflammatory mediators. This early local inflammatory response was prevented in mice immunized with P. gingivalis. We conclude that localized up-regulation of innate immune ma...

Miyamoto, Takanari; Yumoto, Hiromichi; Takahashi, Yusuke; Davey, Michael; Gibson, Frank C.; Genco, Caroline Attardo

2006-01-01

223

Entamoeba gingivalis y Trichomonas tenax en cavidad bucal de pacientes de la Clínica Integral del Adulto de la Facultad de Odontología, Maracaibo, Venezuela / Entamoeba gingivalis and tricomonas tenax in the oral cavity of patients from the integral adult clinic of the faculty of odontology, Maracaibo, Venezuela  

Scientific Electronic Library Online (English)

Full Text Available SciELO Venezuela | Language: Spanish Abstract in spanish Para determinar la prevalencia de Entamoeba gingivalis y Trichomonas tenax en cavidad bucal, se analizaron 50 muestras de la cavidad bucal de individuos de ambos géneros que acudieron a la Clínica Integral del Adulto de la Facultad de Odontología de la Universidad del Zulia. Se dividieron en dos gru [...] pos, de 25 individuos cada uno. Grupo 1, con manifestaciones clínicas de enfermedad (enfermedad periodontal y/o caries dental) al cual se le tomaron muestras de caries dental, placa y cálculo dental y grupo 2 o control con cavidad bucal sin manifestaciones clínicas de enfermedad, al cual se le tomó muestras de saliva y placa dental. Las muestras fueron analizadas microscópicamente a través del examen directo y con coloración permanente de hematoxilina férrica. Se observó una prevalencia de protozoarios bucales de un 10%; la especie predominante fue Entamoeba gingivalis en 5 casos, seguida de Trichomonas tenax en 1 caso. El estrato de 20 a 39 años fue el más afectado con un 10% de los casos. Al realizar el análisis estadístico resultó significativo (p=0,011) para las variables parasitismo y cavidad bucal enferma. El presente estudio pone de manifiesto una baja prevalencia de los protozoarios bucales en la población estudiada. Abstract in english Fifty samples from the oral cavity of individuals of both genders who attended the Integral Adult Clinic of the Faculty of Odontology of Universidad del Zulia were analyzed to determine Entamoeba gingivalis and Trichomonas tenax prevalence. The patients were divided into two groups of 25 individuals [...] each: Group 1, with clinical disease manifestations (periodontal disease and/or dental caries) from which we took samples from dental caries, plaque and dental calculus; and Group 2 or control, who had no clinical disease manifestations, from which we took saliva and dental plaque samples. All samples were analyzed microscopically through direct examination and with a ferric hematoxilin stain. There was a 10% prevalence of oral protozoa; the predominant species was Entamoeba gingivalis in 5 cases followed by Trichomonas tenax in 1 case. The 20-39 years age group was the most affected with 10% of cases. The statistical analysis was significant (p=0.011) for the parasitism and diseased oral cavity variables. The present study shows a low prevalence of oral cavity protozoa in the population studied.

Ellen Mabel, Acurero Osorio; Adriana Beatriz, Maldonado Ibáñez; Carla Maldonado, Ibáñez; Angela María, Bracho Mora; Jennifer, Parra; Yennifer, Urdaneta; Maryorie, Urdaneta.

2009-12-01

224

Cloning and characterization of a gene from Pasteurella haemolytica A1 involved in lipopolysaccharide biosynthesis.  

Science.gov (United States)

A Pasteurella haemolytica A1 gene involved in the biosynthesis of a moiety on the core of the lipopolysaccharide molecule has been cloned and characterized. Escherichia coli clones which carry this gene showed an alteration of its lipopolysaccharide migration profile on tricine SDS-PAGE and exhibited resistance to the core-specific phage U3. In addition, lipopolysaccharide extracted from the E. coli clones was recognized by an anti-corespecific antiserum, but not by antiserum specific for the O antigen of P. haemolytica A1 lipopolysaccharide. Nucleotide sequence analysis of the cloned DNA identified an open reading frame (lpsA) coding for a protein of 263 amino acids which showed significant homology with a Haemophilus influenzae type b lipooligosaccharide biosynthesis gene. PCR amplification of genomic DNA, using primers based on the P. haemolytica A1 lpsA sequence, yielded products from only the A biotypes of P. haemolytica. PMID:7781993

Potter, M D; Lo, R Y

1995-06-01

225

Proteomics of Fusobacterium nucleatum within a model developing oral microbial community.  

Science.gov (United States)

Fusobacterium nucleatum is a common oral organism that can provide adhesive and metabolic support to developing periodontal bacterial communities. It is within the context of these communities that disease occurs. We have previously reported whole cell proteomics analyses of Porphyromonas gingivalis and Streptococcus gordonii in early-stage communities with each other and with F. nucleatum, modeled using 18 h pellets. Here, we report the adaptation of F. nucleatum to the same experimental conditions as measured by differential protein expression. About 1210 F. nucleatum proteins were detected in single species F. nucleatum control samples, 1192 in communities with P. gingivalis, 1224 with S. gordonii, and 1135 with all three species. Quantitative comparisons among the proteomes revealed important changes in all mixed samples with distinct responses to P. gingivalis or S. gordonii alone and in combination. The results were inspected manually and an ontology analysis conducted using DAVID (Database for annotation, visualization, and integrated discovery). Extensive changes were detected in energy metabolism. All multispecies comparisons showed reductions in amino acid fermentation and a shift toward butanoate as a metabolic byproduct, although the two organism model community with S. gordonii showed increases in alanine, threonine, methionine, and cysteine pathways, and in the three species samples there were increases in lysine and methionine. The communities with P. gingivalis or all three organisms showed reduced glycolysis proteins, but F. nucleatum paired with S. gordonii displayed increased glycolysis/gluconeogenesis proteins. The S. gordonii containing two organism model also showed increases in the ethanolamine pathway while the three species sample showed decreases relative to the F. nucleatum single organism control. All of the nascent model communities displayed reduced translation, lipopolysaccharide, and cell wall biosynthesis, DNA replication and DNA repair. PMID:25155235

Hendrickson, Erik L; Wang, Tiansong; Beck, David A C; Dickinson, Brittany C; Wright, Christopher J; J Lamont, Richard; Hackett, Murray

2014-10-01

226

Hot-symbiont interactions. III. Purification and partial characterization of Rhizobium lipopolysaccharides  

Energy Technology Data Exchange (ETDEWEB)

The lipopolysaccharides of three strains each of Rhizobium leguminosarum, R. phasecoli, and R. trifolii have been purified and partially characterized. The last step in the purification procedure is gel filtration column chromatography using Sepharose 4B with the elution buffer consisting of ethylenediaminetetraacetic acid and triethylamine. Each of the lipopolysaccharides reported in this paper elutes as a symmetrical peak in the partially included volume of this Sepharose 4B column. The ration of 2-keto-3-deoxyoctonate acid (a sugar which is characteristic of lipopolysaccharides) to hexose is constant throughout the carbohydrate-containing peaks as they elute from the Sepharose 4B. The compositions and immunodominant structures of the purified lipopolysaccharides vary as much among strains of a single Rhizobium species as among the different species of Rhizobium. There is no obvious correlation between the nodulation group to which a Rhizobium belongs and the chemical composition or immunochemistry of the Rhizobium's lipopolysaccharide. There is extensive crosslysis by phage of strains of R. trifolii, R. phaseoli, and R. leguminosarum. This suggests that the receptors for these cross-lysing phage reside either in nonlipopolysaccharide structures or in common structures within the lipopolysaccharide which are not detected by compositional or immunochemical analysis.

Carlson, R.W.; Sanders, R.E.; Napoli, C.; Albersheim, P.

1978-01-01

227

Allergen immunotherapy with nanoparticles containing lipopolysaccharide from Brucella ovis.  

Science.gov (United States)

The adjuvant and protective capacity against anaphylactic shock of the association between rough lipopolysaccharide of Brucella ovis (LPS) coencapsulated with ovalbumin (OVA), as a model allergen, in Gantrez AN nanoparticles was investigated. Several strategies were performed in order to study the adjuvant effect of the LPS either encapsulated or coating the nanoparticles. OVA, as well as LPS, was incorporated either during the manufacturing process (OVA-encapsulated or LPS-encapsulated nanoparticles, respectively) or after the preparation (OVA-coated or LPS-coated nanoparticles, respectively). After the administration of 10 microg of OVA incorporated in the different formulations, all the nanoparticles, with or without LPS, were capable of amplifying the immune response (IgG(1) and IgG(2a)). However, in a model of sensitized mice to OVA, the formulation with OVA and LPS-entrapped inside the nanoparticles administered intradermally in three doses of 3 microg of OVA each was the only treatment that totally protected the mice from death after a challenge with an intraperitoneal injection of OVA. In contrast, the control group administered with OVA adsorbed onto a commercial alhydrogel adjuvant showed 80% mortality. These results are highly suggestive for the valuable use of Gantrez nanoparticles combined with rough LPS of B. ovis in immunotherapy. PMID:18582571

Gómez, Sara; Gamazo, Carlos; San Roman, Beatriz; Ferrer, Marta; Sanz, Maria Luisa; Espuelas, Socorro; Irache, Juan M

2008-11-01

228

Electrophoretic and immunoblot analysis of Campylobacter fetus lipopolysaccharides.  

Science.gov (United States)

Proteinase K-digested cell lysates from 25 Campylobacter fetus subspecies fetus and C. fetus subsp. venerealis strains were examined by SDS-PAGE and immunoblotting. Three SDS-PAGE lipopolysaccharide (LPS) profiles were observed. Two profiles were consistent with those previously reported for serogroup A and serogroup B and AB isolates and were distinguished by the relative mobility of bands in the O-chain region and by a strong reaction on immunoblots with homologous antisera. The third profile was similar but had faster migrating O-chain bands. Immunoblot reactions using homologous and heterologous adsorbed antisera showed that the O-antigen of the C. fetus subsp. fetus reference strain and other profile 2-type LPS strains was distinct from the O-antigens of strains with profile 1- or profile 3-type LPS. O-antigens of strains with profile 1- and profile 3-type LPS had shared epitopes. One strain had core components but no detectable O-antigens. Common core LPS antigens appear to be present in all strains and antibodies to common core LPS epitopes may be useful reagents for rapid detection of C. fetus. PMID:8828127

Brooks, B W; Garcia, M M; Robertson, R H; Lior, H

1996-07-01

229

Lipopolysaccharide neutralization by a novel peptide derived from phosvitin.  

Science.gov (United States)

Lipopolysaccharide (LPS), also known as endotoxin, is the primary trigger of sepsis, which is associated with high mortality in patients. No therapeutic agents are currently efficacious enough to protect patients from sepsis characterized by LPS-mediated tissue damage and organ failure. Previously, a phosvitin-derived peptide, Pt5, which consists of the C-terminal 55 residues of zebrafish phosvitin, has been shown to function as an antibacterial agent. In this study, we have generated six mutants by site-directed mutagenesis based on the sequence of Pt5, and found that one of the six mutants, Pt5e, showed the strongest bactericidal activities against Escherichia coli and Staphylococcus aureus. We then demonstrated that Pt5e was able to bind to LPS and lipoteichoic acid (LTA). More importantly, we showed that Pt5e significantly inhibited LPS-induced tumor-necrosis factor (TNF)-? and interleukin (IL)-1? release from murine RAW264.7 cells and considerably reduced serum TNF-? and IL-1? levels in mice. Additionally, Pt5e protected the liver from damage by LPS, and remarkably promoted the survival rate of the endotoxemia mice. Furthermore, Pt5e displayed no cytotoxicity to murine RAW264.7 macrophages and no hemolytic activity toward human red blood cells. These data together indicate that Pt5e is an endotoxin-neutralizing agent with a therapeutic potential in clinical treatment of LPS-induced sepsis. PMID:24028820

Hu, Lili; Sun, Chen; Wang, Shengnan; Su, Feng; Zhang, Shicui

2013-11-01

230

Lipopolysaccharide induced inflammation in the perivascular space in lungs  

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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.

Pabst Reinhard

2008-07-01

231

Detection of an Actinobacillus pleuropneumoniae serotype 2 lipopolysaccharide (LPS) variant  

DEFF Research Database (Denmark)

Until now 12 serotypes of Actinobacillus pleuropneumoniae have been recognized. The specificity of the serotypes reside in the carbohydrate composition of the capsular polysaccharides and lipopolysaccharides (LPS). The LPS of A. pleuropneumoniae serotype 2 is a smooth type LPS with O-chains of linear repeating pentasaccharide units with an O-acetyl group linked to a glucose unit. A monoclonal antibody (MAb 102-G02) directed against A. pleuropneumoniae serotype 2 was characterized in enzyme linked immunosorbent assay (ELISA) and in sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The MAI, 102-G02 was directed against an epitope on the O-chain of the LPS and was used to define a new LPS variant of A. pleuropneumoniae serotype 2 (referred to as A. pleuropneumoniae serotype 2X). Investigation of the reactivity of the MAb 102-G02 against an A. pleuropneumoniae serotype 2X field isolate (9008) and the Danish App-2 strain 4226 in electron microscopy, confirmed the different binding patterns.

1996-01-01

232

Passive transfer of leishmania lipopolysaccharide confers parasite survival in macrophages  

International Nuclear Information System (INIS)

Infection of macrophages by the intracellular protozoan parasite Leishmania involves specific attachment to the host membrane, followed by phagocytosis and intracellular survival and growth. Two parasite molecules have been implicated in the attachment event: Leishmania lipopolysaccharide (L-LPS) and a glycoprotein (gp63). This study was designed to clarify the role of L-LPS in infection and the stage in the process of infection at which it operates. The authors have recently identified a Leishmania major strain (LRC-L119) which lacks the L-LPS molecule and is not infective for hamsters or mice. This parasite was isolated from a gerbil in Kenya and was identified phenotypically as L. major by isoenzyme and fatty acid analysis. In this study they have confirmed at the genotype level that LRC-L119 is L. major by analyzing and comparing the organization of cloned DNA sequences in the genome of different strains of L. major. Here they show that LRC-L119 promastigotes are phagocytosed rapidly by macrophages in vitro, but in contrast to virulent strains of L. major, they are then killed over a period of 18 hr. In addition, they show that transfer of purified L-LPS from a virulent clone of L. major (V121) into LRC-L119 promastigotes confers on them the ability to survive in macrophages in vitro

233

Genomic and proteomic studies on Plesiomonas shigelloides lipopolysaccharide core biosynthesis.  

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We report here the identification of waa clusters with the genes required for the biosynthesis of the core lipopolysaccharides (LPS) of two Plesiomonas shigelloides strains. Both P. shigelloides waa clusters shared all of the genes besides the ones flanking waaL. In both strains, all of the genes were found in the waa gene cluster, although one common core biosynthetic gene (wapG) was found in a different chromosome location outside the cluster. Since P. shigelloides and Klebsiella pneumoniae share a core LPS carbohydrate backbone extending up at least to the second outer-core residue, the functions of the common P. shigelloides genes were elucidated by genetic complementation studies using well-defined K. pneumoniae mutants. The function of strain-specific inner- or outer-core genes was identified by using as a surrogate acceptor LPS from three well-defined K. pneumoniae core LPS mutants. Using this strategy, we were able to assign a proteomic function to all of the P. shigelloides waa genes identified in the two strains encoding six new glycosyltransferases (WapA, -B, -C, -D, -F, and -G). P. shigelloides demonstrated an important variety of core LPS structures, despite being a single species of the genus, as well as high homologous recombination in housekeeping genes. PMID:24244003

Aquilini, Eleonora; Merino, Susana; Regué, Miguel; Tomás, Juan M

2014-02-01

234

Visualization and analysis of lipopolysaccharide distribution in binary phospholipid bilayers  

International Nuclear Information System (INIS)

Lipopolysaccharide (LPS) is an endotoxin released from the outer membrane of Gram-negative bacteria during infections. It have been reported that LPS may play a role in the outer membrane of bacteria similar to that of cholesterol in eukaryotic plasma membranes. In this article we compare the effect of introducing LPS or cholesterol in liposomes made of dipalmitoylphosphatidylcholine/dioleoylphosphatidylcholine on the solubilization process by Triton X-100. The results show that liposomes containing LPS or cholesterol are more resistant to solubilization by Triton X-100 than the binary phospholipid mixtures at 4 oC. The LPS distribution was analyzed on GUVs of DPPC:DOPC using FITC-LPS. Solid and liquid-crystalline domains were visualized labeling the GUVs with LAURDAN and GP images were acquired using a two-photon microscope. The images show a selective distribution of LPS in gel domains. Our results support the hypothesis that LPS could aggregate and concentrate selectively in biological membranes providing a mechanism to bring together several components of the LPS-sensing machinery.

235

Lipopolysaccharide exposure augments isoniazide-induced liver injury.  

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Isoniazide (INH) is a classic antituberculosis drug associated with clinical idiosyncratic drug-induced liver injury. It has been hypothesized that the interaction between a drug and modest inflammation results in a decreased threshold for drug toxicity. In this study, we tested the hypothesis that INH causes liver injury in rats when coadministered with lipopolysaccharide (LPS). Neither INH nor LPS alone caused liver injury. The coadministration of INH and LPS was associated with increases in serum and histopathological markers of liver injury. Tumour necrosis factor-? expression was significantly increased in the coadministered group. The downregulation of the bile acid transporter, bile salt export pump, and multidrug resistance protein 2 at both mRNA and protein levels was observed. Furthermore, the level of Farnesoid X receptor, which regulates the bile salt export pump and multidrug resistance protein 2, were clearly decreased. These results indicate that the coadministration of nontoxic doses of LPS and INH causes liver injury; the disruption of biliary excretion is considered the primary inflammation-related characteristic of INH-induced hepatotoxicity. Copyright © 2014 John Wiley & Sons, Ltd. PMID:25331106

Su, Yijing; Zhang, Yun; Chen, Mi; Jiang, Zhenzhou; Sun, Lixin; Wang, Tao; Zhang, Luyong

2014-12-01

236

Bitter gourd suppresses lipopolysaccharide-induced inflammatory responses.  

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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

2008-06-11

237

Molecular Characterization of Lipopolysaccharide Binding to Human ?-1-Acid Glycoprotein.  

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The ability of AGP to bind circulating lipopolysaccharide (LPS) in plasma is believed to help reduce the proinflammatory effect of bacterial lipid A molecules. Here, for the first time we have characterized human AGP binding characteristics of the LPS from a number of pathogenic Gram-negative bacteria: Escherichia coli, Salmonella typhimurium, Klebsiella pneumonia, Pseudomonas aeruginosa, and Serratia marcescens. The binding affinity and structure activity relationships (SAR) of the AGP-LPS interactions were characterized by surface plasma resonance (SPR). In order to dissect the contribution of the lipid A, core oligosaccharide and O-antigen polysaccharide components of LPS, the AGP binding affinity of LPS from smooth strains, were compared to lipid A, Kdo2-lipid A, R(a), R(d), and R(e) rough LPS mutants. The SAR analysis enabled by the binding data suggested that, in addition to the important role played by the lipid A and core components of LPS, it is predominately the unique species- and strain-specific carbohydrate structure of the O-antigen polysaccharide that largely determines the binding affinity for AGP. Together, these data are consistent with the role of AGP in the binding and transport of LPS in plasma during acute-phase inflammatory responses to invading Gram-negative bacteria. PMID:23316371

Huang, Johnny X; Azad, Mohammad A K; Yuriev, Elizabeth; Baker, Mark A; Nation, Roger L; Li, Jian; Cooper, Matthew A; Velkov, Tony

2012-01-01

238

Zingerone attenuates lipopolysaccharide-induced acute lung injury in mice.  

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Zingerone, one of the active components of ginger, is a phenolic alkanone with antioxidant and anti-inflammatory properties. In the present study, we analyzed the role of zingerone against RAW 264.7 cells and acute lung injury induced by lipopolysaccharide (LPS) in mice. RAW cells or BALB/c mice were pretreated with zingerone one hour before stimulated with LPS. We found that zingerone significantly inhibited the production of LPS-induced proinflammatory cytokines in vitro and in vivo. When pretreated with zingerone, pulmonary histopathologic changes, as well as alveolar hemorrhage and neutrophil infiltration were substantially suppressed in lung tissues, with evidence of reduced myeloperoxidase (MPO) activity in murine acute lung injury model. The lung wet-to-dry weight (W/D) ratios, as the index of pulmonary edema, were markedly decreased by zingerone pretreatment. Furthermore, we demonstrated that zingerone attenuates the mitogen-activated protein kinases (MAPK) and nuclear factor-kappaB (NF-?B) signaling pathways through blocking the phosphorylation of ERK, p38/MAPK and I?B?, NF-?B/P65. These results suggest that zingerone may provide protective effects against LPS-induced ALI. PMID:24412620

Xie, Xianxing; Sun, Shicheng; Zhong, Weiting; Soromou, Lanan Wassy; Zhou, Xuan; Wei, Miaomiao; Ren, Yanling; Ding, Yu

2014-03-01

239

Antioxidant Effects of Cranberry Powder in Lipopolysaccharide Treated Hypercholesterolemic Rats  

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This study was conducted to investigate the effects of cranberry power on antioxidant defense system in rats fed an atherogenic diet and injected with lipopolysaccharide (LPS). Sprague-Dawley rats were divided into the following 5 groups: normal diet+saline (NS), atherogenic diet+saline (AS), atherogenic diet+LPS (AL), atherogenic diet with 5% cranberry powder+LPS (AL-C5), and atherogenic diet with 10% cranberry powder+LPS (AL-C10). Total antioxidant status measured by ferric reducing ability of plasma (FRAP) was significantly reduced by LPS injection (24%) and was restored by the cranberry powder treatment (P<0.05). In addition, the mean level of plasma total phenolics was significantly decreased by LPS injection (P<0.05) and tended to be increased when cranberry powder was incorporated in to the diet. Activity of serum superoxide dismutase (SOD) tended to be lowered by LPS injection and declined further in cranberry powder fortified groups. Overall results indicate that dietary cranberry powder may provide appropriate antioxidants to counter the diminished antioxidant status induced by exposing hypercholesterolemic rats to LPS. PMID:25054105

Kim, Mi Joung; Kim, Jung Hee; Kwak, Ho-Kyung

2014-01-01

240

Visualization and analysis of lipopolysaccharide distribution in binary phospholipid bilayers  

Energy Technology Data Exchange (ETDEWEB)

Lipopolysaccharide (LPS) is an endotoxin released from the outer membrane of Gram-negative bacteria during infections. It have been reported that LPS may play a role in the outer membrane of bacteria similar to that of cholesterol in eukaryotic plasma membranes. In this article we compare the effect of introducing LPS or cholesterol in liposomes made of dipalmitoylphosphatidylcholine/dioleoylphosphatidylcholine on the solubilization process by Triton X-100. The results show that liposomes containing LPS or cholesterol are more resistant to solubilization by Triton X-100 than the binary phospholipid mixtures at 4 {sup o}C. The LPS distribution was analyzed on GUVs of DPPC:DOPC using FITC-LPS. Solid and liquid-crystalline domains were visualized labeling the GUVs with LAURDAN and GP images were acquired using a two-photon microscope. The images show a selective distribution of LPS in gel domains. Our results support the hypothesis that LPS could aggregate and concentrate selectively in biological membranes providing a mechanism to bring together several components of the LPS-sensing machinery.

Henning, Maria Florencia [Instituto de Investigaciones Bioquimicas La Plata (INIBIOLP), CCT-La Plata, CONICET, Facultad de Ciencias Medicas, UNLP, Calles 60 y 120, 1900 La Plata (Argentina); Sanchez, Susana [Laboratory for Fluorescence Dynamics, University of California-Irvine, Irvine, CA (United States); Bakas, Laura, E-mail: lbakas@biol.unlp.edu.ar [Instituto de Investigaciones Bioquimicas La Plata (INIBIOLP), CCT-La Plata, CONICET, Facultad de Ciencias Medicas, UNLP, Calles 60 y 120, 1900 La Plata (Argentina); Departamento de Ciencias Biologicas, Facultad de Ciencias Exactas, UNLP, Calles 47 y 115, 1900 La Plata (Argentina)

2009-05-22

 
 
 
 
241

Trafficking of Shigella lipopolysaccharide in polarized intestinal epithelial cells.  

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Bacterial lipopolysaccharide (LPS) at the apical surface of polarized intestinal epithelial cells was previously shown to be transported from the apical to the basolateral pole of the epithelium (Beatty, W.L., and P.J. Sansonetti. 1997. Infect. Immun. 65:4395-4404). The present study was designed to elucidate the transcytotic pathway of LPS and to characterize the endocytic compartments involved in this process. Confocal and electron microscopic analyses revealed that LPS internalized at the apical surface became rapidly distributed within endosomal compartments accessible to basolaterally internalized transferrin. This compartment largely excluded fluid-phase markers added at either pole. Access to the basolateral side of the epithelium subsequent to trafficking to basolateral endosomes occurred via exocytosis into the paracellular space beneath the intercellular tight junctions. LPS appeared to exploit other endocytic routes with much of the internalized LPS recycled to the original apical membrane. In addition, analysis of LPS in association with markers of the endocytic network revealed that some LPS was sent to late endosomal and lysosomal compartments. PMID:10330399

Beatty, W L; Méresse, S; Gounon, P; Davoust, J; Mounier, J; Sansonetti, P J; Gorvel, J P

1999-05-17

242

Atmospheric pressure plasma treatment of lipopolysaccharide in a controlled environment  

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The atmospheric pressure plasma jet (APPJ) has been widely investigated for sterilization of surfaces, but studies on surface chemical changes of model compounds in controlled environments have been lacking. We present measurements on lipopolysaccharide (LPS) using x-ray photoelectron spectroscopy after 1% O2 in Ar APPJ treatments in controlled ambients composed of N2/Ar mixtures. By varying the N2 concentration from 20% to 100%, we find that the interaction of the jet with the environment plays a major role in modifying surface reactions. This is due to the plasma exciting N2, which quenches reactive oxygen species (ROS) that would otherwise modify the film surface. By minimizing the interaction of the APPJ with the environment, e.g. by changing the APPJ geometry, we show that surface modifications increase even when the plasma itself is removed farther from the LPS surface. Measurements on the biological activity, optical emission, and ozone production of the jet using O2, N2 and O2/N2 admixtures all demonstrate that ROS are readily quenched by N2 species excited by the plasma. These results clearly reveal the importance of considering plasma–environment interactions for APPJ treatments of surfaces. (fast track communication)

243

Complement activation by Proteus mirabilis negatively charged lipopolysaccharides.  

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Proteus mirabilis strains are human pathogens responsible for urinary tract infections and bacteremias and may be involved in rheumatoid arthritis. Lipopolysaccharide (LPS, bacterial endotoxin), the major component of the cell wall, is one of the virulence factors of Proteus. In the presented studies, we have investigated complement activation by LPSs isolated from P. mirabilis O10, O23, O30, and O43 strains, which differ in the number of negative COO- groups on their polysaccharide components. Four P. mirabilis strains studied were sensitive to complement-mediated killing, despite complement binding by their LPSs. The optimal complement binding by LPSs was detected in serum with functional assays for both the classical and alternative pathways. Complement activation in 80% serum by the smooth, uronic acid, and hexosamine containing P. mirabilis LPSs was not critically determined by the structure of their O-chain polysaccharides. One of four LPSs used as a model, P. mirabilis O10 LPS, fragmented C3 in an LPS dose- and time-dependent manner. It was detected by crossed-immunoelectrophoresis and capture ELISA with anti-C3c antibodies. The lower complement activation by 023 LPS correlates with its reduced C3 fragmentation, compared with three other Proteus LPSs studied. Rabbit anti-O antibodies enhanced the complement binding and factor C3 fragmentation by O10, O23, O30, and O43 P. mirabilis LPSs. PMID:11052177

Kaca, W; Literacka, E; Sjöholm, A G; Weintraub, A

2000-01-01

244

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

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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

2010-11-01

245

Pasteurella multocida Heddleston serovars 1 and 14 express different lipopolysaccharide structures but share the same lipopolysaccharide biosynthesis outer core locus.  

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Pasteurella multocida strains are classified using the Heddleston lipopolysaccharide (LPS) serotyping scheme into 16 serovars. Understanding the structural and genetic basis for this LPS typing scheme is important because protection against infections caused by P. multocida is generally considered to be serovar specific. Here we show that the serovar 14 type strain P2225 and the serovar 1 strains X73 and VP161 express similar LPS structures. However, the serovar 14 LPS lacks the terminal phosphocholine (PCho) residues present on the serovar 1 LPS and contains the 1,4-linked ?-galactose but not the 1,6-linked ?-galactose. Sequencing analysis of the LPS biosynthesis outer core loci of P2225 and the serovar 1 type strain X73 showed that they were nearly identical. However, the phosphocholine biosynthesis gene, pcgA of P2225 contained a 19bp nucleotide deletion. Complementation of P2225 with an intact pcgA resulted in an LPS structure identical to that expressed by serovar 1 strain VP161 and highly similar to that expressed by strain X73, with a 1,6-linked ?-galactose and both terminal PCho residues. This study has shown unequivocally that strains belonging to serovar 1 and 14 share a common LPS outer core locus and that minor changes within this locus can dramatically alter the LPS structure expressed on the surface of P. multocida, and thus has implications into our understanding of the potential to generate cross-protective vaccines. PMID:21371830

Harper, Marina; St Michael, Frank; John, Marietta; Vinogradov, Evgeny; Adler, Ben; Boyce, John D; Cox, Andrew D

2011-06-01

246

Btk Regulates Macrophage Polarization in Response to Lipopolysaccharide  

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Bacterial Lipopolysaccharide (LPS) is a strong inducer of inflammation and does so by inducing polarization of macrophages to the classic inflammatory M1 population. Given the role of Btk as a critical signal transducer downstream of TLR4, we investigated its role in M1/M2 induction. In Btk deficient (Btk?\\?) mice we observed markedly reduced recruitment of M1 macrophages following intraperitoneal administration of LPS. Ex vivo analysis demonstrated an impaired ability of Btk?/? macrophages to polarize into M1 macrophages, instead showing enhanced induction of immunosuppressive M2-associated markers in response to M1 polarizing stimuli, a finding accompanied by reduced phosphorylation of STAT1 and enhanced STAT6 phosphorylation. In addition to STAT activation, M1 and M2 polarizing signals modulate the expression of inflammatory genes via differential activation of transcription factors and regulatory proteins, including NF-?B and SHIP1. In keeping with a critical role for Btk in macrophage polarization, we observed reduced levels of NF-?B p65 and Akt phosphorylation, as well as reduced induction of the M1 associated marker iNOS in Btk?/? macrophages in response to M1 polarizing stimuli. Additionally enhanced expression of SHIP1, a key negative regulator of macrophage polarisation, was observed in Btk?/? macrophages in response to M2 polarizing stimuli. Employing classic models of allergic M2 inflammation, treatment of Btk?/? mice with either Schistosoma mansoni eggs or chitin resulted in increased recruitment of M2 macrophages and induction of M2-associated genes. This demonstrates an enhanced M2 skew in the absence of Btk, thus promoting the development of allergic inflammation. PMID:24465735

Ni Gabhann, Joan; Hams, Emily; Smith, Siobhan; Wynne, Claire; Byrne, Jennifer C.; Brennan, Kiva; Spence, Shaun; Kissenpfennig, Adrien; Johnston, James A.; Fallon, Padraic G.; Jefferies, Caroline A.

2014-01-01

247

Prolonged Neuroinflammation after Lipopolysaccharide Exposure in Aged Rats  

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Inflammation is a hallmark of several disease states ranging from neurodegeneration to sepsis but is also implicated in physiological processes like ageing. Non-resolving inflammation and prolonged neuroinflammation are unclear processes implicated in several conditions, including ageing. In this study we studied the long-term effects of endotoxemia, as systemic lipopolysaccharide (LPS) injection, focusing on the role of astrocyte activation and cytokine release in the brain of aged rats. A single dose of LPS (2 mg/kg) or 0.9% saline was injected intraperitoneally in aged rats. Levels of pro-inflammatory cytokines (TNF? and IL-1?) and NF-?B p65 activation were measured systemically and in hippocampal tissue. Astrocytes and cytokines release in the CNS were detected via double immunofluorescence staining at different time-points up to day 30. Serum levels of TNF? and IL-1? were significantly increased acutely after 30 minutes (p<0.001) and up to 6 hours (p<0.001) following LPS-injection. Centrally, LPS-treated rats showed up-regulated mRNA expression and protein levels of pro-inflammatory cytokines in the hippocampus. These changes associated with astrogliosis in the hippocampus dentate gyrus (DG), IL-1? immunoreactivity and elevated NF-?B p65 expression up to day 30 post LPS exposure. Overall, these data demonstrate that LPS induces prolonged neuroinflammation and astrocyte activation in the hippocampus of aged rats. Hippocampal NF-?B p65 and excessive astrocytes-derived IL-1? release may play a pivotal role in regulating long-lasting neuroinflammation. PMID:25170959

Fu, Hui Qun; Yang, Ting; Xiao, Wei; Fan, Long; Wu, Yan; Terrando, Niccolo; Wang, Tian Long

2014-01-01

248

Intermittent fasting attenuates lipopolysaccharide-induced neuroinflammation and memory impairment  

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Background Systemic bacterial infections often result in enduring cognitive impairment and are a risk factor for dementia. There are currently no effective treatments for infection-induced cognitive impairment. Previous studies have shown that intermittent fasting (IF) can increase the resistance of neurons to injury and disease by stimulating adaptive cellular stress responses. However, the impact of IF on the cognitive sequelae of systemic and brain inflammation is unknown. Methods Rats on IF for 30 days received 1 mg/kg of lipopolysaccharide (LPS) or saline intravenously. Half of the rats were subjected to behavioral tests and the other half were euthanized two hours after LPS administration and the hippocampus was dissected and frozen for analyses. Results Here, we report that IF ameliorates cognitive deficits in a rat model of sepsis by a mechanism involving NF-?B activation, suppression of the expression of pro-inflammatory cytokines, and enhancement of neurotrophic support. Treatment of rats with LPS resulted in deficits in cognitive performance in the Barnes maze and inhibitory avoidance tests, without changing locomotor activity, that were ameliorated in rats that had been maintained on the IF diet. IF also resulted in reduced levels of mRNAs encoding the LPS receptor TLR4 and inducible nitric oxide synthase (iNOS) in the hippocampus. Moreover, IF prevented LPS-induced elevation of IL-1?, IL-1? and TNF-? levels, and prevented the LPS-induced reduction of BDNF levels in the hippocampus. IF also significantly attenuated LPS-induced elevations of serum IL-1?, IFN-?, RANTES, TNF-? and IL-6 levels. Conclusions Taken together, our results suggest that IF induces adaptive responses in the brain and periphery that can suppress inflammation and preserve cognitive function in an animal model of systemic bacterial infection. PMID:24886300

2014-01-01

249

Sequence context induced antimicrobial activity: insight into lipopolysaccharide permeabilization.  

Science.gov (United States)

Lactoferrampin (WR17, Trp 268-Arg 284), an antimicrobial peptide, is known to have significant antibacterial and candidacidal activities. However, there are no previous studies explaining how WR17 permeabilizes the outer membrane of Gram negative bacteria and neutralizes endotoxins. In this study we used a series of assays like antimicrobial activity, calcein leakage, NPN dye uptake and endotoxin neutralization assay to show that the sequence context of WR17 modulates its multi-faceted activities. We determined the high resolution NMR structure of WR17 in LPS and found that the N-ter region forms a helix (Trp1-Phe11) and orients itself at an angle of 45° into the lipopolysaccharide (LPS) micelle, whereas the C-ter region (Lys13-Arg17) remains as a flexible extended random coil. We also verified this result through in silico molecular modeling simulation. Isothermal titration calorimetry showed that the interaction of WR17 and its analogues with LPS was primarily endothermic in nature. Using several fluorescence techniques such as anisotropy and red edge excitation shift assay we revealed motional restriction for Trp1 of WR17 in LPS. The distance between the indole ring of Trp1 of WR17 and the polar head group of LPS is around 7 Å, as obtained from the depth of insertion assay. Additionally, MD simulation demonstrated that the incorporation of the peptide in LPS is achieved with the help of the K(13)xK(15)xR(17) motif at the C-terminus. This novel anchoring "K(13)NKSR(17)" motif is currently being utilized in our ongoing research to design novel anti-endotoxic molecules. PMID:24714742

Ghosh, Anirban; Datta, Aritreyee; Jana, Jagannath; Kar, Rajiv Kumar; Chatterjee, Chiradip; Chatterjee, Subhrangsu; Bhunia, Anirban

2014-06-01

250

A new structural type for Haemophilus influenzae lipopolysaccharide. Structural analysis of the lipopolysaccharide from nontypeable Haemophilus influenzae strain 486.  

Science.gov (United States)

Structural elucidation of the sialylated lipopolysaccharide (LPS) of non-typeable Haemophilus influenzae (NTHi) strain 486 has been achieved by the application of high-field NMR techniques and ESI-MS along with composition and linkage analyses on O-deacylated LPS and oligosaccharide samples. It was found that the LPS contains the common element of H. influenzae, L-alpha-D-Hepp-(1-->2)-[PEtn-->6]-L-alpha-D-Hepp-(1-->3)-[beta-D-Glcp-(1-->4)]-L-alpha-D-Hepp-(1-->5)-[PPEtn-->4]-alpha-Kdop-(2-->6)-Lipid A, but instead of glycosyl substitution of the terminal heptose residue (HepIII) at the O2 position observed in other H. influenzae strains, HepIII is chain elongated at the O3 position by either lactose or sialyllactose (i.e. alpha-Neu5Ac-(2-->3)-beta-D-Galp-(1-->4)-beta-D-Glcp). The LPS is substituted by an O-acetyl group linked to the O2 position of HepIII and phosphocholine (PCho) which was located at the O6 position of a terminal alpha-D-Glcp residue attached to the central heptose, a molecular environment different from what has been reported earlier for PCho. In addition, minor substitution by O-linked glycine to the LPS was observed. By investigation of LPS from a lpsA mutant of NTHi strain 486, it was demonstrated that the lpsA gene product also is responsible for chain extension from HepIII in this strain. The involvement of lic1 in expression of PCho was established by investigation of a lic1 mutant of NTHi strain 486. PMID:11277939

Månsson, M; Bauer, S H; Hood, D W; Richards, J C; Moxon, E R; Schweda, E K

2001-04-01

251

Variation of Lipopolysaccharide among the Three Major Agrobacterium Species and the Effect of Environmental Stress on the Lipopolysaccharide Profile  

Directory of Open Access Journals (Sweden)

Full Text Available Lipopplysaccharide (LPS is a variable component among the bacterial species as wall as strains of a single species and this characteristic is helpful for discrimination between strains. However, we have only limited information about LPS variation and influence by environment in Agrobacterium strains. In this study, we analyzed variation of lipopolysaccharide (LPS among 34 Agrobacterium strains; 9 strains of A. tumefaciens, 15 strains of A. rhizogenes, 9 strains of A. vitis and one A. rubi strain. Most of the A. tumefaciens strains and every A. rhizogenes strains had high and low molecular weight LPS molecules (LPS I and LPS II, respectively. On the contrary, every A. vitis strains and two exceptional A. tumefaciens strains lacked LPS I but had a single LPS II band. The LPS profiles were stable phenotype in the Agrobacterium strains. Abiotic stresses such as high salinity, high and low pH and high and low temperature were given to representative strains in each species. Only a little alternation in the LPS profiles was observed under the stress conditions except the high temperature to LPS I. Cultivation at 35°C or higher resulted in a significant size reduction of LPS I in A. tumefaciens C58 strain down to the size similar to that of LPS II which attenuated the tumor formation. On the contrary, cultivation at the high temperature induced the exceptional A. tumefaciens strain MAFF 03-01001 to synthesize LPS I, which was absent at lower temperature in the strain. This phenomenon has never been observed so far at least in the family Rhizobiaceae.

H. H. Arafat

2009-01-01

252

Detectable serum flagellin and lipopolysaccharide and upregulated anti-flagellin and lipopolysaccharide immunoglobulins in human short bowel syndrome.  

Science.gov (United States)

Gut barrier dysfunction may occur in short bowel syndrome (SBS). We hypothesized that systemic exposure to flagellin and lipopolysaccharide (LPS) in SBS might regulate specific immune responses. We analyzed serial serum samples obtained from parenteral nutrition (PN)-dependent patients with SBS versus non-SBS control serum. Serum from 23 adult SBS patients was obtained at baseline and 4, 8, 12, 16, 20, and 24 wk in a trial of modified diet with or without growth hormone. Control serum was obtained from 48 healthy adults and 37 adults requiring PN during critical illness. Serum flagellin was detected by an ELISA recognizing an array of gram-negative flagellins, and LPS was detected by limulus assay. Serum flagellin- and LPS-specific immunoglobulin levels (IgM, IgA, and IgG) were determined by ELISA. Serum flagellin and LPS were undetectable in control subjects. In contrast, serum flagellin, LPS, or both were detected in 14 SBS patients (61%) during one or more time points [flagellin alone, 5/23 (22%); LPS alone, 6/23 (26%); or flagellin + LPS, 3/23 (13%)]. Flagellin-specific serum IgM, IgA, and IgG levels were markedly increased in SBS patients compared with both control populations and remained elevated during the 6-mo study period. LPS-specific IgA was significantly higher in SBS patients compared with healthy controls; LPS-specific IgM, IgA, and IgG levels each decreased over time in association with PN weaning. We conclude that adults with PN-dependent SBS are systemically exposed to flagellin and LPS, presumably from the gut lumen. This likely regulates innate and adaptive immune responses to these specific bacterial products. PMID:18003793

Ziegler, Thomas R; Luo, Menghua; Estívariz, Concepción F; Moore, Daniel A; Sitaraman, Shanthi V; Hao, Li; Bazargan, Niloofar; Klapproth, Jan-Michael; Tian, Junqiang; Galloway, John R; Leader, Lorraine M; Jones, Dean P; Gewirtz, Andrew T

2008-02-01

253

DMPD: Function of lipopolysaccharide (LPS)-binding protein (LBP) and CD14, thereceptor for LPS/LBP complexes: a short review. [Dynamic Macrophage Pathway CSML Database  

Full Text Available 1373512 Function of lipopolysaccharide (LPS)-binding protein (LBP) and CD14, thereceptor for LPS ... PS/LBP complexes: a short review. PubmedID 1373512 Title ... Function of lipopolysaccharide (LPS)-binding prote ...

254

Lipopolysaccharide profiles from nodules as markers of bradyrhizobium strains nodulating wild legumes.  

Science.gov (United States)

To develop the use of electrophoretic lipopolysaccharide profiles for Bradyrhizobium strain identification, we studied the feasibility of using electrophoresis of whole legume nodule homogenates to obtain distinctive lipopolysaccharide profiles. The electrophoretic patterns were the same whether we used nodule extracts, bacteroids, or cultured bacteria as samples, and there was no evidence of changes in the ladder-like pattern during the nodulation process. To assess the reliability of using lipopolysaccharide profiling performed with individual nodules for studying the diversity and microdistribution of the rhizobia nodulating wild shrub legumes, we used a population of Adenocarpus foliolosus seedlings. We obtained 75 different profiles from the 147 nodules studied. There was no dominant profile in the sample, and a plant with different nodules generally produced different profiles. Electrophoresis of legume root nodules proved to be a fast and discriminating technique for determining the diversity of a bradyrhizobial population, although it did not allow the genetic relationships among the nodulating strains to be studied. PMID:16349529

Santamaría, M; Gutiérrez-Navarro, A M; Corzo, J

1998-03-01

255

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

256

Periodontal ligament and gingival fibroblasts participate in the production of TGF-?, interleukin (IL)-8 and IL-10  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english The aim of this study was to quantify and compare the production of transforming growth factor beta (TGF-?), interleukin (IL)-8 and IL-10 by human cultured periodontal ligament and gingival fibroblasts both obtained from the same donors challenged with lipopolysaccharide (LPS) from Porphyromonas gin [...] givalis. Fibroblasts were exposed to 0.1-10 µg/mL of LPS from P. gingivalis and after 24 h the supernatants were collected and analyzed by enzyme-linked immunosorbent assay (ELISA). TGF-? protein production was upregulated in a concentration-dependent manner, mainly in gingival fibroblasts, which was statistically significant when challenged by 10 µg/mL LPS. Additionally, at this concentration, gingival fibroblasts had almost a two-fold increase in the amount of TGF-? when compared to periodontal ligament fibroblasts. Both periodontal ligament and gingival fibroblasts showed an increase in IL-8 production when challenged with 1 µg/mL and 10 µg/mL LPS. IL-10 production remained unaffected when challenged by any of the LPS concentrations tested in either periodontal ligament or gingival fibroblasts. Our results demonstrate that periodontal ligament and gingival fibroblasts when challenged by LPS from P. gingivalis with 24 h may play a critical role in producing TGF-? and IL-8 but not IL-10.

Ana Carolina de Faria, Morandini; Carla Renata, Sipert; Erivan Schnaider, Ramos-Junior; Daniel Thomas, Brozoski; Carlos Ferreira, Santos.

257

Protection against experimental melioidosis following immunisation with a lipopolysaccharide-protein conjugate.  

Science.gov (United States)

Melioidosis is a severe infectious disease caused by Burkholderia pseudomallei. It is refractory to antibiotic treatment and there is currently no licensed vaccine. In this report we detail the construction and protective efficacy of a polysaccharide-protein conjugate composed of B. pseudomallei lipopolysaccharide and the Hc fragment of tetanus toxin. Immunisation of mice with the lipopolysaccharide-conjugate led to significantly reduced bacterial burdens in the spleen 48 hours after challenge and afforded significant protection against a lethal challenge with B. pseudomallei. The conjugate generated significantly higher levels of antigen-specific IgG1 and IgG2a than in lipopolysaccharide-immunised mice. Immunisation with the conjugate also demonstrated a bias towards Th1 type responses, evidenced by high levels of IgG2a. In contrast, immunisation with unconjugated lipopolysaccharide evoked almost no IgG2a demonstrating a bias towards Th2 type responses. This study demonstrates the effectiveness of this approach in the development of an efficacious and protective vaccine against melioidosis. PMID:24892035

Scott, Andrew E; Ngugi, Sarah A; Laws, Thomas R; Corser, David; Lonsdale, Claire L; D'Elia, Riccardo V; Titball, Richard W; Williamson, E Diane; Atkins, Timothy P; Prior, Joann L

2014-01-01

258

DMPD: Lipopolysaccharide-binding molecules: transporters, blockers and sensors. [Dynamic Macrophage Pathway CSML Database  

Full Text Available 15241548 Lipopolysaccharide-binding molecules: transporters, blockers and sensors. Chaby R. Cell ... Mol Life Sci . 2004 Jul;61(14):1697-713. (.png) (.svg) (.html) ( ... ensors. Authors Chaby R. Publication Cell Mol Life Sci . 2004 Jul;61(14):1697-713. Pathway - PNG File (.pn ...

259

Protection against Experimental Melioidosis following Immunisation with a Lipopolysaccharide-Protein Conjugate  

Science.gov (United States)

Melioidosis is a severe infectious disease caused by Burkholderia pseudomallei. It is refractory to antibiotic treatment and there is currently no licensed vaccine. In this report we detail the construction and protective efficacy of a polysaccharide-protein conjugate composed of B. pseudomallei lipopolysaccharide and the Hc fragment of tetanus toxin. Immunisation of mice with the lipopolysaccharide-conjugate led to significantly reduced bacterial burdens in the spleen 48 hours after challenge and afforded significant protection against a lethal challenge with B. pseudomallei. The conjugate generated significantly higher levels of antigen-specific IgG1 and IgG2a than in lipopolysaccharide-immunised mice. Immunisation with the conjugate also demonstrated a bias towards Th1 type responses, evidenced by high levels of IgG2a. In contrast, immunisation with unconjugated lipopolysaccharide evoked almost no IgG2a demonstrating a bias towards Th2 type responses. This study demonstrates the effectiveness of this approach in the development of an efficacious and protective vaccine against melioidosis. PMID:24892035

Scott, Andrew E.; Ngugi, Sarah A.; Laws, Thomas R.; Corser, David; Lonsdale, Claire L.; D'Elia, Riccardo V.; Titball, Richard W.; Williamson, E. Diane; Atkins, Timothy P.; Prior, Joann L.

2014-01-01

260

NMDA receptor activation mediates lipopolysaccharide-induced depressive-like behavior  

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Full Text Available Background : Inflammation associated with cancer and induced by cancer therapy is associated with clinical signs of sickness behavior that can culminate in the development of symptoms of depression. Intraperitoneal administration of lipopolysaccharide to mice induces depressive-like behavior. Lipopolysaccharide-induced depressive-like behavior is mediated by activation of indoleamine 2,3-dioxygenase (IDO that degrades tryptophan along the kynurenine pathway and produces kynurenine metabolites such as quinolinic acid that acts as a N-Methyl-D-aspartate (NMDA receptor agonist. The present study was carried out to determine whether the NMDA receptor antagonist ketamine alleviates lipopolysaccharide-induced depressive-like behavior. Methods : Mice were injected intraperitoneally with ketamine or saline immediately before administration of the cytokine inducer lipopolysaccharide or saline via the same route. Their behavior and bodyweight were monitored up to 28 h post-injection. Depressive-like behavior was measured by increased immobility in the forced swim test and decreased sucrose preference for a sucrose solution. Brain, liver and plasma were collected 6 h and 28 h after treatment to measure biomarkers of inflammation. Results : Lipopolysaccharide induced the expression of cytokine, IDO, tryptophan 2,3-dioxygenase (TDO and heme oxygenase-1 at the periphery and in the brain. This effect was not altered by ketamine pretreatment. Ketamine blocked the development of depressive-like behavior but had no effect on sickness behavior measured by body weight loss, reduced food intake and decreased motor activity. Conclusions : These data indicate that ketamine is able to abrogate inflammation-induced depressive-like behavior by antagonising the activating effects of kynurenine metabolites on NMDA receptors.

Adam Walker

2012-09-01

 
 
 
 
261

Cerium dioxide nanoparticles do not modulate the lipopolysaccharide-induced inflammatory response in human monocytes  

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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 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 CeO2 nanoparticles. We investigated the inflammation-modulating effects of CeO2 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 µg/mL of CeO2 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: CeO2 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 CeO2 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 CeO2 nanoparticle-treated cells and control cells at noncytotoxic doses. No significant effects of CeO2 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 CeO2 nanoparticles followed by lipopolysaccharide exposure.Conclusion: CeO2 nanoparticles at noncytotoxic concentrations neither modulate pre-existing inflammation nor prime for subsequent exposure to lipopolysaccharides in human monocytes from healthy subjects.Keywords: cerium dioxide, nanoparticle, nanomedicine, inflammation, human monocyte, lipopolysaccharides

Hussain S

2012-03-01

262

Down-Modulation of L-Selectin by Lipopolysaccharide Is Not Required for Lipopolysaccharide-Induced Expression of CD14 in Mouse Bone Marrow Granulocytes  

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We established in previous studies that a constitutive lipopolysaccharide (LPS) receptor of low affinity is present on mouse bone marrow granulocytes (BMG). This yet-unidentified receptor is involved in the LPS-induced expression of a second LPS receptor, CD14. Because it has been claimed that L-selectin (CD62L) is a low-affinity LPS receptor in mature granulocytes (polymorphonuclear leukocytes), it may be asked whether this molecule could be the constitutive LPS receptor in BMG. We show in t...

Pe?dron, Thierry; Girard, Robert; Chaby, Richard

2001-01-01

263

Dose dependency and individual variability in selected clinical, haematological and blood biochemical responses after systemic lipopolysaccharide challenge in cattle  

DEFF Research Database (Denmark)

Previous studies have notede that susceptibility to systemic lipopolysaccharide (LPS) exposure seems to differ between individual cows. However, to date inter-individual variation in the existence or extent has never been backed up by statistical analyses.

Jacobsen, Stine; TØlbØll, Trine

2005-01-01

264

DMPD: Innate recognition of lipopolysaccharide by Toll-like receptor 4-MD-2. [Dynamic Macrophage Pathway CSML Database  

Full Text Available 15051069 Innate recognition of lipopolysaccharide by Toll-like receptor 4-MD-2 . Miyake K. Trends ... Microbiol. 2 004 Apr;12 (4):186-92 . (.png) (.svg) (.html) (.csml) ... n of lipopolysaccharide by Toll-like receptor 4-MD-2 . PubmedID 15051069 Title Innate recognition of lip ... opolysaccharide by Toll-like receptor 4-MD-2 . Authors Miyake K. Publication Trends Microbiol. 2 ... 004 Apr;12 (4):186-92 . Pathway - PNG File (.png) SVG File (.sv ...

265

Erythromycin inhibition of lipopolysaccharide-stimulated tumor necrosis factor alpha production by human monocytes in vitro.  

Science.gov (United States)

The mechanism of clinical effectiveness of low-dose and long-term erythromycin (EM) treatment for diffuse panbronchiolitis, sinobronchial syndrome, and associated otitis media with effusion was investigated by studying the effects of EM on tumor necrosis factor alpha (TNF-alpha) production by cultured human monocytes stimulated with lipopolysaccharide. At concentrations of 0.1 microgram/mL or more, EM inhibited TNF-alpha release from human monocytes stimulated by lipopolysaccharide in a dose-dependent manner. Of the other macrolides tested, roxithromycin, an EM derivative, also showed significant inhibition of TNF-alpha production, whereas josamycin failed to inhibit TNF-alpha release from monocytes. Nonmacrolidic drugs such as minocycline hydrochloride, ofloxacin, or penicillin G had no significant effect on TNF-alpha production. These results suggest that the clinical improvement of chronic respiratory diseases by EM may depend on the suppression of production of inflammatory cytokines such as TNF-alpha. PMID:1416647

Iino, Y; Toriyama, M; Kudo, K; Natori, Y; Yuo, A

1992-10-01

266

Olive oil phenolic compounds decrease the postprandial inflammatory response by reducing postprandial plasma lipopolysaccharide levels.  

Science.gov (United States)

We investigated the molecular mechanisms by which phenolic compounds (phenols) in virgin olive oil reduce the postprandial inflammatory response with the aim of identifying the transcription factor involved and the downstream effects. Olive oil-based breakfasts prepared with virgin olive oil (VOO) with high (398 ppm), intermediate (149 ppm) and low (70 ppm) phenol content were administered to 49 metabolic syndrome patients following a randomized crossover design. The consumption of a high-phenol VOO-based breakfast limited the increase of lipopolysaccharide plasma levels, TLR4, and SOCS3 proteins (poil, respectively), IL1B (p=0.002, intermediate oil), and CXCL1 (p=0.001) postprandial gene expression, in peripheral blood mononuclear cells, as compared with the consumption of a breakfast prepared with the same oil but with low or intermediate phenol content. Virgin olive oil phenolic compounds reduce the postprandial inflammatory response in association with postprandial plasma lipopolysaccharide levels. PMID:24874372

Camargo, Antonio; Rangel-Zuñiga, Oriol Alberto; Haro, Carmen; Meza-Miranda, Eliana Romina; Peña-Orihuela, Patricia; Meneses, Maria Eugenia; Marin, Carmen; Yubero-Serrano, Elena Maria; Perez-Martinez, Pablo; Delgado-Lista, Javier; Fernandez-Real, Jose Manuel; Luque de Castro, M Dolores; Tinahones, Francisco Jose; Lopez-Miranda, Jose; Perez-Jimenez, Francisco

2014-11-01

267

Transcriptional Activation of Mucin by Pseudomonas aeruginosa Lipopolysaccharide in the Pathogenesis of Cystic Fibrosis Lung Disease  

Science.gov (United States)

An unresolved question in cystic fibrosis (CF) research is how mutations of the CF transmembrane conductance regulator, a CI ion channel, cause airway mucus obstruction leading to fatal lung disease. Recent evidence has linked the CF transmembrane conductance regulator mutation to the onset and persistence of Pseudomonas aeruginosa infection in the airways, and here we provide evidence directly linking P. aeruginosa infection to mucus overproduction. We show that P. aeruginosa lipopolysaccharide profoundly upregulates transcription of the mucin gene MUC 2 in epithelial cells via inducible enhancer elements and that this effect is blocked by the tyrosine kinase inhibitors genistein and tyrphostin AG 126. These findings improve our understanding of CF pathogenesis and suggest that the attenuation of mucin production by lipopolysaccharide antagonists and tyrosine kinase inhibitors could reduce morbidity and mortality in this disease.

Li, Jian-Dong; Dohrman, Austin F.; Gallup, Marianne; Miyata, Susumu; Gum, James R.; Kim, Young S.; Nadel, Jay A.; Prince, Alice; Basbaum, Carol B.

1997-02-01

268

Effect of Kramecyne on the Inflammatory Response in Lipopolysaccharide-Stimulated Peritoneal Macrophages  

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Kramecyne is a new peroxide, it was isolated from Krameria cytisoides, methanol extract, and this plant was mostly found in North and South America. This compound showed potent anti-inflammatory activity; however, the mechanisms by which this compound exerts its anti-inflammatory effect are not well understood. In this study, we examined the effects of kramecyne on inflammatory responses in mouse lipopolysaccharide- (LPS-) induced peritoneal macrophages. Our findings indicate that kramecyne i...

Amp Xe Nchez-miranda, E. S.; Lemus-bautista, J.; Amp Xe Rez, S. P.; Amp Xe Rez-ramos, J. P.

2013-01-01

269

Renal Hemodynamic, Inflammatory, and Apoptotic Responses to Lipopolysaccharide in HO-1?/? Mice  

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Lipopolysaccharide (LPS) induces the stress-responsive gene heme oxygenase-1 (HO-1). The present study examined the significance of HO-1 in response to LPS. In HO-1?/? mice, as compared with HO-1+/+ mice, LPS provoked a greater reduction in glomerular filtration rate and renal blood flow, increased renal cytokine expression, and increased activation of nuclear factor (NF)-?B. Conversely, HO-1-overexpressing renal epithelial cells, exposed to LPS, exhibited a blunted activation of NF-?B ...

Tracz, Michal J.; Juncos, Julio P.; Grande, Joseph P.; Croatt, Anthony J.; Ackerman, Allan W.; Rajagopalan, Govindarajan; Knutson, Keith L.; Badley, Andrew D.; Griffin, Matthew D.; Alam, Jawed; Nath, Karl A.

2007-01-01

270

Artemisinin Attenuates Lipopolysaccharide-Stimulated Proinflammatory Responses by Inhibiting NF-?B Pathway in Microglia Cells  

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Microglial activation plays an important role in neuroinflammation, which contributes to neuronal damage, and inhibition of microglial activation may have therapeutic benefits that could alleviate the progression of neurodegeneration. Recent studies have indicated that the antimalarial agent artemisinin has the ability to inhibit NF-?B activation. In this study, the inhibitory effects of artemisinin on the production of proinflammatory mediators were investigated in lipopolysaccharide (LPS)-...

Zhu, Cansheng; Xiong, Zhaojun; Chen, Xiaohong; Peng, Fuhua; Hu, Xueqiang; Chen, Yanming; Wang, Qing

2012-01-01

271

Valproic Acid and Other HDAC Inhibitors Induce Microglial Apoptosis and Attenuate Lipopolysaccharide- induced Dopaminergic Neurotoxicity  

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Valproic acid (VPA), a widely prescribed drug for seizures and bipolar disorder, has been shown to be an inhibitor of histone deacetylase (HDAC). Our previous study has demonstrated that VPA pretreatment reduces lipopolysaccharide (LPS)-induced dopaminergic (DA) neurotoxicity through the inhibition of microglia over-activation. The aim of this study was to determine the mechanism underlying VPA-induced attenuation of microglia over-activation. Other HDAC inhibitors (HDACIs) were compared with...

Chen, Po See; Wang, Chao-chuan; Bortner, Carl D.; Peng, Giia-sheun; Wu, Xuefei; Pang, Hao; Lu, Ru-band; Gean, Po-wu; Chuang, De-maw; Hong, Jau-shyong

2007-01-01

272

LIPOPOLYSACCHARIDE ATTENUATES PHRENIC LONG-TERM FACILITATION FOLLOWING ACUTE INTERMITTENT HYPOXIA  

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Lipopolysaccharide (LPS) induces inflammatory responses, including microglial activation in the central nervous system. Since LPS impairs certain forms of hippocampal and spinal neuroplasticity, we hypothesized that LPS would impair phrenic long-term facilitation (pLTF) following acute intermittent hypoxia (AIH) in outbred Sprague-Dawley (SD) and inbred Lewis (L) rats.. Approximately three hours following a single LPS injection (i.p.), the phrenic response during hypoxic episodes is reduced i...

Vinit, Ste?phane; Windelborn, James A.; Mitchell, Gordon S.

2011-01-01

273

Hepatotoxic Interaction of Sulindac with Lipopolysaccharide: Role of the Hemostatic System  

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Sulindac (SLD) is a nonsteroidal anti-inflammatory drug (NSAID) that has been associated with a greater incidence of idiosyncratic hepatotoxicity in human patients than other NSAIDs. One hypothesis regarding idiosyncratic adverse drug reactions is that interaction of a drug with a modest inflammatory episode precipitates liver injury. In this study, we tested the hypothesis that lipopolysaccharide (LPS) interacts with SLD to cause liver injury in rats. SLD (50 mg/kg) or its vehicle was admini...

Zou, Wei; Devi, Sachin S.; Sparkenbaugh, Erica; Younis, Husam S.; Roth, Robert A.; Ganey, Patricia E.

2009-01-01

274

The effect of exogenous rhizobial lipopolysaccharide on symbiosis of Rhizobium leguminosarum bv. trifolii with red clover  

Directory of Open Access Journals (Sweden)

Full Text Available The effectivity of symbiosis of Rhizobium leguminosarum bv. trifolii with red clover in the presence of exogenous lipopolysaccharide (LPS preparation was measured as a yield of green mass of infected plants. The addition of complete LPS that had been obtained from homological Rhizobium strains influenced significantly the growth of plants. In the presence of defective LPS of Rhizobium mutant the effectivity of symbiosis did not change.

Maria G?owacka

1996-12-01

275

Effects of Superantigen and Lipopolysaccharide on Induction of CD80 through Apoptosis of Human Monocytes  

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To investigate the mechanisms underlying superantigen (SAg) stimulation, we analyzed the effect of SAg on monocyte responses with or without lipopolysaccharide (LPS). Addition of gamma interferon (IFN-?) to unstimulated cultures induced a marked increase in the number of CD80+ monocytes, which was inhibited by LPS through the action of interleukin-10. However, CD80+ monocytes began to increase before IFN-? production, observed after 9 h of stimulation with staphylococcal enterotoxin B (SEB)...

Takahashi, Masahiro; Takahashi, Maiko; Shinohara, Fumiaki; Takada, Haruhiko; Rikiishi, Hidemi

2001-01-01

276

Lipopolysaccharide structures of Campylobacter fetus are related to heat-stable serogroups.  

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To determine whether lipopolysaccharide (LPS) structures of Campylobacter fetus are related to the three known heat-stable serogroups, proteinase K-treated whole cell lysates obtained from strains of each serogroup were electrophoresed in polyacrylamide gels. All strains had smooth-type LPS with multiple high-molecular-weight repeating units. The profiles of serogroup A from C. fetus subsp. fetus and from C. fetus subsp. venerealis were identical, but they were different from those of C. fetu...

Perez-perez, G. I.; Blaser, M. J.; Bryner, J. H.

1986-01-01

277

?-PHENYL-N-TERT-BUTYL-NITRONE ATTENUATES LIPOPOLYSACCHARIDE- INDUCED NEURONAL INJURY IN THE NEONATAL RAT BRAIN  

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Although white matter damage is a fundamental neuropathological feature of periventricular leukomalacia (PVL), the motor and cognitive deficits observed later in infants with PVL indicate the possible involvement of cerebral neuronal dysfunction. Using a previously developed rat model of white matter injury induced by cerebral lipopolysaccharide (LPS) injection, we investigated whether LPS exposure also results in neuronal injury in the neonatal brain and whether ?-phenyl-n-tert-butyl-nitron...

Fan, Lir-wan; Mitchell, Helen J.; Rhodes, Philip G.; Cai, Zhengwei

2008-01-01

278

Unusual fatty acid substitution in lipids and lipopolysaccharides of Helicobacter pylori.  

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Cellular fatty acids, phospholipid fatty acids, and lipopolysaccharide fatty acids of four strains of Helicobacter pylori were analyzed by gas-liquid chromatography. The presence of myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, 19-carbon cyclopropane fatty acid, beta-hydroxypalmitic acid, and beta-hydroxystearic acid was confirmed. In phospholipids, myristic acid and 19-carbon cyclopropane fatty acid were the major fatty acids. Hydroxy fatty acids and unsaturated fatt...

Geis, G.; Leying, H.; Suerbaum, S.; Opferkuch, W.

1990-01-01

279

Hyphomonas spp., Shewanella spp., and Other Marine Bacteria Lack Heterogeneous (Ladderlike) Lipopolysaccharides  

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The lipopolysaccharides (LPS) of 19 marine bacteria were examined for size heterogeneities by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis in conjunction with an LPS-specific silver staining method. Fifteen marine bacteria had an R-type LPS instead of the ladderlike LPS array characteristic of most bacteria. In addition, three marine bacteria also had a single large LPS molecule. Without constraints (e.g., surface masking), R-type LPS, a more hydrophobic molecule, predomina...

Sledjeski, Darren D.; Weiner, Ronald M.

1991-01-01

280

Influence of the lipopolysaccharide structure of Salmonella enterica serovar Enteritidis on interactions with pig neutrophils  

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Abstract The key process for immune response development is the recognition of bacteria by the immune system of the host based on the sensing of pathogen-associated molecular patterns (PAMP). One of the most important PAMP is the lipopolysaccharide (LPS) molecule, a complex molecule present in the outer membrane of Gram negative bacteria. In this study we were interested in how different parts of the LPS of Salmonella enterica serovar Enteritidis are recognized by porcine neutrophi...

2011-01-01

 
 
 
 
281

Polymyxin B Inadequately Quenches the Effects of Contaminating Lipopolysaccharide on Murine Dendritic Cells  

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Dendritic cell (DC) activation is commonly used as a measure of the immunomodulatory potential of candidate exogenous and endogenous molecules. Residual lipopolysaccharide (LPS) contamination is a recurring theme and the potency of LPS is not always fully appreciated. To address this, polymyxin B (PmB) is often used to neutralise contaminating LPS. However, the limited capacity of this antibiotic to successfully block these effects is neglected. Therefore, this study aimed to determine the mi...

Tynan, Graham A.; Mcnaughton, Anne; Jarnicki, Andrew; Tsuji, Takao; Lavelle, Ed C.

2012-01-01

282

Complement activation by polysaccharide of lipopolysaccharide: an important virulence determinant of salmonellae.  

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Salmonellae with differences only in the O-antigenic polysaccharide of their lipopolysaccharide were previously shown to differentially activate complement via the alternative pathway, causing them to be ingested at different rates by the mouse macrophage-like cell line J774. We now show that this mechanism could explain the different virulence of these strains in vivo. Mouse peritoneal macrophages (thioglycolate induced) ingest these salmonellae at rates that are inversely proportional to th...

Liang-takasaki, C. J.; Saxe?n, H.; Ma?kela?, P. H.; Leive, L.

1983-01-01

283

Absence of a characteristic cell wall lipopolysaccharide in the phototrophic bacterium Chloroflexus aurantiacus.  

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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 characteristic LPS constituents, such as 3-hydroxy fatty acids, 2-keto-3-deoxyoctonate, heptoses, or O-chain sugars. Phenolchloroform-petroleum ether extracts were also free from precipitable LPS. A l...

Meissner, J.; Krauss, J. H.; Ju?rgens, U. J.; Weckesser, J.

1988-01-01

284

Lactoferrin Protects against Development of Hepatitis Caused by Sensitization of Kupffer Cells by Lipopolysaccharide  

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BALB/c mice were intravenously injected with lipopolysaccharide (LPS) (0.05 ?g/g of body weight) 7 days after being primed with zymosan. Recombinant human lactoferrin (250 ?g/g of body weight), intravenously administered 1 day before the injection of LPS, significantly lessened the severity of hepatitis, as assessed by levels of serum alanine transaminase compared to those seen when casein was administered. The transient rise of serum tumor necrosis factor alpha (TNF-?) after LPS treatment...

Yamaguchi, Makoto; Matsuura, Motoi; Kobayashi, Kiyoshi; Sasaki, Hajime; Yajima, Takaji; Kuwata, Tamotsu

2001-01-01

285

Engineering N-linked protein glycosylation with diverse O antigen lipopolysaccharide structures in Escherichia coli  

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Campylobacter jejuni has a general N-linked protein glycosylation system that can be functionally transferred to Escherichia coli. In this study, we engineered E. coli cells in a way that two different pathways, protein N-glycosylation and lipopolysaccharide (LPS) biosynthesis, converge at the step in which PglB, the key enzyme of the C. jejuni N-glycosylation system, transfers O polysaccharide from a lipid carrier (undecaprenyl pyrophosphate) to an acceptor protein. PglB was the only protein...

Feldman, Mario F.; Wacker, Michael; Hernandez, Marcela; Hitchen, Paul G.; Marolda, Cristina L.; Kowarik, Michael; Morris, Howard R.; Dell Anne; Valvano, Miguel A.; Aebi, Markus

2005-01-01

286

Enzymatic deacylation of the lipid A moiety of Salmonella typhimurium lipopolysaccharides by human neutrophils.  

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Lipid A, the toxic moiety of Gram-negative bacterial lipopolysaccharides (endotoxins), is a glucosamine disaccharide to which fatty acid and phosphate residues are covalently attached. Recent studies of Salmonella lipid A indicate that 3-hydroxytetradecanoic acid (3-OH-14:0) residues are directly linked to the glucosamine backbone and the nonhydroxylated fatty acids (principally dodecanoic and tetradecanoic acids) are esterified to the hydroxyl groups of some of the 3-OH-14:0 molecules. We re...

Hall, C. L.; Munford, R. S.

1983-01-01

287

Modulation of Lipopolysaccharide-Induced Monocyte Activation by Heparin-Binding Protein and Fucoidan  

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Activated polymorphonuclear leukocytes release heparin-binding protein (HBP; also known as CAP37 or azurocidin) from azurophilic granules. HBP is a strong chemoattractant for monocytes that also prolongs monocyte survival and potentiates endotoxin (lipopolysaccharide [LPS])-induced production of tumor necrosis factor alpha (TNF-?). We investigated the binding of fluorescein isothiocyanate-conjugated HBP to human monocytes in the presence of EDTA and the polysaccharide fucoidan. EDTA, which c...

Heinzelmann, Michael; Polk, Hiram C.; Miller, Frederick N.

1998-01-01

288

Presence or Absence of Lipopolysaccharide O Antigens Affects Type III Secretion by Pseudomonas aeruginosa?  

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Pseudomonas aeruginosa is one of the major causative agents of mortality and morbidity in hospitalized patients due to a multiplicity of virulence factors associated with both chronic and acute infections. Acute P. aeruginosa infection is primarily mediated by planktonic bacteria expressing the type III secretion system (TTSS), a surface-attached needle-like complex that injects cytotoxins directly into eukaryotic cells, causing cellular damage. Lipopolysaccharide (LPS) is the principal surfa...

Augustin, D. K.; Song, Y.; Baek, M. S.; Sawa, Y.; Singh, G.; Taylor, B.; Rubio-mills, A.; Flanagan, J. L.; Wiener-kronish, J. P.; Lynch, S. V.

2007-01-01

289

Effect of quercetin on lipopolysaccharide induced-sickness behavior and oxidative stress in rats  

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Objectives : Gram-negative infections and control infusion of recombinant cytokines in human have been shown to induce sickness behavior characterized by fever, prolong sleep, decreased food and water intake, reduced mobility, depression, and anxiety. Therefore, the present study was undertaken to investigate the effect of bioflavonoid quercetin in lipopolysaccharide (LPS)-induced sickness behavior. Materials and Methods : Wistar albino rats were divided into six groups (n=6). ...

Sah Sangeeta; Tirkey Naveen; Kuhad Anurag; Chopra Kanwaljit

2011-01-01

290

Isolation, purification, and partial characterization of a lipopolysaccharide from Haemophilus pleuropneumoniae.  

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Lipopolysaccharide (LPS) from Haemophilus pleuropneumoniae 1536, serotype 2, was isolated and purified by a procedure designed to be equally satisfactory for both smooth- and rough-type LPS. The LPS yield was 53%. Analysis of the preparations revealed that protein, nucleic acid, and cellular phospholipid contamination was negligible (less than 0.1%). Analysis of the sugar content of the LPS by gas-liquid chromatography and colorimetric analysis revealed the presence of rhamnose, mannose, gala...

Maudsley, J. R.; Kadis, S.; Mayberry, W. R.

1986-01-01

291

Lipopolysaccharide Enhances the Production of Vascular Endothelial Growth Factor by Human Pulp Cells in Culture  

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We investigated whether vascular endothelial growth factor (VEGF) production by human pulp cells (HPC) is regulated by lipopolysaccharide (LPS) in relation to the pathogenesis of pulpitis. Although HPC incubated with medium alone only marginally expressed VEGF mRNA and produced a low level of VEGF as detected by enzyme-linked immunosorbent assay, the VEGF mRNA expression and VEGF production were markedly enhanced upon stimulation with LPS from Escherichia coli. Prevotella intermedia LPS, phor...

Matsushita, K.; Motani, R.; Sakuta, T.; Nagaoka, S.; Matsuyama, T.; Abeyama, K.; Maruyama, I.; Takada, H.; Torii, M.

1999-01-01

292

Pasteurella multocida Heddleston Serovar 3 and 4 Strains Share a Common Lipopolysaccharide Biosynthesis Locus but Display both Inter- and Intrastrain Lipopolysaccharide Heterogeneity  

Science.gov (United States)

Pasteurella multocida is a Gram-negative multispecies pathogen and the causative agent of fowl cholera, a serious disease of poultry which can present in both acute and chronic forms. The major outer membrane component lipopolysaccharide (LPS) is both an important virulence factor and a major immunogen. Our previous studies determined the LPS structures expressed by different P. multocida strains and revealed that a number of strains belonging to different serovars contain the same LPS biosynthesis locus but express different LPS structures due to mutations within glycosyltransferase genes. In this study, we report the full LPS structure of the serovar 4 type strain, P1662, and reveal that it shares the same LPS outer core biosynthesis locus, L3, with the serovar 3 strains P1059 and Pm70. Using directed mutagenesis, the role of each glycosyltransferase gene in LPS outer core assembly was determined. LPS structural analysis of 23 Australian field isolates that contain the L3 locus revealed that at least six different LPS outer core structures can be produced as a result of mutations within the LPS glycosyltransferase genes. Moreover, some field isolates produce multiple but related LPS glycoforms simultaneously, and three LPS outer core structures are remarkably similar to the globo series of vertebrate glycosphingolipids. Our in-depth analysis showing the genetics and full range of P. multocida lipopolysaccharide structures will facilitate the improvement of typing systems and the prediction of the protective efficacy of vaccines. PMID:23974032

Harper, Marina; St. Michael, Frank; John, Marietta; Vinogradov, Evgeny; Steen, Jennifer A.; van Dorsten, Lieke; Steen, Jason A.; Turni, Conny; Blackall, Patrick J.; Adler, Ben; Cox, Andrew D.

2013-01-01

293

Pasteurella multocida Heddleston serovar 3 and 4 strains share a common lipopolysaccharide biosynthesis locus but display both inter- and intrastrain lipopolysaccharide heterogeneity.  

Science.gov (United States)

Pasteurella multocida is a Gram-negative multispecies pathogen and the causative agent of fowl cholera, a serious disease of poultry which can present in both acute and chronic forms. The major outer membrane component lipopolysaccharide (LPS) is both an important virulence factor and a major immunogen. Our previous studies determined the LPS structures expressed by different P. multocida strains and revealed that a number of strains belonging to different serovars contain the same LPS biosynthesis locus but express different LPS structures due to mutations within glycosyltransferase genes. In this study, we report the full LPS structure of the serovar 4 type strain, P1662, and reveal that it shares the same LPS outer core biosynthesis locus, L3, with the serovar 3 strains P1059 and Pm70. Using directed mutagenesis, the role of each glycosyltransferase gene in LPS outer core assembly was determined. LPS structural analysis of 23 Australian field isolates that contain the L3 locus revealed that at least six different LPS outer core structures can be produced as a result of mutations within the LPS glycosyltransferase genes. Moreover, some field isolates produce multiple but related LPS glycoforms simultaneously, and three LPS outer core structures are remarkably similar to the globo series of vertebrate glycosphingolipids. Our in-depth analysis showing the genetics and full range of P. multocida lipopolysaccharide structures will facilitate the improvement of typing systems and the prediction of the protective efficacy of vaccines. PMID:23974032

Harper, Marina; St Michael, Frank; John, Marietta; Vinogradov, Evgeny; Steen, Jennifer A; van Dorsten, Lieke; Steen, Jason A; Turni, Conny; Blackall, Patrick J; Adler, Ben; Cox, Andrew D; Boyce, John D

2013-11-01

294

Sirtuin inhibition attenuates the production of inflammatory cytokines in lipopolysaccharide-stimulated macrophages  

Energy Technology Data Exchange (ETDEWEB)

Highlights: Black-Right-Pointing-Pointer Lipopolysaccharide-stimulated macrophages were treated with cambinol and sirtinol. Black-Right-Pointing-Pointer Cambinol and sirtinol decreased lipopolysaccharide-induced cytokines. Black-Right-Pointing-Pointer Cambinol decreased NF-{kappa}B activity but had no impact on p38 MAPK activation. Black-Right-Pointing-Pointer Sirtuins are an interesting target for the treatment of inflammatory diseases. -- Abstract: In several inflammatory conditions such as rheumatoid arthritis or sepsis, the regulatory mechanisms of inflammation are inefficient and the excessive inflammatory response leads to damage to the host. Sirtuins are class III histone deacetylases that modulate the activity of several transcription factors that are implicated in immune responses. In this study, we evaluated the impact of sirtuin inhibition on the activation of lipopolysaccharide (LPS)-stimulated J774 macrophages by assessing the production of inflammatory cytokines. The pharmacologic inhibition of sirtuins decreased the production of tumour necrosis factor-alpha (TNF-{alpha}) interleukin 6 (IL-6) and Rantes. The reduction of cytokine production was associated with decreased nuclear factor kappa B (NF-{kappa}B) activity and inhibitor kappa B alpha (I{kappa}B{alpha}) phosphorylation while no impact was observed on the phosphorylation status of p38 mitogen-activated kinase (p38 MAPK). This work shows that sirtuin pharmacologic inhibitors are a promising tool for the treatment of inflammatory conditions.

Fernandes, Claudia A. [Universite catholique de Louvain, Louvain Drug Research Institute (LDRI), Pharmaceutics and Drug Delivery Research Group, Brussels B-1200 (Belgium); Fievez, Laurence [University of Liege, GIGA-Research, Laboratory of Cellular and Molecular Immunology, Liege B-4000 (Belgium); Neyrinck, Audrey M.; Delzenne, Nathalie M. [Universite catholique de Louvain, LDRI, Metabolism and Nutrition Research Group, Brussels B-1200 (Belgium); Bureau, Fabrice [University of Liege, GIGA-Research, Laboratory of Cellular and Molecular Immunology, Liege B-4000 (Belgium); Vanbever, Rita, E-mail: rita.vanbever@uclouvain.be [Universite catholique de Louvain, Louvain Drug Research Institute (LDRI), Pharmaceutics and Drug Delivery Research Group, Brussels B-1200 (Belgium)

2012-04-20

295

Sirtuin inhibition attenuates the production of inflammatory cytokines in lipopolysaccharide-stimulated macrophages  

International Nuclear Information System (INIS)

Highlights: ? Lipopolysaccharide-stimulated macrophages were treated with cambinol and sirtinol. ? Cambinol and sirtinol decreased lipopolysaccharide-induced cytokines. ? Cambinol decreased NF-?B activity but had no impact on p38 MAPK activation. ? Sirtuins are an interesting target for the treatment of inflammatory diseases. -- Abstract: In several inflammatory conditions such as rheumatoid arthritis or sepsis, the regulatory mechanisms of inflammation are inefficient and the excessive inflammatory response leads to damage to the host. Sirtuins are class III histone deacetylases that modulate the activity of several transcription factors that are implicated in immune responses. In this study, we evaluated the impact of sirtuin inhibition on the activation of lipopolysaccharide (LPS)-stimulated J774 macrophages by assessing the production of inflammatory cytokines. The pharmacologic inhibition of sirtuins decreased the production of tumour necrosis factor-alpha (TNF-?) interleukin 6 (IL-6) and Rantes. The reduction of cytokine production was associated with decreased nuclear factor kappa B (NF-?B) activity and inhibitor kappa B alpha (I?B?) phosphorylation while no impact was observed on the phosphorylation status of p38 mitogen-activated kinase (p38 MAPK). This work shows that sirtuin pharmacologic inhibitors are a promising tool for the treatment of inflammatory conditions.

296

Propolis antimicrobial activity against periodontopathic bacteria  

Directory of Open Access Journals (Sweden)

Full Text Available Propolis extract antimicrobial activity against periodontopathic (ATCC bacteria was investigated "in vitro". Bacterial strains tested were: Prevotella intermedia, Prevotella melaninogenica, Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Capnocytophaga gingivalis and Fusobacterium nucleatum. Minimal inhibitory concentration (MIC for the strains tested was determined using the method of broth dilution with the propolis extract in serial concentrations. Results showed MIC of 1 µg/ml for Actinobacillus actinomycetemcomitans and Capnocytophaga gingivalis; and 0.25 µg/ml for Prevotella intermedia, Prevotella melaninogenica, Porphyromonas gingivalis and Fusobacterium nucleatum. Some superinfectant organisms were also tested: Candida albicans susceptibility to propolis ethanolic extract was demonstrated at a concentration of 12 µg/ml. The MIC for Pseudomonas aeruginosa, Escherichia coli and Staphylococcus aureus (wild types was 14 µg/ml. All periodontal pathogens and superinfectants tested were susceptible to the propolis extract. The positive results suggest that the propolis extract should be further tested as an adjuvant to periodontal therapy.

Gebara Elaine C.E.

2002-01-01

297

A protein secretion system linked to bacteroidete gliding motility and pathogenesis  

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Porphyromonas gingivalis secretes strong proteases called gingipains that are implicated in periodontal pathogenesis. Protein secretion systems common to other Gram-negative bacteria are lacking in P. gingivalis, but several proteins, including PorT, have been linked to gingipain secretion. Comparative genome analysis and genetic experiments revealed 11 additional proteins involved in gingipain secretion. Six of these (PorK, PorL, PorM, PorN, PorW, and Sov) were similar in sequence to Flavoba...

Sato, Keiko; Naito, Mariko; Yukitake, Hideharu; Hirakawa, Hideki; Shoji, Mikio; Mcbride, Mark J.; Rhodes, Ryan G.; Nakayama, Koji

2010-01-01

298

Bacterial Induction of Beta Interferon in Mice Is a Function of the Lipopolysaccharide Component  

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We investigated the reason for the inability of lipopolysaccharide (LPS)-resistant (Lps-defective [Lpsd]) C57BL/10ScCr mice to produce beta interferon (IFN-?) when stimulated with bacteria. For this purpose, the IFN-? and other macrophage cytokine responses induced by LPS and several killed gram-negative and gram-positive bacteria in LPS-sensitive (Lps-normal [Lpsn]; C57BL/10ScSn and BALB/c) and Lpsd (C57BL/10ScCr and BALB/c/l) mice in vitro and in vivo were investigated on the mRNA and pro...

Sing, Andreas; Merlin, Thomas; Knopf, Hans-peter; Nielsen, Peter J.; Loppnow, Harald; Galanos, Chris; Freudenberg, Marina A.

2000-01-01

299

Protective Effect of Phosphatidylcholine on Lipopolysaccharide-Induced Acute Inflammation in Multiple Organ Injury  

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Soybean polyunsaturated phosphatidylcholine (PC) is thought to exert anti-inflammatory activities and has potent effects in attenuating acute renal failure and liver dysfunction. The aim of this study was to investigate the effects of PC in protecting multiple organ injury (MOI) from lipopolysaccharide (LPS). Six groups of rats (N=8) were used in this study. Three groups acted as controls and received only saline, hydrocortisone (HC, 6 mg/kg, i.v.) or PC (600 mg/kg, i.p.) without LPS (15 mg/k...

Jung, Yoon Yang; Nam, Yunsung; Park, Yong Seol; Lee, Ho Sung; Hong, Soon Auck; Kim, Beom Keun; Park, Eon Sub; Chung, Yoon Hee; Jeong, Ji Hoon

2013-01-01

300

Role of lipopolysaccharide and complement in susceptibility of Haemophilus ducreyi to human serum.  

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The role of lipopolysaccharide (LPS) in the susceptibility of Haemophilus ducreyi to human serum and the mechanism of complement activation by serum-susceptible (Sers) strains were investigated. Serum treated with 2 mM Mg2+ and 20 mM ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid was nonbactericidal, but inulin-treated serum remained bactericidal. Absorption of serum with heat-killed whole cells of an Sers strain removed its bactericidal activity against the absorbing s...

Wiseman, G. M.; Ronald, A. R.

1985-01-01

 
 
 
 
301

Control of lipopolysaccharide biosynthesis and release by Escherichia coli and Salmonella typhimurium.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The influence of the relA gene on lipopolysaccharide (LPS) biosynthesis and release by Escherichia coli and Salmonella typhimurium was investigated. Similar results were obtained with both species. The incorporation of [3H]galactose into LPS by galE mutants was inhibited by at least 50% (as compared with normal growing controls) during amino acid deprivation of relA+ strains. This inhibition could be prevented by the treatment of the amino acid-deprived relA+ bacteria with chloramphenicol, a ...

Ishiguro, E. E.; Vanderwel, D.; Kusser, W.

1986-01-01

302

The reaction of dental pulp to Escherichia coli lipopolysaccharide and Enterococcus faecalis lipoteichoic acid  

Directory of Open Access Journals (Sweden)

Full Text Available This research evaluates the effects of the lipopolysaccharides (LPS from Escherichia coli and lipoteichoic acid (LTA from Enterococcus faecalis on dental pulp. These molecules are components of the Gram-negative and Gram-positive bacteria cell wall, respectively. Ten dogs were used in the experiment. Inoculation in surgically opened pulp and coronal restoration with glass ionomer was the method chosen. The evaluation times were 1, 7, 15, 30 and 60 days. The results showed that the LPS and LTA, at 150 µg/ml, produced a negative interference in the pulp leading to destruction. LTA caused less damage than LPS.

Oliveira Laudimar Alves de

2003-01-01

303

Lipopolysaccharide Membranes and Membrane Proteins of Pseudomonas aeruginosa Studied by Computer Simulation  

Energy Technology Data Exchange (ETDEWEB)

Pseudomonas aeruginosa is a ubiquitous environmental Gram-negative bacterium with high metabolic versatility and an exceptional ability to adapt to a wide range of ecological environments, including soil, marches, coastal habitats, plant and animal tissues. Gram-negative microbes are characterized by the asymmetric lipopolysaccharide outer membrane, the study of which is important for a number of applications. The adhesion to mineral surfaces plays a central role in characterizing their contribution to the fate of contaminants in complex environmental systems by effecting microbial transport through soils, respiration redox chemistry, and ion mobility. Another important application stems from the fact that it is also a major opportunistic human pathogen that can result in life-threatening infections in many immunocompromised patients, such as lung infections in children with cystic fibrosis, bacteraemia in burn victims, urinary-tract infections in catheterized patients, hospital-acquired pneumonia in patients on respirators, infections in cancer patients receiving chemotherapy, and keratitis and corneal ulcers in users of extended-wear soft contact lenses. The inherent resistance against antibiotics which has been linked with the specific interactions in the outer membrane of P. aeruginosa makes these infections difficult to treat. Developments in simulation methodologies as well as computer hardware have enabled the molecular simulation of biological systems of increasing size and with increasing accuracy, providing detail that is difficult or impossible to obtain experimentally. Computer simulation studies contribute to our understanding of the behavior of proteins, protein-protein and protein-DNA complexes. In recent years, a number of research groups have made significant progress in applying these methods to the study of biological membranes. However, these applications have been focused exclusively on lipid bilayer membranes and on membrane proteins in lipid bilayers. A few simulation studies of outer membrane proteins of Gram-negative bacteria have been reported using simple lipid bilayers, even though this is not a realistic representation of the outer membrane environment. This contribution describes our recent molecular simulation studies of the rough lipopolysaccharide membrane of P. aeruginosa, which are the first and only reported studies to date for a complete, periodic lipopolysaccharide outer membrane. This also includes our current efforts in building on our initial and unique experience simulating the lipopolysaccharide membrane in the development and application of novel computational procedures and tools that allow molecular simulation studies of outer membrane proteins of Gram-negative bacteria to be carried out in realistic membrane models.

Straatsma, TP

2006-12-01

304

Effect of ionizing radiation on macrophage stimulating property of Vibrio parahaemolyticus lipopolysaccharide  

International Nuclear Information System (INIS)

Effect of gamma radiation on the macrophage stimulating ability of Vibrio parahaemolyticus lipopolysaccharide (LPS) was examined. Radiodetoxified LPS (RLPS) when injected (ip) in mice stimulated peritoneal macrophages as was evident from the enhancement of their acid hydrolases and cellular RNA contents. RLPS also stimulated the phagocytic activities of macrophages. The stimulation of macrophages was slightly less as compared to that observed with n ative LPS. Thus, treatment of LPS with 15 kGy dose of gamma radiation results in a slight reduction in its macrophage stimulating ability. (author). 3 tabs., 22 refs

305

Structural studies of the polysaccharides from the lipopolysaccharides of Azospirillum brasilense Sp246 and SpBr14.  

Science.gov (United States)

Lipopolysaccharides from closely related Azospirillum brasilense strains, Sp246 and SpBr14, were obtained by phenol-water extraction. Mild acid hydrolysis of the lipopolysaccharides followed by GPC on Sephadex G-50 resulted in polysaccharide mixtures. On the basis of sugar and methylation analyses, Smith degradation and (1)H and (13)C NMR spectroscopy data, it was concluded that both bacteria possess the same two distinct polysaccharides having structures 1 and 2: [structure: see text]. Structure 1 has been reported earlier for a polysaccharide of A. brasilense 54 [Fedonenko et al., 2011] whereas to our knowledge structure 2 has not been hitherto found in bacterial polysaccharides. PMID:25240180

Sigida, Elena N; Fedonenko, Yuliya P; Shashkov, Alexander S; Grinev, Vyacheslav S; Zdorovenko, Evelina L; Konnova, Svetlana A; Ignatov, Vladimir V; Knirel, Yuriy A

2014-10-29

306

Brilliant blue G attenuates lipopolysaccharide-mediated microglial activation and inflammation?  

Science.gov (United States)

Previous studies have confirmed that oxidized adenosine triphosphate, a P2X7 receptor antagonist, attenuates lipopolysaccharide-mediated microglial activation and inflammatory expression following neuronal damage in rat brain. NaCl and temperature may affect the potency of oxidized adenosine triphosphate. Brilliant blue G is a derivative of a widely used food additive and has little toxicity. This study explored the effects of brilliant blue G, a selective P2X7 receptor antagonist, on microglial activation and inflammation. Results demonstrated that brilliant blue G inhibited the release of cyclooxygenase-2 and interleukin-6 in BV2 cells. Immunofluorescence displayed that brilliant blue G could suppress lipopolysaccharide-induced microglial activation. This study used RNA interference to block P2X7 receptor expression and found that small interfering RNA also suppressed the release of cyclooxygenase-2 and interleukin-6 in BV2 cells. These results suggested that downregulation of the P2X7 receptor by brilliant blue G was involved in the inhibition of microglial activation and inflammation.

Lu, Kui; Wang, Jue; Hu, Bin; Shi, Xiaolei; Zhou, Junyi; Tang, Yamei; Peng, Ying

2013-01-01

307

Ragweed pollen extract intensifies lipopolysaccharide-induced priming of NLRP3 inflammasome in human macrophages.  

Science.gov (United States)

Ragweed pollen extract (RWE) possesses intrinsic NADPH oxidase activity that induces oxidative stress by initiating the production of intracellular reactive oxygen species (ROS). The ROS are important contributors to the manifestation of allergic inflammation; furthermore, concomitant exposure to an allergen and an endotoxin trigger a stronger inflammatory response. One of the main pro-inflammatory cytokines produced in inflammatory responses is interleukin-1? (IL-1?), and its production is associated with caspase-1-containing inflammasome complexes. Intracellular ROS have been implicated in NLRP3 inflammasome-mediated IL-1? production, therefore, we aimed to study whether RWE influences the function of NLRP3 inflammasome. Here we describe that, in the presence of NADPH, RWE significantly elevates lipopolysaccharide-induced IL-1? production of THP-1 cells as well as human primary macrophages and dendritic cells. We also demonstrate that increased IL-1? production is mediated through NLRP3 inflammasome in THP-1 macrophages. We provide evidence that RWE elevates cytosolic ROS level in these cells, and ROS inhibitors abolish IL-1? production. Furthermore, we show that RWE enhances lipopolysaccharide-induced gene transcription/expression of pro-IL-1? and key components of the inflammasome via a ROS-dependent mechanism. PMID:23278511

Varga, Aliz; Budai, Marietta M; Milesz, Sándor; Bácsi, Attila; T?zsér, József; Benk?, Szilvia

2013-04-01

308

Exogenous normal lymph alleviates lipopolysaccharide-induced acute lung injury through lessening the adhesion molecules  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english PURPOSE: To evaluate the role of exogenous normal lymph (ENL) on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in rats. METHODS: ALI was induced by the jugular vein injection of LPS (iv, 15 mg/kg) in rats of the LPS and LPS+ENL groups within 15 min, then, ENL without cell components [...] (5 ml/kg) was infused at the speed of 0.5 ml per minute in the LPS+ENL group, the same amount of saline was administered in the LPS group. The rats in the sham group received the same surgical procedure and saline. The histomorphology and the levels of P-selectin, intercellular adhesion molecule-1 (ICAM-1), myeloperoxidase (MPO) in pulmonary tissue were assessed. RESULTS: LPS induced pulmonary injury as well as increased the wet/dry weight ratio (W/D) and the levels of P-selectin, ICAM-1, and MPO in pulmonary tissues. These deleterious effects of LPS were significantly ameliorated by ENL treatment. CONCLUSION: Exogenous normal lymph could markedly alleviate the acute lung injury induced by lipopolysaccharide, and its effects might be related to lessening the adhesion molecules.

Li-li, Zhang; Zi-gang, Zhao; Chun-yu, Niu; Jing, Zhang.

2014-05-01

309

Exogenous normal lymph alleviates lipopolysaccharide-induced acute lung injury through lessening the adhesion molecules  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english PURPOSE: To evaluate the role of exogenous normal lymph (ENL) on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in rats. METHODS: ALI was induced by the jugular vein injection of LPS (iv, 15 mg/kg) in rats of the LPS and LPS+ENL groups within 15 min, then, ENL without cell components [...] (5 ml/kg) was infused at the speed of 0.5 ml per minute in the LPS+ENL group, the same amount of saline was administered in the LPS group. The rats in the sham group received the same surgical procedure and saline. The histomorphology and the levels of P-selectin, intercellular adhesion molecule-1 (ICAM-1), myeloperoxidase (MPO) in pulmonary tissue were assessed. RESULTS: LPS induced pulmonary injury as well as increased the wet/dry weight ratio (W/D) and the levels of P-selectin, ICAM-1, and MPO in pulmonary tissues. These deleterious effects of LPS were significantly ameliorated by ENL treatment. CONCLUSION: Exogenous normal lymph could markedly alleviate the acute lung injury induced by lipopolysaccharide, and its effects might be related to lessening the adhesion molecules.

Li-li, Zhang; Zi-gang, Zhao; Chun-yu, Niu; Jing, Zhang.

310

Activities of gemifloxacin (SB 265805, LB20304) compared to those of other oral antimicrobial agents against unusual anaerobes.  

Science.gov (United States)

The activities of gemifloxacin (SB 265805, LB20304) and comparator agents were determined by an agar dilution method against 419 clinical strains of less-commonly identified species of anaerobes. Gemifloxacin was generally more active than trovafloxacin against gram-positive strains by one to two dilutions. Peptostreptococci (Peptostreptococcus asaccharolyticus, Peptostreptococcus magnus, Peptostreptococcus micros, and Peptostreptococcus prevotii) and Porphyromonas spp. (Porphyromonas asaccharolytica, Porphyromonas canoris, Porphyromonas gingivalis, and Porphyromonas macacae) were all susceptible to Prevotella intermedia, Prevotella heparinolytica, and the Prevotella oris-buccae group. Fusobacterium naviforme and Fusobacterium necrophorum were also susceptible to Prevotella melaninogenica group, Prevotella bivia, Clostridium difficile, and Bilophila wadsworthia were relatively resistant to gemifloxacin (MIC(90)s, >/=4 microgram/ml). PMID:10543754

Goldstein, E J; Citron, D M; Vreni Merriam, C; Tyrrell, K; Warren, Y

1999-11-01

311

Effects of prepartal oronasal administration of lipopolysaccharide on milk composition and productivity of transition Holstein dairy cows  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The aim of this study was to evaluate the effects of repeated oronasal administration of lipopolysaccharide (LPS) on milk composition and the overall productive performance of dairy cows. One hundred pregnant Holstein dairy cows were randomly assigned to two treatment groups (n = 50). 30 cows out of 100 were selected for intensive sampling (n = 15) starting at 28 d before parturit...

Summera Iqbal; Qendrim Zebeli; Mansmann, Dominik A.; Dunn, Suzanna M.; Ametaj, Burim N.

2013-01-01

312

Lipopolysaccharides directly decrease Ca2+ oscillations and the hyperpolarization-activated nonselective cation current If in immortalized HL-1 cardiomyocytes.  

Science.gov (United States)

Lipopolysaccharide (LPS) has been implicated in sepsis-mediated heart failure and chronic cardiac myopathies. We determined that LPS directly and reversibly affects cardiac myocyte function by altering regulation of intracellular Ca2+ concentration ([Ca2+]i) in immortalized cardiomyocytes, HL-1 cells. [Ca2+]i oscillated (cardiomyocytes via TLR-4 and other mechanisms. PMID:20573997

Wondergem, Robert; Graves, Bridget M; Ozment-Skelton, Tammy R; Li, Chuanfu; Williams, David L

2010-09-01

313

Selenium Suppresses Lipopolysaccharide-Induced Fibrosis in Peritoneal Mesothelial Cells Through Inhibition of Epithelial-to-Mesenchymal Transition.  

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Peritoneal fibrosis resulting from long-term clinical peritoneal dialysis has been the main reason of dropout from peritoneal dialysis. Peritonitis as a common complication of peritoneal dialysis treatment may lead to the occurrences of peritoneal fibrosis. We cultured peritoneal mesothelial cells with lipopolysaccharides (LPS) in order to stimulate the environment of peritonitis and investigate whether lipopolysaccharides could induce epithelial-to-mesenchymal transition (EMT). Oxidative stress could stimulate fibrogenesis while selenium has antioxidant properties. So, this study also explored whether selenium supplementation affects lipopolysaccharide-induced EMT and fibrosis. We found that lipopolysaccharides could activate EMT changes such as the loss of E-cadherin and the increase of ?-smooth muscle actin (?-SMA), collagen I, vimentin, and fibronectin (FN), while selenium inhibits EMT by modulating reactive oxygen species (ROS) generation and ROS/MMP-9 signaling pathways in peritoneal mesothelial cells. Moreover, it was revealed that selenium decreased the EMT events of peritoneal mesothelial cells via inhibition of PI3k/AKT pathways. In conclusion, these findings enable a better understanding of the mechanism of peritoneal fibrosis and explore a new idea for the prevention and treatment. PMID:25108639

Liu, Jinyan; Zeng, Lingling; Zhao, Yuliang; Zhu, Bin; Ren, Wanjun; Wu, Chunling

2014-11-01

314

Cannabidiol reduces lipopolysaccharide-induced vascular changes and inflammation in the mouse brain: an intravital microscopy study  

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Abstract Background The phytocannabinoid cannabidiol (CBD) exhibits antioxidant and antiinflammatory properties. The present study was designed to explore its effects in a mouse model of sepsis-related encephalitis by intravenous administration of lipopolysaccharide (LPS). Methods Vascular responses of pial vessels were analyzed by intravital microscopy and inflammatory parameters measured by qRT-PCR. Results CBD prevented LPS-induced arteriolar an...

Tolón Rosa M; Millán África; Benito Cristina; Martínez-Orgado José A; Ruiz-Valdepeñas Lourdes; Romero Julia?n

2011-01-01

315

Oral Chlamydia trachomatis in Patients with Established Periodontitis  

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Periodontitis is considered a consequence of a pathogenic microbial infection at the periodontal site and host susceptibility factors. Periodontal research supports the association of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, and Bacteroides forsythus, and periodontitis; however causality has not been demonstrated. In pursuit of the etiology of periodontitis, we hypothesized that the intracellular bacteria, Chlamydia trachomatis, may play a role. A...

Reed, Susan G.; Lopatin, Dennis E.; Foxman, Betsy; Burt, Brian A.

2000-01-01

316

In vitro study of 980nm diode laser in dental implant disinfection  

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Full Text Available Objective: To evaluate the potential of 980nm diode laser to reduce bacteria after irradiation of three different dental implant surfaces contaminated with Enterococcus faecalis and Porphyromonas gingivalis, as well as the possible changes in the irradiated implant surfaces.Methods: Seventy two implants with machined surfaces, airborne particle abraded with titanium oxide and acid-etched surfaces were exposed to Enterococcus faecalis and Porphyromonas gingivalis cultures and irradiated with 980nm diode laser with power of 2.5 and 3,0W. After laser treatments, the number of remaining colony-forming units was studied and implant surface morphology was analyzed by scanning electron microscopy. Results: The results showed 100% reduction of the bacteria on the implants irradiated with 3.0W. Moreover, 100% reduction of bacteria was also achieved on the implant surfaces contaminated with Porphyromonas gingivalis when irradiated with 2.5W and 3.0W. Bacteria reduction was not complete for the implants contaminated with Enterococcus faecalis, irradiated with 2.5W and surfaces treated with TiO2 airborne particle abrasion (78.6% and acid etching (49.4%.The scanning electron microscopy analysis showed that at the power settings used, no implant surface changes were found. Conclusion: The 980nm diode laser was effective in decontaminating the Enterococcus faecalis and Porphyromonas gingivalis without promoting surface alteration in the implants.

Fábio Gonçalves

2009-12-01

317

In Vitro cytotoxicity and antibacterial activity of selected South African medicinal plants used in the treatment of periodontitis  

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The use of medicinal plants in the treatment of infectious diseases is an acceptable and popular phenomenon in South Africa and worldwide. The potential of extracts from these plants as antimicrobial agents necessitates their scientific evaluation. Therefore, this study evaluated the antimicrobial activity of Carpobrotus edulis; Cotyledon orbiculata; Datura stramonium; Dodonaea angustifolia; and Zanthoxylum capense against Porphyromonas gingivalis; Tannerella forsythensis and Actinobacillus a...

Maja, Matshidiso Patricia

2009-01-01

318

Identification of a methioninase inhibitor, myrsinoic acid B, from Myrsine seguinii Lév., and its inhibitory activities.  

Science.gov (United States)

A methioninase inhibitor from Myrsine seguinii was purified and identified as myrsinoic acid B. Its inhibitory activities as to crude methioninase from periodontal bacteria such as Fusobacterium nucleatum, Porphyromonas gingivalis, and Treponema denticola were determined. The IC50 values were 10.5, 82.4, and 30.3 microM respectively. PMID:18776663

Ito, Satomi; Narise, Atsushi; Shimura, Susumu

2008-09-01

319

Ellagic acid protects Lipopolysaccharide/d-galactosamine-induced acute hepatic injury in mice.  

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Ellagic acid, a natural polyphenol found in certain fruits, nuts and vegetables, has been reported to have anti-inflammatory, anti-tumor, and antioxidant activities. However, the effects of ellagic acid on acute hepatic injury have not been reported. In the present study, we investigated the effects of ellagic acid on Lipopolysaccharide/d-galactosamine-induced acute hepatic injury in mice. The results showed that LPS/GalN increased hepatic malondialdehyde (MDA) content, TNF-? level, serum ALT and AST levels and TNF-? level. However, these changes were attenuated by ellagic acid. Western blot analysis showed that ellagic acid inhibited LPS/GalN-induced NF-?B activation. Furthermore, ellagic acid induced the expression of Nrf2 and heme oxygenase-1. In conclusion, our results showed that ellagic acid protected against LPS/GalN-induced liver injury by enhancing the antioxidative defense system and reducing inflammatory response. PMID:25038320

Gu, Lei; Deng, Wen-Sheng; Liu, Ye; Jiang, Chun-Hui; Sun, Long-Ci; Sun, Xiao-Fei; Xu, Qing; Zhou, Hong

2014-10-01

320

HDAC6 Deacetylase Activity Is Critical for Lipopolysaccharide-Induced Activation of Macrophages  

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Activated macrophages play an important role in both innate and adaptive immune responses, and aberrant activation of macrophages often leads to inflammatory and immune disorders. However, the molecular mechanisms of how macrophages are activated are not fully understood. In this study, we identify a novel role for histone deacetylse 6 (HDAC6) in lipopolysaccharide (LPS)-induced macrophage activation. Our data show that suppression of HDAC6 activity significantly restrains LPS-induced activation of macrophages and production of pro-inflammatory cytokines. Further study reveals that the regulation of macrophage activation by HDAC6 is independent of F-actin polymerization and filopodium formation; instead, it is mediated by the effects of HDAC6 on cell adhesion and microtubule acetylation. These data thus suggest that HDAC6 is an important regulator of LPS-induced macrophage activation and might be a potential target for the management of inflammatory disorders.

Liu, Zhu; Ran, Jie; Li, Yuanyuan; Wang, Jian; Yang, Yang; Zhou, Jun; Li, Dengwen; Liu, Min

2014-01-01

 
 
 
 
321

Gram-Negative Marine Bacteria: Structural Features of Lipopolysaccharides and Their Relevance for Economically Important Diseases  

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Full Text Available Gram-negative marine bacteria can thrive in harsh oceanic conditions, partly because of the structural diversity of the cell wall and its components, particularly lipopolysaccharide (LPS. LPS is composed of three main parts, an O-antigen, lipid A, and a core region, all of which display immense structural variations among different bacterial species. These components not only provide cell integrity but also elicit an immune response in the host, which ranges from other marine organisms to humans. Toll-like receptor 4 and its homologs are the dedicated receptors that detect LPS and trigger the immune system to respond, often causing a wide variety of inflammatory diseases and even death. This review describes the structural organization of selected LPSes and their association with economically important diseases in marine organisms. In addition, the potential therapeutic use of LPS as an immune adjuvant in different diseases is highlighted.

Muhammad Ayaz Anwar

2014-04-01

322

Chemical and serological investigations on the genus-specific lipopolysaccharide epitope of Chlamydia.  

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Members of the bacterial genus Chlamydia are responsible for widespread disease among humans and animals, including endemic trachoma in developing countries, venereal disease in developed countries, and a variety of other diseases such as infantile pneumonia and lymphogranuloma venereum. Although there is little genetic relatedness between and large antigenic diversity between and among the two chlamydial species, one antigenic determinant has been preserved among all serovars: the genus-specific lipopolysaccharide epitope. In this report, the tools of molecular genetics, monoclonal antibodies, and analytical and synthetic chemistry have been combined to determine the structure of this epitope. This epitope is attributed to the presence of a trisaccharide of 3-deoxy-D-manno-octulosonic acid (KDO) of the sequence KDOp-(2----8)-KDOp-(2----4)-KDO. The structure includes a unique linkage of two KDO residues through a 2.8-linkage. PMID:2436232

Brade, H; Brade, L; Nano, F E

1987-01-01

323

Functional identification of Proteus mirabilis eptC gene encoding a core lipopolysaccharide phosphoethanolamine transferase.  

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By comparison of the Proteus mirabilis HI4320 genome with known lipopolysaccharide (LPS) phosphoethanolamine transferases, three putative candidates (PMI3040, PMI3576, and PMI3104) were identified. One of them, eptC (PMI3104) was able to modify the LPS of two defined non-polar core LPS mutants of Klebsiella pneumoniae that we use as surrogate substrates. Mass spectrometry and nuclear magnetic resonance showed that eptC directs the incorporation of phosphoethanolamine to the O-6 of L-glycero-D-mano-heptose II. The eptC gene is found in all the P. mirabilis strains analyzed in this study. Putative eptC homologues were found for only two additional genera of the Enterobacteriaceae family, Photobacterium and Providencia. The data obtained in this work supports the role of the eptC (PMI3104) product in the transfer of PEtN to the O-6 of L,D-HepII in P. mirabilis strains. PMID:24756091

Aquilini, Eleonora; Merino, Susana; Knirel, Yuriy A; Regué, Miguel; Tomás, Juan M

2014-01-01

324

Microglial ablation and lipopolysaccharide preconditioning affects pilocarpine-induced seizures in mice  

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Activated microglia have been associated with neurodegeneration in patients and in animal models of Temporal Lobe Epilepsy (TLE), however their precise functions as neurotoxic or neuroprotective is a topic of significant investigation. To explore this, we examined the effects of pilocarpine-induced seizures in transgenic mice where microglia/macrophages were conditionally ablated. We found that unilateral ablation of microglia from the dorsal hippocampus did not alter acute seizure sensitivity. However, when this procedure was coupled with lipopolysaccharide (LPS) preconditioning (1 mg/kg given 24 h prior to acute seizure), we observed a significant pro-convulsant phenomenon. This effect was associated with lower metabolic activation in the ipsilateral hippocampus during acute seizures, and could be attributed to activity in the mossy fiber pathway. These findings reveal that preconditioning with LPS 24 h prior to seizure induction may have a protective effect which is abolished by unilateral hippocampal microglia/macrophage ablation.

Mirrione, M.M.; Mirrione, M.M.; Konomosa, D.K.; Ioradanis, G.; Dewey, S.L.; Agzzid, A.; Heppnerd, F.L.; Tsirka, St.E.

2010-04-01

325

Influence of histamine in a liver injury model induced by Propionibacterium acnes and lipopolysaccharide.  

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In normal mice, plasma histamine levels were 29.4+/-10.1 pmol/ml. When 0.1 microg of lipopolysaccharide (LPS) was intravenously injected into Propionibacterium acnes (P. acnes)-primed ICR mice, histamine levels increased remarkably to 61.2+/-15.9 pmol/ml (pmice treated with 50 mg/kg of ranitidine, in contrast with 50% in the control group although the treatment did not significantly decrease the plasma ALT activity. On the other hand, 50 mg/kg of pyrilamine significantly reduced plasma ALT activity (p<0.001). The results suggested that histamine levels are related to hepatic damage in the P. acnes plus LPS induction of liver injury. PMID:14519942

Kurihara, Hiroshi; Fukami, Harukazu; Asami, Sumio; Shibata, Hiroshi; Kiso, Yoshinobu; Tanaka, Takaharu; Yao, Xin-Sheng

2003-10-01

326

Differential Stimulatory Activities of Smooth and Rough Brucella abortus Lipopolysaccharide in Murine Macrophages  

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Full Text Available Brucella abortus lipopolysaccharide (LPS was isolated and purified from rough (RB51 and smooth (S2308 strains of Brucella. The LPS preparations were used to treat murine (RAW 264.7 macrophages in order to study their differential effects. Treated macrophages were tested by lysozyme release test (LRT, nitroblue tetrazolium test (NBT and nitric oxide (NO assay, respectively. Rough Brucella LPS induced significantly higher levels of lysozyme release, oxidative stress, and nitric oxide in murine macrophages than smooth Brucella LPS or combined LPS (rough + smooth LPS. These responses were dose-dependent. Macrophages treated with rough LPS were more Brucellacidal than those treated with smooth LPS. The minimal stimulation of murine macrophages by Brucella smooth LPS may provide basis for less active immune responses against smooth strains.

Raheela Akhtar1,2*, Yongqun O. He2, Charles B. Larson2, Zafar I. Chaudhary3 and Mansur ud-Din Ahmad4

2012-06-01

327

The Lipopolysaccharide Export Pathway in Escherichia coli: Structure, Organization and Regulated Assembly of the Lpt Machinery  

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Full Text Available The bacterial outer membrane (OM is a peculiar biological structure with a unique composition that contributes significantly to the fitness of Gram-negative bacteria in hostile environments. OM components are all synthesized in the cytosol and must, then, be transported efficiently across three compartments to the cell surface. Lipopolysaccharide (LPS is a unique glycolipid that paves the outer leaflet of the OM. Transport of this complex molecule poses several problems to the cells due to its amphipatic nature. In this review, the multiprotein machinery devoted to LPS transport to the OM is discussed together with the challenges associated with this process and the solutions that cells have evolved to address the problem of LPS biogenesis.

Alessandra Polissi

2014-02-01

328

Cyclooxygenase-1 is involved in the inhibition of hippocampal neurogenesis after lipopolysaccharide-induced neuroinflammation.  

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Growing evidence indicates that neuroinflammation can alter adult neurogenesis by mechanisms as yet unclear. We have previously demonstrated that the neuroinflammatory response and neuronal damage after lipopolysaccharide (LPS) injection is reduced in cyclooxygenase-1 deficient (COX-1(-/-)) mice. In this study, we investigated the role of COX-1 on hippocampal neurogenesis during LPS-induced neuroinflammation, using COX-1(-/-) and wild type (WT) mice. We found that LPS-induced neuroinflammation resulted in the decrease of proliferation, survival and differentiation of hippocampal progenitor cells in WT but not in COX-1(-/-) mice. Thus, we demonstrate for the first time that COX-1 is involved in the inhibition of BrdU progenitor cells in proliferation and hippocampal neurogenesis after LPS. These results suggest that COX-1 may represent a viable therapeutic target to reduce neuroinflammation and promote neurogenesis in neurodegenerative diseases with a strong inflammatory component. PMID:21694498

Russo, Isabella; Amornphimoltham, Panomwat; Weigert, Roberto; Barlati, Sergio; Bosetti, Francesca

2011-08-01

329

Suppression of ConA-induced inflammatory ascites by lipopolysaccharide (LPS) in mice.  

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The effect of pre-treatment with Escherichia coli O83 lipopolysaccharide (LPS) on concanavalin A-induced ascites was examined. The LPS was injected intraperitoneally (i.p.) in different doses to mice, and then ascites was induced by i.p. administration of concanavalin A (ConA) (25 mg/kg b.w.). After 2.5 h the mice were killed and the ascitic fluid was collected and measured. The LPS produced a marked and dose-dependent inhibition of ConA-induced ascites and the effect of pre-treatment lasted up to almost a week. Complete inhibition could not be achieved. If administered alone, LPS did not produce ascites.It is well known that LPS enhances vascular permeability in several tissues, but the present work shows that peritoneal permeability is not enhanced by this agent. Suppression of ConA-induces ascites may be explained by the hypotonic effect of LPS. PMID:22982642

Baintner, Károly

2012-09-01

330

Interference of Salmonella typhimurium lipopolysaccharide on performance and biological parameters of broiler chickens  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english This study was conducted to determine the interference of Salmonella typhimurium lipopolysaccharide (sLPS) on the performance, biological parameters, and histological evaluations of 198 one-day-old male broiler chickens divided into three treatments according to sLPS dose (0, 250, or 500 µg/applicat [...] ion/bird) that was applied to the birds every other day, from 15 to 27 days of age. At the end of the experiment (28 days), significant effects were observed on body weight (R= -0.17 and P=0.05), total cholesterol serum levels(R=0.43 and p

RH, Rauber; VJ, Perlin; CD, Fin; AL, Mallmann; DP, Miranda; LZ, Giacomini; VP do, Nascimento.

331

Oenothein B Suppresses Lipopolysaccharide (LPS-Induced Inflammation in the Mouse Brain  

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Full Text Available Oenothein B has been recently evaluated for its ability to affect inflammatory responses in peripheral tissues. In this study, we examined its effect on the damage to the central nervous system due to systemic inflammation. For this purpose, ICR mice were injected with an intraperitoneal (i.p. dose of lipopolysaccharide (LPS; 1 mg/kg mouse. When oenothein B was administered per os (p.o., it suppressed (1 LPS-induced abnormal behavior in open field; (2 LPS-induced microglial activation in the hippocampus and striatum; and (3 LPS-induced cyclooxygenase (COX-2 production in the hippocampus and striatum of these mice. These results suggest that oenothein B had the ability to reduce neuroinflammation in the brain during systemic inflammation.

Yoshiko Furukawa

2013-05-01

332

Interference of Salmonella typhimurium lipopolysaccharide on performance and biological parameters of broiler chickens  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english This study was conducted to determine the interference of Salmonella typhimurium lipopolysaccharide (sLPS) on the performance, biological parameters, and histological evaluations of 198 one-day-old male broiler chickens divided into three treatments according to sLPS dose (0, 250, or 500 µg/applicat [...] ion/bird) that was applied to the birds every other day, from 15 to 27 days of age. At the end of the experiment (28 days), significant effects were observed on body weight (R= -0.17 and P=0.05), total cholesterol serum levels(R=0.43 and p

RH, Rauber; VJ, Perlin; CD, Fin; AL, Mallmann; DP, Miranda; LZ, Giacomini; VP do, Nascimento.

2014-03-01

333

Lipopolysaccharide (LPS)-binding proteins BPI and LBP form different types of complexes with LPS.  

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Lipopolysaccharide (LPS)-binding protein (LBP) and bactericidal/permeability-increasing protein (BPI) are closely related LPS-binding proteins whose binding to LPS has markedly different functional consequences. To gain better insight into the possible basis of these functional differences, the physical properties of LBP-LPS and BPI-LPS complexes have been compared in this study by sedimentation, light scattering, and fluorescence analyses. These studies reveal dramatic differences in the physical properties of LPS complexed to LBP versus BPI. They suggest that of the two proteins, only LBP can disperse LPS aggegates. However, BPI can enhance both the sedimentation velocity and apparent size of LPS aggregates while inhibiting LPS-LBP binding even at very low (1:40 to 1:20) BPI:LPS molar ratios. PMID:9228038

Tobias, P S; Soldau, K; Iovine, N M; Elsbach, P; Weiss, J

1997-07-25

334

Protective effect of carvacrol on acute lung injury induced by lipopolysaccharide in mice.  

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Carvacrol, the major component of Plectranthus amboinicus, has been known to exhibit anti-inflammatory activities. The aim of this study was to investigate the effects of carvacrol on lipopolysaccharide (LPS)-induced endotoxemia and acute lung injury (ALI) in mice. Mice were injected intraperitoneally (i.p.) with LPS and the mortality of mice for 7 days were observed twice a day. Meanwhile, the protective effect of carvacrol (20, 40 or 80 mg/kg) on LPS-induced endotoxemia were detected. Using an experimental model of LPS-induced ALI, we examined the effect of carvacrol in resolving lung injury. The results showed that carvacrol could improve survival during lethal endotoxemia and attenuate LPS-induced ALI in mice. The anti-inflammatory mechanisms of carvacrol may be due to its ability to inhibit NF-?B and MAPKs signaling pathways, thereby inhibiting inflammatory cytokines TNF-?, IL-6 and IL-1? production. PMID:24577726

Feng, Xiaosheng; Jia, Aiqing

2014-08-01

335

Antibody response to the lipopolysaccharide and protein antigens of Salmonella typhi during typhoid infection  

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Antibodies to the lipopolysaccharide (LPS) and protein antigens of S. typhi in secretions of small intestine obtained from 12 typhoid patients, four typhoid carriers and 16 non-typhoid control subjects were measured by a solid-phase radioimmunoassay technique using 125I labelled anti-immunoglobulin antibody. Intestinal secretions obtained from typhoid patients as a group had significantly higher anti-LPS and anti-protein antibodies than those from the control group. These antibodies were both IgM and IgA classes. There was no correlation between the IgM or IgA antibody levels in serum and those in the intestinal secretions. In the intestinal secretions obtained from typhoid carriers, on the other hand, only IgA-class antibodies to the LPS and protein antigens of S. typhi were present at high levels. (author)

336

Three different anti-lipopolysaccharide factors identified from giant freshwater prawn, Macrobrachium rosenbergii.  

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Anti-lipopolysaccharide factor (ALF) is a type of basic protein and an important antimicrobial peptide that can bind and neutralize lipopolysaccharides (LPS). This protein shows a broad spectrum of antimicrobial activity. In this study, three forms of ALF designated as MrALF5, MrALF6, and MrALF7 were identified from giant freshwater prawn, Macrobrachium rosenbergii. MrALF5, MrALF6, and MrALF7 genes encode 133, 121, and 120 amino acids of the corresponding proteins, respectively. All these ALF proteins contain LPS-binding domain with two conserved cysteine residues. The genomic sequences of MrALF5 and MrALF7 were amplified. The genomic structures of MrALF5 and MrALF7 comprise three exons interrupted by two introns. Phylogenetic analysis showed that MrALF5, MrALF6, and MrALF7 were clustered into clade II. Evolutionary analysis showed that ALF genes from M. rosenbergii may suffer a rapid evolution. MrALF5 was expressed mainly in the hepatopancreas, gills, and heart. MrALF6 was mainly distributed in the intestine and hepatopancreas. The highest expression level of MrALF7 was detected in the hepatopancreas. MrALF6, as well as MrALF7, was downregulated by Escherichia coli challenge, and all three ALF genes were upregulated by Vibrio or white spot syndrome virus challenge. MrALF6 was also upregulated by Staphylococcus aureus challenge. In summary, the three isoforms of ALF genes may participate in the innate immune response against bacteria and virus infecting the giant fresh water prawn. PMID:22800688

Ren, Qian; Zhang, Zhao; Li, Xin-Chang; Jie-Du; Hui, Kai-Min; Zhang, Chi-Yu; Wang, Wen

2012-10-01

337

Bacterial Lipopolysaccharides Pretreatment Protects Against Mutagenic and lmmunosuppressor Effects of Cyclophosphamide in Mice  

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Full Text Available The mutagenic and immunosuppressory effects of cyclophosphamide (CYP are still the primary limitation to wider application for treating a variety of human malignancies. On the one hand, CYP treatment predisposes transplant recipients and cancer patients to risk of bacterial, fungal, and viral infections, in the other word the, Lipopolysaccharide (LPS, an endotoxin found in cell walls of gram- negative bacteria, has been shown to play a significant role in development a of multiple Immune system responses and can cause a fatal pathological effect. The present investigation is focused on immunization of mice with the endotoxin LPS prior to CYP treatment, in an attempt to reduce mutagenicity and immunosuppressory effects caused by CYP. The in vivo anti-cytotoxicity and anti- mutagenicity of the inflammatory agents of bacterial Lipopolysaccharides (LPS isolated from Aeromonas hydrophila was evaluated by bone marrow chromosomal aberrations assay,differential white blood cells (WBCs count and respiratory burst enzymatic assays for phagocytosis in mice exposed to CYP. The data presented in this article indicates that, treatment with low dose of bacterial LPS once a week for four weeks at a dose of •," ml of LPS suspension (o• µg/kg mice/week, was non- cytotoxic and un-mutagenic to the animal cells. However, pretreatment with low dose of bacterial LPS significantly increase cellular resistance to the mutagenic and immunosuppressory effects of CYP. In conclusion, this immunization protocol suggests that immunization of mice by LPS prior to CYP treatment may induce a number of adaptive antimutagenic and immune response molecular mechanisms.

Abdella E

2008-11-01

338

Acute lipopolysaccharide exposure facilitates epileptiform activity via enhanced excitatory synaptic transmission and neuronal excitability in vitro  

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Full Text Available Fei Gao,1,2 Zhiqiang Liu,3 Wei Ren,3 Wen Jiang1 1Department of Neurology, Xijing Hospital, Fourth Military Medical University, Xi’an 710032, People’s Republic of China; 2Department of Neurology, First Affiliated Hospital of Xi’an Medical University, Xi’an 710077, People’s Republic of China; 3College of Life Sciences, Shaanxi Normal University, Xi’an 710062, People’s Republic of China Abstract: Growing evidence indicates brain inflammation has been involved in the genesis of seizures. However, the direct effect of acute inflammation on neuronal circuits is not well known. Lipopolysaccharide (LPS has been used extensively to stimulate brain inflammatory responses both in vivo and in vitro. Here, we observed the contribution of inflammation induced by 10 ?g/mL LPS to the excitability of neuronal circuits in acute hippocampal slices. When slices were incubated with LPS for 30 minutes, significant increased concentration of tumor necrosis factor ? and interleukin 1? were detected by enzyme-linked immunosorbent assay. In electrophysiological recordings, we found that frequency of epileptiform discharges and spikes per burst increased 30 minutes after LPS application. LPS enhanced evoked excitatory postsynaptic currents but did not modify evoked inhibitory postsynaptic currents. In addition, exposure to LPS enhanced the excitability of CA1 pyramidal neurons, as demonstrated by a decrease in rheobase and an increase in action potential frequency elicited by depolarizing current injection. Our observations suggest that acute inflammation induced by LPS facilitates epileptiform activity in vitro and that enhancement of excitatory synaptic transmission and neuronal excitability may contribute to this facilitation. These results may provide new clues for treating seizures associated with brain inflammatory disease. Keywords: lipopolysaccharide, hippocampus, inflammation, epileptiform activity, synaptic transmission, neuronal excitability

Gao F

2014-08-01

339

A Glycam-Based Force Field for Simulations of Lipopolysaccharide Membranes: Parametrization and Validation  

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Lipopolysaccharides (LPS) comprise the outermost layer of the Gram-negative bacteria cell envelope. Packed onto a lipid layer, the outer membrane displays remarkable physical?chemical differences compared to cell membranes. The carbohydrate-rich region confers a membrane asymmetry that underlies many biological processes such as endotoxicity, antibiotic resistance, and cell adhesion. Furthermore, unlike membrane proteins from other sources, integral outer-membrane proteins do not consist of transmembrane ? helices; instead they consist of antiparallel ?-barrels, which highlights the importance of the LPS membrane as a medium. In this work, we present an extension of the GLYCAM06 force field that has been specifically developed for LPS membranes using our Wolf2Pack program. This new set of parameters for lipopolysaccharide molecules expands the GLYCAM06 repertoire of monosaccharides to include phosphorylated N- and O-acetylglucosamine, 3-deoxy-D-manno-oct-2- ulosonic acid, L-glycero-D-manno-heptose and its O-carbamoylated variant, and N-alanine-D-galactosamine. A total of 1 µs of molecular dynamics simulations of the rough LPS membrane of Pseudomonas aeruginosa PA01 is used to showcase the added parameter set. The equilibration of the LPS membrane is shown to be signi!cantly slower compared to phospholipid membranes, on the order of 500 ns. It is further shown that water molecules penetrate the hydrocarbon region up to the terminal methyl groups, much deeper than commonly observed for phospholipid bilayers, and in agreement with neutron diffraction measurements. A comparison of simulated structural, dynamical, and electrostatic properties against corresponding experimentally available data shows that the present parameter set reproduces well the overall structure and the permeability of LPS membranes in the liquid-crystalline phase.

Kirschner, Karl N.; Lins, Roberto D.; Maass, Astrid; Soares, Thereza A.

2012-11-13

340

Structural analysis of lipopolysaccharide produced by Heddleston serovars 10, 11, 12 and 15 and the identification of a new Pasteurella multocida lipopolysaccharide outer core biosynthesis locus, L6.  

Science.gov (United States)

Pasteurella multocida is a Gram-negative bacterial pathogen classified into 16 serovars based on lipopolysaccharide (LPS) antigens. Previously, we have characterized the LPS outer core biosynthesis loci L1, L2, L3, L5 and L7, and have elucidated the full range of LPS structures associated with each. In this study, we have determined the LPS structures produced by the type strains representing the serovars 10, 11, 12 and 15 and characterized a new LPS outer core biosynthesis locus, L6, common to all. The L6 outer core biosynthesis locus shares significant synteny with the L3 locus but due to nucleotide divergence, gene duplication and gene redundancy, the L6 and L3 LPS outer cores are structurally distinct. Using LPS structural and genetic differences identified in each L6 strain, we have predicted a role for most of the L6 glycosyltransferases in LPS assembly. Importantly, we have identified two glycosyltransferases, GctD and GatB, that differ by one amino acid, A162T, but use different donor sugars [uridine diphosphate (UDP)-Glc and UDP-Gal, respectively]. The longest outer core oligosaccharide, produced by the serovar 12 type strain, contained a terminal region consisting of ?-Gal-(1,4)-?-GlcNAc-(1,3)-?-Gal-(1,4)-?-Glc that was identical in structure to the vertebrate glycosphingolipid, paragloboside. Mimicry of host glycosphingolipids has been observed previously in P. multocida strains belonging to L3 LPS genotype, which produce LPS similar in structure to the globo-series of glycosphingolipids. The expression of a paragloboside-like oligosaccharide on the LPS produced by the serovar 12 type strain indicates that strains belonging to the L6 LPS genotype may also engage in molecular mimicry. PMID:24740556

Harper, Marina; St Michael, Frank; John, Marietta; Steen, Jason; van Dorsten, Lieke; Parnas, Henrietta; Vinogradov, Evgeny; Adler, Ben; Cox, Andrew D; Boyce, John D

2014-07-01

 
 
 
 
341

Structural Properties of Lipopolysaccharides from Rickettsia typhi and Rickettsia prowazekii and Their Chemical Similarity to the Lipopolysaccharide from Proteus vulgaris OX19 Used in the Weil-Felix Test  

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The lipopolysaccharides (LPSs) isolated from typhus group (TG) rickettsiae Rickettsia typhi and Rickettsia prowazekii were characterized by chemical analysis and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by silver staining. LPSs from two species of TG rickettsiae contained glucose, 3-deoxy-d-manno-octulosonic acid, glucosamine, quinovosamine, phosphate, and fatty acids (?-hydroxylmyristic acid and heneicosanoic acid) but not heptose. The O-polysaccharides ...

Amano, Ken-ichi; Williams, Jim C.; Dasch, Gregory A.

1998-01-01

342

Comparative studies on the analysis of glycoproteins and lipopolysaccharides by the gel-based microchip and SDS-PAGE  

Science.gov (United States)

In order to determine time efficiency between the gel-based microchip (LabChip) and traditional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), glycoproteins and lipopolysaccharides were analyzed in this study. After 90 min of gel electrophoresis, glycoproteins (bovine serum albumin, lysozyme, ovalbumin, and apo-transferrin) and fluorescent lipopolysaccharides (LPS-O and LPS-S) under reducing conditions could be analyzed by SDS-PAGE, and it would take (including imaging and analyzing) more than 3 h. The same sample could also be assayed on a Bioanalyzer in combination with the LabChip, and it would only need 30 min from start to finish. The assay software automatically calculated the size and concentration of each separated peak and displayed the results in real time, thus eliminating time-consuming procedures such as imaging and analyzing. Compared to the traditional reducing SDS-PAGE, LabChip has a faster turnaround time. PMID:19693351

Hsieh, Jung-Feng; Chen, Shui-Tein

2007-01-01

343

Macrophage-mediated nanoparticle delivery to the periodontal lesions in established murine model via Pg-LPS induction  

DEFF Research Database (Denmark)

We established a murine periodontitis model by local injection of lipopolysaccharide of Porphyromonas gingivalis (Pg-LPS) into the gingival sulcus of mandibular left incisor four times with 48-h interval. The histological examination of the periodontal tissues demonstrated that significant loss of periodontal bone and ligaments was observed in the lesion side with abundant inflammatory cell infiltration. Two days after the last injection, Cy5-labelled siRNA/chitosan particles were injected intraperitoneally (ip). The chitosan/siRNA particles were taken up by peritoneal macrophages, which subsequently migrated to the inflamed gingival area evaluated by in vivo imaging. The localization of macrophages in the inflamed region was further confirmed by immunofluorescent staining. The present report demonstrates that intragingival injection of Pg-LPS can be used to create an experimental model of periodontal inflammation in mice and that recruitment of macrophages with chitosan/siRNA nanoparticles to the inflamed area opens the possibility of an RNAi-based therapeutic approach using chitosan as a carrier in periodontitis.

Ma, Zhiwei; Dagnaes-Hansen, Frederik

2014-01-01

344

Osteoinductive and anti-inflammatory effect of royal jelly on periodontal ligament cells.  

Science.gov (United States)

Royal jelly (RJ) has been reported to possess several physiological and pharmacological properties such as the ability to prevent osteoporosis in rats and anti-inflammatory effects. We hypothesized that RJ could have beneficial effects on the prevention or treatment of periodontal diseases, which are chronic inflammatory diseases caused by bacterial infection that result in resorption of the tooth-supporting bone. We assessed the effect of RJ on mineralization in mouse periodontal ligament cell clone 22 (MPDL22 cells), which are of an osteogenic and cementogenic lineage. The mRNA expression of osteopontin, osteocalcin and osterix, and mineralized nodule formation were significantly enhanced in RJ-treated MPDL22 cells. In addition, we investigated the effects of RJ on the production of inflammatory cytokines from MPDL22 cells stimulated with lipopolysaccharide (LPS) of Porphyromonas gingivalis, a periodontopathic bacterium. RJ suppressed LPS-induced interleukin-6 and CXC chemokine ligand 10 production from MPDL22 cells. Furthermore, RJ suppressed the expression of CD54 in MPDL22 cells: CD54 is the adhesion molecule involved in the accumulation of leukocytes in periodontal lesions. These findings suggest that the osteoinductive and anti-inflammatory effects of RJ can provide benefits for the treatment and prevention of periodontal diseases. PMID:21878736

Yanagita, Manabu; Kojima, Yuko; Mori, Kenta; Yamada, Satoru; Murakami, Shinya

2011-08-01

345

Synthesis and biological activities of 2,6-dihydroxy-4-isopentenyloxychalcone as an antimicrobial and anti-inflammatory compound.  

Science.gov (United States)

Chalcones are a group of plant-derived polyphenolic compounds possessing a wide variety of biological activities. The aim of this study was to synthesize 2,6-dihydroxy-4-isopentenyloxychalcone (1), a chalcone found in plants belonging to the genera Helichrysum, Pleiotaxix and Metalasia, and evaluate its antimicrobial effects against major oral pathogens as well as its anti-inflammatory properties. Compound 1 was synthesized using a simple two-step procedure. Among the seven pathogens tested, Porphyromonas gingivalis and Prevotella intermedia were the most susceptible to inhibition by 1. This chalcone also attenuated the secretion of interleukin-8 and chemokine (C-C motif) ligand 5 (CCL5) by lipopolysaccharide (LPS)-stimulated oral epithelial cells as well as the secretion of matrix metalloproteinase 2 (MMP-2) by LPS-stimulated gingival fibroblasts. In conclusion, our study showed that 1 exerts a dual effect by acting on both oral pathogens and the host inflammatory response and thus represents a molecule of interest for periodontal infections. PMID:23859000

Bonifait, Laetitia; Zhao, Lei; Azelmat, Jabrane; Genovese, Salvatore; Epifano, Francesco; Grenier, Daniel

2014-05-01

346

Lipopolysaccharide challenge-induced suppression of Fos in hypothalamic orexin neurons: their potential role in sickness behavior  

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Orexin neurons in the lateral hypothalamus constitute a critical component in regulation of waking, feeding, and reward-related behaviors. In this study we examined the effects of lipopolysaccharide (LPS) challenge on Fos expression in orexin neurons in rats, to determine changes during sickness in two different behavioral contexts. One cohort of rats was treated with saline or LPS during the daytime, and then tested on an elevated plus maze (EPM) or left in their home cage until sacrifice. A...

Gaykema, Ronald P. A.; Goehler, Lisa E.

2009-01-01

347

Cytokine induction by lipopolysaccharide (LPS) corresponds to lethal toxicity and is inhibited by nontoxic Rhodobacter capsulatus LPS.  

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Many pathological effects of gram-negative bacteria are produced by their cell wall-derived lipopolysaccharides (LPSs). Differing pathogenicity of gram-negative LPSs, however, may depend on their capacities to induce cytokines. Thus, we studied the lethal toxicity of four nonenterobacterial LPSs and compared it with their capacity to induce mononuclear cell (MNC)-derived interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor (TNF). Unstimulated MNC did not release these cytokin...

Loppnow, H.; Libby, P.; Freudenberg, M.; Krauss, J. H.; Weckesser, J.; Mayer, H.

1990-01-01

348

Priming of human monocytes for enhanced lipopolysaccharide responses: expression of alpha interferon, interferon regulatory factors, and tumor necrosis factor.  

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Culture of human monocytes with either granulocyte-macrophage colony-stimulating factor or gamma interferon (IFN-gamma) results in a primed state, during which these cells express heightened responses to bacterial lipopolysaccharide (LPS). The production of IFN-alpha in response to LPS by human monocytes has an absolute requirement for priming. Tumor necrosis factor (TNF) expression is also greatly enhanced in primed monocytes after LPS stimulation, but unlike IFN-alpha, TNF is readily expres...

Hayes, M. P.; Zoon, K. C.

1993-01-01

349

Antibiotic-Induced Lipopolysaccharide (LPS) Release from Salmonella typhi: Delay between Killing by Ceftazidime and Imipenem and Release of LPS  

Digital Repository Infrastructure Vision for European Research (DRIVER)

It has been suggested that the antibiotic-induced release of lipopolysaccharide (LPS) is an important cause of the development of septic shock in patients treated for severe infections caused by gram-negative bacteria. ?-Lactam antibiotics change the integrity of the bacterial cell envelope by binding to penicillin-binding proteins (PBP) in the membrane and thus may affect the amount of LPS that is released and the kinetics of that release. In this respect, ceftazidime at intermediate concen...

Langevelde, Petra; Kwappenberg, Kitty M. C.; Groeneveld, Paul H. P.; Mattie, Herman; Dissel, Jaap T.

1998-01-01

350

Convergent synthesis of the tetrasaccharide repeating unit of the cell wall lipopolysaccharide of Escherichia coli O40  

Directory of Open Access Journals (Sweden)

Full Text Available A tetrasaccharide repeating unit corresponding to the cell-wall lipopolysaccharide of E. coli O40 was synthesized by using a convergent block glycosylation strategy. A disaccharide donor was coupled to a disaccharide acceptor by a stereoselective glycosylation. A 2-aminoethyl linker was chosen as the anomeric protecting group at the reducing end of the tetrasaccharide. All glycosylation steps are significantly high yielding and stereoselective.

Abhijit Sau

2012-11-01

351

Effects of Prenatal Lipopolysaccharide Exposure on Reproductive Activities and Serum Concentrations of Pituitary-Gonadal Hormones in Mice Offspring  

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Background: Maternal infection during pregnancy is a risk factor for some behavioralproblems with neurodevelopmental origin. This study aimed to evaluate the effects ofexposure of pregnant mice to the bacterial lipopolysaccharide (LPS) on sexual behaviourand serum level of pituitary-gonadal hormones of offspring in adulthood.Materials and Methods: In this Expremental study, pregnant NMRI mice (n=7/group)were treated with intra-peritoneal administration of LPS (1, 5 and 10 ?g/kg) at day 10of ...

Jalal Solati; Ramin Hajikhani; Behnam Rashidieh; Mahshid Fatipour Jalilian

2012-01-01

352

Bacterial-lipopolysaccharide-induced release of lactoferrin from human polymorphonuclear leukocytes: role of monocyte-derived tumor necrosis factor alpha.  

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We have examined the role played by human peripheral blood monocytes in mediating responses of human polymorphonuclear leukocytes (PMN) to bacterial lipopolysaccharide (LPS) in vitro. When incubated with Salmonella typhimurium LPS at 37 degrees C, human PMN suspended in serum-free buffer released the specific granule constituent lactoferrin into the surrounding medium. Release of lactoferrin from PMN varied with the concentration of LPS (1 to 1,000 ng/ml) as well as with the duration of incub...

Koivuranta-vaara, P.; Banda, D.; Goldstein, I. M.

1987-01-01

353

Lactoferrin Inhibits the Lipopolysaccharide-Induced Expression and Proteoglycan-Binding Ability of Interleukin-8 in Human Endothelial Cells  

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Interleukin-8 (IL-8), a C-X-C chemokine bound to endothelium proteoglycans, initiates the activation and selective recruitment of leukocytes at inflammatory foci. We demonstrate that human lactoferrin, an antimicrobial lipopolysaccharide (LPS)-binding protein, decreases both IL-8 mRNA and protein expression induced by the complex Escherichia coli 055:B5 LPS/sCD14 in human umbilical vein endothelial cells. The use of recombinant lactoferrins mutated in the LPS-binding sites indicates that this...

Elass, Elisabeth; Masson, Maryse; Mazurier, Joe?l; Legrand, Dominique

2002-01-01

354

Induction of C3b-mediated phagocytosis in macrophages by distinct populations of lipopolysaccharide-stimulated lymphocytes.  

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Unelicited resident peritoneal macrophages do not significantly ingest erythrocytes coated with C3b. However, these resident macrophages can be induced to ingest via the C3b receptor when cocultured with peritoneal or splenic nonadherent cells obtained from mice previously injected with lipopolysaccharide. In this study, the extent of ingestion induced in resident macrophages was dependent on the number of stimulated nonadherent cells cocultured with the macrophages as well as on the amount o...

Wrigley, D. M.; Saluk, P. H.

1981-01-01

355

Unusual Outer Membrane Lipid Composition of the Gram-negative, Lipopolysaccharide-lacking Myxobacterium Sorangium cellulosum So ce56*  

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The Gram-negative myxobacterium Sorangium cellulosum So ce56 bears the largest bacterial genome published so far, coding for nearly 10,000 genes. Careful analysis of this genome data revealed that part of the genes coding for the very well conserved biosynthesis of lipopolysaccharides (LPS) are missing in this microbe. Biochemical analysis gave no evidence for the presence of LPS in the membranes of So ce56. By analyzing the lipid composition of its outer membrane sphingolipids were identifie...

Keck, Matthias; Gisch, Nicolas; Moll, Hermann; Vorho?lter, Frank-jo?rg; Gerth, Klaus; Kahmann, Uwe; Lissel, Manfred; Lindner, Buko; Niehaus, Karsten; Holst, Otto

2011-01-01

356

Evaluation of IgM ELISA using a sonicate and a lipopolysaccharide antigen for the serodiagnosis of melioidosis  

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Melioidosis, caused by Burkholderia pseudomallei, has variable manifestations. The disease can present as an acute or a chronic form or localized or disseminated or can remain latent for many years. Acute septicaemic melioidosis has a high fatality rate when untreated and therefore, an early diagnosis is critical. Lack of testing facilities and of an awareness of the manifestations of the disease makes it likely that it is underreported in India. A sonicate and a lipopolysaccharide (LP...

Anandan S; Augustine A; Mathai E; Jesudason M

2010-01-01

357

Enzymatic Activities of Bovine Peripheral Blood Leukocytes and Milk Polymorphonuclear Neutrophils during Intramammary Inflammation Caused by Lipopolysaccharide  

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Leukocytes are recruited from peripheral blood into milk as part of the inflammatory response to mastitis. However, excessive accumulation of inflammatory cells alters the quality of milk and the proteases produced by polymorphonuclear neutrophils (PMNs) and macrophages may lead to mammary tissue damage. To investigate PMN recruitment and the kinetics of their intracytoplasmic enzymes in inflammation, we generated mastitis in six cows by intramammary infusion of lipopolysaccharide (LPS). Clin...

Prin-mathieu, C.; Le Roux, Y.; Laurent, F.; Be?ne?, M. C.; Moussaoui, F.

2002-01-01

358

Molecular evidence for the expression of angiotensin converting enzyme in hemocytes of Locusta migratoria: stimulation by bacterial lipopolysaccharide challenge.  

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The presence of angiotensin converting enzyme (ACE) in insects has been reported many times, but numerous questions about the functional role of this enzyme in insects remain. Here we show by RT-PCR experiments that ACE has a wide tissue distribution in Locusta migratoria, suggesting diverse roles for this enzyme in the locust. Immune challenge through injection of bacterial lipopolysaccharides resulted in a tenfold increase of ACE gene transcripts in the hemocytes and is suggestive for a rol...

Macours, N.; Hens, K.; Francis, C.; Loof, A.; Huybrechts, R.

2003-01-01

359

Inhibitory Effect of Astragalus Polysaccharides on Lipopolysaccharide-Induced TNF-a and IL-1? Production in THP-1 Cells  

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Astragalus polysaccharides (APS), one of main bioactive components in Astragalus membranaceus Bunge, has been reported to possess anti-inflammatory activities, but the molecular mechanisms behind this activity are largely unknown. This study aimed to investigate expression of inflammatory cytokines and the MAPK/NF-?B pathway in human THP-1 macrophages induced by lipopolysaccharide (LPS). The results showed that the concentrations of TNF-a and IL-1? released from LPS stimulated THP-1 cells i...

Aiping Lu; Cheng Lu; Li Xu; Jun Shu; Xiaojuan He

2012-01-01

360

Regulation of Adenosine Receptor Subtypes during Cultivation of Human Monocytes: Role of Receptors in Preventing Lipopolysaccharide-Triggered Respiratory Burst  

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Adenosine is a potent anti-inflammatory agent that modulates the function of cells involved in the inflammatory response. Here we show that it inhibits lipopolysaccharide (LPS)-induced formation of reactive oxygen intermediates (ROI) in both freshly isolated and cultured human monocytes. Blocking of adenosine uptake and inactivation of the adenosine-degrading enzyme adenosine deaminase enhanced the inhibitory action of adenosine, indicating that both pathways regulate the extracellular adenos...

Thiele, Andrea; Kronstein, Romy; Wetzel, Anne; Gerth, Anja; Nieber, Karen; Hauschildt, Sunna

2004-01-01