WorldWideScience
1

Reducing the bioactivity of Tannerella forsythia lipopolysaccharide by Porphyromonas gingivalis.  

Science.gov (United States)

Tannerella forsythia is considered a pathogen of periodontitis and forms a biofilm with multi-species bacteria in oral cavity. Lipopolysaccharide is a powerful immunostimulator and induces inflammation and shock. The purpose of this study was to investigate the characteristics of T. forsythia LPS in its co-cultivation with Fusobacterium nucleatum or Porphyromonas gingivalis. T. forsythia was co-cultured in the presence and absence of F. nucleatum and P. gingivalis and then T. forsythia LPS was extracted. The extracts were analyzed by SDS-PAGE and NF-?B reporter CHO cell lines. THP-1 cells were treated with the LPS and evaluated induction of cytokine expression by real-time RT-PCR and ELISA. For analysis of the bioactivity of T. forsythia LPS, the binding assay on LPS-binding protein (LBP) and CD14 was processed. The extracts did not contaminate other molecules except LPS and showed TLR4 agonists. Co-cultured T. forsythia LPS with P. gingivalis exhibited a lower level of induction of TNF-?, IL-1?, and IL-6 expression than single- or co-cultured T. forsythia LPS with F. nucleatum in the conditions of human serum. However, the three T. forsythia LPS did not show difference of cytokine induction in the serum free conditions. Co-cultured T. forsythia LPS with P. gingivalis exhibited a lower affinity to LBP and CD14 as binding site of O-antigen and attached at a lower level to THP-1 cells compared to single- or co-cultured T. forsythia LPS with F. nucleatum. The virulence of T. forsythia LPS was decreased by co-culturing with P. gingivalis and their affinity to LBP and CD14 was reduced, which may due to modification of O-antigen chain by P. gingivalis. PMID:25098564

Kim, Young-Jae; Lee, Sung-Hoon

2014-08-01

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

OpenAIRE

To characterize the roles of Porphyromonas gingivalis and its components in the disease processes, we investigated the cytokine profile induced by live P. gingivalis, its lipopolysaccharides (LPS), and its major fimbrial protein, fimbrillin (FimA). Using cytokine antibody arrays, we found that P. gingivalis LPS and FimA induced a similar profile of cytokine expression when exposed to mouse peritoneal macrophages but that this profile differed significantly in response to live P. gingivalis. I...

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

2005-01-01

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Xylitol Inhibits Inflammatory Cytokine Expression Induced by Lipopolysaccharide from Porphyromonas gingivalis  

OpenAIRE

Porphyromonas gingivalis is one of the suspected periodontopathic bacteria. The lipopolysaccharide (LPS) of P. gingivalis is a key factor in the development of periodontitis. Inflammatory cytokines play important roles in the gingival tissue destruction that is a characteristic of periodontitis. Macrophages are prominent at chronic inflammatory sites and are considered to contribute to the pathogenesis of periodontitis. Xylitol stands out and is widely believed to possess anticaries propertie...

Han, Su-ji; Jeong, So-yeon; Nam, Yun-ju; Yang, Kyu-ho; Lim, Hoi-soon; Chung, Jin

2005-01-01

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Porphyromonas gingivalis lipopolysaccharide is poorly recognized by molecular components of innate host defense in a mouse model of early inflammation.  

OpenAIRE

Porphyromonas gingivalis is a gram-negative bacterium that is associated with periodontitis. It has been hypothesized that destruction of bone and periodontal connective tissue is associated with colonization of the subgingival crevicular space by P. gingivalis, although how these bacteria overcome innate host defenses is largely unknown. To examine the early cellular and molecular events of P. gingivalis interaction with host tissues, we compared lipopolysaccharide (LPS) isolated from this b...

Reife, R. A.; Shapiro, R. A.; Bamber, B. A.; Berry, K. K.; Mick, G. E.; Darveau, R. P.

1995-01-01

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Thrombospondin-1 Production Is Enhanced by Porphyromonas gingivalis Lipopolysaccharide in THP-1 Cells  

Science.gov (United States)

Periodontitis is a chronic inflammatory disease caused by gram-negative anaerobic bacteria. Monocytes and macrophages stimulated by periodontopathic bacteria induce inflammatory mediators that cause tooth-supporting structure destruction and alveolar bone resorption. In this study, using a DNA microarray, we identified the enhanced gene expression of thrombospondin-1 (TSP-1) in human monocytic cells stimulated by Porphyromonas gingivalis lipopolysaccharide (LPS). TSP-1 is a multifunctional extracellular matrix protein that is upregulated during the inflammatory process. Recent studies have suggested that TSP-1 is associated with rheumatoid arthritis, diabetes mellitus, and osteoclastogenesis. TSP-1 is secreted from neutrophils, monocytes, and macrophages, which mediate immune responses at inflammatory regions. However, TSP-1 expression in periodontitis and the mechanisms underlying TSP-1 expression in human monocytic cells remain unknown. Here using real-time RT-PCR, we demonstrated that TSP-1 mRNA expression level was significantly upregulated in inflamed periodontitis gingival tissues and in P. gingivalis LPS-stimulated human monocytic cell line THP-1 cells. TSP-1 was expressed via Toll-like receptor (TLR) 2 and TLR4 pathways. In P. gingivalis LPS stimulation, TSP-1 expression was dependent upon TLR2 through the activation of NF-?B signaling. Furthermore, IL-17F synergistically enhanced P. gingivalis LPS-induced TSP-1 production. These results suggest that modulation of TSP-1 expression by P. gingivalis plays an important role in the progression and chronicity of periodontitis. It may also contribute a new target molecule for periodontal therapy. PMID:25501558

Gokyu, Misa; Kobayashi, Hiroaki; Nanbara, Hiromi; Sudo, Takeaki; Ikeda, Yuichi; Suda, Tomonari; Izumi, Yuichi

2014-01-01

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

OpenAIRE

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  

Directory of Open Access Journals (Sweden)

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

DIEGO GUALTERO

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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 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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Expression of Toll-Like Receptor 2 in Glomerular Endothelial Cells and Promotion of Diabetic Nephropathy by Porphyromonas gingivalis Lipopolysaccharide  

OpenAIRE

The toll-like receptor (TLR) has been suggested as a candidate cause for diabetic nephropathy. Recently, we have reported the TLR4 expression in diabetic mouse glomerular endothelium. The study here investigates the effects of the periodontal pathogen Porphyromonas gingivalis lipopolysaccharide (LPS) which is a ligand for TLR2 and TLR4 in diabetic nephropathy. In laser-scanning microscopy of glomeruli of streptozotocin- and a high fat diet feed-induced type I and type II diabetic mice, TLR2 l...

Sawa, Yoshihiko; Takata, Shunsuke; Hatakeyama, Yuji; Ishikawa, Hiroyuki; Tsuruga, Eichi

2014-01-01

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

Science.gov (United States)

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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Porphyromonas gingivalis Lipopolysaccharide Upregulates Insulin Secretion From Pancreatic ? Cell Line MIN6  

Science.gov (United States)

Background A close association between periodontitis and diabetes has been demonstrated in human cross-sectional studies, but an exact relationship between periodontitis and prediabetes has not been established. Previous studies using animal model systems consistently have shown that hyperinsulinemia occurs in animals with periodontitis compared to animals with healthy periodontium (while maintaining normoglycemia). Because bacterial lipopolysaccharide (LPS) plays an important role in the pathogenesis of periodontitis, we hypothesized that LPS may stimulate insulin secretion through a direct effect on ? cell function. To test this hypothesis, pancreatic ? cell line MIN6 cells were used to determine the effect of Porphyromonas gingivalis (Pg) LPS on insulin secretion. Furthermore, expression of genes altered by Pg LPS in innate immunity and insulin-signaling pathways was determined. Methods MIN6 cells were grown in medium with glucose concentration of normoglycemia (5.5 mM). Pg LPS was added to each well at final concentrations of 50, 200, and 500 ng/mL. Insulin secretion was measured using enzyme-linked immunosorbent assay. Gene expression levels altered by Pg LPS were determined by polymerase chain reaction (PCR) array for mouse innate and adaptive immunity response and mouse insulin-signaling pathways, and results were confirmed for specific genes of interest by quantitative PCR. Results Pg LPS stimulated insulin secretion in the normoglycemic condition by ?1.5- to 3.0-fold depending on the concentration of LPS. Pg LPS treatment altered the expression of several genes involved in innate and adaptive immune response and insulin-signaling pathway. Pg LPS upregulated the expression of the immune response–related genes cluster of differentiation 8a (Cd8a), Cd14, and intercellular adhesion molecule-1 (Icam1) by about two-fold. LPS also increased the expression of two insulin signaling–related genes, glucose-6-phosphatase catalytic subunit (G6pc) and insulin-like 3 (Insl3), by three- to four-fold. Conclusions We have demonstrated for the first time that Pg LPS stimulates insulin secretion by pancreatic ? cell line MIN cells. Pg LPS may have significant implications on the development of ? cell compensation and insulin resistance in prediabetes in individuals with periodontitis. PMID:24921432

Bhat, Uppoor G.; Ilievski, Vladimir; Unterman, Terry G.; Watanabe, Keiko

2015-01-01

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Porphyromonas gingivalis fimbriae.  

Science.gov (United States)

Marginal periodontitis is not a homogeneous disease but is rather influenced by an intricate set of host susceptibility differences as well as diversities in virulence among the harbored organisms. It is likely that clonal heterogeneity of subpopulations with both high and low levels of pathogenicity exists among organisms harbored by individuals with negligible, slight, or even severe periodontal destruction. Therefore, specific virulent clones of periodontal pathogens may cause advanced and/or aggressive periodontitis. Porphyromonas gingivalis is a predominant periodontal pathogen that expresses a number of potential virulence factors involved in the pathogenesis of periodontitis, and accumulated evidence shows that its expression of heterogenic virulence properties is dependent on clonal diversity. Fimbriae are considered to be critical factors that mediate bacterial interactions with and invasion of host tissues, with P. gingivalis shown to express two distinct fimbria-molecules, long and short fimbriae, on the cell surface, both of which seem to be involved in development of periodontitis. Long fimbriae are classified into six types (I to V and Ib) based on the diversity of fimA genes encoding FimA (a subunit of long fimbriae). Studies of clones with type II fimA have revealed their significantly greater adhesive and invasive capabilities as compared to other fimA type clones. Long and short fimbriae induce various cytokine expressions such as IL-1?, IL-?, IL-6, and TNF-?, which result in alveolar bone resorption. Although the clonal diversity of short fimbriae is unclear, distinct short fimbria-molecules have been found in different strains. These fimbriae variations likely influence the development of periodontal disease. PMID:23667717

Enersen, Morten; Nakano, Kazuhiko; Amano, Atsuo

2013-01-01

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Porphyromonas gingivalis: a clonal pathogen?  

OpenAIRE

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 dive...

Morten Enersen

2011-01-01

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Porphyromonas gingivalis: a clonal pathogen?  

Directory of Open Access Journals (Sweden)

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

Morten Enersen

2011-11-01

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Dental follicle progenitor cells responses to Porphyromonas gingivalis LPS  

OpenAIRE

Periodontitis is a bacterially induced chronic inflammatory disease. Dental follicle progenitor cells (DFPCs) have been proposed as biological graft for periodontal regenerative therapies. The potential impact of bacterial toxins on DFPCs properties is still poorly understood. The aim of this study was to investigate whether DFPCs are able to sense and respond to lipopolysaccharide (LPS) from Porphyromonas gingivalis, a major periopathogenic bacterium. Specifically, we hypothesized that LPS c...

Chatzivasileiou, Kyriaki; Lux, Cornelia A.; Steinhoff, Gustav; Lang, Hermann

2013-01-01

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Porphyromonas gingivalis Strain Variability and Periodontitis  

OpenAIRE

To determine if there is variability in virulence among strains of Porphyromonas gingivalis in human periodontitis, their distribution in a group of subjects with clear indicators of periodontitis and in a healthy, age-matched control group was examined. The presence of heteroduplex types of P. gingivalis in the two groups was determined with a PCR-based assay. This assay relied on detection of polymorphisms in the ribosomal internal spacer region (ISR). ISR fragments generated by PCR with P....

Griffen, Ann L.; Lyons, Sharon R.; Becker, Mitzi R.; Moeschberger, Melvin L.; Leys, Eugene J.

1999-01-01

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Effect of irradiation on the Porphyromonas gingivalis  

International Nuclear Information System (INIS)

The aim of this study was to observe a direct effect of irradiation on the periodontopathic Porphyromonas gingivalis (P. gingivalis). P. gingivalis 2561 was exposed to irradiation with a single absorbed dose of 10, 20, 30, and 40 Gy. Changes in viability and antibiotic sensitivity, morphology, transcription, and protein profile of the bacterium after irradiation were examined by pour plating method, disc diffusion method, transmission electron microscopy, RT-PCR, and immunoblot, respectively. Viability of irradiated P. gingivalis drastically reduced as irradiation dose was increased. Irradiated P. gingivalis was found to have become more sensitive to antibiotics as radiation dose was increased. With observation under the transmission electron microscope, the number of morphologically abnormal cells was increased with increasing of irradiation dose. In RT-PCR, decrease in the expression of fim A and sod was observed in irradiated P. gingivalis. In immunoblot, change of profile in irradiated P. gingivalis was found in a number of proteins including 43-kDa fimbrillin. These results suggest that irradiation may affect the cell integrity of P. gingivalis, which is manifested by the change in cell morphology and antibiotic sensitivity, affecting viability of the bacterium.

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Porphyromonas gingivalis: Major Periodontopathic Pathogen Overview  

OpenAIRE

Porphyromonas gingivalis is a Gram-negative oral anaerobe that is involved in the pathogenesis of periodontitis and is a member of more than 500 bacterial species that live in the oral cavity. 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: this is attributed to its arsenal of specialized virulence factors. The purpose of this review is to provide an overv...

Mysak, Jaroslav; Podzimek, Stepan; Sommerova, Pavla; Lyuya-mi, Yelena; Bartova, Jirina; Janatova, Tatjana; Prochazkova, Jarmila; Duskova, Jana

2014-01-01

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Biochemical characterization of Porphyromonas (Bacteroides) gingivalis collagenase.  

OpenAIRE

A protease was purified from Porphyromonas gingivalis 1101, a clinical isolate, by sequential sodium dodecyl sulfate-polyacrylamide gel electrophoresis, substrate diffusion gel electrophoresis, and electroelution. The enzyme cleaved radiolabeled human basement membrane type IV collagen and the synthetic collagen peptide substrate for eukaryotic collagenases. It was inactivated by the thiol protease inhibitor N-ethylmaleimide but not by EDTA or EGTA [ethylene glycol-bis(beta-aminoethyl ether)-...

Lawson, D. A.; Meyer, T. F.

1992-01-01

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Sialidase and Sialoglycoproteases Can Modulate Virulence in Porphyromonas gingivalis ? †  

OpenAIRE

The Porphyromonas gingivalis recombinant VimA can interact with the gingipains and several other proteins, including a sialidase. Sialylation can be involved in protein maturation; however, its role in virulence regulation in P. gingivalis is unknown. The three sialidase-related proteins in P. gingivalis showed the characteristic sialidase Asp signature motif (SXDXGXTW) and other unique domains. To evaluate the roles of the associated genes, randomly chosen P. gingivalis isogenic mutants crea...

Aruni, Wilson; Vanterpool, Elaine; Osbourne, Devon; Roy, Francis; Muthiah, Arun; Dou, Yuetan; Fletcher, Hansel M.

2011-01-01

22

Inhibitory effects of Ginkgo biloba extract on inflammatory mediator production by Porphyromonas gingivalis lipopolysaccharide in murine macrophages via Nrf-2 mediated heme oxygenase-1 signaling pathways.  

Science.gov (United States)

Periodontitis is an oral chronic inflammatory disease that influences systemic diseases. Heme oxygenase-1 has several beneficial abilities through Nrf-2 regulation. Ginkgo biloba has been reported to have anti-inflammatory effects associated with heme oxygenase-1 (HO-1) expression. In this study, we investigated whether the anti-inflammatory effects of G. biloba were involved with Nrf-2-mediated HO-1 expression in Porphyromonas gingivalis LPS-stimulated RAW264.7 macrophage cells. G. biloba was extracted with ethyl acetate (EGB). EGB exhibited anti-inflammatory activities, which suppressed the production of pro-inflammatory mediators, the activation of mitogen-activated protein kinases, and nuclear translocation of transcription factors. EGB also up-regulated the HO-1 expression, and the Nrf-2 level in the nucleus and its transactivity. Furthermore, reduced pro-inflammatory mediator levels by EGB were inverted in the presence of SnPP. The collective results suggest that the anti-inflammatory effects of EGB are due to the HO-1 expression via up-regulation of Nrf-2 in RAW 264.7 cells stimulated by P. gingivalis LPS. PMID:22476936

Ryu, Eun Yeon; Park, Ah Jeong; Park, Sun Young; Park, Sung Hae; Eom, Hye Won; Kim, Young Hun; Park, Geuntae; Lee, Sang-Joon

2012-08-01

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Porphyromonas gingivalis Genes Involved in fimA Regulation  

OpenAIRE

Porphyromonas gingivalis is an important component of the complex plaque biofilm that is a direct precursor of periodontal disease. The major fimbriae are required for attachment to oral surfaces and are an important virulence factor. Fimbrillin (FimA) expression in P. gingivalis is inhibited by surface molecule of Streptococcus cristatus, an early colonizer of dental plaque. In this study, differential display PCR was used to identify P. gingivalis genes that are regulated in response to S. ...

Xie, Hua; Kozlova, Natalia; Lamont, Richard J.

2004-01-01

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Evidence for the absence of hyaluronidase activity in Porphyromonas gingivalis.  

OpenAIRE

The aim of the present study was to evaluate the ability of Porphyromonas gingivalis to degrade hyaluronic acid. No hyaluronidase activity was detected using a turbidimetric method, whereas a standard plate assay showed a positive reaction for P. gingivalis. We postulated that the high proteolytic activity of P. gingivalis may account for this observation. A modified plate assay was designed to avoid false-positive reactions caused by proteolytic bacteria. The new assay, based on the formatio...

Grenier, D.; Michaud, J.

1993-01-01

25

Porphyromonas gingivalis Cysteine Proteinase Inhibition by ?-Casein Peptides ?  

OpenAIRE

Porphyromonas gingivalis is a major pathogen associated with chronic periodontitis, an inflammatory disease of the supporting tissues of the teeth. The Arg-specific (RgpA/B) and Lys-specific (Kgp) cysteine proteinases of P. gingivalis are major virulence factors for the bacterium. In this study ?-casein(109-137) was identified in a chymosin digest of casein as an inhibiting peptide of the P. gingivalis proteinases. The peptide was synthesized and shown to inhibit proteolytic activity associa...

Toh, Elena C. Y.; Dashper, Stuart G.; Huq, N. Laila; Attard, Troy J.; O Brien-simpson, Neil M.; Chen, Yu-yen; Cross, Keith J.; Stanton, David P.; Paolini, Rita A.; Reynolds, Eric C.

2010-01-01

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Arginine deiminase inhibits Porphyromonas gingivalis surface attachment.  

Science.gov (United States)

The oral cavity is host to a complex microbial community whose maintenance depends on an array of cell-to-cell interactions and communication networks, with little known regarding the nature of the signals or mechanisms by which they are sensed and transmitted. Determining the signals that control attachment, biofilm development and outgrowth of oral pathogens is fundamental to understanding pathogenic biofilm development. We have previously identified a secreted arginine deiminase (ADI) produced by Streptococcus intermedius that inhibited biofilm development of the commensal pathogen Porphyromonas gingivalis through downregulation of genes encoding the major (fimA) and minor (mfa1) fimbriae, both of which are required for proper biofilm development. Here we report that this inhibitory effect is dependent on enzymic activity. We have successfully cloned, expressed and defined the conditions to ensure that ADI from S. intermedius is enzymically active. Along with the cloning of the wild-type allele, we have created a catalytic mutant (ADIC399S), in which the resulting protein is not able to catalyse the hydrolysis of l-arginine to l-citrulline. P. gingivalis is insensitive to the ADIC399S catalytic mutant, demonstrating that enzymic activity is required for the effects of ADI on biofilm formation. Biofilm formation is absent under l-arginine-deplete conditions, and can be recovered by the addition of the amino acid. Taken together, the results indicate that arginine is an important signal that directs biofilm formation by this anaerobe. Based on our findings, we postulate that ADI functions to reduce arginine levels and, by a yet to be identified mechanism, signals P. gingivalis to alter biofilm development. ADI release from the streptococcal cell and its cross-genera effects are important findings in understanding the nature of inter-bacterial signalling and biofilm-mediated diseases of the oral cavity. PMID:23242802

Cugini, Carla; Stephens, Danielle N; Nguyen, Daniel; Kantarci, Alpdogan; Davey, Mary E

2013-02-01

27

Purificación y caracterización de lipopolisacáridos de Eikenella corrodens 23834 y Porphyromonas gingivalis W83  

OpenAIRE

Título corto: Metodología para el aislamiento e identificación  de LPS de periodonto-patógenosTítulo en inglés: Purification and characterization of lipopolysaccharide from Eikenella corrodens 23834 and Porphyromonas gingivalis W83Resumen: 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 enfe...

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

2014-01-01

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Interleukin-1alpha stimulation in monocytes by periodontal bacteria: antagonistic effects of Porphyromonas gingivalis.  

Science.gov (United States)

Periodontal pathogenic bacteria are associated with elevated levels of interleukin-1alpha (IL-1alpha) but it is unclear if all species can induce cytokine production equally. Porphyromonas gingivalis may be able antagonize IL-1alpha induced by other species through the activity of its proteases or lipopolysaccharide (LPS). Monomac-6 cells and primary human monocytes were treated with culture supernatants from Porphyromonas gingivalis, Fusobacterium nucleatum, Campylobacter rectus, Actinobacillus actinomycetemcomitans, Prevotella intermedius, Veillonella atypical and Prevotella nigrescens. IL-1alpha protein levels were measured after 6 h of incubation. In addition, monocytes were co-stimulated with supernatants from P. gingivalis and other bacteria. The role of P. gingivalis proteases was tested using Arg-X and Lys-X mutant strains. The role of LPS was investigated using purified P. gingivalis LPS and polymixin depletion. All species tested induced significant IL-1alpha production, but P. gingivalis was the weakest. Co-stimulation of monocytes with P. gingivalis antagonized the ability of other bacterial species to induce IL-1alpha production. This effect was at its greatest with C. rectus (resulting in a 70% reduction). Gingipain mutant strains and chemical inhibition of protease activity did not reduce antagonistic activity. However, 100 ng/ml of P. gingivalis LPS can reproduce the antagonistic activity of P. gingivalis culture supernatants. Periodontitis-associated bacterial species stimulate IL-1alpha production by monocytes. P. gingivalis can antagonize this effect, and its LPS appears to be the crucial component. This study highlights the importance of mixed infections in the pathogenesis of periodontal disease because reduction of pro-inflammatory cytokine levels may impair the ability of the host to tackle infection. PMID:17241171

Bostanci, N; Allaker, R; Johansson, U; Rangarajan, M; Curtis, M A; Hughes, F J; McKay, I J

2007-02-01

29

Porphyromonas gingivalis antagonises Campylobacter rectus induced cytokine production by human monocytes.  

Science.gov (United States)

Porphyromonas gingivalis and Campylobacter rectus are two major bacterial species implicated in the pathogenesis of periodontitis. P. gingivalis can antagonise the inflammatory response to other periodontal pathogens, a property commonly attributed to its lipopolysaccharide (LPS). The aim of this study was to investigate the capacity of P. gingivalis to antagonise C. rectus induced cytokine stimulation from human monocytes, and to investigate the involvement of its LPS. Primary human monocytes and Monomac-6 cells were challenged with culture supernatants from P. gingivalis and C. rectus, and levels of IL-1beta, IL-6 and IL-8 produced were measured by ELISA after 6h incubation. Purified P. gingivalis LPS was also added alone or in combination with C. rectus culture supernatant. Both species significantly stimulated the production of all three cytokines from the two cell lines, but P. gingivalis was considerably weaker inducer. Co-stimulation of the cells with P. gingivalis and C. rectus suppressed the cytokine-stimulatory capacity of the latter. P. gingivalis LPS alone was sufficient to antagonise IL-6 and IL-8, but not IL-1beta stimulation by C. rectus. In conclusion, mixed infections may impair host immune responses by reducing pro-inflammatory cytokine levels, which may be of relevance to the pathogenesis of periodontitis. PMID:17709256

Bostanci, N; Allaker, R P; Belibasakis, G N; Rangarajan, M; Curtis, M A; Hughes, F J; McKay, I J

2007-08-01

30

Porphyromonas gingivalis culture supernatants differentially regulate interleukin-1beta and interleukin-18 in human monocytic cells.  

Science.gov (United States)

Porphyromonas gingivalis is a major bacterial species implicated in chornic periodontitis, a disease characterized by inflammatory destruction of the tooth supporting tissues. Its main virulence factors are lipopolysaccharide (LPS) and gingipains, a group of cysteine proteinases. Interleukin (IL)-18 is a potent pro-inflammatory cytokine with structural similarities to IL-1beta. This study aimed to investigate if P .gingivalis regulates IL-1beta and IL-18 in monocytic cells. Monomac-6 cells were challenged with P. gingivalis culture supernatants. Quantitative real-time PCR and ELISA were used to investigate IL-1beta and IL-18 mRNA expression and protein secretion, respectively. P. gingivalis enhanced IL-1beta and IL-18 mRNA expression, the former being induced earlier, but transiently. IL-18 up-regulation was not affected by P. gingivalis heat-inactivation or chemical inhibition of its gingipains, whereas both treatments resulted in 50% reduction of IL-1beta expression. Purified P. gingivalis LPS enhanced both IL-1beta and IL-18 expression. However, only IL-1beta, but not IL-18, secretion was detected, and was up-regulated by P. gingivalis. In conclusion, although IL-1beta and IL-18 belong to the same cytokine family, their gene expression and secretion are differentially regulated in human monocytic cells in response to P. gingivalis. Therefore, cytokines of the IL-1 family may participate via different pathways in the complex pathogenesis of periodontitis. PMID:19091595

Hamedi, M; Belibasakis, G N; Cruchley, A T; Rangarajan, M; Curtis, M A; Bostanci, N

2009-02-01

31

Designing a peptide-based vaccine against Porphyromonas gingivalis.  

Science.gov (United States)

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

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

2013-01-01

32

Environmental regulation of fimbrial gene expression in Porphyromonas gingivalis.  

OpenAIRE

Porphyromonas gingivalis fimbriae are an important virulence factor involved in attachment and invasion. Fimbrillin, encoded by the fimA gene, is the major subunit protein of the fimbriae. To elucidate the influence of environmental signals on the expression of the fimA gene, a strain of P. gingivalis (designated PLE) containing a chromosomal transcriptional fusion between a promoterless lacZ gene and the fimA promoter region was constructed. Promoter activity was assessed by measurement of b...

Xie, H.; Cai, S.; Lamont, R. J.

1997-01-01

33

Promoter Architecture of the Porphyromonas gingivalis Fimbrillin Gene  

OpenAIRE

Porphyromonas gingivalis fimbriae can mediate adherence to many of the available substrates in the oral cavity. Expression of P. gingivalis fimbriae is regulated at the transcriptional level by environmental signals, such as temperature and hemin concentration. The arrangement of the upstream promoter and regulatory sequences required for transcription and control of the fimbrial structural gene (fimA) was investigated. Primer extension analysis demonstrated that the transcriptional start sit...

Xie, Hua; Lamont, Richard J.

1999-01-01

34

Regulation of the Porphyromonas gingivalis fimA (Fimbrillin) Gene  

OpenAIRE

In common with many bacterial virulence genes, the fimbrillin (fimA) gene of Porphyromonas gingivalis is modulated in response to environmental fluctuation. The trans-acting components that comprise the regulatory system for transcriptional activity of the fimA gene in P. gingivalis were investigated. Three major proteins were found to bind to the upstream region of the fimA promoter. One of these proteins was fimbrillin itself, and the other two were a major arginine protease (Rgp) and lysin...

Xie, Hua; Chung, Whasun O.; Park, Yoonsuk; Lamont, Richard J.

2000-01-01

35

Tobacco-induced alterations to Porphyromonas gingivalis-host interactions  

OpenAIRE

Smokers are more susceptible than non-smokers to persistent infection by Porphyromonas gingivalis, a causative agent of periodontitis. Patients who smoke exhibit increased susceptibility to periodontitis and are more likely to display severe disease and be refractory to treatment. Paradoxically, smokers demonstrate reduced clinical inflammation. We show that P. gingivalis cells exposed to cigarette smoke extract (CSE) induce a lower pro-inflammatory response (TNF-?, IL-6, IL12 p40) from mono...

Bagaitkar, Juhi; Williams, Lisa R.; Renaud, Diane E.; Bemakanakere, Manjunatha R.; Scott, David A.; Demuth, Donald R.

2009-01-01

36

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

Directory of Open Access Journals (Sweden)

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

Le Xuan

2009-01-01

37

Antibody-Targeted Lethal Photosensitization of Porphyromonas gingivalis  

OpenAIRE

We have previously demonstrated that Porphyromonas gingivalis is susceptible to killing by toluidine blue O (TBO) when irradiated with light from a helium-neon (HeNe) laser. The aim of this study was to determine whether a TBO-antibody conjugate (Ab-TBO) could be used to specifically target P. gingivalis to lethal photosensitization in the presence of Streptococcus sanguis or human gingival fibroblasts (HGFs). When a mixture of P. gingivalis and S. sanguis was exposed to 4 ?g of TBO/ml and i...

Bhatti, M.; Macrobert, A.; Henderson, B.; Shepherd, P.; Cridland, J.; Wilson, M.

2000-01-01

38

Schisandra chinensis ?-iso-cubebenol induces heme oxygenase-1 expression through PI3K/Akt and Nrf2 signaling and has anti-inflammatory activity in Porphyromonas gingivalis lipopolysaccharide-stimulated macrophages.  

Science.gov (United States)

Heme oxygenase-1 (HO-1) is a potent anti-inflammatory molecule that regulates pro-inflammatory mediators. Several studies have indicated that HO-1 expression is induced by a variety of stimuli such as lipopolysaccharide (LPS), cytokines, oxidative stress, and antioxidant phytochemicals. In this study, we assessed the anti-inflammatory effects of a novel ?-iso-cubebenol isolated from dried fruits of Schisandra chinensis in human macrophage THP-1 cells and investigated the involvement of HO-1 signaling. We first observed that ?-iso-cubebenol induced HO-1 mRNA and protein expression in a dose- and time-dependent manner via activation of erythroid-specific nuclear factor-regulated factor 2 (Nrf2). We also found that ?-iso-cubebenol induced phosphorylation of phosphoinositide 3-kinase (PI3K)/Akt and extracellular-regulated kinase (ERK) in a time-dependent manner. Furthermore, treatment of THP-1 cells with inhibitors and siRNA specific for PI3K/Akt and ERK decreased the expression of HO-1. These results suggested that ?-iso-cubebenol induced HO-1 expression through the activation of PI3K/Akt, ERK, and Nrf2 signaling. Next, ?-iso-cubebenol strongly inhibited Porphyromonas gingivalis LPS-stimulated pro-inflammatory cytokines (tumor necrosis factor (TNF)-?, interleukin (IL)-1?, IL-6, and IL-12). Moreover, we observed that ?-iso-cubebenol treatment inhibited nuclear levels and activity of NF-?B in a dose-dependent manner. Additionally, treatment with tin-protoporphyrin (SnPP), a selective inhibitor of HO-1, reversed the ?-iso-cubebenol-mediated inhibition of P. gingivalis LPS-induced pro-inflammatory cytokines. Hence, ?-iso-cubebenol might induce anti-inflammatory effects on P. gingivalis LPS-stimulated human THP-1 macrophages by mediating the activation of PI3k/Akt and ERK that leads to over-expression of HO-1 and Nrf-2. These findings suggest that ?-iso-cubebenol may be considered as a novel therapeutic agent to ameliorate periodontitis. PMID:21840424

Park, Sun Young; Park, Da Jung; Kim, Young Hun; Kim, YoungHee; Choi, Young-Whan; Lee, Sang-Joon

2011-11-01

39

Doxycycline inhibits TREM-1 induction by Porphyromonas gingivalis.  

Science.gov (United States)

The triggering receptor expressed on myeloid cells 1 (TREM-1) is a cell surface receptor of the immunoglobulin superfamily, with the capacity to amplify pro-inflammatory cytokine production. Porphyromonas gingivalis is a Gram-negative anaerobic species highly implicated in inflammatory periodontal disease, with potential involvement in systemic inflammation. Porphyromonas gingivalis positively regulates TREM-1 expression and production in monocytic cells. Subantimicrobial doses of doxycycline (SDD) are used as an adjunct treatment in periodontal therapy, because of their anti-inflammatory properties. The aim of this study was to investigate the effect of SDD on P. gingivalis-induced TREM-1 expression and secretion by the myelomonocytic cell line MonoMac-6. After 24 h of challenge, P. gingivalis enhanced TREM-1 gene expression by the cells, with a concomitant increase in soluble TREM-1 release. Nevertheless, SDD concentrations between 2 and 10 ?g mL(-1) abolished TREM-1 expression and release, already after 4 h of administration. Moreover, SDD reduced P. gingivalis-induced interleukin-8 secretion, confirming its anti-inflammatory effects. In conclusion, SDD inhibits bacterially induced TREM-1, and this effect may partly account for its generalized anti-inflammatory properties. This could partly explain the clinical efficacy of SDD as an adjunctive treatment for periodontal disease, but may also indicate that SDD could serve as a suitable modulator of systemic inflammatory responses. PMID:22540741

Bostanci, Nagihan; Belibasakis, Georgios N

2012-10-01

40

Identification of interspecies interactions affecting Porphyromonas gingivalis virulence phenotypes  

Directory of Open Access Journals (Sweden)

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

41

Can Porphyromonas gingivalis be a novel aetiology for recurrent miscarriage?  

Science.gov (United States)

Objective To study the association between Porphyromonas gingivalis (P. gingivalis) infection and recurrent miscarriage. Methods This case control study included women with early pregnancy failure admitted for surgical evacuation of retained products of conception. Cases (group 1) included 50 women with unexplained recurrent early miscarriage whereas the control group (group 2) consisted of 50 women with no such history. The evacuated products of conception, subgingival plaques, cervicovaginal secretions and saliva of all participants were examined to detect P. gingivalis deoxyribonucleic acid (DNA) using a polymerase chain reaction. Results The prevalence of P. gingivalis DNA in the chorionic villous tissue samples of group 1 was significantly higher than in group 2 (8 [16%] vs. 1 [2%], respectively; p = 0.036, odds ratio [OR]: 9.3, 95% confidence interval [CI]: 1.1-76.9). The prevalence of P. gingivalis DNA was significantly higher in cervicovaginal secretions of group 1 than in group 2 (9 [18%] vs. 1 [2%], respectively; p = 0.02, OR: 10.8, 95% CI: 1.3-88.5). On the contrary, P. gingivalis DNA could not be detected in subgingival plaques and saliva samples of either group. Conclusion The current study found an association between P. gingivalis infection of the female genital tract and the occurrence of recurrent miscarriage. PMID:25328050

Ibrahim, Moustafa I; Abdelhafeez, Mohamed A; Ellaithy, Mohamed I; Salama, Ahmed H; Amin, Adel S; Eldakrory, Hesham; Elhadad, Nagwa I

2015-04-01

42

Porphyromonas gingivalis and the autophagic pathway: an innate immune interaction?  

Science.gov (United States)

Autophagy is a mechanism used to maintain several intracellular functions essential to eukaryotic cells. Recently, a role for autophagy in innate and adaptive immunity has also been established including the elimination of invading bacteria. Although some intracellular pathogens are killed by autophagy, several others subvert autophagy to the pathogen's benefit for survival and replication. Porphyromonas gingivalis, an important periodontal pathogen, has been shown to stimulate autophagy in endothelial cells and to use the autophagic pathway to its advantage. In human coronary artery endothelial cells (HCAEC), P. gingivalis localizes within autophagosomes. After intracellular uptake, P. gingivalis transits from early autophagosomes to late autophagosomes and prevents the formation of autolysosomes, either by delaying the autophagosome-lysosome fusion or by redirecting the normal autophagic trafficking. In addition, P. gingivalis was also found to stimulate autophagy in human aortic endothelial cells (HAEC) since co-localization of LC3-II, an autophagosome marker, with P. gingivalis was observed. The trafficking of P. gingivalis into the autophagic pathway appears to be dependent upon the host cell type. Survival of P. gingivalis through the subversion of the host autophagic pathway can be considered a bacterial strategy to evade the innate immune system and persist in the host. PMID:17981536

Rodrigues, Paulo Henrique; Bélanger, Myriam; Dunn, William; Progulske-Fox, Ann

2008-01-01

43

Isolation and characterization of a minor fimbria from Porphyromonas gingivalis.  

OpenAIRE

We have discovered two distinctly different fimbriae expressed by the same Porphyromonas gingivalis strain. The construction of a fimA mutant of P. gingivalis ATCC 33277 has previously been reported by N. Hamada et al. (Infect. Immun. 62:1696-1704, 1994). Expression of fimbriae on the surface of the fimA mutant and the wild-type strain, ATCC 33277, were investigated by electron microscopy. The wild-type strain produced long fimbrial structures extending from the cell surface, whereas those st...

Hamada, N.; Sojar, H. T.; Cho, M. I.; Genco, R. J.

1996-01-01

44

Osteomyelitis of the Ulna Caused by Porphyromonas gingivalis  

OpenAIRE

A 41-year-old man was provided with a jacket crown after a root end resection of a molar. Four months later, cortical destruction of the ulnar diaphysis with swelling and pain appeared in his forearm. No microorganism could be grown from an intraoperative tissue specimen, but bacterial 16S rRNA genes were detected by broad-range PCR, revealing Porphyromonas gingivalis as the causative agent of osteomyelitis.

Welkerling, Heike; Geißdo?rfer, Walter; Aigner, Thomas; Forst, Raimund

2006-01-01

45

Benzamidine derivatives inhibit the virulence of Porphyromonas gingivalis  

OpenAIRE

We have previously shown that benzamidine-type compoundscan inhibit the activity of arginine-specific cysteine proteinases (gingipainsHRgpA and RgpB); well-known virulence factors of Porphyromonas gingivalis. They also hinder in vitro growth of this important period on to pathogenic bacterium. Apparently growth arrest is not associated with their ability to inhibit these proteases, as pentamidine, which is a 20-fold less efficient inhibitor of gingipainthan 2,6-bis-(4-amidinobenzyl)-cyclohexa...

Fro?hlich, Esther; Kantyka, Tomasz; Plaza, Karolina; Schmidt, Karl-hermann; Pfister, Wolfgang; Potempa, Jan; Eick, Sigrun

2012-01-01

46

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

OpenAIRE

To characterize the roles of Porphyromonas gingivalis and its components in disease processes, we investigated the cytokine profiles induced by live P. gingivalis, its lipopolysaccharide (LPS), and its major fimbrial protein, fimbrillin (FimA). A cytokine antibody array revealed that human monocyte-derived macrophages were induced to produce chemokines (e.g., monocyte chemoattractant protein 1, macrophage inflammatory protein 1? [MIP-1?], and MIP-3?) as early as 1 h after exposure to P. gi...

Zhou, Qingde; Amar, Salomon

2006-01-01

47

Porphyromonas gingivalis Infection Reduces Regulatory T Cells in Infected Atherosclerosis Patients  

OpenAIRE

Increasing evidence has shown periodontal pathogen Porphyromonas gingivalis (P.gingivalis) infection contributes to atherosclerosis (AS) progression. P.gingivalis fimbriae act as an important virulence factor in AS. Regulatory T cells (Tregs) may play a crucial role in autoimmune response during this process. However, whether P.gingivalis infection is associated with Tregs dysregulation during AS is still unknown and the prevalence of different P.gingivalis FimA genotypes during this process ...

Yang, Jie; Wu, Juan; Liu, Yu; Huang, Jin; Lu, Zhipin; Xie, Liping; Sun, Weibin; Ji, Yong

2014-01-01

48

Oral Immunization with Recombinant Streptococcus gordonii Expressing Porphyromonas gingivalis FimA Domains  

OpenAIRE

Porphyromonas gingivalis, a gram-negative anaerobe, is implicated in the etiology of adult periodontitis. P. gingivalis fimbriae are one of several critical surface virulence factors involved in both bacterial adherence and inflammation. P. gingivalis fimbrillin (FimA), the major subunit protein of fimbriae, is considered an important antigen for vaccine development against P. gingivalis-associated periodontitis. We have previously shown that biologically active domains of P. gingivalis fimbr...

Sharma, Ashu; Honma, Kiyonobu; Evans, Richard T.; Hruby, Dennis E.; Genco, Robert J.

2001-01-01

49

Porphyromonas gingivalis promotes Th17 inducing pathways in chronic periodontitis.  

Science.gov (United States)

In periodontitis, a common chronic inflammatory condition, gram-negative-rich bacterial biofilms trigger, in susceptible individuals, perpetuating inflammation that results in extensive tissue damage of tooth supporting structures. To delineate immune cell-dependent mechanisms whereby bacterial challenge drives persistent destructive inflammation in periodontitis and other inflammatory diseases, we studied involved tissues ex vivo and investigated host cell responses to the periodontal pathogen Porphyromonas gingivalis, in vitro. Diseased lesions were populated by abundant Th17 cells, linked to infection, chronic inflammation/autoimmunity and tissue pathology. In vitro, P. gingivalis, particularly the more virulent strain W83, stimulated myeloid antigen presenting cells (APC) to drive Th17 polarization. Supernatants from myeloid APC exposed to P. gingivalis were capable of enhancing Th17 but not Th1 polarization. P. gingivalis favored the generation of Th17 responses by stimulating the production of Th17 related cytokines IL-1?, IL-6 and IL-23, but not Th1 related IL-12. By inducing NF?B activation, P. gingivalis promoted IL-1?, IL-6 and IL-12p40 production, but not IRF3 phosphorylation, connected to generation of the IL-12p35 chain, ultimately restricting formation of the intact IL-12 molecule. Promotion of Th17 lineage responses was also aided by P. gingivalis proteases, which appeared to differentially degrade pivotal cytokines. In this regard, IL-12 was largely degraded by P. gingivalis, whereas IL-1? was more resistant to proteolysis. Our data unveil multiple pathways by which P. gingivalis may orchestrate chronic inflammation, providing insights into interventional strategies. PMID:22560973

Moutsopoulos, Niki M; Kling, Heather M; Angelov, Nikola; Jin, Wenwen; Palmer, Robert J; Nares, Salvador; Osorio, Manuel; Wahl, Sharon M

2012-12-01

50

Adherence of Porphyromonas (Bacteroides) gingivalis to Streptococcus sanguis in vitro.  

Science.gov (United States)

Intergeneric bacterial adherence is responsible for the complexity of the microbiota in human dental plaque and is believed to enable some extraneous bacteria to initially colonize the human oral cavity. Some current evidence indicates that Streptococcus sanguis, an early colonizer of teeth, enhances subsequent colonization by Porphyromonas (Bacteroides) gingivalis, a bacterium associated with advanced adult periodontitis. In this study, selected strains of P. gingivalis and S. sanguis were tested for their adherence activities in vitro. A differential filtration assay was devised in which one member of the test pair was radiolabeled. Heterogeneous aggregates that formed in mixed suspensions were collected on polycarbonate filters (8-microns pore size) and were washed free of individual bacteria and small homologous clumps. P. gingivalis 381, W50, JKG7, and 33277 adhered to S. sanguis G9B, M5, Challis 6, and 38. P. gingivalis A7A1-28 did not adhere well to S. sanguis under these conditions. More precise measurements of intergeneric adherence were obtained with an alternative assay with radiolabeled P. gingivalis and an artificial dental plaque composed of S. sanguis coupled to cyanogen bromide-activated agarose beads. CNBr-agarose was selected as the supporting matrix for the plaque because it was uniformly and permanently coated with S. sanguis and because P. gingivalis had negligible adherence activity for streptococcus-free beads. P. gingivalis W50 grown to the early stationary phase adhered to S. sanguis-coated beads in higher numbers than either midlogarithmic- or late-stationary-phase cells. Intergeneric adherence was not inhibited or reversed by the presence of lactose or other monosaccharides or disaccharides. Pretreatment of either bacterium with trypsin or proteinase K reduced subsequent adherence by 86 to 100%. Neuraminidase treatment of P. gingivalis caused 98% reduction of adherence, whereas similar treatment of S. sanguis caused only a 2% loss. Preincubation of P. gingivalis at 60 degrees C for 30 min decreased subsequent adherence to S. sanguis-coated beads by 94%. Adherence was reduced by 96% when bacteria were assayed while suspended in human whole saliva or when pretreated with saliva and subsequently assayed in buffer. The concentration of whole human saliva required to inhibit 50% adherence in this assay was 23 micrograms per ml (1:200 dilution). Suspension of the bacteria in normal rabbit serum resulted in 94% inhibition of adherence. These data indicate that saliva and serum may be important host defense factors for controlling Porphyromonas-Streptococcus adherence. PMID:1987021

Stinson, M W; Safulko, K; Levine, M J

1991-01-01

51

Lineage variability in surface components expression within Porphyromonas gingivalis.  

Science.gov (United States)

The periodontopathogen Porphyromonas gingivalis is represented by a spectrum of phenotypes ranging from commensals to pathogenic lineages. Capsule and fimbriae are considered key virulence factors in this specie, involved in colonization and host defenses evasion. Since these virulence traits may not be expressed by certain strains, we aimed to test the hypothesis that certain clusters or genotypes of P. gingivalis correlate with the production of capsule and fimbriae. Sixteen P. gingivalis isolates were evaluated. Capsule (K) was detected by optical microscopy of negatively stained cells. The presence of fimbriae (F) was determined by TEM. Genotypes were determined by NotI macrorestriction fragments analysis through Pulsed-Field Gel Electrophoresis (PFGE) and Multi-locus sequence typing (MLST) based on seven house-keeping genes. The phenotypes included F(+)K(+) (n = 4), F(-)K(+) (n = 5), F(+)K(-) (n = 5) and F(-)K(-) (n = 2). The analysis of whole genome macrorestriction fragments revealed 14 different clusters. MLST data also revealed extensive genetic diversity; however, PFGE and MLST profiles showed evident differences. There was no association between P. gingivalis clusters and encapsulated and/or fimbriated phenotypes. Genotyping methods were not able to discriminate isolates according to the production of virulence factors such as capsule and major fimbriae, indicating that recombination played a key role in the expression of capsule and fimbriae in P. gingivalis. PMID:25448131

Teixeira, Silvia Regina Loureiro; D'Epiro, Talyta Thereza Soares; Pinheiro, Ericka Tavares; Simionato, Maria Regina L; Taniwaki, Noemi Nosomi; Kisielius, Jonas José; Mayer, Marcia Pinto Alves

2014-12-01

52

Porphyromonas gingivalis Cysteine Proteinase Inhibition by ?-Casein Peptides ?  

Science.gov (United States)

Porphyromonas gingivalis is a major pathogen associated with chronic periodontitis, an inflammatory disease of the supporting tissues of the teeth. The Arg-specific (RgpA/B) and Lys-specific (Kgp) cysteine proteinases of P. gingivalis are major virulence factors for the bacterium. In this study ?-casein(109-137) was identified in a chymosin digest of casein as an inhibiting peptide of the P. gingivalis proteinases. The peptide was synthesized and shown to inhibit proteolytic activity associated with P. gingivalis whole cells, purified RgpA-Kgp proteinase-adhesin complexes, and purified RgpB proteinase. The peptide ?-casein(109-137) exhibited synergism with Zn(II) against both Arg- and Lys-specific proteinases. The active region for inhibition was identified as ?-casein(117-137) using synthetic peptides. Kinetic studies revealed that ?-casein(109-137) inhibits in an uncompetitive manner. A molecular model based on the uncompetitive action and its synergistic ability with Zn(II) was developed to explain the mechanism of inhibition. Preincubation of P. gingivalis with ?-casein(109-137) significantly reduced lesion development in a murine model of infection. PMID:21173178

Toh, Elena C. Y.; Dashper, Stuart G.; Huq, N. Laila; Attard, Troy J.; O'Brien-Simpson, Neil M.; Chen, Yu-Yen; Cross, Keith J.; Stanton, David P.; Paolini, Rita A.; Reynolds, Eric C.

2011-01-01

53

Porphyromonas gingivalis cysteine proteinase inhibition by kappa-casein peptides.  

Science.gov (United States)

Porphyromonas gingivalis is a major pathogen associated with chronic periodontitis, an inflammatory disease of the supporting tissues of the teeth. The Arg-specific (RgpA/B) and Lys-specific (Kgp) cysteine proteinases of P. gingivalis are major virulence factors for the bacterium. In this study ?-casein(109-137) was identified in a chymosin digest of casein as an inhibiting peptide of the P. gingivalis proteinases. The peptide was synthesized and shown to inhibit proteolytic activity associated with P. gingivalis whole cells, purified RgpA-Kgp proteinase-adhesin complexes, and purified RgpB proteinase. The peptide ?-casein(109-137) exhibited synergism with Zn(II) against both Arg- and Lys-specific proteinases. The active region for inhibition was identified as ?-casein(117-137) using synthetic peptides. Kinetic studies revealed that ?-casein(109-137) inhibits in an uncompetitive manner. A molecular model based on the uncompetitive action and its synergistic ability with Zn(II) was developed to explain the mechanism of inhibition. Preincubation of P. gingivalis with ?-casein(109-137) significantly reduced lesion development in a murine model of infection. PMID:21173178

Toh, Elena C Y; Dashper, Stuart G; Huq, N Laila; Attard, Troy J; O'Brien-Simpson, Neil M; Chen, Yu-Yen; Cross, Keith J; Stanton, David P; Paolini, Rita A; Reynolds, Eric C

2011-03-01

54

Sialidase and Sialoglycoproteases Can Modulate Virulence in Porphyromonas gingivalis ? †  

Science.gov (United States)

The Porphyromonas gingivalis recombinant VimA can interact with the gingipains and several other proteins, including a sialidase. Sialylation can be involved in protein maturation; however, its role in virulence regulation in P. gingivalis is unknown. The three sialidase-related proteins in P. gingivalis showed the characteristic sialidase Asp signature motif (SXDXGXTW) and other unique domains. To evaluate the roles of the associated genes, randomly chosen P. gingivalis isogenic mutants created by allelic exchange and designated FLL401 (PG0778::ermF), FLL402 (PG1724::ermF), and FLL403 (PG0352::ermF-ermAM) were characterized. Similar to the wild-type strain, FLL402 and FLL403 displayed a black-pigmented phenotype in contrast to FLL401, which was not black pigmented. Sialidase activity in P. gingivalis FLL401 was reduced by approximately 70% in comparison to those in FLL402 and FLL403, which were reduced by approximately 42% and 5%, respectively. Although there were no changes in the expression of the gingipain genes, their activities were reduced by 60 to 90% in all the isogenic mutants compared to that for the wild type. Immunoreactive bands representing the catalytic domains for RgpA, RgpB, and Kgp were present in FLL402 and FLL403 but were missing in FLL401. While adhesion was decreased, the capacity for invasion of epithelial cells by the isogenic mutants was increased by 11 to 16% over that of the wild-type strain. Isogenic mutants defective in PG0778 and PG0352 were more sensitive to hydrogen peroxide than the wild type. Taken together, these results suggest that the P. gingivalis sialidase activity may be involved in regulating gingipain activity and other virulence factors and may be important in the pathogenesis of this organism. PMID:21502589

Aruni, Wilson; Vanterpool, Elaine; Osbourne, Devon; Roy, Francis; Muthiah, Arun; Dou, Yuetan; Fletcher, Hansel M.

2011-01-01

55

Sialidase and sialoglycoproteases can modulate virulence in Porphyromonas gingivalis.  

Science.gov (United States)

The Porphyromonas gingivalis recombinant VimA can interact with the gingipains and several other proteins, including a sialidase. Sialylation can be involved in protein maturation; however, its role in virulence regulation in P. gingivalis is unknown. The three sialidase-related proteins in P. gingivalis showed the characteristic sialidase Asp signature motif (SXDXGXTW) and other unique domains. To evaluate the roles of the associated genes, randomly chosen P. gingivalis isogenic mutants created by allelic exchange and designated FLL401 (PG0778::ermF), FLL402 (PG1724::ermF), and FLL403 (PG0352::ermF-ermAM) were characterized. Similar to the wild-type strain, FLL402 and FLL403 displayed a black-pigmented phenotype in contrast to FLL401, which was not black pigmented. Sialidase activity in P. gingivalis FLL401 was reduced by approximately 70% in comparison to those in FLL402 and FLL403, which were reduced by approximately 42% and 5%, respectively. Although there were no changes in the expression of the gingipain genes, their activities were reduced by 60 to 90% in all the isogenic mutants compared to that for the wild type. Immunoreactive bands representing the catalytic domains for RgpA, RgpB, and Kgp were present in FLL402 and FLL403 but were missing in FLL401. While adhesion was decreased, the capacity for invasion of epithelial cells by the isogenic mutants was increased by 11 to 16% over that of the wild-type strain. Isogenic mutants defective in PG0778 and PG0352 were more sensitive to hydrogen peroxide than the wild type. Taken together, these results suggest that the P. gingivalis sialidase activity may be involved in regulating gingipain activity and other virulence factors and may be important in the pathogenesis of this organism. PMID:21502589

Aruni, Wilson; Vanterpool, Elaine; Osbourne, Devon; Roy, Francis; Muthiah, Arun; Dou, Yuetan; Fletcher, Hansel M

2011-07-01

56

Intra- and inter-individual comparison of Porphyromonas gingivalis genotypes.  

Science.gov (United States)

Genetic analysis of 31 clinical strains of Porphyromonas gingivalis isolated from nine subjects, 2-6 strains per subject, was performed by Southern hybridization. Chromosomal DNA was extracted by the method of Moncla et al. [1] and digested to completion with restriction endonucleases PstI, ClaI and BglI. The DNA fragments were separated electrophoretically on agarose gels, transferred to nylon membranes and hybridized to the non-radioactively labelled plasmid pKK 3535 which contains the rmB ribosomal RNA operon of the Escherichia coli chromosome. Of the three enzymes, BglI was the most suitable for the genetic analysis of P. gingivalis. With this enzyme, the intra-individual strains were shown to be identical in eight of the nine subjects, whereas inter-individual strains were different. PMID:8390897

Saarela, M; Stucki, A M; von Troil-Lindén, B; Alaluusua, S; Jousimies-Somer, H; Asikainen, S

1993-03-01

57

Role of fimbriae in Porphyromonas gingivalis invasion of gingival epithelial cells.  

OpenAIRE

Porphyromonas gingivalis is a periodontal pathogen capable of invading primary cultures of normal human gingival epithelial cells (NHGEC). Involvement of P. gingivalis fimbriae in the invasion process was examined. Purified P. gingivalis 33277 fimbriae blocked invasion of this organism into NHGEC in a dose-dependent manner. DPG3, a P. gingivalis fimbria-deficient mutant, was impaired in its invasion capability approximately eightfold compared to its parent, strain 381. However, adherence of t...

Weinberg, A.; Belton, C. M.; Park, Y.; Lamont, R. J.

1997-01-01

58

Purificación y caracterización de lipopolisacáridos de Eikenella corrodens 23834 y Porphyromonas gingivalis W83  

Directory of Open Access Journals (Sweden)

Full Text Available Título corto: Metodología para el aislamiento e identificación  de LPS de periodonto-patógenosTítulo en inglés: Purification and characterization of lipopolysaccharide from Eikenella corrodens 23834 and Porphyromonas gingivalis W83Resumen: 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 estudio 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.Palabras clave: endotoxinas, cromatografía, ácido 2-ceto-3-desoxioctulosónico (KDO, test LAL, periodontitis.Abstract: Purification of lipopolysaccharide (LPS or endotoxins and its characterization is an important aspect for studies aimed at clarifying 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 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 compared with commercial LPS.Key words: endotoxin, 2-keto-3-deoxyoctonate (KDO, Chromatography, Limulus test (LAL, periodontitis.

Diego Fernando Gualtero Escobar

2014-06-01

59

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

OpenAIRE

The effects of histatins on coaggregation between Porphyromonas gingivalis 381 and Streptococcus mitis ATCC 9811 were investigated by using a turbidimetric assay. The coaggregation activity was significantly inhibited by histatins 5 and 8 and strongly by lysozyme. Tritium-labeled histatin 8 bound to P. gingivalis cells but not to S. mitis cells.

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

1991-01-01

60

Role of the Amino-Terminal Region of Porphyromonas gingivalis Fimbriae in Adherence to Epithelial Cells  

OpenAIRE

Porphyromonas gingivalis fimbriae elicit many responses in eukaryotic cells, including mitogenicity, cytokine production, epithelial cell invasion, and cellular immune response. Specific domains of the major fimbrial protein (FimA) have been shown to be important in triggering some of these functions. The goal of the present study was to identify the domain(s) of P. gingivalis FimA responsible for specific interaction with human mucosal epithelial cells. Fimbriated P. gingivalis strains have ...

Sojar, Hakimuddin T.; Han, Yiping; Hamada, Nobushiro; Sharma, Ashu; Genco, Robert J.

1999-01-01

61

Role of Arginine Deiminase of Streptococcus cristatus in Porphyromonas gingivalis Colonization?  

OpenAIRE

The ability to attach to a variety of oral surfaces is an important characteristic of Porphyromonas gingivalis. Previous studies have demonstrated that expression and production of FimA, a major subunit protein of the long fimbriae, is required for P. gingivalis colonization. Here we report that a surface protein, arginine deiminase (ArcA) of Streptococcus cristatus, represses FimA production and inhibits biofilm formation of P. gingivalis. This inhibitory function of ArcA is also observed in...

Wu, Jie; Xie, Hua

2010-01-01

62

Resistance of a Tn4351-generated polysaccharide mutant of Porphyromonas gingivalis to polymorphonuclear leukocyte killing.  

OpenAIRE

In this study, we describe the development of an efficient transpositional mutagenesis system for Porphyromonas gingivalis using the Bacteroides fragilis transposon Tn4351. Using this system, we have isolated and characterized a Tn4351-generated mutant of P. gingivalis A7436, designated MSM-1, which exhibits enhanced resistance to polymorphonuclear leukocyte (PMN) phagocytosis and killing. P. gingivalis MSM-1 was initially selected based on its colony morphology; MSM-1 appeared as a mucoid, b...

Genco, C. A.; Schifferle, R. E.; Njoroge, T.; Forng, R. Y.; Cutler, C. W.

1995-01-01

63

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

OpenAIRE

The aim of the study that we describe was to combine an immunomagnetic separation and a PCR followed by dot blot hybridization with a DNA probe for the detection and identification of Porphyromonas gingivalis. Immunomagnetic particles were coated with monoclonal antibody specific for P. gingivalis and were incubated with a suspension containing seven oral bacterial species spiked with various dilutions of P. gingivalis. Beads with their load of bound bacterial were boiled in water, and the ta...

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

1995-01-01

64

Inactivation of the Porphyromonas gingivalis fimA gene blocks periodontal damage in gnotobiotic rats.  

OpenAIRE

Fimbrial production by Porphyromonas gingivalis was inactivated by insertion-duplication mutagenesis, using the cloned gene for the P. gingivalis major fimbrial subunit protein, fimA. by several criteria, this insertion mutation rendered P. gingivalis unable to produce fimbrilin or an intact fimbrial structure. A nonfimbriated mutant, DPG3, hemagglutinated sheep erythrocytes normally and was unimpaired in the ability to coaggregate with Streptococcus gordonii G9B. The cell surface hydrophobic...

Malek, R.; Fisher, J. G.; Caleca, A.; Stinson, M.; Oss, C. J.; Lee, J. Y.; Cho, M. I.; Genco, R. J.; Evans, R. T.; Dyer, D. W.

1994-01-01

65

Role of vimA in cell surface biogenesis in Porphyromonas gingivalis  

OpenAIRE

The Porphyromonas gingivalis vimA gene has been previously shown to play a significant role in the biogenesis of gingipains. Further, in P. gingivalis FLL92, a vimA-defective mutant, there was increased auto-aggregation, suggesting alteration in membrane surface proteins. In order to determine the role of the VimA protein in cell surface biogenesis, the surface morphology of P. gingivalis FLL92 was further characterized. Transmission electron microscopy demonstrated abundant fimbrial appendag...

Osbourne, Devon O.; Aruni, Wilson; Roy, Francis; Perry, Christopher; Sandberg, Lawrence; Muthiah, Arun; Fletcher, Hansel M.

2010-01-01

66

VimA – dependent modulation of the secretome in Porphyromonas gingivalis  

OpenAIRE

The VimA protein of Porphyromonas gingivalis is a multifunctional protein involved in cell surface biogenesis. To further determine if its acetyl coenzyme A (acetyl-CoA) transfer and putative sorting functions can affect the secretome, its role in peptidoglycan biogenesis and effects on the extracellular proteins of P. gingivalis FLL92, a vimA-defective mutant, were evaluated. There were structural and compositional differences in the peptidoglycan of P. gingivalis FLL92 compared to the wild-...

Osbourne, D.; Aruni, A. Wilson; Dou, Y.; Perry, C.; Boskovic, D. S.; Roy, F.; Fletcher, H. M.

2012-01-01

67

Comparative transcriptomic analysis of Porphyromonas gingivalis biofilm and planktonic cells  

OpenAIRE

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 < 0.01) of the P. gingivalis genome was differentially expresse...

Patricia, Lissel J.; Slakeski Nada; Dashper Stuart G; Boyce John D; Seers Christine A; Lo Alvin W; Reynolds Eric C

2009-01-01

68

Porphyromonas gingivalis Induces Receptor Activator of NF-?B Ligand Expression in Osteoblasts through the Activator Protein 1 Pathway  

Science.gov (United States)

Porphyromonas gingivalis, an important periodontal pathogen, is closely associated with inflammatory alveolar bone resorption, and several components of the organism such as lipopolysaccharides have been reported to stimulate production of cytokines that promote inflammatory bone destruction. We investigated the effect of infection with viable P. gingivalis on cytokine production by osteoblasts. Reverse transcription-PCR and real-time PCR analyses revealed that infection with P. gingivalis induced receptor activator of nuclear factor ?B (NF-?B) ligand (RANKL) mRNA expression in mouse primary osteoblasts. Production of interleukin-6 was also stimulated; however, osteoprotegerin was not. SB20350 (an inhibitor of p38 mitogen-activated protein kinase), PD98059 (an inhibitor of classic mitogen-activated protein kinase kinase, MEK1/2), wortmannin (an inhibitor of phosphatidylinositol 3 kinase), and carbobenzoxyl-leucinyl-leucinyl-leucinal (an inhibitor of NF-?B) did not prevent the RANKL expression induced by P. gingivalis. Degradation of inhibitor of NF-?B-alpha was not detectable; however, curcumin, an inhibitor of activator protein 1 (AP-1), prevented the RANKL production induced by P. gingivalis infection. Western blot analysis revealed that phosphorylation of c-Jun, a component of AP-1, occurred in the infected cells, and an analysis of c-Fos binding to an oligonucleotide containing an AP-1 consensus site also demonstrated AP-1 activation in infected osteoblasts. Infection with P. gingivalis KDP136, an isogenic deficient mutant of arginine- and lysine-specific cysteine proteinases, did not stimulate RANKL production. These results suggest that P. gingivalis infection induces RANKL expression in osteoblasts through AP-1 signaling pathways and cysteine proteases of the organism are involved in RANKL production. PMID:14977979

Okahashi, Nobuo; Inaba, Hiroaki; Nakagawa, Ichiro; Yamamura, Taihei; Kuboniwa, Masae; Nakayama, Koji; Hamada, Shigeyuki; Amano, Atsuo

2004-01-01

69

Porphyromonas gingivalis induces receptor activator of NF-kappaB ligand expression in osteoblasts through the activator protein 1 pathway.  

Science.gov (United States)

Porphyromonas gingivalis, an important periodontal pathogen, is closely associated with inflammatory alveolar bone resorption, and several components of the organism such as lipopolysaccharides have been reported to stimulate production of cytokines that promote inflammatory bone destruction. We investigated the effect of infection with viable P. gingivalis on cytokine production by osteoblasts. Reverse transcription-PCR and real-time PCR analyses revealed that infection with P. gingivalis induced receptor activator of nuclear factor kappaB (NF-kappaB) ligand (RANKL) mRNA expression in mouse primary osteoblasts. Production of interleukin-6 was also stimulated; however, osteoprotegerin was not. SB20350 (an inhibitor of p38 mitogen-activated protein kinase), PD98059 (an inhibitor of classic mitogen-activated protein kinase kinase, MEK1/2), wortmannin (an inhibitor of phosphatidylinositol 3 kinase), and carbobenzoxyl-leucinyl-leucinyl-leucinal (an inhibitor of NF-kappaB) did not prevent the RANKL expression induced by P. gingivalis. Degradation of inhibitor of NF-kappaB-alpha was not detectable; however, curcumin, an inhibitor of activator protein 1 (AP-1), prevented the RANKL production induced by P. gingivalis infection. Western blot analysis revealed that phosphorylation of c-Jun, a component of AP-1, occurred in the infected cells, and an analysis of c-Fos binding to an oligonucleotide containing an AP-1 consensus site also demonstrated AP-1 activation in infected osteoblasts. Infection with P. gingivalis KDP136, an isogenic deficient mutant of arginine- and lysine-specific cysteine proteinases, did not stimulate RANKL production. These results suggest that P. gingivalis infection induces RANKL expression in osteoblasts through AP-1 signaling pathways and cysteine proteases of the organism are involved in RANKL production. PMID:14977979

Okahashi, Nobuo; Inaba, Hiroaki; Nakagawa, Ichiro; Yamamura, Taihei; Kuboniwa, Masae; Nakayama, Koji; Hamada, Shigeyuki; Amano, Atsuo

2004-03-01

70

Characterization of FimA in Porphyromonas gingivalis genotype IV.  

Science.gov (United States)

It has been reported that a large majority of periodontitis patients carry organisms with either type II or IV-fimA, while type I is the most prevalent fimA genotype among Porphyromonas gingivalis-positive healthy adults. Here we report characterization of recombinant fimbrial protein (rFimA) produced in Escherichia coli from genotype IV-fimA. In SDS-PAGE and immunoblot analysis after partial dissociation, type IV-rFimA showed a ladder-like pattern representing oligomeric/polymeric forms of native fimbrial structure. Unlike anti-type I-native fimbriae which can only recognize conformational epitopes of the respective proteins, both anti-type IV-native fimbriae and anti-type IV-rFimA antibodies recognized conformational as well as linear epitopes in type IV-fimbriae. These results suggest that the type IV-rFimA proteins retain the native fimbrial antigenicity and the antigenicity of type IV-fimbriae is different from that of type I-fimbriae. PMID:22429612

Choi, Young-Suk; Moon, Ji-Hoi; Park, Jae-Hong; Lee, Jin-Yong

2012-08-01

71

Infection with Porphyromonas gingivalis Exacerbates Endothelial Injury in Obese Mice  

Science.gov (United States)

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

72

Functional differences of Porphyromonas gingivalis fimbriae in determining periodontal disease pathogenesis: a literature review  

Directory of Open Access Journals (Sweden)

Full Text Available Porphyromonas gingivalis is implicated in chronic and aggressive periodontitis. This bacterium has numerous virulence factors and one is the fimbriae, which is quite important for bacterial colonization. Fimbriae are appendices that anchor to the bacterial wall and are comprised of the protein fimbriline encoded by the fimA gene. Thus far, six genotypes have been identified, fimA I to V and Ib. Genotypes II and IV are associated with periodontal disease, while genotype I is related to gingival health. Genotype identification of P. gingivalis fimA in periodontitis would be important to confirm the pathogenic genotypes and to establish risk at population level. This review is about the P. gingivalis fimA genotype prevalence worldwide. A systematic search using Pubmed, Hinary, and Science Direct within the following descriptors: Porphyromonas gingivalis, bacterial adhesion, periodontitis, fimbriae, fimA, genotipification was performed to April 2011.

Moreno, Sandra

2013-03-01

73

Functional differences of Porphyromonas gingivalis Fimbriae in determining periodontal disease pathogenesis: a literature review.  

Science.gov (United States)

Porphyromonas gingivalis is implicated in chronic and aggressive periodontitis. This bacterium has numerous virulence factors and one is the Fimbriae, which is quite important for bacterial colonization. Fimbriae are appendices that anchor to the bacterial wall and are comprised of the protein FimBriline encoded by the FimA gene. Thus far, six genotypes have been identified, FimA I to V and Ib. Genotypes II and IV are associated with periodontal disease, while genotype I 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. PMID:24892323

Moreno, Sandra; Contreras, Adolfo

2013-01-01

74

Expression of the tpr Protease Gene of Porphyromonas gingivalis Is Regulated by Peptide Nutrients  

OpenAIRE

The Tpr protease of Porphyromonas gingivalis W83 is a membrane-associated enzyme capable of hydrolyzing chromogenic substrates for trypsin and bacterial collagenases. A previous study by us indicated that Tpr expression was increased under conditions of nutrient limitation. In the present study, we further characterized expression of the tpr gene using a tpr::lacZ reporter gene construct under a range of nutrient conditions. In P. gingivalis, transcription of tpr was initiated 215 bp upstream...

Lu, Biqing; Mcbride, Barry C.

1998-01-01

75

Intercellular Spreading of Porphyromonas gingivalis Infection in Primary Gingival Epithelial Cells  

OpenAIRE

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 organi...

Yilmaz, O?zlem; Verbeke, Philippe; Lamont, Richard J.; Ojcius, David M.

2006-01-01

76

Conjugal Transfer of Chromosomal DNA Contributes to Genetic Variation in the Oral Pathogen Porphyromonas gingivalis?  

OpenAIRE

Porphyromonas gingivalis is a major oral pathogen that contributes to the development of periodontal disease. There is a significant degree of genetic variation among strains of P. gingivalis, and the population structure has been predicted to be panmictic, indicating that horizontal DNA transfer and recombination between strains are likely. The molecular events underlying this genetic exchange are not understood, although a putative type IV secretion system is present in the genome sequence ...

Tribble, Gena D.; Lamont, Gwyneth J.; Progulske-fox, Ann; Lamont, Richard J.

2007-01-01

77

Mice Lacking Inducible Nitric Oxide Synthase Demonstrate Impaired Killing of Porphyromonas gingivalis  

OpenAIRE

Porphyromonas gingivalis is a primary etiological agent of generalized severe periodontitis, and emerging data suggest the importance of reactive oxygen and nitrogen species in periodontal tissue damage, as well as in microbial killing. Since nitric oxide (NO) released from inducible NO synthase (iNOS) has been shown to possess immunomodulatory, cytotoxic, and antibacterial effects in experimental models, we challenged iNOS-deficient (iNOS?/?) mice with P. gingivalis by using a subcutaneo...

Gyurko, Robert; Boustany, Gabriel; Huang, Paul L.; Kantarci, Alpdogan; Dyke, Thomas E.; Genco, Caroline A.; Gibson Iii, Frank C.

2003-01-01

78

Hemoglobinase Activity of the Lysine Gingipain Protease (Kgp) of Porphyromonas gingivalis W83  

OpenAIRE

Porphyromonas gingivalis, an important periodontal disease pathogen, forms black-pigmented colonies on blood agar. Pigmentation is believed to result from accumulation of iron protoporphyrin IX (FePPIX) derived from erythrocytic hemoglobin. The Lys-X (Lys-gingipain) and Arg-X (Arg-gingipain) cysteine proteases of P. gingivalis bind and degrade erythrocytes. We have observed that mutations abolishing activity of the Lys-X-specific cysteine protease, Kgp, resulted in loss of black pigmentation ...

Lewis, Janina P.; Dawson, Janet A.; Hannis, James C.; Muddiman, David; Macrina, Francis L.

1999-01-01

79

Sodium Ion-Driven Serine/Threonine Transport in Porphyromonas gingivalis  

OpenAIRE

Porphyromonas gingivalis is an asaccharolytic, gram-negative bacterium that relies on the fermentation of amino acids for metabolic energy. When grown in continuous culture in complex medium containing 4 mM (each) free serine, threonine, and arginine, P. gingivalis assimilated mainly glutamate/glutamine, serine, threonine, aspartate/asparagine, and leucine in free and/or peptide form. Serine and threonine were assimilated in approximately equal amounts in free and peptide form. We characteriz...

Dashper, S. G.; Brownfield, L.; Slakeski, N.; Zilm, P. S.; Rogers, A. H.; Reynolds, E. C.

2001-01-01

80

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

OpenAIRE

Objectives. To examine the interaction between Porphyromonas gingivalis and 3 different orthodontic brackets in vitro, focusing on the effect of an early salivary pellicle and other bacteria on the formation of biofilms. Material and Methods. Mono- and multi-species P. gingivalis biofilms were allowed to form in vitro, on 3 different bracket types (stainless steel, ceramic and plastic) with and without an early salivary pellicle. The brackets were anaerobically incubated for 3 days in Brain H...

William Papaioannou; Athanasios Panagopoulos; Haroula Koletsi-Kounari; Efterpi Kontou; Margarita Makou

2012-01-01

81

Immunization against Porphyromonas gingivalis inhibits progression of experimental periodontitis in nonhuman primates.  

OpenAIRE

Periodontitis is a common infectious disease in which the attachment tissues of the teeth and their alveolar bone housing are destroyed, resulting in tooth loss. The gram-negative anaerobic microorganism Porphyromonas gingivalis has been closely linked to severe forms of the disease. We show for the first time that immunization of the primate Macaca fascicularis with killed P. gingivalis in Syntex Adjuvant Formulation-M inhibits progression of periodontal tissue destruction.

Persson, G. R.; Engel, D.; Whitney, C.; Darveau, R.; Weinberg, A.; Brunsvold, M.; Page, R. C.

1994-01-01

82

Fimbria-mediated coaggregation between human oral anaerobes Treponema medium and Porphyromonas gingivalis.  

Science.gov (United States)

Bacterial binding phenomena among different bacterial genera or species play an important role in bacterial colonization in a mixed microbiota such as in the human oral cavity. The coaggregation reaction between two gram-negative anaerobes, Treponema medium and Porphyromonas gingivalis, was characterized using fimbria-deficient mutants of P. gingivalis and specific antisera against purified fimbriae and bacterial whole cells. T. medium ATCC 700273 strongly coaggregated with fimbriate P. gingivalis strains ATCC 33277 and 381, but not with afimbriate strains including transposon-induced fimbria-deficient mutants and KDP98 as a fimA-disrupted mutant of P. gingivalis ATCC 33277. In the P. gingivalis-T. medium coaggregation assay, the presence of rabbit antiserum against the purified fimbriae or the whole cells of P. gingivalis ATCC 33277 produced different "aggregates" consisting predominantly of P. gingivalis cells with few spirochetes, but both preimmune serum and the antiserum against the afimbriate KDP98 cells did not inhibit the coaggregation reaction. Heated P. gingivalis cells lost their ability to bind both heated and unheated T. medium cells. This T. medium-P. gingivalis coaggregation reaction was inhibited by a cysteine proteinase inhibitor, leupeptin, and also by arginine and lysine, but not by EDTA or sugars including lactose. A binding assay on nitrocellulose membranes and immunoelectron microscopy demonstrated that a heat-stable 37 kDa surface protein on the T. medium cell attached to the P. gingivalis fimbriae. PMID:10553676

Umemoto, T; Yoshimura, F; Kureshiro, H; Hayashi, J; Noguchi, T; Ogawa, T

1999-01-01

83

OxyR Activation in Porphyromonas gingivalis in Response to a Hemin-Limited Environment  

OpenAIRE

Porphyromonas gingivalis is a Gram-negative obligately anaerobic bacterium associated with several forms of periodontal disease, most closely with chronic periodontitis. Previous studies demonstrated that OxyR plays an important role in the aerotolerance of P. gingivalis by upregulating the expression of oxidative-stress genes. Increases in oxygen tension and in H2O2 both induce activation of OxyR. It is also known that P. gingivalis requires hemin as an iron source for its growth. In this st...

Xie, Hua; Zheng, Cunge

2012-01-01

84

Distribution and Molecular Characterization of Porphyromonas gingivalis Carrying a New Type of fimA Gene  

OpenAIRE

Fimbriae of Porphyromonas gingivalis are filamentous appendages on the cell surface and are thought to be one of the virulence factors. The fimA gene encoding the subunit protein of fimbriae, fimbrillin (FimA), was classified into four typeable variants (types I to IV). We previously examined the distribution of P. gingivalis in terms of fimA genotypes in periodontitis patients using a fimA type-specific PCR assay. However, some patients harbored P. gingivalis with untypeable fimA. In this st...

Nakagawa, Ichiro; Amano, Atsuo; Kimura, Richard K.; Nakamura, Takayuki; Kawabata, Shigetada; Hamada, Shigeyuki

2000-01-01

85

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

OpenAIRE

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 ligame...

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

NOD2 contributes to Porphyromonas gingivalis-induced bone resorption.  

Science.gov (United States)

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

87

Porphyromonas gingivalis: keeping the pathos out of the biont  

Directory of Open Access Journals (Sweden)

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

88

Suppression of inflammatory responses of human gingival fibroblasts by gingipains from Porphyromonas gingivalis.  

Science.gov (United States)

The interaction between human gingival fibroblasts (HGFs) and Porphyromonas gingivalis plays an important role in the development and progression of periodontitis. Porphyromonas gingivalis possesses several virulence factors, including cysteine proteases, the arginine-specific (Rgp) and lysine-specific (Kgp) gingipains. Studying the mechanisms that P. gingivalis, and its derived virulence, use to propagate and interact with host cells will increase the understanding of the development and progression of periodontitis. In this study, we aimed to elucidate how P. gingivalis influences the inflammatory events in HGFs regarding transforming growth factor-?1 (TGF-?1 ), CXCL8, secretory leucocyte protease inhibitor (SLPI), c-Jun and indoleamine 2,3-dioxygenase (IDO). HGFs were inoculated for 6 and 24 h with the wild-type strains ATCC 33277 and W50, two gingipain-mutants of W50 and heat-killed ATCC 33277. The P. gingivalis regulated CXCL8 and TGF-?1 in HGFs, and the kgp mutant gave significantly higher immune response with increased CXCL8 (P < 0.001) and low levels of TGF-?1 . We show that HGFs express and secrete SLPI, which was significantly suppressed by P. gingivalis (P < 0.05). This suggests that by antagonizing SLPI, P. gingivalis contributes to the tissue destruction associated with periodontitis. Furthermore, we found that P. gingivalis inhibits the expression of the antimicrobial IDO, as well as upregulating c-Jun (P < 0.05). In conclusion, P. gingivalis both triggers and suppresses the immune response in HGFs. Consequently, we suggest that the pathogenic effects of P. gingivalis, and especially the activity of the gingipains on the inflammatory and immune response of HGFs, are crucial in periodontitis. PMID:25055828

Palm, E; Khalaf, H; Bengtsson, T

2015-02-01

89

Genetic Diversity of Porphyromonas gingivalis Isolates Recovered from Single “Refractory” Periodontitis Sites?  

OpenAIRE

Multilocus sequence typing and fimA genotyping were performed on Porphyromonas gingivalis isolates from 15 subjects with “refractory” periodontitis. Several sequence types were detected for most individual pockets. The variation indicated recombination at the recA and pepO genes. The prevalence of fimA genotypes II and IV confirmed their association with periodontitis.

Enersen, Morten; Olsen, Ingar; Caugant, Dominique A.

2008-01-01

90

Altered gingipain maturation in vimA- and vimE-defective isogenic mutants of Porphyromonas gingivalis.  

Science.gov (United States)

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 vimE-defective mutant. In contrast, the membrane of P. gingivalis FLL92, the vimA-defective mutant, demonstrated immunoreactivity only with RgpB antibodies. With mass spectrometry or Western blots, full-length RgpA and RgpB were identified from extracellular fractions. In similar extracellular fractions from P. gingivalis FLL92 and FLL93, purified RgpB activated only arginine-specific activity. In addition, the lipopolysaccharide profiles of the vimA and vimE mutants were truncated in comparison to that of W83. While glycosylated proteins were detected in the membrane and extracellular fractions from the vimA- and vimE-defective mutants, a monoclonal antibody (1B5) that reacts with specific sugar moieties of the P. gingivalis cell surface polysaccharide and membrane-associated Rgp gingipain showed no immunoreactivity with these fractions. Taken together, these results indicate a possible defect in sugar biogenesis in both the vimA- and vimE-defective mutants. These modulating genes play a role in the secretion, processing, and/or anchorage of gingipains on the cell surface. PMID:15731033

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

2005-03-01

91

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

Science.gov (United States)

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

92

Disruption of heterotypic community development by Porphyromonas gingivalis with small molecule inhibitors.  

Science.gov (United States)

Porphyromonas gingivalis is one of the main etiological organisms in periodontal disease. On oral surfaces P. gingivalis is a component of multispecies biofilm communities and can modify the pathogenic potential of the community as a whole. Accumulation of P. gingivalis in communities is facilitated by interspecies binding and communication with the antecedent colonizer Streptococcus gordonii. In this study we screened a library of small molecules to identify structures that could serve as lead compounds for the development of inhibitors of P. gingivalis community development. Three small molecules were identified that effectively inhibited accumulation of P. gingivalis on a substratum of S. gordonii. The structures of the small molecules are derived from the marine alkaloids oroidin and bromoageliferin and contain a 2-aminoimidazole or 2-aminobenzimidazole moiety. The most active compounds reduced expression of mfa1 and fimA in P. gingivalis, genes encoding the minor and major fimbrial subunits, respectively. These fimbrial adhesins are necessary for P. gingivalis co-adhesion with S. gordonii. These results demonstrate the potential for a small molecular inhibitor-based approach to the prevention of diseases associated with P. gingivalis. PMID:24899524

Wright, C J; Wu, H; Melander, R J; Melander, C; Lamont, R J

2014-10-01

93

Fusobacterium nucleatum supports the growth of Porphyromonas gingivalis in oxygenated and carbon-dioxide-depleted environments.  

Science.gov (United States)

The authors compared the differences in tolerance to oxygen of the anaerobic periodontopathic bacteria Fusobacterium nucleatum and Porphyromonas gingivalis, and explored the possibility that F. nucleatum might be able to support the growth of P. gingivalis in aerated and CO2-depleted environments. Both micro-organisms were grown as monocultures and in co-culture in the presence and absence of CO2 and under different aerated conditions using a continuous culture system. At steady state, viable counts were performed and the activities of the enzymes superoxide dismutase and NADH oxidase/peroxidase were assayed in P. gingivalis. In co-culture, F. nucleatum was able to support the growth of P. gingivalis in aerated and CO2-depleted environments in which P. gingivalis, as a monoculture, was not able to survive. F. nucleatum not only appeared to have a much higher tolerance to oxygen than P. gingivalis, but a significant increase in its numbers occurred under moderately oxygenated conditions. F. nucleatum might have an additional indirect role in dental plaque maturation, contributing to the reducing conditions necessary for the survival of P. gingivalis and possibly other anaerobes less tolerant to oxygen. Additionally, F. nucleatum is able to generate a capnophilic environment essential for the growth of P. gingivalis. PMID:11832510

Diaz, P I; Zilm, P S; Rogers, A H

2002-02-01

94

Pyocycanin, a Contributory Factor in Haem Acquisition and Virulence Enhancement of Porphyromonas gingivalis in the Lung  

Science.gov (United States)

Several recent studies show that the lungs infected with Pseudomonas aeruginosa are often co-colonised by oral bacteria including black-pigmenting anaerobic (BPA) Porphyromonas species. The BPAs have an absolute haem requirement and their presence in the infected lung indicates that sufficient haem, a virulence up-regulator in BPAs, must be present to support growth. Haemoglobin from micro-bleeds occurring during infection is the most likely source of haem in the lung. Porphyromonas gingivalis displays a novel haem acquisition paradigm whereby haemoglobin must be firstly oxidised to methaemoglobin, facilitating haem release, either by gingipain proteolysis or capture via the haem-binding haemophore HmuY. P. aeruginosa produces the blue phenazine redox compound, pyocyanin. Since phenazines can oxidise haemoglobin, it follows that pyocyanin may also facilitate haem acquisition by promoting methaemoglobin production. Here we show that pyocyanin at concentrations found in the CF lung during P. aeruginosa infections rapidly oxidises oxyhaemoglobin in a dose-dependent manner. We demonstrate that methaemoglobin formed by pyocyanin is also susceptible to proteolysis by P. gingivalis Kgp gingipain and neutrophil elastase, thus releasing haem. Importantly, co-incubation of oxyhaemoglobin with pyocyanin facilitates haem pickup from the resulting methemoglobin by the P. gingivalis HmuY haemophore. Mice intra-tracheally challenged with viable P. gingivalis cells plus pyocyanin displayed increased mortality compared to those administered P. gingivalis alone. Pyocyanin significantly elevated both methaemoglobin and total haem levels in homogenates of mouse lungs and increased the level of arginine-specific gingipain activity from mice inoculated with viable P. gingivalis cells plus pyocyanin compared with mice inoculated with P. gingivalis only. These findings indicate that pyocyanin, by promoting haem availability through methaemoglobin formation and stimulating of gingipain production, may contribute to virulence of P. gingivalis and disease severity when co-infecting with P. aeruginosa in the lung. PMID:25706529

Benedyk, Malgorzata; Byrne, Dominic P.; Glowczyk, Izabela; Potempa, Jan; Olczak, Mariusz; Olczak, Teresa; Smalley, John W.

2015-01-01

95

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

OpenAIRE

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

96

Strong cytotoxicity to human gingival fibroblasts by Porphyromonas gingivalis ATCC 33277.  

Science.gov (United States)

The aim of the present study was to analyze the cytotoxicity of some bacterial species associated with periodontal diseases. The specificity of cytotoxicity was estimated against cells of various origin and from different individuals. The reference bacteria were Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Fusobacterium nucleatum. These bacteria were cultured for 24 h in liquid media and the supernatants were used in cytotoxicity assays. The target cells used were human gingival fibroblasts (GF), dermal fibroblasts (K4), gingival epithelial cells (E) and HeLa-cells (HeLa). These cells were exposed at 4 h or 24 h, respectively, to various concentrations of culture supernatants from the selected bacteria. The influence on the viability and metabolism of the cells were estimated quantitatively as increase in neutral red uptake and lactic acid production. Growth medium supernatants of P. gingivalis 33277 were strongly cytotoxic to gingival fibroblasts after 24 h incubation, compared to supernatants of P. gingivalis 381 or W 50, A. actinomycetemcomitans or F. nucleatum cultures. The toxic effect of P. gingivalis 33277 decreased drastically after heat inactivation, which indicates effects of proteins. By adding anti-sera the cytotoxicity of P. gingivalis 33277 could be dose dependently neutralized, which was not the case when supernatants of A. actino-mycetemcomitans was tested. Target cells of epithelial origin did not show increased cytotoxicity to P. gingivalis 33277. The results of the present study strengthen the hypothesis that P. gingivalis remains as a suspect causative key component in periodontal diseases. PMID:8915950

Johansson, A; Bergenholtz, A; Holm, S E

1996-10-01

97

Inhibition of Porphyromonas gingivalis adhesion to Streptococcus gordonii by human submandibular-sublingual saliva.  

Science.gov (United States)

Porphyromonas gingivalis W50 adheres in vitro to biofilms of Streptococcus gordonii G9B. This phenomenon is believed to facilitate the initial colonization of the oral cavity by P. gingivalis and to contribute to the maturation of dental plaque. In this report, we describe the modulating effects of human submandibular-sublingual saliva (HSMSL) on this in vitro model of intergeneric bacterial adhesion (coaggregation). HSMSL inhibited P. gingivalis adhesion to S. gordonii by 50% at a concentration of 57 micrograms of protein per ml. Maximum inhibitory activity was associated with a 43-kDa protein obtained by sequential Sephadex G200 gel filtration and CM52 ion-exchange chromatography of HSMSL. Pools of other column fractions of HSMSL showed no effect or were slightly stimulatory for bacterial adhesion. The binding of radioiodinated column fractions containing the 43-kDa protein by P. gingivalis was accompanied by their rapid enzymatic degradation. Treating P. gingivalis at 60 degrees C for 30 min or with protease inhibitors (phenylmethylsulfonyl fluoride and sodium iodoacetate) reduced adherence to streptococcal biofilms. These treatments did not prevent P. gingivalis from binding soluble HSMSL saliva components, although subsequent proteolysis was nearly eliminated. These observations indicate that surface-associated proteases of P. gingivalis, either independently or in concert with adjacent surface adhesins, interact with surfaces of oral streptococci to facilitate interbacterial adhesion. The adhesion-blocking properties of HSMSL, particularly the 43-kDa protein, may represent an important host defense mechanism in the oral cavity. PMID:1319402

Stinson, M W; Haraszthy, G G; Zhang, X L; Levine, M J

1992-07-01

98

Porphyromonas gingivalis infection reduces regulatory T cells in infected atherosclerosis patients.  

Science.gov (United States)

Increasing evidence has shown periodontal pathogen Porphyromonas gingivalis (P.gingivalis) infection contributes to atherosclerosis (AS) progression. P.gingivalis fimbriae act as an important virulence factor in AS. Regulatory T cells (Tregs) may play a crucial role in autoimmune response during this process. However, whether P.gingivalis infection is associated with Tregs dysregulation during AS is still unknown and the prevalence of different P.gingivalis FimA genotypes during this process is unclear. Here we analyzed the distribution of Tregs and in P.gingivalis-infected atherosclerotic patients to reveal the relationship between P.gingivalis infection and Tregs reduction/dysfunction and to elucidate their role in periodontitis-AS interaction. FimA genotype was also examined to determine the prevalence of fimbriae. Our results showed that P.gingivalis infection reduced Tregs in atherosclerotic patients compared with non-atherosclerotic patients and health controls. Concentration of TGF-?1, which plays an important role in the development of Tregs, also decreased in P.gingivalis infected patients. Furthermore, type II FimA seems to show higher prevalence than the other five detected types. The population of Tregs further decreased in patients with type II FimA compared with the other types. P.gingivlias FimA genotype II was the dominant type associated with decreased Treg population. These results indicate that P.gingivalis infection may be associated with Tregs dysregulation in AS; type II FimA may be a predominant genotype in this process. PMID:24466164

Yang, Jie; Wu, Juan; Liu, Yu; Huang, Jin; Lu, Zhipin; Xie, Liping; Sun, Weibin; Ji, Yong

2014-01-01

99

Fimbriae and the hemagglutinating adhesin HA-Ag2 mediate adhesion of Porphyromonas gingivalis to epithelial cells.  

OpenAIRE

The mechanisms by which Porphyromonas gingivalis, a gram-negative anaerobic bacterium, is pathogenic for the periodontium remain largely hypothetical. Invasion of host tissues by P. gingivalis is believed to require adhesion of the bacterium to host cells. The aim of this study was to use monoclonal antibodies (MAbs) to characterize the bacterial cell surface component(s) acting as a ligand binding to a receptor on epithelial cells. Surface antigens of P. gingivalis ATCC 33277 were obtained a...

Du, L.; Pellen-mussi, P.; Chandad, F.; Mouton, C.; Bonnaure-mallet, M.

1997-01-01

100

Formation of Methyl Mercaptan from l-Methionine by Porphyromonas gingivalis  

OpenAIRE

Methyl mercaptan production by oral bacteria is thought to be one of the main causes of oral malodor. We examined the ability of periodontopathic Porphyromonas gingivalis to produce methyl mercaptan from l-methionine and found that the invasive strains W83 and W50 produced large amounts of methyl mercaptan. We cloned and sequenced the mgl gene encoding l-methionine-?-deamino-?-mercaptomethane-lyase (METase) from P. gingivalis W83. The structural mgl gene consisted of 1,200 bp and encoded a ...

Yoshimura, Mamiko; Nakano, Yoshio; Yamashita, Yoshihisa; Oho, Takahiko; Saito, Toshiyuki; Koga, Toshihiko

2000-01-01

101

Tetratricopeptide Repeat Protein-Associated Proteins Contribute to the Virulence of Porphyromonas gingivalis? †  

OpenAIRE

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 e...

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

2010-01-01

102

Transformation and Expression of a Cloned fimA Gene in Porphyromonas gingivalis  

OpenAIRE

The Porphyromonas gingivalis fimbria is an important virulence factor involved in the adherence and colonization of the organism in the oral cavity. In this study, we transformed this organism with a gene, fimA381, encoding the fimbrial subunit of P. gingivalis 381 (fimbrillin) by using the host-vector system that we developed previously and examined expression of the cloned fimA381 gene. The recombinant plasmid pYHF2 was constructed by ligating a fragment containing the fimA381 gene into the...

Takahashi, Yusuke; Kato, Daisuke; Hamada, Nobushiro; Yoshimoto, Hisashi; Umemoto, Toshio

1999-01-01

103

Identification of Porphyromonas gingivalis components that mediate its interactions with fibronectin.  

OpenAIRE

Porphyromonas (Bacteroides) gingivalis W12 binds and degrades human plasma fibronectin. In the presence of the protease inhibitor N-alpha-p-tosyl-L-lysyl chloromethyl ketone, P. gingivalis cells accumulated substantial amounts of 125I-fibronectin as a function of incubation time. Fibronectin binding was specific, reversible, and saturable. The Kd for the reaction was estimated to be on the order of 100 nM, and there was an average of 3.5 x 10(3) fibronectin binding sites per cell. Unlabeled f...

Lantz, M. S.; Allen, R. D.; Duck, L. W.; Blume, J. L.; Switalski, L. M.; Hook, M.

1991-01-01

104

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

OpenAIRE

Recombinant VimA protein can interact with the gingipains and several other proteins that may play a role in its biogenesis in Porphyromonas gingivalis. In silico analysis of PG2096, a hypothetical protein that was shown to interact with VimA, suggests that it may have environmental stress resistance properties. To further evaluate the role(s) of PG2096, the predicted open reading frame was PCR amplified from P. gingivalis W83 and insertionally inactivated using the ermF-ermAM antibiotic-resi...

Vanterpool, E.; Aruni, A. Wilson; Roy, F.; Fletcher, H. M.

2010-01-01

105

VimA mediates multiple functions that control virulence in Porphyromonas gingivalis  

OpenAIRE

Porphyromonas gingivalis, a black-pigmented, gram-negative anaerobe, is an important etiological agent of periodontal disease. Its ability to survive in the periodontal pocket and orchestrate the microbial/host activities that can lead to disease suggest that P. gingivalis possesses a complex regulatory network involving transcriptional and post-transcriptional mechanisms. The vimA (virulence modulating) gene is part of the 6.15-kb bcp-recA-vimA-vimE-vimF-aroG locus and plays a role in oxidat...

Aruni, A. Wilson; Robles, A.; Fletcher, H. M.

2012-01-01

106

Highly specific protease-based approach for detection of porphyromonas gingivalis in diagnosis of periodontitis.  

Science.gov (United States)

Porphyromonas gingivalis is associated with the development of periodontitis. Here we describe the development of a highly specific protease-based diagnostic method for the detection of P. gingivalis in gingival crevicular fluid. Screening of a proteolytic peptide substrate library, including fluorogenic dipeptides that contain d-amino acids, led to the discovery of five P. gingivalis-specific substrates. Due to the presence of lysine and arginine residues in these substrates, it was hypothesized that the cleavage was mediated by the gingipains, a group of P. gingivalis-specific proteases. This hypothesis was confirmed by the observation that P. gingivalis gingipain knockout strains demonstrated clearly impaired substrate cleavage efficacy. Further, proteolytic activity on the substrates was increased by the addition of the gingipain stimulators dithiothreitol and l-cysteine and decreased by the inhibitors leupeptin and N-ethylmaleimide. Screening of saliva and gingival crevicular fluid of periodontitis patients and healthy controls showed the potential of the substrates to diagnose the presence of P. gingivalis proteases. By using paper points, a sensitivity of approximately 10(5) CFU/ml was achieved. P. gingivalis-reactive substrates fully composed of l-amino acids and Bz-l-Arg-NHPhNO(2) showed a relatively low specificity (44 to 85%). However, the five P. gingivalis-specific substrates that each contained a single d-amino acid showed high specificity (96 to 100%). This observation underlines the importance of the presence of d-amino acids in substrates used for the detection of bacterial proteases. We envisage that these substrates may improve the specificity of the current enzyme-based diagnosis of periodontitis associated with P. gingivalis. PMID:22075590

Kaman, Wendy E; Galassi, Fabiano; de Soet, Johannes J; Bizzarro, Sergio; Loos, Bruno G; Veerman, Enno C I; van Belkum, Alex; Hays, John P; Bikker, Floris J

2012-01-01

107

Inducible expression of a Porphyromonas gingivalis W83 membrane-associated protease.  

OpenAIRE

The Tpr protease of Porphyromonas gingivalis W83 is a membrane-associated enzyme capable of hydrolyzing a chromogenic bacterial collagenase substrate. An isogenic mutant lacking a functional tpr gene had a greatly reduced ability to hydrolyze the collagenase substrate. Activity was restored to the tpr mutant by introducing a shuttle plasmid containing the tpr gene. Expression of the gene is induced by nutrient limitation, as shown by enzymatic and Northern analyses.

Park, Y.; Lu, B.; Mazur, C.; Mcbride, B. C.

1997-01-01

108

Porphyromonas gingivalis: a clonal pathogen?: Diversities in housekeeping genes and the major fimbriae gene  

OpenAIRE

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 dive...

Enersen, Morten

2011-01-01

109

Functional differences of Porphyromonas gingivalis Fimbriae in determining periodontal disease pathogenesis: a literature review  

OpenAIRE

Porphyromonas gingivalis is implicated in chronic and aggressive periodontitis. This bacterium has numerous virulence factors and one is the Fimbriae, which is quite important for bacterial colonization. Fimbriae are appendices that anchor to the bacterial wall and are comprised of the protein FimBriline encoded by the FimA gene. Thus far, six genotypes have been identified, FimA I to V and Ib. Genotypes II and IV are associated with periodontal disease, while genotype I is related to gingiva...

Moreno, Sandra; Contreras, Adolfo

2013-01-01

110

Differential Activation of Human Gingival Epithelial Cells and Monocytes by Porphyromonas gingivalis Fimbriae?  

OpenAIRE

Humans develop periodontitis in response to challenge by microbial dental plaque. Inflammation begins after perturbation of gingival epithelial cells by subgingival bacteria interacting through pattern-recognition receptors, including the Toll-like receptors (TLR). Porphyromonas gingivalis is a major periodontopathogen that interacts with epithelial cells through its cell surface fimbriae (FimA), leading to colonization and/or invasion. Previous work by our group has established membrane CD14...

Eskan, Mehmet A.; Hajishengallis, George; Kinane, Denis F.

2006-01-01

111

Divergence of the systemic immune response following oral infection with distinct strains of Porphyromonas gingivalis  

OpenAIRE

Periodontitis is a polymicrobial oral infection characterized by the destruction of tooth-supporting structures that can be linked to systemic diseases such as cardiovascular disease, diabetes or rheumatoid arthritis. Porphyromonas gingivalis, a bacterium implicated in the etiology of periodontitis, has shown variation in inducing T-cell responses among different strains. Therefore, in this study we investigated the strain-specific immune response using a murine experimental model of periodon...

Marchesan, J. T.; Morelli, T.; Lundy, S. K.; Jiao, Y.; Lim, S.; Inohara, N.; Nunez, G.; Fox, D. A.; Giannobile, W. V.

2012-01-01

112

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

OpenAIRE

A-type cranberry proanthocyanidins (AC-PACs) have recently been reported to be beneficial for human health, especially urinary tract health. The effect of these proanthocyanidins on periodontitis, a destructive disease of tooth-supporting tissues, needs to be investigated. The purpose of this study was to investigate the effects of AC-PACs on various virulence determinants of Porphyromonas gingivalis as well as on the inflammatory response of oral epithelial cells stimulated by this periodont...

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

2010-01-01

113

Proteomic and Transcriptional Analysis of Interaction between Oral Microbiota Porphyromonas gingivalis and Streptococcus oralis.  

Science.gov (United States)

Porphyromonas gingivalis, a major periodontal pathogen, forms biofilm with other oral bacteria such as streptococci. Here, by using shotgun proteomics, we examined the molecular basis of mixed-biofilm formation by P. gingivalis with Streptococcus oralis. We identified a total of 593 bacterial proteins in the biofilm. Compared to the expression profile in the P. gingivalis monobiofilm, the expression of three proteins was induced and that of 31 proteins was suppressed in the mixed biofilm. Additionally, the expression of two S. oralis proteins was increased, while that of two proteins was decreased in the mixed biofilm, as compared to its monotypic profile. mRNA expression analysis of selected genes using a quantitative reverse transcription polymerase chain reaction confirmed the proteomics data, which included overexpression of P. gingivalis FimA and S. oralis glyceraldehyde-3-phosphate dehydrogenase in association with the biofilm. The results also indicated that S. oralis regulates the transcriptional activity of P. gingivalis luxS to influence autoinducer-2-dependent signaling. These findings suggest that several functional molecules are involved in biofilm formation between P. gingivalis and S. oralis. PMID:25341202

Maeda, Kazuhiko; Nagata, Hideki; Ojima, Miki; Amano, Atsuo

2015-01-01

114

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

Directory of Open Access Journals (Sweden)

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

115

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

Science.gov (United States)

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 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

116

Fimbriae-mediated outer membrane vesicle production and invasion of Porphyromonas gingivalis.  

Science.gov (United States)

Porphyromonas gingivalis is a keystone periopathogen that plays an essential role in the progress of periodontitis. Like other gram-negative bacteria, the ability of P. gingivalis to produce outer membrane vesicles is a strategy used to interact with, and survive within its biological niches. Here we compared the protein components associated with vesicles derived from a fimbriated strain (33277) and an afimbriated strain (W83) of P. gingivalis using proteomic analyses. Some well-known virulence factors were identified in vesicles from both strains, such as gingipains and hemagglutinin. In contrast, FimC, FimD, and FimE, minor components of long fimbriae were found exclusively in 33277 vesicles, while proteins with a tetratricopeptide repeat (TPR) domain were unique to W83 vesicles. We found that significantly more 33277 than W83 vesicles were internalized into human oral keratinocytes and gingival fibroblasts. Interestingly, FimA, a well-known adhesin responsible for the attachment and invasion of P. gingivalis into host cells, was not essential for the invasive capabilities of P. gingivalis vesicles. Rather minor components of long fimbriae were required for an efficient invasive activity of vesicles. The most striking finding was that P. gingivalis strains lacking or having a reduced FimA expression showed a significant reduction in vesiculation. These results suggest that production and pathogenicity of P. gingivalis vesicles may largely depend on expression of the fim locus, and that the integration of vesicle production and pathogenicity with fimbrial expression may allow P. gingivalis to confer upon itself certain functional advantages. PMID:25524808

Mantri, Chinmay K; Chen, Chin-Ho; Dong, Xinhong; Goodwin, Jeffery Shawn; Pratap, Siddharth; Paromov, Victor; Xie, Hua

2015-02-01

117

Fimbriated Porphyromonas gingivalis Is More Efficient than Fimbria-Deficient P. gingivalis in Entering Human Dendritic Cells In Vitro and Induces an Inflammatory Th1 Effector Response  

OpenAIRE

Porphyromonas gingivalis is a fimbriated mucosal pathogen implicated in chronic periodontitis (CP). The fimbriae are required for invasion of the gingival mucosa and for induction of CP in animal models of periodontitis. CP is associated with infection of immature dendritic cells (DCs) by P. gingivalis in situ and with increased numbers of dermal DCs (DDCs) and mature DCs in the lamina propria. The role of fimbriae in gaining entry into human DCs and how this modulates the inflammatory and ef...

Jotwani, Ravi; Cutler, Christopher W.

2004-01-01

118

Identification of a diguanylate cyclase and its role in Porphyromonas gingivalis virulence.  

Science.gov (United States)

Porphyromonas gingivalis is a Gram-negative obligate anaerobic bacterium and is considered a keystone pathogen in the initiation of periodontitis, one of the most widespread infectious diseases. Bacterial bis-(3'-5') cyclic GMP (cyclic di-GMP [c-di-GMP]) serves as a second messenger and is involved in modulating virulence factors in numerous bacteria. However, the role of this second messenger has not been investigated in P. gingivalis, mainly due to a lack of an annotation regarding diguanylate cyclases (DGCs) in this bacterium. Using bioinformatics tools, we found a protein, PGN_1932, containing a GGDEF domain. A deletion mutation in the pgn_1932 gene had a significant effect on the intracellular c-di-GMP level in P. gingivalis. Genetic analysis showed that expression of the fimA and rgpA genes, encoding the major protein subunit of fimbriae and an arginine-specific proteinase, respectively, was downregulated in the pgn_1932 mutant. Correspondingly, FimA protein production and the fimbrial display on the mutant were significantly reduced. Mutations in the pgn_1932 gene also had a significant impact on the adhesive and invasive capabilities of P. gingivalis, which are required for its pathogenicity. These findings provide evidence that the PGN_1932 protein is both responsible for synthesizing c-di-GMP and involved in biofilm formation and host cell invasion by P. gingivalis by controlling the expression and biosynthesis of FimA. PMID:24733094

Chaudhuri, Swarnava; Pratap, Siddharth; Paromov, Victor; Li, Zhijun; Mantri, Chinmay K; Xie, Hua

2014-07-01

119

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

Science.gov (United States)

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

120

Role of vimA in cell surface biogenesis in Porphyromonas gingivalis.  

Science.gov (United States)

The Porphyromonas gingivalis vimA gene has been previously shown to play a significant role in the biogenesis of gingipains. Further, in P. gingivalis FLL92, a vimA-defective mutant, there was increased auto-aggregation, suggesting alteration in membrane surface proteins. In order to determine the role of the VimA protein in cell surface biogenesis, the surface morphology of P. gingivalis FLL92 was further characterized. Transmission electron microscopy demonstrated abundant fimbrial appendages and a less well defined and irregular capsule in FLL92 compared with the wild-type. In addition, atomic force microscopy showed that the wild-type had a smoother surface compared with FLL92. Western blot analysis using anti-FimA antibodies showed a 41 kDa immunoreactive protein band in P. gingivalis FLL92 which was missing in the wild-type P. gingivalis W83 strain. There was increased sensitivity to globomycin and vancomycin in FLL92 compared with the wild-type. Outer membrane fractions from FLL92 had a modified lectin-binding profile. Furthermore, in contrast with the wild-type strain, nine proteins were missing from the outer membrane fraction of FLL92, while 20 proteins present in that fraction from FLL92 were missing in the wild-type strain. Taken together, these results suggest that the VimA protein affects capsular synthesis and fimbrial phenotypic expression, and plays a role in the glycosylation and anchorage of several surface proteins. PMID:20378652

Osbourne, Devon O; Aruni, Wilson; Roy, Francis; Perry, Christopher; Sandberg, Lawrence; Muthiah, Arun; Fletcher, Hansel M

2010-07-01

121

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

Science.gov (United States)

In several micro-organisms, HtrA, a serine periplasmic protease, is considered an important virulence factor that plays a regulatory role in oxidative and temperature stress. The authors have previously shown that the vimA gene product is an important virulence regulator in Porphyromonas gingivalis. Further, purified recombinant VimA physically interacted with the major gingipains and the HtrA from P. gingivalis. To further evaluate a role for HtrA in the pathogenicity of this organism, a 1.5 kb fragment containing the htrA gene was PCR-amplified from the chromosomal DNA of P. gingivalis W83. This gene was insertionally inactivated using the ermF-ermAM antibiotic-resistance cassette and used to create an htrA-deficient mutant by allelic exchange. In one randomly chosen isogenic mutant designated P. gingivalis FLL203, there was increased sensitivity to hydrogen peroxide. Growth of this mutant at an elevated temperature was more inhibited compared to the wild-type. Further, in contrast to the wild-type, there was a significant decrease in Arg-gingipain activity after heat shock in FLL203. However, the gingipain activity in the mutant returned to normal levels after a further 30 min incubation at room temperature. Collectively, these data suggest that HtrA may play a similar role in oxidative and temperature stress in P. gingivalis as observed in other organisms. PMID:17074908

Roy, F; Vanterpool, E; Fletcher, H M

2006-11-01

122

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

123

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 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

124

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

Science.gov (United States)

Porphyromonas gingivalis, is a Gram-negative obligate oral anaerobic bacterium highly implicated in periodontal disease, the most prevalent chronic inflammatory disease, but recent evidence also indicates a potential contribution to systemic inflammation. The Triggering Receptor Expressed on Myeloid cells 1 (TREM-1) is a cell surface receptor of the immunoglobulin superfamily, which, along with its adaptor signalling molecule DAP12, is involved in immune response to bacterial and fungal infections, particularly by amplifying the production of pro-inflammatory cytokines by the host. The aim of the present study was to investigate the effect of P. gingivalis on the expression of the TREM-1/DAP12 pathway, as well as its engagement in pro-inflammatory cytokine production, by the myelomonocytic cell line MonoMac-6. P. gingivalis enhanced TREM-1 gene expression by the cells, concomitantly to an increase of soluble TREM-1 secretion. Engagement of TREM-1, by introducing anti-TREM-1 to the experimental system, resulted in further potentiation of the pro-inflammatory responses to P. gingivalis, as evaluated by a further enhancement of interleukin (IL)-1? and IL-6 secretion. On the contrary, the synthetic TREM-1 antagonist LP17 reduced the P. gingivalis-induced IL-1? and IL-6 secretion by approximately 50%. In conclusion, the putative periodontal pathogen P. gingivalis can positively regulate the expression of the TREM-1/DAP12 pathway in monocytic cells. Moreover, engagement of TREM-1 can further potentiate the pro-inflammatory responses to P. gingivalis infection. This effect may contribute not only to the pathogenesis of inflammatory periodontal disease, but also to the enhancement of systemic inflammation. PMID:21967868

Bostanci, Nagihan; Thurnheer, Thomas; Belibasakis, Georgios N

2011-10-01

125

The Porphyromonas gingivalis ferric uptake regulator orthologue binds hemin and regulates hemin-responsive biofilm development.  

Science.gov (United States)

Porphyromonas gingivalis is a Gram-negative pathogen associated with the biofilm-mediated disease chronic periodontitis. P. gingivalis biofilm formation is dependent on environmental heme for which P. gingivalis has an obligate requirement as it is unable to synthesize protoporphyrin IX de novo, hence P. gingivalis transports iron and heme liberated from the human host. Homeostasis of a variety of transition metal ions is often mediated in Gram-negative bacteria at the transcriptional level by members of the Ferric Uptake Regulator (Fur) superfamily. P. gingivalis has a single predicted Fur superfamily orthologue which we have designated Har (heme associated regulator). Recombinant Har formed dimers in the presence of Zn2+ and bound one hemin molecule per monomer with high affinity (Kd of 0.23 µM). The binding of hemin resulted in conformational changes of Zn(II)Har and residue 97Cys was involved in hemin binding as part of a predicted -97C-98P-99L- hemin binding motif. The expression of 35 genes was down-regulated and 9 up-regulated in a Har mutant (ECR455) relative to wild-type. Twenty six of the down-regulated genes were previously found to be up-regulated in P. gingivalis grown as a biofilm and 11 were up-regulated under hemin limitation. A truncated Zn(II)Har bound the promoter region of dnaA (PGN_0001), one of the up-regulated genes in the ECR455 mutant. This binding decreased as hemin concentration increased which was consistent with gene expression being regulated by hemin availability. ECR455 formed significantly less biofilm than the wild-type and unlike wild-type biofilm formation was independent of hemin availability. P. gingivalis possesses a hemin-binding Fur orthologue that regulates hemin-dependent biofilm formation. PMID:25375181

Butler, Catherine A; Dashper, Stuart G; Zhang, Lianyi; Seers, Christine A; Mitchell, Helen L; Catmull, Deanne V; Glew, Michelle D; Heath, Jacqueline E; Tan, Yan; Khan, Hasnah S G; Reynolds, Eric C

2014-01-01

126

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

Science.gov (United States)

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

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

2013-01-01

127

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.

2013-01-01

128

Invasion of Epithelial Cells and Proteolysis of Cellular Focal Adhesion Components by Distinct Types of Porphyromonas gingivalis Fimbriae  

OpenAIRE

Porphyromonas gingivalis fimbriae are classified into six types (types I to V and Ib) based on the fimA genes encoding FimA (a subunit of fimbriae), and they play a critical role in bacterial interactions with host tissues. In this study, we compared the efficiencies of P. gingivalis strains with distinct types of fimbriae for invasion of epithelial cells and for degradation of cellular focal adhesion components, paxillin, and focal adhesion kinase (FAK). Six representative strains with the d...

Nakagawa, Ichiro; Inaba, Hiroaki; Yamamura, Taihei; Kato, Takahiro; Kawai, Shinji; Ooshima, Takashi; Amano, Atsuo

2006-01-01

129

Importance of TLR2 in Early Innate Immune Response to Acute Pulmonary Infection with Porphyromonas gingivalis in Mice 1  

OpenAIRE

The periodontal pathogen Porphyromonas gingivalis is implicated in certain systemic diseases including atherosclerosis and aspiration pneumonia. This organism induces innate responses predominantly through TLR2, which also mediates its ability to induce experimental periodontitis and accelerate atherosclerosis. Using a validated mouse model of intratracheal challenge, we investigated the role of TLR2 in the control of P. gingivalis acute pulmonary infection. TLR2-deficient mice elicited reduc...

Hajishengallis, George; Wang, Min; Bagby, Gregory J.; Nelson, Steve

2008-01-01

130

T-Cell Expression Cloning of Porphyromonas gingivalis Genes Coding for T Helper-Biased Immune Responses during Infection  

OpenAIRE

Exposure of the mouse oral cavity to Porphyromonas gingivalis results in the development of gingivitis and periapical bone loss, which apparently are associated with a Th1 response to bacterial antigens. We have used this infection model in conjunction with direct T-cell expression cloning to identify bacterial antigens that induce a preferential or biased T helper response during the infectious process. A P. gingivalis-specific CD4 T-cell line derived from mice at 3 weeks postchallenge was u...

Gonc?alves, Reginaldo B.; Leshem, Onir; Bernards, Karen; Webb, John R.; Stashenko, Philip P.; Campos-neto, Antonio

2006-01-01

131

Tissue Destruction Induced by Porphyromonas gingivalis Infection in a Mouse Chamber Model Is Associated with Host Tumor Necrosis Factor Generation  

OpenAIRE

Intrachamber challenge with Porphyromonas gingivalis strain 381 in a mouse subcutaneous chamber model results in a local infection that progresses to exfoliation of the chambers within 15 days. This study was designed to elucidate the contribution of host reactions to tissue destruction manifested by chamber exfoliation in animals infected with P. gingivalis. Chamber fluids showed increasing levels of prostaglandin E2 with infection, and the levels of tumor necrosis factor (TNF) in chamber fl...

Lin, Yuh-yih; Huang, Jan-hung; Lai, Yo-yin; Huang, Han-ching; Hu, Suh-woan

2005-01-01

132

Variability of the fimA gene in Porphyromonas gingivalis isolated from periodontitis and non-periodontitis patients  

OpenAIRE

Objective: The goal of this study was to determine the genetic variability of the fimA gene in Porphyromonas gingivalis isolates from Spanish patients. Study Design: Pooled subgingival samples were taken, processed and cultured in non-selective blood agar medium. Pure cultures of one to six isolates per patient were obtained and PCR and PCR-RFLP were used for fimbrillin gene (fimA) type determination of the extracted genomic (DNA). Results: Two hundred and twenty four Porphyromonas gi...

Fabrizi, Simone; Leo?n, Rube?n; Blanc, Vanessa; Herrera, David; Sanz, Mariano

2012-01-01

133

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

Directory of Open Access Journals (Sweden)

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

C.M. Ardila Medina

2010-04-01

134

Honey – a potential agent against Porphyromonas gingivalis: an in vitro study  

Science.gov (United States)

Background Honey has been discussed as a therapeutic option in wound healing since ancient time. It might be also an alternative to the commonly used antimicrobials in periodontitis treatment. The in-vitro study was aimed to determine the antimicrobial efficacy against Porphyromonas gingivalis as a major periodontopathogen. Methods One Manuka and one domestic beekeeper honey have been selected for the study. As a screening, MICs of the honeys against 20 P. gingivalis strains were determined. Contents of methylglyoxal and hydrogen peroxide as the potential antimicrobial compounds were determined. These components (up to 100 mg/l), propolis (up to 200 mg/l) as well as the two honeys (up to 10% w/v) were tested against four P. gingivalis strains in planktonic growth and in a single-species biofilm. Results 2% of Manuka honey inhibited the growth of 50% of the planktonic P. gingivalis, the respective MIC50 of the German beekeeper honey was 5%. Manuka honey contained 1.87 mg/kg hydrogen peroxide and the domestic honey 3.74 mg/kg. The amount of methylglyoxal was found to be 2 mg/kg in the domestic honey and 982 mg/kg in the Manuka honey. MICs for hydrogen peroxide were 10 mg/l - 100 mg/l, for methylglyoxal 5 – 20 mg/l, and for propolis 20 mg/l – 200 mg/l. 10% of both types of honey inhibited the formation of P. gingivalis biofilms and reduced the numbers of viable bacteria within 42 h-old biofilms. Neither a total prevention of biofilm formation nor a complete eradication of a 42 h-old biofilm by any of the tested compounds and the honeys were found. Conclusions Honey acts antibacterial against P. gingivalis. The observed pronounced effects of Manuka honey against planktonic bacteria but not within biofilm can be attributed to methylglyoxal as the characteristic antimicrobial component. PMID:24666777

2014-01-01

135

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

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english Introduction: Periodontitis is a chronic disease that results from an interaction of a mixed bacterial challenge and the host response. Objective: The purposes of this study were to evaluate the IgG serum levels to Porphyromonas gingivalis antigens by ELISA in individuals with different periodontal [...] conditions correlated with clinical parameters, and to analyze the immunoreactivity profiles by Western blotting. Methods: Serum IgG levels against the cell sonicate antigen from P. gingivalis ATCC 33277 of 28 patients with chronic periodontitis (CP), 10 patients with gingivitis (G) and 21 periodontally healthy individuals (H) were measured by ELISA and Western immunoblotting. Results: In the CP group, sera reactivity by ELISA was significantly higher than in the G and H groups (Kruskal-Wallis p

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

2007-06-01

136

Genotype analysis of Porphyromonas gingivalis fimA in Korean adults using new primers.  

Science.gov (United States)

Strains of Porphyromonas gingivalis, a periodontopathic bacterium, are classified into six genotypic variants based on nucleotide sequence differences in the fimA gene encoding FimA. A PCR assay using primer sets specific for each genotype has demonstrated that the most predominant fimA genotype in periodontitis patients is type II, which is now commonly referred to as the periodontitis-associated fimA genotype of P. gingivalis. However, the potential for false type II fimA positives caused by cross-hybridization of type II fimA-specific primers with type Ib fimA has complicated the genotyping. A previous study developed new primers that specifically amplified only the DNA fragment of type II fimA. The aim of the present study was to assess the prevalence of P. gingivalis fimA genotypes in Korean adults and to reconfirm the relationship between type II fimA and periodontitis using the new primers. Among 412 Korean adults, P. gingivalis was detected in 97.5 % of patients and 57.8 % of healthy subjects. Type II fimA was the most widely distributed type among healthy and periodontitis subjects. Organisms with types II, Ib and IV fimA had a significant frequency of occurrence in periodontitis subjects. Statistical analysis, however, revealed that a more significant correlation was found between periodontitis and the occurrence of type Ib fimA. PMID:23264452

Moon, Ji-Hoi; Herr, Yeek; Lee, Hyeon-Woo; Shin, Seung-Il; Kim, Cheul; Amano, Atsuo; Lee, Jin-Yong

2013-09-01

137

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

Science.gov (United States)

Objectives. To examine the interaction between Porphyromonas gingivalis and 3 different orthodontic brackets in vitro, focusing on the effect of an early salivary pellicle and other bacteria on the formation of biofilms. Material and Methods. Mono- and multi-species P. gingivalis biofilms were allowed to form in vitro, on 3 different bracket types (stainless steel, ceramic and plastic) with and without an early salivary pellicle. The brackets were anaerobically incubated for 3 days in Brain Heart Infusion Broth to form biofilms. Bacteria were quantified by trypsin treatment and enumeration of the total viable counts of bacteria recovered. Results. Saliva was found to significantly affect (P biofilm formation of P. gingivalis, with higher numbers for the coated brackets. No significant effect was detected for the impact of the type of biofilm, although on stainless steel and plastic brackets there was a tendency for higher numbers of the pathogen in multi-species biofilms. Bracket material alone was not found to affect the number of bacteria. Conclusions. The salivary pellicle seems to facilitate the adhesion of P. gingivalis and biofilm formation on orthodontic brackets, while the material comprising the brackets does not significantly impact on the number of bacteria. PMID:22315606

Papaioannou, William; Panagopoulos, Athanasios; Koletsi-Kounari, Haroula; Kontou, Efterpi; Makou, Margarita

2012-01-01

138

Histidine Kinase-Mediated Production and Autoassembly of Porphyromonas gingivalis Fimbriae? †  

OpenAIRE

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 enc...

Nishikawa, Kiyoshi; Duncan, Margaret J.

2010-01-01

139

Distribution of Porphyromonas gingivalis Strains with fimA Genotypes in Periodontitis Patients  

OpenAIRE

Fimbriae (FimA) of Porphyromonas gingivalis are filamentous components on the cell surface and are thought to play an important role in the colonization and invasion of periodontal tissues. We previously demonstrated that fimA can be classified into four variants (types I to IV) on the basis of the nucleotide sequences of the fimA gene. In the present study, we attempted to detect the four different fimA genes in saliva and plaque samples isolated from patients with periodontitis using the PC...

Amano, Atsuo; Nakagawa, Ichiro; Kataoka, Kosuke; Morisaki, Ichijiro; Hamada, Shigeyuki

1999-01-01

140

Construction and characterization of a fimA mutant of Porphyromonas gingivalis.  

OpenAIRE

Although fimbriae of Porphyromonas gingivalis have been implicated as playing a major role in adherence to gingival tissue surfaces, no conclusive genetic evidence has yet been obtained. The fimA gene, the determinant for the major fimbrial subunit protein, was cloned and sequenced (D. P. Dickinson, M. A. Kubiniec, F. Yoshimura, and R. J. Genco, J. Bacteriol. 170:1658-1665, 1988). We undertook to inactivate the fimA gene by a homologous recombination technique and examined the fimA mutant for...

Hamada, N.; Watanabe, K.; Sasakawa, C.; Yoshikawa, M.; Yoshimura, F.; Umemoto, T.

1994-01-01

141

Crystallization and preliminary X-ray characterization of prolyl tripeptidyl aminopeptidase from Porphyromonas gingivalis.  

Science.gov (United States)

A recombinant form of prolyl tripeptidyl aminopeptidase from Porphyromonas gingivalis has been crystallized by the hanging-drop vapour-diffusion method using potassium sodium tartrate as a precipitating agent. The crystals belong to the hexagonal space group P6(3)22, with unit-cell parameters a = b = 149.4, c = 159.7 A. The crystals are most likely to contain one subunit of a dimer in the asymmetric unit, with a VM value of 3.14 A3 Da(-1). Diffraction data were collected to 2.1 A resolution using synchrotron radiation at the BL5 station of the Photon Factory. PMID:16511231

Nakajima, Yoshitaka; Ito, Kiyoshi; Xu, Yue; Yamada, Nozomi; Onohara, Yuko; Ito, Takashi; Yoshimoto, Tadashi

2005-12-01

142

fimA Genotypes and Multilocus Sequence Types of Porphyromonas gingivalis from Patients with Periodontitis?  

OpenAIRE

Fimbriae are important virulence factors of pathogenic bacteria, facilitating their attachment to host and bacterial cells. In the periodontal pathogen Porphyromonas gingivalis, the fimA gene is classified into six types (genotypes I, Ib, II, III, IV, and V) on the basis of different nucleotide sequences, with fimA genotypes II and IV being prevalent in isolates from patients with periodontitis. The aims of this study were to examine the distribution of fimA genotypes in a collection of 82 P....

Enersen, Morten; Olsen, Ingar; Kvalheim, Øyvind; Caugant, Dominique A.

2007-01-01

143

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

OpenAIRE

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

144

Localization and function of the accessory protein Mfa3 in Porphyromonas gingivalis Mfa1 fimbriae.  

Science.gov (United States)

The fimbriae of Porphyromonas gingivalis, the causative agent of periodontitis, have been implicated in various aspects of pathogenicity, such as colonization, adhesion and aggregation. Porphyromonas gingivalis ATCC 33277 has two adhesins comprised of the FimA and Mfa1 fimbriae. We characterized the PGN0289 (Mfa3) protein, which is one of the three accessory proteins of Mfa1 fimbriae in P. gingivalis. The Mfa3 protein was present in two different sizes, 40 and 43 kDa, in the cell. The 43-kDa and 40-kDa Mfa3 were detected largely in the inner membrane and the outer membrane, respectively. Purified Mfa1 fimbriae contained the 40-kDa Mfa3 alone. Furthermore, the 40-kDa Mfa3 started with the Ala(44) residue of the deduced amino acid sequence, indicating that the N-terminal region of the nascent protein expressed from the mfa3 gene is processed in the transport step from the inner membrane into fimbriae. Immuno-electron microscopy revealed that Mfa3 localized at the tip of the fimbrial shaft. Interestingly, deletion of the mfa3 gene resulted in the absence of other accessory proteins, PGN0290 and PGN0291, in the purified Mfa1 fimbriae, suggesting that Mfa3 is required for integration of PGN0290 and PGN0291 into fimbriae. A double mutant of mfa3 and fimA genes (phenotype Mfa1 plus, FimA minus) showed increased auto-aggregation and biofilm formation similar to a double mutant of mfa1 and fimA genes (phenotype Mfa1(-) , FimA(-) ). These findings suggest that the tip protein Mfa3 of the Mfa1 fimbriae may function in the integration of accessory proteins and in the colonization of P. gingivalis. PMID:24118823

Hasegawa, Y; Nagano, K; Ikai, R; Izumigawa, M; Yoshida, Y; Kitai, N; Lamont, R J; Murakami, Y; Yoshimura, F

2013-12-01

145

Role of Differential Expression of Streptococcal Arginine Deiminase in Inhibition of fimA Expression in Porphyromonas gingivalis?  

OpenAIRE

Streptococcus cristatus ArcA inhibits production of a major adhesin, FimA, in Porphyromonas gingivalis, a primary periodontal pathogen. In this study, we demonstrate the differential expression of arcA in two streptococcal species. The expression level of arcA in streptococci appears to be controlled by both cis and trans elements.

Lin, Xinghua; Lamont, Richard J.; Wu, Jie; Xie, Hua

2008-01-01

146

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

147

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

148

Dental implant surface treatments may modulate cytokine secretion in Porphyromonas gingivalis-stimulated human gingival fibroblasts: a comparative study.  

Science.gov (United States)

Peri-implantitis is an inflammation that affects dental implants and can lead to implant loss. The aim of this study was to analyze the in vitro effect of different implant surface treatments on cytokine production by human gingival fibroblasts (HGFs) stimulated or not with Porphyromonas gingivalis lipopolysaccharide (PgLPS). Six different titanium implants were tested: turned, sandblasted, anodized, acid-etched, TiO2-blasted/acid-etched, and grit-blasted/acid-etched. HGFs were seeded with each implant in a 6-well plate and assayed before LPS treatment (-LPS) or after 36 h of LPS (+LPS) treatment. Protein concentrations were measured using a Pierce bicinchoninic acid (BCA) assay and cytokine secretions were analyzed using a multiplex cytokine array. Scanning electron microscopy was performed for sterile implants and after cell attachment. Protein levels were consistent across all implants indicating that cell growth was uniform (p?>?0.05). Sandblasted and turned surfaces significantly increased the secretion of interleukin (IL)-6, -8, -10, MCP-1 and VEGF (p?

Stavroullakis, Alexander; Brito, Carlos; Chen, Hong Yang; Bajenova, Elena; Prakki, Anuradha; Nogueira-Filho, Getulio

2015-03-01

149

Porphyromonas gingivalis oral infection exacerbates the development and severity of collagen-induced arthritis  

Science.gov (United States)

Introduction Clinical studies suggest a direct influence of periodontal disease (PD) on serum inflammatory markers and disease assessment of patients with established rheumatoid arthritis (RA). However, the influence of PD on arthritis development remains unclear. This investigation was undertaken to determine the contribution of chronic PD to immune activation and development of joint inflammation using the collagen-induced arthritis (CIA) model. Methods DBA1/J mice orally infected with Porphyromonas gingivalis were administered with collagen II (CII) emulsified in complete Freund’s adjuvant (CFA) or incomplete Freund’s adjuvant (IFA) to induce arthritis. Arthritis development was assessed by visual scoring of paw swelling, caliper measurement of the paws, mRNA expression, paw micro-computed tomography (micro-CT) analysis, histology, and tartrate resistant acid phosphatase for osteoclast detection (TRAP)-positive immunohistochemistry. Serum and reactivated splenocytes were evaluated for cytokine expression. Results Mice induced for PD and/or arthritis developed periodontal disease, shown by decreased alveolar bone and alteration of mRNA expression in gingival tissues and submandibular lymph nodes compared to vehicle. P. gingivalis oral infection increased paw swelling and osteoclast numbers in mice immunized with CFA/CII. Arthritis incidence and severity were increased by P. gingivalis in mice that received IFA/CII immunizations. Increased synovitis, bone erosions, and osteoclast numbers in the paws were observed following IFA/CII immunizations in mice infected with P gingivalis. Furthermore, cytokine analysis showed a trend toward increased serum Th17/Th1 ratios when P. gingivalis infection was present in mice receiving either CFA/CII or IFA/CII immunizations. Significant cytokine increases induced by P. gingivalis oral infection were mostly associated to Th17-related cytokines of reactivated splenic cells, including IL-1?, IL-6, and IL-22 in the CFA/CII group and IL-1?, tumor necrosis factor-?, transforming growth factor-?, IL-6 and IL-23 in the IFA/CII group. Conclusions Chronic P. gingivalis oral infection prior to arthritis induction increases the immune system activation favoring Th17 cell responses, and ultimately accelerating arthritis development. These results suggest that chronic oral infection may influence RA development mainly through activation of Th17-related pathways. PMID:24456966

2013-01-01

150

High In Vitro Antibacterial Activity of Pac-525 against Porphyromonas gingivalis Biofilms Cultured on Titanium.  

Science.gov (United States)

In order to investigate the potential of short antimicrobial peptides (AMPs) as alternative antibacterial agents during the treatment of peri-implantitis, the cytotoxic activity of three short AMPs, that is, Pac-525, KSL-W, and KSL, was determined using the MTT assay. The antimicrobial activity of these AMPs, ranging in concentration from 0.0039?mg/mL to 0.5?mg/mL, against the predominant planktonic pathogens, including Streptococcus sanguis, Fusobacterium nucleatum, and Porphyromonas gingivalis, involved in peri-implantitis was investigated. Furthermore, 2-day-old P. gingivalis biofilms cultured on titanium surfaces were treated with Pac-525 and subsequently observed and analysed using confocal laser scanning microscopy (CLSM). The average cell proliferation curve indicated that there was no cytotoxicity due to the three short AMPs. The minimum inhibitory concentration and minimum bactericidal concentration values of Pac-525 were 0.0625?mg/mL and 0.125?mg/mL, respectively, for P. gingivalis and 0.0078?mg/mL and 0.0156?mg/mL, respectively, for F. nucleatum. Using CLSM, we confirmed that compared to 0.1% chlorhexidine, 0.5?mg/mL of Pac-525 caused a significant decrease in biofilm thickness and a decline in the percentage of live bacteria. These data indicate that Pac-525 has unique properties that might make it suitable for the inhibition the growth of pathogenic bacteria around dental implants. PMID:25710035

Li, Ji-Yin; Wang, Xue-Jin; Wang, Li-Na; Ying, Xiao-Xia; Ren, Xiang; Liu, Hui-Ying; Xu, Li; Ma, Guo-Wu

2015-01-01

151

VimA – dependent modulation of the secretome in Porphyromonas gingivalis  

Science.gov (United States)

The VimA protein of Porphyromonas gingivalis is a multifunctional protein involved in cell surface biogenesis. To further determine if its acetyl coenzyme A (acetyl-CoA) transfer and putative sorting functions can affect the secretome, its role in peptidoglycan biogenesis and effects on the extracellular proteins of P. gingivalis FLL92, a vimA-defective mutant, were evaluated. There were structural and compositional differences in the peptidoglycan of P. gingivalis FLL92 compared to the wild-type strain. Sixty-eight proteins were present only in the extracellular fraction of FLL92. Fifteen proteins present in the extracellular fraction of the parent strain were missing in the vimA-defective mutant. These proteins had protein sorting characteristics which included a C terminal motif with a common consensus Gly-Gly – Cterm pattern and polar tail consisting of aromatic amino acid residues. These observations suggest that the VimA protein is likely involved in peptidoglycan synthesis, and corroborates our previous report, which suggests a role in protein sorting. PMID:23134608

Osbourne, D.; Aruni, A.Wilson; Dou, Y.; Perry, C.; Boskovic, D.S.; Roy, F.; Fletcher, H. M.

2012-01-01

152

VimA-dependent modulation of the secretome in Porphyromonas gingivalis.  

Science.gov (United States)

The VimA protein of Porphyromonas gingivalis is a multifunctional protein involved in cell surface biogenesis. To further determine if its acetyl coenzyme A (acetyl-CoA) transfer and putative sorting functions can affect the secretome, its role in peptidoglycan biogenesis and effects on the extracellular proteins of P. gingivalis FLL92, a vimA-defective mutant, were evaluated. There were structural and compositional differences in the peptidoglycan of P. gingivalis FLL92 compared with the wild-type strain. Sixty-eight proteins were present only in the extracellular fraction of FLL92. Fifteen proteins present in the extracellular fraction of the parent strain were missing in the vimA-defective mutant. These proteins had protein sorting characteristics that included a C-terminal motif with a common consensus Gly-Gly-CTERM pattern and a polar tail consisting of aromatic amino acid residues. These observations suggest that the VimA protein is likely involved in peptidoglycan synthesis, and corroborates our previous report, which suggests a role in protein sorting. PMID:23134608

Osbourne, D; Aruni, A Wilson; Dou, Y; Perry, C; Boskovic, D S; Roy, F; Fletcher, H M

2012-12-01

153

VimA mediates multiple functions that control virulence in Porphyromonas gingivalis.  

Science.gov (United States)

Porphyromonas gingivalis, a black-pigmented, gram-negative anaerobe, is an important etiological agent of periodontal disease. Its ability to survive in the periodontal pocket and orchestrate the microbial/host activities that can lead to disease suggest that P. gingivalis possesses a complex regulatory network involving transcriptional and post-transcriptional mechanisms. The vimA (virulence modulating) gene is part of the 6.15-kb bcp-recA-vimA-vimE-vimF-aroG locus and plays a role in oxidative stress resistance. In addition to the glycosylation and anchorage of several surface proteins including the gingipains, VimA can also modulate sialylation, acetyl coenzyme A transfer, lipid A and its associated proteins and may be involved in protein sorting and transport. In this review, we examine the multifunctional role of VimA and discuss its possible involvement in a major regulatory network important for survival and virulence regulation in P. gingivalis. It is postulated that the multifunction of VimA is modulated via a post-translational mechanism involving acetylation. PMID:23279905

Aruni, A W; Robles, A; Fletcher, H M

2013-06-01

154

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

Science.gov (United States)

Recombinant VimA protein can interact with the gingipains and several other proteins that may play a role in its biogenesis in Porphyromonas gingivalis. In silico analysis of PG2096, a hypothetical protein that was shown to interact with VimA, suggests that it may have environmental stress resistance properties. To further evaluate the role(s) of PG2096, the predicted open reading frame was PCR amplified from P. gingivalis W83 and insertionally inactivated using the ermF-ermAM antibiotic-resistance cassette. One randomly chosen PG2096-defective mutant created by allelic exchange and designated FLL205 was further characterized. Under normal growth conditions at 37 °C, Arg-X and Lys-X gingipain activities in FLL205 were reduced by approximately 35?% and 21?%, respectively, compared to the wild-type strain. However, during prolonged growth at an elevated temperature of 42 °C, Arg-X activity was increased by more than 40?% in FLL205 in comparison to the wild-type strain. In addition, the PG2096-defective mutant was more resistant to oxidative stress when treated with 0.25 mM hydrogen peroxide. Taken together these results suggest that the PG2096 gene, designated regT (regulator of gingipain activity at elevated temperatures), may be involved in regulating gingipain activity at elevated temperatures and be important in oxidative stress resistance in P. gingivalis. PMID:20595264

Vanterpool, E; Aruni, A Wilson; Roy, F; Fletcher, H M

2010-10-01

155

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

Directory of Open Access Journals (Sweden)

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

156

Porphyromonas gingivalis-dendritic cell interactions: consequences for coronary artery disease.  

Science.gov (United States)

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. PMID:21523219

Zeituni, Amir E; Carrion, Julio; Cutler, Christopher W

2010-01-01

157

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 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

158

The capsule of Porphyromonas gingivalis reduces the immune response of human gingival fibroblasts  

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Full Text Available Abstract Background Periodontitis is a bacterial infection of the periodontal tissues. The Gram-negative anaerobic bacterium Porphyromonas gingivalis is considered a major causative agent. One of the virulence factors of P. gingivalis is capsular polysaccharide (CPS. Non-encapsulated strains have been shown to be less virulent in mouse models than encapsulated strains. Results To examine the role of the CPS in host-pathogen interactions we constructed an insertional isogenic P. gingivalis knockout in the epimerase-coding gene epsC that is located at the end of the CPS biosynthesis locus. This mutant was subsequently shown to be non-encapsulated. K1 capsule biosynthesis could be restored by in trans expression of an intact epsC gene. We used the epsC mutant, the W83 wild type strain and the complemented mutant to challenge human gingival fibroblasts to examine the immune response by quantification of IL-1?, IL-6 and IL-8 transcription levels. For each of the cytokines significantly higher expression levels were found when fibroblasts were challenged with the epsC mutant compared to those challenged with the W83 wild type, ranging from two times higher for IL-1? to five times higher for IL-8. Conclusions These experiments provide the first evidence that P. gingivalis CPS acts as an interface between the pathogen and the host that may reduce the host's pro-inflammatory immune response. The higher virulence of encapsulated strains may be caused by this phenomenon which enables the bacteria to evade the immune system.

van Winkelhoff Arie J

2010-01-01

159

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

Science.gov (United States)

The prevention and treatment of periodontitis requires not only the control of causative pathogens, especially Porphyromonas gingivalis, but also the regulation of inflammatory immune response. Investigating auxiliary drugs for periodontitis during conventional treatments is, thus, quite important. Capsaicin, an agonist for the vanilloid receptor subtype 1 (TRPV1), due to its bacteriostatic activity against Gram-negative bacteria and anti-inflammatory effects, appears to be a promising drug. In this work, the antimicrobial activity of capsaicin against P. gingivalis and biofilm formation, inflammatory cytokine levels in experimental periodontitis, osteoclast precursor proliferation, and osteoclastogenesis in vitro were fully investigated. The results showed that capsaicin inhibited P. gingivalis growth with a minimum inhibitory concentration (MIC) and a minimum bactericidal concentration (MBC) of 16 and 64 mg/l, respectively. Capsaicin also inhibited P. gingivalis biofilm formation, with minimum biofilm inhibition concentrations MBIC50 and MBIC90 of 16 and 32 mg/l, respectively, and reduced pre-formed biofilms' viability with a minimum biofilm reduction concentration MBRC50 of 64 mg/l, as demonstrated by confocal laser scanning microscopy. In experimental periodontitis, except for IL-10, TNF-?, IL-1?, IL-6, IL-12, and iNOS were depressed after capsaicin treatment. Moreover, capsaicin also suppressed osteoclast precursor proliferation and osteoclastogenesis, as demonstrated by NF-?B p65. However, this favorable effect was attenuated by the TRPV1 antagonist, camphor. It, thus, suggests that capsaicin is a potential drug for the auxiliary treatment of periodontitis. TRPV1 activation may involve in beneficial roles of capsaicin on periodontitis. PMID:23955115

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

2014-02-01

160

Hemoglobinase activity of the lysine gingipain protease (Kgp) of Porphyromonas gingivalis W83.  

Science.gov (United States)

Porphyromonas gingivalis, an important periodontal disease pathogen, forms black-pigmented colonies on blood agar. Pigmentation is believed to result from accumulation of iron protoporphyrin IX (FePPIX) derived from erythrocytic hemoglobin. The Lys-X (Lys-gingipain) and Arg-X (Arg-gingipain) cysteine proteases of P. gingivalis bind and degrade erythrocytes. We have observed that mutations abolishing activity of the Lys-X-specific cysteine protease, Kgp, resulted in loss of black pigmentation of P. gingivalis W83. Because the hemagglutinating and hemolytic potentials of mutant strains were reduced but not eliminated, we hypothesized that this protease played a role in acquisition of FePPIX from hemoglobin. In contrast to Arg-gingipain, Lys-gingipain was not inhibited by hemin, suggesting that this protease played a role near the cell surface where high concentrations of hemin confer the black pigmentation. Human hemoglobin contains 11 Lys residues in the alpha chain and 10 Lys residues in the beta chain. In contrast, there are only three Arg residues in each of the alpha and beta chains. These observations are consistent with human hemoglobin being a preferred substrate for Lys-gingipain but not Arg-gingipain. The ability of the Lys-gingipain to cleave human hemoglobin at Lys residues was confirmed by electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry of hemoglobin fragments resulting from digestion with the purified protease. We were able to detect several of the predicted hemoglobin fragments rendered by digestion with purified Lys-gingipain. Thus, we postulate that the Lys-gingipain of P. gingivalis is a hemoglobinase which plays a role in heme and iron uptake by effecting the accumulation of FePPIX on the bacterial cell surface. PMID:10438761

Lewis, J P; Dawson, J A; Hannis, J C; Muddiman, D; Macrina, F L

1999-08-01

161

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

Science.gov (United States)

The effects of sugar alcohols such as erythritol, xylitol, and sorbitol on periodontopathic biofilm are poorly understood, though they have often been reported to be non-cariogenic sweeteners. In the present study, we evaluated the efficacy of sugar alcohols for inhibiting periodontopathic biofilm formation using a heterotypic biofilm model composed of an oral inhabitant Streptococcus gordonii and a periodontal pathogen Porphyromonas gingivalis. Confocal microscopic observations showed that the most effective reagent to reduce P. gingivalis accumulation onto an S. gordonii substratum was erythritol, as compared with xylitol and sorbitol. In addition, erythritol moderately suppressed S. gordonii monotypic biofilm formation. To examine the inhibitory effects of erythritol, we analyzed the metabolomic profiles of erythritol-treated P. gingivalis and S. gordonii cells. Metabolome analyses using capillary electrophoresis time-of-flight mass spectrometry revealed that a number of nucleic intermediates and constituents of the extracellular matrix, such as nucleotide sugars, were decreased by erythritol in a dose-dependent manner. Next, comparative analyses of metabolites of erythritol- and sorbitol-treated cells were performed using both organisms to determine the erythritol-specific effects. In P. gingivalis, all detected dipeptides, including Glu-Glu, Ser-Glu, Tyr-Glu, Ala-Ala and Thr-Asp, were significantly decreased by erythritol, whereas they tended to be increased by sorbitol. Meanwhile, sorbitol promoted trehalose 6-phosphate accumulation in S. gordonii cells. These results suggest that erythritol has inhibitory effects on dual species biofilm development via several pathways, including suppression of growth resulting from DNA and RNA depletion, attenuated extracellular matrix production, and alterations of dipeptide acquisition and amino acid metabolism. PMID:23890177

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

2013-12-01

162

Porphyromonas gingivalis Gingipains Selectively Reduce CD14 Expression, Leading to Macrophage Hyporesponsiveness to Bacterial Infection.  

Science.gov (United States)

Cysteine proteases (gingipains) from Porphyromonas gingivalis are key virulence factors in chronic periodontitis. Innate immune receptors CD14, Toll-like receptor (TLR) 2 and TLR4 are important in P. gingivalis recognition. We examined the ability of gingipains to cleave CD14, TLR2 and TLR4, and the consequences for the cellular response to bacterial challenge. Macrophages were exposed to Arg (RgpA and RgpB)- and Lys (Kgp)-gingipains, and residual expression of TLR2, TLR4 and CD14 was determined by flow cytometry. The cellular response to live bacteria following exposure to purified gingipains was evaluated by TNF? production and bacterial phagocytosis. RgpA and Kgp decreased CD14 detection in a concentration (p = 0.0000002)- and time (p = 0.03)-dependent manner, whereas RgpB had no significant effect. TLR2 and TLR4 expression were unaffected. Reduction in CD14 expression was more efficient with Lys-gingipain than with Arg-gingipain. A reduced CD14 surface level correlated with decreased TNF? secretion and bacterial phagocytosis following challenge with live P. gingivalis, but the response to heat-killed bacteria was unaffected. Therefore, gingipains reduce CD14 expression without affecting expression of the bacterial-sensing TLRs. Reduced CD14 expression depends on the gingipain hemagglutinin/adhesion site and results in macrophage hyporesponsiveness to bacterial challenge. Further studies are needed to determine if reduced CD14 expression is linked to periodontitis induced by P. gingivalis. © 2014 S. Karger AG, Basel. PMID:25228314

Wilensky, Asaf; Tzach-Nahman, Rinat; Potempa, Jan; Shapira, Lior; Nussbaum, Gabriel

2015-01-01

163

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

164

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

Science.gov (United States)

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

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

2001-01-01

165

In situ visualization of plasma cells producing antibodies reactive to Porphyromonas gingivalis in periodontitis: the application of the enzyme-labeled antigen method  

OpenAIRE

Porphyromonas gingivalis is a keystone periodontal pathogen. Histologocally, the gingival tissue in periodontitis shows dense infiltration of plasma cells. However, antigens recognized by antibodies secreted from the immunocytes remain unknown. The enzyme-labeled antigen method was applied to detecting plasma cells producing P. gingivalis-specific antibodies in biopsied gingival tissue of periodontitis. N-terminally biotinylated P. gingivalis antigens, Ag53 and four gingipain domains (Arg-p...

Mizutani, Y.; Tsuge, S.; Takeda, H.; Hasegawa, Y.; Shiogama, K.; Onouchi, T.; Inada, K.; Sawasaki, T.; Tsutsumi, Y.

2014-01-01

166

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

OpenAIRE

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 a...

Wu, Jie; Lin, Xinghua; Xie, Hua

2007-01-01

167

Anchoring and length regulation of Porphyromonas gingivalis Mfa1 fimbriae by the downstream gene product Mfa2  

OpenAIRE

Porphyromonas gingivalis, a causative agent of periodontitis, has at least two types of thin, single-stranded fimbriae, termed FimA and Mfa1 (according to the names of major subunits), which can be discriminated by filament length and by the size of their major fimbrilin subunits. FimA fimbriae are long filaments that are easily detached from cells, whereas Mfa1 fimbriae are short filaments that are tightly bound to cells. However, a P. gingivalis ATCC 33277-derived mutant deficient in mfa2, ...

Hasegawa, Yoshiaki; Iwami, Jun; Sato, Keiko; Park, Yoonsuk; Nishikawa, Kiyoshi; Atsumi, Tatsuo; Moriguchi, Keiichi; Murakami, Yukitaka; Lamont, Richard J.; Nakamura, Hiroshi; Ohno, Norikazu; Yoshimura, Fuminobu

2009-01-01

168

OxyR is involved in coordinate regulation of expression of fimA and sod genes in Porphyromonas gingivalis  

OpenAIRE

Survival of Porphyromonas gingivalis in the constantly changing oral environment depends on its ability to alter gene expression. We demonstrate here that P. gingivalis activates superoxide dismutase expression in response to oxidative stress and represses expression of FimA, a subunit of major fimbriae. Coordinated expression of fimA and sod is regulated by the redox-sensing transcription factor OxyR. Mutations in the oxyR gene result in a decreased expression of sod and in an elevated expre...

Wu, Jie; Lin, Xinghua; Xie, Hua; Burne, Robert

2008-01-01

169

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

OpenAIRE

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

170

Genetic and antigenic analyses of Porphyromonas gingivalis FimA fimbriae.  

Science.gov (United States)

The periodontal pathogen Porphyromonas gingivalis generally expresses two distinct fimbriae, FimA and Mfa1, which play a role in biofilm formation. The fimA gene that encodes FimA fimbrilin is polymorphic, and polymerase chain reaction analysis has identified six genotypes called types I-V and Ib. We found recently that fimbriae exhibit antigenic heterogeneity among the genotypes. In the present study, we analysed the fimA DNA sequences of 84 strains of P. gingivalis and characterized the antigenicity of FimA fimbriae. Strains analysed here comprised 10, 16, 29, 13, 10 and 6 strains of types I, Ib, II, III, IV and V, respectively. DNA sequencing revealed that type Ib does not represent a single cluster and that type II sequences are remarkably diverse. In contrast, the fimA sequences of the other types were relatively homogeneous. Antigenicity was investigated using antisera elicited by pure FimA fimbriae of types I-V. Antigenicity correlated generally with the respective genotype. Type Ib strains were recognized by type I antisera. However, some strains showed cross-reactivity, especially, many type II strains reacted with type III antisera. The levels of fimbrial expression were highly variable, and expression was positively correlated with ability of biofilm formation on a saliva-coated plate. Further, two strains without FimA and Mfa1 fimbriae expressed fimbrial structures, suggesting that the strains produce other types of fimbriae. PMID:23809984

Nagano, K; Abiko, Y; Yoshida, Y; Yoshimura, F

2013-10-01

171

Pyrano-isoflavans from Glycyrrhiza uralensis with antibacterial activity against Streptococcus mutans and Porphyromonas gingivalis.  

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Continuing investigation of fractions from a supercritical fluid extract of Chinese licorice (Glycyrrhiza uralensis) roots has led to the isolation of 12 phenolic compounds, of which seven were described previously from this extract. In addition to these seven metabolites, four known components, 1-methoxyerythrabyssin II (4), 6,8-diprenylgenistein, gancaonin G (5), and isoglycyrol (6), and one new isoflavan, licorisoflavan C (7), were characterized from this material for the first time. Treatment of licoricidin (1) with palladium chloride afforded larger amounts of 7 and also yielded two new isoflavans, licorisoflavan D (8), which was subsequently detected in the licorice extract, and licorisoflavan E (9). Compounds 1-9 were evaluated for their antibacterial activities against the cariogenic Streptococcus mutans and the periodontopathogenic Porphyromonas gingivalis. Licoricidin (1), licorisoflavan A (2), and 7-9 showed antibacterial activity against P. gingivalis (MICs of 1.56-12.5 ?g/mL). The most potent activity against S. mutans was obtained with 7 (MIC of 6.25 ?g/mL), followed by 1 and 9 (MIC of 12.5 ?g/mL). This study provides further evidence for the therapeutic potential of licorice extracts for the treatment and prevention of oral infections. PMID:24479468

Villinski, Jacquelyn R; Bergeron, Chantal; Cannistra, Joseph C; Gloer, James B; Coleman, Christina M; Ferreira, Daneel; Azelmat, Jabrane; Grenier, Daniel; Gafner, Stefan

2014-03-28

172

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

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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

173

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

DEFF Research Database (Denmark)

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

174

Distinct roles of long/short fimbriae and gingipains in homotypic biofilm development by Porphyromonas gingivalis  

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Full Text Available Abstract Background Porphyromonas gingivalis, a periodontal pathogen, expresses a number of virulence factors, including long (FimA and short (Mfa fimbriae as well as gingipains comprised of arginine-specific (Rgp and lysine-specific (Kgp cysteine proteinases. The aim of this study was to examine the roles of these components in homotypic biofilm development by P. gingivalis, as well as in accumulation of exopolysaccharide in biofilms. Results Biofilms were formed on saliva-coated glass surfaces in PBS or diluted trypticase soy broth (dTSB. Microscopic observation showed that the wild type strain formed biofilms with a dense basal monolayer and dispersed microcolonies in both PBS and dTSB. A FimA deficient mutant formed patchy and small microcolonies in PBS, but the organisms proliferated and formed a cohesive biofilm with dense exopolysaccharides in dTSB. A Mfa mutant developed tall and large microcolonies in PBS as well as dTSB. A Kgp mutant formed markedly thick biofilms filled with large clumped colonies under both conditions. A RgpA/B double mutant developed channel-like biofilms with fibrillar and tall microcolonies in PBS. When this mutant was studied in dTSB, there was an increase in the number of peaks and the morphology changed to taller and loosely packed biofilms. In addition, deletion of FimA reduced the autoaggregation efficiency, whereas autoaggregation was significantly increased in the Kgp and Mfa mutants, with a clear association with alteration of biofilm structures under the non-proliferation condition. In contrast, this association was not observed in the Rgp-null mutants. Conclusion These results suggested that the FimA fimbriae promote initial biofilm formation but exert a restraining regulation on biofilm maturation, whereas Mfa and Kgp have suppressive and regulatory roles during biofilm development. Rgp controlled microcolony morphology and biovolume. Collectively, these molecules seem to act coordinately to regulate the development of mature P. gingivalis biofilms.

Tribble Gena D

2009-05-01

175

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 background. In contrast to the wild type, gingipain activity was increased in P. gingivalis FLL406, a vimA chimeric strain. P. gingivalis FLL451 had a five times higher biofilm-forming capacity than the parent strain. HeLa cells incubated with P. gingivalis FLL92 showed a decrease in invasion, in contrast to P. gingivalis FLL451 and FLL406, which showed increases of 30 and 40%, respectively. VimA mediated coenzyme A (CoA) transfer to isoleucine and reduced branched-chain amino acid metabolism. The lipid A content and associated proteins were altered in the vimA-defective mutants. The VimA chimera interacted with several proteins which were found to have an LXXTG motif, similar to the sorting motif of gram-positive organisms. All the proteins had an N-terminal signal sequence with a putative sorting signal of L(P/T/S)X(T/N/D)G and two unique signatures of EXGXTX and HISXXGXG, in addition to a polar tail. Taken together, these observations further confirm the multifunctional role of VimA in modulating virulence possibly through its involvement in acetyl-CoA transfer and lipid A synthesis and possibly by protein sorting. PMID:22144476

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

2012-02-01

176

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

OpenAIRE

Objectives: The aim of this study was to determine the prevalence of the different fimA genotypes of Porphyromonas gingivalis in adult Spanish patients with chronic periodontitis, patients with gingivitis and periodontally healthy subjects, and the relationship between these genotypes and other periodontopathogenic bacteria. Study design: Samples of subgingival plaque were taken from 86 patients (33 with chronic periodontitis, 16 with gingivitis, and 37 periodontally healthy) in the course...

Puig-silla, Miriam; Dasi?-ferna?nde, Francisco; Montiel-company, Jose?-mari?a; Almerich-silla, Jose?-manuel

2012-01-01

177

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

DEFF Research Database (Denmark)

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

178

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

OpenAIRE

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 we...

Mrudula Patel; Roxanne Naidoo; Foluso John Owotade

2013-01-01

179

Oral mucosal lipids are antibacterial against Porphyromonas gingivalis, induce ultrastructural damage, and alter bacterial lipid and protein compositions  

OpenAIRE

Oral mucosal and salivary lipids exhibit potent antimicrobial activity for a variety of Gram-positive and Gram-negative bacteria; however, little is known about their spectrum of antimicrobial activity or mechanisms of action against oral bacteria. In this study, we examine the activity of two fatty acids and three sphingoid bases against Porphyromonas gingivalis, an important colonizer of the oral cavity implicated in periodontitis. Minimal inhibitory concentrations, minimal bactericidal con...

Fischer, Carol L.; Walters, Katherine S.; Drake, David R.; Dawson, Deborah V.; Blanchette, Derek R.; Brogden, Kim A.; Wertz, Philip W.

2013-01-01

180

Growth of Porphyromonas gingivalis, Treponema denticola, T. pectinovorum, T. socranskii, and T. vincentii in a chemically defined medium.  

OpenAIRE

A chemically defined medium, OMIZ (Oral Microbiology and Immunology, Zürich)-W1 was developed. Medium OMIZ-W1 supports the long-term proliferation of a wide range of oral anaerobes, including representative strains of four Treponema species and Porphyromonas gingivalis. High concentrations of ascorbic acid and ammonium ions proved to be important for the growth of these organisms. T. denticola CD-1 grew in the absence of polyamines and long-chain fatty acids, T. pectinovorum and T. socranski...

Wyss, C.

1992-01-01

181

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

OpenAIRE

This study assessed the effect of periodontal therapy on specific serum antibody concentration, expressed as titer, and antibody binding strength, expressed as relative avidity. The immune responses to Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans were investigated. Antibody titer was assayed by enzyme-linked immunosorbent assay (ELISA) and relative avidity was measured by thiocyanate elution in 17 adult periodontitis patients before and after therapy. Immunoglobulin G (Ig...

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

1995-01-01

182

The profile of Porphyromonas gingivalis kgp biotype and fimA genotype mosaic in subgingival plaque samples.  

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Combined analysis of allelic variation of the virulence-associated, strain-specific lys-gingipain gene (kgp) and major fimbrial gene (fimA) of Porphyromonas gingivalis was undertaken in 116 subgingival plaque samples to understand the kgp biotype and fimA genotype profile in a subject-specific manner. Allelic variation in the polyadhesin domain of kgp from P. gingivalis strains 381 (ATCC 33277), HG66 and W83 generated four isoforms corresponding to four biotypes of P. gingivalis. Similarly, variation in the fimA subunit of the fimA gene cluster of P. gingivalis resulted in six fimA genotypes. Strain-specific differential PCR was performed for kgp and fimA using DNA isolated from subgingival plaque samples. Our findings demonstrate that all of the P. gingivalis kgp biotypes detected in this study were predominantly associated with the fimA II genotype. Dominance of kgp biotypes 381 or HG66 combined with fimA II fimbriae could imply an adaptive strategy by P. gingivalis to generate the fittest strains for survival in the host environment. PMID:25353706

Nadkarni, Mangala A; Chhour, Kim-Ly; Chapple, Cheryl C; Nguyen, Ky-Anh; Hunter, Neil

2014-12-01

183

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

184

Porphyromonas gingivalis Evasion of Autophagy and Intracellular Killing by Human Myeloid Dendritic Cells Involves DC-SIGN-TLR2 Crosstalk  

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Signaling via pattern recognition receptors (PRRs) expressed on professional antigen presenting cells, such as dendritic cells (DCs), is crucial to the fate of engulfed microbes. Among the many PRRs expressed by DCs are Toll-like receptors (TLRs) and C-type lectins such as DC-SIGN. DC-SIGN is targeted by several major human pathogens for immune-evasion, although its role in intracellular routing of pathogens to autophagosomes is poorly understood. Here we examined the role of DC-SIGN and TLRs in evasion of autophagy and survival of Porphyromonas gingivalis in human monocyte-derived DCs (MoDCs). We employed a panel of P. gingivalis isogenic fimbriae deficient strains with defined defects in Mfa-1 fimbriae, a DC-SIGN ligand, and FimA fimbriae, a TLR2 agonist. Our results show that DC-SIGN dependent uptake of Mfa1+P. gingivalis strains by MoDCs resulted in lower intracellular killing and higher intracellular content of P. gingivalis. Moreover, Mfa1+P. gingivalis was mostly contained within single membrane vesicles, where it survived intracellularly. Survival was decreased by activation of TLR2 and/or autophagy. Mfa1+P. gingivalis strain did not induce significant levels of Rab5, LC3-II, and LAMP1. In contrast, P. gingivalis uptake through a DC-SIGN independent manner was associated with early endosomal routing through Rab5, increased LC3-II and LAMP-1, as well as the formation of double membrane intracellular phagophores, a characteristic feature of autophagy. These results suggest that selective engagement of DC-SIGN by Mfa-1+P. gingivalis promotes evasion of antibacterial autophagy and lysosome fusion, resulting in intracellular persistence in myeloid DCs; however TLR2 activation can overcome autophagy evasion and pathogen persistence in DCs. PMID:25679217

El-Awady, Ahmed R.; Miles, Brodie; Scisci, Elizabeth; Kurago, Zoya B.; Palani, Chithra D.; Arce, Roger M.; Waller, Jennifer L.; Genco, Caroline A.; Slocum, Connie; Manning, Matthew; Schoenlein, Patricia V.; Cutler, Christopher W.

2015-01-01

185

Porphyromonas gingivalis Evasion of Autophagy and Intracellular Killing by Human Myeloid Dendritic Cells Involves DC-SIGN-TLR2 Crosstalk.  

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Signaling via pattern recognition receptors (PRRs) expressed on professional antigen presenting cells, such as dendritic cells (DCs), is crucial to the fate of engulfed microbes. Among the many PRRs expressed by DCs are Toll-like receptors (TLRs) and C-type lectins such as DC-SIGN. DC-SIGN is targeted by several major human pathogens for immune-evasion, although its role in intracellular routing of pathogens to autophagosomes is poorly understood. Here we examined the role of DC-SIGN and TLRs in evasion of autophagy and survival of Porphyromonas gingivalis in human monocyte-derived DCs (MoDCs). We employed a panel of P. gingivalis isogenic fimbriae deficient strains with defined defects in Mfa-1 fimbriae, a DC-SIGN ligand, and FimA fimbriae, a TLR2 agonist. Our results show that DC-SIGN dependent uptake of Mfa1+P. gingivalis strains by MoDCs resulted in lower intracellular killing and higher intracellular content of P. gingivalis. Moreover, Mfa1+P. gingivalis was mostly contained within single membrane vesicles, where it survived intracellularly. Survival was decreased by activation of TLR2 and/or autophagy. Mfa1+P. gingivalis strain did not induce significant levels of Rab5, LC3-II, and LAMP1. In contrast, P. gingivalis uptake through a DC-SIGN independent manner was associated with early endosomal routing through Rab5, increased LC3-II and LAMP-1, as well as the formation of double membrane intracellular phagophores, a characteristic feature of autophagy. These results suggest that selective engagement of DC-SIGN by Mfa-1+P. gingivalis promotes evasion of antibacterial autophagy and lysosome fusion, resulting in intracellular persistence in myeloid DCs; however TLR2 activation can overcome autophagy evasion and pathogen persistence in DCs. PMID:25679217

El-Awady, Ahmed R; Miles, Brodie; Scisci, Elizabeth; Kurago, Zoya B; Palani, Chithra D; Arce, Roger M; Waller, Jennifer L; Genco, Caroline A; Slocum, Connie; Manning, Matthew; Schoenlein, Patricia V; Cutler, Christopher W

2015-02-01

186

[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

187

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

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

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

2013-01-01

188

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

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

YANTI

2010-03-01

189

Subcutaneous vaccination with Porphyromonas gingivalis ameliorates periodontitis by modulating Th17/Treg imbalance in a murine model.  

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To date, Porphyromonas gingivalis (P. gingivalis) vaccination has been studied only in animals, and no effective prophylactic human periodontal vaccine has been developed, with the reason for the failure of prophylactic human periodontal vaccines unknown. T helper 17 cell (Th17)/regulatory T (Treg) cell responses play an important role in the development of periodontitis, and a Th17/Treg imbalance causes the pathogenesis of periodontitis. However, whether vaccination with P. gingivalis can prevent periodontitis through modulation of the Th17/Treg imbalance remains unknown. In this study, mice were subcutaneously vaccinated with formalin-killed P. gingivalis and then orally challenged with P. gingivalis. The vaccination protected the mice from alveolar bone resorption and inflammation. These protective effects might be ascribed to downregulation of Th17 cells and interleukin (IL)-17A production, upregulation of Treg and receptor activator of nuclear factor-kappa B ligand (RANKL)(+)CD4(+)T cells, and IL-10 and transforming growth factor-?1 production, and inhibition of lymphocyte proliferation. Our findings may provide a direction for the development of a vaccine or therapy against periodontitis by alteration of the Th17/Treg imbalance. PMID:25604387

Wang, Linyuan; Guan, Ning; Jin, Ying; Lin, Xiaoping; Gao, Hong

2015-03-01

190

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

191

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

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

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

2006-09-15

192

Gender-Specific Associations of Serum Antibody to Porphyromonas gingivalis and Inflammatory Markers  

Science.gov (United States)

It remains unclear whether serum antibody titer against Porphyromonas gingivalis (Pg) and inflammatory components lead to periodontal deterioration in each gender, as periodontal and systemic status is influenced by gender. The present study investigates the gender-specific probable effects of titer against Pg and inflammatory markers on periodontal health status in a longitudinal study. A retrospective study design was used. At two time points over an 8-year period (in 2003 and 2011), 411 individuals (295 males with a mean age of 57.6 ± 11.2 years and 116 females with a mean age of 59.2 ± 10.3 years) were surveyed. Periodontal status, serum antibody titer against Pg, and high-sensitive C-reactive protein (hsCRP) were evaluated. Poisson regression analyses revealed that the elevated titer against Pg and hsCRP significantly predicted the persistence of periodontal disease 8 years later in females with periodontal disease in 2003. Elevated hsCRP was significantly associated with the incidence of periodontal disease 8 years later in females who were periodontally healthy in 2003. Males had a weaker association among titer against Pg, inflammatory markers, and periodontal disease. These findings suggest that immune response to Pg infection in addition to inflammatory components affects periodontal deterioration in females. PMID:25756052

Furuta, Michiko; Shimazaki, Yoshihiro; Tanaka, Shunichi; Takeuchi, Kenji; Shibata, Yukie; Takeshita, Toru; Nishimura, Fusanori; Yamashita, Yoshihisa

2015-01-01

193

Distribution of Porphyromonas gingivalis fimA genotypes in isolates from subgingival plaque and blood sample during bacteremia / Distribución de los genotipos de fimA en cepas de Porphyromonas gingivalis aisladas de placas subgingivales y de sangre durante bacteriemias  

Scientific Electronic Library Online (English)

Full Text Available SciELO Colombia | Language: English Abstract in spanish Introducción. Porphyromonas gingivalis es el principal agente etiológico de la periodontitis. El gen fimA ha sido relacionado con la virulencia del microorganismo, lo cual sugiere la participación de dicho gen en la capacidad del microorganismo para alcanzar el torrente sanguíneo. Objetivo. Estudiar [...] la distribución de los tipos de fimA de P. gingivalis en muestras de placa subgingival y de sangre obtenidas durante bacteriemias después de raspaje y alisado radicular. Materiales y métodos. Se practicó un alisado radicular a 15 pacientes con periodontitis. Se obtuvieron aislamientos clínicos de P. gingivalis de la placa subgingival y durante la bacteriemia inducida por el procedimiento. Para la genotipificación se utilizó la técnica de reacción en cadena de la polimerasa (PCR) específica para fimA. En los aislamientos no clasificables por PCR se realizó secuenciación del gen fimA. Resultados. Seis pacientes fueron positivos para bacteriemia por P. gingivalis. La distribución de fimA evaluada en 30 aislamientos de placa subgingival y de sangre mostró una mayor frecuencia del fimA tipo II de P. gingivalis. En los aislamientos de placa subgingival, la detección de fimA tipo II fue seguida por Ib, III y IV; sin embargo, en los aislamientos de sangre el tipo II fue seguido por los tipos IV, Ib y III. Conclusión. En los aislamientos de sangre y de placa subgingival de pacientes con periodontitis el fimA más frecuente fue el tipo II; no fue posible correlacionar el tipo de fimA con la bacteriemia inducida por el alisado radicular. Los resultados de la secuenciación del gen fimA no concuerdan con los obtenidos por PCR. Abstract in english Introduction. Porphyromonas gingivalis is considered as a major etiological agent in the onset and progression of chronic destructive periodontitis. Porphyromonus gingivalis fimA type has been correlated to the virulence potential of the strain; therefore this gene could be involved in the ability o [...] f P. gingivalis to reach blood stream. Objective. The classifications of P. gingivalis fimA types will be compared in subgingival plaque and blood samples collected after scaling and root root planing of periodontitis patients. Materials and methods. Fifteen periodontitis patients requiring scaling and root planing were enrolled. P. gingivalis isolates were classed to genotype with fimA type-specific PCR assay. fimA gene was sequenced if the isolate was listed as unclassifiable after PCR technique. Results. Six patients showed positive P. gingivalis bacteremia. The most frequent fimA was fimA type II, followed by Ib, III and IV. In blood strains, type II was followed by IV, Ib and III. Conclusion. Type II was the most frequent genotype in blood samples and in subgingival plaque samples. However, no correlation was found between the frequency of any fimA type with SRP induced bacteremia. P. gingivalis fimA type appears to be conserved within individual patients throughout the times of sample collection. fimA gene sequence results were not in agreement with results of fimA genotyping by PCR.

Paula Juliana, Pérez-Chaparro; Gloria Inés, Lafaurie; Patrice, Gracieux; Vincent, Meuric; Zohreh, Tamanai-Shacoori; Jaime Eduardo, Castellanos; Martine, Bonnaure-Mallet.

2009-06-01

194

Distribución de los genotipos de fimA en cepas de Porphyromonas gingivalis aisladas de placas subgingivales y de sangre durante bacteriemias Distribution of Porphyromonas gingivalis fimA genotypes in isolates from subgingival plaque and blood sample during bacteremia  

Directory of Open Access Journals (Sweden)

Full Text Available Introducción. Porphyromonas gingivalis es el principal agente etiológico de la periodontitis. El gen fimA ha sido relacionado con la virulencia del microorganismo, lo cual sugiere la participación de dicho gen en la capacidad del microorganismo para alcanzar el torrente sanguíneo.
Objetivo. Estudiar la distribución de los tipos de fimA de P. gingivalis en muestras de placa subgingival y de sangre obtenidas durante bacteriemias después de raspaje y alisado radicular.
Materiales y métodos. Se practicó un alisado radicular a 15 pacientes con periodontitis. Se obtuvieron aislamientos clínicos de P. gingivalis de la placa subgingival y durante la bacteriemia inducida por el procedimiento. Para la genotipificación se utilizó la técnica de reacción en cadena de la polimerasa (PCR específica para fimA. En los aislamientos no clasificables por PCR se realizó secuenciación del gen fimA.
Resultados. Seis pacientes fueron positivos para bacteriemia por P. gingivalis. La distribución de fimA evaluada en 30 aislamientos de placa subgingival y de sangre mostró una mayor frecuencia del fimA tipo II de P. gingivalis. En los aislamientos de placa subgingival, la detección de fimA tipo II fue seguida por Ib, III y IV; sin embargo, en los aislamientos de sangre el tipo II fue seguido por los tipos IV, Ib y III.
Conclusión. En los aislamientos de sangre y de placa subgingival de pacientes con periodontitis el fimA más frecuente fue el tipo II; no fue posible correlacionar el tipo de fimA con la bacteriemia inducida por el alisado radicular. Los resultados de la secuenciación del gen fimA no concuerdan con los obtenidos por PCR.Introduction. Porphyromonas gingivalis is considered as a major etiological agent in the onset and progression of chronic destructive periodontitis. Porphyromonus gingivalis fimA type has been correlated to the virulence potential of the strain; therefore this gene could be involved in the ability of P. gingivalis to reach blood stream.
Objective. The classifications of P. gingivalis fimA types will be compared in subgingival plaque and blood samples collected after scaling and root root planing of periodontitis patients.
Materials and methods. Fifteen periodontitis patients requiring scaling and root planing were enrolled. P. gingivalis isolates were classed to genotype with fimA type-specific PCR assay. fimA gene was sequenced if the isolate was listed as unclassifiable after PCR technique.
Results. Six patients showed positive P. gingivalis bacteremia. The most frequent fimA was fimA type II, followed by Ib, III and IV. In blood strains, type II was followed by IV, Ib and III.
Conclusion. Type II was the most frequent genotype in blood samples and in subgingival plaque samples. However, no correlation was found between the frequency of any fimA type with SRP induced bacteremia. P. gingivalis fimA type appears to be conserved within individual patients throughout the times of sample collection. fimA gene sequence results were not in agreement with results of fimA genotyping by PCR.

Zohreh Tamanai-Shacoori

2009-06-01

195

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

196

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

Scientific Electronic Library Online (English)

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

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

2012-04-01

197

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

OpenAIRE

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

198

Porphyromonas gingivalis RgpA-Kgp Proteinase-Adhesin Complexes Penetrate Gingival Tissue and Induce Proinflammatory Cytokines or Apoptosis in a Concentration-Dependent Manner?  

OpenAIRE

The RgpA-Kgp proteinase-adhesin complexes of Porphyromonas gingivalis were observed, using immunostaining, in human gingival tissue associated with periodontitis but not in healthy tissue. The staining pattern suggested a concentration gradient from the subgingival plaque into the subjacent gingival connective tissue. Intense immunostaining was observed in areas displaying gross disturbance of tissue architecture. P. gingivalis cells and the RgpA-Kgp complexes at low concentrations were shown...

O Brien-simpson, Neil M.; Pathirana, Rishi D.; Walker, Glenn D.; Reynolds, Eric C.

2008-01-01

199

Specific In Situ Visualization of Plasma Cells Producing Antibodies against Porphyromonas gingivalis in Gingival Radicular Cyst: Application of the Enzyme-Labeled Antigen Method  

OpenAIRE

The enzyme-labeled antigen method was applied to visualize plasma cells producing antibodies to Porphyromonas gingivalis, flora of the human oral cavity. Antibodies to P. gingivalis have reportedly been detected in sera of patients with periodontitis. Biotinylated bacterial antigens, Ag53, and four gingipain domains (Arg-pro, Arg-hgp, Lys-pro, and Lys-hgp) were prepared by the cell-free protein synthesis system using the wheat germ extract. In paraformaldehyde-fixed frozen sections of rat lym...

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

2011-01-01

200

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

Science.gov (United States)

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

201

Inhibition of peptidase and glycosidase activities of Porphyromonas gingivalis, Bacteroides intermedius and Treponema denticola by plant extracts.  

Science.gov (United States)

Aqueous extracts from 5 plants used widely in Kenya as chewing sticks (mswaki) for the control of oral hygiene were tested for their ability to inhibit extracellular peptidase and glycosidase enzyme activities produced by the periodontopathic bacteria Porphyromonas gingivalis (formerly Bacteroides gingivalis), Bacteroides intermedius and Treponema denticola. The plants studied were Rhus natalensis, Cupressus hisitanica, Sida cordifolia, Olea africana and Euclea divinorum. Protease activities, including glycylprolyl dipeptidase and trypsin-like activities of P. gingivalis, chymotrypsin-like and glycylprolyl dipeptidase activities of B. intermedius and the trypsin-like activity of T. denticola, were particularly affected by extracts from Rhus natalensis and Euclea divinorum. Glycosidase activities were generally less affected with the notable exceptions of the inhibition of beta-mannosidase activity of P. gingivalis by all extracts and the inhibition of neuraminidase activity of T. denticola by Rhus natalensis and Euclea divinorum. Generally, these same proteolytic and glycosidic activities were inhibited by tannic acid and to lesser extents by gallic acid and gallic acid methyl ester. An inhibitory component, present in all extracts, exhibited physical and chemical properties identical to those of tannic acid. The inhibition of these enzyme activities is likely to reduce the virulence of these periodontophathic bacteria and to reduce the rate of dental plaque formation. PMID:1325483

Homer, K A; Manji, F; Beighton, D

1992-05-01

202

Metabolome variations in the Porphyromonas gingivalis vimA mutant during hydrogen peroxide-induced oxidative stress.  

Science.gov (United States)

The adaptability and survival of Porphyromonas gingivalis in the oxidative microenvironment of the periodontal pocket are indispensable for survival and virulence, and are modulated by multiple systems. Among the various genes involved in P. gingivalis oxidative stress resistance, vimA gene is a part of the 6.15-kb locus. To elucidate the role of a P. gingivalis vimA-defective mutant in oxidative stress resistance, we used a global approach to assess the transcriptional profile, to study the unique metabolome variations affecting survival and virulence in an environment typical of the periodontal pocket. A multilayered protection strategy against oxidative stress was noted in P. gingivalis FLL92 with upregulation of detoxifying genes. The duration of oxidative stress was shown to differentially modulate transcription with 94 (87%) genes upregulated twofold during 10 min and 55 (83.3%) in 15 min. Most of the upregulated genes (55%), fell in the hypothetical/unknown/unassigned functional class. Metabolome variation showed reduction in fumarate and formaldehyde, hence resorting to alternative energy generation and maintenance of a reduced metabolic state. There was upregulation of transposases, genes encoding for the metal ion binding protein transport and secretion system. PMID:25055986

McKenzie, R M E; Aruni, W; Johnson, N A; Robles, A; Dou, Y; Henry, L; Boskovic, D S; Fletcher, H M

2015-04-01

203

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

Science.gov (United States)

This study assessed the effect of periodontal therapy on specific serum antibody concentration, expressed as titer, and antibody binding strength, expressed as relative avidity. The immune responses to Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans were investigated. Antibody titer was assayed by enzyme-linked immunosorbent assay (ELISA) and relative avidity was measured by thiocyanate elution in 17 adult periodontitis patients before and after therapy. Immunoglobulin G (IgG) avidities (expressed as thiocyanate molarity) to P. gingivalis increased from 1.01 to 1.38 M (P = 0.05) and IgA titers (expressed as ELISA units [EU]) increased from 89 to 237 EU (P = 0.012). There were no significant changes in avidity to A. actinomycetemcomitans, but the titer of all three immunoglobulin classes increased significantly (P 2 times the control median) or seronegative for P. gingivalis, only patients who were initially seropositive showed a significant increase in antibody avidity (P = 0.026; mean difference, 0.69 M). Patients who were originally seropositive in terms of IgG and IgA titer to P. gingivalis had demonstrably better treatment outcomes in terms of a reduced number of deep pockets and sites which bled on probing (P < 0.05). These findings suggest that periodontal therapy affects the magnitude and quality of the humoral immune response to suspected periodontopathogens, that this effect is dependent on initial serostatus, and that initial serostatus may have a bearing on treatment outcome. PMID:7642270

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

1995-09-01

204

vimA Gene Downstream of recA Is Involved in Virulence Modulation in Porphyromonas gingivalis W83  

OpenAIRE

A 0.9-kb open reading frame encoding a unique 32-kDa protein was identified downstream of the recA gene of Porphyromonas gingivalis. Reverse transcription-PCR and Northern blot analysis showed that both the recA gene and this open reading frame are part of the same transcriptional unit. This cloned fragment was insertionally inactivated using the ermF-ermAM antibiotic resistance cassette to create a defective mutant by allelic exchange. When plated on Brucella blood agar, the mutant strain, d...

Abaibou, Hafid; Chen, Zhuo; Olango, G. Jon; Liu, Yi; Edwards, Jessica; Fletcher, Hansel M.

2001-01-01

205

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

DEFF Research Database (Denmark)

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

Borch, Tanja SkuldbØl; LØbner, Morten

2009-01-01

206

Characterization of wheat germ agglutinin lectin-reactive glycosylated OmpA-like proteins derived from Porphyromonas gingivalis.  

Science.gov (United States)

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

207

Upregulation of heme oxygenase-1 via PI3K/Akt and Nrf-2 signaling pathways mediates the anti-inflammatory activity of Schisandrin in Porphyromonas gingivalis LPS-stimulated macrophages.  

Science.gov (United States)

The lipopolysaccharide (LPS) of Porphyromonas gingivalis is thought to induce periodontitis. In this study, we isolated Schisandrin from the dried fruits of Schisandra chinensis and examined the anti-inflammatory effect of Schisandrin in macrophages stimulated with LPS from P. gingivalis. First, Schisandrin inhibited LPS-induced pro-inflammatory cytokines, including TNF-?, IL-1?, and IL-6. And Schisandrin suppressed the nuclear translocation and activity of NF-?B and phosphorylation of I?B? in LPS-stimulated RAW 264.7 cells. Next, the presence of a selective inhibitor of HO-1 (SnPP) and a siRNA specific for HO-1 inhibited Schisandrin-mediated anti-inflammatory activity. Furthermore, Schisandrin induced HO-1 expression of RAW 264.7 cells through Nrf-2, PI3K/Akt, and ERK activation. Therefore, these results suggest that the anti-inflammatory effects of Schisandrin on P. gingivalis LPS-stimulated RAW 264.7 cells may be due to a reduction of NF-?B activity and induction of the expression of HO-1, leading to TNF-?, IL-1?, and IL-6 down-regulation. PMID:21645546

Park, Sun Young; Park, Da Jung; Kim, Young Hun; Kim, Younghee; Kim, Sun Gun; Shon, Kwang Jae; Choi, Young-Whan; Lee, Sang-Joon

2011-09-30

208

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

Directory of Open Access Journals (Sweden)

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

Lafaurie Gloria

2008-12-01

209

VimA is part of the maturation pathway for the major gingipains of Porphyromonas gingivalis W83.  

Science.gov (United States)

The authors have shown previously that the vimA gene, which is part of the bcp-recA-vimA operon, plays an important role in protease activation in Porphyromonas gingivalis. The gingipain RgpB proenzyme is secreted in the vimA-defective mutant P. gingivalis FLL92. An important question that is raised is whether the vimA gene product could directly interact with the proteases for their activation or regulate a pathway responsible for protease activation. To further study the mechanism(s) of VimA-dependent protease activation, the vimA gene product was further characterized. A 39 kDa protein consistent with the size of the predicted VimA protein was purified. In protein-protein interaction studies, the VimA protein was shown to interact with gingipains RgpA, RgpB and Kgp. Immune sera from mice immunized with P. gingivalis immunoreacted with the purified VimA protein. Taken together, these data suggest an interaction of VimA with the gingipains and further confirm the role of this protein in their regulation or maturation. PMID:17074907

Vanterpool, E; Roy, F; Zhan, W; Sheets, S M; Sangberg, L; Fletcher, H M

2006-11-01

210

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

Directory of Open Access Journals (Sweden)

Full Text Available 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 bone marrow derived macrophages (BMM from young (2-months and aged (1-year and 2-years mice to Pg strain 381. Pg induced robust expression of cytokines; tumor necrosis factor (TNF-?, interleukin (IL-6, and IL-10, chemokines; neutrophil chemoattractant protein (KC, macrophage colony stimulating factor (MCP-1, macrophage inflammatory protein (MIP-1? and regulated upon activation normal T cell expressed and secreted (RANTES, as well as nitric oxide (NO, measured as nitrite, and prostaglandin E2 (PGE2 from BMM of young mice. BMM from the 2-year age group produced significantly less TNF-?, IL-6 and NO in response to Pg as compared with BMM from 2-months and 1-year of age. We did not observe any difference in the levels of IL-1?, IL-10 and PGE2 produced by BMM in response to Pg. BMM from 2-months and 1-year of age produced similar levels of all chemokines measured with the exception of MCP-1, which was reduced in BMM from 1-year of age. BMM from the 2-year group produced significantly less MCP-1 and MIP-1? compared with 2-months and 1-year age groups. No difference in RANTES production was observed between age groups. Employing a Pg attenuated mutant, deficient in major fimbriae (Pg DPG3, we observed reduced ability of the mutant to stimulate inflammatory mediator expression from BMMs as compared to Pg 381, irrespective of age. Taken together these results support senescence as an important facet of the reduced immunological response observed by BMM of aged host to the periodontal pathogen Pg.

Gibson Frank C

2010-11-01

211

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

DEFF Research Database (Denmark)

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

Borch, T S; Holmstrup, Palle

2010-01-01

212

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

Scientific Electronic Library Online (English)

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 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.

2012-02-01

213

Porphyromonas gingivalis trypsin-like protease: a possible natural ligand for the neutrophil formyl peptide receptor.  

Science.gov (United States)

Porphyromonous gingivalis is a periodontopathic Gram-negative anaerobe associated with chronic adult periodontitis. P. gingivalis proteases are considered important virulence factors in the pathogenesis of periodontal diseases. In addition, defective bactericidal activity of neutrophils has also been observed in periodontitis. In this report we describe the effects of trypsin-like protease(s) secreted from P. gingivalis cells on the ligand binding of FMLP receptor on neutrophils. It was observed that trypsin-like protease(s) from P. gingivalis stimulate neutrophils by means of superoxide anion production. Subsequently, the proteases were found to cleave the FMLP receptor protein as evident by direct labeling of the FMLP receptor molecule. These results suggest that trypsin-like protease(s) secreted from P. gingivalis cells contribute to attenuate the bactericidal activity of neutrophils by cleaving the polypeptide chain of the FMLP receptor molecule. The finding that neutrophils after the incubation with P. gingivalis released protease preparation fail to respond to further stimulation by FMLP suggests that P. gingivalis trypsin-like protease(s) may be a possible ligand for the FMLP receptor. PMID:8147895

Lala, A; Amano, A; Sojar, H T; Radel, S J; De Nardin, E

1994-03-30

214

Functional Differences among FimA Variants of Porphyromonas gingivalis and Their Effects on Adhesion to and Invasion of Human Epithelial Cells  

OpenAIRE

Fimbriae of Porphyromonas gingivalis, a periodontopathogen, play an important role in its adhesion to and invasion of host cells. The fimA genes encoding fimbrillin (FimA), a subunit protein of fimbriae, have been classified into five types, types I to V, based on nucleotide sequences. We previously reported that P. gingivalis with type II fimA was strongly associated with adult periodontitis. In the present study, we compared the abilities of recombinant FimA (rFimA) types I to V to adhere t...

Nakagawa, Ichiro; Amano, Atsuo; Kuboniwa, Masae; Nakamura, Takayuki; Kawabata, Shigetada; Hamada, Shigeyuki

2002-01-01

215

The C5a receptor impairs IL-12–dependent clearance of Porphyromonas gingivalis and is required for induction of periodontal bone loss1  

OpenAIRE

The C5a anaphylatoxin receptor (C5aR; CD88) is activated as part of the complement cascade and exerts important inflammatory, antimicrobial and regulatory functions, at least in part, via crosstalk with TLRs. However, the periodontal pathogen Porphyromonas gingivalis can control C5aR activation by generating C5a through its own C5 convertase-like enzymatic activity. Here we show that P. gingivalis uses this mechanism to proactively and selectively inhibit TLR2-induced IL-12p70, whereas the sa...

Liang, Shuang; Krauss, Jennifer L.; Domon, Hisanori; Mcintosh, Megan L.; Hosur, Kavita B.; Qu, Hongchang; Li, Fenge; Tzekou, Apostolia; Lambris, John D.; Hajishengallis, George

2010-01-01

216

Characterization of an adherence and antigenic determinant of the ArgI protease of Porphyromonas gingivalis which is present on multiple gene products.  

OpenAIRE

This study was performed to characterize the antigen(s) recognized by a panel of monoclonal antibodies (MAbs) produced to be specific for Porphyromonas gingivalis whole cells which we had previously shown to bind to epitopes recognized by sera from periodontitis patients. Preliminary data had suggested that the arginine-specific proteases of P. gingivalis (ArgI, ArgIA, and ArgIB) contained the antigenic determinants of four of these antibodies (MAbs 1A1, 2B/H9, 7D5, and 3B1). The location of ...

Curtis, M. A.; Aduse-opoku, J.; Slaney, J. M.; Rangarajan, M.; Booth, V.; Cridland, J.; Shepherd, P.

1996-01-01

217

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.

2005-09-01

218

Ab initio modeling approach towards establishing the structure and docking orientation of the Porphyromonas gingivalis FimA.  

Science.gov (United States)

Porphyromonas gingivalis FimA is a major aetiological agent in periodontal disease development, however, its structure has never been determined. Here, we established the mature P. gingivalis FimA ab initio model of all six FimA variants. We determined the conserved amino acid sequences of each FimA variant and generated mature FimA models. Subsequently, we validated their quality, protein empirical distribution, and radius of gyration. Similarly, structural comparison and topological orientation were elucidated, and the probable protein-protein docking was investigated. We found that the putative mature FimA model is ?-sheet-rich and, likewise, we observed that each mature FimA model has varying levels of structural differences which can be topologically subdivided into the upper, middle, and lower FimA sections. Moreover, we found that the FimA epithelial cell-binding domain (EBD) is structurally conserved within the middle FimA section of all variants and FimA-FimA docking suggests that the FimA EBDs are oriented in opposite and alternating directions of each other. PMID:25424659

Cueno, Marni E; Nagano, Keiji; Imai, Kenichi; Tamura, Muneaki; Yoshimura, Fuminobu; Ochiai, Kuniyasu

2015-02-01

219

The bcp gene in the bcp-recA-vimA-vimE-vimF operon is important in oxidative stress resistance in Porphyromonas gingivalis W83.  

Science.gov (United States)

The ability of Porphyromonas gingivalis to overcome oxidative stress in the inflammatory environment of the periodontal pocket is critical for its survival. We have previously demonstrated that the recA locus, which carries the bacterioferritin co-migratory protein (bcp) gene and has a unique genetic architecture, plays a role in virulence regulation and oxidative stress resistance in P. gingivalis. To further characterize the bcp gene, which was confirmed to be part of the bcp-recA-vimA-vimE-vimF operon, we created a P. gingivalis bcp-defective isogenic mutant (FLL302) by allelic exchange. Compared with the wild-type, FLL302 had a similar growth rate, black pigmentation, ?-hemolysis and UV sensitivity. Although there was no change in the distribution of gingipain activity, there was a 30% reduction in both Arg-X and Lys-X activities in the mutant strain compared with the wild-type. When exposed to 0.25 mm hydrogen peroxide, P. gingivalis FLL302 was more sensitive than the wild-type. In addition, the cloned P. gingivalis bcp gene increased resistance to 0.25 mm hydrogen peroxide in a bcp-defective Escherichia coli mutant. The mutant also demonstrated decreased aerotolerance when compared with the wild-type. Porphyromonas gingivalis FLL302 and the wild-type strain had similar virulence profiles in a mouse model of virulence. These observations suggest that the bcp gene may play a role in oxidative stress resistance but has a decreased functional significance in the pathogenic potential of P. gingivalis. PMID:21214873

Johnson, N A; McKenzie, R M E; Fletcher, H M

2011-02-01

220

Active invasion of Porphyromonas gingivalis and infection-induced complement activation in ApoE-/- mice brains.  

Science.gov (United States)

Periodontal disease is a polymicrobial inflammatory disease that leads to chronic systemic inflammation and direct infiltration of bacteria/bacterial components, which may contribute to the development of Alzheimer's disease. ApoE-/- mice were orally infected (n = 12) with Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia, and Fusobacterium nucleatum as mono- and polymicrobial infections. ApoE-/- mice were sacrificed following 12 and 24 weeks of chronic infection. Bacterial genomic DNA was isolated from all brain tissues except for the F. nucleatum mono-infected group. Polymerase chain reaction was performed using universal 16 s rDNA primers and species-specific primer sets for each organism to determine whether the infecting pathogens accessed the brain. Sequencing amplification products confirmed the invasion of bacteria into the brain during infection. The innate immune responses were detected using antibodies against complement activation products of C3 convertase stage and the membrane attack complex. Molecular methods demonstrated that 6 out of 12 ApoE-/- mice brains contained P. gingivalis genomic DNA at 12 weeks (p = 0.006), and 9 out of 12 at 24 weeks of infection (p = 0.0001). Microglia in both infected and control groups demonstrated strong intracellular labeling with C3 and C9, due to on-going biosynthesis. The pyramidal neurons of the hippocampus in 4 out of 12 infected mice brains demonstrated characteristic opsonization with C3 activation fragments (p = 0.032). These results show that the oral pathogen P. gingivalis was able to access the ApoE-/- mice brain and thereby contributed to complement activation with bystander neuronal injury. PMID:25061055

Poole, Sophie; Singhrao, Sim K; Chukkapalli, Sasanka; Rivera, Mercedes; Velsko, Irina; Kesavalu, Lakshmyya; Crean, StJohn

2015-01-01

221

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

OpenAIRE

Gingipain R2 is a 50 kDa proteinase from the oral pathogenic bacterium Porphyromonas gingivalis. This proteinase, which displays no significant sequence homology to any protein previously analyzed by X-ray crystallography, has been crystallized using the vapor diffusion method. Two different crystal forms were obtained from a solution containing polyethylene glycol (MW 8,000) (space group P2(1)2(1)2(1)) or magnesium sulfate (space group R3) as precipitating agent. Complete diffraction data se...

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

1998-01-01

222

Downregulation of the DNA-Binding Activity of Nuclear Factor-?B p65 Subunit in Porphyromonas gingivalis Fimbria-Induced Tolerance  

OpenAIRE

Porphyromonas gingivalis fimbriae induce high levels of nuclear factor-?B (NF-?B)-dependent cytokine release upon primary but not secondary stimulation of monocytic cells (FimA tolerance). In this study, fimbriae induced Toll-like receptor-mediated activation of both p50 and p65 subunits of NF-?B upon primary cellular activation. However, activation of the transactivating p65 subunit (but not of the transcriptionally inactive p50 subunit) was significantly inhibited in fimbria-restimulated...

Hajishengallis, George; Genco, Robert J.

2004-01-01

223

Modulation of Major Histocompatibility Complex Protein Expression by Human Gamma Interferon Mediated by Cysteine Proteinase-Adhesin Polyproteins of Porphyromonas gingivalis  

OpenAIRE

Cysteine proteinases have been emphasized in the virulence of Porphyromonas gingivalis in chronic periodontitis. These hydrolases may promote the degradation of extracellular matrix proteins and disrupt components of the immune system. In this study it was shown that purified Arg-gingipain and Lys-gingipain inhibited expression of class II major histocompatibility complex (MHC) proteins in response to the stimulation of endothelial cells with human gamma interferon (IFN-?). Treatment with th...

Yun, Peter L. W.; Decarlo, Arthur A.; Hunter, Neil

1999-01-01

224

In situ visualization of plasma cells producing antibodies reactive to Porphyromonas gingivalis in periodontitis: the application of the enzyme-labeled antigen method.  

Science.gov (United States)

Porphyromonas gingivalis is a keystone periodontal pathogen. Histologocally, the gingival tissue in periodontitis shows dense infiltration of plasma cells. However, antigens recognized by antibodies secreted from the immunocytes remain unknown. The enzyme-labeled antigen method was applied to detecting plasma cells producing P. gingivalis-specific antibodies in biopsied gingival tissue of periodontitis. N-terminally biotinylated P. gingivalis antigens, Ag53 and four gingipain domains (Arg-pro, Arg-hgp, Lys-pro and Lys-hgp) were prepared by the cell-free protein synthesis system using wheatgerm extract. With these five labeled proteins as probes, 20 lesions of periodontitis were evaluated. With the AlphaScreen method, antibodies against any one of the five P. gingivalis antigens were detected in 11 (55%) serum samples and 17 (85%) tissue extracts. Using the enzyme-labeled antigen method on paraformaldehyde-fixed frozen sections of gingival tissue, plasma cells were labeled with any one of the five antigens in 17 (94%) of 18 specimens, in which evaluable plasma cells were detected. The positivity rates in periodontitis were significantly higher than those found previously in radicular cysts (20% in sera and 33% in tissue extracts with the AlphaScreen method, and 25% with the enzyme-labeled antigen method). Our findings directly indicate that antibodies reactive to P. gingivalis are locally produced in the gingival lesions, and that inflammatory reactions against P. gingivalis are involved in periodontitis. PMID:24698402

Mizutani, Y; Tsuge, S; Takeda, H; Hasegawa, Y; Shiogama, K; Onouchi, T; Inada, K; Sawasaki, T; Tsutsumi, Y

2014-08-01

225

Porphyromonas gingivalis, Prevotella intermedia and Bacteroides forsythus in plaque subjacent to bridge pontics.  

Science.gov (United States)

This study examined the distribution of P. gingivalis, P. intermedia and B. forsythus in plaque on metallic and porcelain pontics adjacent to healthy and inflamed mucosa. Subpontic plaque was collected from 33 inflamed and 31 healthy sites. Plaque suspension was incubated with specific rabbit antisera to P. gingivalis (FDC 381), P. intermedia (ATCC 25261) and B. forsythus (FDC 335), and the labelled cells disclosed with fluorescein-labelled goat-anti-rabbit IgG by indirect immunofluorescence microscopy. Mean proportions of P. gingivalis, P. intermedia, and B. forsythus at inflamed sites were 0.60+/-0.75, 2.48+/-2.28, and 0.44+/-0.64% respectively, and at healthy sites 0.21+/-0.43, 1.27+/-1.05, and 0.15+/-0.18% respectively. These differences were statistically significant. Almost all sites were positive for P. intermedia, whereas only 12/31 healthy and 21/33 inflamed sites were positive for P. gingivalis. 18/31 healthy and 28/33 inflamed sites were positive for B. forsythus. P. intermedia was recovered in higher proportions from metallic pontics adjacent to inflamed sites (MI) than healthy sites (MH) or porcelain pontics next to inflamed (PI) or healthy sites (PH). P. gingivalis is was recovered in higher proportions from MI than PH. We conclude that both the nature of the pontic material and the health status of the mucosa affect the composition of the associated microbiota. PMID:9565285

Wang, J C; Lai, C H; Listgarten, M A

1998-04-01

226

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

Science.gov (United States)

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

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

2005-07-01

227

Enhancing Specific-Antibody Production to the ragB Vaccine with GITRL That Expand Tfh, IFN-?+ T Cells and Attenuates Porphyromonas gingivalis Infection in Mice  

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The outer membrane protein RagB is one of the major virulence factors of the periodontal pathogen Porphyromonas gingivalis (P. gingivalis). In order to induce protective immune response against P. gingivalis infection, an mGITRL gene-linked ragB DNA vaccine (pIRES-ragB-mGITRL ) was constructed. Six-week-old female BALB/c mice were immunized with pIRES-ragB-mGITRL through intramuscular injection and then challenged by subcutaneous injection in the abdomen with P. gingivalis. RagB-specific antibody-forming cells were evaluated by an Enzyme-linked immunosorbent spot, and specific antibody was determined by enzyme-linked immunosorbent assay. In addition, the frequencies of Tfh and IFN-?+ T cells in spleen were measured using flow cytometer, and the levels of IL-21 and IFN-? mRNA or proteins were detected by real time RT-PCR or ELISA. The data showed that the mGITRL-linked ragB DNA vaccine induced higher levels of RagB-specific IgG in serum and RagB-specific antibody-forming cells in spleen. The frequencies of Tfh and IFN-?+ T cells were obviously expanded in mice immunized by pIRES-ragB-mGITRL compared with other groups (pIRES or pIRES-ragB ). The levels of Tfh and IFN-?+ T cells associated cytokines were also significantly increased in pIRES-ragB-mGITRL group. Therefore, the mice immunized with ragB plus mGITRL showed the stronger resistant to P. gingivalis infection and a significant reduction of the lesion size caused by P. gingivalis infection comparing with other groups. Taken together, our findings demonstrated that intramuscular injection of DNA vaccine ragB together with mGITRL induced protective immune response dramatically by increasing Tfh and IFN-?+ T cells and antibody production to P. gingivalis. PMID:23560053

Su, Zhaoliang; Kong, Fanzhi; Shi, Xiaoju; Tong, Jia; Shen, Pei; Peng, Tianqing; Wang, Shengjun; Xu, Huaxi

2013-01-01

228

Hierarchical gene expression profiles of HUVEC stimulated by different lipid A structures obtained from Porphyromonas gingivalis and Escherichia coli.  

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The ability of lipid A structural variants to elicit unique endothelial cell gene expression was examined by measuring global gene expression profiles in human umbilical cord vein endothelial cells (HUVEC) using Affymetrix full genome chips. Two lipid A structural variants obtained from Porphyromonas gingivalis designated PgLPS(1435/1449) and PgLPS(1690) as well as LPS obtained from Escherichia coli wild type and an E. coli msbB mutant (missing myristic acid in the lipid A) were examined. Each of these lipid A structures has been shown to interact with TLR4; however, PgLPS(1435/1449) and E. coli msbB LPS have been shown to be TLR4 antagonists while PgLPS(1690) and wild-type E. coli LPS are TLR4 agonists. It was found that PgLPS(1435/1449) and PgLPS(1690) as well as E. coli msbB LPS activated a subset of those genes significantly transcribed in response to E. coli wild-type LPS. Furthermore, the subset of genes expressed in response to the different lipid A structural forms were those most significantly activated by wild-type E. coli LPS demonstrating a hierarchy in TLR4-dependent endothelial cell gene activation. A unique gene expression profile for the weak TLR4 agonist PgLPS(1690) was observed and represents a TLR4 hierarchy in endothelial cell gene activation. PMID:17166236

Chen, Casey; Coats, Stephen R; Bumgarner, Roger E; Darveau, Richard P

2007-04-01

229

Inhibition studies of quinazoline-sulfonamide derivatives against the ?-CA (PgiCA) from the pathogenic bacterium, Porphyromonas gingivalis.  

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Abstract Carbonic anhydrases (CAs, EC 4.2.1.1) began to be investigated in detail in pathogenic bacteria, in the search for antibiotics with a novel mechanism of action, since it has been demonstrated that in many bacteria CAs are essential for the life cycle of the organism. The presence of CAs in pathogenic bacteria allows the development of anti-infectives with a new mechanism of action, less explored to date. Here, novel quinazoline derivatives crowned with sulfonamide functionality at position-2 were tested for their ability to inhibit the bacterial ?-CA (PgiCA), identified in the genome of Porphyromonas gingivalis. Six compounds were highly effective, nanomolar inhibitors of the pathogenic enzyme ?-PgCA. Three of them were also highly effective sub-nanomolar inhibitors of the cytosolic human isoform II (hCAII). The best ?-PgCA inhibitor was compound 8c, with a KI of 3.53?nM and selectivity ratio of 24.5 and 24.8 against hCA I and hCA II, respectively. Many of these new compounds showed a high selectivity for bacterial enzyme respect to the mammalian CA isoforms (hCAI and hCAII). These results suggest that sulfonamides with quinazoline scaffold could be considered as suitable candidates for further derivatization to better understand the role of bacterial CAs in pathogenesis. PMID:25407016

Alafeefy, Ahmed M; Ceruso, Mariangela; Al-Tamimi, Abdul-Malek S; Prete, Sonia Del; Supuran, Claudiu T; Capasso, Clemente

2014-11-19

230

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

231

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

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

Takehara Tadamichi

2006-03-01

232

Bactericidal Effect of Extracts and Metabolites of Robinia pseudoacacia L. on Streptococcus mutans and Porphyromonas gingivalis Causing Dental Plaque and Periodontal Inflammatory Diseases.  

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The mouth cavity hosts many types of anaerobic bacteria, including Streptococcus mutans and Porphyromonas gingivalis, which cause periodontal inflammatory diseases and dental caries. The present study was conducted to evaluate the antibacterial potential of extracts of Robinia pseudoacacia and its different fractions, as well as some of its natural compounds against oral pathogens and a nonpathogenic reference bacteria, Escherichia coli. The antibacterial activity of the crude extract and the solvent fractions (hexane, chloroform, ethyl acetate and butanol) of R. pseudoacacia were evaluated against S. mutans, P. gingivalis and E. coli DH5? by standard micro-assay procedure using conventional sterile polystyrene microplates. The results showed that the crude extract was more active against P. gingivalis (100% growth inhibition) than against S. mutans (73% growth inhibition) at 1.8 mg/mL. The chloroform and hexane fractions were active against P. gingivalis, with 91 and 97% growth inhibition, respectively, at 0.2 mg/mL. None of seven natural compounds found in R. pseudoacacia exerted an antibacterial effect on P. gingivalis; however, fisetin and myricetin at 8 µg/mL inhibited the growth of S. mutans by 81% and 86%, respectively. The crude extract of R. pseudoacacia possesses bioactive compounds that could completely control the growth of P. gingivalis. The antibiotic activities of the hexane and chloroform fractions suggest that the active compounds are hydrophobic in nature. The results indicate the effectiveness of the plant in clinical applications for the treatment of dental plaque and periodontal inflammatory diseases and its potential use as disinfectant for various surgical and orthodontic appliances. PMID:25856062

Patra, Jayanta Kumar; Kim, Eun Sil; Oh, Kyounghee; Kim, Hyeon-Jeong; Dhakal, Radhika; Kim, Yangseon; Baek, Kwang-Hyun

2015-01-01

233

Bactericidal Effect of Extracts and Metabolites of Robinia pseudoacacia L. on Streptococcus mutans and Porphyromonas gingivalis Causing Dental Plaque and Periodontal Inflammatory Diseases  

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Full Text Available The mouth cavity hosts many types of anaerobic bacteria, including Streptococcus mutans and Porphyromonas gingivalis, which cause periodontal inflammatory diseases and dental caries. The present study was conducted to evaluate the antibacterial potential of extracts of Robinia pseudoacacia and its different fractions, as well as some of its natural compounds against oral pathogens and a nonpathogenic reference bacteria, Escherichia coli. The antibacterial activity of the crude extract and the solvent fractions (hexane, chloroform, ethyl acetate and butanol of R. pseudoacacia were evaluated against S. mutans, P. gingivalis and E. coli DH5? by standard micro-assay procedure using conventional sterile polystyrene microplates. The results showed that the crude extract was more active against P. gingivalis (100% growth inhibition than against S. mutans (73% growth inhibition at 1.8 mg/mL. The chloroform and hexane fractions were active against P. gingivalis, with 91 and 97% growth inhibition, respectively, at 0.2 mg/mL. None of seven natural compounds found in R. pseudoacacia exerted an antibacterial effect on P. gingivalis; however, fisetin and myricetin at 8 µg/mL inhibited the growth of S. mutans by 81% and 86%, respectively. The crude extract of R. pseudoacacia possesses bioactive compounds that could completely control the growth of P. gingivalis. The antibiotic activities of the hexane and chloroform fractions suggest that the active compounds are hydrophobic in nature. The results indicate the effectiveness of the plant in clinical applications for the treatment of dental plaque and periodontal inflammatory diseases and its potential use as disinfectant for various surgical and orthodontic appliances.

Jayanta Kumar Patra

2015-04-01

234

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 (PSerp-1 produced only a trend toward reductions. Both proteins modified expression of TLR4 and MyD88. In conclusion, aortic plaque growth in ApoE-/- mice increased after angioplasty in mice with chronic oral P. gingivalis infection. 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

235

Porphyromonas gingivalis FimA Fimbriae: Fimbrial Assembly by fimA Alone in the fim Gene Cluster and Differential Antigenicity among fimA Genotypes  

OpenAIRE

The periodontal pathogen Porphyromonas gingivalis colonizes largely through FimA fimbriae, composed of polymerized FimA encoded by fimA. fimA exists as a single copy within the fim gene cluster (fim cluster), which consists of seven genes: fimX, pgmA and fimA-E. Using an expression vector, fimA alone was inserted into a mutant from which the whole fim cluster was deleted, and the resultant complement exhibited a fimbrial structure. Thus, the genes of the fim cluster other than fimA were not e...

Nagano, Keiji; Hasegawa, Yoshiaki; Abiko, Yuki; Yoshida, Yasuo; Murakami, Yukitaka; Yoshimura, Fuminobu

2012-01-01

236

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

OpenAIRE

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

237

Proteins with molecular masses of 50 and 80 kilodaltons encoded by genes downstream from the fimbrilin gene (fimA) are components associated with fimbriae in the oral anaerobe Porphyromonas gingivalis.  

OpenAIRE

Flanking DNA regions of the fimbrilin gene (designated fimA), which encodes the major subunit protein of Porphyromonas (Bacteroides) gingivalis fimbriae, were cloned in several manners from the P. gingivalis chromosome into Escherichia coli by screening with probes derived from a 2.5-kb SacI DNA fragment previously cloned. A total of 10.4 kb of DNA fragments from the P. gingivalis genome was cloned in the pUC plasmid. Expression of the fimA gene and possible flanking genes in the fragments cl...

Yoshimura, F.; Takahashi, Y.; Hibi, E.; Takasawa, T.; Kato, H.; Dickinson, D. P.

1993-01-01

238

Immunization with malondialdehyde-modified low-density lipoprotein (LDL) reduces atherosclerosis in LDL receptor-deficient mice challenged with Porphyromonas gingivalis.  

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Periodontal infections increase the risk of atherosclerotic vascular disease via partly unresolved mechanisms. Of the natural IgM Abs that recognize molecular mimicry on bacterial epitopes and modified lipid and protein structures, IgM directed against oxidized low-density lipoprotein (LDL) is associated with atheroprotective properties. Here, the effect of natural immune responses to malondialdehyde-modified LDL (MDA-LDL) in conferring protection against atherosclerosis, which was accelerated by the major periodontopathogen Porphyromonas gingivalis, was investigated. LDL receptor-deficient (LDLR(-/-)) mice were immunized with mouse MDA-LDL without adjuvant before topical application challenge with live P. gingivalis. Atherosclerosis was analyzed after a high-fat diet, and plasma IgG and IgM Ab levels were measured throughout the study, and the secretion of IL-5, IL-10 and IFN-? in splenocytes stimulated with MDA-LDL was determined. LDLR(-/-) mice immunized with MDA-LDL had elevated IgM and IgG levels to MDA-LDL compared with saline-treated controls. MDA-LDL immunization diminished aortic lipid depositions after challenge with P. gingivalis compared with mice receiving only P. gingivalis challenge. Immunization of LDLR(-/-) mice with homologous MDA-LDL stimulated the production of IL-5, implicating general activation of B-1 cells. Immune responses to MDA-LDL protected from the P. gingivalis-accelerated atherosclerosis. Thus, the linkage between bacterial infectious burden and atherogenesis is suggested to be modulated via natural IgM directed against cross-reactive epitopes on bacteria and modified LDL. PMID:25134521

Turunen, S Pauliina; Kummu, Outi; Wang, Chunguang; Harila, Kirsi; Mattila, Riikka; Sahlman, Marjo; Pussinen, Pirkko J; Hörkkö, Sohvi

2015-05-01

239

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

Science.gov (United States)

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

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

2004-10-01

240

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

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

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

2011-07-01

241

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

242

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

243

Differential quantitative proteomics of Porphyromonas gingivalis by linear ion trap mass spectrometry: Non-label methods comparison, q-values and LOWESS curve fitting  

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Differential analysis of whole cell proteomes by mass spectrometry has largely been applied using various forms of stable isotope labeling. While metabolic stable isotope labeling has been the method of choice, it is often not possible to apply such an approach. Four different label free ways of calculating expression ratios in a classic "two-state" experiment are compared: signal intensity at the peptide level, signal intensity at the protein level, spectral counting at the peptide level, and spectral counting at the protein level. The quantitative data were mined from a dataset of 1245 qualitatively identified proteins, about 56% of the protein encoding open reading frames from Porphyromonas gingivalis, a Gram-negative intracellular pathogen being studied under extracellular and intracellular conditions. Two different control populations were compared against P. gingivalis internalized within a model human target cell line. The q-value statistic, a measure of false discovery rate previously applied to transcription microarrays, was applied to proteomics data. For spectral counting, the most logically consistent estimate of random error came from applying the locally weighted scatter plot smoothing procedure (LOWESS) to the most extreme ratios generated from a control technical replicate, thus setting upper and lower bounds for the region of experimentally observed random error.

Xia, Qiangwei; Wang, Tiansong; Park, Yoonsuk; Lamont, Richard J.; Hackett, Murray

2007-01-01

244

Gingipain RgpB Is Excreted as a Proenzyme in the vimA-Defective Mutant Porphyromonas gingivalis FLL92  

OpenAIRE

We have previously shown that the unique vimA (virulence-modulating) gene could modulate proteolytic activity in Porphyromomas gingivalis. Although a reduction in cysteine protease activity was observed in the vimA-defective mutant, P. gingivalis FLL92, compared to that of the wild-type strain, no changes were seen in the expression of the gingipain genes. This result might suggest posttranscriptional regulation of protease expression. To determine whether there was a defect in the translatio...

Olango, G. Jon; Roy, Francis; Sheets, Shaun M.; Young, Mary K.; Fletcher, Hansel M.

2003-01-01

245

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

246

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.

2010-04-01

247

Anti-HmuY antibodies specifically recognize Porphyromonas gingivalis HmuY protein but not homologous proteins in other periodontopathogens.  

Science.gov (United States)

Given the emerging evidence of an association between periodontal infections and systemic conditions, the search for specific methods to detect the presence of P. gingivalis, a principal etiologic agent in chronic periodontitis, is of high importance. The aim of this study was to characterize antibodies raised against purified P. gingivalis HmuY protein and selected epitopes of the HmuY molecule. Since other periodontopathogens produce homologs of HmuY, we also aimed to characterize responses of antibodies raised against the HmuY protein or its epitopes to the closest homologous proteins from Prevotella intermedia and Tannerella forsythia. Rabbits were immunized with purified HmuY protein or three synthetic, KLH-conjugated peptides, derived from the P. gingivalis HmuY protein. The reactivity of anti-HmuY antibodies with purified proteins or bacteria was determined using Western blotting and ELISA assay. First, we found homologs of P. gingivalis HmuY in P. intermedia (PinO and PinA proteins) and T. forsythia (Tfo protein) and identified corrected nucleotide and amino acid sequences of Tfo. All proteins were overexpressed in E. coli and purified using ion-exchange chromatography, hydrophobic chromatography and gel filtration. We demonstrated that antibodies raised against P. gingivalis HmuY are highly specific to purified HmuY protein and HmuY attached to P. gingivalis cells. No reactivity between P. intermedia and T. forsythia or between purified HmuY homologs from these bacteria and anti-HmuY antibodies was detected. The results obtained in this study demonstrate that P. gingivalis HmuY protein may serve as an antigen for specific determination of serum antibodies raised against this bacterium. PMID:25658942

?miga, Micha?; Bielecki, Marcin; Olczak, Mariusz; Smalley, John W; Olczak, Teresa

2015-01-01

248

Anti-HmuY Antibodies Specifically Recognize Porphyromonas gingivalis HmuY Protein but Not Homologous Proteins in Other Periodontopathogens  

Science.gov (United States)

Given the emerging evidence of an association between periodontal infections and systemic conditions, the search for specific methods to detect the presence of P. gingivalis, a principal etiologic agent in chronic periodontitis, is of high importance. The aim of this study was to characterize antibodies raised against purified P. gingivalis HmuY protein and selected epitopes of the HmuY molecule. Since other periodontopathogens produce homologs of HmuY, we also aimed to characterize responses of antibodies raised against the HmuY protein or its epitopes to the closest homologous proteins from Prevotella intermedia and Tannerella forsythia. Rabbits were immunized with purified HmuY protein or three synthetic, KLH-conjugated peptides, derived from the P. gingivalis HmuY protein. The reactivity of anti-HmuY antibodies with purified proteins or bacteria was determined using Western blotting and ELISA assay. First, we found homologs of P. gingivalis HmuY in P. intermedia (PinO and PinA proteins) and T. forsythia (Tfo protein) and identified corrected nucleotide and amino acid sequences of Tfo. All proteins were overexpressed in E. coli and purified using ion-exchange chromatography, hydrophobic chromatography and gel filtration. We demonstrated that antibodies raised against P. gingivalis HmuY are highly specific to purified HmuY protein and HmuY attached to P. gingivalis cells. No reactivity between P. intermedia and T. forsythia or between purified HmuY homologs from these bacteria and anti-HmuY antibodies was detected. The results obtained in this study demonstrate that P. gingivalis HmuY protein may serve as an antigen for specific determination of serum antibodies raised against this bacterium. PMID:25658942

?miga, Micha?; Bielecki, Marcin; Olczak, Mariusz; Smalley, John W.; Olczak, Teresa

2015-01-01

249

Porphyromonas gingivalis fimbriae  

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Marginal periodontitis is not a homogeneous disease but is rather influenced by an intricate set of host susceptibility differences as well as diversities in virulence among the harbored organisms. It is likely that clonal heterogeneity of subpopulations with both high and low levels of pathogenicity exists among organisms harbored by individuals with negligible, slight, or even severe periodontal destruction. Therefore, specific virulent clones of periodontal pathogens may cause advanced and...

Morten Enersen; Kazuhiko Nakano; Atsuo Amano

2013-01-01

250

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

251

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

252

Gingipain RgpB is excreted as a proenzyme in the vimA-defective mutant Porphyromonas gingivalis FLL92.  

Science.gov (United States)

We have previously shown that the unique vimA (virulence-modulating) gene could modulate proteolytic activity in Porphyromomas gingivalis. Although a reduction in cysteine protease activity was observed in the vimA-defective mutant, P. gingivalis FLL92, compared to that of the wild-type strain, no changes were seen in the expression of the gingipain genes. This result might suggest posttranscriptional regulation of protease expression. To determine whether there was a defect in the translation, transport, or maturation of the gingipains, P. gingivalis FLL92 was further characterized. In contrast to the wild-type strain, a 90% reduction was seen in both Rgp and Kgp protease activities in strain FLL92 during the exponential growth phase. These activities, however, increased to approximately 60% of that of the wild-type strain during stationary phase. Throughout all the growth phases, Rgp and Kgp activities were mostly soluble, in contrast to those of the wild-type strain. Western blot analyses identified unique Rgp- and Kgp-immunoreactive bands in extracellular protein fractions from FLL92 grown to late exponential phase. Also, the RgpB proenzyme was identified in this fraction by mass spectrometry. In addition, in vitro protease activity could be induced by a urea denaturation-renaturation cycle in this fraction. These results indicate that protease activity in P. gingivalis may be growth phase regulated, possibly by multiple mechanisms. Furthermore, the gingipain RgpB is excreted in an inactive form in the vimA mutant. In addition, these results provide the first evidence of posttranslational regulation of protease activity in P. gingivalis and may suggest an important role for the vimA gene in protease activation in this organism. PMID:12819055

Olango, G Jon; Roy, Francis; Sheets, Shaun M; Young, Mary K; Fletcher, Hansel M

2003-07-01

253

Lipopolysaccharides (LPS) of Oral Black-Pigmented Bacteria Induce Tumor Necrosis Factor Production by LPS-Refractory C3H/HeJ Macrophages in a Way Different from That of Salmonella LPS  

OpenAIRE

Some lipopolysaccharide (LPS) preparations from S- or R-form members of the family Enterobacteriaceae and oral black-pigmented bacteria (Porphyromonas gingivalis and Prevotella intermedia) are known to activate LPS-refractory C3H/HeJ macrophages. When contaminating proteins are removed from R-form LPS of Enterobacteriaceae by repurification, however, this ability is lost. In the present study, we investigated the capacity of LPS from P. gingivalis, P. intermedia, Salmonella minnesota, and Sal...

Kirikae, Teruo; Nitta, Toshimasa; Kirikae, Fumiko; Suda, Yasuo; Kusumoto, Shoichi; Qureshi, Nirofer; Nakano, Masayasu

1999-01-01

254

Genome of the pathogen Porphyromonas gingivalis recovered from a biofilm in a hospital sink using a high-throughput single-cell genomics platform.  

Science.gov (United States)

Although biofilms have been shown to be reservoirs of pathogens, our knowledge of the microbial diversity in biofilms within critical areas, such as health care facilities, is limited. Available methods for pathogen identification and strain typing have some inherent restrictions. In particular, culturing will yield only a fraction of the species present, PCR of virulence or marker genes is mainly focused on a handful of known species, and shotgun metagenomics is limited in the ability to detect strain variations. In this study, we present a single-cell genome sequencing approach to address these limitations and demonstrate it by specifically targeting bacterial cells within a complex biofilm from a hospital bathroom sink drain. A newly developed, automated platform was used to generate genomic DNA by the multiple displacement amplification (MDA) technique from hundreds of single cells in parallel. MDA reactions were screened and classified by 16S rRNA gene PCR sequence, which revealed a broad range of bacteria covering 25 different genera representing environmental species, human commensals, and opportunistic human pathogens. Here we focus on the recovery of a nearly complete genome representing a novel strain of the periodontal pathogen Porphyromonas gingivalis (P. gingivalis JCVI SC001) using the single-cell assembly tool SPAdes. Single-cell genomics is becoming an accepted method to capture novel genomes, primarily in the marine and soil environments. Here we show for the first time that it also enables comparative genomic analysis of strain variation in a pathogen captured from complex biofilm samples in a healthcare facility. PMID:23564253

McLean, Jeffrey S; Lombardo, Mary-Jane; Ziegler, Michael G; Novotny, Mark; Yee-Greenbaum, Joyclyn; Badger, Jonathan H; Tesler, Glenn; Nurk, Sergey; Lesin, Valery; Brami, Daniel; Hall, Adam P; Edlund, Anna; Allen, Lisa Z; Durkin, Scott; Reed, Sharon; Torriani, Francesca; Nealson, Kenneth H; Pevzner, Pavel A; Friedman, Robert; Venter, J Craig; Lasken, Roger S

2013-05-01

255

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

256

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

Scientific Electronic Library Online (English)

Full Text Available 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

257

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  

Directory of Open Access Journals (Sweden)

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

258

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

259

Prevalencia de los genotipos fimA II y fimA IV de Porphyromonas gingivalis en un grupo de mujeres mexicanas con diabetes gestacional en la región centro de México / Prevalence of fimA II and fimA IV Porphyromonas gingivalis genotypes in a group of Mexican women with gestational diabetes in the central region of México  

Scientific Electronic Library Online (English)

Full Text Available SciELO Chile | Language: Spanish Abstract in spanish La diabetes gestacional (DG) es una de las complicaciones médicas que más frecuentemente afectan a las mujeres embarazadas; algunos autores reportan una prevalencia entre el 9,7 y el 13,9%. La DG puede ser causa de efectos adversos como: nacimiento pretérmino, macrosomia, nacimiento por cesárea, hip [...] erbilirrubinemia, hipertensión gestacional, así como la predisposición de desarrollar posteriormente diabetes mellitus tipo 2 y síndrome metabólico. La literatura señala la asociación entre los microorganismos presentes en el biofilm subgingival, etiológicos de la inflamación de los tejidos de soporte dentarios y diabetes mellitus. Uno de estos microorganismos, Porphyromonas gingivalis, expresa, entre otros factores de virulencia, una proteína llamada fimbrilina, la cual presenta variaciones genotípicas relacionadas con su capacidad de inducción en la expresión de mediadores inflamatorios; los genotipos fimA II y fimA IV se consideran con mayor capacidad de virulencia y su presencia se ha asociado con la resistencia a la insulina. En este estudio analizamos la prevalencia de los genotipos fimA II y fimA IV en un grupo de mujeres mexicanas de la región central de México con DG, en mujeres con embarazo sin diabetes y mujeres sin embarazo y sin diabetes. Los resultados encontrados muestran una elevada presencia del genotipo fimA II en mujeres con DG (p Abstract in english Gestational diabetes (GD) is one of the most common complications in pregnant women, with some authors reporting prevalence between 9.7% and 13.9%. GD can lead to the following adverse effects: preterm birth, macrosomia, cesarean birth, hyperbilirubinemia, gestational hypertension, and predispositio [...] n to later develop diabetes mellitus type 2 and metabolic syndrome. The literature shows an association between microorganisms in the subgingival biofilm, which produces inflammation of the dental support tissue, and diabetes mellitus. Porphyromonasgingivalis is one of these microorganisms, and among other virulence factors, it expresses a protein called fimbrilin which has genotypic variations related to its ability to induce expression of inflammatory mediators. Genotypes fimA II and fimA IV are considered to have a greater virulence and their presence has been associated with insulin resistance. An analysis is made on the prevalence of genotypes fimA II and fimA IV in a group of women in central region of Mexico with GD, pregnant women without diabetes, and non-pregnant women without diabetes. The results show an elevated presence of genotype fimA II in women with GD (P

Roberto Arturo, García-Reyna; María del Carmen, Terrones Saldivar; Angélica María, Malacara-Rosas; Nicolás, Zaragoza-Velásquez; Alejandro, Rosas-Cabral; Rafael, Gutiérrez Campos.

2014-08-01

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

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P. gingivalis (Pg), a causative agent of chronic generalized periodontitis, has been implicated in promoting cardiovascular disease. Expression of lipoprotein gene PG0717 of Pg strain W83 was found to be transiently upregulated during invasion of human coronary artery endothelial cells (HCAEC), suggesting this protein may be involved in virulence. We characterized the virulence phenotype of a PG0717 deletion mutant of pg W83. There were no differences in the ability of W83?717 to adhere and ...

Reyes, Leticia; Eiler-mcmanis, Eileen; Rodrigues, Paulo H.; Chadda, Amandeep S.; Wallet, Shannon M.; Be?langer, Myriam; Barrett, Amanda G.; Alvarez, Sophie; Akin, Debra; Dunn, William A.; Progulske-fox, Ann

2013-01-01

261

Functional role of interleukin 1 in periodontal disease: induction of interleukin 1 production by Bacteroides gingivalis lipopolysaccharide in peritoneal macrophages from C3H/HeN and C3H/HeJ mice.  

OpenAIRE

Hot phenol-water-extracted lipopolysaccharide (LPS) from Bacteroides gingivalis 381 was purified by Sephadex G-100 chromatography with Tris buffer supplemented with sodium deoxycholate and EDTA (B-LPS). In the present study, B-LPS was examined for its ability to induce interleukin 1 (IL-1) production, a mitogenic response, and macrophage activation in LPS high-responder C3H/HeN and low-responder C3H/HeJ mice. A significant increase in IL-1 production was observed in C3H/HeN and C3H/HeJ perito...

Hanazawa, S.; Nakada, K.; Ohmori, Y.; Miyoshi, T.; Amano, S.; Kitano, S.

1985-01-01

262

Curcumin attenuates cyclooxygenase-2 expression via inhibition of the NF-?B pathway in lipopolysaccharide-stimulated human gingival fibroblasts.  

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Porphyromonas gingivalis lipopolysaccharide (LPS) induces the expression of the cyclooxygenase-2 (COX-2), which contributes to the process of periodontitis. Curcumin, a constituent of turmeric, exhibits anti-inflammatory properties. We have investigated the anti-inflammatory effect of curcumin in human gingival fibroblasts (HGFs) stimulated by P. gingivalis LPS and its mechanism of action. HGFs pretreated with curcumin were stimulated by P. gingivalis LPS. COX-2 mRNA and protein expressions were analysed by real-time PCR and Western blot analysis. Activation of nuclear factor kappa B (NF-?B) was analysed by the NF-?B-dependent luciferase activity and electrophoretic mobility-shift assay (EMSA). Curcumin inhibited COX-2 mRNA and protein synthesis in LPS-stimulated HGFs in a dose-dependent manner. P. gingivalis LPS activated NF-?B-dependent transcription in HGFs, which were also downregulated by pretreatment with curcumin. Therefore, curcumin can inhibit P. gingivalis LPS-induced COX-2 expression, which may be due to the inhibition of the NF-?B pathway. PMID:23494805

Hu, Ping; Huang, Ping; Chen, Min Wei

2013-05-01

263

Actinobacillus Actinomycetemcomitans y Porphyromonas Gingivales como principales patógenos periodontales  

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Full Text Available Entre las bacterias relacionadas con la enfermedad periodontal, existen dos especies más claramente asociadas a esta enfermedad: Actinobacillus actinomycetemcomitans y Porphyromonas gingivalis. Este trabajo es una revisión bibliográfica sobre estos dos patógenos periodontales, mostrando su origen, prevalencia, distribución, transmisión y respuesta al tratamiento periodontal.Among the bacteria related to periodontal disease, there are two species clearly associated to this disease: Actinobacillus actinomycetemcomitans and Porphyromonas gingiva lis. This paper presents a review of the literature regarding this two periodontal pathogens, and showing their origin, prevalence, distribution, transmission and response to periodontal treatment.

A Bascones

2000-09-01

264

Specific cell components of Bacteroides gingivalis mediate binding and degradation of human fibrinogen.  

OpenAIRE

Bacteroides (Porphyromonas) gingivalis, which has been implicated as an etiologic agent in human periodontal diseases, has been shown to bind and degrade human fibrinogen. B. gingivalis strains bind fibrinogen reversibly and with high affinity and bind to a specific region of the fibrinogen molecule that appears to be located between the D and E domains (M. S. Lantz, R. D. Allen, P. Bounelis, L. M. Switalski, and M. Hook, J. Bacteriol. 172:716-726, 1990). We now report that human fibrinogen i...

Lantz, M. S.; Allen, R. D.; Vail, T. A.; Switalski, L. M.; Hook, M.

1991-01-01

265

Bacteroides gingivalis and Bacteroides intermedius recognize different sites on human fibrinogen  

International Nuclear Information System (INIS)

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 fragmentreduced 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

266

Bacteroides gingivalis and Bacteroides intermedius recognize different sites on human fibrinogen  

Energy Technology Data Exchange (ETDEWEB)

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

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

1990-02-01

267

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

OpenAIRE

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 d...

Stefanon Ivanita; Vasquez Elisardo C; Pereira Raquel B; Meyrelles Silvana S

2011-01-01

268

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

Science.gov (United States)

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

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

2013-10-01

269

P. gingivalis in Periodontal Disease and Atherosclerosis – Scenes of Action for Antimicrobial Peptides and Complement  

Science.gov (United States)

According to the NHS, it is estimated that over 50% of the adult population are, to some extent, affected by gum disease and approximately 15% of UK population have been diagnosed with severe periodontitis. Periodontitis, a chronic polymicrobial disease of the gums, causes inflammation in its milder form, whereas in its severe form affects the surrounding tissues and can result in tooth loss. During periodontitis, plaque accumulates and sits between the junctional epithelium and the tooth itself, resulting in inflammation and the formation of a periodontal pocket. An interface is formed directly between the subgingival bacteria and the junctional epithelial cells. Bacterial pathogens commonly associated with periodontal disease are, among others, Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola, together known as the “red complex.” This review will mostly concentrate on the role of P. gingivalis, a Gram-negative anaerobic bacterium and one of the major and most studied contributors of this disease. Because periodontal disease is associated with the development of atherosclerosis, it is important to understand the local immune response to P. gingivalis. Innate immune players, in particular, complement and antimicrobial peptides and their effects with regard to P. gingivalis during periodontitis and in the development of atherosclerosis will be presented. PMID:25713575

Hussain, Mehak; Stover, Cordula M.; Dupont, Aline

2015-01-01

270

Purification and characterisation of recombinant His-tagged RgpB gingipain from Porphymonas gingivalis.  

Science.gov (United States)

Gingipain proteases are important virulence factors from the periodontal pathogen Porphyromonas gingivalis and are the target of many in vitro studies. Due to their close biochemical properties, purification of individual gingipains is difficult and requires multiple chromatographic steps. In this study, we demonstrate that insertion of a hexahistidine affinity tag upstream of a C-terminal outer membrane translocation signal in RgpB gingipain leads to the secretion of a soluble, mature form of RgpB bearing the affinity tag that can easily be purified by nickel-chelating affinity chromatography. The final product obtained high yielding high purity is biochemically indistinguishable from the native RgpB enzyme. PMID:25720118

Veillard, Florian; Potempa, Barbara; Guo, Yonghua; Ksiazek, Miroslaw; Sztukowska, Maryta N; Houston, John A; Koneru, Lahari; Nguyen, Ky-Anh; Potempa, Jan

2015-04-01

271

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

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

Yukie Kogawa

2011-01-01

272

Lipopolysaccharide Endotoxins  

OpenAIRE

Since lipopolysaccharide endotoxins of Gram-negative bacteria were last reviewed in this series in 1990, much has been learned about the assembly and signaling functions of these remarkable glycoconjugates. Lipopolysaccharides typically consist of a hydrophobic domain known as lipid A (or endotoxin), a non-repeating “core” oligosaccharide, and a distal polysaccharide (or O-antigen). The flood of recent genomic data has made it possible to study lipopolysaccharide assembly in diverse Gram-...

Raetz, Christian R. H.; Whitfield, Chris

2001-01-01

273

Diversity of fimbrillin among Porphyromonas gulae clinical isolates from Japanese dogs.  

Science.gov (United States)

Porphyromonas gulae, a gram-negative black-pigmented anaerobe, is a pathogen for periodontitis in dogs. An approximately 41-kDa fimbrial subunit protein (FimA) encoded by fimA is regarded as associated with periodontitis. In the present study, the fimA genes of 17 P. gulae strains were sequenced, and classified into two major types. The generation of phylogenetic trees based on the deduced amino acid sequence of FimA of P. gulae strains along with sequences from several strains of Porphyromonas gingivalis, a major cause of human periodontitis, revealed that the two types of FimA (types A and B) of P. gulae were similar to type I FimA and types II and III FimA of P. gingivalis, respectively. A PCR system for classification was established based on differences in the nucleotide sequences of the fimA genes. Analysis of 115 P. gulae-positive oral swab specimens from dogs revealed that 42.6%, 22.6%, and 26.1% of them contained type A, type B, and both type A and B fimA genes, respectively. Experiments with a mouse abscess model demonstrated that the strains with type B fimA caused significantly greater systemic inflammation than those with type A. These results suggest that the FimA proteins of P. gulae are diverse with two major types and that strains with type B fimA could be more virulent. PMID:22382732

Nomura, Ryota; Shirai, Mitsuyuki; Kato, Yukio; Murakami, Masaru; Nakano, Kazuhiko; Hirai, Norihiko; Mizusawa, Tetsuya; Naka, Shuhei; Yamasaki, Yoshie; Matsumoto-Nakano, Michiyo; Ooshima, Takashi; Asai, Fumitoshi

2012-07-01

274

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

275

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

276

Specific cell components of Bacteroides gingivalis mediate binding and degradation of human fibrinogen  

International Nuclear Information System (INIS)

Bacteroides (Porphyromonas) gingivalis, which has been implicated as an etiologic agent in human periodontal diseases, has been shown to bind and degrade human fibrinogen. B. gingivalis strains bind fibrinogen reversibly and with high affinity and bind to a specific region of the fibrinogen molecule that appears to be located between the D and E domains. The authors now report that human fibrinogen is bound and then degraded by specific B. gingivalis components that appear to be localized at the cell surface. Fibrinogen binding to bacterial cells occurred at 4, 22, and 37 degree C. A functional fibrinogen-binding component (Mr, 150 000) was identified when sodium dodecyl sulfate-solubilized bacteria were fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to nitrocellulose membranes, and probed with 125I-fibrinogen. Fibrinogen degradation did not occur at 4 degree C but did occur at 22 and 37 degree C. When bacteria and iodinated fibrinogen were incubated at 37 degree C, two major fibrinogen fragments (Mr, 97 000 and 50 000) accumulated in incubation mixture supernatant fractions. Two major fibrinogen-degrading components (Mr, 120 000 and 150 000) have been identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in substrate-containing gels. Fibrinogen degradation by the Mr-120 000 and -150 000 proteases was enhanced by reducing agents, completely inhibited by N- ducing agents, completely inhibited by N-?-p-tosyl-L-lysyl chloromethyl ketone, and partially inhibited by n-ethyl maleimide, suggesting that these enzymes are thiol-dependent proteases with trypsinlike substrate specificity. The fibrinogen-binding component could be separated from the fibrinogen-degrading components by selective solubilization of bacteria in sodium deoxycholate

277

Specific cell components of Bacteroides gingivalis mediate binding and degradation of human fibrinogen  

Energy Technology Data Exchange (ETDEWEB)

Bacteroides (Porphyromonas) gingivalis, which has been implicated as an etiologic agent in human periodontal diseases, has been shown to bind and degrade human fibrinogen. B. gingivalis strains bind fibrinogen reversibly and with high affinity and bind to a specific region of the fibrinogen molecule that appears to be located between the D and E domains. The authors now report that human fibrinogen is bound and then degraded by specific B. gingivalis components that appear to be localized at the cell surface. Fibrinogen binding to bacterial cells occurred at 4, 22, and 37{degree}C. A functional fibrinogen-binding component (M{sub r}, 150 000) was identified when sodium dodecyl sulfate-solubilized bacteria were fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to nitrocellulose membranes, and probed with {sup 125}I-fibrinogen. Fibrinogen degradation did not occur at 4{degree}C but did occur at 22 and 37{degree}C. When bacteria and iodinated fibrinogen were incubated at 37{degree}C, two major fibrinogen fragments (M{sub r}, 97 000 and 50 000) accumulated in incubation mixture supernatant fractions. Two major fibrinogen-degrading components (M{sub r}, 120 000 and 150 000) have been identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in substrate-containing gels. Fibrinogen degradation by the M{sub r}-120 000 and -150 000 proteases was enhanced by reducing agents, completely inhibited by N-{alpha}-p-tosyl-L-lysyl chloromethyl ketone, and partially inhibited by n-ethyl maleimide, suggesting that these enzymes are thiol-dependent proteases with trypsinlike substrate specificity. The fibrinogen-binding component could be separated from the fibrinogen-degrading components by selective solubilization of bacteria in sodium deoxycholate.

Lantz, M.S.; Allen, R.D.; Vail, T.A.; Switalski, L.M.; Hook, M. (Univ. of Alabama at Birmingham (USA))

1991-01-01

278

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

OpenAIRE

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...

Alejandra Herrera Herrera; Luis Franco Ospina; Luis Fang; Xed Az Caballero, Antonio D.

2014-01-01

279

Novel inhibitor for prolyl tripeptidyl aminopeptidase from Porphyromonas gingivalis and details of substrate-recognition mechanism.  

Science.gov (United States)

A new inhibitor, H-Ala-Ile-pyrrolidin-2-yl boronic acid, was developed as an inhibitor against prolyl tripeptidyl aminopeptidase with a K(i) value of 88.1 nM. The structure of the prolyl tripeptidyl aminopeptidase complexed with the inhibitor (enzyme-inhibitor complex) was determined at 2.2 A resolution. The inhibitor was bound to the active site through a covalent bond between Ser603 and the boron atom of the inhibitor. This structure should closely mimic the structure of the reaction intermediate between the enzyme and substrate. We previously proposed that two glutamate residues, Glu205 and Glu636, are involved in the recognition of substrates. In order to clarify the function of these glutamate residues in substrate recognition, three mutant enzymes, E205A, E205Q, and E636A were generated by site-directed mutagenesis. The E205A mutant was expressed as an inclusion body. The E205Q mutant was expressed in soluble form, but no activity was detected. Here, the structures of the E636A mutant and its complex with the inhibitor were determined. The inhibitor was located at almost the same position as in the wild-type enzyme-inhibitor complex. The amino group of the inhibitor interacted with Glu205 and the main-chain carbonyl group of Gln203. In addition, a water molecule in the place of Glu636 of the wild-type enzyme interacted with the amino group of the inhibitor. This water molecule was located near the position of Glu636 in the wild-type and formed a hydrogen bond with Gln203. The k(cat)/K(M) values of the E636A mutant toward the two substrates used were smaller than those of the wild-type by two orders of magnitude. The K(i) value of our inhibitor for the E636A mutant was 48.8 microM, which was 554-fold higher than that against the wild-type enzyme. Consequently, it was concluded that Glu205 and Glu636 are significant residues for the N-terminal recognition of a substrate. PMID:18042490

Xu, Yue; Nakajima, Yoshitaka; Ito, Kiyoshi; Zheng, Heng; Oyama, Hiroshi; Heiser, Ulrich; Hoffmann, Torsten; Gärtner, Ulf-Torsten; Demuth, Hans-Ulrich; Yoshimoto, Tadashi

2008-01-18

280

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

OpenAIRE

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 card...

Zeituni, Amir E.; Carrion, Julio; Cutler, Christopher W.

2010-01-01

281

Growth inhibitory effects of endotoxins from Bacteroides gingivalis and intermedius on human gingival fibroblasts in vitro  

International Nuclear Information System (INIS)

Purified endotoxin or lipopolysaccharide from Bacteroides gingivalis and Bacteroides intermedius caused a similar dose-dependent inhibition of growth of cultured human gingival fibroblasts as determined by 3H-thymidine incorporation and direct cell count. Approximately 200 micrograms/ml endotoxin caused a 50% reduction in 3H-thymidine uptake of logarithmically growing cells. Inhibition of growth was similar in cultures of fibroblasts derived from either healthy or diseased human gingiva. When examining the change in cell number with time of exposure in culture, the rate of proliferation was significantly suppressed during the logarithmic phase of growth. However, the cells recovered so that the rate of proliferation, although reduced, was sufficient to produce a cell density similar to the control cells with prolonged culture. The endotoxins were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The profiles of the Bacteroides endotoxins were different. B. gingivalis endotoxin showed a wide range of distinct bands indicating a heterogeneous distribution of molecular species. Endotoxin from B. intermedius exhibited a few discrete low molecular weight bands, but the majority of the lipopolysaccharides electrophoresed as a diffuse band of high molecular weight material. The apparent heterogeneity of the two Bacteroides endotoxins and the similarity in growth inhibitory capacity suggest that growth inhibitory effects of thesest that growth inhibitory effects of these substances cannot be attributed to any polysaccharide species of endotoxin

282

Distribution and molecular characterization of Porphyromonas gulae carrying a new fimA genotype.  

Science.gov (United States)

Porphyromonas gulae is a gram-negative black-pigmented anaerobe which is known to be a pathogen for periodontitis in dogs. Approximately 41kDa filamentous appendages on the cell surface (FimA) encoded by the fimA gene are regarded as important factors associated with periodontitis. The fimA genotype was classified into two major types and strains in type B were shown to be more virulent than those in type A. In the present study, we characterized a strain with a novel fimA genotype and designated it as type C. The putative amino acid sequence was shown to be similar to the genotype IV fimA of Porphyromonas gingivalis, a major pathogen of human periodontitis. Analyses using an oral squamous cell carcinoma cell line derived from tongue primary lesions revealed that the type C strain inhibited proliferation and scratch closure more than genotype A and B strains. In addition, experiments using a mouse abscess model demonstrated that the type C strain could induce much higher systemic inflammation when compared with strains of the other genotypes. Furthermore, molecular analyses of oral swab specimens collected from dogs demonstrated that the detection frequencies of P. gulae and the genotype C in the periodontitis group were significantly higher than those in the periodontally healthy group. These results suggest that FimA of P. gulae is diverse with the virulence of genotype C strains the highest and that molecular identification of genotype C P. gulae could be a possible useful marker for identifying dogs at high risk of developing periodontitis. PMID:22877518

Yamasaki, Yoshie; Nomura, Ryota; Nakano, Kazuhiko; Inaba, Hiroaki; Kuboniwa, Masae; Hirai, Norihiko; Shirai, Mitsuyuki; Kato, Yukio; Murakami, Masaru; Naka, Shuhei; Iwai, Soichi; Matsumoto-Nakano, Michiyo; Ooshima, Takashi; Amano, Atsuo; Asai, Fumitoshi

2012-12-28

283

Protein kinase A enhances lipopolysaccharide-induced IL-6, IL-8, and PGE2 production by human gingival fibroblasts  

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Full Text Available Abstract Objective Periodontal disease is accompanied by inflammation of the gingiva and destruction of periodontal tissues, leading to alveolar bone loss in severe clinical cases. Interleukin (IL-6, IL-8, and the chemical mediator prostaglandin E2 (PGE2 are known to play important roles in inflammatory responses and tissue degradation. Recently, we reported that the protein kinase A (PKA inhibitor H-89 suppresses lipopolysaccharide (LPS-induced IL-8 production by human gingival fibroblasts (HGFs. In the present study, the relevance of the PKA activity and two PKA-activating drugs, aminophylline and adrenaline, to LPS-induced inflammatory cytokines (IL-6 and IL-8 and PGE2 by HGFs were examined. Methods HGFs were treated with LPS from Porphyromonas gingivalis and H-89, the cAMP analog dibutyryl cyclic AMP (dbcAMP, aminophylline, or adrenaline. After 24 h, IL-6, IL-8, and PGE2 levels were evaluated by ELISA. Results H-89 did not affect LPS-induced IL-6 production, but suppressed IL-8 and PGE2 production. In contrast, dbcAMP significantly increased LPS-induced IL-6, IL-8, and PGE2 production. Up to 10 ?g/ml of aminophylline did not affect LPS-induced IL-6, IL-8, or PGE2 production, but they were significantly increased at 100 ?g/ml. Similarly, 0.01 ?g/ml of adrenaline did not affect LPS-induced IL-6, IL-8, or PGE2 production, but they were significantly increased at concentrations of 0.1 and 1 ?g/ml. In the absence of LPS, H-89, dbcAMP, aminophylline, and adrenaline had no relevance to IL-6, IL-8, or PGE2 production. Conclusion These results suggest that the PKA pathway, and also PKA-activating drugs, enhance LPS-induced IL-6, IL-8, and PGE2 production by HGFs. However, aminophylline may not have an effect on the production of these molecules at concentrations used in clinical settings (8 to 20 ?g/ml in serum. These results suggest that aminophylline does not affect inflammatory responses in periodontal disease.

Ara Toshiaki

2012-03-01

284

Assessment of antibacterial effect of cinnamon on growth of porphyromons gingivalis from chronic periodontitis patients with deep pockets (in vitro  

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Full Text Available   Background and Aims : Antibiotics are commonly used for controlling the growth of porphyromons gingivalis (P.g which is one of the most important etiologic factors in the periodontal diseases. Different side effects of synthetics and chemical drugs such as increasing the drug resistancy in the human pathogens have led to study on the herbal antibacterial effect. The aim of this study was to evaluate the antibacterial effect of cinnamon on the growth of porphyromons gingivalis in chronic periodontitis patients with deep pockets.   Materials and Methods: In this experimental study, samples were provided from patients having pockets. After culturing the microorganism and diagnosis of P.g by gram staining and biochemical tests, cinnamon in different concentrations (10, 50, 100, 250, 500, 750 and 1500 mg/ml with oil solvent were prepared and placed by disks in the cultures medium. Positive controls were amoxicillin, metronidazole, ciprofloxacin, amikacin and gentamycin . Oil was negative control. Then the plates were incubated for 24 hours in 37 0 C and then non-growth halos by disk diffusion method, MIC (Minimum Inhibitory Concentration and MBC (Minimum Bactericidal Concentration were determined. Data were analyzed using One-way ANOVA test.   Results: The results showed that the cinnamon at the concentration of MIC=750 mg/ml had the inhibitory effects of bacteria and at the concentration of MIC=1500 mg/ml had killing effect. However, this antibacterial effect compared with commonly used antibiotics (amoxicillin, metronidazole, was much weaker (P<0.001.   Conclusion: Cinnamon showed an antimicrobial effect on porphyromonas gingivalis in chronic periodontitis patients with deep pockets.

Babak Amoian

2014-04-01

285

Subgingival Microbial Profile And Production Of Proinflammatory Cytokines In Chronic Periodontitis  

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Full Text Available ????????? ????? ?????????? ?????? ? ?????????? ? ????? ? ????????????? ???????? ??? ??????????? ??????????? ? ??????????? ?? ??????? ??????? ? ???????????? ? ???? ?????? ???? ????? ??????????? ?????????? ?????????????? ?????????? ? ??????????? ???????????????? ???????? ? ?????????????? ?????. ???????????, ??? ??????????? ?????????? ??????????? ????? ? 80% ????????? ???????? ????????, ? ?????? ? ??????????????? ???????????? ??????????? - ? ???????? 1-1.5%. ?????????, ??? ????????? ????? ??????????? ??????????? ??????????????, ?????? ????????? ? ?????????? ?????? ??????????, ??? ?????? ??????????? ????????????????? ??????????????? ? ?????????????? ????????? ????????? ?????? ???????? ???? ? ????????? ? ???????????????? ???????????. ????? ?????????????? ????? ????????, ?????????????? ? ?????????????? ?????, ??? ???? - Porphyromonas gingivalis (PG, Treponema denticola (TD, Tannerella forsythia (TF ?????? ??????? ? ????????? ? ????????????????? ???????????. ?????????????????? ???????????? ???? ????????? ??????? Porphyromonas gingivalis ???????? ?????????????? ???????, ????????????? ????? ? ??????????? ???????????? ???????????. ? ?????? ??????? ??????????? Aggregatibacter actinomycetemcomitans ? ????????? ? ??????????? ???????????? ????? ??????? ? ???????? ??????????? ? ???????????? ??????????? ???????????????? ?????. ?????????? ?????????? ?????????????? ????????? ? ??????????????? ???????????? ????? ???????????? ?????? ???????? ?????????? ?? ??????? ?????????, ????????? ? ??????????????? ???????? ?????????? ? ?????????? (?????????? ????? ????????????? ???????? ? ??????? ????????? ??????????? ??????????? ????????? ?????????? ?????????. ????????? ??????, ????????? ? ??????????, ??????????, ??? ??????????? ???????????? ??????????????? ? ?????????????? ????? ????? ????? ? ??????????? ???????? ???????????????? ????????? ? ???????????? ???????? ? ? ???????????? ??????, ?????? ??????? ??????????? ? ????????????? ???????, ??? ?????? ????????? ? ????????????? ??????? ??????. ? ?????????? ?? ??????? ?????? ?????????????? ? ?????????? ??????? ?????????????????? ? ?????? ? ????????? ???????? ??????? ? ??????????? ??????????????? ???????????. ??? ?????????? ?????????? ???????? ?????????? ???????????????? ????????? ??? ?????-???

Dosseva-Panova Velichka T.

2014-09-01

286

Distinctive pathways characterize A. actinomycetemcomitans and P. gingivalis.  

Science.gov (United States)

The study aimed to compare the molecular mechanism of Porphuromonas gingivalis (P. gingivalis) and Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans). With microarray dataset (GSE9723) from Gene Expression Omnibus, differentially expressed genes (DEGs) were identified comparing normal cell samples with A. actinomycetemcomitans-infected and P. gingivalis-infected periodontitis cell samples, respectively (|logFC| > 1, p value Cytoscape, followed by pathway enrichment analysis (p value <0.05) using FISHER hyper-geometric algorithm and calculation of pathway alter scores using LIMMA. Totally, 839 DEGs and 251 DEGs were screened for A. actinomycetemcomitans and P. gingivalis, respectively. A. actinomycetemcomitans-related network had lower ASPL, degree and EC but higher CC and TC (p < 0.01), while P. gingivalis-related network had lower EC but higher CC and BC (p < 0.01) compared to background network. P. gingivalis-related network had lower ASPL, degree and EC, but higher CC than the A. actinomycetemcomitans-related network (p < 0.05). A. actinomycetemcomitans was associated with the pathways relating to endothelial cells function, while neuroactive ligand-receptor interaction and MAPK pathways were important for P. gingivalis, which had higher alter scores in hematopoietic cell lineage, hypertrophic cardiomyopathy and arrhythmogenic right ventricular cardiomyopathy pathways than A. actinomycetemcomitans. Genes and pathways of the two pathogens were distinctive. The findings aided in preventing and treating relevant diseases. PMID:25351486

Lv, Jing; Zhu, Yi-Xin; Liu, Ying-Qun; Xue, Xin

2015-02-01

287

Hypoxia augments lipopolysaccharide-induced cytokine expression in periodontal ligament cells.  

Science.gov (United States)

Periodontitis is a chronic inflammatory disease characterized by the destruction of tooth supporting tissues. Hypoxia, the mainly changes of the plateau environment, can induce severe periodontitis by animal experiments. There is, however, very little information on hypoxia and lipopolysaccharide (LPS) induced cytokine expression in periodontal ligament (PDL) cells. In this article, we characterized hypoxia or P. gingivalis lipopolysaccharide (Pg LPS) induced tumor necrosis factor alpha (TNF-?), interleukin (IL)-1?, and IL-6 expression by human periodontal ligament (hPDL) cells. We found that hypoxia augmented Pg LPS induced TNF-?, IL-1?, and IL-6 expression in hPDL cells. We also demonstrated that nuclear factor kappa B pathway was involved in hypoxia augmenting Pg LPS induced cytokine expression in hPDL cells. Thus, our results suggest that the hypoxic environment may enhance the immune function of hPDL cells that is induced by Pg LPS. PMID:24609838

Jian, Congxiang; Li, Chenjun; Ren, Yu; He, Yong; Li, Yunming; Feng, Xiaodan; Zhang, Gang; Tan, Yinghui

2014-10-01

288

The atypical lipopolysaccharide of Francisella  

Science.gov (United States)

Bacterial lipopolysaccharides (LPSs) are ubiquitous molecules that are prominent components of the outer membranes of most gram-negative bacteria. Genetic and structural characterizations of Francisella LPS have revealed substantial differences when compared to more commonly studied LPSs of the Enterobacteriaceae. This review discusses both the general characteristics and the unusual features of Francisella LPS. PMID:23916469

Okan, Nihal A.; Kasper, Dennis L.

2013-01-01

289

Antigenicity of Helicobacter pylori lipopolysaccharides.  

OpenAIRE

An investigation of the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) profiles of lipopolysaccharides (LPSs) extracted from seven strains of Helicobacter pylori revealed that these molecules were silver stainable and exhibited a high degree of variability in their patterns. Two strains synthesized a variety of sizes of LPS molecules such that fractionation by SDS-PAGE resulted in a stepwise gradation of bands which extended from the top to the bottom of the silver-stain...

Mills, S. D.; Kurjanczyk, L. A.; Penner, J. L.

1992-01-01

290

Role of TLR2-dependent IL-10 production in the inhibition of the initial IFN-? T cell response to Porphyromonas gingivalis.  

Science.gov (United States)

P.g., a Gram-negative bacterium, is one of the main etiological agents of the chronic inflammatory disease, periodontitis. Disease progression is thought to occur as a result of an inadequate immune response, which although happens locally, can also occur distally as a result of the dissemination of P.g. into the circulation. As IL-10 and TLR2 are pivotal molecules in the immune response that P.g. elicits, we hypothesized that TLR2-mediated IL-10 production, following the initial systemic exposure to P.g., inhibits the IFN-? T cell response. To address this hypothesis, mice were primed with P.g., and the types of cells producing IL-10 and the capacity of T cells to produce IFN-? following blocking or neutralization of IL-10 were assessed. Our results showed that upon initial encounter with P.g., splenic T cells and CD11b(+) cells produce IL-10, which when neutralized, resulted in a substantial increase in IFN-? production by T cells. Furthermore, IL-10 production was dependent on TLR2/1 signaling, partly in response to the major surface protein, FimA of P.g. In addition, P.g. stimulation resulted in the up-regulation of PD-1 and its ligand PD-L1 on CD4 T cells and CD11b(+) cells, respectively. Up-regulation of PD-1 was partially dependent on IL-10 but independent of TLR2 or FimA. These results highlight the role of IL-10 in inhibiting T cell responses to the initial systemic P.g. exposure and suggest multiple inhibitory mechanisms potentially used by P.g. to evade the host's immune response, thus allowing its persistence in the host. PMID:23077245

Gaddis, Dalia E; Maynard, Craig L; Weaver, Casey T; Michalek, Suzanne M; Katz, Jannet

2013-01-01

291

HIV-1 Reactivation Induced by the Periodontal Pathogens Fusobacterium nucleatum and Porphyromonas gingivalis Involves Toll-Like Receptor 4 and 9 Activation in Monocytes/Macrophages?  

OpenAIRE

Although oral coinfections (e.g., periodontal disease) are highly prevalent in human immunodeficiency virus type 1-positive (HIV-1+) patients and appear to positively correlate with viral load levels, the potential for oral bacteria to induce HIV-1 reactivation in latently infected cells has received little attention. We showed that HIV-1 long terminal repeat (LTR) promoter activation can be induced by periodontopathogens in monocytes/macrophages; nevertheless, the mechanisms involved in this...

Gonza?lez, Octavio A.; Li, Mengtao; Ebersole, Jeffrey L.; Huang, Chifu B.

2010-01-01

292

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  

OpenAIRE

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 model...

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

293

Bacteroides gingivalis-Actinomyces viscosus cohesive interactions as measured by a quantitative binding assay  

International Nuclear Information System (INIS)

There is limited evidence, mostly indirect, to suggest that the adherence of Bacteroides gingivalis to teeth may be enhanced by the presence of gram-positive dental plaque bacteria like Actinomyces viscosus. The purpose of this study was to carry out direct quantitative assessments of the cohesion of B gingivalis and A. viscosus by using an in vitro assay modeled on the natural sequence in which these two species colonize the teeth. The assay allowed comparisons to be made of the adherence of 3H-labeled B. gingivalis 2561 and 381 to saliva-coated hydroxyapatite beads (S-HA) and A. viscosus WVU627- or T14V-coated S-HA (actinobeads) in equilibrium and kinetics binding studies. A series of preliminary binding studies with 3H-labeled A. viscosus and parallel studies by scanning electron microscopy with unlabeled A. viscosus were conducted to establish a protocol by which actinobeads suitable for subsequent Bacteroides adherence experiments could be prepared. By scanning electron microscopy, the actinobeads had only small gaps of exposed S-HA between essentially irreversibly bound A. viscosus cells. Furthermore, B. gingivalis cells appeared to bind preferentially to the Actinomyces cells instead of the exposed S-HA. B. gingivalis binding to both S-HA and actinobeads was saturable with at least 2 X 10(9) to 3 X 10(9) cells per ml, and equilibrium with saturating concentrations was reached within 10 to 20 min. B. gingivalis always bound in greater numbers to the as always bound in greater numbers to the actinobeads than to S-HA. These findings provide direct measurements supporting the concept that cohesion with dental plaque bacteria like A. viscosus may foster the establishment of B. gingivalis on teeth by enhancing its adherence

294

LPS-induced Inflammatory Response after Therapy of Aggressive Periodontitis  

OpenAIRE

We have reported a lipopolysaccharide (LPS)-induced hyper-inflammatory response in localized aggressive periodontitis (LAP). It is unknown whether treatment is able to modulate this LPS responsiveness. Fifty-nine individuals with LAP were treated by mechanical debridement and systemic antibiotics. Clinical parameters and cyto/chemokine responsiveness of whole blood stimulated with Porphyromonas gingivalis or Escherichia coli LPS were monitored at baseline and 3, 6, and 12 months post-treatmen...

Shaddox, L. M.; Gonc?alves, P. F.; Vovk, A.; Allin, N.; Huang, H.; Hou, W.; Aukhil, I.; Wallet, S. M.

2013-01-01

295

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

OpenAIRE

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)-s...

Marquis Annie; Genovese Salvatore; Epifano Francesco; Grenier Daniel

2012-01-01

296

Maxillary osteomyelitis due to Halicephalobus gingivalis and fatal dissemination in a horse / Osteomielitis maxilar debido a Halicephalobus gingivalis y diseminación fatal en un caballo  

Scientific Electronic Library Online (English)

Full Text Available SciELO Chile | Language: English Abstract in spanish En la presente comunicación se expone un caso de infestación parasitaria poco habitual causada por Halicephalobus gingivalis, cuya manifestación principal fue osteomielitis del hueso maxilar. El caballo mostraba inicialmente inflamación y dolor en la región de la cresta facial derecha. Las radiograf [...] ías demostraron la presencia de osteolisis y ensanchamiento de la cresta facial. La biopsia del hueso mostraba inflamación granulomatosa y un gran número de larvas del nematodo. El caballo fue tratado con ivermectina. Inicialmente mejoraron los signos clínicos, pero dos meses y medio después el caballo desarrolló uveítis y fallo renal, por lo que fue eutanasiado. El estudio anatomopatológico mostró múltiples granulomas parasitarios en los riñones y en la úvea. La infección por Halicephalobus gingivalis es poco frecuente en caballos y personas aunque presenta una distribución mundial. De acuerdo con los autores esta es la primera vez que se describe dicha infestación en un équido en España. Abstract in english This study reports a rare case of maxillary osteomyelitis in a horse caused by Halicephalobus gingivalis. The horse presented inflammation and pain in the region of the right facial crest and the radiographs detected osteolysis and widening of the facial crest. The biopsy revealed a granulomatous in [...] flammation and a large amount of parasite larvae. The horse was treated with ivermectin but it developed uveitis and renal insufficiency 2.5 months later and was euthanised. The anatomopathological study found multiple parasitic granulomas in the kidneys and uveal tract. H. gingivalis is an infrequent infection in horses and people, and it has a worldwide distribution. To the best of our knowledge this is the first report of H. gingivalis infection in an equid to be diagnosed in Spain.

LA, Gracia-Calvo; M, Martín-Cuervo; ME, Durán; V, Vieítez; F, Serrano; J, Jiménez; LJ, Ezquerra.

297

Draft genome sequences of 26 porphyromonas strains isolated from the canine oral microbiome.  

Science.gov (United States)

We present the draft genome sequences for 26 strains of Porphyromonas (P. canoris, P. gulae, P. cangingavalis, P. macacae, and 7 unidentified) and an unidentified member of the Porphyromonadaceae family. All of these strains were isolated from the canine oral cavity, from dogs with and without early periodontal disease. PMID:25858832

Coil, David A; Alexiev, Alexandra; Wallis, Corrin; O'Flynn, Ciaran; Deusch, Oliver; Davis, Ian; Horsfall, Alexander; Kirkwood, Nicola; Jospin, Guillaume; Eisen, Jonathan A; Harris, Stephen; Darling, Aaron E

2015-01-01

298

Hypoxia aggravates lipopolysaccharide-induced lung injury  

Science.gov (United States)

The animal model of inflammatory response induced by intratracheal application of lipopolysaccharide includes many typical features of acute lung injury or the acute respiratory distress syndrome. A number of experimental investigations have been performed to characterize the nature of this injury more effectively. In inflammatory conditions, hypoxia occurs frequently before and in parallel with pulmonary and non-pulmonary pathological events. This current study was designed to examine the in vivo effect of hypoxia as a potentially aggravating condition in endotoxin-induced lung injury. Lipopolysaccharide, 150 µg, was instilled intratracheally into rat lungs, and thereafter animals were exposed to either normoxia or hypoxia (10% oxygen). Lungs were collected 2, 4, 6 and 8 h later. Inflammatory response and tissue damage were evaluated by quantitative analysis of inflammatory cells and mediators, surfactant protein and vascular permeability. A significantly enhanced neutrophil recruitment was seen in lipopolysaccharide-animals exposed to hypoxia compared to lipopolysaccharide-animals under normoxia. This increased neutrophil accumulation was triggered by inflammatory mediators such as tumour necrosis factor-? and macrophage inflammatory protein-1?, secreted by alveolar macrophages. Determination of vascular permeability and surfactant protein-B showed enhanced concentrations in lipopolysaccharide-lungs exposed to hypoxia, which was absent in animals previously alveolar macrophage-depleted. This study demonstrates that hypoxia aggravates lipopolysaccharide injury and therefore represents a second hit injury. The additional hypoxia-induced inflammatory reaction seems to be predominantly localized in the respiratory compartment, underlining the compartmentalized nature of the inflammatory response. PMID:15996189

Vuichard, D; Ganter, M T; Schimmer, R C; Suter, D; Booy, C; Reyes, L; Pasch, T; Beck-Schimmer, B

2005-01-01

299

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

Energy Technology Data Exchange (ETDEWEB)

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

300

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)

301

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

OpenAIRE

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

302

Lipopolysaccharide banding patterns of Neisseria meningitidis and Neisseria gonorrhoeae.  

OpenAIRE

The lipopolysaccharides of Neisseria meningitidis and Neisseria gonorrhoeae were examined by electrophoresis after whole-cell lysis and proteinase K digestion. The banding patterns observed from clinical isolates and laboratory strains demonstrated lipopolysaccharide which included a small number of smooth high-molecular-weight molecules as well as the previously reported lower-molecular-weight rough lipopolysaccharide.

Parr, T. R.; Bryan, L. E.

1984-01-01

303

Purification and properties of hemagglutinin from culture supernatant of Bacteroides gingivalis.  

OpenAIRE

The hemagglutinating factor (hemagglutinin) of Bacteroides gingivalis was prepared from the supernatant of a 5-day diffusate broth culture by ammonium sulfate precipitation and column chromatography with a hydrophobic column of Phenyl-Sepharose CL-4B, DEAE-Sephadex A-50, and Sephadex G-100 gel filtration. The hemagglutinating activity of the preparation was 53.3 times higher than that of ammonium sulfate precipitate. In electron microphotographs, hemagglutinin appears to have a vesicle or tub...

Okuda, K.; Yamamoto, A.; Naito, Y.; Takazoe, I.; Slots, J.; Genco, R. J.

1986-01-01

304

Lysozyme-mediated aggregation and lysis of the periodontal microorganism Capnocytophaga gingivalis 2010.  

OpenAIRE

The ability of lysozyme to aggregate and lyse the gram-negative capnophilic periodontal microorganism Capnocytophaga gingivalis 2010 was monitored optically at 540 nm. Both hen egg white and chromatographically purified human lysozymes had significant but similar aggregation potentials for both logarithmic- and stationary-phase bacteria. In general, an increase in enzyme concentration resulted in a graded increase in both the initial and maximum changes in turbidity which occurred during the ...

Iacono, V. J.; Zove, S. M.; Grossbard, B. L.; Pollock, J. J.; Fine, D. H.; Greene, L. S.

1985-01-01

305

Lipopolysaccharide (LPS) stimulation of fungal secondary metabolism  

OpenAIRE

We report on a preliminary investigation of the use the Gram-negative bacterial cell wall constituent lipopolysaccharide (LPS) as a natural chemical cue to stimulate and alter the expression of fungal secondary metabolism. Integrated high-throughput micro-cultivation and micro-analysis methods determined that 6 of 40 (15%) of fungi tested responded to an optimal exposure to LPS (0.6 ng/mL) by activating, enhancing or accelerating secondary metabolite production. To explore the possible mecha...

Khalil, Zeinab G.; Kalansuriya, Pabasara; Capon, Robert J.

2014-01-01

306

Lipopolysaccharide Complexes with Pasteurella haemolytica Leukotoxin  

OpenAIRE

The presence of lipopolysaccharide (LPS) in gram-negative bacterial repeats-in-toxin (RTX) toxin preparations, as well as the harsh conditions required to remove it, suggests that LPS may complex with RTX toxins. Concentrated culture supernatant (CCS) preparations of the RTX toxin Pasteurella haemolytica leukotoxin (LKT) contained LKT and LPS as the most prominent components, with LKT and LPS constituting ?30 and 50% of the density of the silver-stained fraction on sodium dodecyl sulfate-po...

Li, Jun; Clinkenbeard, Kenneth D.

1999-01-01

307

Affinity purification of bovine antibodies to Brucella abortus Lipopolysaccharide.  

OpenAIRE

Alkali-treated lipopolysaccharide from Brucella abortus S1119.3 coupled to agarose beads by cyanogen bromide activation resulted in an immunoadsorbent with which a large amount of B. abortus-specific antibodies could be purified. The method described gave alkali-treated lipopolysaccharide binding efficiencies of up to 98%. There was little loss of alkali-treated lipopolysaccharide from the column after several pH shifts, allowing the immunoadsorbent to be regenerated and used repeatedly.

Stiller, J. M.; Nielsen, K. H.

1983-01-01

308

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

Lifescience Database Archive (English)

Full Text Available 16357866 Lipopolysaccharide signaling in endothelial cells . Dauphinee SM, Karsan A. Lab Invest. ... ) Show Lipopolysaccharide signaling in endothelial cells . PubmedID 16357866 Title Lipopolysaccharide signal ... ing in endothelial cells . Authors Dauphinee SM, Karsan A. Publication Lab I ...

309

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

OpenAIRE

Since the pH of the gingival crevice increases from below neutrality in health to above pH 8 in disease, we decided to investigate the effect of environmental pH on the growth and enzyme activity of Bacteroides gingivalis W50. Cells were grown in a chemostat under hemin-excess conditions over a range of pH values; stable growth was observed only between pH 6.7 and 8.3, with the maximum yields obtained between pH 7.0 and 8.0. The enzyme profile of cells varied markedly with pH. Enzymes with a ...

Mcdermid, A. S.; Mckee, A. S.; Marsh, P. D.

1988-01-01

310

[Purification, characterization and induction by oxygen of superoxide dismutase from Bacteroides gingivalis 381].  

Science.gov (United States)

Several strains of Bacteroides gingivalis had strong activities of superoxide dismutase (SOD) and were markedly tolerant in the presence of oxygen in 13 strains of black-pigmented Bacteroides species tested. Thus, the strains were maintained and incubated in the either anaerobic or aerobic system. It was found that the SOD activity was significantly induced by oxygen, especially in B. gingivalis 381. The SODs, anaero-SOD and aero-SOD from the extracts of B. gingivalis 381 cells, each was purified by hydrophobic chromatography followed by anion exchange chromatography, and then by gel filtration, respectively. Both the purified enzymes having molecular weight of about 46,000 consisted of two subunits of equal sizes. Spectral analysis revealed that anaero-SOD had the characteristic A350 of Fe-SOD, but aero-SOD exhibited A475 of Mn-SOD. Both samples contained three isozymes with identical isoelectric points of 5.25, 5.10 and 5.00. On the basis of inactivation of SOD by H2O2, it was shown that aero-SOD consisted of one Mn-SOD and a small quantity of two Fe-SODs, whereas anaero-SOD contained only Fe-SOD. To ascertain whether or not the apoprotein of aero-SOD is the same as that of anaero-SOD, each apoprotein was prepared by dialysis in guanidinium chloride plus 8-hydroxyquinoline. Only one protein band with the same isoelectric point of 5.30 on an isoelectric focusing gel was obtained in each purified SOD sample. Subsequent reconstitution of both apoenzymes with either Fe (NH4)2 (SO4)2 or MnCl2 significantly restored their activity. These reconstituted SODs showed only one protein band with SOD activity on Native-PAGE. The complete amino acid sequence of anaero-SOD was determined by automated Edman degradation of the protein, Achrombacter protease I, endoproteinase Asp-N and tryptic digestion. The sequence consisted of 191 residues corresponding to a molecular weight of 21,500 per subunit. Furthermore, the first 36 amino acid sequence of aero-SOD was determined following N-terminal analysis. The two enzymes had similar amino acid compositions, and their amino-terminal sequences were identical through the first 36 amino acids in which methionine residue was present at N-terminal. These results suggest the three isozymes of either anaero-SOD or aero-SOD in B. gingivalis 381 may be formed from the same apoprotein. PMID:1966894

Amano, A

1990-12-01

311

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

312

[Phytotoxic properties of Ralstonia solanacearum lipopolysaccharides].  

Science.gov (United States)

The study is dedicated to research of phytotoxic properties of Ralstonia solanacearum lipopolysaccharides. This causative agent is one of the most dangerous among potato bacterial diseases. It is revealed that the inhibitory effect of LPS solution on seedlings germination is more noticeable on crops susceptible to brown rot. Maximal total phytotoxic properties have been shown by LPS from strains 35, 52b, TX1 and TS3, which were characterized by relatively low rhamnose content. Relative to the control plants LPS may diminish and some ones--increase the root length, height and weight of seedlings, subject to particular strain. But the stimulation revealed is minor. PMID:25000727

Hrytsa?, R V; Iakovleva, L M; Varbanets', L D

2014-01-01

313

A protease of Bacteroides gingivalis degrades cell surface and matrix glycoproteins of cultured gingival fibroblasts and induces secretion of collagenase and plasminogen activator.  

OpenAIRE

To assess the direct effects of Bacteroides gingivalis on periodontal cells, human gingival fibroblasts were cultured in the presence of B. gingivalis extracts or a trypsinlike enzyme partially purified from the bacteria by chromatography on benzamidine-Sepharose and Sephacryl S-200. Analysis of cell surface glycoproteins by the periodate-[3H]borohydride labeling technique combined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)-fluorography demonstrated that fibrone...

Uitto, V. J.; Larjava, H.; Heino, J.; Sorsa, T.

1989-01-01

314

Porphyromonas pogonae sp. nov., an anaerobic but low concentration oxygen adapted coccobacillus isolated from lizards (Pogona vitticeps) or human clinical specimens, and emended description of the genus Porphyromonas Shah and Collins 1988.  

Science.gov (United States)

During the process of identifying a Gram-negative coccobacillus isolated from a human clinical specimen, we found that the isolate's 16S rRNA gene had very close sequence identity with that of a variant Porphyromonas isolated from polymicrobial infections in the central bearded dragon, a species of lizard [2]. The 16S rRNA gene sequences of the human isolate and of six isolates from lizards were nearly identical (99.9-100%). Phylogenetic analysis placed all of these isolates in a single phylogenetic cluster well separated from other species in the genus Porphyromonas. The closest species was Porphyromonas catoniae with 90.7-90.9% sequence identity, although there was less than 6% DNA similarity between the P. catoniae type strain and our representative isolates from lizards (PAGU 1787(T)) and human (PAGU 1776). These isolates could grow under anaerobic or microaerobic conditions (6% O2 atmosphere). The isolates were positive for catalase and very strong ?-hemolytic activity, but did not show black or brown pigmentation. Biochemically, the isolates could be differentiated from closely related species by pyroglutamic acid arylamidase and glycine arylamidase activity, and some others. The fermentation products mainly included succinic acid and propionic acid. The major fatty acids detected in cells of the isolates were iso-C15:0, anteiso-C15:0, and 3OH-iso-C17:0. The G+C content was 43.0±0.62mol%. The species name Porphyromonas pogonae sp. nov. is proposed for these isolates with the type strain of PAGU 1787(T) (=MI 10-1288(T)=JCM 19732(T)=ATCC BAA-2643(T)). PMID:25481042

Kawamura, Yoshiaki; Kuwabara, Saki; Kania, Stephen A; Kato, Hisayuki; Hamagishi, Manami; Fujiwara, Nagatoshi; Sato, Takuichi; Tomida, Junko; Tanaka, Kaori; Bemis, David A

2015-03-01

315

[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

316

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

317

Lysozyme-mediated aggregation and lysis of the periodontal microorganism Capnocytophaga gingivalis 2010.  

Science.gov (United States)

The ability of lysozyme to aggregate and lyse the gram-negative capnophilic periodontal microorganism Capnocytophaga gingivalis 2010 was monitored optically at 540 nm. Both hen egg white and chromatographically purified human lysozymes had significant but similar aggregation potentials for both logarithmic- and stationary-phase bacteria. In general, an increase in enzyme concentration resulted in a graded increase in both the initial and maximum changes in turbidity which occurred during the reaction period. The greatest change in turbidity occurred within the initial minutes of interaction of lysozyme and the cells, and the extent of aggregation paralleled a rapid depletion of lysozyme by the suspensions during the first minute of its incubation with the bacteria. Interestingly, the muramidase inhibitors N-acetyl-D-glucosamine and histamine did not block aggregation, whereas maleylation of lysozyme completely inhibited its aggregating ability. Demaleylation, however, restored aggregation activity comparable to the native enzyme, indicating that maleylated lysozyme retained its integrity and that aggregation was primarily dependent on charge. The addition of up to physiological concentrations of NaHCO3 and NaCl to cell aggregates resulted in varying degrees of deaggregation and lysis. Surprisingly, ultrastructural analysis of lysozyme-treated cells revealed morphological changes with or without the addition of salt. Damage appeared to occur at the blunted polar end of the cells where there was a large spherical outpouching bordered by a damaged cell envelope. Damaged cells uniformly contained dense granular cytoplasmic debris. In effect, the cationic enzyme lysed C. gingivalis 2010, which was not apparent in the spectrophotometric assay. The paradoxical finding that during bacterial aggregation there was lysis may be of significance to the further elucidation of lysozyme's antibacterial role in the gingival sulcus. PMID:3967924

Iacono, V J; Zove, S M; Grossbard, B L; Pollock, J J; Fine, D H; Greene, L S

1985-02-01

318

Studies of lipopolysaccharides from Pseudomonas aeruginosa.  

Science.gov (United States)

Lipopolysaccharides from 13 strains of Pseudomonas aeruginosa representing seven serotypes of the Habs scheme have been analysed. The lipid A fractions, obtained by mild acid hydrolysis of the lipopolysaccharides, contained phosphorylated glucosamine residues substituted with dodecanoic, hexadecanoic, 2 hydroxydodecanoic, 3-hydroxydecanoic, and 3-hydroxydodecanoic acids (hexadecanoic acid and 2-hydroxdodecanoic acid were absent from one lipid A). Low-molecular-weight solutes released during the mild hydrolyses included 2-keto-3-deoxyoctonic acid, inorganic orthophosphates and pyrophosphates, ethanolamine mono, pyro and triphosphates. For most strains two polysaccharide fractions, one of which appeared to be the common core polysaccharide, were obtained. The major identifiable components and their approximate proportions in the core polysaccharides were glucose (3-4), rhamnose (1), galactosamine (1), alanine (1-1.5), phosphorus (4-6) and heptose (1-2). Rhamnose was absent from one polysaccharide another lacked both rhamnose and alanine but contained glucosamine. Small amounts of various amino sugars found in other core polysaccharides could be associated with the presence of higher-molecular-weight material. Such material was isolated from strain NCIB 8626. The high-molecular-weight polysaccharides obtained from ten strains were probably heterogeneous and consisted mainly of amino compounds, though rhamnose was a major component of four polysaccharades and arabinose was present in another. Fucosamine was the most common amino sugar, but quinovosamine, glucosamine, galactosamine, a possible aminohexuronic acid and unidentified amino compounds were also detected. The results of the survey are discussed in terms of the serological classification of the bacteria and of their sensitivity to EDTA. PMID:809266

Wilkinson, S G; Galbrath, L

1975-03-17

319

Lipopolysaccharide Extracellular Emulsifier Produced by Penicillium citrinum  

Directory of Open Access Journals (Sweden)

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

M.M. Camargo de Morais

2006-01-01

320

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.

2013-06-01

321

Lipopolysaccharide (LPS) stimulation of fungal secondary metabolism.  

Science.gov (United States)

We report on a preliminary investigation of the use the Gram-negative bacterial cell wall constituent lipopolysaccharide (LPS) as a natural chemical cue to stimulate and alter the expression of fungal secondary metabolism. Integrated high-throughput micro-cultivation and micro-analysis methods determined that 6 of 40 (15%) of fungi tested responded to an optimal exposure to LPS (0.6 ng/mL) by activating, enhancing or accelerating secondary metabolite production. To explore the possible mechanisms behind this effect, we employed light and fluorescent microscopy in conjunction with a nitric oxide (NO)-sensitive fluorescent dye and an NO scavenger to provide evidence that LPS stimulation of fungal secondary metabolism coincided with LPS activation of NO. Several case studies demonstrated that LPS stimulation can be scaled from single microplate well (1.5 mL) to preparative (>400 mL) scale cultures. For example, LPS treatment of Penicillium sp. (ACM-4616) enhanced pseurotin A and activated pseurotin A1 and pseurotin A2 biosynthesis, whereas LPS treatment of Aspergillus sp. (CMB-M81F) substantially accelerated and enhanced the biosynthesis of shornephine A and a series of biosynthetically related ardeemins and activated production of neoasterriquinone. As an indication of broader potential, we provide evidence that cultures of Penicillium sp. (CMB-TF0411), Aspergillus niger (ACM-4993F), Rhizopus oryzae (ACM-165F) and Thanatephorus cucumeris (ACM-194F) were responsive to LPS stimulation, the latter two examples being particular noteworthy as neither are known to produce secondary metabolites. Our results encourage the view that LPS stimulation can be used as a valuable tool to expand the molecular discovery potential of fungal strains that either have been exhaustively studied by or are unresponsive to traditional culture methodology. PMID:25379339

Khalil, Zeinab G; Kalansuriya, Pabasara; Capon, Robert J

2014-07-01

322

Porphyromonas gulae 41-kDa fimbriae induced osteoclast differentiation and cytokine production.  

Science.gov (United States)

Porphyromonas gulae is considered to be associated with canine periodontitis. We have previously reported that the P. gulae American Type Culture Collection (ATCC) 51700 comprised 41-kDa fimbriae. The purpose of the present study was to demonstrate the roles of 41-kDa fimbrial protein in periodontal disease. In this study, we examined the involvement of the 41-kDa fimbrial protein in osteoclast differentiation and cytokine production in murine macrophages. Furthermore, alveolar bone resorption induced by P. gulae infection in rats was evaluated. To estimate osteoclast differentiation, bone marrow cells and MC3T3-G2/PA6 cells were cultured with or without the 41-kDa fimbrial protein for 7 days. BALB/c mouse peritoneal macrophages were stimulated with the 41-kDa fimbrial protein, and the levels of interleukin (IL)-1? and tumor necrosis factor (TNF)-? production were determined by enzyme-linked immunosorbent assay. Osteoclast differentiation was significantly enhanced by treatment with the 41-kDa fimbrial protein in a dose-dependent manner. The total area of pits formed on the dentine slices with osteoclasts incubated with the 41-kDa fimbrial protein was significantly greater than that of the control. The purified 41-kDa fimbrial protein induced IL-1? and TNF-? production in BALB/c mouse peritoneal macrophages after 6 hr of incubation in a dose-dependent manner. The bone loss level in rats infected with P. gulae was significantly higher than that of the sham-infected rats. These results suggest that P. gulae 41-kDa fimbriae play important roles in the pathogenesis of periodontal disease. PMID:25421499

Sasaki, Haruka; Watanabe, Kiyoko; Toyama, Toshizo; Koyata, Yasunori; Hamada, Nobushiro

2014-11-25

323

Fosfomycin Alters Lipopolysaccharide-Induced Inflammatory Cytokine Production in Mice  

OpenAIRE

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

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

1999-01-01

324

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

Science.gov (United States)

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

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

2013-08-01

325

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

Scientific Electronic Library Online (English)

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 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

326

[Characterization of the lipopolysaccharides of serogroup II Azospirillum].  

Science.gov (United States)

Lipopolysaccharides of six Azospirillum strains (A. brasilense SR50, SR80, SR88, SR109, SR111, SR115, and A. lipoferum SR 42) isolated from the rhizosphere of cereal plants of Saratov oblast, Russia and assigned to serogroup II by serological analysis were studied. In the lipid A fatty acid composition, the lipopolysaccharides under study were similar to those of other Azospirillum strains and were characterized by predominance of 3-hydroxytetradecanoic, 3-hydroxyhexadecanoic, and octadecenoic acids. Monosaccharide analysis of the O-specific polysaccharides (including determination of the absolute configurations, methylation analysis, and one- and two-dimensional NMR spectroscopy) revealed the presence of two types of repeating units in varying ratios. High degree of serological similarity between the strains under study was shown to result from the presence of repeating units with identical structure in their O antigens. PMID:25844452

Sigida, E N; Fedonenko, Iu P; Zdorovenko, É L; Butygin, G L; Konnova, S A; Ignatov, V V

2014-01-01

327

Lipopolysaccharide induced inflammation in the perivascular space in lungs  

OpenAIRE

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. Meth...

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

2008-01-01

328

Compositional analysis of Helicobacter pylori rough-form lipopolysaccharides.  

OpenAIRE

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to analyze the macromolecular heterogeneity of lipopolysaccharides (LPS) from seven fresh clinical isolates and three culture collection strains of the human pathogen Helicobacter pylori. All the clinical isolates produced smooth-form LPS with O side chains of relatively homogeneous chain length, whereas the culture collection strains yielded rough-form LPS. A better yield of the latter LPS was obtained when combined protease ...

Moran, A. P.; Helander, I. M.; Kosunen, T. U.

1992-01-01

329

Biological activity of a lipopolysaccharide extracted from Treponema hyodysenteriae.  

OpenAIRE

A lipopolysaccharide (LPS) was obtained from pathogenic Treponema hyodysenteriae by hot phenol-water extraction. Various effects of the LPS on host cells were examined in vitro. Toxicity for mouse peritoneal macrophages was observed after 10 h of incubation at concentrations as low as 15 micrograms of the LPS per ml. Marked enhancement of both complement (C3) and immunoglobulin G-Fc receptor-mediated internalization was noted in macrophages obtained from mice injected 6 days previously with 7...

Nuessen, M. E.; Birmingham, J. R.; Joens, L. A.

1982-01-01

330

Impaired phagocyte responses to lipopolysaccharide in paroxysmal nocturnal hemoglobinuria.  

OpenAIRE

Bone marrow-derived cells from patients suffering from paroxysmal nocturnal hemoglobinuria (PNH) show a defect in the expression of phosphatidylinositol-anchored membrane proteins, including the CD14 molecule. Blocking experiments with anti-CD14 monoclonal antibodies have shown that lipopolysaccharide (LPS)-induced tumor necrosis factor alpha production by monocytes depends on the interaction between CD14 and a complex formed by LPS and LPS-binding protein. We used a whole-blood model to exam...

Duchow, J.; Marchant, A.; Crusiaux, A.; Husson, C.; Alonso-vega, C.; Groote, D.; Neve, P.; Goldman, M.

1993-01-01

331

Transcriptional Profiling of Lipopolysaccharide-Induced Acute Lung Injury  

OpenAIRE

Mortality associated with acute lung injury (ALI) induced by lipopolysaccharide (LPS) remains high in humans, warranting improved treatment and prevention strategies. ALI is characterized by the expression of proinflammatory mediators and extensive neutrophil influx into the lung, followed by severe lung damage. Understanding the pathogenesis of LPS-induced ALI is a prerequisite for designing better therapeutic strategies. In the present study, we used microarrays to gain a global view of the...

Jeyaseelan, Samithamby; Chu, Hong Wei; Young, Scott K.; Worthen, G. Scott

2004-01-01

332

Stimulation of peritoneal cell arginase by bacterial lipopolysaccharides.  

OpenAIRE

The conditions under which bacterial endotoxins stimulate arginase production in mouse peritoneal macrophages have been defined. Both lipid-A and lipid-A-associated protein are potent activators. Fetal calf serum and normal mouse serum enhance macrophage arginase levels in the presence and absence of lipopolysaccharide (LPS). LPS in the amount of 10(-1) microgram/ml represents a maximal stimulus for macrophage arginase production and release. Thioglycollate-elicited peritoneal cells have incr...

Ryan, J. L.; Yohe, W. B.; Morrison, D. C.

1980-01-01

333

Characterization of the Lipopolysaccharides and Capsules of Shewanella spp.  

OpenAIRE

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

334

Chemical and immunochemical analyses of Bacteroides fragilis lipopolysaccharides.  

OpenAIRE

Lipopolysaccharides (LPSs) from 17 different Bacteroides fragilis strains were extracted in a two-step procedure. The first step was a hot phenol-water extraction of whole bacteria, resulting in a crude aqueous phase, which after lyophilization in a second step was extracted with a phenol-chloroform-light petroleum mixture. The resulting LPSs, which were essentially free from contaminating nucleic acid, proteins, and capsular polysaccharide, were investigated for their qualitative and quantit...

Weintraub, A.; Larsson, B. E.; Lindberg, A. A.

1985-01-01

335

Induction of oral tolerance in mice unresponsive to bacterial lipopolysaccharide.  

OpenAIRE

C3H/HeJ lipopolysaccharide (LPS)-unresponsive and closely related C3H/HeSn LPS-responsive mice were rendered tolerant to hen albumin by antigen feeding before parenteral immunization. Both single and multiple feedings of antigen were effective in inducing tolerance to hen albumin in LPS-responsive and -unresponsive strains of mice. Our data demonstrate that ability to respond to LPS is not a prerequisite for induction of oral tolerance to a soluble protein antigen.

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

1983-01-01

336

Curcumin Attenuation of Lipopolysaccharide Induced Cardiac Hypertrophy in Rodents  

OpenAIRE

To study the ameliorating effects of curcumin in lipopolysaccharide (LPS) induced cardiac hypertrophy, mice were assigned to 4 groups (3 males and 3 females in each group): (A) control, (B) curcumin: 100??g/kg of body weight by intraperitoneal route (IP), (C) LPS: 60?mg/kg (IP), and (D) LPS + curcumin: both at previously stated concentrations by IP route. All mice were sacrificed as 12?hr and 24?hrs groups accordingly after LPS injection. The hearts were collected, photographed for c...

Rupak Chowdhury; Ramadevi Nimmanapalli; Thomas Graham; Gopal Reddy

2013-01-01

337

Comparison of Bacterial Lipopolysaccharides by High-Performance Liquid Chromatography  

OpenAIRE

A comparison of lipid-free polysaccharides from gram-negative bacteria was rapidly accomplished by using high-performance liquid chromatography of underivatized hydrolysates. Examination of a number of such products revealed that, contrary to earlier reports, Xanthomonas campestris lipopolysaccharide contained heptose, together with rhamnose and galactose, but not mannose. The polymers from the methanotrophs “Methylomonas albus” and “Methylosinus trichosporium” contained heptose and g...

Sutherland, Ian W.; Kennedy, Andrew F. D.

1986-01-01

338

Sero-characterization of lipopolysaccharide from Burkholderia thailandensis  

OpenAIRE

We report the successful purification of lipopolysaccharide (LPS) from Burkholderia thailandesis, a Gram-negative bacterium, closely related to the highly pathogenic organisms Burkholderia pseudomallei and Burkholderia mallei. B. thailandensis LPS is shown to cross-react with rabbit and mouse sera obtained from inoculation with B. pseudomallei or B. mallei, respectively. These data suggest that B. thailandensis LPS shares similar structural features with LPS molecules from highly pathogenic B...

Qazi, Omar; Prior, Joann L.; Judy, Barbara M.; Whitlock, Gregory C.; Kitto, G. Barrie; Torres, Alfredo G.; Estes, D. Mark; Brown, Katherine A.

2008-01-01

339

Chemical, biological, and immunochemical properties of the Chlamydia psittaci lipopolysaccharide.  

OpenAIRE

The lipopolysaccharide (LPS) of Chlamydia psittaci was extracted from yolk sac-grown elementary bodies, purified, and characterized chemically, immunochemically, and biologically. The LPS contained D-galactosamine, D-glucosamine, phosphorus, long-chain fatty acids, and 3-deoxy-D-manno-2-octulosonic acid in the molar ratio of approximately 1:2:2:6:5. The antigenic properties of the isolated LPS were compared with those of the LPS from Chlamydia trachomatis and Salmonella minnesota Re by the pa...

Brade, L.; Schramek, S.; Schade, U.; Brade, H.

1986-01-01

340

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

341

Lipopolysaccharide-Related Stimuli Induce Expression of the Secretory Leukocyte Protease Inhibitor, a Macrophage-Derived Lipopolysaccharide Inhibitor  

OpenAIRE

Mouse secretory leukocyte protease inhibitor (SLPI) was recently characterized as a lipopolysaccharide (LPS)-induced product of macrophages that antagonizes their LPS-induced activation of NF-?B and production of NO and tumor necrosis factor (TNF) (F. Y. Jin, C. Nathan, D. Radzioch, and A. Ding, Cell 88:417–426, 1997). To better understand the role of SLPI in innate immune and inflammatory responses, we examined the kinetics of SLPI expression in response to LPS, LPS-induced cytokines, and...

Jin, Fenyu; Nathan, Carl F.; Radzioch, Danuta; Ding, Aihao

1998-01-01

342

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

OpenAIRE

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.

2007-01-01

343

In vitro expression of tumor necrosis factor-?, interleukin 1?, and interleukin 8 mRNA by bovine macrophages following exposure to Porphyromonas levii  

OpenAIRE

The objective was to evaluate the pro-inflammatory response of bovine macrophages towards Porphyromonas levii, an etiologic agent of acute interdigital phlegmon, by evaluating the mRNA expression of tumor necrosis factor-? (TNF-?), interleukin 1? (IL-1?), and interleukin 8 (IL-8). Bovine macrophages detect the presence of bacteria, such as P. levii, and respond by upregulating transcription of the genes for pro-inflammatory cytokines TNF-? and IL-1? in addition to the neutrophil chemoat...

Walter, Michaela R. V.; Morck, Douglas W.

2002-01-01

344

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

Directory of Open Access Journals (Sweden)

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

Gabriella Di Francesco

2012-12-01

345

Interleukin-1 in Lipopolysaccharide Induced Chorioamnionitis in the Fetal Sheep  

OpenAIRE

We tested the hypothesis that interleukin 1 (IL-1) mediates intra-amniotic lipopolysaccharide (LPS)-induced chorioamnionitis in preterm fetal sheep. Time-mated Merino ewes with singleton fetuses received IL-1?, LPS, or saline (control) by intra-amniotic injection 1 to 2 days before operative delivery at 124 ± 1 days gestational age (N = 5-9/group; term = 150 days). Recombinant human IL-1 receptor antagonist (rhIL-1ra) was given into the amniotic fluid 3 hours before intra-amniotic LPS or sa...

Berry, Clare A.; Nitsos, Ilias; Hillman, Noah H.; Pillow, J. Jane; Polglase, Graeme R.; Kramer, Boris W.; Kemp, Matthew W.; Newnham, John P.; Jobe, Alan H.; Kallapur, Suhas G.

2011-01-01

346

Genetics and Proteomics of Aeromonas salmonicida Lipopolysaccharide Core Biosynthesis? †  

OpenAIRE

Comparison between the lipopolysaccharide (LPS) core structures of Aeromonas salmonicida subsp. salmonicida A450 and Aeromonas hydrophila AH-3 shows great similarity in the inner LPS core and part of the outer LPS core but some differences in the distal part of the outer LPS core (residues ld-Hep, d-Gal, and d-GalNAc). The three genomic regions encoding LPS core biosynthetic genes in A. salmonicida A450, of which regions 2 and 3 have genes identical to those of A. hydrophila AH-3, were fully ...

Jimenez, Natalia; Lacasta, Anna; Vilches, Silvia; Reyes, Merce?; Vazquez, Judit; Aquillini, Eleonora; Merino, Susana; Regue?, Miguel; Toma?s, Juan M.

2009-01-01

347

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

Lifescience Database Archive (English)

Full Text Available 15241548 Lipopolysaccharide-binding molecules: transporters, blockers and sensors . Chaby R. Cell ... ride-binding molecules: transporters, blockers and sensors . PubmedID 15241548 Title Lipopolysaccharide-bindin ... g molecules: transporters, blockers and sensors . Authors Chaby R. Publication Cell Mol Life Sci. 2 ...

348

Distribution of porin and lipopolysaccharide antigens on a Pseudomonas aeruginosa permeability mutant.  

OpenAIRE

Pseudomonas aeruginosa common surface antigens were compared in a permeability mutant (PCC118) and its parent (PAO503). The distribution of lipopolysaccharide and porin antigens in the mutant supports the conclusion that beta-lactam permeability was affected by lipopolysaccharide-side chain presentation rather than by a change in porin number.

Godfrey, A. J.; Shahrabadi, M. S.; Bryan, L. E.

1986-01-01

349

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

Lifescience Database Archive (English)

Full Text Available 15051069 Innate recognition of lipopolysaccharide ... by Toll-like receptor 4-MD-2. Miyake K. Trends ... (.svg) (.html) (.csml) Show Innate recognition of lipopolysaccharide ... by Toll-like receptor 4-MD-2. PubmedID 15051069 Ti ... tle Innate recognition of lipopolysaccharide ... by Toll-like receptor 4-MD-2. Authors Miyake K. Pu ...

350

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

Lifescience Database Archive (English)

Full Text Available 15379975 Signal transduction by the lipopolysaccharide ... receptor, Toll-like receptor-4. Palsson-M ... g) (.html) (.csml) Show Signal transduction by the lipopolysaccharide ... receptor, Toll-like receptor-4. PubmedID 15379975 ... Title Signal transduction by the lipopolysaccharide ... receptor, Toll-like receptor-4. Authors Palsson-Mc ...

351

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 (Pdiets. Whereas pigs fed diet WB had about 2 to 22 times higher abundance (Pdiets. The total amount of digested NSP (r=0.57; P=0.002), xylose (r=0.53; P=0.004) and dietary fiber (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

352

Bovine recombinant lipopolysaccharide binding protein (BRLBP) regulated apoptosis and inflammation response in lipopolysaccharide-challenged bovine mammary epithelial cells (BMEC).  

Science.gov (United States)

Lipopolysaccharide-binding protein (LBP) is an acute-phase protein involved in host response to Gram-negative and Gram-positive pathogens. It has been reported to exert diverse biological activities, such as anti-inflammatory effects. However, what effects it has on bovine mastitis has not been investigated. The aim of this study was to verify the anti-inflammatory properties of LBP on the inflammatory response of primary bovine mammary epithelial cells (BMEC) induced by lipopolysaccharide (LPS), and to determine the underlying mechanism. Bovine mammary epithelial cells were treated with various concentrations of LPS (1, 10, 20, and 100?g/mL) for 3, 6, 12, and 24h. The results showed that LPS significantly inhibited cell viability in a dose-dependent manner. When cells were treated with LPS (10?g/mL) for 12h, the permeability of the cell membrane increased significantly. This promoted apoptosis. Various concentrations (10 and 20?g/mL) of bovine recombinant lipopolysaccharide binding protein (BRLBP) could weaken the inflammation injury of BMEC induced by LPS without cytotoxicity. Toll-like receptor 4 (TLR4), nuclear factor ?B (NF-?B), IL-1?, and tumor necrosis factor ? (TNF-?) from BMEC were decreased. TLR4 and NF-?B P65 protein levels were down-regulated, and nuclear transcription factor ?B activity was also weakened. All these results indicated that the protective effects of high concentrations of BRLBP on LPS-induced inflammation injury in BMEC were at least partially achieved by the decreased production of pro-inflammatory cytokines. BRLBP was found to directly inhibit LPS/TLR4-mediated NF-?B activation. One possible anti-inflammatory mechanism can be attributed to the negative role of BRLBP in suppressing TLR4/NF-?B activation mediated by LPS. These findings suggested that BRLBP may be a useful agent to treat LPS-induced mastitis. PMID:25700343

Sun, Yu; Li, Lian; Wu, Jie; Yu, Pan; Li, Chengmin; Tang, Juan; Li, Xiaojuan; Huang, Shuai; Wang, Genlin

2015-06-01

353

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

354

Recognition of lipopolysaccharide pattern by TLR4 complexes  

Science.gov (United States)

Lipopolysaccharide (LPS) is a major component of the outer membrane of Gram-negative bacteria. Minute amounts of LPS released from infecting pathogens can initiate potent innate immune responses that prime the immune system against further infection. However, when the LPS response is not properly controlled it can lead to fatal septic shock syndrome. The common structural pattern of LPS in diverse bacterial species is recognized by a cascade of LPS receptors and accessory proteins, LPS binding protein (LBP), CD14 and the Toll-like receptor4 (TLR4)–MD-2 complex. The structures of these proteins account for how our immune system differentiates LPS molecules from structurally similar host molecules. They also provide insights useful for discovery of anti-sepsis drugs. In this review, we summarize these structures and describe the structural basis of LPS recognition by LPS receptors and accessory proteins. PMID:24310172

Park, Beom Seok; Lee, Jie-Oh

2013-01-01

355

Lipopolysaccharide induces amyloid formation of antimicrobial peptide HAL-2.  

Science.gov (United States)

Lipopolysaccharide (LPS), the important component of the outer membrane of Gram-negative bacteria, contributes to the integrity of the outer membrane and protects the cell against bactericidal agents, including antimicrobial peptides. However, the mechanisms of interaction between antimicrobial peptides and LPS are not clearly understood. Halictines-2 (HAL-2), one of the novel antimicrobial peptides, was isolated from the venom of the eusocial bee Halictus sexcinctus. HAL-2 has exhibited potent antimicrobial activity against Gram-positive and Gram-negative bacteria and even against cancer cells. Here, we studied the interactions between HAL-2 and LPS to elucidate the antibacterial mechanism of HAL-2 in vitro. Our results show that HAL-2 adopts a significant degree of ?-strand structure in the presence of LPS. LPS is capable of inducing HAL-2 amyloid formation, which may play a vital role in its antimicrobial activity. PMID:25109934

Wang, Jiarong; Li, Yan; Wang, Xiaoming; Chen, Wei; Sun, Hongbin; Wang, Junfeng

2014-11-01

356

Immune modulation of Prevotella intermedia colonization in squirrel monkeys.  

OpenAIRE

Colonization of the gingival crevice by black-pigmented Porphyromonas or Prevotella spp. (BP/P), including Porphyromonas gingivalis (formerly Bacteroides gingivalis) and Prevotella intermedia (formerly Bacteroides intermedius), is thought to be an important ecological event which may result in the destruction of connective tissues supporting the teeth. Theoretically, periodontal diseases could be prevented if these or other periodontal pathogenic microorganisms did not colonize the subgingiva...

Clark, W. B.; Magnusson, I.; Beem, J. E.; Jung, J. M.; Marks, R. G.; Mcarthur, W. P.

1991-01-01

357

Interactions between lipopolysaccharide and outer membrane proteins of Acinetobacter calcoaceticus studied by an affinity electrophoresis system.  

Science.gov (United States)

R-Form lipopolysaccharides of Acinetobacter calcoaceticus could be incorporated into polyacrylamide gels in an immobile form by adding it directly to the acrylamide-N,N'-methylenebisacrylamide polymerization mixture. The separation of A. calcoaceticus 69 V outer membrane proteins in these affinity gels demonstrated a specific interaction with the lipopolysaccharide ligand for one of the proteins. This protein is heat-modifiable and has an Mr of about 18,000. By incorporation of varying concentrations of lipopolysaccharide, a dissociation constant of the protein-lipopolysaccharide complex of 0.5 mM could be determined. In comparison, for another A. calcoaceticus strain, CCM 5593, a higher dissociation constant (1.0 mM)--indicative of lower affinity--was obtained. PMID:2743966

Borneleit, P; Blechschmidt, B; Kleber, H P

1989-04-01

358

Acanthoic Acid inhibits LPS-induced inflammatory response in human gingival fibroblasts.  

Science.gov (United States)

Periodontitis is a chronic disease that affects the gums and destroys connective tissue. Acanthoic acid (AA), a diterpene in Acanthopanax koreanum, has been reported to have anti-inflammatory activities. The aim of this study was to investigate the anti-inflammatory effects of AA on lipopolysaccharide (LPS)-induced inflammatory response in human gingival fibroblasts (HGFs). HGFs were treated with Porphyromonas gingivalis LPS in the presence or absence of AA. The production of inflammatory cytokines IL-8 and IL-6 were measured by ELISA. The expression of NF-?B and TLR4 were detected by Western blotting. The results showed that AA inhibited LPS-induced IL-8 and IL-6 production in a dose-dependent manner. In addition, AA inhibited LPS-induced TLR4 expression and NF-?B activation. In conclusion, AA inhibits LPS-induced inflammatory response in HGFs through inhibition TLR4-mediated NF-?B signaling pathway. PMID:25373915

Wei, Cai; Tan, Chee Keong; Xiaoping, He; Junqiang, Jiang

2015-04-01

359

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

Science.gov (United States)

Xanthomonas axonopodis pv. citri (Xac) causes citrus canker, provoking defoliation and premature fruit drop with concomitant economical damage. In plant pathogenic bacteria, lipopolysaccharides are important virulence factors, and they are being increasingly recognized as major pathogen-associated molecular patterns for plants. In general, three domains are recognized in a lipopolysaccharide: the hydrophobic lipid A, the hydrophilic O-antigen polysaccharide, and the core oligosaccharide, connecting lipid A and O-antigen. In this work, we have determined the structure of purified lipopolysaccharides obtained from Xanthomonas axonopodis pv. citri wild type and a mutant of the O-antigen ABC transporter encoded by the wzt gene. High pH anion exchange chromatography and matrix-assisted laser desorption/ionization mass spectrum analysis were performed, enabling determination of the structure not only of the released oligosaccharides and lipid A moieties but also the intact lipopolysaccharides. The results demonstrate that Xac wild type and Xacwzt LPSs are composed mainly of a penta- or tetra-acylated diglucosamine backbone attached to either two pyrophosphorylethanolamine groups or to one pyrophosphorylethanolamine group and one phosphorylethanolamine group. The core region consists of a branched oligosaccharide formed by Kdo?Hex?GalA?Fuc3NAcRha? and two phosphate groups. As expected, the presence of a rhamnose homo-oligosaccharide as O-antigen was determined only in the Xac wild type lipopolysaccharide. In addition, we have examined how lipopolysaccharides from Xac function in the pathogenesis process. We analyzed the response of the different lipopolysaccharides during the stomata aperture closure cycle, the callose deposition, the expression of defense-related genes, and reactive oxygen species production in citrus leaves, suggesting a functional role of the O-antigen from Xac lipopolysaccharides in the basal response. PMID:21596742

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

2011-07-22

360

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

OpenAIRE

Xanthomonas axonopodis pv. citri (Xac) causes citrus canker, provoking defoliation and premature fruit drop with concomitant economical damage. In plant pathogenic bacteria, lipopolysaccharides are important virulence factors, and they are being increasingly recognized as major pathogen-associated molecular patterns for plants. In general, three domains are recognized in a lipopolysaccharide: the hydrophobic lipid A, the hydrophilic O-antigen polysaccharide, and the core oligosaccharide, conn...

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

2011-01-01

361

Lipopolysaccharide Desensitization of Macrophages Provides Protection against Yersinia enterocolitica-Induced Apoptosis  

OpenAIRE

Pathogenic Yersinia spp. uncouple an array of signal transduction pathways in macrophages to disrupt their response to infection. This compels the macrophage to undergo apoptosis. Our study shows that macrophages that had acquired tolerance to Yersinia infection by preexposure to lipopolysaccharide were considerably protected against Y. enterocolitica-induced apoptosis. The desensitization of macrophages by lipopolysaccharide, which is thought to be a self-protective, adaptive response to sus...

Ruckdeschel, Klaus; Richter, Kathleen

2002-01-01

362

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

363

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

International Nuclear Information System (INIS)

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

364

Lipopolysaccharide induces autotaxin expression in human monocytic THP-1 cells  

International Nuclear Information System (INIS)

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

365

Arctigenin attenuates lipopolysaccharide-induced acute lung injury in rats.  

Science.gov (United States)

Arctigenin (ATG) has been reported to possess anti-inflammatory properties. However, the effects of ATG on lipopolysaccharide (LPS)-induced acute lung injury (ALI) remains not well understood. In the present study, our investigation was designed to reveal the effect of ATG on LPS-induced ALI in rats. We found that ATG pretreatment attenuated the LPS-induced ALI, as evidenced by the reduced histological scores, myeloperoxidase activity, and wet-to-dry weight ratio in the lung tissues. This was accompanied by the decreased levels of tumor necrosis factor alpha (TNF-?), interleukin-1? (IL-1?), and interleukin-1 (IL-6) in the bronchoalveolar lavage fluid. Furthermore, ATG downregulated the expression of nuclear factor kappa B (NF-?B) p65, promoted the phosphorylation of inhibitor of nuclear factor-?B-? (I?B?) and activated the adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK?) in the lung tissues. Our results suggested that ATG attenuates the LPS-induced ALI via activation of AMPK and suppression of NF-?B signaling pathway. PMID:25008149

Shi, Xianbao; Sun, Hongzhi; Zhou, Dun; Xi, Huanjiu; Shan, Lina

2015-04-01

366

Induction of glomerular alkaline phosphatase after challenge with lipopolysaccharide  

Science.gov (United States)

Alkaline phosphatase (AP) can be considered as a host defence molecule since this enzyme is able to detoxify bacterial endotoxin at physiological pH. The question emerged whether this anti-endotoxin principle is inducible in the glomerulus and if so, which glomerular cells might be involved in the expression of ectoAP after stimulation with pro-inflammatory agents. Therefore kidneys of rats treated with either lipopolysaccharide (LPS), E. coli bacteria or non-toxic monophosphoryl lipid A (MPLA) were examined for AP activity 6 or 24 h after challenge. In addition cultures of endothelial cells or mesangial cells were evaluated for AP activity after stimulation with either LPS, TNF? or IL-6, and mRNA for AP was studied in TNF?-stimulated and control mesangial cells. The results show significant up-regulation of glomerular AP in LPS- or E. coli-injected rats compared to rats injected with MPLA. Endothelial and mesangial cells in vitro showed significant up-regulation of AP activity following stimulation with LPS, TNF? or IL-6, whereas increased mRNA for AP was observed in mesangial cells after TNF? stimulation compared to non-stimulated control cells. Since it appeared that hydrolysis occurred when endotoxin was used as a substrate in the histochemical staining, we concluded that inducible glomerular ectoAP may reflect a local endotoxin detoxifying principle of the kidney. PMID:12974943

Kapojos, Jola J; Poelstra, Klaas; Borghuis, Theo; Van Den Berg, Anke; Baelde, Hans J; Klok, Pieter A; Bakker, Winston W

2003-01-01

367

Lipopolysaccharide-induced inflammatory liver injury in mice.  

Science.gov (United States)

The intraperitoneal application of lipopolysaccharide (LPS) alone or in combination with other hepatotoxins is an experimental model for inducing systemic and hepatic inflammation in rodents applied worldwide. The endotoxin is recognized by the LPS-binding protein. This complex binds together with the lymphocyte antigen 96 (MD2) and the pattern-recognition receptor CD14 to members of the toll-like receptor family. The activated receptor complex in turn transduces signals to well characterized intracellular cascades that result in a multifaceted network of intracellular responses ending in inflammation. The most prominent among these is the activation of the NF-?B pathway and the production of a multitude of inflammatory cytokines. Although the application of LPS is in general easy to perform, unintended variations in preparation of the injection solution or in handling of the animals might affect the reproducibility or the outcome of a specific experiment. Here, we present a well-standardized protocol that allows for an induction of highly reproducible acute hepatic inflammation in mice. Furthermore, examples of appropriate readouts for the resulting inflammatory response are given. PMID:25835737

Hamesch, K; Borkham-Kamphorst, E; Strnad, P; Weiskirchen, R

2015-04-01

368

Roles of different forms of lipopolysaccharides in Ralstonia solanacearum pathogenesis.  

Science.gov (United States)

Lipopolysaccharides (LPS) are critical components for the fitness of most gram-negative bacteria. Ralstonia solanacearum causes a deadly wilting disease in many crops; however, the pathogenic roles of different forms of LPS and their pathways of biogenesis remain unknown. By screening for phage-resistant mutants of R. solanacearum Pss4, whose genome sequence is unavailable, mutants with various types of structural defects in LPS were isolated. Pathogenesis assays of the mutants revealed that production of rough LPS (R-LPS), which does not contain O-polysaccharides, was sufficient to cause necrosis on Nicotiana benthamiana and induce the hypersensitive response on N. tabacum. However, biosynthesis of smooth LPS (S-LPS), which contains O-polysaccharides, was required for bacterial proliferation at infection sites on N. benthamiana leaves and for proliferation and causing wilt on tomato. Complementation tests confirmed the involvement of the previously unidentified cluster RSc2201 to RSc2204 in the formation of R. solanacearum S-LPS. With these data and the availability of the annotated genomic sequence of strain GMI1000, certain loci involved in key steps of R. solanacearum LPS biosynthesis were identified. The strategy of this work could be useful for similar studies in other bacteria without available genome sequences. PMID:24580105

Li, Chien-Hui; Wang, Kuan-Chung; Hong, Yu-Hau; Chu, Tai-Hsiang; Chu, Yu-Ju; Chou, I-Chun; Lu, Der-Kang; Chen, Chiao-Yen; Yang, Wen-Chieh; Lin, Yu-Mei; Cheng, Chiu-Ping

2014-05-01

369

Lipopolysaccharide exposure augments isoniazide-induced liver injury.  

Science.gov (United States)

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. PMID:25331106

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

2014-12-01

370

Lipopolysaccharide aggravates bisphosphonate-induced osteonecrosis in rats.  

Science.gov (United States)

The pathogenesis of bisphosphonate-related osteonecrosis of the jaw (BRONJ) is highly controversial. We have previously reported the development of osteonecrosis by periodontal pathogenic stimulation in the jaw and femur of rats treated with bisphosphonate. Since the major toxicity factor of Gram-negative bacteria is lipopolysaccharide (LPS), the present study aimed to evaluate the relationship between osteonecrosis and LPS in a rat model of BRON-like lesions. Seventeen male rats were injected subcutaneously with zoledronic acid weekly for 4 weeks and divided into three groups: LPS (LPS administered into the bone marrow of the mandible and femur) and LPS plus polymyxin B (PMB) and saline groups (given neutralized LPS with PMB or saline, respectively, using the same protocol). At 4 weeks after the procedure, harvested specimens were analyzed using histomorphology (n=5 from each group) and histochemistry (n=1 each from LPS and LPS plus PMB groups). There was a significantly wider area of osteonecrosis in the LPS group as compared to the saline and LPS plus PMB groups in both the mandible (P=0.030 and P=0.009, respectively) and femur (P=0.002 and P=0.020, respectively). Our results indicate that LPS stimulation is deeply involved in the development and promotion of BRON. PMID:25442743

Sakaguchi, O; Kokuryo, S; Tsurushima, H; Tanaka, J; Habu, M; Uehara, M; Nishihara, T; Tominaga, K

2015-04-01

371

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

372

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

373

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

374

RNA interference prevents lipopolysaccharide-induced preprotachykinin gene expression  

International Nuclear Information System (INIS)

We showed previously that lipopolysaccharide (LPS) induces noncholinergic airway hyperreactivity to capsaicin via an upregulation of tachykinin synthesis. This study was designed to test whether double-stranded preprotachykinin (ds PPT) RNA, RNA interference (RNAi), prevents the LPS-induced alterations. First, cultured primary nodose ganglial cells of newborn Brown-Norway rats were divided into four groups: control; LPS; LPS+RNAi; and LPS+RNAi+liposome. Second, young Brown-Norway rats for the in vivo study were divided into three groups (control; LPS; and LPS+RNAi), and ds PPT RNA was microinjected bilaterally into the nodose ganglia in the LPS+RNAi group. Then, ganglial cells were collected from the culture whereas the nodose ganglia and lungs were sampled from the animals, and PPT mRNA and substance P (SP) levels were analyzed. Also, airway reactivity to capsaicin was performed in vivo. LPS induced significant increases in PPT mRNA and SP levels in vitro and in vivo and an increase in airway reactivity to capsaicin in vivo. However, ds PPT RNA, but not scrambled RNA, prevented all LPS-induced alterations. The effect of ds PPT RNA was not enhanced by liposome in vitro. Therefore, we demonstrated that the local application of RNAi prevents effectively the activation of the noncholinergic system modulating the lungs/airways

375

Lipopolysaccharide induced inflammation in the perivascular space in lungs  

Directory of Open Access Journals (Sweden)

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

Pabst Reinhard

2008-07-01

376

Bitter gourd suppresses lipopolysaccharide-induced inflammatory responses.  

Science.gov (United States)

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

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

2008-06-11

377

Melatonin protects against myocardial hypertrophy induced by lipopolysaccharide.  

Science.gov (United States)

Melatonin is thought to have the ability of antiatherogenic, antioxidant, and vasodilatory. It is not only a promising protective in acute myocardial infarction but is also a useful tool in the treatment of pathological remodeling. However, its role in myocardial hypertrophy remains unclear. In this study, we investigated the protective effects of melatonin on myocardial hypertrophy induced by lipopolysaccharide (LPS) and to identify their precise mechanisms. The cultured myocardial cell was divided into six groups: control group, LPS group, LPS + ethanol (4%), LPS + melatonin (1.5 mg/ml) group, LPS + melatonin (3 mg/ml) group, and LPS + melatonin (6 mg/ml) group. The morphologic change of myocardial cell was observed by inverted phase contrast microscope. The protein level of myocardial cell was measured by Coomassie brilliant blue protein kit. The secretion level of tumor necrosis factor-? (TNF-?) was evaluated by enzyme-linked immunosorbent assay (ELISA). Ca(2+) transient in Fura-2/AM-loaded cells was measured by Till image system. The expression of Ca(2+)/calmodulin-dependent kinase II (CaMKII) and calcineurin (CaN) was measured by Western blot analysis. Our data demonstrated that LPS induced myocardial hypertrophy, promoted the secretion levels of TNF-?, and increased Ca(2+) transient level and the expression of CaMKII and CaN. Administration of melatonin 30 min prior to LPS stimulation dose-dependently attenuated myocardial hypertrophy. In conclusion, the results revealed that melatonin had the potential to protect against myocardial hypertrophy induced by LPS in vitro through downregulation of the TNF-? expression and retains the intracellular Ca(2+) homeostasis. PMID:25475041

Lu, Qi; Yi, Xin; Cheng, Xiang; Sun, Xiaohui; Yang, Xiangjun

2015-04-01

378

The nature of lipopolysaccharide-stimulated monocyte tissue factor activity.  

Science.gov (United States)

There are several contradictory hypotheses that attempt to explain changes in cell tissue factor (TF) activity upon treatment with various agents ('encryption-decryption'). We evaluated the influence of lipopolysaccharide stimulation on expression of TF antigen and activity on/in THP-1 human leukemia monocytic cells. Prior to stimulation, there were 240?±?60 TF molecules/cell on the cell surface and 510?±?180?molecules/cell in lysates (n?=?8). Upon stimulation, TF antigen increased 10-fold on the cell surface and 16.5-fold in lysates. Coincidently, the intact cell factor (F)Xa generation by TF/FVIIa increased 11-fold. Correspondingly, TF-induced clotting activity increased 35.7?±?4.9-fold. The KM of the TF/FVIIa complex formed on the THP-1 surface and cell lysates for FX was 0.73?±?0.07 and 0.41?±?0.02??mol/l and the kcat 59.4?±?1.8 and 44.6?±?0.1?s, respectively. For isolated and relipidated THP-1 cell TF, the efficiency of FXase was lower (KM?=?1.54??mol/l and kcat?=?12.0?s). A similar comparison of isolated/relipidated vs. cell-surface TF in the synthetic coagulation proteome and corn trypsin inhibitor blood showed that the cell-surface TF-induced thrombin generation is more than 100-fold more efficient than that induced by purified/relipidated TF. These data indicate that the increase in monocytic cell TF activity upon stimulation is primarily related to an increased expression of TF protein, and that significantly higher specific activity of TF presented on cells than that of purified and relipidated protein suggests the presence of the cell membrane components which substantially enhance TF function. PMID:24047888

Butenas, Saulius; Gissel, Matthew; Krudysz-Amblo, Jolanta; Mann, Kenneth G

2013-09-16

379

Bacterial lipopolysaccharide promotes profibrotic activation of intestinal fibroblasts.  

LENUS (Irish Health Repository)

BACKGROUND: Fibroblasts play a critical role in intestinal wound healing. Lipopolysaccharide (LPS) is a cell wall component of commensal gut bacteria. The effects of LPS on intestinal fibroblast activation were characterized. METHODS: Expression of the LPS receptor, toll-like receptor (TLR) 4, was assessed in cultured primary human intestinal fibroblasts using flow cytometry and confocal microscopy. Fibroblasts were treated with LPS and\\/or transforming growth factor (TGF) beta1. Nuclear factor kappaB (NFkappaB) pathway activation was assessed by inhibitory kappaBalpha (IkappaBalpha) degradation and NFkappaB promoter activity. Fibroblast contractility was measured using a fibroblast-populated collagen lattice. Smad-7, a negative regulator of TGF-beta1 signalling, and connective tissue growth factor (CTGF) expression were assessed using reverse transcriptase-polymerase chain reaction and western blot. The NFkappaB pathway was inhibited by IkappaBalpha transfection. RESULTS: TLR-4 was present on the surface of intestinal fibroblasts. LPS treatment of fibroblasts induced IkappaBalpha degradation, enhanced NFkappaB promoter activity and increased collagen contraction. Pretreatment with LPS (before TGF-beta1) significantly increased CTGF production relative to treatment with TGF-beta1 alone. LPS reduced whereas TGF-beta1 increased smad-7 expression. Transfection with an IkappaBalpha plasmid enhanced basal smad-7 expression. CONCLUSION: Intestinal fibroblasts express TLR-4 and respond to LPS by activating NFkappaB and inducing collagen contraction. LPS acts in concert with TGF-beta1 to induce CTGF. LPS reduces the expression of the TGF-beta1 inhibitor, smad-7.

Burke, J P

2012-02-01

380

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 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

381

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

382

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

383

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

Lifescience Database Archive (English)

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

384

Maturational effects of lipopolysaccharide on white-matter injury in fetal sheep.  

Science.gov (United States)

White-matter damage has been associated with the development of cerebral palsy in children born both prematurely and at term, and it has been suggested that intrauterine infection can contribute to the brain injury. However, the relative importance of age on white-matter injury following infectious exposure in utero remains unclear. In this study, fetal sheep were exposed to systemic endotoxemia by administration of Escherichia coli lipopolysaccharide (88.7 +/- 7.7 ng/kg) at 65% or 85% of gestation. These gestational ages approximately correspond to human brain development in preterm and near-term infants respectively. White-matter injury was evaluated 3 days after lipopolysaccharide exposure with regard to microglia activation and loss of neurofilament and myelin basic protein. The expression of oligodendrocytes at different maturational stages was demonstrated in preterm and near-term fetuses with the oligodendroglial markers O4 and 2 ,3 -cyclic nucleotide 3 -phospodiesterase. Forty percent of the fetuses in the preterm group and 22% in the near-term group died within 8 hours of the endotoxin exposure. Three of six preterm and two of seven near-term surviving fetuses demonstrated pathologic changes in the brain with regard to increased microglia activation and loss of neurofilament staining. The number of activated microglia was enhanced in the subcortical white matter in both the preterm lipopolysaccharide-exposed fetuses (lipopolysaccharide: 235 +/- 64 cells/mm2; control: 72 +/- 28 cells/mm2; P = .0374) and the near-term fetuses (lipopolysaccharide: 180 +/- 40 cells/mm2; control 23 +/- 16 cells/mm2; P = .0152). There was a loss of neurofilament staining in both preterm fetuses (lipopolysaccharide: 2.20 +/- 0.77 pixel units; control: 0.20 +/- 0.10 pixel units; P = .0306) and near-term fetuses (lipopolysaccharide: 1.15 +/- 0.48 pixel units; control: 0.06 +/- 0.06 pixel units; P = .0285). O4-positive cells were detected at both gestational ages, whereas 2,3-cyclic nucleotide 3-phospodiesterase-positive cells and myelin basic protein staining were mainly detected in the near-term fetuses. In summary, we found white-matter injury in a proportion of both preterm and near-term fetuses after administration of lipopolysaccharide. These results are in agreement with clinical evidence suggesting that both preterm and term infants are at risk of periventricular leukomalacia in association with intrauterine infection. PMID:16417842

Svedin, Pernilla; Kjellmer, Ingmar; Welin, Anna-Karin; Blad, Sofia; Mallard, Carina

2005-12-01

385

Lipid lateral organization on giant unilamellar vesicles containing lipopolysaccharides  

DEFF Research Database (Denmark)

We developed a new (to our knowledge) protocol to generate giant unilamellar vesicles (GUVs) composed of mixtures of single lipopolysaccharide (LPS) species and Escherichia coli polar lipid extracts. Four different LPSs that differed in the size of the polar headgroup (i.e., LPS smooth > LPS-Ra > LPS-Rc > LPS-Rd) were selected to generate GUVs composed of different LPS/E. coli polar lipid mixtures. Our procedure consists of two main steps: 1), generation and purification of oligolamellar liposomes containing LPSs; and 2), electroformation of GUVs using the LPS-containing oligolamellar vesicles at physiological salt and pH conditions. Analysis of LPS incorporation into the membrane models (both oligolamellar vesicles and GUVs) shows that the final concentration of LPS is lower than that expected from the initial E. coli lipids/LPS mixture. In particular, our protocol allows incorporation of no more than 15 mol % for LPS-smooth and LPS-Ra, and up to 25 mol % for LPS-Rc and LPS-Rd (with respect to total lipids).We used the GUVs to evaluate the impact of different LPS species on the lateral structure of the host membrane (i.e., E. coli polar lipid extract). Rhodamine-DPPE-labeled GUVs show the presence of elongated micrometer-sized lipid domains for GUVs containing either LPS-Rc or LPS-Rd above 10 mol %. Laurdan GP images confirm this finding and show that this particular lateral scenario corresponds to the coexistence of fluid disordered and gel (LPS-enriched)-like micron-sized domains, in similarity to what is observed when LPS is replaced with lipid A. For LPSs containing the more bulky polar headgroup (i.e., LPS-smooth and LPS-Ra), an absence of micrometer-sized domains is observed for all LPS concentrations explored in the GUVs (up to ?15 mol %). However, fluorescence correlation spectroscopy (using fluorescently labeled LPS) and Laurdan GP experiments in these microscopically homogeneous membranes suggests the presence of LPS clusters with dimensions below our microscope's resolution (?380 nm radial). Our results indicate that LPSs can cluster into gel-like domains in these bacterial model membranes, and that the size of these domains depends on the chemical structure and concentration of the LPSs.

Kubiak, Jakub; Brewer, Jonathan R.

2011-01-01

386

Nilotinib ameliorates lipopolysaccharide-induced acute lung injury in rats  

International Nuclear Information System (INIS)

The present study aimed to investigate the effect of the new tyrosine kinase inhibitor, nilotinib on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in rats and explore its possible mechanisms. Male Sprague-Dawley rats were given nilotinib (10 mg/kg) by oral gavage twice daily for 1 week prior to exposure to aerosolized LPS. At 24 h after LPS exposure, bronchoalveolar lavage fluid (BALF) samples and lung tissue were collected. The lung wet/dry weight (W/D) ratio, protein level and the number of inflammatory cells in the BALF were determined. Optical microscopy was performed to examine the pathological changes in lungs. Malondialdehyde (MDA) content, superoxidase dismutase (SOD) and reduced glutathione (GSH) activities as well as nitrite/nitrate (NO2-/NO3-) levels were measured in lung tissues. The expression of inflammatory cytokines, tumor necrosis factor-? (TNF-?), transforming growth factor-?1 (TGF-?1) and inducible nitric oxide synthase (iNOS) were determined in lung tissues. Treatment with nilotinib prior to LPS exposure significantly attenuated the LPS-induced pulmonary edema, as it significantly decreased lung W/D ratio, protein concentration and the accumulation of the inflammatory cells in the BALF. This was supported by the histopathological examination which revealed marked attenuation of LPS-induced ALI in nilotinib treated rats. In addition, nilotinib significantly increased SO, nilotinib significantly increased SOD and GSH activities with significant decrease in MDA content in the lung. Nilotinib also reduced LPS mediated overproduction of pulmonary NO2-/NO3- levels. Importantly, nilotinib caused down-regulation of the inflammatory cytokines TNF-?, TGF-?1 and iNOS levels in the lung. Taken together, these results demonstrate the protective effects of nilotinib against the LPS-induced ALI. This effect can be attributed to nilotinib ability to counteract the inflammatory cells infiltration and hence ROS generation and regulate cytokine effects. - Research highlights: ? The protective effects of nilotinib against LPS-induced ALI in rats were studied. ? Nilotinib showed potent anti-inflammatory activity as it attenuated PMN infiltration and hence ROS generation. ? In addition, nilotinib caused down-regulation of proinflammatory cytokine production.

387

PPAR? downregulates airway inflammation induced by lipopolysaccharide in the mouse  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Inflammation is a hallmark of acute lung injury and chronic airway diseases. In chronic airway diseases, it is associated with profound tissue remodeling. Peroxisome proliferator-activated receptor-? (PPAR? is a ligand-activated transcription factor, that belongs to the nuclear receptor family. Agonists for PPAR? have been recently shown to reduce lipopolysaccharide (LPS- and cytokine-induced secretion of matrix metalloproteinase-9 (MMP-9 in human monocytes and rat mesangial cells, suggesting that PPAR? may play a beneficial role in inflammation and tissue remodeling. Methods We have investigated the role of PPAR? in a mouse model of LPS-induced airway inflammation characterized by neutrophil and macrophage infiltration, by production of the chemoattractants, tumor necrosis factor-? (TNF-?, keratinocyte derived-chemokine (KC, macrophage inflammatory protein-2 (MIP-2 and monocyte chemoattractant protein-1 (MCP-1, and by increased MMP-2 and MMP-9 activity in bronchoalveolar lavage fluid (BALF. The role of PPAR? in this model was studied using both PPAR?-deficient mice and mice treated with the PPAR? activator, fenofibrate. Results Upon intranasal exposure to LPS, PPAR?-/- mice exhibited greater neutrophil and macrophage number in BALF, as well as increased levels of TNF-?, KC, MIP-2 and MCP-1, when compared to PPAR?+/+ mice. PPAR?-/- mice also displayed enhanced MMP-9 activity. Conversely, fenofibrate (0.15 to 15 mg/day dose-dependently reduced the increase in neutrophil and macrophage number induced by LPS in wild-type mice. In animals treated with 15 mg/day fenofibrate, this effect was associated with a reduction in TNF-?, KC, MIP-2 and MCP-1 levels, as well as in MMP-2 and MMP-9 activity. PPAR?-/- mice treated with 15 mg/day fenofibrate failed to exhibit decreased airway inflammatory cell infiltrate, demonstrating that PPAR? mediates the anti-inflammatory effect of fenofibrate. Conclusion Using both genetic and pharmacological approaches, our data clearly show that PPAR? downregulates cell infiltration, chemoattractant production and enhanced MMP activity triggered by LPS in mouse lung. This suggests that PPAR? activation may have a beneficial effect in acute or chronic inflammatory airway disorders involving neutrophils and macrophages.

Frossard Nelly

2005-08-01

388

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

Lifescience Database Archive (English)

Full Text Available 12106784 Structural and functional analyses of bacterial lipopolysaccharides. Caroff M, Karibian ... vaillon JM, Haeffner-Cavaillon N. Microbes Infect. 2002 ... Jul;4(9):915-26. (.png) (.svg) (.html) (.csml) Sho ... Haeffner-Cavaillon N. Publication Microbes Infect. 2002 ... Jul;4(9):915-26. Pathway - PNG File (.png) SVG Fil ...

389

Garlic (Allium sativum) Extracts Inhibits Lipopolysaccharide-Induced Toll-Like Receptor 4 Dimerization  

Science.gov (United States)

Garlic has been used as a folk medicine for a long history. Numerous studies demonstrated that garlic extracts and its sulfur-containing compounds inhibit nuclear factor-kappa B (NF-kB) activation induced by various receptor agonist including lipopolysaccharide (LPS). These effects suggest that garl...

390

ELEVATED MILK SOLUBLE CD14 IN BOVINE MAMMARY GLANDS CHALLENGED WITH ESCHERICHIA COLI LIPOPOLYSACCHARIDE  

Science.gov (United States)

The purpose of this study was to determine whether soluble CD14 in milk was affected by stage of lactation, milk somatic cell count (SCC), presence of bacteria or lipopolysaccharide (LPS)-induced inflammation. Milk samples from 100 lactating cows were assayed for sCD14 in milk to determine effects o...

391

DMPD: CD14 and other recognition molecules for lipopolysaccharide: a review. [Dynamic Macrophage Pathway CSML Database  

Lifescience Database Archive (English)

Full Text Available 7542643 CD14 ... and other recognition molecules for lipopolysaccharide: a review. Kielian TL, Blech ... ;29(3):187-205. (.png) (.svg) (.html) (.csml) Show CD14 ... and other recognition molecules for lipopolysaccha ... ride: a review. PubmedID 7542643 Title CD14 ... and other recognition molecules for lipopolysaccha ...

392

Alpha-lipoic acid protects mitochondrial enzymes and attenuates lipopolysaccharide-induced hypothermia in mice  

Science.gov (United States)

Abstract: Hypothermia is a key symptom of sepsis and the mechanism(s) leading to hypothermia during sepsis is largely unknown. To investigate a potential mechanism and find an effective treatment for hypothermia in sepsis, we induced hypothermia in mice by lipopolysaccharide (LP...

393

Differential Biofilm Formation and Motility Associated with Lipopolysaccharide/Exopolysaccharide-Coupled Biosynthetic Genes in Stenotrophomonas maltophilia  

OpenAIRE

Stenotrophomonas maltophilia WR-C is capable of forming biofilm on polystyrene and glass. The lipopolysaccharide/exopolysaccharide-coupled biosynthetic genes rmlA, rmlC, and xanB are necessary for biofilm formation and twitching motility. Mutants with mutations in rmlAC and xanB display contrasting biofilm phenotypes on polystyrene and glass and differ in swimming motility.

Huang, Tzu-pi; Somers, Eileen B.; Wong, Amy C. Lee

2006-01-01

394

Ferritin stimulation of a monokine inhibitor of lipopolysaccharide-augmented myelopoiesis is ferroxidase dependent.  

OpenAIRE

Ferritin inhibition of myelopoiesis has been associated with intrinsic ferroxidase activity of heavy-chain ferritin and with production of a monokine inhibitor of lipopolysaccharide (LPS)-augmented monocytopoiesis. We report here that intrinsic ferroxidase activity of heavy-chain ferritin is required for stimulated production of the monokine inhibitor of LPS-augmented monocytopoiesis.

Kreisberg, R.; Broxmeyer, H. E.; Moore, R. N.

1994-01-01

395

Interaction of peptide-bound beads with lipopolysaccharide and lipoproteins.  

Science.gov (United States)

We previously reported the generation of lipopolysaccharide (LPS)-binding peptides by phage display and chemical modification. Among them, a dodecapeptide designated Li5-025 (K'YSSSISSIRAC'; K' and C' denote d-lysine and d-cysteine, respectively) showed a high binding affinity for LPS and was resistant to protease digestion (Suzuki et al., 2010). In the current study, Li5-025-bound silica beads, hereafter referred to as P-beads, were generated and found to be devoid of LPS-neutralizing activity. Thus, LPS bound to the P-beads could be directly used in the Limulus amebocyte lysate (LAL) assay. P-beads bound LPS dissolved in solutions of ethanol, pH4, pH10, and 0.5M NaCl and LPS bound to the P-beads was quantitatively assayed. The sensitivity of this assay was observed to be approximately 0.1pg/mL LPS. P-beads bound LPS dissolved in antithrombin III (AT III) solution which is a strong inhibitor of activated factors C and B as well as the clotting enzyme in the LAL assay; the inhibitory effect of AT III was completely reversed upon washing the P-beads with 25% acetonitrile. This was employed as the first step for the detection of free LPS in plasma using the LAL assay. LPS added to human plasma at 0°C followed by application to the P-beads and subsequent washing with 25% acetonitrile resulted in low LPS activity as detected by the LAL assay. However, further washing of the P-beads with 0.1% Triton X100 in 25% acetonitrile resulted in high LPS activity. This is the first instance of quantitative detection of free LPS in plasma using the LAL assay, and the sensitivity of this method was observed to be 1pg/mL of LPS. The proteins eluted in the 0.1% Triton X-100 wash were analyzed using sodium dodecyl sulfate polyacrylamide gel electrophoresis. Two protein bands of 28kDa and 18kDa were predominantly observed. Mass spectrometry analysis revealed that the 28kDa and 18kDa bands corresponded to apolipoprotein A-I (apoA-I) and apolipoprotein A-II (apoA-II), respectively. ApoA-I and apoA-II are components of high density lipoprotein (HDL). Thus, it is likely that the P-beads-bound LPS was sequ