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Sample records for porphyromonas gingivalis lipopolysaccharide

  1. Effect of Nicotine and Porphyromonas gingivalis Lipopolysaccharide on Endothelial Cells In Vitro

    OpenAIRE

    An, Na; Andrukhov, Oleh; Tang, Yan; Falkensammer, Frank; Bantleon, Hans-peter; Ouyang, Xiangying; Rausch-fan, Xiaohui

    2014-01-01

    Smoking is considered a significant risk factor for both periodontal disease and cardiovascular disease (CVD). Endothelial cells play an important role in the progression of both diseases. In the present study, we investigated in vitro the impact of nicotine on functional properties of human umbilical vein endothelial cells (HUVECs) stimulated with lipopolysaccharide (LPS) of periodontal pathogen Porphyromonas gingivalis. HUVECs were stimulated with different concentrations of nicotine (10 µ...

  2. 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)

    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

    Full Text Available 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.

  3. 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)

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

    2008-12-01

    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.

  4. 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)

    DIEGO GUALTERO

    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.

  5. Porphyromonas gingivalis: a clonal pathogen?

    OpenAIRE

    Morten Enersen

    2011-01-01

    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...

  6. Porphyromonas gingivalis: a clonal pathogen?

    Directory of Open Access Journals (Sweden)

    Morten Enersen

    2011-11-01

    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.

  7. Inhibition of Porphyromonas gingivalis Biofilm by Oxantel?

    OpenAIRE

    Dashper, Stuart; Ang, Ching-Seng; Liu, Sze Wei; Paolini, Rita; Veith, Paul; Reynolds, Eric

    2009-01-01

    Porphyromonas gingivalis is a major pathogen of chronic periodontitis and exists in a biofilm on the surface of the tooth root. Oxantel, a cholinergic anthelmintic and fumarate reductase inhibitor, significantly inhibited biofilm formation by P. gingivalis and disrupted established biofilms at concentrations below its MIC against planktonic cells. Oxantel was more effective against P. gingivalis in biofilm than metronidazole, a commonly used antibiotic for periodontitis.

  8. Anti-Inflammatory Effect of Heme Oxygenase-1 Toward Porphyromonas gingivalis Lipopolysaccharide in Macrophages Exposed to Gomisins A, G, and J

    OpenAIRE

    Ryu, Eun Yeon; Park, Sun Young; Kim, Sun Gun; Park, Da Jung; Kang, Jum Soon; Kim, Young Hun; Seetharaman, Rajaseker; Choi, Young-Whan; Lee, Sang-Joon

    2011-01-01

    Periodontitis, a chronic inflammatory periodontal disease that develops from gingivitis, is caused by periodontal pathogenic bacteria such as Porphyromonas gingivalis. Recent studies have focused on the antioxidant, anti–human immunodeficiency virus, anticarcinogenic, and anti-inflammatory properties of gomisins. However, the anti-inflammatory activities of gomisin plants through heme oxygenase-1 (HO-1) signals remain poorly defined. We found that gomisins' anti-inflammatory activity occurs v...

  9. Porphyromonas gingivalis Fimbriae Bind to Cytokeratin of Epithelial Cells

    OpenAIRE

    Sojar, Hakimuddin T.; Sharma, Ashu; Genco, Robert J.

    2002-01-01

    The adherence of Porphyromonas gingivalis to host cells is likely a prerequisite step in the pathogenesis of P. gingivalis-induced periodontal disease. P. gingivalis binds to and invades epithelial cells, and fimbriae are shown to be involved in this process. Little is known regarding epithelial receptor(s) involved in binding of P. gingivalis fimbriae. Using an overlay assay with purified P. gingivalis fimbriae as a probe, two major epithelial cell proteins with masses of 50 and 40 kDa were ...

  10. Presence of Porphyromonas gingivalis in gingival squamous cell carcinoma

    OpenAIRE

    Katz, Joseph; Onate, Mairelys D; Kaleb M. Pauley; Bhattacharyya, Indraneel; Cha, Seunghee

    2011-01-01

    Periodontal disease has been recently linked to a variety of systemic conditions such as diabetes, cardiovascular disease, preterm delivery, and oral cancer. The most common bacteria associated with periodontal disease, Porphyromonas gingivalis (P. gingivalis) has not yet been studied in the malignant gingival tissues. The objective of this study was to investigate the presence of P. gingivalis in specimens from squamous cell carcinoma patients. We have performed immunohistochemical staining ...

  11. Arginine deiminase inhibits Porphyromonas gingivalis surface attachment.

    Science.gov (United States)

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

    2013-02-01

    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

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

    OpenAIRE

    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

    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...

  13. Multilocus sequence analysis of Porphyromonas gingivalis indicates frequent recombination.

    Science.gov (United States)

    Koehler, Andreas; Karch, Helge; Beikler, Thomas; Flemmig, Thomas F; Suerbaum, Sebastian; Schmidt, Herbert

    2003-09-01

    In this study, the genetic relationship of 19 Porphyromonas gingivalis isolates from patients with periodontitis was investigated by multilocus sequence analysis. Internal 400-600 bp DNA fragments of the 10 chromosomal genes ef-tu, ftsQ, hagB, gpdxJ, pepO, mcmA, dnaK, recA, pga and nah were amplified by PCR and sequenced. No two isolates were identical at all 10 loci. Phylogenetic analyses indicated a panmictic population structure of P. gingivalis. Split decomposition analysis, calculation of homoplasy ratios and analyses of clustered polymorphisms all indicate that recombination plays a major role in creating the genetic heterogeneity of P. gingivalis. A standardized index of association of 0.0898 indicates that the P. gingivalis genes analysed are close to linkage equilibrium. PMID:12949166

  14. Porphyromonas gingivalis and Treponema denticola Exhibit Metabolic Symbioses

    OpenAIRE

    Tan, Kheng H.; Seers, Christine A.; Dashper, Stuart G.; Mitchell, Helen L.; Pyke, James S.; Meuric, Vincent; Slakeski, Nada; Cleal, Steven M.; Chambers, Jenny L.; Mcconville, Malcolm J.; Reynolds, Eric C.

    2014-01-01

    Porphyromonas gingivalis and Treponema denticola are strongly associated with chronic periodontitis. These bacteria have been co-localized in subgingival plaque and demonstrated to exhibit symbiosis in growth in vitro and synergistic virulence upon co-infection in animal models of disease. Here we show that during continuous co-culture a P. gingivalis:T. denticola cell ratio of 6?1 was maintained with a respective increase of 54% and 30% in cell numbers when compared with mono-culture. Co-c...

  15. TRANSMISSION OF Porphyromonas gingivalis FROM CAREGIVERS TO CHILDREN.

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    Assya Krasteva

    2012-03-01

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

  16. Assessment of Internalization and Viability of Porphyromonas gingivalis in KB Epithelial Cells by Confocal Microscopy

    OpenAIRE

    Houalet-Jeanne, S.; Pellen-Mussi, P; Tricot-Doleux, S.; Apiou, J.; Bonnaure-Mallet, M.

    2001-01-01

    Porphyromonas gingivalis (P. gingivalis) is considered to be one of the main periodontal pathogens. The goal of this work was to confirm the ability of P. gingivalis to invade host cells. We detected P. gingivalis inside KB cells by confocal microscopy and analyzed the various aspects of the adherence and internalization process. Lysates of P. gingivalis-infected KB cells were also examined using anaerobic growth techniques. The results showed the viability and ability to replicate, inside th...

  17. Identification of essential genes of the periodontal pathogen Porphyromonas gingivalis

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    Klein Brian A

    2012-10-01

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

  18. Citrullination and proteolytic processing of chemokines by Porphyromonas gingivalis.

    Science.gov (United States)

    Moelants, Eva A V; Loozen, Gitte; Mortier, Anneleen; Martens, Erik; Opdenakker, Ghislain; Mizgalska, Danuta; Szmigielski, Borys; Potempa, Jan; Van Damme, Jo; Teughels, Wim; Proost, Paul

    2014-06-01

    The outgrowth of Porphyromonas gingivalis within the inflammatory subgingival plaque is associated with periodontitis characterized by periodontal tissue destruction, loss of alveolar bone, periodontal pocket formation, and eventually, tooth loss. Potential virulence factors of P. gingivalis are peptidylarginine deiminase (PPAD), an enzyme modifying free or peptide-bound arginine to citrulline, and the bacterial proteases referred to as gingipains (Rgp and Kgp). Chemokines attract leukocytes during inflammation. However, posttranslational modification (PTM) of chemokines by proteases or human peptidylarginine deiminases may alter their biological activities. Since chemokine processing may be important in microbial defense mechanisms, we investigated whether PTM of chemokines by P. gingivalis enzymes occurs. Upon incubation of interleukin-8 (IL-8; CXCL8) with PPAD, only minor enzymatic citrullination was detected. In contrast, Rgp rapidly cleaved CXCL8 in vitro. Subsequently, different P. gingivalis strains were incubated with the chemokine CXCL8 or CXCL10 and their PTMs were investigated. No significant CXCL8 citrullination was detected for the tested strains. Interestingly, although considerable differences in the efficiency of CXCL8 degradation were observed with full cultures of various strains, similar rates of chemokine proteolysis were exerted by cell-free culture supernatants. Sequencing of CXCL8 incubated with supernatant or bacteria showed that CXCL8 is processed into its more potent forms consisting of amino acids 6 to 77 and amino acids 9 to 77 (the 6-77 and 9-77 forms, respectively). In contrast, CXCL10 was entirely and rapidly degraded by P. gingivalis, with no transient chemokine forms being observed. In conclusion, this study demonstrates PTM of CXCL8 and CXCL10 by gingipains of P. gingivalis and that strain differences may particularly affect the activity of these bacterial membrane-associated proteases. PMID:24686061

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

    Directory of Open Access Journals (Sweden)

    Diego Fernando Gualtero Escobar

    2014-06-01

    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.

  20. Prevalence of Porphyromonas gingivalis fimA genotypes in Caucasians.

    Science.gov (United States)

    Beikler, Thomas; Peters, Ulrike; Prajaneh, Seangsome; Prior, Karola; Ehmke, Benjamin; Flemmig, Thomas Frank

    2003-10-01

    The aim of the present study was to determine the prevalence of Porphyromonas gingivalis fimA genotypes in Caucasian patients with periodontitis. A total of 102 patients harboring P. gingivalis subgingivally were enrolled into the study. Pooled subgingival plaque samples of the six most severely affected sites were taken and analysed by fimA-specific polymerase chain reaction (PCR) and restriction analysis. Moreover, 26 P. gingivalis isolates were analysed by sequence analysis of the fimA gene. Sequence analysis revealed five major fimA genotypes (fimA types I-V) and allowed further subtyping of fimA genotypes II and IV into two subgroups each. The overall prevalences of fimA genotypes as assessed by PCR and restriction analysis among the P. gingivalis-positive patients with periodontitis were: type I, 25.5%; type II, 38.2%; type III, 4.9%; type IV, 18.6%; type V, 3.9%; and non-typable, 6.9%. Two patients were colonized by both type II and type IV, or type III and type IV fimA genotypes, respectively. Patients harboring different fimA genotypes showed no significant difference in severity of periodontal disease, as assessed by pocket probing depth and bleeding on probing following adjustment for smoking habit and age. The results indicate that predominant fimA genotypes in Caucasian periodontitis patients are types I, II, and IV. However, there was no difference in the association of the various fimA genotypes with disease severity. PMID:12974681

  1. A streptococcal effector protein that inhibits Porphyromonas gingivalis biofilm development.

    Science.gov (United States)

    Christopher, Aaron B; Arndt, Annette; Cugini, Carla; Davey, Mary E

    2010-11-01

    Dental plaque formation is a developmental process involving cooperation and competition within a diverse microbial community, approximately 70?% of which is composed of an array of streptococci during the early stages of supragingival plaque formation. In this study, 79 cell-free culture supernatants from a variety of oral streptococci were screened to identify extracellular compounds that inhibit biofilm formation by the oral anaerobe Porphyromonas gingivalis strain 381. The majority of the streptococcal supernatants (61 isolates) resulted in lysis of P. gingivalis cells, and some (17 isolates) had no effect on cell viability, growth or biofilm formation. One strain, however, produced a supernatant that abolished biofilm formation without affecting growth rate. Analysis of this activity led to the discovery that a 48 kDa protein was responsible for the inhibition. Protein sequence identification and enzyme activity assays identified the effector protein as an arginine deiminase. To identify the mechanism(s) by which this protein inhibits biofilm formation, we began by examining the expression levels of genes encoding fimbrial subunits; surface structures known to be involved in biofilm development. Quantitative RT-PCR analysis revealed that exposure of P. gingivalis cells to this protein for 1 h resulted in the downregulation of genes encoding proteins that are the major subunits of two distinct types of thin, single-stranded fimbriae (fimA and mfa1). Furthermore, this downregulation occurred in the absence of arginine deiminase enzymic activity. Hence, our data indicate that P. gingivalis can sense this extracellular protein, produced by an oral streptococcus (Streptococcus intermedius), and respond by downregulating expression of cell-surface appendages required for attachment and biofilm development. PMID:20705665

  2. Natural Competence Is a Major Mechanism for Horizontal DNA Transfer in the Oral Pathogen Porphyromonas gingivalis

    OpenAIRE

    Tribble, Gena D.; Rigney, Todd W.; Dao, Doan-Hieu V.; Wong, Cindy T.; Kerr, Jennifer E; Taylor, Brendan E.; Pacha, Sara; Kaplan, Heidi B.

    2012-01-01

    Porphyromonas gingivalis is a Gram-negative anaerobe that resides exclusively in the human oral cavity. Long-term colonization by P. gingivalis requires the bacteria to evade host immune responses while adapting to the changing host physiology and alterations in the composition of the oral microflora. The genetic diversity of P. gingivalis appears to reflect the variability of its habitat; however, little is known about the molecular mechanisms generating this diversity. Previously, our res...

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

    Scientific Electronic Library Online (English)

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

    2014-08-01

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

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

    OpenAIRE

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

    2007-01-01

    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...

  5. Periodontal disease, Porphyromonas gingivalis serum antibody levels and orodigestive cancer mortality

    OpenAIRE

    Ahn, Jiyoung; Segers, Stephanie; Hayes, Richard B

    2012-01-01

    Periodontitis, the progressive loss of the alveolar bone around the teeth and the major cause of tooth loss in adults, is due to oral microorganisms, including Porphyromonas gingivalis. Periodontitis is associated with a local overly aggressive immune response and a spectrum of systemic effects, but the role of this condition in orodigestive cancers is unclear. We prospectively examined clinically ascertained periodontitis (N = 12?605) and serum IgG immune response to P.gingivalis (N = 7852) ...

  6. Isolation and characterization of a cloned Porphyromonas gingivalis hemagglutinin from an avirulent strain of Salmonella typhimurium.

    OpenAIRE

    Dusek, D M; Progulske-Fox, A; Whitlock, J; Brown, T. A.

    1993-01-01

    Identification of surface macromolecules of Porphyromonas gingivalis that act as virulence factors in periodontal disease has important implications for studying host-parasite interactions as well as for potential vaccine development. The objective of this study was to determine whether a cloned, P. gingivalis hemagglutinin gene could be expressed in an intact form in an avirulent Salmonella typhimurium vaccine construct and to characterize the recombinant protein. The recombinant protein was...

  7. Cleavage of Human Transferrin by Porphyromonas gingivalis Gingipains Promotes Growth and Formation of Hydroxyl Radicals

    OpenAIRE

    Goulet, Ve?ronique; Britigan, Bradley; Nakayama, Koji; Grenier, Daniel

    2004-01-01

    Porphyromonas gingivalis, a gram-negative anaerobic bacterium associated with active lesions of chronic periodontitis, produces several proteinases which are presumably involved in host colonization, perturbation of the immune system, and tissue destruction. The aims of this study were to investigate the degradation of human transferrin by gingipain cysteine proteinases of P. gingivalis and to demonstrate the production of toxic hydroxyl radicals (HO·) catalyzed by the iron-containing transf...

  8. Metabolic Pathways for Cytotoxic End Product Formation from Glutamate- and Aspartate-Containing Peptides by Porphyromonas gingivalis

    OpenAIRE

    Takahashi, Nobuhiro; Sato, Takuichi; Yamada, Tadashi

    2000-01-01

    Metabolic pathways involved in the formation of cytotoxic end products by Porphyromonas gingivalis were studied. The washed cells of P. gingivalis ATCC 33277 utilized peptides but not single amino acids. Since glutamate and aspartate moieties in the peptides were consumed most intensively, a dipeptide of glutamate or aspartate was then tested as a metabolic substrate of P. gingivalis. P. gingivalis cells metabolized glutamylglutamate to butyrate, propionate, acetate, and ammonia, and they met...

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

    OpenAIRE

    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

    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...

  10. Porphyromonas gingivalis: keeping the pathos out of the biont.

    Science.gov (United States)

    Cugini, Carla; Klepac-Ceraj, Vanja; Rackaityte, Elze; Riggs, James E; Davey, Mary E

    2013-01-01

    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'. PMID:23565326

  11. NOD2 contributes to Porphyromonas gingivalis-induced bone resorption.

    Science.gov (United States)

    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

    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

  12. Porphyromonas gingivalis: keeping the pathos out of the biont

    Directory of Open Access Journals (Sweden)

    Carla Cugini

    2013-04-01

    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’.

  13. Periodontitis and Porphyromonas gingivalis in Preclinical Stage of Arthritis Patients

    Science.gov (United States)

    Hashimoto, Motomu; Yamazaki, Toru; Hamaguchi, Masahide; Morimoto, Takeshi; Yamori, Masashi; Asai, Keita; Isobe, Yu; Furu, Moritoshi; Ito, Hiromu; Fujii, Takao; Terao, Chikashi; Mori, Masato; Matsuo, Takashi; Yoshitomi, Hiroyuki; Yamamoto, Keiichi; Yamamoto, Wataru; Bessho, Kazuhisa; Mimori, Tsuneyo

    2015-01-01

    Purpose To determine whether the presence of periodontitis (PD) and Porphyromonas gingivalis (Pg) in the subgingival biofilm associates with the development of rheumatoid arthritis (RA) in treatment naïve preclinical stage of arthritis patients. Methods We conducted a prospective cohort study of 72 consecutive patients with arthralgia who had never been treated with any anti-rheumatic drugs or glucocorticoids. Periodontal status at baseline was assessed by dentists. PD was defined stringently by the maximal probing depth?4 mm, or by the classification by the 5th European Workshop in Periodontology (EWP) in 2005 using attachment loss. Up to eight plaque samples were obtained from each patient and the presence of Pg was determined by Taqman PCR. The patients were followed up for 2 years and introduction rate of methotrexate (MTX) treatment on the diagnosis of RA was compared in patients with or without PD or Pg. Results Patients with PD (probing depth?4mm) had higher arthritis activity (p = 0.02) and higher risk for future introduction of MTX treatment on the diagnosis of RA during the follow up than patients without PD (Hazard ratio 2.68, p = 0.03). Arthritis activity and risk for MTX introduction increased with the severity of PD assessed by EWP, although not statistically significant. On the other hand, presence of Pg was not associated with arthritis activity (p = 0.72) or the risk for MTX introduction (p = 0.45). Conclusion In treatment naïve arthralgia patients, PD, but not the presence of Pg, associates with arthritis activity and future requirement of MTX treatment on the diagnosis of RA. PMID:25849461

  14. Effect of simulated high-altitude hypoxia on Porphyromonas gingivalis

    Directory of Open Access Journals (Sweden)

    Jing-jing HUANG

    2012-04-01

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

  15. Activation of the NLRP3 inflammasome in Porphyromonas gingivalis-accelerated atherosclerosis.

    Science.gov (United States)

    Yamaguchi, Yohei; Kurita-Ochiai, Tomoko; Kobayashi, Ryoki; Suzuki, Toshihiko; Ando, Tomohiro

    2015-06-01

    Porphyromonas gingivalis has been shown to accelerate atherosclerotic lesion development in hyperlipidemic animals. Atherosclerosis is a disease characterized by inflammation of the arterial wall. Recent studies have suggested that the NLRP3 inflammasome plays an important role in the development of vascular inflammation and atherosclerosis. Herein, we investigated a possible association between the inflammasome in atherosclerosis and periodontal disease induced by P. gingivalis infection using apolipoprotein E-deficient, spontaneously hyperlipidemic (Apoe(shl)) mice. Oral infection with wild-type (WT) P. gingivalis significantly increased the area of aortic sinus covered with atherosclerotic plaque and alveolar bone loss, compared with KDP136 (gingipain-null mutant) or KDP150 (FimA-deficient mutant) challenge. WT challenge also increased IL-1?, IL-18 and TNF-? production in peritoneal macrophages, and gingival or aortic gene expression of Nod-like receptor family, pyrin domain containing 3 (NLRP3), pro-IL-1?, pro-IL-18 and pro-caspase-1. Porphyromonas gingivalis genomic DNA was detected more in the aorta, gingival tissue, liver and spleen of WT-challenged mice than those in KDP136- or KDP150-challenged mice. We conclude that WT P. gingivalis activates innate immune cells through the NLRP3 inflammasome compared with KDP136 or KDP150. The NLRP3 inflammasome may play a critical role in periodontal disease and atherosclerosis induced by P. gingivalis challenge through sustained inflammation. PMID:25663345

  16. Association of the invasion ability of Porphyromonas gingivalis with the severity of periodontitis.

    Science.gov (United States)

    Baek, Keum Jin; Ji, Suk; Kim, Yong Chul; Choi, Youngnim

    2015-01-01

    Porphyromonas gingivalis is one of the well-characterized periodontal pathogens involved in periodontitis. The invasive and proteolytic activities of P. gingivalis clinical isolates have been shown to be associated with heterogenic virulence, as determined in a mouse abscess model. The aims of the present study were to identify a P. gingivalis strain with a low virulence among clinical isolates, based on its invasive ability and cytokine proteolytic activities, and to explore the preferential degradation of a certain cytokine by P. gingivalis. P. gingivalis ATCC 33277, W50, and 10 clinical isolates were used. After incubating bacteria with IL-4, IL-6, IL-10, IL-17A, TNF?, IFN?, and IL-1?, the amounts of remaining cytokines were determined by ELISA. Invasion ability was measured by a flow cytometric invasion assay. There was inter-strain variability both in the cytokine proteolytic activities and invasion ability. In addition, differential degradation of cytokines by P. gingivalis was observed: while IFN? and IL-17A were almost completely degraded, inflammatory cytokines TNF? and IL-1? were less susceptible to degradation. Interestingly, the invasion index, but not cytokine proteolytic activities, of P. gingivalis had strong positive correlations with clinical parameters of subjects who harbored the isolates. Therefore, the invasive ability of P. gingivalis is an important virulence factor, and the bacterial invasion step may be a good target for new therapeutics of periodontitis. PMID:25616643

  17. Heightened immune response to autocitrullinated Porphyromonas gingivalis peptidylarginine deiminase: a potential mechanism for breaching immunologic tolerance in rheumatoid arthritis

    OpenAIRE

    Quirke, AM; Lugli, EB; Wegner, N; Hamilton, BC; Charles, P.; Chowdhury, M; Ytterberg, AJ; Zubarev, RA; Potempa, J.; Culshaw, S; Guo, Y.; Fisher, BA; Thiele, G.; Mikuls, TR; Venables, PJ

    2013-01-01

    Background: Rheumatoid arthritis (RA) is characterised by autoimmunity to citrullinated proteins, and there is increasing epidemiologic evidence linking Porphyromonas gingivalis to RA. P gingivalis is apparently unique among periodontal pathogens in possessing a citrullinating enzyme, peptidylarginine deiminase (PPAD) with the potential to generate antigens driving the autoimmune response. Objectives: To examine the immune response to PPAD in patients with RA, individuals with periodontit...

  18. The K1 Serotype Capsular Polysaccharide of Porphyromonas gingivalis Elicits Chemokine Production from Murine Macrophages That Facilitates Cell Migration ?

    OpenAIRE

    d'Empaire, Gabriela; Baer, Michael T.; Gibson, Frank C

    2006-01-01

    Porphyromonas gingivalis is the principal organism associated with aggressive forms of generalized periodontal disease. Previous reports have suggested that encapsulated P. gingivalis strains are more virulent than unencapsulated strains; however, the contribution of capsular polysaccharide (CPS) to the virulence of this organism is poorly understood. Since periodontal disease presents with a complex inflammatory cell lesion comprised of neutrophils and monocytes, we cultured murine peritonea...

  19. Roles of porphyrins and host iron transport proteins in regulation of growth of Porphyromonas gingivalis W50.

    OpenAIRE

    Bramanti, T E; Holt, S. C.

    1991-01-01

    Porphyromonas gingivalis (Bacteroides gingivalis) requires iron in the form of hemin for growth and virulence in vitro, but the contributions of the porphyrin ring structure, porphyrin-associated iron, host hemin-sequestering molecules, and host iron-withholding proteins to its survival are unknown. Therefore, the effects of various porphyrins, host iron transport proteins, and inorganic iron sources on the growth of P. gingivalis W50 were examined to delineate the various types of iron molec...

  20. Clonal diversity of the taxon Porphyromonas gingivalis assessed by random amplified polymorphic DNA fingerprinting.

    OpenAIRE

    Me?nard, C.; Mouton, C.

    1995-01-01

    A total of 97 strains of the periopathogen Porphyromonas gingivalis were collected. This collection included laboratory strains and clinical isolates of human origin with diverse clinical and geographical origins. Biological diversity was further increased by including 32 strains isolated from the oral cavities of nine different animal species. Genomic fingerprints of the 129 strains were generated as random amplified polymorphic DNAs (RAPDs) by the technique of PCR amplification with a singl...

  1. Transglutaminase 2 is essential for adherence of Porphyromonas gingivalis to host cells

    OpenAIRE

    Boisvert, Heike; Lorand, Laszlo; Duncan, Margaret J.

    2014-01-01

    Transglutaminase 2 (TG2) is widely distributed inside and outside cells, and is one of a family of nine proteins in the human genome that likely evolved from the papain group of cysteine proteases. Apart from its transamidating activity, TG2 is known to form tight association with fibronectin (FN), a universal component of ECM. New findings presented in our paper demonstrate that the TG2–FN complex mediates the attachment of Porphyromonas gingivalis, an oral bacterium and causative agent of...

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

    OpenAIRE

    Enersen, Morten

    2011-01-01

    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...

  3. Mechanism of methaemoglobin breakdown by the lysine-specific gingipain of the periodontal pathogen Porphyromonas gingivalis

    OpenAIRE

    Smalley, John W; Birss, Andrew J; Szmigielski, Borys; Potempa, Jan

    2008-01-01

    The R- and K-gingipain proteases of Porphyromonas gingivalis are involved in proteolysis of haemoglobin from which the defensive dimeric haem pigment is formed. Whilst oxyhaemoglobin is refractory towards K-gingipain, methaemoglobin is rapidly degraded. Ligation of methaemoglobin with N3?, which effectively blocks haem dissociation from the protein, prevented haemoglobin breakdown. Haem-free globin was rapidly degraded by K-gingipain. These data emphasise the need for haemoglobin oxidation wh...

  4. Inactivation of Epidermal Growth Factor by Porphyromonas gingivalis as a Potential Mechanism for Periodontal Tissue Damage

    OpenAIRE

    Pyrc, Krzysztof; Milewska, Aleksandra; Kantyka, Tomasz; Sroka, Aneta; Maresz, Katarzyna; Kozie?, Joanna; Nguyen, Ky-Anh; Jan J. Enghild; Knudsen, Anders Dahl; Potempa, Jan

    2013-01-01

    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 peptidylarginine deiminase (PPAD), an enzyme that 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 an...

  5. Purification, Characterization, and Sequence Analysis of a Potential Virulence Factor from Porphyromonas gingivalis, Peptidylarginine Deiminase

    OpenAIRE

    McGraw, Walker T.; Potempa, Jan; Farley, David; Travis, James

    1999-01-01

    The initiation and progression of adult-onset periodontitis has been associated with infection of the gingival sulcus by Porphyromonas gingivalis. This organism utilizes a multitude of virulence factors to evade host defenses as it establishes itself as one of the predominant pathogens in periodontal pockets. A feature common to many other oral pathogens is the production of ammonia due to its protective effect during acidic cleansing cycles in the mouth. Additionally, ammonia production by P...

  6. HU Protein Affects Transcription of Surface Polysaccharide Synthesis Genes in Porphyromonas gingivalis ? †

    OpenAIRE

    Alberti-segui, Christine; Arndt, Annette; Cugini, Carla; Priyadarshini, Richa; Davey, Mary E.

    2010-01-01

    K-antigen capsule synthesis is an important virulence determinant of the oral anaerobe Porphyromonas gingivalis. We previously reported that the locus required for synthesis of this surface polysaccharide in strain W83 (TIGR identification PG0106 to PG0120) is transcribed as a large (?16.7-kb) polycistronic message. Through sequence analysis, we have now identified a 77-bp inverted repeat located upstream (206 bp) of the start codon of PG0106 that is capable of forming a large hairpin struc...

  7. Asociación de la severidad de la periodontitis con niveles de cotinina y Porphyromonas gingivalis

    Directory of Open Access Journals (Sweden)

    Carlos Mart\\u00EDn Ardila Medina

    2014-01-01

    Full Text Available Fundamento: la cotinina aumenta los efectos de las toxinas producidas por los periodontopatógenos y se ha observado que el hábito de fumar altera la respuesta humoral e incrementa la infectividad de la Porphyromonas gingivalis. Objetivo: investigar la asociación entre los niveles de cotinina, la severidad y la extensión de la periodontitis, entre los niveles de cotinina y presencia de P. gingivalis. Método: en el presente estudio de corte transversal, el universo estuvo constituido por 108 sujetos. Los parámetros periodontales se midieron en seis sitios por diente en todos los dientes, se excluyó el tercer molar. Se tomaron muestras de P. gingivalis en las bolsas periodontales. Resultados: al comparar fumadores y no fumadores se observaron diferencias estadísticamente significativas en la profundidad de sondaje y en el nivel de inserción clínica, con peores condiciones periodontales en los fumadores (p < 0.001. Se encontró P. gingivalis en 64 sujetos (59, 3 % y niveles de cotinina ? 10 ng/ml en 25 pacientes (23, 1 %. Se observó una asociación estadísticamente significativa entre periodontitis avanzada y niveles de cotinina ? 10 ng/ml (p < 0.001, y entre niveles de cotinina ? 10 ng/ml y presencia de P. gingivalis (p < 0.05. Conclusiones: los niveles de cotinina en suero ? 10 ng/ml se asociaron con bolsas periodontales más profundas y mayor pérdida de inserción; igualmente se encontró asociación entre cotinina y P. gingivalis,con peores condiciones clínicas periodontales en los sujetos fumadores.

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

    Science.gov (United States)

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

    2014-01-01

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

  9. Porphyromonas gingivalis and Treponema denticola Mixed Microbial Infection in a Rat Model of Periodontal Disease

    Directory of Open Access Journals (Sweden)

    Raj K. Verma

    2010-01-01

    Full Text Available Porphyromonas gingivalis and Treponema denticola are periodontal pathogens that express virulence factors associated with the pathogenesis of periodontitis. In this paper we tested the hypothesis that P. gingivalis and T. denticola are synergistic in terms of virulence; using a model of mixed microbial infection in rats. Groups of rats were orally infected with either P. gingivalis or T. denticola or mixed microbial infections for 7 and 12 weeks. P. gingivalis genomic DNA was detected more frequently by PCR than T. denticola. Both bacteria induced significantly high IgG, IgG2b, IgG1, IgG2a antibody levels indicating a stimulation of Th1 and Th2 immune response. Radiographic and morphometric measurements demonstrated that rats infected with the mixed infection exhibited significantly more alveolar bone loss than shaminfected control rats. Histology revealed apical migration of junctional epithelium, rete ridge elongation, and crestal alveolar bone resorption; resembling periodontal disease lesion. These results showed that P. gingivalis and T. denticola exhibit no synergistic virulence in a rat model of periodontal disease.

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

    DEFF Research Database (Denmark)

    BelstrØm, Daniel; Holmstrup, Palle

    2011-01-01

    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.

  11. 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)

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

    2012-12-01

    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.

  12. 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)

    Sandra, Moreno; Adolfo, Contreras.

    2013-01-01

    Full Text Available 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.

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

    Scientific Electronic Library Online (English)

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

    2010-04-01

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

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

    Scientific Electronic Library Online (English)

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

    2007-06-01

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

  15. Cloning of the gene encoding a glycylprolyl aminopeptidase from Porphyromonas gingivalis.

    Science.gov (United States)

    Nakamura, S; Takeuchi, A; Masamoto, Y; Abiko, Y; Hayakawa, M; Takiguchi, H

    1992-10-01

    A genomic library of Porphyromonas gingivalis 381 was constructed in the cosmid vector pHC79. A clone, pSN1, was identified by the expression of glycylprolyl-naphthylamide hydrolysing activity. The DNA insert contained within the cosmid pSN1 was subcloned into the plasmid vector pBR328 to create the recombinant plasmid pSN11 containing a 2.9 kb EcoRV insert. An Escherichia coli transformant containing pSN11 produced a protein having a molecular weight of 75 kDa. Southern-blot hybridization revealed that the 2.9 kb EcoRV DNA hybridized with an identical sized Eco RV DNA fragment in the chromosomal DNA of P. gingivalis 381. PMID:1332661

  16. The nucleoid-associated protein HU? affects global gene expression in Porphyromonas gingivalis

    OpenAIRE

    Priyadarshini, Richa; Cugini, Carla; Arndt, Annette; Chen, Tsute; Tjokro, Natalia O.; Goodman, Steven D.; Davey, Mary E.

    2013-01-01

    HU is a non-sequence-specific DNA-binding protein and one of the most abundant nucleoid-associated proteins in the bacterial cell. Like Escherichia coli, the genome of Porphyromonas gingivalis is predicted to encode both the HU? (PG1258) and the HU? (PG0121) subunit. We have previously reported that PG0121 encodes a non-specific DNA-binding protein and that PG0121 is co-transcribed with the K-antigen capsule synthesis operon. We also reported that deletion of PG0121 resulted in downregulati...

  17. Isolation and characterization of a cloned Porphyromonas gingivalis hemagglutinin from an avirulent strain of Salmonella typhimurium.

    Science.gov (United States)

    Dusek, D M; Progulske-Fox, A; Whitlock, J; Brown, T A

    1993-03-01

    Identification of surface macromolecules of Porphyromonas gingivalis that act as virulence factors in periodontal disease has important implications for studying host-parasite interactions as well as for potential vaccine development. The objective of this study was to determine whether a cloned, P. gingivalis hemagglutinin gene could be expressed in an intact form in an avirulent Salmonella typhimurium vaccine construct and to characterize the recombinant protein. The recombinant protein was purified from the vaccine strain, characterized, and tested for biological activity as a competitive inhibitor of hemagglutination. Cells of S. typhimurium SL3261/pST7 grown in Luria broth were broken by sonic disruption and fractionated. The purified recombinant protein was found to inhibit hemagglutination of erythrocytes by whole P. gingivalis cells. The same purified protein was analyzed for its N-terminal amino acid sequence and amino acid composition and found to match that predicted from the nucleotide sequence of the cloned gene. These results indicate that a surface macromolecule of P. gingivalis can be expressed in an intact and biologically active form in a Salmonella carrier strain. PMID:8381773

  18. Structures of the Porphyromonas gingivalis OxyR regulatory domain explain differences in expression of the OxyR regulon in Escherichia coli and P. gingivalis

    Energy Technology Data Exchange (ETDEWEB)

    Svintradze, David V. [Virginia Commonwealth University, Richmond, VA 23298-0566 (United States); Virginia Commonwealth University, Richmond, VA 23219-1540 (United States); Peterson, Darrell L. [Virginia Commonwealth University, Richmond, VA 23219-1540 (United States); Virginia Commonwealth University, Richmond, VA 23298-0614 (United States); Collazo-Santiago, Evys A.; Lewis, Janina P. [Virginia Commonwealth University, Richmond, VA 23298-0566 (United States); Wright, H. Tonie, E-mail: xrdproc@vcu.edu [Virginia Commonwealth University, Richmond, VA 23219-1540 (United States); Virginia Commonwealth University, Richmond, VA 23298-0614 (United States); Virginia Commonwealth University, Richmond, VA 23298-0566 (United States)

    2013-10-01

    Differences in OxyR regulated expression of oxidative stress genes between Escherichia coli and Porphyromonas gingivalis are explained by very minor differences in structure and amino-acid sequence of the respective oxidized and reduced OxyR regulatory domains. These differences affect OxyR quaternary structures and are predicted from model building of full length OxyR–DNA complexes to confer distinct modes of DNA binding on this transcriptional regulator. OxyR transcriptionally regulates Escherichia coli oxidative stress response genes through a reversibly reducible cysteine disulfide biosensor of cellular redox status. Structural changes induced by redox changes in these cysteines are conformationally transmitted to the dimer subunit interfaces, which alters dimer and tetramer interactions with DNA. In contrast to E. coli OxyR regulatory-domain structures, crystal structures of Porphyromonas gingivalis OxyR regulatory domains show minimal differences in dimer configuration on changes in cysteine disulfide redox status. This locked configuration of the P. gingivalis OxyR regulatory-domain dimer closely resembles the oxidized (activating) form of the E. coli OxyR regulatory-domain dimer. It correlates with the observed constitutive activation of some oxidative stress genes in P. gingivalis and is attributable to a single amino-acid insertion in P. gingivalis OxyR relative to E. coli OxyR. Modelling of full-length P. gingivalis, E. coli and Neisseria meningitidis OxyR–DNA complexes predicts different modes of DNA binding for the reduced and oxidized forms of each.

  19. Peptidyl arginine deiminase from Porphyromonas gingivalis abolishes C5a activity

    DEFF Research Database (Denmark)

    Bielecka, Ewa; Scavenius, Carsten

    2014-01-01

    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.

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

    Science.gov (United States)

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

    2013-05-01

    To evaluate the possibility of an alternative to the traditional orthodontic stainless steel implants, the antibacterial activity against Porphyromonas gingivalis (P. gingivalis) and the related cytotoxicity of a type 304 Cu bearing antibacterial stainless steel were studied. The results indicated that the antibacterial stainless steel showed excellent antibacterial property against P. gingivalis, compared with the control steel (a purchased medical grade 304 stainless steel). Compared to the control steel, there were fewer bacteria on the surface of the antibacterial stainless steel, with significant difference in morphology. The cytotoxicities of the antibacterial stainless steel to both MG-63 and KB cells were all grade 1, the same as those of the control steel. There were no significant differences in the apoptosis rates on MG-63 and KB cells between the antibacterial stainless steel and the control steel. This study demonstrates that the antibacterial stainless steel is possible to reduce the incidence of implant-related infections and can be a more suitable material for the micro-implant than the conventional stainless steel in orthodontic treatment. PMID:23352947

  1. Bactericidal effect of a 405-nm diode laser on Porphyromonas gingivalis

    International Nuclear Information System (INIS)

    The study was conducted to determine the effect of 405-nm diode laser irradiation on periodontopathic bacteria such as Porphyromonas gingivalis in vitro. A diluted suspension of P. gingivalis was irradiated directly with a 405-nm diode laser under conditions of 100 mW-10 sec, 100 mW-20 sec, 200 mW-5 sec, 200 mW-10 sec, 200 mW-20 sec, 400 mW-5 sec, 400 mW-10 sec, and 400 mW-20 sec. The energy density ranged from 2.0 to 16.0 J/cm2. The irradiated bacterial suspension was spread on a blood agar plate and growth of the colonies was examined after an anaerobic culture for 7 days. Bacterial growth was inhibited under all irradiation conditions, but the bactericidal effect of the 405-nm diode laser depended on the energy density. More than 97% of bacterial growth was inhibited with irradiation at an energy density > 4.0 J/cm2. The mechanism of the bactericidal effect is photochemical, rather than photothermal. These findings suggest that a 405-nm diode laser has a high bactericidal effect on P. gingivalis

  2. Sequence analysis of kgp in Porphyromonas gingivalis isolates from periodontitis patients.

    Science.gov (United States)

    Beikler, T; Peters, U; Ehmke, B; Flemmig, T F

    2003-12-01

    The aim of the present study was to determine sequence variation in the Lys-x-specific protease (Kgp) encoding gene kgp of Porphyromonas gingivalis and to analyze its association with periodontal disease severity. Pooled subgingival plaque samples were obtained from the six most severely affected sites of 102 patients with periodontitis. Sequence analysis of the kgp gene in 23 clinical P. gingivalis isolates resulted in the identification of two distinct kgp types (kgp-I and kgp-II) according to sequence differences in the region encoding the catalytic domain. Restriction analysis revealed that 59 of the 102 patients were colonized by kgp-I and 43 by kgp-II. Patients harboring kgp-I or kgp-II showed no significant difference in the severity of periodontal disease as assessed by pocket probing depth and bleeding on probing following adjustment for smoking habit and age. Moreover, no differences in proteolytic activity of Kgp-I and Kgp-II were detected. The results indicated that two kgp types are maintained in natural populations of P. gingivalis. PMID:14622346

  3. Blocking Pro-Inflammatory Cytokine Release Modulates Peripheral Blood Mononuclear Cell Response to Porphyromonas Gingivalis

    Science.gov (United States)

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

    2013-01-01

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

  4. 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)

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

    2010-12-01

    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.

  5. Isolation and characterization of transposon-induced mutants of Porphyromonas gingivalis deficient in fimbriation.

    Science.gov (United States)

    Watanabe-Kato, T; Hayashi, J I; Terazawa, Y; Hoover, C I; Nakayama, K; Hibi, E; Kawakami, N; Ikeda, T; Nakamura, H; Noguchi, T; Yoshimura, F

    1998-01-01

    Fimbriae are considered to be an important virulence factor of Porphyromonas gingivalis. In order to identify genes essential for fimbriation, other than fimA which encodes the major subunit protein of fimbriae, transposon mutagenesis and immunological screening techniques were used to isolate fimbria-deficient mutants. R751::*Omega4, a suicide vector that carries Tn4351, was transferred from Escherichia coli to P. gingivalis by conjugation. Twenty-two independent fimbria-deficient mutants were identified among the resulting transformants. Southern hybridization analysis with pBlue 4351, a transposon-specific probe, and R751 indicated that 45% of the mutants resulted from single transposon insertions and that the remaining 55% of the mutants resulted from cointegration of R751 sequences. Southern hybridization analysis with pUCBg12.1, a probe for the fimA region, indicated that nine of the mutants contained insertions within the 2.5 kb SacI DNA fragment of P. gingivalis that contains fimA, ORF1 (which encodes a 15 kDa protein), and the C-terminal portion of ORF5 (which encodes a 63 kDa protein). Polymerase chain reaction (PCR) analysis and further Southern hybridization analysis indicated that the insertion site(s) for all nine of these mutants was within the fimA gene. Southern hybridization analysis also indicated that the remaining thirteen mutants contained insertions somewhere outside the 10 kb fimA region. Analysis by pulsed field gel electrophoresis (PFGE) revealed that insertions for most of the thirteen mutants mapped to a 300 kb NotI fragment and are located at least approximately 200 kb away from fimA. These results identify genetic loci other than fimA, that are required for fimbriation of P. gingivalis. Future cloning and characterization of these genetic loci should be straightforward since they are now marked by antibiotic resistance genes carried by the transposon. PMID:9466944

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

    Directory of Open Access Journals (Sweden)

    Mônica Franca

    2007-06-01

    Full Text Available Introduction: Periodontitis is a chronic disease that results from an interaction of a mixed bacterial challenge and the host response. Objective: The purposes of this study were to evaluate the IgG serum levels to Porphyromonas gingivalis antigens by ELISA in individuals with different periodontal conditions correlated with clinical parameters, and to analyze the immunoreactivity profiles by Western blotting. Methods: Serum IgG levels against the cell sonicate antigen from P. gingivalis ATCC 33277 of 28 patients with chronic periodontitis (CP, 10 patients with gingivitis (G and 21 periodontally healthy individuals (H were measured by ELISA and Western immunoblotting. Results: In the CP group, sera reactivity by ELISA was significantly higher than in the G and H groups (Kruskal-Wallis p<0.001; Dunnet t3 p= 0.001 and Dunnet t3 p= 0.0001. There was no statistically significant difference between G and HP reactivity (Dunnett t3 p=0.617. Among individuals with chronic periodontitis, the IgG-anti-P. gingivalis serum levels were positively correlated with percentage of clinical attachment level =5mm (r s = + 0.375, p<0.05 and a negative correlation was found between IgG-anti-P. gingivalis levels and percentage of probing pocket depth 0-3mm (r s = - 0. 411, p< 0.05. The analysis of sera immunoreactivity profiles to sonicate antigen by Western blotting showed differences between the sera of CP, G and H group individuals. The serum from CP frequently reacted with high molecular weight (103 kDa, 86 kDa, 72 kDa, 60 kDa, 58 kDa, 52 kDa protein fractions. Conclusions: Serum levels of IgG anti-P. gingivalis distinguished individuals with chronic periodontitis, gingivitis and healthy periodontium. There was a correlation between clinical parameters and serum IgG levels against P. gingivalis. There was a difference in the recognition profile of protein fractions among the studied groups and some bands were more specific

  7. Heme environment in HmuY, the heme-binding protein of Porphyromonas gingivalis.

    Science.gov (United States)

    Wójtowicz, Halina; Wojaczy?ski, Jacek; Olczak, Mariusz; Króliczewski, Jaros?aw; Latos-Grazy?ski, Lechos?aw; Olczak, Teresa

    2009-05-29

    Porphyromonas gingivalis, a Gram-negative anaerobic bacterium implicated in the development and progression of chronic periodontitis, acquires heme for growth by a novel mechanism composed of HmuY and HmuR proteins. The aim of this study was to characterize the nature of heme binding to HmuY. The protein was expressed, purified and detailed investigations using UV-vis absorption, CD, MCD, and (1)H NMR spectroscopy were carried out. Ferric heme bound to HmuY may be reduced by sodium dithionite and re-oxidized by potassium ferricyanide. Heme complexed to HmuY, with a midpoint potential of 136mV, is in a low-spin Fe(III) hexa-coordinate environment. Analysis of heme binding to several single and double HmuY mutants with the methionine, histidine, cysteine, or tyrosine residues replaced by an alanine residue identified histidines 134 and 166 as potential heme ligands. PMID:19345198

  8. Heme environment in HmuY, the heme-binding protein of Porphyromonas gingivalis

    Energy Technology Data Exchange (ETDEWEB)

    Wojtowicz, Halina [Laboratory of Biochemistry, Faculty of Biotechnology, University of Wroclaw, Tamka 2, 50-137 Wroclaw (Poland); Wojaczynski, Jacek [Department of Chemistry, University of Wroclaw, 50-383 Wroclaw (Poland); Olczak, Mariusz [Laboratory of Biochemistry, Faculty of Biotechnology, University of Wroclaw, Tamka 2, 50-137 Wroclaw (Poland); Kroliczewski, Jaroslaw [Laboratory of Biophysics, Faculty of Biotechnology, University of Wroclaw, 50-148 Wroclaw (Poland); Latos-Grazynski, Lechoslaw [Department of Chemistry, University of Wroclaw, 50-383 Wroclaw (Poland); Olczak, Teresa, E-mail: Teresa.Olczak@biotech.uni.wroc.pl [Laboratory of Biochemistry, Faculty of Biotechnology, University of Wroclaw, Tamka 2, 50-137 Wroclaw (Poland)

    2009-05-29

    Porphyromonas gingivalis, a Gram-negative anaerobic bacterium implicated in the development and progression of chronic periodontitis, acquires heme for growth by a novel mechanism composed of HmuY and HmuR proteins. The aim of this study was to characterize the nature of heme binding to HmuY. The protein was expressed, purified and detailed investigations using UV-vis absorption, CD, MCD, and {sup 1}H NMR spectroscopy were carried out. Ferric heme bound to HmuY may be reduced by sodium dithionite and re-oxidized by potassium ferricyanide. Heme complexed to HmuY, with a midpoint potential of 136 mV, is in a low-spin Fe(III) hexa-coordinate environment. Analysis of heme binding to several single and double HmuY mutants with the methionine, histidine, cysteine, or tyrosine residues replaced by an alanine residue identified histidines 134 and 166 as potential heme ligands.

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

    DEFF Research Database (Denmark)

    Skottrup, Peter Durand; Leonard, Paul

    2011-01-01

    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.

  10. Characterization of a Tn4351-generated hemin uptake mutant of Porphyromonas gingivalis: evidence for the coordinate regulation of virulence factors by hemin.

    OpenAIRE

    Genco, C. A.; Simpson, W.; Forng, R Y; Egal, M; Odusanya, B M

    1995-01-01

    The ability of Porphyromonas gingivalis to acquire iron in the iron-limited environment of the host is crucial to the colonization of this organism. We report here on the isolation and characterization of a transpositional insertion mutant of P. gingivalis A7436 (designated MSM-3) which is defective in the utilization and transport of hemin. P. gingivalis MSM-3 was selected on the basis of its nonpigmented phenotype on anaerobic blood agar following mutagenesis with the Bacteroides fragilis t...

  11. Porphyromonas gingivalis Periodontal Infection and Its Putative Links with Alzheimer's Disease

    Science.gov (United States)

    Singhrao, Sim K.; Harding, Alice; Poole, Sophie; Kesavalu, Lakshmyya; Crean, StJohn

    2015-01-01

    Periodontal disease (PD) and Alzheimer's disease (AD) are inflammatory conditions affecting the global adult population. In the pathogenesis of PD, subgingival complex bacterial biofilm induces inflammation that leads to connective tissue degradation and alveolar bone resorption around the teeth. In health, junctional epithelium seals the gingiva to the tooth enamel, thus preventing bacteria from entering the gingivae. Chronic PD involves major pathogens (Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia) which have an immune armoury that can circumvent host's immune surveillance to create and maintain an inflammatory mediator rich and toxic environment to grow and survive. The neurodegenerative condition, AD, is characterised by poor memory and specific hallmark proteins; periodontal pathogens are increasingly being linked with this dementing condition. It is therefore becoming important to understand associations of periodontitis with relevance to late-onset AD. The aim of this review is to discuss the relevance of finding the keystone periodontal pathogen P. gingivalis in AD brains and its plausible contribution to the aetiological hypothesis of this dementing condition.

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

    Directory of Open Access Journals (Sweden)

    Tribble Gena D

    2009-05-01

    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.

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

    DEFF Research Database (Denmark)

    Pyrc, Krzysztof; Milewska, Aleksandra

    2013-01-01

    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.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2012-10-25

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

  15. The nucleoid-associated protein HU? affects global gene expression in Porphyromonas gingivalis.

    Science.gov (United States)

    Priyadarshini, Richa; Cugini, Carla; Arndt, Annette; Chen, Tsute; Tjokro, Natalia O; Goodman, Steven D; Davey, Mary E

    2013-02-01

    HU is a non-sequence-specific DNA-binding protein and one of the most abundant nucleoid-associated proteins in the bacterial cell. Like Escherichia coli, the genome of Porphyromonas gingivalis is predicted to encode both the HU? (PG1258) and the HU? (PG0121) subunit. We have previously reported that PG0121 encodes a non-specific DNA-binding protein and that PG0121 is co-transcribed with the K-antigen capsule synthesis operon. We also reported that deletion of PG0121 resulted in downregulation of capsule operon expression and produced a P. gingivalis strain that is phenotypically deficient in surface polysaccharide production. Here, we show through complementation experiments in an E. coli MG1655 hupAB double mutant strain that PG0121 encodes a functional HU homologue. Microarray and quantitative RT-PCR analysis were used to further investigate global transcriptional regulation by HU? using comparative expression profiling of the PG0121 (HU?) mutant strain to the parent strain, W83. Our analysis determined that expression of genes encoding proteins involved in a variety of biological functions, including iron acquisition, cell division and translation, as well as a number of predicted nucleoid associated proteins were altered in the PG0121 mutant. Phenotypic and quantitative real-time-PCR (qRT-PCR) analyses determined that under iron-limiting growth conditions, cell division and viability were defective in the PG0121 mutant. Collectively, our studies show that PG0121 does indeed encode a functional HU homologue, and HU? has global regulatory functions in P. gingivalis; it affects not only production of capsular polysaccharides but also expression of genes involved in basic functions, such as cell wall synthesis, cell division and iron uptake. PMID:23175503

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

    OpenAIRE

    Mrudula Patel; Roxanne Naidoo; Foluso John Owotade

    2013-01-01

    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...

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

    OpenAIRE

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

    2013-01-01

    Background: Biofilms are organized communities of microorganisms embedded in a self-produced extracellular polymeric matrix (EPM), often with great phylogenetic variety. Bacteria in the subgingival biofilm are key factors that cause periodontal diseases; among these are the Gram-negative bacteria Fusobacterium nucleatum and Porphyromonas gingivalis. The objectives of this study were to characterize the major components of the EPM and to test the effect of deoxyribonuclease I (DNase I) and pro...

  18. Verification of a topology model of PorT as an integral outer membrane protein in Porphyromonas gingivalis

    OpenAIRE

    Nguyen, Ky-Anh; ?ylicz, Jasiek; Szczesny, Pawel; Sroka, Aneta; Hunter, Neil; Potempa, Jan

    2009-01-01

    PorT is a membrane-associated protein shown to be essential for the maturation and secretion of a class of cysteine proteinases, the gingipains, from the periodontal pathogen Porphyromonas gingivalis. It was previously reported that PorT is located on the periplasmic surface of the inner membrane to function as a chaperone for the maturing proteinases. Our modeling suggested it to be an integral outer membrane protein with eight anti-parallel, membrane-traversing ?-strands. In this report, th...

  19. Identification and Cloning of Genes from Porphyromonas gingivalis after Mutagenesis with a Modified Tn4400 Transposon from Bacteroides fragilis

    OpenAIRE

    Chen, Tsute; Dong, Hong; Tang, Yixin P.; Dallas, Mary M.; Malamy, Michael H.; Duncan, Margaret J.

    2000-01-01

    Porphyromonas gingivalis is a gram-negative, black-pigmented, oral anaerobe strongly associated with adult periodontitis. Previous transposon mutagenesis studies with this organism were based on the Bacteroides transposon Tn4351. Characterization of Tn4351-disrupted genes by cloning has not been an efficient way to analyze large numbers of mutants and is further complicated by the high rate of cointegration of the suicide delivery vector containing Tn4351. In this study, we mutagenized P. gin...

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

    OpenAIRE

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

    2014-01-01

    Histatins are human salivary gland peptides with anti-microbial and anti-inflammatory activities. In this study, we hypothesized that histatin 5 binds to Porphyromonas gingivalis hemagglutinin B (HagB) and attenuates HagB-induced chemokine responses in human myeloid dendritic cells. Histatin 5 bound to immobilized HagB in a surface plasmon resonance (SPR) spectroscopy-based biosensor system. SPR spectroscopy kinetic and equilibrium analyses, protein microarray studies, and I-TASSER structural...

  1. Neolignanos de Krameria ramosissima (A. Gray) S. Watson con actividad contra Porphyromonas gingivalis, evaluación citotóxica y mutagénica / Neolignans from Krameria ramosissima (A. Gray) S. Watson with activity against Porphyromonas gingivalis, cytotoxical and mutagenic evaluations

    Scientific Electronic Library Online (English)

    Laura E., Villarreal-García; Azucena, Oranday-Cárdenas; Myriam A. de la, Garza-Ramos; Catalina, Rivas-Morales; M. Julia, Verde-Star; J. Alberto, Gómez-Treviño; Víctor, Torres-de la Cruz.

    2014-06-01

    Full Text Available Porphyromonas gingivalis, es una de las bacterias asociadas a la enfermedad periodontal, y ha sido relacionada con lesiones coronarias, neumonía y preeclamsia. El propósito de este estudio fue evaluar el extracto metanólico de raíces de Krameria ramosissima contra P. gingivalis (ATCC 53978), determi [...] nar su actividad citotóxica en fibroblastos humanos (ATCC CRL-7222 Hs 274.T) y su potencial mutagénico mediante la prueba de Ames. Las concentraciones a evaluar fueron 500, 400, 300, 200, 150, 100, 75 y 50 µg/mL, siendo la concentración mínima inhibitoria de 300 µg/mL. Mediante cromatografía en columna se obtuvieron 14 fracciones, de las cuales la 7 y la 9 presentaron mayor actividad (P Abstract in english Porphyromonas gingivalis, is one of the bacteria associated with periodontal disease that has been related to coronary artery disease, pneumonia, and preeclampsia. The purpose of this study was to evaluate the methanol extract of roots of Krameria ramosissima against P. gingivalis (ATCC 53978), its [...] cytotoxic activity in human fibroblasts (ATCC CRL-7222 274.T Hs) and its mutagenic potential using the Ames test. Asessed concentrations were 500, 400, 300, 200, 150, 100, 75 and 50 µg/mL, with the minimum inhibitory concentration being of 300 µg/mL. By column chromatography were obtained 14 fractions of which the fractions 7 and 9 showed higher inhibitory activity (P

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

    Science.gov (United States)

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

    2014-12-01

    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

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

    Directory of Open Access Journals (Sweden)

    Miyazawa Haruna

    2012-09-01

    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.

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

    Science.gov (United States)

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

    2013-01-01

    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

  5. HU Protein Affects Transcription of Surface Polysaccharide Synthesis Genes in Porphyromonas gingivalis ? †

    Science.gov (United States)

    Alberti-Segui, Christine; Arndt, Annette; Cugini, Carla; Priyadarshini, Richa; Davey, Mary E.

    2010-01-01

    K-antigen capsule synthesis is an important virulence determinant of the oral anaerobe Porphyromonas gingivalis. We previously reported that the locus required for synthesis of this surface polysaccharide in strain W83 (TIGR identification PG0106 to PG0120) is transcribed as a large (?16.7-kb) polycistronic message. Through sequence analysis, we have now identified a 77-bp inverted repeat located upstream (206 bp) of the start codon of PG0106 that is capable of forming a large hairpin structure. Further sequence analysis just upstream and downstream of the capsule synthesis genes revealed the presence of two genes oriented in the same direction as the operon that are predicted to encode DNA binding proteins: PG0104, which is highly similar (57%) to DNA topoisomerase III, and PG0121, which has high similarity (72%) to DNA binding protein HU (?-subunit). In this report, we show that these two genes, as well as the 77-bp inverted repeat region, are cotranscribed with the capsule synthesis genes, resulting in a large transcript that is ?19.4 kb (based on annotation). We also show that a PG0121 recombinant protein is a nonspecific DNA binding protein with strong affinity to the hairpin structure, in vitro, and that transcript levels of the capsule synthesis genes are downregulated in a PG0121 deletion mutant. Furthermore, we show that this decrease in transcript levels corresponds to a decrease in the amount of polysaccharide produced. Interestingly, expression analysis of another polysaccharide synthesis locus (PG1136 to PG1143) encoding genes involved in synthesis of a surface-associated phosphorylated branched mannan (APS) indicated that this locus is also downregulated in the PG0121 mutant. Altogether our data indicate that HU protein modulates expression of surface polysaccharides in P. gingivalis strain W83. PMID:20889748

  6. HU protein affects transcription of surface polysaccharide synthesis genes in Porphyromonas gingivalis.

    Science.gov (United States)

    Alberti-Segui, Christine; Arndt, Annette; Cugini, Carla; Priyadarshini, Richa; Davey, Mary E

    2010-12-01

    K-antigen capsule synthesis is an important virulence determinant of the oral anaerobe Porphyromonas gingivalis. We previously reported that the locus required for synthesis of this surface polysaccharide in strain W83 (TIGR identification PG0106 to PG0120) is transcribed as a large (?16.7-kb) polycistronic message. Through sequence analysis, we have now identified a 77-bp inverted repeat located upstream (206 bp) of the start codon of PG0106 that is capable of forming a large hairpin structure. Further sequence analysis just upstream and downstream of the capsule synthesis genes revealed the presence of two genes oriented in the same direction as the operon that are predicted to encode DNA binding proteins: PG0104, which is highly similar (57%) to DNA topoisomerase III, and PG0121, which has high similarity (72%) to DNA binding protein HU (?-subunit). In this report, we show that these two genes, as well as the 77-bp inverted repeat region, are cotranscribed with the capsule synthesis genes, resulting in a large transcript that is ?19.4 kb (based on annotation). We also show that a PG0121 recombinant protein is a nonspecific DNA binding protein with strong affinity to the hairpin structure, in vitro, and that transcript levels of the capsule synthesis genes are downregulated in a PG0121 deletion mutant. Furthermore, we show that this decrease in transcript levels corresponds to a decrease in the amount of polysaccharide produced. Interestingly, expression analysis of another polysaccharide synthesis locus (PG1136 to PG1143) encoding genes involved in synthesis of a surface-associated phosphorylated branched mannan (APS) indicated that this locus is also downregulated in the PG0121 mutant. Altogether our data indicate that HU protein modulates expression of surface polysaccharides in P. gingivalis strain W83. PMID:20889748

  7. The GroEL protein of Porphyromonas gingivalis accelerates tumor growth by enhancing endothelial progenitor cell function and neovascularization.

    Science.gov (United States)

    Lin, F-Y; Huang, C-Y; Lu, H-Y; Shih, C-M; Tsao, N-W; Shyue, S-K; Lin, C-Y; Chang, Y-J; Tsai, C-S; Lin, Y-W; Lin, S-J

    2015-06-01

    Porphyromonas gingivalis is a bacterial species that causes destruction of periodontal tissues. Additionally, previous evidence indicates that GroEL from P. gingivalis may possess biological activities involved in systemic inflammation, especially inflammation involved in the progression of periodontal diseases. The literature has established a relationship between periodontal disease and cancer. However, it is unclear whether P. gingivalis GroEL enhances tumor growth. Here, we investigated the effects of P. gingivalis GroEL on neovasculogenesis in C26 carcinoma cell-carrying BALB/c mice and chick eggs in vivo as well as its effect on human endothelial progenitor cells (EPC) in vitro. We found that GroEL treatment accelerated tumor growth (tumor volume and weight) and increased the mortality rate in C26 cell-carrying BALB/c mice. GroEL promoted neovasculogenesis in chicken embryonic allantois and increased the circulating EPC level in BALB/c mice. Furthermore, GroEL effectively stimulated EPC migration and tube formation and increased E-selectin expression, which is mediated by eNOS production and p38 mitogen-activated protein kinase activation. Additionally, GroEL may enhance resistance against paclitaxel-induced cell cytotoxicity and senescence in EPC. In conclusion, P. gingivalis GroEL may act as a potent virulence factor, contributing to the neovasculogenesis of tumor cells and resulting in accelerated tumor growth. PMID:25220060

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

    Directory of Open Access Journals (Sweden)

    Marwan Mansoor Ali Mohammed

    2013-01-01

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

  9. 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

    Directory of Open Access Journals (Sweden)

    M Navarrete

    2010-04-01

    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.

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

    Directory of Open Access Journals (Sweden)

    Yoneda Masato

    2012-02-01

    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.

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

    OpenAIRE

    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

    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...

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

    OpenAIRE

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

    1998-01-01

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

  13. 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)

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

    2012-04-01

    Full Text Available 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.

  14. Identification and cloning of genes from Porphyromonas gingivalis after mutagenesis with a modified Tn4400 transposon from Bacteroides fragilis.

    Science.gov (United States)

    Chen, T; Dong, H; Tang, Y P; Dallas, M M; Malamy, M H; Duncan, M J

    2000-01-01

    Porphyromonas gingivalis is a gram-negative, black-pigmented, oral anaerobe strongly associated with adult periodontitis. Previous transposon mutagenesis studies with this organism were based on the Bacteroides transposon Tn4351. Characterization of Tn4351-disrupted genes by cloning has not been an efficient way to analyze large numbers of mutants and is further complicated by the high rate of cointegration of the suicide delivery vector containing Tn4351. In this study, we mutagenized P. gingivalis with a modified version of the Bacteroides fragilis transposon Tn4400. Plasmid pYT646B carrying the transposon was mobilized from Escherichia coli to P. gingivalis ATCC 33277 by conjugation. Both normal and inverse transposition frequencies were similar (3 x 10(-8)). However, the inverse transposon (Tn4400') contains a pBR322 replicon and a beta-lactamase gene; thus, the cloning of disrupted genomic DNAs from inverse transposition mutants was easily accomplished after ligation of genomic fragments and transformation into E. coli. Thousands of transconjugants could be obtained in a single mating experiment, and inverse transposition was random as demonstrated by Southern hybridization. By this procedure the disrupted genes from P. gingivalis pleiotropic mutants were quickly cloned, sequenced, and identified. PMID:10603421

  15. Characterization of a Tn4351-generated hemin uptake mutant of Porphyromonas gingivalis: evidence for the coordinate regulation of virulence factors by hemin.

    Science.gov (United States)

    Genco, C A; Simpson, W; Forng, R Y; Egal, M; Odusanya, B M

    1995-07-01

    The ability of Porphyromonas gingivalis to acquire iron in the iron-limited environment of the host is crucial to the colonization of this organism. We report here on the isolation and characterization of a transpositional insertion mutant of P. gingivalis A7436 (designated MSM-3) which is defective in the utilization and transport of hemin. P. gingivalis MSM-3 was selected on the basis of its nonpigmented phenotype on anaerobic blood agar following mutagenesis with the Bacteroides fragilis transposon Tn4351. P. gingivalis MSM-3 grew poorly when supplied with hemin as a sole source of iron; however, growth was observed with hemoglobin or inorganic iron. P. gingivalis MSM-3 grown in either hemin-replete or hemin-depleted conditions bound and transported less [14C]hemin or [59Fe]hemin than did the parent strain. At 4 h, P. gingivalis MSM-3 grown in hemin-replete conditions transported only 10,000 pmol of hemin per mg of protein, or 14% of the amount transported by P. gingivalis A7436. Unlike P. gingivalis A7436, hemin binding and transport by P. gingivalis MSM-3 were not tightly regulated by hemin or iron. Examination of P. gingivalis MSM-3 cultures by electron microscopy revealed an overproduction of membrane vesicles, and determination of the dry weight of purified vesicles indicated that P. gingivalis MSM-3 produced twice as much membrane vesicles as did strain A7436. Extracellular vesicles isolated from P. gingivalis MSM-3 also were found to express increased hemolytic and trypsin-like protease activities compared with the parent strain. When inoculated into subcutaneous chambers implanted in mice, P. gingivalis MSM-3 was highly infectious and more invasive than the parent strain, as indicated by secondary lesion formation and death. Taken together, these results indicate that the decreased transport of hemin by P. gingivalis MSM-3 results in the increased expression of several virulence factors which may be coordinately regulated by hemin. PMID:7790057

  16. Coexistence of Porphyromonas gingivalis, Tannerella forsythia and Treponema denticola in the Red Bacterial Complex in Chronic Periodontitis Subjects / Coexistencia de Porphyromonas gingivalis, Tannerella forsythia y Treponema denticola en el Complejo Rojo Bacteriano en Sujetos con Periodontitis Crónica

    Scientific Electronic Library Online (English)

    Carlos Martín, Ardila Medina; Astrid Adriana, Ariza Garcés; Isabel Cristina, Guzmán Zuluaga.

    2014-12-01

    Full Text Available Reportes previos mostraron que la periodontitis se asocia con diferentes microorganismos en lugar de periodontopatógenos particulares en la biopelícula dental. El objetivo del presente estudio fue evaluar la coexistencia y relación entre Porphyromonas gingivalis, Tanerella forsythia y Treponema dent [...] icola en el complejo rojo, señalando su vinculación con la severidad de la periodontitis. En este estudio transversal, 96 sujetos de 33 a 82 años (con 18 dientes residuales) con periodontitis crónica que asistieron a las clínicas dentales de la Universidad de Antioquia en Medellín, Colombia fueron invitados a participar. Se registraron la presencia o ausencia de sangrado al sondaje y placa. La profundidad de sondaje y nivel de inserción clínica se midieron en todas las superficies proximales, bucal y lingual. El muestreo microbiano en pacientes con periodontitis se realizó en los bolsillos mayores a 5 mm. La presencia de P. gingivalis, T. forsythia, y T. denticola se detectó por PCR usando las bolsas periodontales diseñadas para dirigirse a las respectivas secuencias de genes 16S RNAr. La coexistencia de los tres periodontopatógenos fue la más frecuente (25 sujetos). Se observó una asociación estadísticamente significativa entre las tres bacterias (P. gingivalis y T. forsythia, P Abstract in english Previous reports showed that periodontitis is associated with different microorganisms rather than individual periodontopathogens in the dental biofilm. The purpose of the current study was to evaluate the coexistence and relationship among Porphyromonas gingivalis, Tanerella forsythia, and Treponem [...] a denticola in the red complex, noting its association with the severity of periodontitis. In this cross sectional study, 96 subjects, aged 33 to 82 years (with 18 residual teeth) with chronic periodontitis who attended the dental clinics of the Universidad de Antioquia in Medellín, Colombia were invited to participate. The presence or absence of bleeding on probing and plaque were registered. Probing depth and clinical attachment level were measured at all approximal, buccal and lingual surfaces. Microbial sampling on periodontitis patients was performed on pockets >5 mm. The presence of P. gingivalis, T. forsythia, and T. denticola was detected by PCR using primers designed to target the respective 16S rRNA gene sequences. The coexistence of the three periodontopathogens was the most frequent (25 subjects). A statistically significant association between the three bacteria was observed (P. gingivalis and T. forsythia, P

  17. Identification of Dipeptidyl-Peptidase (DPP)5 and DPP7 in Porphyromonas endodontalis, Distinct from Those in Porphyromonas gingivalis

    OpenAIRE

    Nishimata, Haruka; Ohara-nemoto, Yuko; Baba, Tomomi T.; Hoshino, Tomonori; Fujiwara, Taku; Shimoyama, Yu; Kimura, Shigenobu; Nemoto, Takayuki K.

    2014-01-01

    Dipeptidyl peptidases (DPPs) that liberate dipeptides from the N-terminal end of oligopeptides are crucial for the growth of Porphyromonas species, anaerobic asaccharolytic gram negative rods that utilize amino acids as energy sources. Porphyromonas endodontalis is a causative agent of periapical lesions with acute symptoms and Asp/Glu-specific DPP11 has been solely characterized in this organism. In this study, we identified and characterized two P. endodontalis DPPs, DPP5 and DPP7. Cell-ass...

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

    Science.gov (United States)

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

    2014-08-01

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

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

    DEFF Research Database (Denmark)

    Borch, Tanja SkuldbØl; Pedersen, Morten LØbner

    2009-01-01

    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.

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

    Science.gov (United States)

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

    2013-01-01

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

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

    Directory of Open Access Journals (Sweden)

    Lafaurie Gloria

    2008-12-01

    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.

  2. Occurrence of porphyromonas gingivalis and its antibacterial susceptibility to metronidazole and tetracycline in patients with chronic periodontitis

    Scientific Electronic Library Online (English)

    Fredy, Gamboa; Adriana, Acosta; Dabeiba-Adriana, García; Juliana, Velosa; Natalia, Araya; Roberto, Ledergerber.

    2014-12-01

    Full Text Available La periodontitis cronica es una enfermedad infecciosa multifactorial asociada a bacilos Gram-negativos anaerobicos estrictos que se encuentran inmersos en la biopelicula subgingival. Porphyromonas gingivalis, importante patogeno periodontal, es fre - cuentemente detectado en pacientes con periodonti [...] tis cronica. Los aislamientos clinicos de P. gingivalis tienden a ser susceptibles a la mayoria de agentes antimicrobianos; sin embargo, se tiene poca informacion sobre la susceptibilidad antimicrobiana in vitro. El objetivo de este estudio fue determinar la frecuencia de P. gingivalis en pacientes con periodontitis cronica y determinar la susceptibilidad antimicrobiana en terminos de concentracion inhibitoria minima (CIM) de los aislamientos clinicos a metronidazol y tetraciclina. Se realizo un estudio observacional descriptivo en el que se incluyeron 87 pacientes con periodontitis cronica. Las muestras tomadas con conos de papel de la bolsa periodontal se depositaron en caldo tioglicolato, se incubaron durante 4 horas a 37 oC en anaerobiosis y se resembraron en agar anaerobico Wilkins- Chalgren (Oxoid). La identitficacion de los aislamientos se realizo con el sistema RapIDTM ANA II (Remel) y la susceptibilidad antibiotica para metronidazol y tetraciclina se evaluo mediante la tecnica M.I.C.Evaluator (M.I.C.E., Oxoid). En 30 de los 87 pacientes con periodontitis cronica se identifico P. gingivalis, lo que representa una frecuencia de 34.5%. Todos los 30 aislamientos (100%) fueron sensibles al metronidazol con valores de CIM desde 0.015 hasta 4 ug/ml. En cuanto a tetraciclina, 27 aislamientos (90%) fueron sensibles con valores de CIM desde Abstract in english Chronic periodontitis is a multifactorial infectious disease associated with Gram-negative strict anaerobes which are immersed in the subgingival biofilm. Porphyromonas gingivalis, an important periodontal pathogen, is frequently detected in patients with chronic periodontitis. Although isolates of [...] P. gingivalis tend to be susceptible to most antimicrobial agents, relatively little information is available on its in vitro antimicrobial susceptibility. The aim of this study was to determine the frequency of P. gingivalis in patients with chronic periodontitis and to assess antimicrobial susceptibility in terms of minimum inhibitory concentration (MIC) of clinical isolates to metronidazole and tetracycline. A descriptive, observational study was performed including 87 patients with chronic periodontitis. Samples were taken from the periodontal pocket using paper points, which were placed in thioglycollate broth. Samples were incubated for 4 hours at 37°C in anaerobic conditions and finally replated on Wilkins-Chalgren anaerobic agar (Oxoid). Bacteria were identified using the RapIDTMANAII system (Remel) and antimicrobial susceptibility was determined with the M.I.C. Evaluator test (MICE, Oxoid). P. gingivalis was identified in 30 of the 87 patients with chronic periodontitis, which represents a frequency of 34.5%. All 30 isolates (100%) were sensitive to metronidazole, with MIC values ranging from 0015-4ug/ml. Regarding tetracycline, 27 isolates (90%) were sensitive, with MIC values ranging from

  3. The survival rate of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Bacteroides forsythus following 4 randomized treatment modalities.

    Science.gov (United States)

    Shiloah, J; Patters, M R; Dean, J W; Bland, P; Toledo, G

    1997-08-01

    The overall goal of this clinical study was to determine the short-term anti-infective effects of four randomized treatment modalities on Actinobacillus actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), and Bacteroides forsythus (Bf) and determine the effects of bacterial survival on treatment outcomes in patients with adult periodontitis. Twelve adult patients requiring therapy for moderate periodontitis were selected for this study. All patients had at least one tooth in each quadrant that had an inflamed pocket of probing depth > or =5 mm with probing attachment loss that harbored at least one of the following three periodontal pathogens: Aa, Pg, or Bf. The number of target organisms per site was determined pre-operatively, at 1 week, and 1 month and 3 months postoperatively utilizing DNA probes. One quadrant in each patient was randomly assigned to each one of the following four treatment groups: 1) scaling and root planing (SRP group); 2) pocket reduction through osseous surgery and apically-positioned flap (OS group); 3) modified Widman flap (MWF group); and 4) modified Widman flap and topical application of saturated citric acid at pH 1 for 3 minutes (CA group). The 4 treatment modalities were performed in one appointment. No postoperative antibiotics were used. Patients were instructed to supplement their daily oral hygiene with chlorohexidine oral rinse during the study. The results of this investigation indicated that: 1) none of the treatment modalities was effective in eliminating the target species; 2) the incidence of infected sites for all groups was 100% preoperatively; 62.5%, 33.3%, and 31.3% at 1 week, and 1 and 3 months postoperatively, respectively; 3) these infected sites lost 1.1 +/- 0.4 mm of probing attachment compared to gain of 0.0 +/- 0.3 mm for uninfected sites; 4) the infected sites had higher plaque and bleeding on probing 0.9 +/- 0.3, 73 +/- 12%, respectively, compared to 0.3 +/- 0.1 and 30 +/- 8% for the uninfected sites; and 5) no statistically significant differences were detected among the infected sites in regard to gingival index (1.0 +/- 0.2 vs. 0.8 +/- 0.1) or probing depth (3.5 +/- 0.4 vs. 3.0 +/- 0.1 mm). These results indicate that bacterial survival negatively affects the short-term clinical outcomes of non-surgical and surgical periodontal therapy. PMID:9287061

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

    DEFF Research Database (Denmark)

    Borch, T S; Holmstrup, Palle

    2010-01-01

    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.

  5. 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

    OpenAIRE

    Eduardo Augusto Pfau; Mario Julio Avila-Campos

    2005-01-01

    The colonization and antimicrobial susceptibility of P. intermedia and P. gingivalis isolated from peri-implant and gingival sulcus samples were determined. Samples were collected from 30 patients submitted to implant in three different times: at the moment of the surgery, 20 and 60 days after the implant installation. Organisms were identified by using a biochemical tests or API 32-A kit and by PCR. The antimicrobial susceptibility was determined by using an agar dilution method. Nineteen P....

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

    Scientific Electronic Library Online (English)

    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

    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

  7. 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)

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

    2005-09-01

    Full Text Available 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.

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

    Directory of Open Access Journals (Sweden)

    Eduardo Augusto Pfau

    2005-09-01

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

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

    OpenAIRE

    Agnani, G.; Tricot-Doleux, S.; Houalet, S.; Bonnaure-Mallet, M.

    2003-01-01

    We investigated the target structures of the epithelial cells responsible for the attachment of Porphyromonas gingivalis by immunocytofluorimetry, enzyme-linked immunosorbent assay, and confocal microscopy. Integrins (?1, ?3, and ?V) and E-cadherin played no significant role. Carbohydrates (such as ?-d-methylglucoside, l-fucose, d- and l-mannose, N-acetylglucosamine, and N-acetylgalactosamine) had little inhibitory effect on bacterial binding. Enzymatic treatments of the epithelial membranes ...

  10. Evidence and a novel hypothesis for the role of dendritic cells and Porphyromonas gingivalis in adult periodontitis.

    Science.gov (United States)

    Cutler, C W; Jotwani, R; Palucka, K A; Davoust, J; Bell, D; Banchereau, J

    1999-10-01

    We have proposed a novel overall hypothesis and approach to understanding the pathophysiology of adult periodontitis, one of the most common diseases that afflicts the US population. While mortality of the dentition is the most familiar outcome of adult periodontitis, its links with other more severe diseases, including coronary artery disease, respiratory diseases and pre-term labor, cannot be ignored. We have called attention to the many intriguing parallels between adult periodontitis and contact hypersensitivity (CHS). CHS is among the most common of dermatoses that afflicts mankind and one of the most intensively studied of in vivo immune responses. Both adult periodontitis and CHS target the host integument (gingiva or skin) and appear to involve the activation and sensitization of similar subsets of antigen capture and presenting cells, the dendritic cells (DCs), as well as similar T cell subsets. DCs have been termed "nature's adjuvant", being more efficient at antigen-presentation than macrophages or B cells and the only antigen-presenting cells that can stimulate naïve T cells to proliferate. This immunostimulatory capacity can also have detrimental effects for the host, as typified by graft-vs.-host disease and CHS responses. Both AP and CHS involve a predominantly destructive T cell response mediated by both regulatory and effector T cells. In the present paper, we show intriguing evidence that Porphyromonas gingivalis is a unique pathogen in this regard, able to infect, sensitize and activate DCs in vitro and, probably, in situ. Many questions about the role of P. gingivalis-sensitized DCs in adult periodontitis, and of the parallels between adult periodontitis and CHS, however, remain to be answered. PMID:10685369

  11. Structure and mechanism of cysteine peptidase gingipain K (Kgp), a major virulence factor of Porphyromonas gingivalis in periodontitis.

    Science.gov (United States)

    de Diego, Iñaki; Veillard, Florian; Sztukowska, Maryta N; Guevara, Tibisay; Potempa, Barbara; Pomowski, Anja; Huntington, James A; Potempa, Jan; Gomis-Rüth, F Xavier

    2014-11-14

    Cysteine peptidases are key proteolytic virulence factors of the periodontopathogen Porphyromonas gingivalis, which causes chronic periodontitis, the most prevalent dysbiosis-driven disease in humans. Two peptidases, gingipain K (Kgp) and R (RgpA and RgpB), which differ in their selectivity after lysines and arginines, respectively, collectively account for 85% of the extracellular proteolytic activity of P. gingivalis at the site of infection. Therefore, they are promising targets for the design of specific inhibitors. Although the structure of the catalytic domain of RgpB is known, little is known about Kgp, which shares only 27% sequence identity. We report the high resolution crystal structure of a competent fragment of Kgp encompassing the catalytic cysteine peptidase domain and a downstream immunoglobulin superfamily-like domain, which is required for folding and secretion of Kgp in vivo. The structure, which strikingly resembles a tooth, was serendipitously trapped with a fragment of a covalent inhibitor targeting the catalytic cysteine. This provided accurate insight into the active site and suggested that catalysis may require a catalytic triad, Cys(477)-His(444)-Asp(388), rather than the cysteine-histidine dyad normally found in cysteine peptidases. In addition, a 20-Å-long solvent-filled interior channel traverses the molecule and links the bottom of the specificity pocket with the molecular surface opposite the active site cleft. This channel, absent in RgpB, may enhance the plasticity of the enzyme, which would explain the much lower activity in vitro toward comparable specific synthetic substrates. Overall, the present results report the architecture and molecular determinants of the working mechanism of Kgp, including interaction with its substrates. PMID:25266723

  12. Verification of a topology model of PorT as an integral outer membrane protein in Porphyromonas gingivalis

    Science.gov (United States)

    Nguyen, Ky-Anh; ?ylicz, Jasiek; Szczesny, Pawel; Sroka, Aneta; Hunter, Neil; Potempa, Jan

    2009-01-01

    PorT is a membrane-associated protein shown to be essential for the maturation and secretion of a class of cysteine proteinases, the gingipains, from the periodontal pathogen Porphyromonas gingivalis. It was previously reported that PorT is located on the periplasmic surface of the inner membrane to function as a chaperone for the maturing proteinases. Our modeling suggested it to be an integral outer membrane protein with eight anti-parallel, membrane-traversing ?-strands. In this report, the outer membrane localization model was confirmed by the structural and functional tolerance of PorT to hexa-histidine (6×His) tag insertions at selected locations within the protein using site-directed mutagenesis. Interestingly, those PorT mutations adversely affecting gingipain secretion enhanced expression of the porT gene but at the same time suppressed the transcription of the gingipain rgpB gene. Further, PorT mutants deficient in gingipain activities produced significantly more di- and tri-aminopeptidase activities. PorT homologues have been found in restricted members of the Bacteroidetes phylum where there is potential for PorT to participate in the maturation and secretion of proteins with characteristic C-terminal domains (CTD). Knowledge of the cellular localisation of PorT will enable analysis of the role of this protein in a new secretory pathway for the export of gingipains and other CTD-class proteins. PMID:19202082

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

    Science.gov (United States)

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

    2014-01-01

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

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

    Directory of Open Access Journals (Sweden)

    George Ashwin

    2010-01-01

    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.

  15. Porphyromonas gingivalis B cell Antigen Epitope Vaccine, pIRES-ragB'-mGITRL, Promoted RagB-Specific Antibody Production and Tfh Cells Expansion.

    Science.gov (United States)

    Kong, F; Zheng, D; She, P; Ni, P; Zhu, H; Xu, H; Su, Z

    2015-06-01

    The outer membrane protein RagB is one of the major virulence factors of Porphyromonas gingivalis (P. gingivalis). To prevent periodontitis and associated systemic diseases induced by P. gingivalis, we built B cell antigen epitope vaccine characterized by pIRES-ragB'-mGITRL to induce a protective immune responses. The B cell antigen epitope and scrambled peptide of ragB were predicted, cloned into pIRES and constructed pIRES-ragB', pIRES-scrambled epitopes and pIRES-ragB'-mGITRL. pIRES-ragB'-mGITRL was transfected into COS-7 cells. Subsequently, the 6-week-old female BALB/c mice were challenged by P. gingivalis following three time immunization by pIRES, pIRES-ragB', pIRES-scrambled epitopes and pIRES-ragB'-mGITRL. The levels of RagB-specific antibody in the serum and Tfh cells in the spleen were measured by ELISA and flow cytometry, respectively. And higher levels of RagB-specific IgG were produced in the immunized mice with pIRES-ragB'-mGITRL. Additionally, the number of Tfh cells was also expanded and lesions were diminished in pIRES-ragB'-mGITRL mice comparing with control groups. Our results clearly demonstrated that P. gingivalis B cell antigen epitope vaccine, pIRES-ragB'-mGITRL, could induce protective immune responses. Furthermore, our data also indicated that pIRES-ragB'-mGITRL was a potential therapeutic vaccine against P. gingivalis. PMID:25689343

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

    Directory of Open Access Journals (Sweden)

    Jayanta Kumar Patra

    2015-04-01

    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.

  17. Bactericidal effect of extracts and metabolites of Robinia pseudoacacia L. on Streptococcus mutans and Porphyromonas gingivalis causing dental plaque and periodontal inflammatory diseases.

    Science.gov (United States)

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

    2015-01-01

    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

  18. Analytical performance of an immunologic-based periodontal bacterial test for simultaneous detection and differentiation of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Prevotella intermedia.

    Science.gov (United States)

    Snyder, B; Ryerson, C C; Corona, H; Grogan, E A; Reynolds, H S; Contestable, P B; Boyer, B P; Mayer, J; Mangan, T; Norkus, N; Zambon, J J; Genco, R J

    1996-05-01

    The analytical performance of a membrane-based immunoassay for the simultaneous detection and differentiation of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, and Prevotella intermedia (including Prevotella nigrescens) was investigated. Positive reactions were observed for 71 of 71 reference strains and recent oral isolates of A. actinomycetemcomitans, P. gingivalis, and P. intermedia. No cross-reactivity was observed with 39 other common oral and environmental species. The specificity of the test was unaffected by the presence of potential oral interferents including whole blood, white blood cells, mucin, saliva, toothpastes, and oral rinses. A proficiency test by dental professionals using a standardized set of unknown simulated samples yielded a sensitivity of 97% (116/120) and a 100% specificity (240/ 240). An additional group including dental professionals and high school students was shown to be 99% proficient (1385/1397) in distinguishing proper from improper test function when processing control samples with normal test devices and devices with simulated error conditions. Comparisons to a culture standard for 104 subgingival plaque samples collected from 26 adult periodontitis patients yielded > 98% specificity for each of the test bacteria. In addition, the detection threshold for the test was determined to be equivalent to 10(4) cultivable test bacteria when compared to the culture standard. The data indicate that this membrane immunoassay is a valid and easy-to-use method for the detection of A. actinomycetemcomitans, P. gingivalis, and P. intermedia in subgingival plaque, at levels above the detection threshold of the test. PMID:8724708

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

    Directory of Open Access Journals (Sweden)

    AJ Quintero

    2011-08-01

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

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

    Scientific Electronic Library Online (English)

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

    2011-08-01

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

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

    Directory of Open Access Journals (Sweden)

    Stojanovi? Nikola

    2014-01-01

    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.

  2. 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)

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

    2010-04-01

    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 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.

  3. 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)

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

    2012-12-01

    Full Text Available 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.

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

    Directory of Open Access Journals (Sweden)

    Bronislaw L. Slomiany

    2010-12-01

    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.

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

    OpenAIRE

    L. Gölz; Memmert, S.; Rath-Deschner, B.; A. Jäger; Appel, T.; Baumgarten, G; W. Götz; Frede, S.

    2014-01-01

    Oxidative stress is characterized by an accumulation of reactive oxygen species (ROS) and plays a key role in the progression of inflammatory diseases. We hypothesize that hypoxic and inflammatory events induce oxidative stress in the periodontal ligament (PDL) by activating NOX4. Human primary PDL fibroblasts were stimulated with lipopolysaccharide from Porphyromonas gingivalis (LPS-PG), a periodontal pathogen bacterium under normoxic and hypoxic conditions. By quantitative PCR, immunoblot, ...

  6. 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)

    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

    Full Text Available Objective: The aim of this study was to compare the production of the chemokines CCL3 and CXCL12 by cultured dental pulp fibroblasts from permanent (PDPF) and deciduous (DDPF) teeth under stimulation by Porphyromonas gingivalis LPS (PgLPS). Material and Methods: Primary culture of fibroblasts 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.

  7. A YadA-like autotransporter, Hag1 in Veillonella atypica is a multivalent hemagglutinin involved in adherence to oral streptococci, Porphyromonas gingivalis, and human oral buccal cells.

    Science.gov (United States)

    Zhou, P; Liu, J; Merritt, J; Qi, F

    2015-08-01

    Dental biofilm development is a sequential process, and adherence between microbes and the salivary pellicle (adhesion) as well as among different microbes (co-adhesion or coaggregation) plays a critical role in building a biofilm community. The Veillonella species are among the most predominant species in the oral cavity and coaggregate with many initial, early, middle, and late colonizers. Similar to oral fusobacteria, they are also considered bridging species in biofilm development. However, the mechanism of this ability has yet to be reported, due to the previous lack of a genetic transformation system in the entire genus. In this study, we used our recently discovered transformable Veillonella strain, Veillonella atypica OK5, to probe the mechanism of coaggregation between Veillonella species and other oral bacteria. By insertional inactivation of all eight putative hemagglutinin genes, we identified one gene, hag1, which is involved in V. atypica coaggregation with the initial colonizers Streptococcus gordonii, Streptococcus oralis and Streptococcus cristatus, and the periodontal pathogen Porphyromonas gingivalis. The hag1 mutant also abolished adherence to human buccal cells. Inhibition assays using various chemical or physiological treatments suggest different mechanisms being involved in coaggregation with different partners. The entire hag1 gene was sequenced and shown to be the largest known bacterial hemagglutinin gene. PMID:25440509

  8. Porphyromonas gingivalis Fimbriae Use ?2 Integrin (CD11/CD18) on Mouse Peritoneal Macrophages as a Cellular Receptor, and the CD18 ? Chain Plays a Functional Role in Fimbrial Signaling

    OpenAIRE

    Takeshita, Akira; Murakami, Yukio; Yamashita, Yoshinori; Ishida, Masami; Fujisawa, Seiichiro; Kitano, Shigeo; Hanazawa, Shigemasa

    1998-01-01

    In this study, we demonstrate that Porphyromonas gingivalis fimbriae use molecules of ?2 integrin (CD11/CD18) on mouse peritoneal macrophages as cellular receptors and also show that the ? chain (CD18) may play a functional role in signalling for the fimbria-induced expression of interleukin-1? (IL-1?) and tumor necrosis factor alpha (TNF-?) genes in the cells. Using a binding assay with 125I-labeled fimbriae, we observed that fimbrial binding to the macrophages was inhibited by treatment wit...

  9. Role of Mitogen-Activated Protein Kinases and NF-?B in the Regulation of Proinflammatory and Anti-Inflammatory Cytokines by Porphyromonas gingivalis Hemagglutinin B

    Science.gov (United States)

    Zhang, Ping; Martin, Michael; Michalek, Suzanne M.; Katz, Jannet

    2005-01-01

    Hemagglutinin B (HagB) is a nonfimbrial adhesin expressed on the surface of Porphyromonas gingivalis and has been implicated as a potential virulence factor involved in mediating the attachment of the bacteria to host cells. However, the molecular mechanisms underlying host responses to HagB and their roles in pathogenesis have yet to be elucidated. Mitogen-activated protein kinases (MAPKs) are activated following engagement of a variety of cell surface receptors via dual tyrosine and threonine phosphorylation and are thought to be involved in various cellular responses. The purpose of this study was to determine the role of intracellular signaling pathways including the MAPKs and NF-?B in regulating the production of proinflammatory and anti-inflammatory cytokines following stimulation of murine macrophages with recombinant HagB (rHagB). Stimulation of peritoneal macrophages with rHagB resulted in the production of the proinflammatory cytokines interleukin-12p40 (IL-12p40), gamma interferon (IFN-?), and tumor necrosis factor alpha, as well as the anti-inflammatory cytokine IL-10. We also demonstrated the activation of extracellular signal-related kinase (ERK), c-Jun NH2-terminal protein kinase (JNK), and p38 MAPKs by rHagB-stimulated macrophages. Furthermore, blocking of the ERK and p38 signaling pathways by using specific inhibitors revealed differential regulatory roles in the rHagB-mediated production of proinflammatory and anti-inflammatory cytokines. ERK and p38 were important in down-regulation of IL-12p40 and IFN-? production and up-regulation of IL-10 production. The enhanced levels of IL-12p40 in rHagB-stimulated macrophages by inhibition of ERK or p38 activity were partially attributable to the inhibition of IL-10 production. Moreover, NF-?B was found to be critical for up-regulation of IL-12p40 and down-regulation of IL-10 production in rHagB-stimulated macrophages. Taken together, our results demonstrate a role for the p38 and ERK pathways and the transcription factor NF-?B in modulating key immunoregulatory cytokines involved in the development of immune responses to P. gingivalis HagB. PMID:15972486

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

    DEFF Research Database (Denmark)

    van der Post, Sjoerd; Subramani, Durai B

    2013-01-01

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

  11. The RgpA-Kgp proteinase-adhesin complexes of Porphyromonas gingivalis Inactivate the Th2 cytokines interleukin-4 and interleukin-5.

    Science.gov (United States)

    Tam, Vivian; O'Brien-Simpson, Neil M; Chen, Yu-Yen; Sanderson, Colin J; Kinnear, Beverley; Reynolds, Eric C

    2009-04-01

    The RgpA-Kgp proteinase-adhesin complexes are a primary virulence factor of Porphyromonas gingivalis, a major pathogen in the development of chronic periodontitis. The RgpA-Kgp complexes have been suggested to bias the immune response to a Th2 phenotype in disease by hydrolysis of Th1 cytokines. Here, we show that the RgpA-Kgp complexes hydrolyze and inactivate interleukin-4 (IL-4) and IL-5 under physiologically relevant conditions. Using the IL-4/IL-5-dependent TF1.8 T-cell line, it was found that at equimolar ratios of cytokine to RgpA-Kgp complexes, IL-4 and IL-5 were inactivated in the culture medium. The inactivation of IL-4 and IL-5 was RgpA-Kgp concentration dependent, as at an enzyme-to-cytokine molar ratio of 1:8, the bioactivity of the cytokines was greater than at the higher concentration of RgpA-Kgp of 1:1. Furthermore, inactivation of the cytokines by the RgpA-Kgp complexes was time dependent, as longer preincubation times resulted in lower cytokine activity. IL-5 was found to be slightly more resistant to inactivation than IL-4. Mass spectrometric analyses of IL-4 and IL-5 showed that hydrolysis by RgpA-Kgp complexes was C terminal to Arg and Lys residues of the cytokines. The peptides released indicated that the regions of IL-4 and IL-5 important for bioactivity were being hydrolyzed in the first 15 min of incubation. The ability of the RgpA-Kgp complexes to degrade Th2 cytokines may contribute to immune dysregulation and may play a role in the pathology of chronic periodontitis. PMID:19168731

  12. Extract from Rumex acetosa L. for Prophylaxis of Periodontitis: Inhibition of Bacterial In Vitro Adhesion and of Gingipains of Porphyromonas gingivalis by Epicatechin-3-O-(4??8)-Epicatechin-3-O-Gallate (Procyanidin-B2-Di-Gallate)

    Science.gov (United States)

    Schmuch, Jana; Beckert, Sabine; Brandt, Simone; Löhr, Gesine; Hermann, Fabian; Schmidt, Thomas J.; Beikler, Thomas; Hensel, Andreas

    2015-01-01

    Background The aerial parts of Rumex acetosa L. have been used in traditional European medicine for inflammatory diseases of the mouth epithelial tissue. The following study aimed to investigate the influence of a proanthocyanidin-enriched extract from R. acetosa extract against the adhesion of Porphyromonas gingivalis (P. gingivalis), a pathogen strongly involved in chronic and aggressive periodontitis. A further goal was to define the bioactive lead structures responsible for a potential antiadhesive activity and to characterize the underlying molecular mechanisms of the antiadhesive effects. Methodology An extract of R. acetosa (RA1) with a defined mixture of flavan-3-ols, oligomeric proanthocyanidins and flavonoids, was used. Its impact on P. gingivalis adhesion to KB cells was studied by flow cytometry, confocal laser scanning microscopy and in situ adhesion assay using murine buccal tissue. RA1 and its compounds 1 to 15 were further investigated for additional effects on gingipain activity, hemagglutination and gene expression by RT-PCR. Principal Findings RA1 (5 to 15 ?g/mL) reduced P. gingivalis adhesion in a dose-dependent manner to about 90%. Galloylated proanthocyanidins were confirmed to be responsible for this antiadhesive effect with epicatechin-3-O-gallate-(4?,8)-epicatechin-3’-O-gallate (syn. procyanidin B2-di-gallate) being the lead compound. Ungalloylated flavan-3-ols and oligomeric proanthocyanidins were inactive. RA1 and the galloylated proanthocyanidins strongly interact with the bacterial virulence factor Arg-gingipain, while the corresponding Lys-gingipain was hardly influenced. RA1 inhibited also hemagglutination. In silico docking studies indicated that epicatechin-3-O-gallate-(4?,8)-epicatechin-3’-O-gallate interacts with the active side of Arg-gingipain and hemaglutinin from P. gingivalis; the galloylation of the molecule seems to be responsible for fixation of the ligand to the protein. In conclusion, the proanthocyanidin-enriched extract RA1 and its main active constituent procyanidin B2-di-gallate protect cells from P. gingivalis infection by inhibiting bacterial adhesion to the host cell. RA1 and procyanidin B2-di-gallate appear to be promising candidates for future cytoprotective preparations for oral mouth care products. PMID:25803708

  13. 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)

    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

    Full Text Available 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.

  14. 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)

    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

    Full Text Available 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

  15. Human beta-defensin 3 binds to hemagglutinin B (rHagB), a non-fimbrial adhesin from Porphyromonas gingivalis, and attenuates a pro-inflammatory cytokine response.

    Science.gov (United States)

    Pingel, Lindsey C; Kohlgraf, Karl G; Hansen, Christopher J; Eastman, Christopher G; Dietrich, Deborah E; Burnell, Kindra K; Srikantha, Rupasree N; Xiao, Xiangjun; Bélanger, Myriam; Progulske-Fox, Ann; Cavanaugh, Joseph E; Guthmiller, Janet M; Johnson, Georgia K; Joly, Sophie; Kurago, Zoya B; Dawson, Deborah V; Brogden, Kim A

    2008-01-01

    Regulatory mechanisms in mucosal secretions and tissues recognize antigens and attenuate pro-inflammatory cytokine responses. Here, we asked whether human beta-defensin 3 (HBD3) serves as an upstream suppressor of cytokine signaling that binds and attenuates pro-inflammatory cytokine responses to recombinant hemagglutinin B (rHagB), a non-fimbrial adhesin from Porphyromonas gingivalis strain 381. We found that HBD3 binds to immobilized rHagB and produces a significantly higher resonance unit signal in surface plasmon resonance spectroscopic analysis, than HBD2 and HBD1 that are used as control defensins. Furthermore, we found that HBD3 significantly attenuates (Pdendritic cell culture supernatants and the extracellular signal-regulated kinases (ERK 1/2) response in human myeloid dendritic cell lysates. Thus, HBD3 binds rHagB and this interaction may be an important initial step to attenuate a pro-inflammatory cytokine response and an ERK 1/2 response. PMID:18711400

  16. Hypoxia and P. gingivalis synergistically induce HIF-1 and NF-?B activation in PDL cells and periodontal diseases.

    Science.gov (United States)

    Gölz, L; Memmert, S; Rath-Deschner, B; Jäger, A; Appel, T; Baumgarten, G; Götz, W; Frede, S

    2015-01-01

    Periodontitis is characterized by deep periodontal pockets favoring the proliferation of anaerobic bacteria like Porphyromonas gingivalis (P. gingivalis), a periodontal pathogen frequently observed in patients suffering from periodontal inflammation. Therefore, the aim of the present study was to investigate the signaling pathways activated by lipopolysaccharide (LPS) of P. gingivalis (LPS-PG) and hypoxia in periodontal ligament (PDL) cells. The relevant transcription factors nuclear factor-kappa B (NF-?B) and hypoxia inducible factor-1 (HIF-1) were determined. In addition, we analyzed the expression of interleukin- (IL-) 1?, matrix metalloproteinase-1 (MMP-1), and vascular endothelial growth factor (VEGF) in PDL cells on mRNA and protein level. This was accomplished by immunohistochemistry of healthy and inflamed periodontal tissues. We detected time-dependent additive effects of LPS-PG and hypoxia on NF-?B and HIF-1? activation in PDL cells followed by an upregulation of IL-1?, MMP-1, and VEGF expression. Immunohistochemistry performed on tissue samples of gingivitis and periodontitis displayed an increase of NF-?B, HIF-1, and VEGF immunoreactivity in accordance with disease progression validating the importance of the in vitro results. To conclude, the present study underlines the significance of NF-?B and HIF-1? and their target genes VEGF, IL-1?, and MMP-1 in P. gingivalis and hypoxia induced periodontal inflammatory processes. PMID:25861162

  17. Arginine deiminase inhibits Porphyromonas gingivalis surface attachment

    OpenAIRE

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

    2013-01-01

    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) prod...

  18. Actinobacillus Actinomycetemcomitans y Porphyromonas Gingivales como principales patógenos periodontales

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    A Bascones

    2000-09-01

    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.

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

    Science.gov (United States)

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

    2011-01-01

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

  20. LPS from P. gingivalis and hypoxia increases oxidative stress in periodontal ligament fibroblasts and contributes to periodontitis.

    Science.gov (United States)

    Gölz, L; Memmert, S; Rath-Deschner, B; Jäger, A; Appel, T; Baumgarten, G; Götz, W; Frede, S

    2014-01-01

    Oxidative stress is characterized by an accumulation of reactive oxygen species (ROS) and plays a key role in the progression of inflammatory diseases. We hypothesize that hypoxic and inflammatory events induce oxidative stress in the periodontal ligament (PDL) by activating NOX4. Human primary PDL fibroblasts were stimulated with lipopolysaccharide from Porphyromonas gingivalis (LPS-PG), a periodontal pathogen bacterium under normoxic and hypoxic conditions. By quantitative PCR, immunoblot, immunostaining, and a specific ROS assay we determined the amount of NOX4, ROS, and several redox systems. Healthy and inflamed periodontal tissues were collected to evaluate NOX4 and redox systems by immunohistochemistry. We found significantly increased NOX4 levels after hypoxic or inflammatory stimulation in PDL cells (P < 0.001) which was even more pronounced after combination of the stimuli. This was accompanied by a significant upregulation of ROS and catalase (P < 0.001). However, prolonged incubation with both stimuli induced a reduction of catalase indicating a collapse of the protective machinery favoring ROS increase and the progression of inflammatory oral diseases. Analysis of inflamed tissues confirmed our hypothesis. In conclusion, we demonstrated that the interplay of NOX4 and redox systems is crucial for ROS formation which plays a pivotal role during oral diseases. PMID:25374447

  1. Porphyromonas gingivalis: keeping the pathos out of the biont

    OpenAIRE

    Carla Cugini; Vanja Klepac-Ceraj; Elze Rackaityte; Riggs, James E.; Mary E. Davey

    2013-01-01

    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...

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

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    Yukie Kogawa

    2011-01-01

    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

  3. Lipopolysaccharide Endotoxins

    OpenAIRE

    Raetz, Christian R H; Whitfield, Chris

    2001-01-01

    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-...

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

    Scientific Electronic Library Online (English)

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

    2010-06-01

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

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

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    Stefanon Ivanita

    2011-05-01

    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.

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

    OpenAIRE

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

    2014-01-01

    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...

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

    Directory of Open Access Journals (Sweden)

    José Alexandre de Andrade

    2010-06-01

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

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

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    Ara Toshiaki

    2012-03-01

    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.

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

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    Babak Amoian

    2014-04-01

    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.

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

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    Paknejad M.

    2005-05-01

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

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

    Science.gov (United States)

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

    2014-10-01

    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

  12. Draft Genome Sequences of Porphyromonas crevioricanis JCM 15906T and Porphyromonas cansulci JCM 13913T Isolated from a Canine Oral Cavity

    OpenAIRE

    Sakamoto, Mitsuo; Tanaka, Naoto; Shiwa, Yuh; Yoshikawa, Hirofumi; Ohkuma, Moriya

    2013-01-01

    Here, we report the draft genome sequences of Porphyromonas crevioricanis JCM 15906T and Porphyromonas cansulci JCM 13913T, which were isolated from a canine oral cavity and were recently united under the single species P. crevioricanis. These two genome sequences are very similar, and yet a high degree of genome rearrangements is observed.

  13. Draft Genome Sequences of Porphyromonas crevioricanis JCM 15906T and Porphyromonas cansulci JCM 13913T Isolated from a Canine Oral Cavity

    Science.gov (United States)

    Tanaka, Naoto; Shiwa, Yuh; Yoshikawa, Hirofumi; Ohkuma, Moriya

    2013-01-01

    Here, we report the draft genome sequences of Porphyromonas crevioricanis JCM 15906T and Porphyromonas cansulci JCM 13913T, which were isolated from a canine oral cavity and were recently united under the single species P. crevioricanis. These two genome sequences are very similar, and yet a high degree of genome rearrangements is observed. PMID:23887912

  14. El Lipopolisacárido / Lipopolysaccharide

    Scientific Electronic Library Online (English)

    Stefany, Romero Hurtado; Carlos Arturo, Iregui.

    2010-06-01

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

  15. 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

    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

    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...

  16. 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

  17. Immunochemical characterization of rough Brucella lipopolysaccharides.

    OpenAIRE

    Moreno, E.; Jones, L. M.; Berman, D. T.

    1984-01-01

    Lipopolysaccharides (LPS) were extracted from rough strains of Brucella abortus and Brucella melitensis and from strains of the naturally occurring rough species Brucella ovis and Brucella canis. Brucella rough lipopolysaccharides (R-LPS) were readily distinguished from Brucella smooth lipopolysaccharides (S-LPS) and enterobacterial R-LPS, by their chemical, physical, and serological characteristics. B. ovis R-LPS was differentiated from B. abortus, B. melitensis, and B. canis R-LPS by its re...

  18. 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)

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

    Full Text Available 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.

  19. Draft Genome Sequences of 26 Porphyromonas Strains Isolated from the Canine Oral Microbiome

    Science.gov (United States)

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

    2015-01-01

    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

  20. Serum antibodies to oral Bacteroides asaccharolyticus (Bacteroides gingivalis): relationship to age and periondontal disease.

    OpenAIRE

    Mouton, C; Hammond, P G; Slots, J.; Genco, R J

    1981-01-01

    An enzyme-linked immunosorbent assay microplate method was used for measuring levels of antibody specific for the oral serotype of Bacteroides asaccharolyticus (Bacteroides gingivalis) in serum samples obtained from umbilical cords, infants, children, periodontally normal adults, and edentulous adults. Serum from patients with various periodontal diseases, including adult periodontitis, localized juvenile periodontitis, generalized juvenile periodontitis, post-localized juvenile periodontitis...

  1. 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)

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

    OpenAIRE

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

    1986-01-01

    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...

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

    OpenAIRE

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

    1985-01-01

    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 ...

  4. Purification and characterization of a novel type of fimbriae from the oral anaerobe Bacteroides gingivalis.

    OpenAIRE

    Yoshimura, F.; Takahashi, K.; Nodasaka, Y.; Suzuki, T.

    1984-01-01

    Fimbriae and their constituent protein (fimbrilin) were purified to homogeneity from the bacterial wash fluid and cell lysate fraction, respectively, of Bacteroides gingivalis 381. Fimbriae, observed by negative staining, were curly, single-stranded filaments with a diameter of ca. 5 nm. The apparent molecular weight of the fimbrilin was 43,000. Fimbriae were resistant to sodium dodecyl sulfate denaturation at 70 degrees C. Heating at 100 degrees C in sodium dodecyl sulfate was needed to comp...

  5. Lipopolysaccharide (LPS) stimulation of fungal secondary metabolism

    OpenAIRE

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

    2014-01-01

    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...

  6. Neutralization of bacterial lipopolysaccharides by human plasma.

    OpenAIRE

    Warren, H. S.; Novitsky, T. J.; Ketchum, P. A.; Roslansky, P. F.; Kania, S.; Siber, G. R.

    1985-01-01

    To quantify the neutralization of bacterial lipopolysaccharide (LPS) by human plasma, dilutions of Escherichia coli O113 LPS were incubated with plasma, followed by the addition of Limulus amebocyte lysate (LAL). The reaction between the LPS and LAL was monitored spectrophotometrically, and the concentration of LPS resulting in 50% lysate response (LR50) was determined. Analysis of 145 outdated plasma samples yielded a range of LR50 between 6 and 1,500 ng/ml. Pools of plasma with high and low...

  7. [Phytotoxic properties of Ralstonia solanacearum lipopolysaccharides].

    Science.gov (United States)

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

    2014-01-01

    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

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

    Scientific Electronic Library Online (English)

    Silvio, Favoreto Junior; Maria Inês, Machado.

    1995-12-01

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

  9. Role for moesin in lipopolysaccharide-stimulated signal transduction.

    Science.gov (United States)

    Iontcheva, Iveta; Amar, Salomon; Zawawi, Khalid H; Kantarci, Alpdogan; Van Dyke, Thomas E

    2004-04-01

    Moesin is a 78-kDa protein with diverse functions in linking the cytoskeleton to the membrane while controlling cell shape, adhesion, locomotion, and signaling. The aim of this study was to characterize the expression and localization of moesin in mononuclear phagocytes by using confocal microscopy, flow cytometry, immunoprecipitation, and Western blotting and to analyze the function of moesin as a lipopolysaccharide receptor, utilizing an antisense oligonucleotide approach to knock down the moesin gene. Results revealed that moesin is expressed on the surface of monocytes/macrophages and surface expression is increased after lipopolysaccharide stimulation. The total protein mass of moesin is increased in monocytes after lipopolysaccharide stimulation. Immunoprecipitation showed that moesin coprecipitates with TLR4, a well-known lipopolysaccharide receptor, suggesting an early role of moesin in the formation of the initiation complex for lipopolysaccharide signaling. Two antisense and two control sense oligonucleotides were synthesized and introduced every 4 h for 48 h in adherent macrophage-like cells. Cells were then stimulated with lipopolysaccharide for 4 h, and the supernatants were assayed for tumor necrosis factor alpha (TNF-alpha) production. Cell lysates were assayed for moesin expression by Western blotting immediately after the 48-h treatment period and also after 116 h of recovery to assess the return of moesin expression and function. Moesin gene expression was completely suppressed after 48 h of incubation with antisense oligonucleotides. The antisense elimination of moesin gene expression led to a significant reduction of lipopolysaccharide-induced TNF-alpha secretion. Restoration of moesin gene expression led to restoration of TNF-alpha production. These data suggest an important role for moesin in lipopolysaccharide-induced TNF-alpha production, highlighting its importance in lipopolysaccharide-mediated signal transduction. PMID:15039356

  10. Evaluation of Phototherapy Antimicrobial Activity Against Porphyromonas Gingivales and Prevotella Melaninogenica

    Directory of Open Access Journals (Sweden)

    Ana Maria Gondim VALENÇA

    2006-08-01

    Full Text Available Objective: The objective of the present work went verify, in vitro, the effect antimicrobial of those solutions about two species bacterial associated with disease periodontal. Method: The dyes, besides clorexidine at 0.12 as group controls positive, and alcohol of cereals, used in prepare it of the dyes, as negative control, they were diluted in saline solution of 1:2 up to 1:128. Using the method of the diffusion in agar, the stumps of reference Porphyromonas gingivales ATCC 49417 and Prevotella melaninogenica ATCC 25845, was sowed in half BHI agar enriched with yeast extract (0,5% and incubated anaerobically, to 37th C for 3 days. Results: The results demonstrated that Plantain and Sage possess action antibacterial on the two stumps in test, as well as the clorexidine. Even so the dye of Taheebo didn't interfere in the growth of P. gingivales, being sensitive only P. melaninogenica to this dye. The stumps came resistant to the alcohol of cereals. Conclusion: It is ended that the Plantain dyes and Sage present larger spectrum of performance antibiotics, when compared the dye of Taheebo, being not its effect influenced by the alcohol used in its production.

  11. Lipopolysaccharide Extracellular Emulsifier Produced by Penicillium citrinum

    Directory of Open Access Journals (Sweden)

    M.M. Camargo de Morais

    2006-01-01

    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.

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

    Scientific Electronic Library Online (English)

    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

    Full Text Available 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.

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

    Directory of Open Access Journals (Sweden)

    Sandra Penteado

    2013-06-01

    Full Text Available The present study investigated whether late maternal inflammation disrupts the mother/pup interaction, resulting in longlasting effects on pup behavior and alterations in biological pathways, thereby programming prepubertal behavior and the pups’ inflammatory responses after bacterial endotoxin treatment. Female rats received 100 ?g/kg lipopolysaccharide (LPS or .9% saline solution on gestation day 18. Reproductive performance was observed at birth. On lactation 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.

  14. Lipopolysaccharide (LPS) stimulation of fungal secondary metabolism.

    Science.gov (United States)

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

    2014-07-01

    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

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

    Science.gov (United States)

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

    2015-04-01

    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

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

    Scientific Electronic Library Online (English)

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

    2002-04-01

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

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

    Directory of Open Access Journals (Sweden)

    F. Gallardo

    2002-04-01

    Full Text Available El objetivo de la presente investigación fue estudiar los efectos de un gel de tetraciclina al 5% sobre los niveles de 3 microorganismos asociados al desarrollo de la periodontitis rápidamente progresiva (PRP. En un total de 20 pacientes con PRP se seleccionaron 5 dientes por paciente con bolsas periodontales > a 5 mm. con sangrado al sondaje periodontal en los cuales se efectuaron los siguientes tratamientos: 1. Control. 2. Aplicación de un gel de tetraciclina al 5% (T. 3. Placebo (Pl. 4. Raspado y alisado radicular (RA. 5. T + RA .De forma previa, en cada sitio seleccionado, se tornaron muestras de placa subgingival con un cono de papel estéril para detectar y cuantificar la presencia de P. gingivalis , P. intermedia y A. actinomycetemcomitans, mediante el uso de sondas DNA (OMNIGENE, U.S.A..Se dieron instrucciones de higiene oral, y se efectuó un nuevo control microbiológico a los 60 días El análisis estadístico de los resultados demostró lo siguiente: 1. Ninguno de los tratamientos redujo significativamente los niveles de A. actinomycetemcomitans. 2. Se detectó una reducción significativa de P. gingivalis en los sitios tratados con (R.A. (p The purpose of this investigation was to study the effects of local delivery of a tetracycline 5% gel on the levels of 3 bacteria associated to development of rapidly progressive periodontitis. In a sample of 20 patients, five teeth were selected from each patient with periodontal pockets > 5 mm and bleeding upon probing. One of the following treatments were done at each selected site: 1. Control (no treatment. 2. Local delivery of a 5% tetracycline gel. 3. Placebo gel. 4. Scaling and root planing. 5. Scaling and root planing + local application of tetracycline gel. Previously, at each selected site samples of subgingival plaque were taken with sterile paper points in order to detecte and quantify the presence of P.gingivalis, P.intermedia and A. actinomycetemcomitans, by using DNA probe technology (OMNIGENE, U.S.A.. Oral hygiene instructions were given to each patient and new samples of subgingival plaque were obtained at 60 days. Statistical analysis of results showed the following 1. No significant reductions of A. actinomycetemcomitans were found with performed treatments. 2. Scaling and root planing reduced the levels of P. gingivalis (p 0.02 or when the later was combined with tetracycline (p <0.05. 3. All treatments significantly reduced levels of P.intermedia. Although bacteria studied has shown sensitivity to tetracyclines and despite high levels of antibiotic that are obtained following its local delivery in a gel, the reduced microbiologic effects that were seen could be ascribed to rapid removal of gel by the gingival crevicular fluid from studied sites.

  18. Genetic derepression of a developmentally regulated lipopolysaccharide antigen from Rhizobium leguminosarum 3841.

    OpenAIRE

    Wood, E. A.; Butcher, G. W.; Brewin, N. J.; Kannenberg, E. L.

    1989-01-01

    Monoclonal antibody AFRC MAC 203 recognizes a developmentally regulated lipopolysaccharide antigen in Rhizobium leguminosarum bv. viciae 3841. Transposon-induced mutants that constitutively expressed MAC 203 antigen were isolated. These strains were morphologically normal, showed no gross abnormalities in lipopolysaccharide size distribution on sodium dodecyl sulfate-polyacrylamide gels, and induced normal nitrogen-fixing nodules. However, the mutants lacked lipopolysaccharide epitopes recogn...

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

  20. Correlation between Either Cupriavidus or Porphyromonas and Primary Pulmonary Tuberculosis Found by Analysing the Microbiota in Patients’ Bronchoalveolar Lavage Fluid

    Science.gov (United States)

    Zhou, Yuhua; Lin, Feishen; Cui, Zelin; Zhang, Xiangrong; Hu, Chunmei; Shen, Tian; Chen, Chunyan; Zhang, Xia; Guo, Xiaokui

    2015-01-01

    Pulmonary tuberculosis (TB) has gained attention in recent decades because of its rising incidence trend; simultaneously, increasing numbers of studies have identified the relationship between microbiota and chronic infectious diseases. In our work, we enrolled 32 patients with primary TB characterised by unilateral TB lesion formation diagnosed by chest radiographic exam. Bronchoalveolar lavage fluid was taken from both lungs. Twenty-four healthy people were chosen as controls. Pyrosequencing was performed on the V3 hypervariable region of 16S rDNA in all bacterial samples and used as a culture-independent method to describe the phylogenetic composition of the microbiota. Through pyrosequencing, 271,764 amplicons were detected in samples and analysed using tools in the Ribosomal Database Project (RDP) and bioinformatics. These analyses revealed significant differences in the microbiota in the lower respiratory tract (LRT) of TB patients compared with healthy controls; in contrast, the microbiota of intra/extra-TB lesions were similar. These results showed that the dominant bacterial genus in the LRT of TB patients was Cupriavidus and not Streptococcus, which resulted in a significant change in the microbiota in TB patients. The abundance of Mycobacteria and Porphyromonas significantly increased inside TB lesions when compared with non-lesion-containing contralateral lungs. From these data, it can be concluded that Cupriavidus plays an important role in TB’s secondary infection and that in addition to Mycobacteria, Porphyromonas may also be a co-factor in lesion formation. The mechanisms underlying this connection warrant further research. PMID:26000957

  1. Lipopolysaccharide found in aseptic loosening of patients with inflammatory arthritis.

    Science.gov (United States)

    Nalepka, Jennifer L; Lee, Michael J; Kraay, Matthew J; Marcus, Randall E; Goldberg, Victor M; Chen, Xin; Greenfield, Edward M

    2006-10-01

    Aseptic loosening of orthopaedic implants occurs in the absence of clinical signs of infection. Nevertheless, bacterial endotoxins derived from subclinical infections, systemic sources, or the implant manufacturing process may contribute to aseptic loosening. Also, the rate of implant infection is greater in patients with inflammatory arthritis than in patients with osteoarthritis. We hypothesized that lipopolysaccharide, the classic endotoxin derived from gram-negative bacteria, is more prevalent in periprosthetic tissue surrounding aseptically loose implants in patients with inflammatory arthritis than in patients with osteoarthritis. To test this, we used a modified Limulus amebocyte assay not affected by beta-glucan-like molecules in mammalian tissues. Lipopolysaccharide rarely was detected in periprosthetic tissue from patients with osteoarthritis and aseptic loosening (one of six patients). In contrast, lipopolysaccharide was detected despite the absence of any clinical signs of infection in peri-prosthetic tissue from all four patients with inflammatory arthritis (rheumatoid arthritis, juvenile rheumatoid arthritis, and systemic lupus erythematosus). Lipopolysaccharide also was detected in two patients with gram-negative infections, who were included as positive control subjects. Endotoxins derived from low-grade or systemic bacteremia may be important contributors to aseptic loosening particularly in patients with autoimmune conditions such as inflammatory arthritis. PMID:16735873

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

    Scientific Electronic Library Online (English)

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

    2010-11-01

    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

  3. CHARACTERIZATION OF AN INTRAVENOUS LIPOPOLYSACCHARIDE INFLAMMATION MODEL IN BROILER CHICKENS

    OpenAIRE

    De Boever, Sandra; Croubels, Siska; Meyer, Evelyne; Sys, Stanislas; Beyaert, Rudi; Ducatelle, Richard; De Backer, Patrick

    2009-01-01

    Abstract Intravenous administration of lipopolysaccharide (LPS) from Escherichia coli O127:B8 at a dose of 1,500,000 units/kg BW evoked a hypothermic response followed by a fever phase in five week old broiler chickens. The hypothermic phase coincided with a severe decrease in blood pressure. We assume that this decrease in blood pressure is, at least partly, responsible for the hypothermic phase of the body temperature curve. LPS administration also caused a decrease in circulatin...

  4. Recruitment of mitochondrial uncoupling protein UCP2 after lipopolysaccharide induction.

    Czech Academy of Sciences Publication Activity Database

    R?ži?ka, Michal; Škobisová, Eva; Dlasková, Andrea; Šantorová, Jitka; Smolková, Katarína; Špa?ek, Tomáš; Žá?ková, Markéta; Modrianský, M.; Ježek, Petr

    2005-01-01

    Ro?. 37, ?. 4 (2005), s. 809-821. ISSN 1357-2725 R&D Projects: GA ?R(CZ) GA301/02/1215; GA ?R(CZ) GP301/01/P084; GA AV ?R(CZ) IAA5011106 Grant ostatní: NIH(US) TW01487 Institutional research plan: CEZ:AV0Z5011922 Keywords : lipopolysaccharide * oxidative stress in liver * mitochondrial uncoupling protein UCP2 Subject RIV: CE - Biochemistry Impact factor: 3.871, year: 2005

  5. Purification and characterization of murine lipopolysaccharide-binding protein.

    OpenAIRE

    Gallay, P.; Carrel, S.; Glauser, M. P.; Barras, C.; Ulevitch, R. J.; Tobias, P. S.; Baumgartner, J. D.; Heumann, D.

    1993-01-01

    The serum protein lipopolysaccharide (LPS)-binding protein (LBP) seems to play an important role in regulating host responses to LPS. Complexes of LPS and LBP form in serum and stimulate monocytes, macrophages, or polymorphonuclear leukocytes after binding to CD14. Previous reports have described the structure and properties of LBP from human and rabbit sera. Since mice are used in some experimental models of endotoxemia or gram-negative bacterial infections, information is needed about the p...

  6. Bacterial lipopolysaccharide induction of leukocyte-derived corticotropin and endorphins.

    OpenAIRE

    Harbour-mcmenamin, D.; Smith, E. M.; Blalock, J. E.

    1985-01-01

    Previous reports have shown that there is an endogenous opioid component associated with pathophysiological responses to endotoxin. It has been shown that these responses are alleviated by naloxone, a specific opiate antagonist. Results of another study have indicated that leukocytes may mediate some of those responses since leukocyte depletion alleviated the effects of lipopolysaccharide. In view of the above reports as well as the finding that leukocytes produce immunoreactive (ir-) endorph...

  7. Lipopolysaccharide Induces Disseminated Endothelial Apoptosis Requiring Ceramide Generation

    OpenAIRE

    Haimovitz-friedman, Adriana; Cordon-cardo, Carlos; Bayoumy, Shariff; Garzotto, Mark; Mcloughlin, Maureen; Gallily, Ruth; Edwards, Carl K.; Schuchman, Edward H.; Fuks, Zvi; Kolesnick, Richard

    1997-01-01

    The endotoxic shock syndrome is characterized by systemic inflammation, multiple organ damage, circulatory collapse and death. Systemic release of tumor necrosis factor (TNF)-? and other cytokines purportedly mediates this process. However, the primary tissue target remains unidentified. The present studies provide evidence that endotoxic shock results from disseminated endothelial apoptosis. Injection of lipopolysaccharide (LPS), and its putative effector TNF-?, into C57BL/6 mice induc...

  8. Lipopolysaccharide induces a fibrotic-like phenotype in endothelial cells

    OpenAIRE

    Echeverri?a, Ce?sar; Montorfano, Ignacio; Sarmiento, Daniela; Becerra, Alvaro; Nun?ez-villena, Felipe; Figueroa, Xavier F.; Cabello-verrugio, Claudio; Elorza, Alvaro A.; Riedel, Claudia; Simon, Felipe

    2013-01-01

    Endothelial dysfunction is crucial in endotoxaemia-derived sepsis syndrome pathogenesis. It is well accepted that lipopolysaccharide (LPS) induces endothelial dysfunction through immune system activation. However, LPS can also directly generate actions in endothelial cells (ECs) in the absence of participation by immune cells. Although interactions between LPS and ECs evoke endothelial death, a significant portion of ECs are resistant to LPS challenge. However, the mechanism that confers endo...

  9. Pleurotus eryngii Ameliorates Lipopolysaccharide-Induced Lung Inflammation in Mice

    OpenAIRE

    Junya Kawai; Tsugunobu Andoh; Kenji Ouchi; Satoshi Inatomi

    2014-01-01

    Pleurotus eryngii (P. eryngii) is consumed as a fresh cultivated mushroom worldwide and demonstrated to have multiple beneficial effects. We investigated the anti-inflammatory effect of P. eryngii in mice with acute lung injury (ALI). Intranasal instillation of lipopolysaccharide (LPS) (10??g/site/mouse) induced marked lung inflammation (increase in the number of inflammatory cells, protein leakage, and production of nitric oxide in bronchoalveolar lavage fluid) as well as histopathologica...

  10. Curcumin Attenuation of Lipopolysaccharide Induced Cardiac Hypertrophy in Rodents

    OpenAIRE

    Rupak Chowdhury; Ramadevi Nimmanapalli; Thomas Graham; Gopal Reddy

    2013-01-01

    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...

  11. 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)

    Gabriella Di Francesco

    2012-12-01

    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.

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

    Science.gov (United States)

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

    2014-11-01

    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 (PLactobacillus reuteri in ileal digesta than pigs fed the other diets. 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

  13. 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. ... 2006 Jan;86(1):9-22. (.png) (.svg) (.html ) (.csml) Show Lipopolysaccharide signaling in endo ... 1):9-22. Pathway - PNG File (.png) SVG File (.svg) HTML ... File (.html ) CSML File (.csml) Open .csml file wit ...

  14. Acute lipopolysaccharide administration impaired imune responsiveness in normal rats

    Directory of Open Access Journals (Sweden)

    Marius Stefan

    2009-06-01

    Full Text Available Involving of endotoxin lipopolysaccharide (LPS on immune response modulation was examined in adult male Wistar rats. Acute LPS injection (25?g/kg, i.p, Sigma increased serum corticosterone, suggesting significant effects on hypothalamic-pituitary-adrenal axis (HPAA activity. In addition, acute LPS administration significantly decreased the number of leukocyte, the number of lymphocyte, total serum protein, antibody titer, body weight and significantly increased albumin-globulin ratio, suggesting that LPS significantly impaired immune responsiveness. Taken together, our data provide further support for modulation of immune response during acute LPS administration.

  15. Lipopolysaccharide induces expression of collagen VI in the rat lung

    OpenAIRE

    Okawa, Sayuri; Unuma, Kana; Yamada, Atsushi; Aki, Toshihiko; Uemura, Koichi

    2014-01-01

    The involvement of the lung during the septic systemic inflammatory response elicited by administration of lipopolysaccharide (LPS) was investigated. Eight-week-old male Sprague–Dawley rats were injected i.p. with 15 mg/kg LPS. After 24 h, the lungs were excised to evaluate the cellular responses to LPS. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF) analysis revealed that type VI collagen (ColVI) was extremely upregulated during sepsis in the rat lun...

  16. 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 ... :186-92. (.png) (.svg) (.html) (.csml) Show Innate recognition ... of lipopolysaccharide by Toll-like receptor 4-MD-2 ... . PubmedID 15051069 Title Innate recognition ... of lipopolysaccharide by Toll-like receptor 4-MD-2 ...

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

    Directory of Open Access Journals (Sweden)

    Bullon Pedro

    2012-10-01

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

  18. Lipopolysaccharide induces amyloid formation of antimicrobial peptide HAL-2.

    Science.gov (United States)

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

    2014-11-01

    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

  19. Pleurotus eryngii Ameliorates Lipopolysaccharide-Induced Lung Inflammation in Mice.

    Science.gov (United States)

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

    2014-01-01

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

  20. Host Species-Specific Damage to Oviduct Mucosa by Neisseria gonorrhoeae Lipopolysaccharide

    OpenAIRE

    Gregg, Clark R.; Johnson, Alan P.; Taylor-Robinson, David; Melly, M. Ann; McGee, Zell A.

    1981-01-01

    The selective toxicity of gonococcal lipopolysaccharide for the mucosa of human fallopian tubes, which is demonstrated in these studies, may be responsible in part for the specificity of naturally occurring gonococcal infections for humans.

  1. 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 ... .csml file with CIOPlayer - ?CIO Player???????? ... Open .csml file with CIO Open .csml file with CIO ... - ?CIO???????? ...

  2. Structural Studies of Lipid A from a Lipopolysaccharide of the Coxiella burnetii isolate RSA 514 (Crazy).

    Czech Academy of Sciences Publication Activity Database

    Vadovi?, P.; Fuleová, A.; Ihnatko, R.; Škultéty, L.; Halada, Petr; Toman, R.

    2009-01-01

    Ro?. 15, ?. 2 (2009), s. 198-199. ISSN 1198-743X Institutional research plan: CEZ:AV0Z50200510 Keywords : lipid * lipopolysaccharide * crazy Subject RIV: EE - Microbiology, Virology Impact factor: 4.014, year: 2009

  3. The effect of metronidazole on the presence of P. gingivalis and T. forsythia at 3 and 12 months after different periodontal treatment strategies evaluated in a randomized, clinical trial

    DEFF Research Database (Denmark)

    Preus, Hans R; Gjermo, Per

    2015-01-01

    Abstract Objective. The benefit of full-mouth disinfection (FDIS) over traditional scaling and root planing (SRP) in the treatment of chronic, destructive periodontitis remains equivocal and it is not known whether the use of adjunctive antibiotics may enhance the effect of FDIS. Therefore, the aim of this study was to evaluate the effect of conventional SRP completed over 21 days or 1-day FDIS, with or without systemically delivered adjunctive metronidazole (MET) on the presence of P. gingivalis and T. forsythia after 3 and 12 months. Materials and methods. One hundred and eighty-four patients with moderate-to-severe periodontitis were randomly allocated to one of four treatment groups; (1) FDIS+MET; (2) FDIS+placebo; (3) SRP+MET; (4) SRP+placebo. Prior to treatment, pooled subgingival samples were obtained from the five deepest pockets. The same sites were sampled again 3 and 12 months after treatment. All samples were analyzed for P. gingivalis and T. forsythia by PCR, whereas A. actinomycetemcomitans and other bacteria were identified by culture techniques. Results. At baseline, 47% of the samples were positive for P. gingivalis, while almost all samples were positive for T. forsythia. The occurrence of P. gingivalis and T. forsythia was significantly reduced at 3 and 12 months after treatment in the FDIS+MET group, but not in the other treatment groups. Conclusion. FDIS+MET had a significant effect in patients with P. gingivalis and T. forsythia, resulting in a significant reduction in number of patients where these micro-organisms could be detected at 3 and 12 months post-therapy.

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

    OpenAIRE

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

    1991-01-01

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

  5. Pathogen-Mediated Proteolysis of the Cell Death Regulator RIPK1 and the Host Defense Modulator RIPK2 in Human Aortic Endothelial Cells

    OpenAIRE

    Madrigal, Andre?s G.; Barth, Kenneth; Papadopoulos, George; Genco, Caroline Attardo

    2012-01-01

    Porphyromonas gingivalis is the primary etiologic agent of periodontal disease that is associated with other human chronic inflammatory diseases, including atherosclerosis. The ability of P. gingivalis to invade and persist within human aortic endothelial cells (HAEC) has been postulated to contribute to a low to moderate chronic state of inflammation, although how this is specifically achieved has not been well defined. In this study, we demonstrate that P. gingivalis infection of HAEC resul...

  6. 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

  7. Toll-Like Receptor 9-Mediated Inflammation Triggers Alveolar Bone Loss in Experimental Murine Periodontitis.

    Science.gov (United States)

    Kim, Paul D; Xia-Juan, Xia; Crump, Katie E; Abe, Toshiharu; Hajishengallis, George; Sahingur, Sinem E

    2015-07-01

    Chronic periodontitis is a local inflammatory disease induced by a dysbiotic microbiota and leading to destruction of the tooth-supporting structures. Microbial nucleic acids are abundantly present in the periodontium, derived through release after phagocytic uptake of microbes and/or from biofilm-associated extracellular DNA. Binding of microbial DNA to its cognate receptors, such as Toll-like receptor 9 (TLR9), can trigger inflammation. In this study, we utilized TLR9 knockout (TLR9(-/-)) mice and wild-type (WT) controls in a murine model of Porphyromonas gingivalis-induced periodontitis and report the first in vivo evidence that TLR9 signaling mediates the induction of periodontal bone loss. P. gingivalis-infected WT mice exhibited significantly increased bone loss compared to that in sham-infected WT mice or P. gingivalis-infected TLR9(-/-) mice, which were resistant to bone loss. Consistent with this, the expression levels of interleukin 6 (IL-6), tumor necrosis factor (TNF), and receptor-activator of nuclear factor kappa B ligand (RANKL) were significantly elevated in the gingival tissues of the infected WT mice but not in infected TLR9(-/-) mice compared to their levels in controls. Ex vivo studies using splenocytes and bone marrow-derived macrophages revealed significantly diminished cytokine production in TLR9(-/-) cells relative to the cytokine production in WT cells in response to P. gingivalis, thereby implicating TLR9 in inflammatory responses to this organism. Intriguingly, compared to the cytokine production in WT cells, TLR9(-/-) cells exhibited significantly decreased proinflammatory cytokine production upon challenge with lipopolysaccharide (LPS) (TLR4 agonist) or Pam3Cys (TLR2 agonist), suggesting possible cross talk between TLR9, TLR4, and TLR2. Collectively, our results provide the first proof-of-concept evidence implicating TLR9-triggered inflammation in periodontal disease pathogenesis, thereby identifying a new potential therapeutic target to control periodontal inflammation. PMID:25964477

  8. Lipopolysaccharide aggravates bisphosphonate-induced osteonecrosis in rats.

    Science.gov (United States)

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

    2015-04-01

    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

  9. Mutual antagonism between antigen- and lipopolysaccharide-induced antibody production.

    Science.gov (United States)

    Anderson, C C; Rahimpour, R; Sinclair, N R

    1993-12-01

    B cells are induced to antibody production by antigens or by mitogens, such as lipopolysaccharide (LPS). We observed a mutually antagonistic relationship between activation through the antigen-receptor (AgR) and LPS-receptor (LPSR) in vitro. Prior exposure of B cells to AgR-ligating antibody prevented antibody forming cell (AFC) production induced by LPS, but not that induced by specific antigen (SRBC, TNP-Ficoll, or TNP-LPS). AFC production induced by antigen could be abrogated by concomitant exposure to LPS; the shutdown of the antigen-driven response was apparent when LPS-induced AFC were prevented by pre-exposure to antibody against the AgR. The ability of signaling through the AgR to inhibit antibody production stimulated by LPS was seen in DBA/2 and BALB/c mouse strains, and not in the New Zealand Black (NZB) strain. The results suggest that mutual antagonism is distinct from other forms of immune hyporesponsiveness, and that defects in antagonism may be a factor in the development of autoimmune disease. PMID:7507885

  10. Purification and characterization of murine lipopolysaccharide-binding protein.

    Science.gov (United States)

    Gallay, P; Carrel, S; Glauser, M P; Barras, C; Ulevitch, R J; Tobias, P S; Baumgartner, J D; Heumann, D

    1993-01-01

    The serum protein lipopolysaccharide (LPS)-binding protein (LBP) seems to play an important role in regulating host responses to LPS. Complexes of LPS and LBP form in serum and stimulate monocytes, macrophages, or polymorphonuclear leukocytes after binding to CD14. Previous reports have described the structure and properties of LBP from human and rabbit sera. Since mice are used in some experimental models of endotoxemia or gram-negative bacterial infections, information is needed about the properties of murine LBP. Murine LBP was purified by ion-exchange chromatography and high-pressure liquid chromatography; its NH2-terminal sequence (TNPGLVTRIT) was very similar to those of human and rabbit LBPs (80 to 90% amino acid identity). Murine LBP resembled LBPs from other species in that it promoted the binding of LPS to monocytes and enhanced the sensitivity of monocytes to LPS at least 100-fold. Mouse LBP, like rabbit and human LBPs, was found to be an acute-phase protein. Further in vivo studies with mice and anti-CD14 or anti-LBP reagents should help determine the role of LBP in response to LPS challenges. Images PMID:7678583

  11. Identification of lipopolysaccharide-binding proteins in porcine milk

    Science.gov (United States)

    Shahriar, Farshid; Gordon, John R.; Simko, Elemir

    2006-01-01

    Septicemia and endotoxemia initiated by bacterial lipopolysaccharide (LPS) are relatively common in suckling and weaned piglets. Maternal milk is a source of both nutrition and immune protection for piglets. Passive transfer of colostral antibodies is necessary for protection of neonatal piglets against diseases, but the concentration of immunoglobulins in milk rapidly declines during the 1st wk of lactation in all mammals. We hypothesized, therefore, that nonimmunoglobulin substances in milk contribute to the innate protection of neonates against septicemia during the suckling period. Using LPS-affinity chromatography for isolation of LPS-binding proteins and liquid chromatography–mass spectrometry for their identification, we identified in porcine milk the following proteins with LPS-binding capacity: lactoferrin, soluble CD14, serum amyloid A, ?-S1 casein, ?-casein, and ?-casein. For lactoferrin, ?-S1 casein, and ?-casein, in vitro pepsin digestion did not inhibit LPS-binding activity, whereas combined digestion with pepsin and pancreatin abolished it. The biologic functions of these LPS-binding proteins and peptides were not determined. PMID:17042375

  12. SHIP prevents lipopolysaccharide from triggering an antiviral response in mice

    Science.gov (United States)

    Sly, Laura M.; Hamilton, Melisa J.; Kuroda, Etsushi; Ho, Victor W.; Antignano, Frann L.; Omeis, Stephanie L.; van Netten-Thomas, Christina J.; Wong, Dana; Brugger, Hayley K.; Williams, Olusegun; Feldman, Morris E.; Houseman, Benjamin T.; Fiedler, Dorothea; Shokat, Kevan M.

    2009-01-01

    Gram-negative bacterial infections, unlike viral infections, do not typically protect against subsequent viral infections. This is puzzling given that lipopolysaccharide (LPS) and double-stranded (ds) RNA both activate the TIR domain–containing adaptor-inducing interferon ? (TRIF) pathway and, thus, are both capable of eliciting an antiviral response by stimulating type I interferon (IFN) production. We demonstrate herein that SH2-containing inositol-5?-phosphatase (SHIP) protein levels are dramatically increased in murine macrophages via the MyD88-dependent pathway, by up-regulating autocrine-acting transforming growth factor-? (TGF?). The increased SHIP then mediates, via inhibition of the phosphatidylinositol-3-kinase (PI3K) pathway, cytosine-phosphate-guanosine (CPG)– and LPS-induced tolerance and cross-tolerance and restrains IFN-? production induced by a subsequent exposure to LPS or dsRNA. Intriguingly, we found, using isoform-specific PI3K inhibitors, that LPS- or cytosine-phosphate-guanosine-induced interleukin-6 (IL-6) is positively regulated by p110?, -?, and -? but negatively regulated by p110?. This may explain some of the controversy concerning the role of PI3K in Toll-like receptor–induced cytokine production. Consistent with our in vitro findings, SHIP?/? mice overproduce IFN-? in response to LPS, and this leads to antiviral hypothermia. Thus, up-regulation of SHIP in response to Gram-negative bacterial infections probably explains the inability of such infections to protect against subsequent viral infections. PMID:19139077

  13. Self-assembly of lipopolysaccharide layers on allantoin crystals.

    Science.gov (United States)

    Vagenende, Vincent; Ching, Tim-Jang; Chua, Rui-Jing; Jiang, Qiu Zhen; Gagnon, Pete

    2014-08-01

    Self-assembly of lipopolysaccharides (LPS) on solid surfaces is important for the study of bacterial membranes, but has not been possible due to technical difficulties and the lack of suitable solid supports. Recently we found that crystals of the natural compound allantoin selectively bind pure LPS with sub-nanomolar affinity. The physicochemical origins of this selectivity and the adsorption mode of LPS on allantoin crystals remain, however, unknown. In this study we present evidence that LPS adsorption on allantoin crystals is initiated through hydrogen-bond attachment of hydrophilic LPS regions. Hydrophobic interactions between alkyl chains of adjacently adsorbed LPS molecules subsequently promote self-assembly of LPS layers. The essential role of hydrogen-bond interactions is corroborated by our finding that allantoin crystals bind to practically any hydrophilic surface chemistry. Binding contributions of hydrophobic interactions between LPS alkyl chains are evidenced by the endothermic nature of the adsorption process and explain why the binding affinity for LPS is several orders of magnitude higher than for proteins (lysozyme, BSA and IgG) and polysaccharides. Self-assembly of LPS layers via hydrogen-bond attachment on allantoin crystals emerges as a novel binding mechanism and could be considered as a practical method for preparing biomimetic membranes on a solid support. PMID:24905674

  14. Lipopolysaccharide induced inflammation in the perivascular space in lungs

    Directory of Open Access Journals (Sweden)

    Pabst Reinhard

    2008-07-01

    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.

  15. Arctigenin attenuates lipopolysaccharide-induced acute lung injury in rats.

    Science.gov (United States)

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

    2015-04-01

    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

  16. A method for generating pulmonary neutrophilia using aerosolized lipopolysaccharide.

    Science.gov (United States)

    Roos, Abraham B; Berg, Tove; Ahlgren, Kerstin M; Grunewald, Johan; Nord, Magnus

    2014-01-01

    Acute lung injury (ALI) is a severe disease characterized by alveolar neutrophilia, with limited treatment options and high mortality. Experimental models of ALI are key in enhancing our understanding of disease pathogenesis. Lipopolysaccharide (LPS) derived from gram positive bacteria induces neutrophilic inflammation in the airways and lung parenchyma of mice. Efficient pulmonary delivery of compounds such as LPS is, however, difficult to achieve. In the approach described here, pulmonary delivery in mice is achieved by challenge to aerosolized Pseudomonas aeruginosa LPS. Dissolved LPS was aerosolized by a nebulizer connected to compressed air. Mice were exposed to a continuous flow of LPS aerosol in a Plexiglas box for 10 min, followed by 2 min conditioning after the aerosol was discontinued. Tracheal intubation and subsequent bronchoalveolar lavage, followed by formalin perfusion was next performed, which allows for characterization of the sterile pulmonary inflammation. Aerosolized LPS generates a pulmonary inflammation characterized by alveolar neutrophilia, detected in bronchoalveolar lavage and by histological assessment. This technique can be set up at a small cost with few appliances, and requires minimal training and expertise. The exposure system can thus be routinely performed at any laboratory, with the potential to enhance our understanding of lung pathology. PMID:25548888

  17. Alterations in Caenorhabditis elegans and Cronobacter sakazakii lipopolysaccharide during interaction.

    Science.gov (United States)

    Sivamaruthi, Bhagavathi Sundaram; Prasanth, Mani Iyer; Balamurugan, Krishnaswamy

    2015-03-01

    Lipopolysaccharide is one of the pathogen-associated molecular patterns of Gram-negative bacteria which are essential for its pathogenicity. Cronobacter sakazakii is an opportunistic, emergent pathogen, which infects and cause mortality in Caenorhabditis elegans. In this study, modifications in host and C. sakazakii LPS during infections were evaluated. The physiological assays revealed that LPS alone is sufficient to affect the host pharyngeal pumping rate, brood size and cause lethality. FTIR spectra of LPS revealed that C. sakazakii modifies its LPS to escape from the recognition of host immune system. These results indicate that LPS plays a key role in C. sakazakii pathogenicity. qPCR studies revealed that LPS modulated the expression of selected host immune (clec-60, clec-87, lys-7, ilys-3, F08G5.6, atf-7, scl-2, cpr-2) and aging-related genes (skn-1, clk-2, bra-2, age-1, bec-1, daf-16, daf-2). Moreover, it was confirmed that p38 MAPK pathway has a major role in host immune response against LPS-mediated challenges. PMID:25416126

  18. 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.

  19. Lipopolysaccharide surface structure does not influence IcsA polarity.

    Science.gov (United States)

    Doyle, Matthew Thomas; Grabowicz, Marcin; May, Kerrie Leanne; Morona, Renato

    2015-04-01

    Shigella species are the causative agents of human bacillary dysentery. These bacteria spread within the lining of the gut via a process termed actin-based motility whereby an actin 'tail' is formed at the bacterial pole. The bacterial outer membrane protein IcsA initiates this process, and crucially is precisely positioned on the bacterial polar surface. Lipopolysaccharide (LPS) O-antigen surface structure has been implicated as an augmenting factor of polarity maintenance due to the apparent dysregulation of IcsA polarity in O-antigen deficient strains. Due to Shigellae having long and short O-antigen chains on their surfaces, it has been proposed that O-antigen chain lengths are asymmetrically distributed to optimize IcsA exposure at the pole and mask exposure laterally. Additionally, it has been proposed that LPS O-antigen restricts IcsA diffusion from the pole by maintaining minimal membrane fluidity. This study utilizes minicells and quantitative microscopy providing data refuting the models of asymmetric masking and membrane diffusion, and supporting a model of symmetric masking of IcsA. We contend that IcsA surface distribution is equivalent between wild-type and O-antigen deficient strains, and that differences in cellular IcsA levels have confounded previous conclusions. PMID:25778729

  20. Genomic and Proteomic Studies on Plesiomonas shigelloides Lipopolysaccharide Core Biosynthesis

    Science.gov (United States)

    Aquilini, Eleonora; Merino, Susana; Regué, Miguel

    2014-01-01

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

  1. Roles of different forms of lipopolysaccharides in Ralstonia solanacearum pathogenesis.

    Science.gov (United States)

    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

    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

  2. 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

  3. Lipopolysaccharide induces expression of collagen VI in the rat lung.

    Science.gov (United States)

    Okawa, Sayuri; Unuma, Kana; Yamada, Atsushi; Aki, Toshihiko; Uemura, Koichi

    2015-01-01

    The involvement of the lung during the septic systemic inflammatory response elicited by administration of lipopolysaccharide (LPS) was investigated. Eight-week-old male Sprague-Dawley rats were injected i.p. with 15 mg/kg LPS. After 24 h, the lungs were excised to evaluate the cellular responses to LPS. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF) analysis revealed that type VI collagen (ColVI) was extremely upregulated during sepsis in the rat lung within the first 24 h of LPS administration. Upregulation of ColVI protein and its mRNA was demonstrated by Western blot analysis, real time PCR, and immunohistochemistry. To the best of our knowledge, this is the first report demonstrating the activation of ColVI in the rat lung at the early stage of systemic inflammation. Activation of ColVI might be involved in sepsis-mediated lung fibrosis at an early stage. PMID:26023260

  4. 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

  5. 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)

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

    2009-12-01

    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.

  6. Removal of lipopolysaccharide from protein solution using nanostructured porous supports bearing lipid membranes

    Science.gov (United States)

    Wakita, Masa-aki

    2013-11-01

    Polymeric lipid membranes of N-octadecylchitosan, which consists of 70 mol% of 2-(octadecylamino)-2-deoxy- d-glucopyranose, 17 mol% of 2-amino-2-deoxy- d-glucopyranose, and 13 mol% of 2-acetamido-2-deoxy- d-glucopyranose, were covalently immobilized to carboxylated porous supports composed of chitosan and used for the adsorption of pyrogenic lipopolysaccharide. When human serum albumin solution, including 5 mg mL-1 of albumin and 5.6 ng mL-1 of lipopolysaccharide, was passed through a column packed with the resulting porous supports bearing lipid membranes assembled in nanoscale, lipopolysaccharide was removed to as low as a detection limit of 0.020 ng mL-1 with a quantitative recovery of protein. On the other hand, in the case of directly N-octadecylated porous supports having cationic and hydrophobic ligands which are not assembled as lipid membranes, lipopolysaccharide could not be removed to the detection limit and protein recovery was lower than the porous supports bearing lipid membranes. The difference above as well as difference from conventional adsorbents suggested that the selectivity was attributable to an interaction between the cationic lipid membranes of N-octadecylchitosan and lipopolysaccharide as well as protein. The porous supports bearing lipid membranes were stable in 0.5 M NaOH and 0.1 M HCl at ambient temperature. Considering the confirmed excellent selectivity and chemical stability, their practical use as separation media in the pharmaceutical manufacturing can be expected.

  7. The heat sensitivity of cytokine-inducing effect of lipopolysaccharide.

    Science.gov (United States)

    Gao, Baochong; Wang, Yun; Tsan, Min-Fu

    2006-08-01

    Heat inactivation by boiling has been widely used as a criterion to determine whether the observed effects of a protein preparation are a result of lipopolysaccharide (LPS) contamination. However, the heat sensitivity of LPS cytokine-inducing activity has not been characterized. In the current study, we demonstrated that the endotoxin activity, i.e., Limulus amebocyte lysate-gelating activity, and the tumor necrosis factor alpha (TNF-alpha)-inducing activity of LPS (Escherichia coli K-12 JM83, K-12 LCD25, and F583) were sensitive to boiling. Heat treatment by boiling for 15 min was sufficient to inactivate approximately 90% of the LPS TNF-alpha-inducing activity. The heat-induced inactivation of LPS activities was not a result of adherence of boiled LPS to the wall of the container, i.e., polypropylene tubes, or aggregation of boiled LPS. In addition, boiled LPS retained its ability to bind polymyxin B. The presence of protein (ovalbumin) in LPS did not affect the heat sensitivity of LPS. Conversely, boiling reduced the size of LPS aggregates as determined by electrophoresis using native polyacrylamide gel. Likewise, the TNF-alpha-inducing activity of diphosphoryl lipid A (DPLA) was also sensitive to boiling. Thin-layer chromatographic analysis of boiled DPLA revealed that the heat-induced inactivation of DPLA TNF-alpha-inducing activity was not a result of its conversion to monophosphoryl lipid A. We conclude that the TNF-alpha-inducing activity of LPS and DPLA is sensitive to boiling and suggest that heat sensitivity as an indicator of whether the observed effects of a protein preparation are a result of LPS contamination should be used with caution. PMID:16720829

  8. Lumican overexpression exacerbates lipopolysaccharide-induced renal injury in mice.

    Science.gov (United States)

    Lu, Xiao-Mei; Ma, Ling; Jin, Yu-Nan; Yu, Yan-Qiu

    2015-09-01

    The present study aimed to investigate the role of lumican in mice with endotoxin-induced acute renal failure (ARF). Lumican transgenic mice and wild?type mice were injected with lipopolysaccharide (LPS; 10 mg/kg) to establish a model of ARF. The mice were sacrificed at 24 h and the blood and renal tissue samples were collected. The value of serum creatinine (SCr) and blood urea nitrogen (BUN) were measured to determine renal function. An ELISA was used to determined the concentrations of renal cytokines, including tumor necrosis factor (TNF)?, interleukin (IL)?6, IL?4 and IL?10. The protein expression levels of Toll-like receptor (TLR4) and nuclear factor (NF)?B in renal tissues were assessed using western blot analysis. Terminal deoxynucleotidyl transferase?mediated dUTP nick end labeling was performed to monitor apoptosis of renal tissue. Light microscopy and electron microscopy were used to observe structural changes in the renal tissues. Following the administration of LPS, the SCr and BUN values of mice in the lumican transgenic group were higher compared with those in the control group. The expression levels of renal TLR4, NF?B, TNF?, IL?6, IL?4 and IL?10 were upregulated in the lumican transgenic mice compared with those in the wild?type control group. Apoptosis was detected predominantly on the renal tubule. There was a significant difference in the optical density of apoptotic bodies between the control mice and the lumican transgenic mice. Light and electron microscopy demonstrated more severe renal tissue injury in the lumican transgenic mice compared with that in the control mice. In conclusion, LPS may cause excessive apoptosis in the renal tubular cells via the TLR4 signal transduction pathway, a decrease in the number of renal tubular cells and ARF. Lumican may be important in mice with LPS-induced ARF. PMID:26081832

  9. Lipopolysaccharide induces a fibrotic-like phenotype in endothelial cells

    Science.gov (United States)

    Echeverría, César; Montorfano, Ignacio; Sarmiento, Daniela; Becerra, Alvaro; Nuñez-Villena, Felipe; Figueroa, Xavier F; Cabello-Verrugio, Claudio; Elorza, Alvaro A; Riedel, Claudia; Simon, Felipe

    2013-01-01

    Endothelial dysfunction is crucial in endotoxaemia-derived sepsis syndrome pathogenesis. It is well accepted that lipopolysaccharide (LPS) induces endothelial dysfunction through immune system activation. However, LPS can also directly generate actions in endothelial cells (ECs) in the absence of participation by immune cells. Although interactions between LPS and ECs evoke endothelial death, a significant portion of ECs are resistant to LPS challenge. However, the mechanism that confers endothelial resistance to LPS is not known. LPS-resistant ECs exhibit a fibroblast-like morphology, suggesting that these ECs enter a fibrotic programme in response to LPS. Thus, our aim was to investigate whether LPS is able to induce endothelial fibrosis in the absence of immune cells and explore the underlying mechanism. Using primary cultures of ECs and culturing intact blood vessels, we demonstrated that LPS is a crucial factor to induce endothelial fibrosis. We demonstrated that LPS was able and sufficient to promote endothelial fibrosis, in the absence of immune cells through an activin receptor–like kinase 5 (ALK5) activity–dependent mechanism. LPS-challenged ECs showed an up-regulation of both fibroblast-specific protein expression and extracellular matrix proteins secretion, as well as a down-regulation of endothelial markers. These results demonstrate that LPS is a crucial factor in inducing endothelial fibrosis in the absence of immune cells through an ALK5-dependent mechanism. It is noteworthy that LPS-induced endothelial fibrosis perpetuates endothelial dysfunction as a maladaptive process rather than a survival mechanism for protection against LPS. These findings are useful in improving current treatment against endotoxaemia-derived sepsis syndrome and other inflammatory diseases. PMID:23635013

  10. Lipopolysaccharide enhances the cytotoxicity of 2-chloroethyl ethyl sulfide

    Directory of Open Access Journals (Sweden)

    Qui Min

    2003-01-01

    Full Text Available Abstract Background The bacterial endotoxin, lipopolysaccharide (LPS, is a well-characterized inflammatory factor found in the cell wall of Gram-negative bacteria. In this investigation, we studied the cytotoxic interaction between 2-chloroethyl ethyl sulfide (CEES or ClCH2CH2SCH2CH3 and LPS using murine RAW264.7 macrophages. CEES is a sulfur vesicating agent and is an analog of 2,2'-dichlorodiethyl sulfide (sulfur mustard. LPS is a ubiquitous natural agent found in the environment. The ability of LPS and other inflammatory agents (such as TNF-alpha and IL-1beta to modulate the toxicity of CEES is likely to be an important factor in the design of effective treatments. Results RAW 264.7 macrophages stimulated with LPS were found to be more susceptible to the cytotoxic effect of CEES than unstimulated macrophages. Very low levels of LPS (20 ng/ml dramatically enhanced the toxicity of CEES at concentrations greater than 400 ?M. The cytotoxic interaction between LPS and CEES reached a maximum 12 hours after exposure. In addition, we found that tumor necrosis factor-alpha (TNF-alpha and interleukin-1-beta (IL-1-beta as well as phorbol myristate acetate (PMA also enhanced the cytotoxic effects of CEES but to a lesser extent than LPS. Conclusion Our in vitro results suggest the possibility that LPS and inflammatory cytokines could enhance the toxicity of sulfur mustard. Since LPS is a ubiquitous agent in the natural environment, its presence is likely to be an important variable influencing the cytotoxicity of sulfur mustard toxicity. We have initiated further experiments to determine the molecular mechanism whereby the inflammatory process influences sulfur mustard cytotoxicity.

  11. Flow cytometric analysis of crayfish haemocytes activated by lipopolysaccharides

    Science.gov (United States)

    Cardenas, W.; Dankert, J.R.; Jenkins, J.A.

    2004-01-01

    Lipopolysaccharides (LPS) from Gram-negative bacteria are strong stimulators of white river crayfish, Procambarus zonangulus, haemocytes in vitro. Following haemocyte treatment with LPS and with LPS from rough mutant R5 (LPS Rc) from Salmonella minnesota, flow cytometric analysis revealed a conspicuous and reproducible decrease in cell size as compared to control haemocytes. These LPS molecules also caused a reduction in haemocyte viability as assessed by flow cytometry with the fluorescent dyes calcein-AM and ethidium homodimer. The onset of cell size reduction was gradual and occurred prior to cell death. Haemocytes treated with LPS from S. minnesota without the Lipid A moiety (detoxified LPS) decreased in size without a reduction of viability. The action of LPS on crayfish haemocytes appeared to be related to the activation of the prophenoloxidase system because phenoloxidase (PO)-specific activity in the supernatants from control and detoxified LPS-treated cells was significantly lower than that from LPS and LPS-Rc treated cells (P < 0.05). Furthermore, addition of trypsin inhibitor to the LPS treatments caused noticeable delays in cell size and viability changes. These patterns of cellular activation by LPS formulations indicated that crayfish haemocytes react differently to the polysaccharide and lipid A moieties of LPS, where lipid A is cytotoxic and the polysaccharide portion is stimulatory. These effects concur with the general pattern of mammalian cell activation by LPS, thereby indicting commone innate immune recognition mechanisms to bacterial antigens between cells from mammals and invertebrates. These definitive molecular approaches used to verify and identify mechanisms of invertbrate haemocyte responses to LPS could be applied with other glycoconjugates, soluble mediators, or xenobiotic compounds.

  12. Lipopolysaccharide induces a fibrotic-like phenotype in endothelial cells.

    Science.gov (United States)

    Echeverría, César; Montorfano, Ignacio; Sarmiento, Daniela; Becerra, Alvaro; Nuñez-Villena, Felipe; Figueroa, Xavier F; Cabello-Verrugio, Claudio; Elorza, Alvaro A; Riedel, Claudia; Simon, Felipe

    2013-06-01

    Endothelial dysfunction is crucial in endotoxaemia-derived sepsis syndrome pathogenesis. It is well accepted that lipopolysaccharide (LPS) induces endothelial dysfunction through immune system activation. However, LPS can also directly generate actions in endothelial cells (ECs) in the absence of participation by immune cells. Although interactions between LPS and ECs evoke endothelial death, a significant portion of ECs are resistant to LPS challenge. However, the mechanism that confers endothelial resistance to LPS is not known. LPS-resistant ECs exhibit a fibroblast-like morphology, suggesting that these ECs enter a fibrotic programme in response to LPS. Thus, our aim was to investigate whether LPS is able to induce endothelial fibrosis in the absence of immune cells and explore the underlying mechanism. Using primary cultures of ECs and culturing intact blood vessels, we demonstrated that LPS is a crucial factor to induce endothelial fibrosis. We demonstrated that LPS was able and sufficient to promote endothelial fibrosis, in the absence of immune cells through an activin receptor-like kinase 5 (ALK5) activity-dependent mechanism. LPS-challenged ECs showed an up-regulation of both fibroblast-specific protein expression and extracellular matrix proteins secretion, as well as a down-regulation of endothelial markers. These results demonstrate that LPS is a crucial factor in inducing endothelial fibrosis in the absence of immune cells through an ALK5-dependent mechanism. It is noteworthy that LPS-induced endothelial fibrosis perpetuates endothelial dysfunction as a maladaptive process rather than a survival mechanism for protection against LPS. These findings are useful in improving current treatment against endotoxaemia-derived sepsis syndrome and other inflammatory diseases. PMID:23635013

  13. Bacterial lipopolysaccharide promotes profibrotic activation of intestinal fibroblasts.

    LENUS (Irish Health Repository)

    Burke, J P

    2012-02-01

    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.

  14. Sickness behaviour after lipopolysaccharide treatment in ghrelin deficient mice.

    Science.gov (United States)

    Szentirmai, Éva; Krueger, James M

    2014-02-01

    Ghrelin is an orexigenic hormone produced mainly by the gastrointestinal system and the brain. Much evidence also indicates a role for ghrelin in sleep and thermoregulation. Further, ghrelin was recently implicated in immune system modulation. Administration of bacterial lipopolysaccharide (LPS) induces fever, anorexia, and increased non-rapid-eye movement sleep (NREMS) and these actions are mediated primarily by proinflammatory cytokines. Ghrelin reduces LPS-induced fever, suppresses circulating levels of proinflammatory cytokines and reduces the severity and mortality of various models of experimental endotoxemia. In the present study, we determined the role of intact ghrelin signaling in LPS-induced sleep, feeding, and thermoregulatory responses in mice. Sleep-wake activity was determined after intraperitoneal, dark onset administration of 0.4, 2 and 10 ?g LPS in preproghrelin knockout (KO) and wild-type (WT) mice. In addition, body temperature, motor activity and changes in 24-h food intake and body weight were measured. LPS induced dose-dependent increases in NREMS, and suppressed rapid-eye movement sleep, electroencephalographic slow-wave activity, motor activity, food intake and body weight in both Ppg KO and WT mice. Body temperature changes showed a biphasic pattern with a decrease during the dark period followed by an increase in the light phase. The effects of the low and middle doses of LPS were indistinguishable between the two genotypes. Administration of 10 ?g LPS, however, induced significantly larger changes in NREMS and wakefulness amounts, body temperature, food intake and body weight in the Ppg KO mice. These findings support a role for ghrelin as an endogenous modulator of inflammatory responses and a central component of arousal and feeding circuits. PMID:24309634

  15. 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)

    H. H. Arafat

    2009-01-01

    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.

  16. Removal of lipopolysaccharide from protein solution using nanostructured porous supports bearing lipid membranes

    OpenAIRE

    Wakita, Masa-aki

    2013-01-01

    Polymeric lipid membranes of N-octadecylchitosan, which consists of 70 mol% of 2-(octadecylamino)-2-deoxy-d-glucopyranose, 17 mol% of 2-amino-2-deoxy-d-glucopyranose, and 13 mol% of 2-acetamido-2-deoxy-d-glucopyranose, were covalently immobilized to carboxylated porous supports composed of chitosan and used for the adsorption of pyrogenic lipopolysaccharide. When human serum albumin solution, including 5 mg mL-1 of albumin and 5.6 ng mL-1 of lipopolysaccharide, was passed through a col...

  17. Lipid lateral organization on giant unilamellar vesicles containing lipopolysaccharides

    DEFF Research Database (Denmark)

    Kubiak, Jakub; Brewer, Jonathan R.

    2011-01-01

    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.

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

    Scientific Electronic Library Online (English)

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

    2011-04-01

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

  19. Arginine supplementation does not alter nitrogen metabolism of beef steers during a lipopolysaccharide challenge

    Science.gov (United States)

    Demand for arginine (Arg) is reported to increase during immune challenges. This study evaluated effects of lipopolysaccharide (LPS) and abomasal Arg infusion on nitrogen (N) metabolism and immune response of 20 ruminally cannulated steers (369 ± 46 kg BW) in a randomized block design. Each block co...

  20. Influence of the lipopolysaccharide structure of Salmonella enterica serovat Enteritidis on interactions with pig neutrophils.

    Czech Academy of Sciences Publication Activity Database

    Matiašovi?, J.; Št?pánová, H.; Volf, J.; Kubala, Lukáš; Ovesná, P.; Rychlík, I.; Faldyna, M.

    2011-01-01

    Ro?. 150, 1-2 (2011), s. 167-172. ISSN 0378-1135 Grant ostatní: GA MZe(CZ) QH81062 Institutional research plan: CEZ:AV0Z50040507; CEZ:AV0Z50040702 Keywords : Salmonella * pig * lipopolysaccharide Subject RIV: BO - Biophysics Impact factor: 3.327, year: 2011

  1. 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...

  2. Effect of prenatal stress on subsequent response to mixing stress and a lipopolysaccharide challenge in pigs

    Science.gov (United States)

    Sows subjected to prenatal stress have been found to produce offspring that alter the manner in which they respond to stress. Our objective was to determine if exposing a sow to stress altered the response of the offspring to lipopolysaccharide (LPS) at 2 mo of age or their response to mixing stres...

  3. Use of Monoclonal Antibodies to Lipopolysaccharide for Antigenic Analysis of Coxiella burnetii

    OpenAIRE

    Hotta, Akitoyo; Kawamura, Midori; To, Ho; Andoh, Masako; Yamaguchi, Tsuyoshi; Fukushi, Hideto; Amano, Ken-ichi; Hirai, Katsuya

    2003-01-01

    Antigenic differences among Coxiella burnetii strains were analyzed. The monoclonal antibodies against the lipopolysaccharide outer core did not react with the strains containing a QpRS plasmid or with plasmidless strains, whereas they reacted with strains containing a QpH1 or QpDV plasmid. C. burnetii isolates could be divided into two groups immunologically.

  4. The lipopolysaccharide coreceptor CD14 is present and functional in seminal plasma and expressed on spermatozoa.

    Czech Academy of Sciences Publication Activity Database

    Harris, C. L.; Vigar, M. A.; Nores, J. E. R.; Ho?ejší, Václav; Labeta, M. O.; Morgan, B. P.

    2001-01-01

    Ro?. 104, ?. 3 (2001), s. 317-323. ISSN 0019-2805 R&D Projects: GA MŠk LN00A026 Keywords : CD14 * spermatozoa * lipopolysaccharide Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.656, year: 2001

  5. Bradykinin inducible receptor is essential to lipopolysaccharide-induced acute lung injury in mice.

    Science.gov (United States)

    Campanholle, Gabriela; Landgraf, Richardt G; Borducchi, Erica; Semedo, Patricia; Wang, Pamela H M; Amano, Mariane T; Russo, Momtchilo; Pacheco-Silva, Alvaro; Jancar, Sonia; Camara, Niels O S

    2010-05-25

    Lipopolysaccharides from gram-negative bacteria are amongst the most common causative agents of acute lung injury, which is characterized by an inflammatory response, with cellular infiltration and the release of mediators/cytokines. There is evidence that bradykinin plays a role in lung inflammation in asthma but in other types of lung inflammation its role is less clear. In the present study we evaluated the role of the bradykinin B1 receptor in acute lung injury caused by lipopolysaccharide inhalation and the mechanisms behind bradykinin actions participating in the inflammatory response. We found that in C57Bl/6 mice, the bradykinin B1 receptor expression was up-regulated 24h after lipopolysaccharide inhalation. At this time, the number of cells and protein concentration were significantly increased in the bronchoalveolar lavage fluid and the mice developed airway hyperreactivity to methacholine. In addition, there was an increased expression of tumor necrosis factor-alpha, interleukin-1 beta and interferon-gamma and chemokines (monocytes chemotactic protein-1 and KC) in the bronchoalveolar lavage fluid and in the lung tissue. We then treated the mice with a bradykinin B1 receptor antagonist, R-954 (Ac-Orn-[Oic2, alpha-MePhe5, D-betaNal7, Ile8]desArg9-bradykinin), 30 min after lipopolysaccharide administration. We observed that this treatment prevented the airway hyperreactivity as well as the increased cellular infiltration and protein content in the bronchoalveolar lavage fluid. Moreover, R-954 inhibited the expression of cytokines/chemokines. These results implicate bradykinin, acting through B1 receptor, in the development of acute lung injury caused by lipopolysaccharide inhalation. PMID:20153312

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

    Directory of Open Access Journals (Sweden)

    Hussain S

    2012-03-01

    Full Text Available Salik Hussain1,*, Faris Al-Nsour1,*, Annette B Rice1, Jamie Marshburn1, Zhaoxia Ji2, Jeffery I Zink2, Brenda Yingling1, Nigel J Walker3, Stavros Garantziotis11Clinical Research Unit, National Institute of Environmental Health Sciences/National Institute of Health, Research Triangle Park, NC, 2UC Center for Environmental Implications of Nanotechnology University of California, Los Angeles, CA, 3Division of National Toxicology Program, National Institute of Environmental Health Sciences/National Institute of Health, Research Triangle Park, NC, USA*Both are principal authorsBackground: Cerium dioxide (CeO2 nanoparticles have potential therapeutic applications and are widely used for industrial purposes. However, the effects of these nanoparticles on primary human cells are largely unknown. The ability of nanoparticles to exacerbate pre-existing inflammatory disorders is not well documented for engineered nanoparticles, and is certainly lacking for CeO2 nanoparticles. We investigated the inflammation-modulating effects of CeO2 nanoparticles at noncytotoxic concentrations in human peripheral blood monocytes.Methods: CD14+ cells were isolated from peripheral blood samples of human volunteers. Cells were exposed to either 0.5 or 1 µg/mL of CeO2 nanoparticles over a period of 24 or 48 hours with or without lipopolysaccharide (10 ng/mL prestimulation. Modulation of the inflammatory response was studied by measuring secreted tumor necrosis factor-alpha, interleukin-1beta, macrophage chemotactic protein-1, interferon-gamma, and interferon gamma-induced protein 10.Results: CeO2 nanoparticle suspensions were thoroughly characterized using dynamic light scattering analysis (194 nm hydrodynamic diameter, zeta potential analysis (-14 mV, and transmission electron microscopy (irregular-shaped particles. Transmission electron microscopy of CD14+ cells exposed to CeO2 nanoparticles revealed that these nanoparticles were efficiently internalized by monocytes and were found either in vesicles or free in the cytoplasm. However, no significant differences in secreted cytokine profiles were observed between CeO2 nanoparticle-treated cells and control cells at noncytotoxic doses. No significant effects of CeO2 nanoparticle exposure subsequent to lipopolysaccharide priming was observed on cytokine secretion. Moreover, no significant difference in lipopolysaccharide-induced cytokine production was observed after exposure to CeO2 nanoparticles followed by lipopolysaccharide exposure.Conclusion: CeO2 nanoparticles at noncytotoxic concentrations neither modulate pre-existing inflammation nor prime for subsequent exposure to lipopolysaccharides in human monocytes from healthy subjects.Keywords: cerium dioxide, nanoparticle, nanomedicine, inflammation, human monocyte, lipopolysaccharides

  7. Helicobacter pylori lipopolysaccharide modification, Lewis antigen expression, and gastric colonization are cholesterol-dependent

    Directory of Open Access Journals (Sweden)

    McGee David J

    2009-12-01

    Full Text Available Abstract Background Helicobacter pylori specifically takes up cholesterol and incorporates it into the bacterial membrane, yet little is currently known about cholesterol's physiological roles. We compared phenotypes and in vivo colonization ability of H. pylori grown in a defined, serum-free growth medium, F12 with 1 mg/ml albumin containing 0 to 50 ?g/ml cholesterol. Results While doubling times were largely unaffected by cholesterol, other overt phenotypic changes were observed. H. pylori strain SS1 grown in defined medium with cholesterol successfully colonized the stomach of gerbils, whereas SS1 grown without cholesterol failed to colonize. H. pylori lipopolysaccharide often displays Lewis X and/or Y antigens. Expression of these antigens measured by whole-cell ELISA was markedly enhanced in response to growth of strain SS1, 26695, or G27 in cholesterol. In addition, electrophoretic analysis of lipopolysaccharide in wild type G27 and in mutants lacking the O-chain revealed structural changes within the oligosaccharide core/lipid A moieties. These responses in Lewis antigen levels and in lipopolysaccharide profiles to cholesterol availability were highly specific, because no changes took place when cholesterol was substituted by ?-sitosterol or bile salts. Disruption of the genes encoding cholesterol ?-glucosyltransferase or lipid A phosphoethanolamine transferase had no effect on Lewis expression, nor on lipopolysaccharide profiles, nor on the cholesterol responsiveness of these properties. Disruption of the lipid A 1-phosphatase gene eliminated the effect of cholesterol on lipopolysaccharide profiles but not its effect on Lewis expression. Conclusions Together these results suggest that cholesterol depletion leads to aberrant forms of LPS that are dependent upon dephosphorylation of lipid A at the 1-position. A tentative model for the observed effects of cholesterol is discussed in which sequential steps of lipopolysaccharide biogenesis and, independently, presentation of Lewis antigen at the cell surface, depend upon membrane composition. These new findings demonstrate that cholesterol availability permits H. pylori to modify its cell envelope in ways that can impact colonization of host tissue in vivo.

  8. Identification and differentiation of Taylorella equigenitalis and Taylorella asinigenitalis by lipopolysaccharide O-antigen serology using monoclonal antibodies

    OpenAIRE

    Brooks, Brian W.; Lutze-wallace, Cheryl L.; Maclean, Leann L.; Vinogradov, Evgeny; Perry, Malcolm B.

    2010-01-01

    Lipopolysaccharides (LPSs) from Taylorella equigenitalis, the causative agent of contagious equine metritis, and T. asinigenitalis were compared by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Lipopolysaccharide profiles of 11 T. equigenitalis strains were similar, but different from the profiles of 3 T. asinigenitalis strains, and the profiles of 2 T. asinigenitalis strains were similar to each other. The serological specificities of the LPSs from these 14 strains w...

  9. Lovastatin Dose-Dependently Potentiates the Pro-inflammatory Activity of Lipopolysaccharide Both In Vitro and In Vivo

    OpenAIRE

    Zanin, Valentina; Marcuzzi, Annalisa; Kleiner, Giulio; Piscianz, Elisa; MONASTA, LORENZO; Zacchigna, Serena; Crovella, Sergio; Zauli, Giorgio

    2013-01-01

    Since contradictory findings have been reported on potential effects of statins in modulating the inflammatory response, we have analysed the biological activity of lovastatin both in vitro using the Raw 264.7 murine macrophagic cell line and in vivo using BALB/c mice. When added to Raw 264.7 cells in combination with lipopolysaccharide, lovastatin significantly potentiated the release of interleukin-1?, interleukin-6 and interleukin-12 with respect to lipopolysaccharide alone and showed an ...

  10. Core Oligosaccharide of Plesiomonas shigelloides PCM 2231 (Serotype O17 Lipopolysaccharide — Structural and Serological Analysis

    Directory of Open Access Journals (Sweden)

    Anna Maciejewska

    2013-02-01

    Full Text Available The herein presented complete structure of the core oligosaccharide of lipopolysaccharide (LPS P. shigelloides Polish Collection of Microorganisms (PCM 2231 (serotype O17 was investigated by 1H, 13C NMR spectroscopy, mass spectrometry, chemical analyses and serological methods. The core oligosaccharide is composed of an undecasaccharide, which represents the second core type identified for P. shigelloides serotype O17 LPS. This structure is similar to that of the core oligosaccharide of P. shigelloides strains 302-73 (serotype O1 and 7-63 (serotype O17 and differs from these only by one sugar residue. Serological screening of 55 strains of P. shigelloides with the use of serum against identified core oligosaccharide conjugated with bovine serum albumin (BSA indicated the presence of similar structures in the LPS core region of 28 O-serotypes. This observation suggests that the core oligosaccharide structure present in strain PCM 2231 could be the most common type among P. shigelloides lipopolysaccharides.

  11. Dok-1 and Dok-2 are negative regulators of lipopolysaccharide-induced signaling

    OpenAIRE

    Shinohara, Hisaaki; Inoue, Akane; Toyama-sorimachi, Noriko; Nagai, Yoshinori; Yasuda, Tomoharu; Suzuki, Hiromi; Horai, Reiko; Iwakura, Yoichiro; Yamamoto, Tadashi; Karasuyama, Hajime; Miyake, Kensuke; Yamanashi, Yuji

    2005-01-01

    Endotoxin, a bacterial lipopolysaccharide (LPS), causes fatal septic shock via Toll-like receptor (TLR)4 on effector cells of innate immunity like macrophages, where it activates nuclear factor ?B (NF-?B) and mitogen-activated protein (MAP) kinases to induce proinflammatory cytokines such as tumor necrosis factor (TNF)-?. Dok-1 and Dok-2 are adaptor proteins that negatively regulate Ras–Erk signaling downstream of protein tyrosine kinases (PTKs). Here, we demonstrate that LPS rapidly ind...

  12. Separation of plasmid DNA from protein and bacterial lipopolysaccharides using displacement chromatography

    OpenAIRE

    Freitag, Ruth; Vogt, Sabine

    1999-01-01

    The preparation of plasmid DNA at large scale constitutes a pressing problem in bioseparation. This paper describes a first investigation of displacement chromatography as a means to separate plasmid DNA (4.7 kb) from E. coli lipopolysaccharides and protein (holo transferrin), respectively. Displacement chromatography has advantages in this regard, since the substance mixture is resolved into rectangular zones of the individual components rather than into peaks. Thus a higher total concentrat...

  13. Low-dose Lipopolysaccharide Selectively Sensitizes Hypoxic-Ischemia White Matter Injury in the Immature Brain

    OpenAIRE

    Wang, Lan-wan; Chang, Ying-chao; Lin, Chang-yi; Hong, Jau-shyong; Huang, Chao-ching

    2010-01-01

    Little is known about the effects of inflammation and hypoxic ischemia (HI), the two important risk factors for white matter (WM) injury in preterm infants, on neuroinflammation and blood-brain barrier (BBB) damage in the WM that displays selective vulnerability in preterm infants. We investigated whether low-dose lipopolysaccharide (LPS) selectively sensitizes HI WM injury in postpartum (P) day 2 pups by selectively increasing neuroinflammation and BBB damage in the WM. P2 pups received LPS ...

  14. Extraction, Purification and Characterization of Lipopolysaccharide from Escherichia coli and Salmonella typhi

    OpenAIRE

    Rezania, Simin; Amirmozaffari, Noor; Tabarraei, Bahman; Jeddi-Tehrani, Mahmood; Zarei, Omid; Alizadeh, Reza; Masjedian, Faramarz; Zarnani, Amir Hassan

    2011-01-01

    Lipopolysaccharide (LPS) is an important structural component of the outer cell membrane complex of gram negative microorganisms. Its causative role in gram negative bacteria-induced diseases and broad applications in different kinds of cell stimulation experiments provided a conceptual basis for studies directed at the isolation, purification, and detailed chemical characterization of LPS. The main problem with LPS purification protocols is the contamination of the end product with nucleic a...

  15. Milk Thistle Extract and Silymarin Inhibit Lipopolysaccharide Induced Lamellar Separation of Hoof Explants in Vitro

    OpenAIRE

    Nicole Reisinger; Simone Schaumberger; Veronika Nagl; Sabine Hessenberger; Gerd Schatzmayr

    2014-01-01

    The pathogenesis of laminitis is not completely identified and the role of endotoxins (lipopolysaccharides, LPS) in this process remains unclear. Phytogenic substances, like milk thistle (MT) and silymarin, are known for their anti-inflammatory and antioxidant properties and might therefore have the potential to counteract endotoxin induced effects on the hoof lamellar tissue. The aim of our study was to investigate the influence of endotoxins on lamellar tissue integrity and to test if MT an...

  16. Effect of Saccharomyces cerevisiae Fermentation Product on Lactation Performance and Lipopolysaccharide Concentration of Dairy Cows

    OpenAIRE

    Zhang, Rui-yang; Yoon, Ilkyu; Zhu, Wei-yun; Mao, Sheng-yong

    2013-01-01

    To evaluate lactation performance and changes in plasma and fecal lipopolysaccharide (LPS) concentrations in response to the supplementation of Saccharomyces cerevisiae fermentation product (SC), two dairy farms were selected. On each farm, 32 cows in early to mid lactation (21 to 140 DIM) were blocked by parity and days in milk (DIM), and randomly assigned to one of the two treatments within block (Control or 56 g SC/cow/d). Effect of SC on lactation performance (daily) and changes in blood ...

  17. Design, synthesis and evaluation of a new fluorescent probe for measuring polymyxin-lipopolysaccharide binding interactions

    OpenAIRE

    Soon, Rachel L.; Velkov, Tony; Chiu, Francis; Thompson, Philip E; Kancharla, Rashmi; Roberts, Kade; Larson, Ian; Nation, Roger L; Li, Jian

    2010-01-01

    Fluorescence assays employing semi-synthetic or commercial dansyl-polymyxin B, have been widely employed to assess the affinity of polycations, including polymyxins, for bacterial cells and lipopolysaccharide (LPS). The five primary ?-amines on diaminobutyric-acid residues of polymyxin B are potentially derivatized with dansyl-cholride. Mass spectrometric analysis of the commercial product revealed a complex mixture of di- or tetra- dansyl-substituted polymyxin B. We synthesized a mono-subst...

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

    OpenAIRE

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

    2007-01-01

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

  19. S-Adenosylmethionine Inhibits Lipopolysaccharide-Induced Gene Expression via Modulation of Histone Methylation

    OpenAIRE

    Ara, Ainhoa Iglesias; Xia, Meng; Ramani, Komal; Mato, Jose? M.; Lu, Shelly C.

    2008-01-01

    We previously showed that S-adenosylmethionine (SAMe) and its metabolite methylthioadenosine (MTA) blocked lipopolysaccharide (LPS)-induced tumor necrosis factor ? (TNF?) expression in RAW (murine macrophage cell line) and Kupffer cells at the transcriptional level without affecting nuclear factor ? B nuclear binding. However, the exact molecular mechanism or mechanisms of the inhibitory effect were unclear. While SAMe is a methyl donor, MTA is an inhibitor of methylation. SAMe can convert...

  20. Cannabidiol (CBD) Enhances Lipopolysaccharide (LPS)-Induced Pulmonary Inflammation in C57BL/6 Mice

    OpenAIRE

    Karmaus, Peer W.F.; Wagner, James G; Harkema, Jack R.; KAMINSKI, Norbert E.; Kaplan, Barbara L. F.

    2012-01-01

    Cannabidiol (CBD) is a plant-derived cannabinoid that has been predominantly characterized as anti-inflammatory. However, it is clear that immune effects of cannabinoids can vary with cannabinoid concentration, or type or magnitude of immune stimulus. The present studies demonstrate that oral administration of CBD enhanced lipopolysaccharide (LPS)-induced pulmonary inflammation in C57BL/6 mice. The enhanced inflammatory cell infiltrate as observed in bronchoalveolar lavage fluid (BALF) was co...

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

    OpenAIRE

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

    2012-01-01

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

  2. Lipopolysaccharide-induced Pulpitis Up-regulates TRPV1 in Trigeminal Ganglia

    OpenAIRE

    Chung, M. -k; Lee, J.; Duraes, G.; Ro, J. Y.

    2011-01-01

    Tooth pain often accompanies pulpitis. Accumulation of lipopolysaccharides (LPS), a product of Gram-negative bacteria, is associated with painful clinical symptoms. However, the mechanisms underlying LPS-induced tooth pain are not clearly understood. TRPV1 is a capsaicin- and heat-gated nociceptive ion channel implicated in thermosensation and hyperalgesia under inflammation or injury. Although TRPV1 is expressed in pulpal afferents, it is not known whether the application of LPS to teeth mod...

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

    OpenAIRE

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

    1999-01-01

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

  4. Designed ?-Boomerang Antiendotoxic and Antimicrobial Peptides: STRUCTURES AND ACTIVITIES IN LIPOPOLYSACCHARIDE*?

    OpenAIRE

    Bhunia, Anirban; Mohanram, Harini; Domadia, Prerna N.; Torres, Jaume; Bhattacharjya, Surajit

    2009-01-01

    Lipopolysaccharide (LPS), an integral part of the outer membrane of Gram-negative bacteria, is involved in a variety of biological processes including inflammation, septic shock, and resistance to host-defense molecules. LPS also provides an environment for folding of outer membrane proteins. In this work, we describe the structure-activity correlation of a series of 12-residue peptides in LPS. NMR structures of the peptides derived in complex with LPS reveal boomerang-like ?-strand conforma...

  5. Detection of antibodies to Shigella lipopolysaccharide in urine after natural Shigella infection or vaccination.

    OpenAIRE

    Cohen, D.; Orr, N.; Robin, G; Slepon, R; Ashkenazi, S; Ashkenazi, I; Shemer, J.

    1996-01-01

    The purpose of the present study was to explore the possibility of detecting antibodies to Shigella sonnei lipopolysaccharide (LPS) in urine after infection or vaccination. Urinary immunoglobulin A (IgA) and IgG antibodies and specific IgA secretory protein against S. sonnei LPS were measured by enzyme-linked immunosorbent assay (ELISA), after adjustment for urine concentration. A significant antibody level was defined as one above a cutoff value calculated from the geometric mean + 2 standar...

  6. Role of Phosphoglucomutase of Bordetella bronchiseptica in Lipopolysaccharide Biosynthesis and Virulence

    OpenAIRE

    West, Nicholas P.; Jungnitz, Heidrun; Fitter, John T.; Mcarthur, Jason D.; Guzma?n, Carlos A.; Walker, Mark J.

    2000-01-01

    The phosphoglucomutase (PGM)-encoding gene of Bordetella bronchiseptica is required for lipopolysaccharide (LPS) biosynthesis. An insertion mutant of the wild-type B. bronchiseptica strain BB7865 which disrupted LPS biosynthesis was created and characterized (BB7865pgm). Genetic analysis of the mutated gene showed it shares high identity with PGM genes of various bacterial species and forms part of an operon which also encompasses the gene encoding phosphoglucose isomerase. Functional assays ...

  7. Lipopolysaccharide, Tumor Necrosis Factor Alpha, or Interleukin-1? Triggers Reactivation of Latent Cytomegalovirus in Immunocompetent Mice

    OpenAIRE

    Cook, Charles H.; Trgovcich, Joanne; Zimmerman, Peter D.; Zhang, Yingxue; Sedmak, Daniel D.

    2006-01-01

    We have previously shown that cytomegalovirus (CMV) can reactivate in lungs of nonimmunosuppressed patients during critical illness. Our recent work has shown that polymicrobial bacterial sepsis can trigger reactivation of latent murine CMV (MCMV). We hypothesize that MCMV reactivation following bacterial sepsis may be caused by inflammatory mediators. To test this hypothesis, BALB/c mice latently infected with Smith strain MCMV received sublethal intraperitoneal doses of lipopolysaccharide (...

  8. Ginsenoside Rg2 Inhibits Lipopolysaccharide-Induced Adhesion Molecule Expression in Human Umbilical Vein Endothelial Cell

    OpenAIRE

    Cho, Young-Suk; Kim, Chan Hyung; Ha, Tae-Sun; Lee, Sang Jin; Ahn, Hee Yul

    2013-01-01

    Vascular cell adhesion molecule 1 (VCAM-1), intercellular adhesion molecule 1 (ICAM-1), P- and E-selectin play a pivotal role for initiation of atherosclerosis. Ginsenoside, a class of steroid glycosides, is abundant in Panax ginseng root, which has been used for prevention of illness in Korea. In this study, we investigated the mechanism(s) by which ginsenoside Rg2 may inhibit VCAM-1 and ICAM-1 expressions stimulated with lipopolysaccharide (LPS) in human umbilical vein endothelial cell (HUV...

  9. Antigen and Lipopolysaccharide Play Synergistic Roles in the Effector Phase of Airway Inflammation in Mice

    OpenAIRE

    Jung, Yong Woo; Schoeb, Trenton R.; Weaver, Casey T.; Chaplin, David D.

    2006-01-01

    T helper (Th) cells play major roles in orchestrating asthmatic airway inflammation, but the molecular mechanisms controlling Th-cell recruitment to the airways remain incompletely defined. Innate immunity contributes importantly to the recruitment of effector T cells into sites of inflammation. To understand better the role of innate immune signals in the development of airway inflammation, we used a murine model in which lipopolysaccharide (LPS) contaminating the antigen is thought to trigg...

  10. Utilization of fructose and ribose in lipopolysaccharide synthesis by Veillonella parvula.

    OpenAIRE

    Tortorello, M. L.; Delwiche, E. A.

    1983-01-01

    Veillonella parvula, which cannot ferment or incorporate most sugars, incorporated radioactivity from [14C]ribose and [14C]fructose into cellular lipopolysaccharide (LPS) in the presence of lactate as an energy source. It was shown that virtually all of the fructose carbon which was assimilated into LPS material appeared in hydrophilic LPS components, and almost none was assimilated into fatty acid LPS components. The assimilation of lactate carbon into LPS in the presence of fructose was shi...

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

    OpenAIRE

    Sledjeski, Darren D.; Weiner, Ronald M.

    1991-01-01

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

  12. Lack of effect of TNF antibodies on the cardiovascular sequelae of lipopolysaccharide infusion in conscious rats.

    OpenAIRE

    J. Waller; Gardiner, S M; Jose, J; Bennett, T

    1995-01-01

    1. The aims of the present study were to determine the profile of tumour necrosis factor (TNF) release, and the effect of monoclonal antibodies to TNF, on the changes in regional haemodynamics and the responses to vasodilator and vasoconstrictor challenges, during a continuous 24 h low dose infusion of lipopolysaccharide (LPS) in conscious rats. 2. Male Long Evans rats were chronically instrumented for measurement of regional haemodynamics (renal, superior mesenteric and hindquarters) and wer...

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

    OpenAIRE

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

    2013-01-01

    Previous studies have confirmed that oxidized adenosine triphosphate, a P2X7 receptor antagonist, attenuates lipopolysaccharide-mediated microglial activation and inflammatory expression following neuronal damage in rat brain. NaCl and temperature may affect the potency of oxidized adenosine triphosphate. Brilliant blue G is a derivative of a widely used food additive and has little toxicity. This study explored the effects of brilliant blue G, a selective P2X7 receptor antagonist, on microgl...

  14. Lipopolysaccharide O1 Antigen Contributes to the Virulence in Klebsiella pneumoniae Causing Pyogenic Liver Abscess

    OpenAIRE

    Hsieh, Pei-fang; Lin, Tzu-Lung; Yang, Feng-ling; Wu, Meng-Chuan; Pan, Yi-Jiun; Wu, Shih-Hsiung; Wang, Jin-Town

    2012-01-01

    Klebsiella pneumoniae is the common cause of a global emerging infectious disease, community-acquired pyogenic liver abscess (PLA). Capsular polysaccharide (CPS) and lipopolysaccharide (LPS) are critical for this microorganism's ability to spread through the blood and to cause sepsis. While CPS type K1 is an important virulence factor in K. pneumoniae causing PLA, the role of LPS in PLA is not clear. Here, we characterize the role of LPS O antigen in the pathogenesis of K. pneumoniae causing ...

  15. Estrogen-Induced Nongenomic Calcium Signaling Inhibits Lipopolysaccharide-Stimulated Tumor Necrosis Factor ? Production in Macrophages

    OpenAIRE

    Liu, Limin; Zhao, Ying; Xie, Keming; Sun, Xiaodong; Gao, Yuzhen; Wang, Zufeng

    2013-01-01

    Estrogen is traditionally thought to exert genomic actions through members of the nuclear receptor family. Here, we investigated the rapid nongenomic effects of 17?-estradiol (E2) on tumor necrosis factor ? (TNF-?) production following lipopolysaccharide (LPS) stimulation in mouse bone marrow-derived macrophages (BMMs). We found that LPS induced TNF-? production in BMMs via phosphorylation of p38 mitogen-activated protein kinase (MAPK). E2 itself did not affect the MAPK pathway, although ...

  16. Dried Ginger (Zingiber officinalis) Inhibits Inflammation in a Lipopolysaccharide-Induced Mouse Model

    OpenAIRE

    Woong Mo Yang; Mi Hye Kim; Sung-Hoon Kim; Jongki Hong; You Yeon Choi

    2013-01-01

    Objectives. Ginger rhizomes have a long history of human use, especially with regards to their anti-inflammatory properties. However, the mechanisms by which ginger acts on lipopolysaccharide-(LPS-)induced inflammation have not yet been identified. We investigated the anti-inflammatory effects of dried Zingiber officinalis (DZO) on LPS-induced hepatic injury. Methods. ICR mice were given a DZO water extract (100, 1000?mg/kg) orally for three consecutive days. On the third day, they were admin...

  17. Lipopolysaccharide-induced blood brain barrier permeability is enhanced by alpha-synuclein expression

    OpenAIRE

    Jangula, Adam; Murphy, Eric J.

    2013-01-01

    Because ?-synuclein (Snca) is involved in neuroinflammatory response, we determined if its expression altered blood-brain barrier (BBB) permeability. To induce increased BBB permeability, Snca gene-ablated (KO) and wild-type (WT) mice were injected (i.p.) with lipopolysaccharide (LPS). To assess changes in BBB permeability, Evans blue was injected (i.p.) and extravasation into the brain assessed using fluorescence spectroscopy. WT mice had a significant increase in BBB permeability at 1, 3, a...

  18. Membrane potential modulates release of tumor necrosis factor in lipopolysaccharide-stimulated mouse macrophages.

    OpenAIRE

    Haslberger, A; Romanin, C.; Koerber, R

    1992-01-01

    Lipopolysaccharide (LPS)-mediated synthesis of macrophage gene products such as tumor necrosis factor (TNF) is controlled by different signaling pathways. We investigated intracellular free Ca2+ (Ca2+ic) and the membrane potential as early cellular responses to LPS and their role in the synthesis and release of TNF. In peritoneal macrophages and in the RAW 269 mouse macrophage cell line, LPS and its biologically active moiety lipid A stimulated TNF synthesis but exerted no significant effects...

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

    OpenAIRE

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

    1990-01-01

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

  20. Selective induction of metabolic activation programs in peritoneal macrophages by lipopolysaccharide substructures.

    OpenAIRE

    Lehmann, V; Benninghoff, B; Dröge, W

    1991-01-01

    The structural elements of Salmonella typhimurium lipopolysaccharides (LPS) that are able to stimulate peritoneal macrophages to produce increased amounts of prostaglandin E2, ornithine, and citrulline, agents known to modulate immune responses, are described. Two different incomplete lipid A structures which lack the carbohydrate portion, the nonhydroxylated fatty acids lauric acid and myristic acid (lipid A precursor IB), and additional palmitic acid (lipid A precursor IA) stimulated increa...

  1. Atrophic rhinitis in swine: correlation of Pasteurella multocida pathogenicity with membrane protein and lipopolysaccharide patterns.

    OpenAIRE

    Lugtenberg, B.; Boxtel, R.; Jong, M.

    1984-01-01

    Cell envelope proteins and lipopolysaccharides (LPS) of Pasteurella multocida strains associated with atrophic rhinitis in swine were compared by using sodium dodecyl sulfate gel electrophoresis. Among 34 strains, three different types of cell envelope protein patterns, named I (16 strains), II (3 strains), and III (15 strains), could be distinguished. These differences were based on the electrophoretic mobility of the major protein, designated as protein H. Comparison of cell envelope protei...

  2. Separation and characterization of lipopolysaccharide isomers using capillary electrophoresis-electrospray tandem mass spectrometry

    OpenAIRE

    Li, J; Cox, A; Martin, A; Hood, D; Moxon, RE; Thibault, P.; Richards, JC

    2002-01-01

    The electrophoretic separation of trace level lipopolysaccharide (LPS) isoforms and their molecular structure were analyzed using tandem mass spectrometry. A Crystal Model 310 CE instrument was coupled to an API 3000 mass spectrometer via a microlonspray interface. LPS from monoclonal antibody B 5 positive phase variant of N.meningitidis clinical isolates, BZ157 was obtained. Structural informations were obtained using second generation product ions of fragments formed by in-source CID under ...

  3. Comparative virulence of intra- and interstrain lipopolysaccharide variants of Coxiella burnetii in the guinea pig model.

    OpenAIRE

    Moos, A.; Hackstadt, T.

    1987-01-01

    We compared the relative infectivity and virulence of lipopolysaccharide (LPS) variants of the Nine Mile strain of Coxiella burnetii with those of the Priscilla strain, a representative of endocarditis-type strains. In agreement with results of previous studies, Nine Mile phase I (9mi/I) organisms were highly infectious, eliciting seroconversion and fever with inocula containing as few as four organisms. Viable 9mi/I was recovered from the spleens of infected animals 30 days postinfection. Ni...

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2012-04-20

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

  5. Lipopolysaccharide based oral nanocarriers for the improvement of bioavailability and anticancer efficacy of curcumin.

    Science.gov (United States)

    Chaurasia, Sundeep; Patel, Ravi R; Chaubey, Pramila; Kumar, Nagendra; Khan, Gayasuddin; Mishra, Brahmeshwar

    2015-10-01

    Soluthin MD(®), a unique phosphatidylcholine-maltodextrin based hydrophilic lipopolysaccharide, which exhibits superior biocompatibility and bioavailability enhancer properties for poorly water soluble drug(s). Curcumin (CUR) is a potential natural anticancer drug with low bioavailability due to poor aqueous solubility. The study aims at formulation and optimization of CUR loaded lipopolysaccharide nanocarriers (C-LPNCs) to enhance oral bioavailability and anticancer efficacy in colon-26 tumor-bearing mice in vitro and in vivo. The Optimized C-LPNCs demonstrated favorable mean particle size (108±3.4nm) and percent entrapment efficiency (65.29±1.0%). Pharmacokinetic parameters revealed ?130-fold increase in oral bioavailability and cytotoxicity studies demonstrated ?23-fold reduction in 50% cell growth inhibition when treated with optimized C-LPNCs as compared to pure CUR. In vivo anticancer study performed with optimized C-LPNCs showed significant increase in efficacy compared with pure CUR. Thus, lipopolysaccharide nanocarriers show potential delivery strategy to improve oral bioavailability and anticancer efficacy of CUR in the treatment of colorectal cancer. PMID:26076595

  6. Genetic derepression of a developmentally regulated lipopolysaccharide antigen from Rhizobium leguminosarum 3841.

    Science.gov (United States)

    Wood, E A; Butcher, G W; Brewin, N J; Kannenberg, E L

    1989-09-01

    Monoclonal antibody AFRC MAC 203 recognizes a developmentally regulated lipopolysaccharide antigen in Rhizobium leguminosarum bv. viciae 3841. Transposon-induced mutants that constitutively expressed MAC 203 antigen were isolated. These strains were morphologically normal, showed no gross abnormalities in lipopolysaccharide size distribution on sodium dodecyl sulfate-polyacrylamide gels, and induced normal nitrogen-fixing nodules. However, the mutants lacked lipopolysaccharide epitopes recognized by another rat monoclonal antibody, AFRC MAC 281, suggesting that the corresponding epitopes may be interconverted or share a common precursor. In conjugational crosses, the transposon insertion associated with both the loss of MAC 281 antigen and the constitutive expression of MAC 203 antigen showed linkage to the chromosomal rif allele. A derivative of strain 3841 with a deletion spanning the nod-fix region of the symbiotic plasmid showed no altered expression pattern for MAC 203 antigen, suggesting that the relevant genetic determinants map to genomic sites that are not associated with nifA or any known genes on the symbiotic plasmid. PMID:2768182

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

    International Nuclear Information System (INIS)

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

  8. Placental-mediated increased cytokine response to lipopolysaccharides: a potential mechanism for enhanced inflammation susceptibility of the preterm fetus

    Directory of Open Access Journals (Sweden)

    Ross MG

    2012-07-01

    Full Text Available Julie L Boles,1 Michael G Ross,1 Ron Beloosesky,2 Mina Desai,1 Louiza Belkacemi11Department of Obstetrics and Gynecology, Harbor-UCLA Medical Center, Los Angeles Biomedical Research Institute at Harbor-UCLA, David Geffen School of Medicine at UCLA, University of California, Los Angeles, Torrance, CA, USA; 2Department of Obstetrics and Gynecology, Rambam Medical Center, Haifa, IsraelBackground: Cerebral palsy is a nonprogressive motor impairment syndrome that has no effective cure. The etiology of most cases of cerebral palsy remains unknown; however, recent epidemiologic data have demonstrated an association between fetal neurologic injury and infection/inflammation. Maternal infection/inflammation may be associated with the induction of placental cytokines that could result in increased fetal proinflammatory cytokine exposure, and development of neonatal neurologic injury. Therefore, we sought to explore the mechanism by which maternal infection may produce a placental inflammatory response. We specifically examined rat placental cytokine production and activation of the Toll-like receptor 4 (TLR4 pathway in response to lipopolysaccharide exposure at preterm and near-term gestational ages.Methods: Preterm (e16 or near-term (e20 placental explants from pregnant rats were treated with 0, 1, or 10 µg/mL lipopolysaccharide. Explant integrity was assessed by lactate dehydrogenase assay. Interleukin-6 and tumor necrosis alpha levels were determined using enzyme-linked immunosorbent assay kits. TLR4 and phosphorylated nuclear factor kappa light chain enhancer of activated B cells (NF?B protein expression levels were determined by Western blot analysis.Results: At both e16 and e20, lactate dehydrogenase levels were unchanged by treatment with lipopolysaccharide. After exposure to lipopolysaccharide, the release of interleukin-6 and tumor necrosis alpha from e16 placental explants increased by 4-fold and 8–9-fold, respectively (P < 0.05 versus vehicle. Conversely, interleukin-6 release from e20 explants was not significantly different compared with vehicle, and tumor necrosis alpha release was only 2-fold higher (P < 0.05 versus vehicle following exposure to lipopolysaccharide. Phosphorylated NF?B protein expression was significantly increased in the nuclear fraction from placental explants exposed to lipopolysaccharide at both e16 and e20, although TLR4 protein expression was unaffected.Conclusion: Lipopolysaccharide induces higher interleukin-6 and tumor necrosis alpha expression at e16 versus e20, suggesting that preterm placentas may have a greater placental cytokine response to lipopolysaccharide infection. Furthermore, increased phosphorylated NF?B indicates that placental cytokine induction may occur by activation of the TLR4 pathway.Keywords: cytokines, lipopolysaccharide, NF?B, placenta, rat pregnancy

  9. Comparison of quantitative and qualitative antibody-producing cell responses to lipopolysaccharide in cell walls of the bacterial form and in membranes of the protoplast L-form of Proteus mirabilis.

    OpenAIRE

    Karch, H.; Nixdorff, K.

    1980-01-01

    Membranes of the stable protoplast L-form of Proteus mirabilis strain VI were highly immunogenic carriers of lipopolysaccharide when compared with the immune responses to lipopolysaccharide contained in cell walls of the bacterial form of this organism.

  10. Serological Evaluation of Brucella abortus S99 Lipopolysaccharide Extracted by an Optimized Method

    Directory of Open Access Journals (Sweden)

    Ali S. Salmani

    2009-01-01

    Full Text Available Problem statement: Brucellosis is a globally found infectious disease and there is no licensed vaccine against human brucellosis. The present study carried-out to evaluate the potency of our modified extracted lipopolysaccharide (LPS of B. abortus to elicit specific anti-Brucella antibodies in animal model (Rabbit as a part of a candidate vaccine for brucellosis. Lipopolysaccharide is one of the main virulence factors and the most immunogenic structure of smooth strains of Brucella. Approach: Lipopolysaccharide of B. abortus S99 (S-LPS initially extracted through an optimized method as described previously. After biochemical and pyrogenicity evaluations of the extracted S-LPS humoral immune response against the extracted LPS analyzed in animal model through serological assays such as Rose Bengal assay, Rapid agglutination (Rapid Wright test and Standard agglutination test (SAT or Wright test to demonstrate the specific elicited antibodies against the injected LPS. In addition, the interaction of LPS and anti-LPS antibodies was demonstrated by Agarose Gel Immunodiffusion (AGID assay. Results: Higher doses of B. abortus S99 LPS caused less or equal body temperature increase in comparison to E. coli LPS doses. Sera of immunized animals had been reported positive by RBT because of B. abortus LPS immunogenicity which we extracted through our optimized method. The highest titer of anti-Brucella antibodies detected two weeks after the third immunization (assayed by rapid slide agglutination and standard agglutination tests. Anti-Brucella antibodies of immunized animals reacted more specifically with the LPS of B. abortus in comparison with E. coli LPS and precipitation lines between B. abortus LPS and immune sera appeared after 30 min while detected after three hours for E. coli LPS. Conclusions/Recommendations: The properties of B. abortus S99 LPS concluded from the present study results, suggest the possible use of this component as a carrier or a part of a sub-unit or conjugated vaccine for human brucellosis.

  11. Effect of passive immunization by anti-gingipain IgY on periodontal health of dogs

    OpenAIRE

    Van Sa Nguyen; Yoshikatsu Kodama; Kouji Umeda; El-Sayed M. Ibrahim; Rahman A.K.M. Shofiqur; Rie Isoda

    2011-01-01

    Anti-gingipain IgY (IgY-GP), known as hyperimmune ?-livetin from egg yolk, inhibits the enzyme activity, growth and adherence of Porphyromonas gingivalis to gingival epithelial cells. Our objective was to evaluate the efficacy of IgY-GP on periodontal health of dogs. IgY-GP was prepared from the egg yolk of hens immunized with the gingipain from Porphyromonas gingivalis ATCC 33277. Two in vivo trial models were conducted on 15 adult dogs with periodontitis by giving IgY-GP-supplemented dog fe...

  12. Effect of gamma irradiation on chemical and biological properties of lipopolysaccharide from Salmonella typhimurium

    International Nuclear Information System (INIS)

    Lipopolysaccharide (LPS) from S. typhimurium on exposure to ?-radiation resulted in decrease in toxicity and was less mitogenic. Silver stained profiles of irradiated LPS on polyacrylamide gels revealed complete loss of its heteropolysaccharides which was confirmed further by analysing lipid A and LPS from Salmonella minnesota Re mutants on SDS-PAGE. Glucosamine and 2-keto 3-deoxy-octonate (Kdo) contents were significantly decreased on treatment. Lipid A obtained by removal of heteropolysaccharides from LPS was less toxic on exposure to gamma radiations. (author)

  13. Characterization of the lipopolysaccharide from a Rhizobium phaseoli mutant that is defective in infection thread development.

    OpenAIRE

    Carlson, R. W.; Kalembasa, S.; Turowski, D.; Pachori, P.; Noel, K. D.

    1987-01-01

    The lipopolysaccharide (LPS) from a Rhizobium phaseoli mutant, CE109, was isolated and compared with that of its wild-type parent, CE3. A previous report has shown that the mutant is defective in infection thread development, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis shows that it has an altered LPS (K. D. Noel, K. A. VandenBosch, and B. Kulpaca, J. Bacteriol. 168:1392-1462, 1986). Mild acid hydrolysis of the CE3 LPS released a polysaccharide and an oligosaccharide, PS1 an...

  14. Requirement for lipopolysaccharide-responsive macrophages in galactosamine-induced sensitization to endotoxin.

    OpenAIRE

    Freudenberg, M A; Keppler, D.; Galanos, C.

    1986-01-01

    Injection of D-galactosamine sensitizes mice many thousand-fold to the lethal action of endotoxin (lipopolysaccharide [LPS]). Comparable sensitization was practically absent in LPS-resistant C3H/HeJ mice, which after D-galactosamine treatment were about 500,000 times less sensitive to LPS lethality than histocompatible LPS-sensitive C3H/HeN mice. D-Galactosamine induces changes in the hepatocytes of treated animals, such as depletion of UTP and alterations in the pattern of UDP sugars. These ...

  15. Identification of living Legionella pneumophila using species-specific metabolic lipopolysaccharide labeling.

    Science.gov (United States)

    Mas Pons, Jordi; Dumont, Audrey; Sautejeau, Grégory; Fugier, Emilie; Baron, Aurélie; Dukan, Sam; Vauzeilles, Boris

    2014-01-27

    Legionella pneumophila is a pathogenic bacterium involved in regular outbreaks characterized by a relatively high fatality rate and an important societal impact. Frequent monitoring of the presence of this bacterium in environmental water samples is necessary to prevent these epidemic events, but the traditional culture-based detection and identification method requires up to 10?days. Reported herein is a method allowing identification of Legionella pneumophila by metabolic lipopolysaccharide labeling which targets, for the first time, a precursor to monosaccharides that are specifically present within the O-antigen of the bacterium. This new approach allows easy detection of living Legionella pneumophila, while other Legionella species are not labeled. PMID:24446310

  16. Susceptibility of porin- and lipopolysaccharide-deficient strains of Escherichia coli to some antiseptics and disinfectants.

    Science.gov (United States)

    Russell, A D; Furr, J R

    1986-07-01

    The sensitivities of some outer membrane protein (Omp) and/or lipopolysaccharide (LPS)-defective mutants of Escherichia coli to chlorhexidine and some quaternary ammonium compounds (QACs) were examined. Wild-type strains, with no defect in Omp or LPS were the most resistant to QACs, whereas LPS-deficient strains were the most sensitive. As expected, QAC resistance could not be related to the presence or absence of any specific porin(s). Chlorhexidine was highly active against all strains, inhibitory concentrations lying within the narrow range of 1 and 2 mg l-1. PMID:2875101

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

    Scientific Electronic Library Online (English)

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

    2014-03-01

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

  18. Production and characterization of monoclonal antibodies against the O-5 antigen of Salmonella typhimurium lipopolysaccharide.

    OpenAIRE

    Jaradat, Z. W.; Zawistowski, J.

    1996-01-01

    Three murine monoclonal antibodies (MAbs) were produced by fusion of P3X63-Ag8.653 myeloma cells and splenocytes of a mouse immunized with heat-attenuated (20 min, 80 degrees C) Salmonella typhimurium cells. MAbs 5A5 and 5B2 were of the immunoglobulin M (IgM) class, while MAb 4A8 was IgG2a. All possessed the kappa light chains. The MAbs were specific to the lipopolysaccharide (LPS) O-5 antigen of Salmonella B serogroup, as determined by electrophoresis followed by immunoblotting. All MAbs rec...

  19. A murine monoclonal antibody specific for the outer core oligosaccharide of Salmonella lipopolysaccharide.

    OpenAIRE

    Tsang, R S; K. H. Chan; Chau, P. Y.; Wan, K C; Ng, M H; Schlecht, S

    1987-01-01

    Immunoglobulin G3 murine monoclonal antibody T6 specific for the lipopolysaccharide of Salmonella O serogroups A to E was established. By using R mutants of Salmonella spp., Escherichia coli, and Shigella spp., the major reactive epitope with T6 was tentatively identified as the terminal disaccharide, N-acetylglucosamine 1.2----alpha glucose, of the core oligosaccharide. T6 was reactive with 10 clinical isolates of each of the Salmonella O serogroups A to E but not with 58 isolates of other g...

  20. Antioxidant properties of lutein contribute to the protection against lipopolysaccharide-induced uveitis in mice

    OpenAIRE

    Yao Xin-Sheng; Yao Nan; Lan Fang; Tsoi Bun; He Rong-Rong; Kurihara Hiroshi

    2011-01-01

    Abstract Background Lutein is an important eye-protective nutrient. This study investigates the protective effects and mechanisms of lutein on lipopolysaccharides (LPS)-induced uveitis in mice. Methods Lutein, suspended in drinking water at a final concentration of 12.5 and 25 mg/mL, was administered to mice at 0.1 mL/10 g body weight for five consecutive days. Control and model group received drinking water only. Uveitis was induced by injecting LPS (100 mg per mouse) into the footpad in the...

  1. Lipopolysaccharides of Brucella abortus and Brucella melitensis Induce Nitric Oxide Synthesis in Rat Peritoneal Macrophages

    OpenAIRE

    Lo?pez-urrutia, Luis; Alonso, Andre?s; Nieto, Maria Luisa; Bayo?n, Yolanda; Ordun?a, Antonio; Sa?nchez Crespo, Mariano

    2000-01-01

    Smooth lipopolysaccharide (S-LPS) and lipid A of Brucella abortus and Brucella melitensis induced the production of nitric oxide (NO) by rat adherent peritoneal cells, but they induced lower levels of production of NO than Escherichia coli LPS. The participation of the inducible isoform of NO synthase (iNOS) was confirmed by the finding of an increased expression of both iNOS mRNA and iNOS protein. These observations might help to explain (i) the acute outcome of Brucella infection in rodents...

  2. Surface exposure of O1 serotype lipopolysaccharide in Klebsiella pneumoniae strains expressing different K antigens.

    OpenAIRE

    J. M. Tomás; Camprubi, S; Merino, S.; Davey, M. R.; P. Williams

    1991-01-01

    Surface exposure of the O1 serotype lipopolysaccharide in encapsulated Klebsiella pneumoniae strains belonging to different serotypes was examined by using the O1 antigen-specific bacteriophages FC3-1 and FC3-2 in conjunction with immunogold electron microscopy and enzyme immunoassays with specific antisera. Despite the presence of the capsular polysaccharide, the O1 antigen was exposed at the cell surface in strains producing K2, K7, K8, K12, K19, K21, K22, K34, K35, K42, K45, K55, K57, K62,...

  3. Molecular Dynamics and NMR Spectroscopy Studies of E. coli Lipopolysaccharide Structure and Dynamics

    OpenAIRE

    Wu, Emilia l; Engstro?m, Olof; Jo, Sunhwan; Stuhlsatz, Danielle; Yeom, Min sun; Klauda, Jeffery b; Widmalm, Göran; Im, Wonpil

    2013-01-01

    Lipopolysaccharide (LPS), a component of Gram-negative bacterial outer membranes, comprises three regions: lipid A, core oligosaccharide, and O-antigen polysaccharide. Using the CHARMM36 lipid and carbohydrate force fields, we have constructed a model of an Escherichia coli R1 (core) O6 (antigen) LPS molecule. Several all-atom bilayers are built and simulated with lipid A only (LIPA) and varying lengths of 0 (LPS0), 5 (LPS5), and 10 (LPS10) O6 antigen repeating units; a single unit of O6 anti...

  4. Induction of inflammatory cytokines in bovine alveolar macrophages following stimulation with Pasteurella haemolytica lipopolysaccharide.

    OpenAIRE

    Yoo, H. S.; Maheswaran, S. K.; Lin, G.; Townsend, E. L.; Ames, T. R.

    1995-01-01

    Bovine tumor necrosis factor alpha (TNF-alpha) and interleukin-1 beta (IL-1 beta) cDNAs were generated by reverse transcription and then by PCR amplification from lipopolysaccharide (LPS)-stimulated alveolar macrophage RNA. The amplified cDNAs were cloned into pPow and expressed in Escherichia coli DH5 alpha. The expressed proteins were confirmed as TNF-alpha and IL-1 beta by Western blot (immunoblot) analysis and bioassays. We then used the cloned genes as probes in Northern (RNA) blots and ...

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

    Energy Technology Data Exchange (ETDEWEB)

    Straatsma, TP

    2006-12-01

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

  6. Lipopolysaccharide Extracellular Emulsifier Produced by Penicillium citrinum

    OpenAIRE

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

    2006-01-01

    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 emulsifyin...

  7. Lipopolysaccharide and silica-stimulated mononuclear cell prostaglandin production in ulcerative colitis

    OpenAIRE

    Punchard, Neville A.; John Cason; Jonathan Mullins; Chaman Chander; Thompson, Richard P. H.

    2000-01-01

    Basal, lipopolysaccharide (LPS) and silica-stimulated prostaglandin (PG) production were compared between peripheral blood mononuclear cells (PBMNC) from UC patients and healthy subjects (HS). Basal and LPS-stimulated PBMNC PGI2, but not PGE2, production was greater in UC. LPS stimulated both PGE2 and PGI2 by PBMNC from HS and UC patients. Silica stimulated production of both PGs by cells from HS but only PGE2 by cells from UC patients. The differences in responses to silica and LPS may resul...

  8. Lipopolysaccharides of Pseudomonas spp. that stimulate plant growth: composition and use for strain identification.

    OpenAIRE

    Weger, L. A.; Jann, B.; Jann, K.; Lugtenberg, B.

    1987-01-01

    The outer membrane proteins of a series of fluorescent, root-colonizing, plant-growth-stimulating Pseudomonas spp. having been characterized (L. A. de Weger et al., J. Bacteriol. 165:585-594, 1986), the lipopolysaccharides (LPSs) of these strains were examined. The chemical composition of the LPSs of the three best-studied plant-growth-stimulating Pseudomonas strains WCS358, WCS361, and WCS374 and of P. aeruginosa PAO1 as a reference strain was determined and appeared to differ from strain to...

  9. Noradrenaline inhibits lipopolysaccharide-induced tumor necrosis factor and interleukin 6 production in human whole blood.

    OpenAIRE

    Poll, T.; Jansen, J.; Endert, E.; Sauerwein, H. P.; Deventer, S. J.

    1994-01-01

    Sepsis and lipopolysaccharide (LPS) trigger the systemic release of both cytokines and catecholamines. Cytokines are known to be capable of eliciting a stress hormone response in vivo. The present study sought insight into the effect of noradrenaline on LPS-induced release of tumor necrosis factor alpha (TNF) and interleukin 6 (IL-6) in human whole blood. Whole blood was incubated with LPS for 4 h at 37 degrees C in the presence and absence of noradrenaline and/or specific alpha and beta anta...

  10. Astragalus mongholicus polysaccharide inhibits lipopolysaccharide-induced production of TNF-? and interleukin-8

    OpenAIRE

    Yuan Yuan, Mei Sun

    2009-01-01

    AIM: To explore the effect of Astragalus mongholicus polysaccharide (APS) on gene expression and mitogen-activated protein kinase (MAPK) transcriptional activity in intestinal epithelial cells (IEC).METHODS: IEC were divided into control group, lipopolysaccharide (LPS) group, LPS+ 50 ?g/mL APS group, LPS+ 100 ?g/mL APS group, LPS+ 200 ?g/mL APS group, and LPS+ 500 ?g/mL APS group. Levels of mRNAs in LPS-induced inflammatory factors, tumor necrosis factor (TNF)-? and interleukin (IL)-8, w...

  11. A protein secretion system linked to bacteroidete gliding motility and pathogenesis

    OpenAIRE

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

    2009-01-01

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

  12. Lipopolysaccharide-Induced Middle Ear Inflammation Disrupts the cochlear Intra-Strial Fluid–Blood Barrier through Down-Regulation of Tight Junction Proteins

    Science.gov (United States)

    Zhang, Jinhui; Chen, Songlin; Hou, Zhiqiang; Cai, Jing; Dong, Mingmin; Shi, Xiaorui

    2015-01-01

    Middle ear infection (or inflammation) is the most common pathological condition that causes fluid to accumulate in the middle ear, disrupting cochlear homeostasis. Lipopolysaccharide, a product of bacteriolysis, activates macrophages and causes release of inflammatory cytokines. Many studies have shown that lipopolysaccharides cause functional and structural changes in the inner ear similar to that of inflammation. However, it is specifically not known how lipopolysaccharides affect the blood-labyrinth barrier in the stria vascularis (intra-strial fluid–blood barrier), nor what the underlying mechanisms are. In this study, we used a cell culture-based in vitro model and animal-based in vivo model, combined with immunohistochemistry and a vascular leakage assay, to investigate lipopolysaccharide effects on the integrity of the mouse intra-strial fluid–blood barrier. Our results show lipopolysaccharide-induced local infection significantly affects intra-strial fluid–blood barrier component cells. Pericytes and perivascular-resident macrophage-like melanocytes are particularly affected, and the morphological and functional changes in these cells are accompanied by substantial changes in barrier integrity. Significant vascular leakage is found in the lipopolysaccharide treated-animals. Consistent with the findings from the in vivo animal model, the permeability of the endothelial cell monolayer to FITC-albumin was significantly higher in the lipopolysaccharide-treated monolayer than in an untreated endothelial cell monolayer. Further study has shown the lipopolysaccharide-induced inflammation to have a major effect on the expression of tight junctions in the blood barrier. Lipopolysaccharide was also shown to cause high frequency hearing loss, corroborated by previous reports from other laboratories. Our findings show lipopolysaccharide-evoked middle ear infection disrupts inner ear fluid balance, and its particular effects on the intra-strial fluid–blood barrier, essential for cochlear homeostasis. The barrier is degraded as the expression of tight junction-associated proteins such as zona occludens 1, occludin, and vascular endothelial cadherin are down-regulated. PMID:25815897

  13. Involvement of Semicarbazide-Sensitive Amine Oxidase-Mediated Deamination in Lipopolysaccharide-Induced Pulmonary Inflammation

    Science.gov (United States)

    Yu, Peter H.; Lu, Li-Xin; Fan, Hui; Kazachkov, Mychaylo; Jiang, Zhong-Jian; Jalkanen, Sirpa; Stolen, Craig

    2006-01-01

    Semicarbazide-sensitive amine oxidase (SSAO) resides on the vascular endothelium and smooth muscle cell surface and is capable of deaminating short chain aliphatic amines and producing toxic aldehydes and hydrogen peroxide. The enzyme, also known as a vascular adhesion protein-1, is involved in the inflammation process. This intriguing protein with dual functions is increased in the serum of diabetic and heart failure patients. In the present study we assessed the involvement of SSAO in a lipopolysaccharide-induced pulmonary inflammation model using transgenic mice that overexpress human vascular adhesion protein-1. Overexpression of SSAO activity increased the formation of protein-formaldehyde deposits in tissues. Lysine residues of proteins were the primary targets for cross-linkage with formaldehyde derived from deamination of methylamine. Lipopolysaccharide-induced increases in inflammatory cells in the bronchoalveolar lavage (BAL) fluid were significantly higher in the transgenic than in the nontransgenic mice. BAL cell counts were also higher in the untreated transgenic than in nontransgenic mice. Blocking SSAO activity with a selective inhibitor significantly reduced the number of neutrophils as well as levels of macrophage inflammatory protein-1?, granulocyte colony-stimulating factor, tumor necrosis factor-?, and interleukin-6 in the BAL fluid. Inhalation of methylamine also increased BAL neutrophil counts. Together, these results suggest a role for SSAO-mediated deamination in pulmonary inflammation. PMID:16507887

  14. Anti-inflammatory activity and mechanism of surfactin in lipopolysaccharide-activated macrophages.

    Science.gov (United States)

    Zhang, Yuanyuan; Liu, Chuan; Dong, Bin; Ma, Xiaolei; Hou, Lihua; Cao, Xiaohong; Wang, Chunling

    2015-04-01

    Surfactin is primarily produced by Bacillus natto TK-1 and is one of the most powerful biosurfactants. It consists of a heptapeptide interlinked with a ?-hydroxy fatty acid. Because of its special structure, surfactin shows broad biological effects, including anti-tumour, anti-microbial and anti-mycoplasma activities. It also has potential anti-inflammatory activity; however, the anti-inflammatory mechanism of surfactin has not been explored. In this study, we investigated the anti-inflammatory mechanism of surfactin in lipopolysaccharide (LPS)-stimulated macrophages. Surfactin exhibited an anti-inflammatory effect without cytotoxicity at certain concentrations, and the lipopolysaccharide (LPS)-stimulated cells appeared normal after surfactin treatment. Surfactin significantly inhibited the increased expression of IFN-?, IL-6, iNOS and nitric oxide (NO). TLR4 is the critical receptor for LPS; therefore, the TLR4 signal transduction pathway is the primary pathway that mediates LPS-induced inflammation. The results show that surfactin downregulated the LPS-induced TLR4 protein expression of macrophages and indicated that the surfactin-mediated signal pathway was involved in with TLR4. The subsequent studies demonstrated that surfactin exhibited anti-inflammatory effects by attenuating the activation of nuclear factor-?B (NF-?B), which is involved in the nuclear factor-?B (NF-?B) cell signalling pathways. These results suggest that surfactin may be a new therapeutic agent for inflammation. PMID:25331175

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

    Scientific Electronic Library Online (English)

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

    2014-05-01

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

  16. 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 ... cDermott EM, O'Neill ... LA. Immunology. 2004 Oct;113(2):153-62. (.png) (.s ... l-like receptor-4. Authors Palsson-McDermott EM, O'Neill ... LA. Publication Immunology. 2004 Oct;113(2):153-62 ...

  17. The dilution rate affects the outer membrane protein and lipopolysaccharide composition of Haemophilus influenzae type b grown under iron limitation.

    OpenAIRE

    Langford, P R; Moxon, E R

    1993-01-01

    When Haemophilus influenzae type b was grown under iron limitation in continuous culture, the dilution rate affected the outer membrane protein and lipopolysaccharide composition. Investigations of the effect of the reduced availability of iron or other environmental parameters on these surface components should be controlled for growth rate.

  18. Recognition of individual strains of fast-growing rhizobia by using profiles of membrane proteins and lipopolysaccharides.

    OpenAIRE

    Maagd, R.; Rossum, C.; Lugtenberg, B. J.

    1988-01-01

    Membrane protein and lipopolysaccharide profiles of Rhizobium leguminosarum (biovars viciae, trifolii, and phaseoli), R. meliloti, and Agrobacterium tumefaciens strains were analyzed and compared by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Differences in one or both profiles allowed us to distinguish all 18 R. leguminosarum strains tested in this study from each other.

  19. Cloning and characterization of the Escherichia coli Heptosyltransferase III: Exploring substrate specificity in lipopolysaccharide core biosynthesis.

    Science.gov (United States)

    Mudapaka, Jagadesh; Taylor, Erika Anne

    2015-06-01

    Bacterial lipopolysaccharide (LPS) molecules are an important cell surface component that enables adhesion to surfaces and cell motility, amongst other functions. In Escherichia coli, there are multiple Heptosyltransferase enzymes involved in the biosynthesis of the core region of LPS. Here we describe the first ever cloning, expression, purification and characterization of Heptosyltransferase III (HepIII) from E. coli, which catalyzes the addition of an l-glycero-d-manno-heptose (Hep) residue to the growing LPS core via an ?(1?7) bond. Inspired by results from our lab on the E. coli HepI, we assessed the catalytic efficiency with phospho-Hep2-Kdo2-Lipid A (PH2K2LA) and two deacylated analogues. PMID:25957775

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2010-04-01

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

  1. The effect of Pseudomonas syringae pv. atrofaciens 9417 lipopolysaccharide on mutagenicity in pro- and eukaryotic systems

    Directory of Open Access Journals (Sweden)

    Gvozdyak R. I.

    2010-02-01

    Full Text Available Aim. To study the effect of lipopolysaccharide (LPS of Pseudomonas syringae pv. atrofaciens on spontaneous and induced mutations in pro- and eukaryotic test-systems. Methods. Mutagenic and antimutagenic properties of LPS were studied in Allium cepa-test and Ames test. Results. LPS does not influence the spontaneous mutations of Salmonella typhimurium and decreases the level of mutations induced by potassium dichromate and N-methyl-N'-nitro-N'-nitrosoguanidine. LPS reduces a mitotic index at concentrations of 10.0 and 5.0 mg/ml and increases the number of chromosomes’ fragments in cells of A. cepa root apical meristem at concentrations of 5.0 and 2.5 mg/ml. Conclusion. Different effect of LPS on mutagenesis in pro- and eukaryotic cells has been established. LPS revealed mutagenic properties in A. cepa-test and antimutagenic properties in Ames test.

  2. Characterization of Lipopolysaccharides of Bradyrhizobium japonicum KDR 15 Heavy Metal Tolerant

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    ALFI DATIN ZAUQIAH

    2006-09-01

    Full Text Available The lipopolysaccharide (LPS of Bradyrhizobium japonicum KDR 15 heavy metal tolerant strain was isolated by miniphenol-water extraction and yielded LPS in phenol and water phase. The LPS KDR 15 was further characterized by sodium dodecyl sulfate-polyacrilamide gel electrophoresis (SDS PAGE and showed many bands distributed from an area of high until low molecular weight (LPS IA, IB, and II. Composition analysis of the LPS had been done after acetic acid 1% hydrolysis. The polysaccharide portion consist of glucose, sucrose, galactose, mannose, xylose, arabinose, rhamnose, ribose, glucosamine, and 3-deoksi-D-manno-oktulosonat (KDO. Lipid A portion consisted of C16:0 and C18:1. The LPS also contained 0.02% of protein and 1.7% of phosphate. The presence of functional groups that shows negative charge densities such as phosphate and carboxyl within LPS KDR 15 assumed to be a potentially binding sites for accumulating heavy metals.

  3. Differences in lipopolysaccharide-induced signaling between conventional dendritic cells and macrophages.

    Science.gov (United States)

    Zanoni, Ivan; Granucci, Francesca

    2010-01-01

    Dendritic cells (DCs) and macrophages contribute to the activation of immune responses against infectious agents. They sense the presence of microbes through germline-encoded pattern-recognition receptors (PRRs), which recognize pathogen-associated molecular patterns (PAMPs). Among the different PAMPs, the response to lipopolysaccharide (LPS) is one of the best characterized. Upon LPS encounter DCs undergo an activation process and acquire the ability to prime both natural killer and T-cell responses after migration to lymph nodes. Once they completed the effector phase, DCs reach a terminal differentiation stage and eventually die by apoptosis. By contrast, macrophages do not leave the tissue upon LPS recognition. They first initiate inflammatory processes and then switch to an anti-inflammatory phenotype to restore tissue homeostasis. In this review we will focus on the molecular bases of the divergent responses of DCs and macrophages to LPS. PMID:20579765

  4. Lipopolysaccharide-binding protein as a major plasma protein responsible for endotoxemic shock.

    Science.gov (United States)

    Gallay, P; Heumann, D; Le Roy, D; Barras, C; Glauser, M P

    1993-01-01

    Because lipopolysaccharide (LPS)-binding protein (LBP) sensitizes monocytes to LPS in vitro, it has been suggested that LBP initiates host defenses against Gram-negative bacteria. The role of LBP in vivo, and particularly in endotoxemic shock, is unknown, however. Therefore an IgG against murine LBP was prepared. It was found to neutralize binding of LPS and subsequent activation of murine macrophages in vitro. This anti-LBP protected mice against the lethal effect of LPS when given at the same time as LPS challenge, but it failed to protect mice when delayed 15 min after LPS challenge. The same preparation was also effective after challenge with lipid A but not after challenge with Staphylococcus aureus enterotoxin. The protection was correlated with a strong decrease of circulating tumor necrosis factor. These data demonstrate that in vivo LBP is a major mediator of the lethal effects of endotoxemia. Images Fig. 1 PMID:7694297

  5. A lectin S-domain receptor kinase mediates lipopolysaccharide sensing in Arabidopsis thaliana.

    Science.gov (United States)

    Ranf, Stefanie; Gisch, Nicolas; Schäffer, Milena; Illig, Tina; Westphal, Lore; Knirel, Yuriy A; Sánchez-Carballo, Patricia M; Zähringer, Ulrich; Hückelhoven, Ralph; Lee, Justin; Scheel, Dierk

    2015-04-01

    The sensing of microbe-associated molecular patterns (MAMPs) triggers innate immunity in animals and plants. Lipopolysaccharide (LPS) from Gram-negative bacteria is a potent MAMP for mammals, with the lipid A moiety activating proinflammatory responses via Toll-like receptor 4 (TLR4). Here we found that the plant Arabidopsis thaliana specifically sensed LPS of Pseudomonas and Xanthomonas. We isolated LPS-insensitive mutants defective in the bulb-type lectin S-domain-1 receptor-like kinase LORE (SD1-29), which were hypersusceptible to infection with Pseudomonas syringae. Targeted chemical degradation of LPS from Pseudomonas species suggested that LORE detected mainly the lipid A moiety of LPS. LORE conferred sensitivity to LPS onto tobacco after transient expression, which demonstrated a key function in LPS sensing and indicated the possibility of engineering resistance to bacteria in crop species. PMID:25729922

  6. Hepatoprotective and antibacterial activity of Lippia nodiflora Linn. against lipopolysaccharides on HepG2 cells

    Science.gov (United States)

    Arumanayagam, S.; Arunmani, M.

    2015-01-01

    Background: Lippia nodiflora (LN) Linn is a small herb distributed throughout the world. The plant extracts of LN is used traditionally as an analgesic, anti-inflammatory, antioxidant, antibacterial, antimicrobial, antipyretic, antitumor, antidiabetic, and possess hepatoprotective properties. Materials and Methods: To study the antibacterial and hepatoprotective effect of LN, we used methanolic extracts of leaves on HepG2 cells. Lipopolysaccharides (LPS) is a well-characterized hepatotoxin, so toxicity was induced on liver cells using LPS. Up-regulation of inflammation genes were quantified. Results and Conclusions: In our present study, we have showed that LN reduced reactive oxygen species (ROS) production against LPS induced toxicity on HepG2 cells, and ther by decreased the apoptotic gene expression and protect the liver cells against toxicity. PMID:25709206

  7. Bacteriophage adhesin-coated long-period gratings for bacterial lipopolysaccharide recognition

    Science.gov (United States)

    Koba, Marcin; ?mietana, Mateusz; Brzozowska, Ewa; Górska, Sabina; Mikulic, Predrag; Bock, Wojtek J.

    2014-05-01

    In this work we report an application of the optical fiber long-period gratings (LPGs) working near the dispersion turning point of higher order cladding modes for bacterial lipopolysaccharide (LPS) recognition. We show that when the LPG is functionalized with the bacteriophage adhesin, it is capable of very specific LPS detection. Thus, we compare label-free binding effect for specific to the adhesin LPS-positive and non-specific LPS-negative. The resonance wavelength shift induced by the LPS-positive reaches 2.9 nm, while for LPS-negative the shift is negligible. The LPG-based sensing structure allows for monitoring of the binding phenomenon in real time and with good accuracy.

  8. Lipophilic antioxidants prevent lipopolysaccharide-induced mitochondrial dysfunction through mitochondrial biogenesis improvement.

    Science.gov (United States)

    Bullón, Pedro; Román-Malo, Lourdes; Marín-Aguilar, Fabiola; Alvarez-Suarez, José Miguel; Giampieri, Francesca; Battino, Maurizio; Cordero, Mario D

    2015-01-01

    Oxidative stress is implicated in several infectious diseases. In this regard, lipopolysaccharide (LPS), an endotoxic component, induces mitochondrial dysfunction and oxidative stress in several pathological events such as periodontal disease or sepsis. In our experiments, LPS-treated fibroblasts provoked increased oxidative stress, mitochondrial dysfunction, reduced oxygen consumption and mitochondrial biogenesis. After comparing coenzyme Q10 (CoQ10) and N-acetylcysteine (NAC), we observed a more significant protection of CoQ10 than of NAC, which was comparable with other lipophilic and hydrophilic antioxidants such as vitamin E or BHA respectively. CoQ10 improved mitochondrial biogenesis by activating PGC-1? and TFAM. This lipophilic antioxidant protection was observed in mice after LPS injection. These results show that mitochondria-targeted lipophilic antioxidants could be a possible specific therapeutic strategy in pharmacology in the treatment of infectious diseases and their complications. PMID:25447593

  9. Functional Identification of Proteus mirabilis eptC Gene Encoding a Core Lipopolysaccharide Phosphoethanolamine Transferase

    Science.gov (United States)

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

    2014-01-01

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

  10. Study of Nitric Oxide production by murine peritoneal macrophages induced by Brucella Lipopolysaccharide

    Directory of Open Access Journals (Sweden)

    Kavoosi G

    2001-07-01

    Full Text Available Brueclla is a gram negative bacteria that causes Brucellosis. Lipopolysaccharide (LPS ", the pathogenic agent of Brucella is composed of O-chain, core oligosaccharide and lipid A. in addition, the structural and biological properties of different LPS extracted from different strains are not identical. The first defense system against LPS is nonspecific immunity that causes macrophage activation. Activated macrophages produce oxygen and nitrogen radicals that enhance the protection against intracellular pathogens.In this experiment LPS was extracted by hot phenol- water procedure and the effect of various LPSs on nitric oxide prodution by peritoneal mouse macrophages was examined.Our results demonstrated that the effect of LPS on nitric oxide production is concentration-dependent we observed the maximum response in concentration of 10-20 microgram per milliliter. Also our results demonstrate that LPS extracted from vaccine Brucella abortus (S 19 had a highe effect on nitric oxide production than the LPS from other strains

  11. Antigenic potential of a highly conserved Neisseria meningitidis lipopolysaccharide inner core structure defined by chemical synthesis.

    Science.gov (United States)

    Reinhardt, Anika; Yang, You; Claus, Heike; Pereira, Claney L; Cox, Andrew D; Vogel, Ulrich; Anish, Chakkumkal; Seeberger, Peter H

    2015-01-22

    Neisseria meningitidis is a leading cause of bacterial meningitis worldwide. We studied the potential of synthetic lipopolysaccharide (LPS) inner core structures as broadly protective antigens against N. meningitidis. Based on the specific reactivity of human serum antibodies to synthetic LPS cores, we selected a highly conserved LPS core tetrasaccharide as a promising antigen. This LPS inner core tetrasaccharide induced a robust IgG response in mice when formulated as an immunogenic glycoconjugate. Binding of raised mouse serum to a broad collection of N. meningitidis strains demonstrated the accessibility of the LPS core on viable bacteria. The distal trisaccharide was identified as the crucial epitope, whereas the proximal Kdo moiety was immunodominant and induced mainly nonprotective antibodies that are responsible for lack of functional protection in polyclonal serum. Our results identified key antigenic determinants of LPS core glycan and, hence, may aid the design of a broadly protective immunization against N. meningitidis. PMID:25601073

  12. [Study of the binding between bacterial lipopolysaccharides and polymyxin by a bioluminescent method].

    Science.gov (United States)

    Katsev, A M; Onuchina, I G; Gordienko, A I; Beloglazov, V A

    2002-01-01

    For rating the interaction of lipopolysaccharides (LPS) with polymixin B (PmB) a bacterial bioluminescence is offered to be used. Bioluminescence level of bacteria decreases under free antibiotic amounts action. It is shown, that as a result of interaction with LPS antibiotic properties of PmB are reduced, and the intensity of bacterial bioluminescence is restored. The bioluminescence level in such system characterizes the LPS quantity. Kinetic properties of bacterial light emission at the presence of LPS and PmB are investigated as well as equilibrium state of the system. Kinetic and equilibrium constants describing this reaction are determined. The conditions of quantitative bioluminescent definition of LPS in an interval of concentration 0.166-10 micrograms/ml have been chosen and calibration curves are presented. PMID:12924022

  13. Attenuation of lipopolysaccharide (LPS-induced cytotoxicity by tocopherols and tocotrienols

    Directory of Open Access Journals (Sweden)

    Keiko Nishio

    2013-01-01

    Full Text Available Lipopolysaccharide (LPS induces host inflammatory responses and tissue injury and has been implicated in the pathogenesis of various age-related diseases such as acute respiratory distress syndrome, vascular diseases, and periodontal disease. Antioxidants, particularly vitamin E, have been shown to suppress oxidative stress induced by LPS, but the previous studies with different vitamin E isoforms gave inconsistent results. In the present study, the protective effects of ?- and ?-tocopherols and ?- and ?-tocotrienols on the oxidative stress induced by LPS against human lung carcinoma A549 cells were studied. They suppressed intracellular reactive oxygen formation, lipid peroxidation, induction of inflammatory mediator cytokines, and cell death. Tocopherols were incorporated into cultured cells much slower than tocotrienols but could suppress LPS-induced oxidative stress at much lower intracellular concentration than tocotrienols. Considering the bioavailability, it was concluded that ?-tocopherol may exhibit the highest protective capacity among the vitamin E isoforms against LPS-induced oxidative stress.

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

    Directory of Open Access Journals (Sweden)

    Abdella E

    2008-11-01

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

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2012-11-13

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

  16. Lipopolysaccharide binding protein enhances the responsiveness of alveolar macrophages to bacterial lipopolysaccharide. Implications for cytokine production in normal and injured lungs.

    Science.gov (United States)

    Martin, T R; Mathison, J C; Tobias, P S; Letúrcq, D J; Moriarty, A M; Maunder, R J; Ulevitch, R J

    1992-12-01

    A plasma lipopolysaccharide (LPS)-binding protein (LBP) has been shown to regulate the response of rabbit peritoneal macrophages and human blood monocytes to endotoxin (LPS). We investigated whether LBP is present in lung fluids and the effects of LBP on the response of lung macrophages to LPS. Immunoreactive LBP was detectable in the lavage fluids of patients with the adult respiratory distress syndrome by immunoprecipitation followed by Western blotting, and also by specific immunoassay. In rabbits, the LBP appeared to originate outside of the lungs, inasmuch as mRNA transcripts for LBP were identified in total cellular RNA from liver, but not from lung homogenates or alveolar macrophages. Purified LBP enhanced the response of human and rabbit alveolar macrophages to both smooth form LPS (Escherichia coli O111B:4) and rough form LPS (Salmonella minnesota Re595). In the presence of LBP and LPS, the onset of tumor necrosis factor-alpha (TNF alpha) production occurred earlier and at an LPS threshold dose that was as much as 1,000-fold lower for both types of LPS. In rabbit alveolar macrophages treated with LBP and LPS, TNF alpha mRNA appeared earlier, reached higher levels, and had a prolonged half-life as compared with LPS treatment alone. Neither LPS nor LPS and LBP affected pHi or [Cai++] in alveolar macrophages. Specific monoclonal antibodies to CD14, a receptor that binds LPS/LBP complexes, inhibited TNF alpha production by human alveolar macrophages stimulated with LPS alone or with LPS/LBP complexes, indicating the importance of CD14 in mediating the effects of LPS on alveolar macrophages. Thus, immunoreactive LBP accumulates in lung lavage fluids in patients with lung injury and enhances LPS-stimulated TNF alpha gene expression in alveolar macrophages by a pathway that depends on the CD14 receptor. LBP may play an important role in augmenting TNF alpha expression by alveolar macrophages within the lungs. PMID:1281827

  17. Measurement of immunoglobulin M, immunoglobulin G, and immunoglobulin A antibodies against Yersinia enterocolitica by enzyme-linked immunosorbent assay: comparison of lipopolysaccharide and whole bacterium as antigen.

    OpenAIRE

    Granfors, K.; Viljanen, M. K.; Toivanen, A.

    1981-01-01

    An enzyme-linked immunosorbent assay (ELISA) for the detection and quantitation of human immunoglobulin M (IgM), IgG, and IgA antibodies against Yersinia enterocolitica by using lipopolysaccharides as antigens is described. The results obtained with the lipopolysaccharide ELISA were compared with the results of the whole bacterium ELISA. The correlations observed were good for each immunoglobulin class. Cross-reactions between Y. enterocolitica serotypes O:3 and O:9, Yersinia pseudotuberculos...

  18. Lipopolysaccharide-Binding Protein- and CD14-Dependent Activation of Mitogen-Activated Protein Kinase p38 by Lipopolysaccharide in Human Neutrophils Is Associated with Priming of Respiratory Burst

    OpenAIRE

    Yan, Sen Rong; Al-Hertani, Walla; Byers, David; Bortolussi, Robert

    2002-01-01

    Neutrophil (PMN) functions can be primed for greatly increased oxidative radical release by exposure to certain agents such as lipopolysaccharide (LPS). Although a variety of signaling pathways involving both tyrosine kinases and mitogen-activated protein (MAP) kinases may be operative, the mechanisms of PMN priming are still not understood. We found that PMN priming was not achieved by treatment of cells with a very low concentration (5 ng/ml) of LPS unless additional “helper” factors we...

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

    Directory of Open Access Journals (Sweden)

    Fábio Gonçalves

    2009-12-01

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

  20. The detection of eight putative periodontal pathogens in adult and rapidly progressive periodontitis patients: An institutional study

    OpenAIRE

    Joshi Vinayak; Vandana K

    2007-01-01

    Purpose: Periodontal disease is a commonly prevalent problem faced alike by both the developed and third world countries but showing wide variations in prevalence and severity across different geographical areas. The purpose was to identify Actinobacillus actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Ekinella corrodens (Ec), Campylobacter rectus (Cr), Bacteroides forsythus (Bf), Treponema denticola (Td) and Fusobacterium nucleatum (Fn)...

  1. Efficacy of the de novo-derived antimicrobial peptide WLBU2 against oral bacteria.

    Science.gov (United States)

    Novak, Karen F; Diamond, William J; Kirakodu, Sreenatha; Peyyala, Rebecca; Anderson, Kimberly W; Montelaro, Ronald C; Mietzner, Timothy A

    2007-05-01

    The efficacy of a novel synthetic antimicrobial peptide (WLBU2) was evaluated against three oral microorganisms (grown planktonically): Streptococcus gordonii, Fusobacterium nucleatum, and Porphyromonas gingivalis. WLBU2 killed all three species, with F. nucleatum being the most susceptible. WLBU2 also reduced the bacterial burden of S. gordonii and F. nucleatum biofilms. PMID:17325219

  2. Efficacy of the De Novo-Derived Antimicrobial Peptide WLBU2 against Oral Bacteria?

    OpenAIRE

    Novak, Karen F.; Diamond, William J.; Kirakodu, Sreenatha; Peyyala, Rebecca; Anderson, Kimberly W.; Montelaro, Ronald C.; Mietzner, Timothy A.

    2007-01-01

    The efficacy of a novel synthetic antimicrobial peptide (WLBU2) was evaluated against three oral microorganisms (grown planktonically): Streptococcus gordonii, Fusobacterium nucleatum, and Porphyromonas gingivalis. WLBU2 killed all three species, with F. nucleatum being the most susceptible. WLBU2 also reduced the bacterial burden of S. gordonii and F. nucleatum biofilms.

  3. Efficacy of the De Novo-Derived Antimicrobial Peptide WLBU2 against Oral Bacteria?

    Science.gov (United States)

    Novak, Karen F.; Diamond, William J.; Kirakodu, Sreenatha; Peyyala, Rebecca; Anderson, Kimberly W.; Montelaro, Ronald C.; Mietzner, Timothy A.

    2007-01-01

    The efficacy of a novel synthetic antimicrobial peptide (WLBU2) was evaluated against three oral microorganisms (grown planktonically): Streptococcus gordonii, Fusobacterium nucleatum, and Porphyromonas gingivalis. WLBU2 killed all three species, with F. nucleatum being the most susceptible. WLBU2 also reduced the bacterial burden of S. gordonii and F. nucleatum biofilms. PMID:17325219

  4. Use of Quantitative PCR and Culture Methods To Characterize Ecological Flux in Bacterial Biofilms?

    OpenAIRE

    Dalwai, F.; Spratt, D. A.; Pratten, J.

    2007-01-01

    An in vitro model of supragingival plaque associated with gingivitis was characterized by traditional culture techniques, comparative 16S rRNA gene sequencing of isolates, and quantitative PCR (QPCR). Actinomyces naeslundii, Prevotella spp., and Porphyromonas gingivalis increased under conditions emulating gingivitis. Gram-negative species and total bacteria were dramatically underestimated by culture compared to the estimates obtained by QPCR.

  5. Changes in the incidence of periodontal pathogens during long-term monitoring and after application of antibacterial drugs.

    Czech Academy of Sciences Publication Activity Database

    Janatová, T.; Najmanová, Lucie; Neubauerová, Lenka; Kyselková, Martina; Novotná, Gabriela; Spížek, Jaroslav; Janata, Ji?í; Dušková, J.

    2009-01-01

    Ro?. 54, ?. 5 (2009), s. 429-435. ISSN 0015-5632 R&D Projects: GA MŠk 1M06011; GA MZd NR9119 Institutional research plan: CEZ:AV0Z50200510 Keywords : DUAL-SPECIES BIOFILMS * ACTINOBACILLUS-ACTINOMYCETEMCOMITANS * PORPHYROMONAS-GINGIVALIS Subject RIV: EE - Microbiology, Virology Impact factor: 0.978, year: 2009

  6. Keratocan and Lumican Regulate Neutrophil Infiltration and Corneal Clarity in Lipopolysaccharide-induced Keratitis by Direct Interaction with CXCL1*

    OpenAIRE

    Carlson, Eric C; Lin, Michelle; Liu, Chia-Yang; Kao, Winston W-Y; Perez, Victor L.; Pearlman, Eric

    2007-01-01

    Keratocan and lumican are keratan-sulfate proteoglycans (KSPG), which have a critical role in maintaining corneal clarity. To determine whether these KSPGs have a role in corneal inflammation, we examined Kera?/? and Lum?/? mice in a model of lipopolysaccharide (LPS)-induced keratitis in which wild-type mice develop increased corneal thickness and haze due to neutrophil infiltration to the corneal stroma. Corneal thickness increases caused by LPS mice were significantly lower in Kera?...

  7. Tumor necrosis factor-alpha mRNA remains unstable and hypoadenylated upon stimulation of macrophages by lipopolysaccharides.

    OpenAIRE

    Mijatovic, Tatjana; Houzet, Laurent; Defrance, Patrick; Droogmans, Louis; Huez, Georges; Kruys, Ve?ronique

    2000-01-01

    TNF-alpha gene expression is regulated at transcriptional and post-transcriptional levels in mouse macrophages. The post-transcriptional regulation is mediated by the AU-rich element (ARE) located in the TNF-alpha mRNA 3' untranslated region (UTR), which controls its translation and stability. In resting macrophages, the ARE represses TNF-alpha mRNA translation. Activation of macrophages with various agents [for example lipopolysaccharide (LPS), viruses] results in translational derepression,...

  8. Antimicrobial Action and Cell Agglutination by the Eosinophil Cationic Protein Are Modulated by the Cell Wall Lipopolysaccharide Structure

    OpenAIRE

    Pulido, David; Moussaoui, Mohammed; Andreu, David; Nogués, M. Victòria; Torrent, Marc; Boix, Ester

    2012-01-01

    Antimicrobial proteins and peptides (AMPs) are essential effectors of innate immunity, acting as a first line of defense against bacterial infections. Many AMPs exhibit high affinity for cell wall structures such as lipopolysaccharide (LPS), a potent endotoxin able to induce sepsis. Hence, understanding how AMPs can interact with and neutralize LPS endotoxin is of special relevance for human health. Eosinophil cationic protein (ECP) is an eosinophil secreted protein with high activity against...

  9. The immunocytochemical localization of tumour necrosis factor and leukotriene in the rat kidney after treatment with lipopolysaccharide.

    OpenAIRE

    Kita, T.; Tanaka, N.; Nagano, T.

    1993-01-01

    Abundant inflammatory cells such as polymorphonuclear leucocytes and macrophages accumulated and adhered to the endothelial surface of glomerular and intertubular veins and capillaries in rat kidneys after administration of bacterial lipopolysaccharide (LPS). There was also damage to both endothelial cells and proximal tubular cells, including intracytoplasmic oedema, and an increase in the number of lysosomes in the proximal tubular cells in the LPS-treated samples. Immunocytochemistry was u...

  10. Comparison of Inflammation, Organ Damage, and Oxidant Stress Induced by Salmonella enterica Serovar Muenchen Flagellin and Serovar Enteritidis Lipopolysaccharide

    OpenAIRE

    Liaudet, Lucas; Murthy, Kanneganti G. K.; Mabley, Jon G.; Pacher, Pa?l; Soriano, Francisco G.; Salzman, Andrew L.; Szabo?, Csaba

    2002-01-01

    Gram-negative sepsis is related to the activation of interconnected inflammatory cascades in response to bacteria and their products. Recent work showed that flagellin, the monomeric subunit of bacterial flagella, triggers innate immune responses mediated by Toll-like receptor 5. Here, we compared the effects of Salmonella enterica serovar Enteritidis lipopolysaccharide (LPS) and recombinant Salmonella enterica serovar Muenchen flagellin administered intravenously (100 ?g) to mice. Flagellin...

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

    OpenAIRE

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

    1987-01-01

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

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

    OpenAIRE

    Elass, Elisabeth; Masson, Maryse; Mazurier, Joël; Legrand, Dominique

    2002-01-01

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

  13. Pivotal Role of Reactive Oxygen Species in Differential Regulation of Lipopolysaccharide-Induced Prostaglandins Production in Macrophages

    OpenAIRE

    Zhao, Guiqing; Yu, Rui; Deng, Jing; Zhao, Qiong; Li, Yongchao; Joo, Myungsoo; van Breemen, Richard B.; John W. Christman; Xiao, Lei

    2013-01-01

    Gram-negative bacterial endotoxin lipopolysaccharide (LPS) triggers the production of inflammatory cytokines, reactive oxygen species (ROS), and prostaglandins (PGs) by pulmonary macrophages. Here, we investigated if ROS influenced PGs production in response to LPS treatment in mouse bone marrow-derived macrophages (BMDM). We observed that pretreatment of BMDM with two structurally unrelated ROS scavengers, MnTMPyP and EUK-134, not only prevented LPS-induced ROS accumulation, but also attenua...

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

    OpenAIRE

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

    2011-01-01

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

  15. Dendritic cell loss from nonlymphoid tissues after systemic administration of lipopolysaccharide, tumor necrosis factor, and interleukin 1

    OpenAIRE

    1995-01-01

    Dendritic cells (DC) in nonlymphoid organs can internalize and process foreign antigens before migrating to secondary lymphoid tissues to initiate primary immune responses. However, there is little information on which stimuli promote migration of DC from the tissues. Systemic administration of lipopolysaccharide (LPS), which induces in vivo production of cytokines, led to a reduction in the numbers of major histocompatibility complex class II-positive (Ia+) leukocytes in mouse hearts and kid...

  16. The effect of nebivolol on the production of nitric oxide induced by bacterial lipopolysaccharide and peptidoglycan in mice

    OpenAIRE

    Fadi El-Rami; Hampartsoum Barsoumian; Joseph Simaan; Alexander M. Abdelnoor

    2010-01-01

    Nitric oxide (NO) plays a pivotal role in main- taining balance of physiological events in many systems including the autonomic, cardiovas- cular, hematological, and pulmonary systems. Lipopolysaccharide (LPS) and peptidoglycan (PGN), components of the outer cell membranes of Gram-negative bacteria and cell walls of Gram-positive bacteria respectively, are in- criminated in NO-induced septic shock. Ne- bivolol is a third generation ?1- adrenoceptor blocker with a vasodilatory property attribu...

  17. Lipopolysaccharide-induced interleukin 8 production by human whole blood is enhanced by epinephrine and inhibited by hydrocortisone.

    OpenAIRE

    Poll, T.; Lowry, S. F.

    1997-01-01

    To determine the effect of epinephrine and hydrocortisone on lipopolysaccharide (LPS)-induced interleukin 8 (IL-8) production, human whole blood was stimulated with LPS in the presence or absence of these stress hormones. Epinephrine caused a dose-dependent increase in LPS-induced IL-8 production, which was mediated exclusively via beta-adrenergic receptors, as reflected by the facts that beta (but not alpha) receptor blockade reversed the epinephrine effect and beta (but not alpha) receptor ...

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

    OpenAIRE

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

    1990-01-01

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

  19. Lipid IVA inhibits synthesis and release of tumor necrosis factor induced by lipopolysaccharide in human whole blood ex vivo

    OpenAIRE

    1990-01-01

    Tumor necrosis factor (TNF) released by lipopolysaccharide (LPS)- stimulated mononuclear phagocytes is a critical mediator of sepsis. We examined the capacities of rough mutant Salmonella typhimurium LPS (Rc) and LPS partial structures lipid A, monophosphoryl lipid A (MPLA), lipid IVA, and lipid X to induce production of TNF in whole blood. Rc LPS (0.0001-10 ng/ml) produced a dose-dependent release of TNF as determined by cytotoxicity of actinomycin D-sensitized L929 murine fibroblasts. Lipid...

  20. The inhibitory effect of quercitrin gallate on iNOS expression induced by lipopolysaccharide in Balb/c mice

    OpenAIRE

    Jo, Hyun-ye; Kim, Youngsoo; Nam, Sang-yoon; Lee, Beom Jun; Kim, Yun-bae; Yun, Young Won; Ahn, Byeongwoo

    2008-01-01

    Quercetin 3-O-?-(2"-galloyl)-rhamnopyranoside (QGR) is a naturally occurring quercitrin gallate, which is a polyphenolic compound that was originally isolated from Persicaria lapathifolia (Polygonaceae). QGR has been shown to have an inhibitory effect on nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated macrophage RAW 264.7 cells. Therefore, this study was conducted to investigate the inhibitory effect of QGR on nitric oxide production and inducible nitric oxide synthases (...

  1. Enhancement of Methacholine-Evoked Tracheal Contraction Induced by Bacterial Lipopolysaccharides Depends on Epithelium and Tumor Necrosis Factor

    OpenAIRE

    Secher, T.; Rodrigues Coelho, F.; Noulin, N.; Lino Dos Santos Franco, A.; Quesniaux, V.; Lignon, J.; Mitchell, J.; Moser, R.; Gomes, E.; Mirotti, L.; Tavares-de-lima, W.; Ryffel, B.; Boris Vargaftig, B.; Russo, M.

    2012-01-01

    Inhaled bacterial lipopolysaccharides (LPSs) induce an acute tumour necrosis factor-alpha (TNF-?-) dependent inflammatory response in the murine airways mediated by Toll-like receptor 4 (TLR4) via the myeloid differentiation MyD88 adaptor protein pathway. However, the contractile response of the bronchial smooth muscle and the role of endogenous TNF? in this process have been elusive. We determined the in vivo respiratory pattern of C57BL/6 mice after intranasal LPS administration with or w...

  2. Enterobacter agglomerans lipopolysaccharide-induced changes in pulmonary surfactant as a factor in the pathogenesis of byssinosis.

    OpenAIRE

    Delucca, A. J.; Brogden, K. A.; Engen, R.

    1988-01-01

    Lipopolysaccharide (LPS) from Enterobacter agglomerans and pulmonary surfactant mixtures were centrifuged in discontinuous sucrose gradients to determine whether LPS bound to surfactant and examined in a Langmuir trough with a Wilhelmy balance to determine whether LPS altered the surface activity of surfactant. The LPS was found to bind to the surfactant and altered its surface tension properties. The binding of LPS to surfactant in the lung may change the physiological properties of surfacta...

  3. Role of Lipopolysaccharide Phase Variation in Susceptibility of Haemophilus influenzae to Bactericidal Immunoglobulin M Antibodies in Rabbit Sera

    OpenAIRE

    Erwin, Alice L.; Brewah, Yambasu A.; Couchenour, Debra A.; Barren, Philip R.; Burke, Stephen J.; Choi, Gil H.; Lathigra, Raju; Hanson, Mark S.; Weiser, Jeffrey N.

    2000-01-01

    The effect of phase variation of lipopolysaccharide (LPS) structure on the susceptibility of Haemophilus influenzae to complement-dependent killing by normal human sera and normal rat sera has been described previously. The phase-variable structure phosphorylcholine (ChoP) confers susceptibility to human serum, since ChoP on the bacterial cell surface binds to serum C-reactive protein and activates complement. In contrast, expression of gal?1,4gal, a second phase-variable epitope that is als...

  4. Effects of repeated intratracheally administered lipopolysaccharide on primary and secondary specific antibody responses and on body weight gain of broilers

    OpenAIRE

    Lai, T. L. H.; Nieuwland, M. G. B.; Kemp, B.; Aarnink, A. J. A.; Parmentier, H. K.

    2011-01-01

    Earlier, we reported that pathogen-associated molecular patterns such as lipopolysaccharide (LPS), when administered intratracheally (i.t.), affected primary and secondary specific antibody responses to antigens administered concurrently, either i.t. or systemically, and also affected BW gain (BWG) of layers and broilers. In the present study, we evaluated the effects of repeated i.t. challenge with LPS concurrently with or before i.t. immunizations with the specific antigens human serum albu...

  5. Monoclonal antibodies specific for Escherichia coli J5 lipopolysaccharide: cross-reaction with other gram-negative bacterial species.

    OpenAIRE

    Mutharia, L. M.; Crockford, G.; Bogard, W. C.; Hancock, R. E.

    1984-01-01

    Four monoclonal antibodies against Escherichia coli J5 were studied. Each of these monoclonal antibodies reacted with purified lipopolysaccharides from E. coli J5, the deep rough mutant Salmonella minnesota Re595, Agrobacterium tumefaciens, and Pseudomonas aeruginosa PAO1 as well as with the purified lipid A of P. aeruginosa. Enzyme-linked immunosorbent assays using the outer membranes from a variety of gram-negative bacteria demonstrated that these lipid A-specific monoclonal antibodies inte...

  6. Mutants of Ralstonia (Pseudomonas) solanacearum sensitive to antimicrobial peptides are altered in their lipopolysaccharide structure and are avirulent in tobacco.

    OpenAIRE

    Titarenko, Elena; Lopez Solanilla, Emilia; García Olmedo, Francisco; Rodriguez Palenzuela, Pablo

    1997-01-01

    Ralstonia solanacearum K60 was mutagenized with the transposon Tn5, and two mutants, M2 and M88, were isolated. Both mutants were selected based on their increased sensitivity to thionins, and they had the Tn5 insertion in the same gene, 34 bp apart. Sequence analysis of the interrupted gene showed clear homology with the rfaF gene from Escherichia coli and Salmonella typhimurium (66% similarity), which encodes a heptosyltransferase involved in the synthesis of the lipopolysaccharide (LPS) co...

  7. Ginsenoside Rg3 Alleviates Lipopolysaccharide-Induced Learning and Memory Impairments by Anti-Inflammatory Activity in Rats

    OpenAIRE

    Lee, Bombi; Sur, Bongjun; Park, Jinhee; Kim, Sung-Hun; Kwon, Sunoh; Yeom, Mijung; Shim, Insop; Lee, Hyejung; Hahm, Dae-Hyun

    2013-01-01

    The purpose of this study was to examine whether ginsenoside Rg3 (GRg3) could improve learning and memory impairments and inflammatory reactions induced by injecting lipopolysaccharide (LPS) into the brains of rats. The effects of GRg3 on proinflammatory mediators in the hippocampus and the underlying mechanisms of these effects were also investigated. Injection of LPS into the lateral ventricle caused chronic inflammation and produced deficits in learning in a memory-impairment animal model....

  8. Inhibitory Effect of Astragalus Polysaccharides on Lipopolysaccharide-Induced TNF-a and IL-1? Production in THP-1 Cells

    OpenAIRE

    Aiping Lu; Cheng Lu; Li Xu; Jun Shu; Xiaojuan He

    2012-01-01

    Astragalus polysaccharides (APS), one of main bioactive components in Astragalus membranaceus Bunge, has been reported to possess anti-inflammatory activities, but the molecular mechanisms behind this activity are largely unknown. This study aimed to investigate expression of inflammatory cytokines and the MAPK/NF-?B pathway in human THP-1 macrophages induced by lipopolysaccharide (LPS). The results showed that the concentrations of TNF-a and IL-1? released from LPS stimulated THP-1 cells i...

  9. A low-level diode laser therapy reduces the lipopolysaccharide (LPS)-induced periodontal ligament cell inflammation

    International Nuclear Information System (INIS)

    The purpose of this study was to investigate the cytologic effects of inflammatory periodontal ligament cells in vitro after low-level laser therapy. Human periodontal ligament cells were cultured, exposed to lipopolysaccharide and subjected to low-level laser treatment of 5?J?cm?2 or 10?J?cm?2 using a 920?nm diode laser. A periodontal ligament cell attachment was observed under a microscope, and the cell viability was quantified by a mitochondrial colorimetric assay. Lipopolysaccharide-treated periodontal ligament cells were irradiated with the low-level laser, and the expression levels of several inflammatory markers, iNOS, TNF-? and IL-1, and pErk kinase, were analyzed by reverse transcription polymerase chain reaction and western blot. The data were collected and analyzed by one-way analysis of variance; p < 0.05 indicated a statistically significant difference. The low-level laser treatment of periodontal ligament cells increased their ability to attach and survive. After irradiation, the expression levels of iNOS, TNF-? and IL-1 in lipopolysaccharide-exposed periodontal ligament cells decreased over time (p < 0.05). In periodontal ligament cells, low-level diode laser treatment increased the cells’ proliferative ability and decreased the expression of the examined inflammatory mediators. (letters)

  10. A low-level diode laser therapy reduces the lipopolysaccharide (LPS)-induced periodontal ligament cell inflammation

    Science.gov (United States)

    Huang, T. H.; Chen, C. C.; Liu, S. L.; Lu, Y. C.; Kao, C. T.

    2014-07-01

    The purpose of this study was to investigate the cytologic effects of inflammatory periodontal ligament cells in vitro after low-level laser therapy. Human periodontal ligament cells were cultured, exposed to lipopolysaccharide and subjected to low-level laser treatment of 5?J?cm?2 or 10?J?cm?2 using a 920?nm diode laser. A periodontal ligament cell attachment was observed under a microscope, and the cell viability was quantified by a mitochondrial colorimetric assay. Lipopolysaccharide-treated periodontal ligament cells were irradiated with the low-level laser, and the expression levels of several inflammatory markers, iNOS, TNF-? and IL-1, and pErk kinase, were analyzed by reverse transcription polymerase chain reaction and western blot. The data were collected and analyzed by one-way analysis of variance; p periodontal ligament cells increased their ability to attach and survive. After irradiation, the expression levels of iNOS, TNF-? and IL-1 in lipopolysaccharide-exposed periodontal ligament cells decreased over time (p periodontal ligament cells, low-level diode laser treatment increased the cells’ proliferative ability and decreased the expression of the examined inflammatory mediators.

  11. Lactoferrin-lipopolysaccharide interaction: involvement of the 28-34 loop region of human lactoferrin in the high-affinity binding to Escherichia coli 055B5 lipopolysaccharide.

    Science.gov (United States)

    Elass-Rochard, E; Roseanu, A; Legrand, D; Trif, M; Salmon, V; Motas, C; Montreuil, J; Spik, G

    1995-12-15

    The ability of lactoferrin (Lf), an iron-binding glycoprotein that is also called lactotransferrin, to bind lipopolysaccharide (LPS) may be relevant to some of its biological properties. A knowledge of the LPS-binding site on Lf may help to explain the mechanism of its involvement in host defence. Our report reveals the presence of two Escherichia coli 055B5 LPS-binding sites on human Lf (hLf): a high-affinity binding site (Kd 3.6 +/- 1 nM) and a low-affinity binding site (Kd 390 +/- 20 nM). Bovine Lf (bLf), which shares about 70% amino acid sequence identity with hLf, exhibits the same behaviour towards LPS. Like hLf, bLf also contains a low- and a high-affinity LPS-binding site. The Kd value (4.5 +/- 2 nM) corresponding to the high-affinity binding site is similar to that obtained for hLf. Different LPS-binding sites for human serum transferrin have been suggested, as this protein, which is known to bind bacterial endotoxin, produced only 12% inhibition of hLf-LPS interaction. Binding and competitive binding experiments performed with the N-tryptic fragment (residues 4-283), the C-tryptic fragment (residues 284-692) and the N2-glycopeptide (residues 91-255) isolated from hLf have demonstrated that the high-affinity binding site is located in the N-terminal domain I of hLf, and the low-affinity binding site is present in the C-terminal lobe. The inhibition of hLf-LPS interaction by a synthetic octadecapeptide corresponding to residues 20-37 of hLf and lactoferricin B (residues 17-41), a proteolytic fragment from bLf, revealed the importance of the 28-34 loop region of hLf and the homologous region of bLf for LPS binding. Direct evidence that this amino acid sequence is involved in the high-affinity binding to LPS was demonstrated by assays carried out with EGS-loop hLf, a recombinant hLf mutated at residues 28-34. PMID:8554529

  12. Up-regulation of microglial cathepsin C expression and activity in lipopolysaccharide-induced neuroinflammation

    Directory of Open Access Journals (Sweden)

    Fan Kai

    2012-05-01

    Full Text Available Abstract Background Cathepsin C (Cat C functions as a central coordinator for activation of many serine proteases in inflammatory cells. It has been recognized that Cat C is responsible for neutrophil recruitment and production of chemokines and cytokines in many inflammatory diseases. However, Cat C expression and its functional role in the brain under normal conditions or in neuroinflammatory processes remain unclear. Our previous study showed that Cat C promoted the progress of brain demyelination in cuprizone-treated mice. The present study further investigated the Cat C expression and activity in lipopolysaccharide (LPS-induced neuroinflammation in vivo and in vitro. Methods C57BL/6?J mice were intraperitoneally injected with either 0.9% saline or lipopolysaccharide (LPS, 5?mg/kg. Immunohistochemistry (IHC and in situ hybridization (ISH were used to analyze microglial activation, TNF-?, IL-1?, IL-6, iNOS mRNAs expressions and cellular localization of Cat C in the brain. Nitrite assay was used to examine microglial activation in vitro; RT-PCR and ELISA were used to determine the expression and release of Cat C. Cat C activity was analyzed by cellular Cat C assay kit. Data were evaluated for statistical significance with paired t test. Results Cat C was predominantly expressed in hippocampal CA2 neurons in C57BL/6?J mice under normal conditions. Six hours after LPS injection, Cat C expression was detected in cerebral cortical neurons; whereas, twenty-four hours later, Cat C expression was captured in activated microglial cells throughout the entire brain. The duration of induced Cat C expression in neurons and in microglial cells was ten days and three days, respectively. In vitro, LPS, IL-1? and IL-6 treatments increased microglial Cat C expression in a dose-dependent manner and upregulated Cat C secretion and its activity. Conclusions Taken together, these data indicate that LPS and proinflammatory cytokines IL-1?, IL-6 induce the expression, release and upregulate enzymatic activity of Cat C in microglial cells. Further investigation is required to determine the functional role of Cat C in the progression of neuroinflammation, which may have implications for therapeutics for the prevention of neuroinflammation-involved neurological disorders in the future.

  13. Influence of lipopolysaccharide-binding protein on pulmonary inflammation in gram-negative pneumonia.

    Science.gov (United States)

    Taddonio, Michael A; Dolgachev, Vladislav; Bosmann, Markus; Ward, Peter A; Su, Grace; Wang, Stewart C; Hemmila, Mark R

    2015-06-01

    Lipopolysaccharide-binding protein (LBP) is upregulated as part of the acute-phase response. Lipopolysaccharide-binding protein has a known multifunctional role in potentiating the recognition, clearance, and killing of gram-negative bacteria. In a Klebsiella pneumonia model, we previously demonstrated that LBP gene-deficient mice (LBP) mice experience increased mortality when compared with wild-type (Wt) mice (98% vs. 59%). We hypothesize that LBP is essential to bacterial clearance from the lung, and its absence leads to alteration of the pulmonary inflammatory response to pneumonia. Twelve- to 16-week-old female C57Bl/6 Wt mice and age-matched LBP mice were administered 1 × 10 colony-forming units of Klebsiella pneumoniae by intratracheal injection. Animals were euthanized at 6, 12, 24, or 36 h after inoculation. Lung tissue and bronchoalveolar lavage samples were obtained. Lung homogenate samples were assayed to determine quantitative bacterial load per whole lung, proinflammatory cytokine concentrations, myeloperoxidase activity, and assessment of pulmonary leukocyte populations. In vitro production of inflammatory mediators were also assayed after LPS stimulation of peritoneal macrophages isolated from Wt, Toll-like receptor 4 (TLR4)-deficient, and LBP mice. The LBP mice demonstrated significantly elevated levels of bacteria in the lung at 24 and 36 h when compared with Wt controls. The average lung levels of proinflammatory cytokines interleukin-1? (IL-1?), IL-6, keratinocyte-derived chemokine, and macrophage-inflammatory protein-2 were greater in the LBP mice and remained elevated longer when compared with those in the Wt mice. Myeloperoxidase activity, an indicator of neutrophil content, was significantly increased at time 36 h in the LBP mice. After in vitro stimulation of peritoneal macrophages with LPS, production of IL-1?, IL-6, IL-10, keratinocyte-derived chemokine, and macrophage-inflammatory protein-1? were suppressed in LBP and TLR4-deficient mice compared with that in Wt. Absence of a functional LBP gene results in diminished clearance of gram-negative bacteria from the pulmonary system. Failure to recognize and clear gram-negative bacteria via the LBP/TLR4 axis results in an initial delayed inflammatory response. This delay in LBP mice is followed by excessive amplification and prolonged elevation of proinflammatory mediators and neutrophil sequestration within the lungs. PMID:25643011

  14. Keratinocyte growth factor (KGF)-1 and -2 protein and gene expression in human gingival fibroblasts.

    Science.gov (United States)

    Sanale, Ali-Reza; Firth, James D; Uitto, Veli-Jukka; Putnins, Edward E

    2002-02-01

    The onset and progression of periodontal disease is associated with significant changes in the epithelial component of the attachment complex. From the early to the advanced stages of periodontal disease increased epithelial cell proliferation, migration and invasion into the surrounding connective tissue takes place. Concomitantly there is a significant increase in proinflammatory cytokine expression in periodontal tissue and quantitative and qualitative changes in the subgingival microflora, including an increase in gram-negative microorganisms. One of the most significant virulence factors of these bacteria is lipopolysaccharide (LPS) connected to the outer membrane. Two important growth factors controlling epithelial behavior are Keratinocyte Growth Factor-1 (KGF-1) and -2 (KGF-2). Connective tissue cells express these growth factors, but only epithelial cells respond to them. We studied the effect of proinflammatory cytokines and LPS on gingival fibroblast expression of KGF-1 and KGF-2 in vitro. Gingival fibroblasts were found to express KGF-1 and -2 in culture but only KGF-1 protein and gene expression was stimulated by serum, in a concentration-dependent manner by proinflammatory cytokines IL-1alpha, IL-1beta, TNF-alpha and IL-6 and LPS isolated from Porphyromonas gingivalis and Escherichia coli. The local increase in proinflammatory cytokine expression and the accumulation of LPS in disease sites may therefore stimulate gingival fibroblast expression of KGF-1. We hypothesize that this local increase in KGF-1 expression may, via a paracrine mechanism, stimulate local epithelial cell proliferation, migration and invasion during the onset and progression of periodontitis. PMID:11842940

  15. Resistance of MMP9 and TIMP1 to endotoxin tolerance.

    Science.gov (United States)

    Muthukuru, Manoj; Cutler, Christopher W

    2015-07-01

    Inflammatory cytokines activate tissue collagenases such as matrix metalloproteinases (MMPs). MMPs are antagonized by tissue inhibitors of metalloproteinases (TIMPs) that attempt to regulate excessive collagenase activity during inflammatory conditions. During chronic inflammatory conditions, induction of endotoxin tolerance negatively regulates the cytokine response in an attempt to curtail excessive host tissue damage. However, little is known about how downregulation of inflammatory cytokines during endotoxin tolerance regulates MMP activities. In this study, human monocyte-derived macrophages were either sensitized or further challenged to induce tolerance with lipopolysaccharide (LPS) from Porphyromonas gingivalis (PgLPS) or Escherichia coli (EcLPS). Inflammatory cytokines, such as TNF-? and IL-1?, and levels of MMP9 and TIMP1 were analyzed by a combination of cytometric bead array, western blot/gelatin zymography and real-time RT-PCR. Functional blocking with anti-TLR4 but not with anti-TLR2 significantly downregulated TNF-? and IL-1?. However, MMP9 levels were not inhibited by toll-like receptor (TLR) blocking. Interestingly, endotoxin tolerance significantly upregulated TIMP1 relative to MMP9 and downmodulated MMP9 secretion and its enzymatic activity. These results suggest that regulatory mechanisms such as induction of endotoxin tolerance could inhibit MMP activities and could facilitate restoring host tissue homeostasis. PMID:25951835

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

    DEFF Research Database (Denmark)

    Ma, Zhiwei; Dagnaes-Hansen, Frederik

    2015-01-01

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

  17. Molecular cloning and functional characterization of a mouse gene upregulated by lipopolysaccharide treatment reveals alternative splicing

    International Nuclear Information System (INIS)

    Treatment of mouse cells with lipopolysaccharide (LPS) potently initiates an inflammatory response, but the underlying mechanisms are unclear. We therefore sought to characterize cDNA sequences of a new mouse LPS-responsive gene, and to evaluate the effects of MLrg. Full-length cDNAs were obtained from LPS-treated NIH3T3 cells. We report that the MLrg gene produces two alternative splice products (GenBank Accession Nos. (DQ316984) and (DQ320011)), respectively, encoding MLrgW and MLrgS polypeptides. Both proteins contain zinc finger and leucine zipper domains and are thus potential regulators of transcription. Expression of MLrgW and MLrgS were robustly upregulated following LPS treatment, and the proteins were localized predominantly in the nuclear membrane and cytoplasm. In stable transfectants over-expressing MLrgW the proportion of cells in G1 phase was significantly reduced, while in cells over-expressing MLrgS the proportion of cells in G2 was significantly increased; both proteins are thus potential regulators of cell cycle progression. Upregulation of MLrgW and MLrgS may be an important component of the LPS inflammatory pathway and of the host response to infection with GNB.

  18. Effect of low-intensity electromagnetic radiation on structurization properties of bacterial lipopolysaccharide

    Directory of Open Access Journals (Sweden)

    Grigory E. Brill

    2014-09-01

    Full Text Available Purpose — to investigate the effects of low-intensity electromagnetic radiation on the process of dehydration selforganization of bacterial lipopolysaccharide (LPS. Material and Methods — The method of wedge dehydration has been used to study the structure formation of bacterial LPS. Image-phases analysis included their qualitative characteristics, as well as the calculation of quantitative indicators, followed by statistical analysis. Results — Low-intensity ultra high frequency (UHF radiation (1 GHz, 0.1 ?W/cm2, 10 min has led to the changes in the suspension system of the LPS-saline reflected in the kinetics of structure formation. Conclusion — 1 GHz corresponds to the natural frequency of oscillation of water clusters and, presumably, the effect of UHF on structure of LPS mediates through the changes in water-salt environment. Under these conditions, properties of water molecules of hydration and possibly the properties of hydrophobic and hydrophilic regions in the molecule of LPS, which can affect the ability of toxin molecules to form aggregates change. Therefore the LPS structure modification may result in the change of its toxic properties.

  19. Depression of afferent arc of the in vivo cytotoxic T-cell immunity by bacterial lipopolysaccharides

    International Nuclear Information System (INIS)

    The afferent arc of the in vivo cytotoxic T-cell immunity assessed by second set rejection of ascitic allogeneic tumors was shown to be depressed by bacterial lipopolysaccharide (LPS) that was administered simultaneously with or 1 day before injection of allogeneic spleen cells as stimulators. Two different LPSs from Escherichia coli O55 and Klebsiella O3 displayed similar activities whereas dextran sulfate, concanavalin A, or poly A:U was not effective. Stimulator activities of allogeneic cells was not directly modified by LPS. Any definite suppressor activity on afferent or efferent arc of the T-cell response was not demonstrable in mice receiving LPS and allogeneic cells. Further, the LPS effect for immune depression was not diminished by whole body X-ray irradiation to the recipient at 300 R, which ablated the B-cell reactivity to LPS for polyclonal activation, or by treatment of the recipient with carrageenan, a known toxic agent to macrophages. It was suggested from these results that LPS suppresses the cytotoxic T-cell immunity by modulating responder T cells to be temporarily refractory to the allogeneic stimulus rather than by activating suppressor cells such as radiation-sensitive lymphocytes and carrageenan-sensitive macrophages

  20. Astragalus mongholicus polysaccharide inhibits lipopolysaccharide-induced production of TNF-? and interleukin-8

    Directory of Open Access Journals (Sweden)

    Yuan Yuan, Mei Sun, Ke-Shen Li

    2009-08-01

    Full Text Available AIM: To explore the effect of Astragalus mongholicus polysaccharide (APS on gene expression and mitogen-activated protein kinase (MAPK transcriptional activity in intestinal epithelial cells (IEC.METHODS: IEC were divided into control group, lipopolysaccharide (LPS group, LPS+ 50 ?g/mL APS group, LPS+ 100 ?g/mL APS group, LPS+ 200 ?g/mL APS group, and LPS+ 500 ?g/mL APS group. Levels of mRNAs in LPS-induced inflammatory factors, tumor necrosis factor (TNF-? and interleukin (IL-8, were measured by reverse transcription-polymerase chain reaction. MAPK protein level was measured by Western blotting.RESULTS: The levels of TNF-? and IL-8 mRNAs were significantly higher in IEC with LPS-induced damage than in control cells. APS significantly abrogated the LPS-induced expression of the TNF-? and IL-8 genes. APS did not block the activation of extracellular signal-regulated kinase or c Jun amino-terminal kinase, but inhibited the activation of p38, suggesting that APS inhibits LPS-induced production of TNF-? and IL-8 mRNAs, possibly by suppressing the p38 signaling pathway.CONCLUSION: APS-modulated bacterial product-mediated p38 signaling represents an attractive strategy for prevention and treatment of intestinal inflammation.

  1. A Distal Locus Element Mediates IFN-? Priming of Lipopolysaccharide-Stimulated TNF Gene Expression

    Directory of Open Access Journals (Sweden)

    Nancy A. Chow

    2014-12-01

    Full Text Available Interferon ? (IFN-? priming sensitizes monocytes and macrophages to lipopolysaccharide (LPS stimulation, resulting in augmented expression of a set of genes including TNF. Here, we demonstrate that IFN-? priming of LPS-stimulated TNF transcription requires a distal TNF/LT locus element 8 kb upstream of the TNF transcription start site (hHS-8. IFN-? stimulation leads to increased DNase I accessibility of hHS-8 and its recruitment of interferon regulatory factor 1 (IRF1, and subsequent LPS stimulation enhances H3K27 acetylation and induces enhancer RNA synthesis at hHS-8. Ablation of IRF1 or targeting the hHS-8 IRF1 binding site in vivo with Cas9 linked to the KRAB repressive domain abolishes IFN-? priming, but does not affect LPS induction of the gene. Thus, IFN-? poises a distal enhancer in the TNF/LT locus by chromatin remodeling and IRF1 recruitment, which then drives enhanced TNF gene expression in response to a secondary toll-like receptor (TLR stimulus.

  2. Neonatal lipopolysaccharide exposure induces sexually dimorphic sickness behavior in adult rats

    Scientific Electronic Library Online (English)

    Maria M., Bernardi; Lívia P., Teixeira; Ana P., Ligeiro-de-Oliveira; Wothan, Tavares-de-Lima; João, Palermo-Neto; Thiago B., Kirsten.

    2014-06-01

    Full Text Available The aim of the present study was to evaluate whether neonatal exposure to lipopolysaccharide (LPS; 50 µg/kg, i.p., on postnatal day 2) induces depressive- and/or anxiety-like effects and sexually dimorphic responses in rats challenged with LPS (100 µg/kg, i.p.) in adulthood. The results revealed tha [...] t males presented a less depressive state in the forced swim test and exhibited no changes in general motor activity in the open field test. Females exhibited an increase in sickness behavior, revealing different behavioral strategies in response to a bacterial disease. The male rats also exhibited higher cell proliferation, reflected by bone marrow and peripheral blood counts, and female rats exhibited a decrease in corticosterone levels. No changes were observed in the elevated plus maze or peripheral cytokine levels (interleukin-1? and tumor necrosis factor-?). Neonatal exposure to LPS induced sexually dimorphic behavioral, neuroendocrine, and immune effects after an LPS challenge in adulthood, differentially affecting male and female susceptibility to disease later in life.

  3. Dietary Astragalus polysaccharide alleviated immunological stress in broilers exposed to lipopolysaccharide.

    Science.gov (United States)

    Liu, Lei; Shen, Jing; Zhao, Chao; Wang, Xiaofei; Yao, Junhu; Gong, Yuesheng; Yang, Xiaojun

    2015-01-01

    This study was conducted to investigate whether dietary Astragalus polysaccharide (APS) could alleviate immunological stress response of chickens after challenge with lipopolysaccharide (LPS). A total of 360 one-day-old commercial Arbor Acres broilers were randomly assigned in a 2 × 2 factorial design. The main factors were immunological stress (LPS or saline) and dietary APS (0 or 3g APS/kg feed). At 12, 14, 33 and 35 days of age, chickens were injected intramuscularly with either 500 ?g/kg body weight of LPS or sterile saline. The results showed that the decreased daily feed intake and daily weight gain caused by immunological stress were dramatically attenuated by APS supplementation. The LPS challenge led to an increased mRNA abundance of TLR4, NF-?B, IL-1?, IL-6, avian uncoupling protein, ?1-acid glycoprotein, hemopexin and y(+)LAT2. However, these negative effects of the LPS administration were ameliorated by APS supplementation. Moreover, dietary APS inhibited the LPS-induced depression of amino acid digestibilities. In conclusion, APS is able to alleviate LPS-induced immunological stress response in chickens. The beneficial effect may be attributed to suppressing the expression of pro-inflammatory cytokines through reducing the TLR4 and NF-?B genes transcription, and therewith improving energy and protein metabolism. PMID:25239195

  4. Occurrence of glycine in the core oligosaccharides of Hafnia alvei lipopolysaccharides-identification of disubstituted glycoform.

    Science.gov (United States)

    Gozdziewicz, Tomasz K; Man-Kupisinska, Aleksandra; Lugowski, Czes?aw; Lukasiewicz, Jolanta

    2015-05-18

    Endotoxins (lipopolysaccharides, LPS) are the main surface antigens and virulence factors of Gram-negative bacteria involved for example in the development of nosocomial infections and sepsis. They consist of three main regions: O-specific polysaccharide, core oligosaccharide, and lipid A. Bacteria modify LPS structure to escape the immune defence, but also to adapt to environmental conditions. LPS's structures are highly diversified in the O-specific polysaccharide region to evade bactericidal factors of immune system, but retain some common epitopes that are potential candidates for therapeutic strategies against bacterial infections. Common occurrence of glycine within the structure of LPS is a known phenomenon and was previously reported for variety of species. Since glycine residue substitutes mainly core oligosaccharide of LPS, especially inner core region, it was also considered as a part of common epitope for broad-reactive antimicrobial antibodies. Herein, we used multiple-stage electrospray ionisation mass spectrometry to identify glycine substitution in core oligosaccharide type characteristic for Hafnia alvei LPS, and isolated from five strains of different O-serotypes: 32, PCM 1190, PCM 1192, PCM 1200, and PCM 1209. The location of glycine in core oligosaccharide was determined in detail for LPS 1190 using ESI-MS(n). Three glycoforms were identified, including two mono-glycinylated and one diglycinylated core oligosaccharides. PMID:25541016

  5. Alginate microparticles loaded with lipopolysaccharide subunit antigen for mucosal vaccination against Klebsiella pneumoniae.

    Science.gov (United States)

    Jain, R R; Mehta, M R; Bannalikar, A R; Menon, M D

    2015-05-01

    Klebsiella pneumoniae (K. pneumoniae) is one of the commonest causes of nosocomial infections in human beings. Since K. pneumoniae infections are air borne type, controlling it by mucosal vaccination through nasal and pulmonary route could be a promising approach in order to simulate the natural infection. New vaccines such as subunit vaccines are safer than traditional vaccines, but they are less immunogenic. Therefore to enhance their immunogenicity, there is a need to develop potent and safe adjuvants and delivery systems. It has been established that micro-particles are one of the most potent adjuvants available for mucosal delivery of vaccines and they do so by improving uptake of encapsulated antigen by antigen presenting cells (APCs). Lipopolysaccharide (LPS), the antigenic fraction was extracted from K. pneumoniae by hot phenol extraction method. LPS loaded sodium alginate microparticles were prepared by emulsion ionic gelation method. Microparticles with particle size less than 5 ?m were obtained. Loading efficiency of the LPS loaded microparticles ranged from 76 to 82 %. Comparative in vivo immunogenicity studies were carried for free LPS and encapsulated LPS, administered via intramuscular, intratracheal and intranasal routes in Swiss albino mice. The study revealed that LPS encapsulated microparticles exhibit greater efficacy when administered by intra-tracheal route as compared to free LPS vaccine. PMID:25737397

  6. Sirtinol inhibits neutrophil elastase activity and attenuates lipopolysaccharide-mediated acute lung injury in mice.

    Science.gov (United States)

    Tsai, Yung-Fong; Yu, Huang-Ping; Chang, Wen-Yi; Liu, Fu-Chao; Huang, Zhen-Cheng; Hwang, Tsong-Long

    2015-01-01

    Enhanced activity of neutrophil elastase leads to a protease-antiprotease imbalance, and plays an essential pathogenic role in acute lung injury (ALI) and acute respiratory distress syndrome. We assayed the pharmacological effects and mechanisms of the action of sirtinol in human neutrophils, and in neutrophil elastase (HNE)-induced paw edema and lipopolysaccharide (LPS)-mediated ALI in mice. Sirtinol significantly inhibited the activity of HNE from human neutrophils in response to various stimulators. The inhibitory effects on HNE activity were not mediated through protein kinase A, calcium, extracellular-regulated kinase, c-Jun N-terminal kinase, p38 mitogen-activated protein kinase, Akt, or Src family kinases. Analysis of enzymatic activities showed that sirtinol inhibited HNE activity in a concentration-dependent manner. These results demonstrate that sirtinol does not affect neutrophil function and is an HNE inhibitor. In addition, administration of sirtinol significantly inhibited HNE-induced paw edema, and attenuated the myeloperoxidase activity and reduced pulmonary wet/dry weight ratio in the LPS-induced ALI mouse model. Our study indicates that sirtinol has anti-inflammatory effects through direct inhibition of HNE activity and attenuates HNE-induced and LPS-mediated tissue or organ injury in vivo. Sirtinol is a novel HNE inhibitor and may have the potential for clinical application in the treatment of inflammatory lung diseases. PMID:25666548

  7. Antioxidant properties of lutein contribute to the protection against lipopolysaccharide-induced uveitis in mice

    Directory of Open Access Journals (Sweden)

    Yao Xin-Sheng

    2011-10-01

    Full Text Available Abstract Background Lutein is an important eye-protective nutrient. This study investigates the protective effects and mechanisms of lutein on lipopolysaccharides (LPS-induced uveitis in mice. Methods Lutein, suspended in drinking water at a final concentration of 12.5 and 25 mg/mL, was administered to mice at 0.1 mL/10 g body weight for five consecutive days. Control and model group received drinking water only. Uveitis was induced by injecting LPS (100 mg per mouse into the footpad in the model and lutein groups on day 5 after the last drug administration. Eyes of the mice were collected 24 hours after the LPS injection for the detection of indicators using commercial kits and reverse transcription-polymerase chain reaction. Results LPS-induced uveitis was confirmed by significant pathological damage and increased the nitric oxide level in eye tissue of BALB/C mice 24 hours after the footpad injection. The elevated nitric oxide level was significantly reduced by oral administration of lutein (125 and 500 mg/kg/d for five days before LPS injection. Moreover, lutein decreased the malondialdehyde content, increased the oxygen radical absorbance capacity level, glutathione, the vitamin C contents and total superoxide dismutase (SOD and glutathione peroxidase (GPx activities. Lutein further increased expressions of copper-zinc SOD, manganese SOD and GPx mRNA. Conclusion The antioxidant properties of lutein contribute to the protection against LPS-induced uveitis, partially through the intervention of inflammation process.

  8. Proteome of monocyte priming by lipopolysaccharide, including changes in interleukin-1beta and leukocyte elastase inhibitor

    Directory of Open Access Journals (Sweden)

    Beranova-Giorgianni Sarka

    2008-05-01

    Full Text Available Abstract Background Monocytes can be primed in vitro by lipopolysaccharide (LPS for release of cytokines, for enhanced killing of cancer cells, and for enhanced release of microbicidal oxygen radicals like superoxide and peroxide. We investigated the proteins involved in regulating priming, using 2D gel proteomics. Results Monocytes from 4 normal donors were cultured for 16 h in chemically defined medium in Teflon bags ± LPS and ± 4-(2-aminoethyl-benzenesulfonyl fluoride (AEBSF, a serine protease inhibitor. LPS-primed monocytes released inflammatory cytokines, and produced increased amounts of superoxide. AEBSF blocked priming for enhanced superoxide, but did not affect cytokine release, showing that AEBSF was not toxic. After staining large-format 2D gels with Sypro ruby, we compared the monocyte proteome under the four conditions for each donor. We found 30 protein spots that differed significantly in response to LPS or AEBSF, and these proteins were identified by ion trap mass spectrometry. Conclusion We identified 19 separate proteins that changed in response to LPS or AEBSF, including ATP synthase, coagulation factor XIII, ferritin, coronin, HN ribonuclear proteins, integrin alpha IIb, pyruvate kinase, ras suppressor protein, superoxide dismutase, transketolase, tropomyosin, vimentin, and others. Interestingly, in response to LPS, precursor proteins for interleukin-1? appeared; and in response to AEBSF, there was an increase in elastase inhibitor. The increase in elastase inhibitor provides support for our hypothesis that priming requires an endogenous serine protease.

  9. Lipopolysaccharide increases gastric and circulating NUCB2/nesfatin-1 concentrations in rats

    Science.gov (United States)

    Stengel, Andreas; Goebel-Stengel, Miriam; Jawien, Janusz; Kobelt, Peter; Taché, Yvette; Lambrecht, Nils W.G.

    2014-01-01

    Bacterial lipopolysaccharide (LPS) is an established animal model to study the innate immune response to Gram-negative bacteria mimicking symptoms of infection including reduction of food intake. LPS decreases acyl ghrelin associated with decreased concentrations of circulating ghrelin-O-acyltransferase (GOAT) likely contributing to the anorexigenic effect. We also recently described the prominent expression of the novel anorexigenic hormone, nucleobindin2 (NUCB2)/nesfatin-1 in gastric X/A-like cells co-localized with ghrelin in different pools of vesicles. To investigate whether LPS would affect gastric and circulating NUCB2/nesfatin-1 concentration, ad libitum fed rats were equipped with an intravenous (iv) catheter. LPS was injected intraperitoneally (ip, 100 ?g/kg) and blood was withdrawn before and at 2, 5, 7 and 24 h post injection and processed for NUCB2/nesfatin-1 radioimmunoassay. Gastric corpus was collected to measure NUCB2 mRNA expression by RT-qPCR and NUCB2/nesfatin-1 protein concentration by Western blot. Injection of LPS increased plasma NUCB2/nesfatin-1 concentrations by 43%, 78% and 62% compared to vehicle at 2 h, 5 h and 7 h post injection respectively (p nesfatin-1 increase at 2 h was associated with increased corpus NUCB2 mRNA expression (p nesfatin-1 and ghrelin expression derived from the same cell by immune challenge. PMID:21782869

  10. PROTECTIVE EFFECTS OF MELATONIN AGAINST IDIOSYNCRATIC LIVER INJURY IN RATS CHALLENGED WITH CHLORPROMAZINE AND LIPOPOLYSACCHARIDE

    Directory of Open Access Journals (Sweden)

    Sulaiman Amal A

    2011-03-01

    Full Text Available The episode of inflammation during drug treatment predispose animals to tissue injury, raising the possibility that presence or absence of inflammation is an important susceptibility factor for drug toxicity in human , and this phenomenon is hypothesized to be related to idiosyncratic drug reactions. The present study was designed to investigate the possible protective effect of orally administered melatonin against hepatic injury in rats concurrently treated with chlorpromazine (CPZ and lipopolysaccharides (LPS. The protective effect of melatonin was studied through treatment of rats with single oral dose (10mg/kg, given seven days before and during the day of exposure to both CPZ and LPS. The animals were sacrificed 24 hrs post-challenge with LPS. Hepatic necrosis and cholestasis were assessed by measuring the activities of serum liver enzymes ALT, AST and ALP; in addition, total and direct bilirubin along with evaluation of histological alteration in hepatic tissue. The oxidative stress markers were assessed by measuring the levels of MDA and GSH in hepatic tissue homogenate. Analysis of data showed that melatonin supplementation attenuated markers of oxidative stress by reducing the levels of MDA and restoring GSH levels; melatonin also improved the observed biochemical and histological changes associated with exposure to CPZ and LPS. In conclusion, orally administered melatonin at pharmacological doses have promising protective effects against drug-induced idiosyncratic liver injury that may be of value in clinical practice.

  11. Gelam Honey Has a Protective Effect against Lipopolysaccharide (LPS-Induced Organ Failure

    Directory of Open Access Journals (Sweden)

    Mustafa Kassim

    2012-05-01

    Full Text Available Gelam honey exerts anti-inflammatory and antioxidant activities and is thought to have potent effects in reducing infections and healing wounds. The aim of this study was to investigate the effects of intravenously-injected Gelam honey in protecting organs from lethal doses of lipopolysaccharide (LPS. Six groups of rabbits (N = 6 were used in this study. Two groups acted as controls and received only saline and no LPS injections. For the test groups, 1 mL honey (500 mg/kg in saline was intravenously injected into two groups (treated, while saline (1 mL was injected into the other two groups (untreated; after 1 h, all four test groups were intravenously-injected with LPS (0.5 mg/kg. Eight hours after the LPS injection, blood and organs were collected from three groups (one from each treatment stream and blood parameters were measured and biochemical tests, histopathology, and myeloperoxidase assessment were performed. For survival rate tests, rabbits from the remaining three groups were monitored over a 2-week period. Treatment with honey showed protective effects on organs through the improvement of organ blood parameters, reduced infiltration of neutrophils, and decreased myeloperoxidase activity. Honey-treated rabbits also showed reduced mortality after LPS injection compared with untreated rabbits. Honey may have a therapeutic effect in protecting organs during inflammatory diseases.

  12. Interactions between magainin 2 and Salmonella typhimurium outer membranes: Effect of lipopolysaccharide structure

    International Nuclear Information System (INIS)

    The role of the outer membrane and lipopolysaccharide (LPS) in the interaction between the small cationic antimicrobial peptide magainin 2 and the Gram-negative cell envelope was studied by FT-IR spectroscopy. Magainin 2 alters the thermotropic properties of the outer membrane-peptidoglycan complexes from wild-type Salmonella typhimurium and a series of LPS mutants which display differential susceptibility to the bactericidal activity of cationic antibiotics. These results are correlated with the LPS phosphorylation pattern and charge (characterized by high-resolution 31P NMR) and outer membrane lipid composition, and are compared to the bactericidal susceptibility. LPS mutants show a progressive loss of resistance to killing by magainin 2 as the length of the LPS polysaccharide moiety decreases. Disordering of the outer membrane lipid fatty acyl chains by magainin 2, however, depends primarily upon the magnitude of PLS charge rather than the length of the LPS polysaccharide. While disruption of outer membrane structure most likely is not the primary factor leading to cell death, the susceptibility of Gram-negative cells to magainin 2 is associated with factors that facilitate the transport of the peptide across the outer membrane, such as the magnitude and location of LPS charge, and concentration of LPS in the outer membrane, outer membrane molecular architecture, and the presence or absence of the O-antigen side chainchain

  13. The lipopolysaccharide-activated innate immune response network of the horseshoe crab

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    S Kawabata

    2009-06-01

    Full Text Available Primary stimulation of the horseshoe crab innate immune system by bacterial lipopolysaccharide (LPS activates a network of responses to ensure host defense against invading pathogens. Granular hemocytes selectively respond to LPS via a G protein-dependent exocytic pathway that critically depends on the proteolytic activity of the LPS-responsive coagulation factor C. In response to stimulation by LPS, the hemocyte secretes transglutaminase (TGase and several kinds of defense molecules, such as coagulation factors, lectins, antimicrobial peptides, and protein substrates for TGase. LPS-induced hemocyte exocytosis is enhanced by a feedback mechanism in which the antimicrobial peptide tachyplesin serves as an endogenous mediator. The coagulation cascade triggered by LPS or ?-1,3-D-glucans results in the formation of coagulin fibrils that are subsequently stabilized by TGase-dependent cross-linking. A cuticle-derived chitin-binding protein additionally forms a TGase-stabilized mesh at sites of injury. Invading pathogens are agglutinated by both hemocyte- and plasma-derived lectins. In addition, the proclotting enzyme and tachyplesin functionally convert hemocyanin to phenoloxidase. In the plasma, coagulation factor C acts an LPS-sensitive complement C3 convertase on the surface of Gram-negative bacteria. In this manner, LPS-induced hemocyte exocytosis leads not only to coagulation but also activates a sophisticated innate immune response network that coordinately effects pathogen recognition, prophenoloxidase activation, pathogen clearance, and TGase-dependent wound healing

  14. Bordetella parapertussis PagP mediates the addition of two palmitates to the lipopolysaccharide lipid A.

    Science.gov (United States)

    Hittle, L E; Jones, J W; Hajjar, A M; Ernst, R K; Preston, A

    2015-02-01

    Bordetella bronchiseptica PagP (PagPBB) is a lipid A palmitoyl transferase that is required for resistance to antibody-dependent complement-mediated killing in a murine model of infection. B. parapertussis contains a putative pagP homolog (encoding B. parapertussis PagP [PagPBPa]), but its role in the biosynthesis of lipid A, the membrane anchor of lipopolysaccharide (LPS), has not been investigated. Mass spectrometry analysis revealed that wild-type B. parapertussis lipid A consists of a heterogeneous mixture of lipid A structures, with penta- and hexa-acylated structures containing one and two palmitates, respectively. Through mutational analysis, we demonstrate that PagPBPa is required for the modification of lipid A with palmitate. While PagPBB transfers a single palmitate to the lipid A C-3' position, PagPBPa transfers palmitates to the lipid A C-2 and C-3' positions. The addition of two palmitate acyl chains is unique to B. parapertussis. Mutation of pagPBPa resulted in a mutant strain with increased sensitivity to antimicrobial peptide killing and decreased endotoxicity, as evidenced by reduced proinflammatory responses via Toll-like receptor 4 (TLR4) to the hypoacylated LPS. Therefore, PagP-mediated modification of lipid A regulates outer membrane function and may be a means to modify interactions between the bacterium and its human host during infection. PMID:25422302

  15. The structure of the polysaccharide of the lipopolysaccharide produced by Taylorella equigenitalis type strain (ATCC 35865).

    Science.gov (United States)

    Vinogradov, Evgeny; MacLean, Leann L; Brooks, Brian W; Lutze-Wallace, Cheryl; Perry, Malcolm B

    2008-12-01

    Taylorella equigenitalis is a Gram-negative bacterium that causes venereally transmitted contagious equine metritis (CEM), and its identification and differentiation from other bacteria and Taylorella species is an important requirement for the control of CEM infection. Based on the results of NMR and MS analysis, the antigenic O-polysaccharide (O-PS) component of the lipopolysaccharide (LPS) produced by the type strain T. equigenitalis (ATCC 35865) was found to be a linear polymer composed of a repeating disaccharide unit, containing partially amidated 2,3-diacetamido-2,3-dideoxy-alpha-L-guluronic and 2,3-diacetamido-2,3-dideoxy-beta-D-mannuronic acids, terminated with a 4-O-methylated non-reducing Gulp-NAc3NAcA residue, and has the structure [structure: see text]. The O-PS of the type strain T. equigenitalis LPS provides a specific antigenic marker for the discrimination of the pathogen from the related type strain of T. asinigenitalis sp. nov, a phenotypically indistinguishable non-pathogenic bacterium having a serologically and structurally unrelated LPS O-antigen. The analysis of a structurally unusual core oligosaccharide of the LPS is also reported. PMID:18950750

  16. Structural analysis of the O-specific polysaccharide isolated from Plesiomonas shigelloides O51 lipopolysaccharide.

    Science.gov (United States)

    Maciejewska, Anna; Lukasiewicz, Jolanta; Niedziela, Tomasz; Szewczuk, Zbigniew; Lugowski, Czeslaw

    2009-05-12

    Plesiomonasshigelloides strain CNCTC 110/92 (O51) was identified as a new example of plesiomonads synthesising lipopolysaccharides (LPSs) that show preference for a non-aqueous surrounding during phenol/water extraction. Chemical analyses combined with (1)H and (13)C NMR spectroscopy, MALDI-TOF and ESI mass spectrometry showed that the repeating units of the O-specific polysaccharides isolated from phenol and water phase LPSs of P. shigelloides O51 have the same structure: -->4)-beta-D-GlcpNAc3NRA-(1-->4)-alpha-L-FucpAm3OAc-(1-->3)-alpha-D-QuipNAc-(1-->, containing the rare sugar constituent 2,3-diamino-2,3-dideoxyglucuronic acid (GlcpNAc3NRA), and substituents such as D-3-hydroxybutyric acid (R) and acetamidino group (Am). The HR-MAS NMR spectra obtained for the isolated LPSs and directly on bacteria indicated that the O-acetylation pattern was consistent throughout the entire preparation. The (1)H chemical shift values of the structure reporter groups identified in the isolated O-antigens matched those present in bacteria. We have found that the O-antigens recovered from the phenol phase showed a higher degree of polymerisation than those isolated from the water phase. PMID:19338978

  17. Cigarette smoke and lipopolysaccharide induce a proliferative airway smooth muscle phenotype

    Directory of Open Access Journals (Sweden)

    Zaagsma Johan

    2010-04-01

    Full Text Available Abstract Background A major feature of chronic obstructive pulmonary disease (COPD is airway remodelling, which includes an increased airway smooth muscle (ASM mass. The mechanisms underlying ASM remodelling in COPD are currently unknown. We hypothesized that cigarette smoke (CS and/or lipopolysaccharide (LPS, a major constituent of CS, organic dust and gram-negative bacteria, that may be involved in recurrent airway infections and exacerbations in COPD patients, would induce phenotype changes of ASM. Methods To this aim, using cultured bovine tracheal smooth muscle (BTSM cells and tissue, we investigated the direct effects of CS extract (CSE and LPS on ASM proliferation and contractility. Results Both CSE and LPS induced a profound and concentration-dependent increase in DNA synthesis in BTSM cells. CSE and LPS also induced a significant increase in BTSM cell number, which was associated with increased cyclin D1 expression and dependent on activation of ERK 1/2 and p38 MAP kinase. Consistent with a shift to a more proliferative phenotype, prolonged treatment of BTSM strips with CSE or LPS significantly decreased maximal methacholine- and KCl-induced contraction. Conclusions Direct exposure of ASM to CSE or LPS causes the induction of a proliferative, hypocontractile ASM phenotype, which may be involved in airway remodelling in COPD.

  18. Functional Identification of Proteus mirabilis eptC Gene Encoding a Core Lipopolysaccharide Phosphoethanolamine Transferase

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    Eleonora Aquilini

    2014-04-01

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

  19. Sympathetic glial cells and macrophages develop different responses to Trypanosoma cruzi infection or lipopolysaccharide stimulation

    Scientific Electronic Library Online (English)

    Camila Megale de, Almeida-Leite; Isabel Cristina Costa, Silva; Lúcia Maria da Cunha, Galvão; Rosa Maria Esteves, Arantes.

    2014-07-01

    Full Text Available Nitric oxide (NO) participates in neuronal lesions in the digestive form of Chagas disease and the proximity of parasitised glial cells and neurons in damaged myenteric ganglia is a frequent finding. Glial cells have crucial roles in many neuropathological situations and are potential sources of NO. [...] Here, we investigate peripheral glial cell response to Trypanosoma cruzi infection to clarify the role of these cells in the neuronal lesion pathogenesis of Chagas disease. We used primary glial cell cultures from superior cervical ganglion to investigate cell activation and NO production after T. cruzi infection or lipopolysaccharide (LPS) exposure in comparison to peritoneal macrophages. T. cruzi infection was greater in glial cells, despite similar levels of NO production in both cell types. Glial cells responded similarly to T. cruzi and LPS, but were less responsive to LPS than macrophages were. Our observations contribute to the understanding of Chagas disease pathogenesis, as based on the high susceptibility of autonomic glial cells to T. cruzi infection with subsequent NO production. Moreover, our findings will facilitate future research into the immune responses and activation mechanisms of peripheral glial cells, which are important for understanding the paradoxical responses of this cell type in neuronal lesions and neuroprotection.

  20. Lipopolysaccharide contamination of beta-lactoglobulin affects the immune response against intraperitoneally and orally administrated antigen

    DEFF Research Database (Denmark)

    Pedersen, Susanne Brix; Kjær, T.M.R.

    2004-01-01

    Microbial components in the environment are potent activators of the immune system with capacity to shift the active immune response towards priming of Th1 and/or Th2 cells. Lipopolysaccharide (LPS), a cell-wall component of Gram-negative bacteria, is extensively present in food products like cow's milk. It is not well established, however, how this presence of LPS affects oral tolerance induction. METHODS: We studied the effect of LPS contamination in a commercial preparation of the cow milk protein beta-lactoglobulin (beta-LG) on antigen-specific immune responses. IgG1/IgG2a production upon intraperitoneal immunization without adjuvant was measured, and oral tolerance induction against beta-LG after administration of either an aqueous solution or water-in-oil (w/o) emulsion of beta-LG was evaluated. RESULTS: LPS contamination of beta-LG provoked a beta-LG-specific IgG2a response, as well as an enhanced beta-LG-specific IgG1 response upon intraperitoneal immunization. Oral tolerance induction to beta-LG was induced by aqueous solutions of beta-LG with and without LPS administration. Conversely, oral administration of w/o-emulsified beta-LG prevented oral tolerance to beta-LG only when the beta-LG was contaminated with LPS. CONCLUSIONS: LPS contamination of an aqueous protein solution does not affect oral tolerance induction, whereas LPS present in emulsion prevents oral tolerance induction towards the food protein.

  1. Lipopolysaccharide contamination of beta-lactoglobulin affects the immune response against intraperitoneally and orally administered antigen

    DEFF Research Database (Denmark)

    Pedersen, Susanne Brix; Kjær, T.M.R.

    2004-01-01

    Microbial components in the environment are potent activators of the immune system with capacity to shift the active immune response towards priming of Th1 and/or Th2 cells. Lipopolysaccharide (LPS), a cell-wall component of Gram- negative bacteria, is extensively present in food products like cow's milk. It is not well established, however, how this presence of LPS affects oral tolerance induction. Methods: We studied the effect of LPS contamination in a commercial preparation of the cow milk protein beta-lactoglobulin (beta-LG) on antigen-specific immune responses. IgG1/IgG2a production upon intraperitoneal immunization without adjuvant was measured, and oral tolerance induction against beta-LG after administration of either an aqueous solution or water-in-oil (w/o) emulsion of beta-LG was evaluated. Results: LPS contamination of beta-LG provoked a beta-LG-specific IgG2a lresponse, as well as an enhanced beta-LG-specific IgG1 response upon intraperitoneal immunization. Oral tolerance induction to beta-LG was induced by aqueous solutions of beta-LG with and without LPS administration. Conversely, oral administration of w/o-emulsified beta-LG prevented oral tolerance to beta-LG only when the beta-LG was contaminated with LPS. Conclusions: LPS contamination of an aqueous protein solution does not affect oral tolerance induction, whereas LPS present in emulsion prevents oral tolerance induction towards the food protein.

  2. Effects of lipopolysaccharide-induced inflammation on expression of growth-associated genes by corticospinal neurons

    Directory of Open Access Journals (Sweden)

    Lieberman AR

    2006-01-01

    Full Text Available Abstract Background Inflammation around cell bodies of primary sensory neurons and retinal ganglion cells enhances expression of neuronal growth-associated genes and stimulates axonal regeneration. We have asked if inflammation would have similar effects on corticospinal neurons, which normally show little response to spinal cord injury. Lipopolysaccharide (LPS was applied onto the pial surface of the motor cortex of adult rats with or without concomitant injury of the corticospinal tract at C4. Inflammation around corticospinal tract cell bodies in the motor cortex was assessed by immunohistochemistry for OX42 (a microglia and macrophage marker. Expression of growth-associated genes c-jun, ATF3, SCG10 and GAP-43 was investigated by immunohistochemistry or in situ hybridisation. Results Application of LPS induced a gradient of inflammation through the full depth of the motor cortex and promoted c-Jun and SCG10 expression for up to 2 weeks, and GAP-43 upregulation for 3 days by many corticospinal neurons, but had very limited effects on neuronal ATF3 expression. However, many glial cells in the subcortical white matter upregulated ATF3. LPS did not promote sprouting of anterogradely labelled corticospinal axons, which did not grow into or beyond a cervical lesion site. Conclusion Inflammation produced by topical application of LPS promoted increased expression of some growth-associated genes in the cell bodies of corticospinal neurons, but was insufficient to promote regeneration of the corticospinal tract.

  3. Chronic lipopolysaccharide infusion fails to induce depressive-like behaviour in adult male rats

    DEFF Research Database (Denmark)

    Fischer, Christina Weide; Liebenberg, Nico

    2015-01-01

    BACKGROUND: Chronic inflammation is implicated in numerous diseases, including major depression and type 2 diabetes mellitus (T2DM). Since depression and T2DM often co-exist, inflammatory pathways are suggested as a possible link. Hence, the establishment of an immune-mediated animal model would shed light on mechanisms possibly linking depression and metabolic alterations. OBJECTIVE: In this study we investigated a behavioural and metabolic paradigm following chronic infusion with low doses of lipopolysaccharide (LPS) using osmotic minipumps in male rats. METHODS: Behavioural testing consisted of evaluating activity level in the open field and depressive-like behaviour in the forced swim test. Metabolic assessment included measurement of body weight, food and water intake, and glucose and insulin levels during an oral glucose tolerance test. RESULTS: LPS-infused rats showed acute signs of sickness behaviour, but chronic LPS infusion did not induce behavioural or metabolic changes. CONCLUSION: These results suggest that although inflammation is immediately induced as indicated by acute sickness, 4 weeks of chronic LPS administration via osmotic minipumps did not result in behavioural changes. Therefore, this paradigm may not be a suitable model for studying the underlying mechanisms that link depression and T2DM.

  4. Metabonomic evaluation of idiosyncrasy-like liver injury in rats cotreated with ranitidine and lipopolysaccharide

    International Nuclear Information System (INIS)

    Idiosyncratic liver injury occurs in a small fraction of people on certain drug regimens. The cause of idiosyncratic hepatotoxicity is not known; however, it has been proposed that environmental factors such as concurrent inflammation initiated by bacterial lipopolysaccharide (LPS) increase an individual's susceptibility to drug toxicity. Ranitidine (RAN), a histamine-2 receptor antagonist, causes idiosyncratic liver injury in humans. In a previous report, idiosyncrasy-like liver toxicity was created in rats by cotreating them with LPS and RAN. In the present study, the ability of metabonomic techniques to distinguish animals cotreated with LPS and RAN from those treated with each agent individually was investigated. Rats were treated with LPS or its vehicle and with RAN or its vehicle, and urine was collected for nuclear magnetic resonance (NMR)- and mass spectroscopy-based metabonomic analyses. Blood and liver samples were also collected to compare metabonomic results with clinical chemistry and histopathology. NMR metabonomic analysis indicated changes in the pattern of metabolites consistent with liver damage that occurred only in the LPS/RAN cotreated group. Principal component analysis of urine spectra by either NMR or mass spectroscopy produced a clear separation of the rats treated with LPS/RAN from the other three groups. Clinical chemistry (serum alanine aminotransferase and aspartate aminotransferase activities) and histopathology corroborated these results histopathology corroborated these results. These findings support the potential use of a noninvasive metabonomic approach to identify drug candidates with potential to cause idiosyncratic liver toxicity with inflammagen coexposure

  5. Mechanism of attenuation by beta-hydroxy-beta-methylbutyrate of muscle protein degradation induced by lipopolysaccharide.

    Science.gov (United States)

    Russell, Steven T; Tisdale, Michael J

    2009-10-01

    The mechanism of the effect of beta-hydroxy-beta-methylbutyrate (HMB) on protein degradation induced by lipopolysaccharide (LPS) has been evaluated in murine myotubes. HMB (50 muM) completely attenuated total protein degradation induced by LPS (1-100 ng/ml), formation of reactive oxygen species (ROS) and activation of caspase-3/-8. Specific inhibitors of caspase-3/-8 completely attenuated ROS production, total protein degradation and the LPS-induced autophosphorylation of dsRNA-dependent protein kinase (PKR). Protein degradation in response to LPS or ROS production was not seen in myotubes transfected with mutant PKRDelta6, suggesting that PKR was involved in ROS production, which was essential for total protein degradation. This was confirmed using the antioxidant butylated hydroxytoluene (BHT) which completely attenuated protein degradation in response to LPS. The link between PKR activation and ROS production was mediated through p38 mitogen-activated protein kinase (MAPK), which was activated by LPS in myotubes transfected with wild-type PKR, but not PKRDelta6. Both ROS production and protein degradation induced by LPS were completely attenuated by SB203580, a specific inhibitor of p38MAPK. This suggests that LPS induces protein degradation through a signalling cascade involving activation of caspase-3/-8, activation of PKR and production of ROS through p38MAPK, and that this process is attenuated by HMB. PMID:19404720

  6. Static magnetic field attenuates lipopolysaccharide-induced inflammation in pulp cells by affecting cell membrane stability.

    Science.gov (United States)

    Hsieh, Sung-Chih; Tsao, Jeng-Ting; Lew, Wei-Zhen; Chan, Ya-Hui; Lee, Lin-Wen; Lin, Che-Tong; Huang, Yung-Kai; Huang, Haw-Ming

    2015-01-01

    One of the causes of dental pulpitis is lipopolysaccharide- (LPS-) induced inflammatory response. Following pulp tissue inflammation, odontoblasts, dental pulp cells (DPCs), and dental pulp stem cells (DPSCs) will activate and repair damaged tissue to maintain homeostasis. However, when LPS infection is too serious, dental repair is impossible and disease may progress to irreversible pulpitis. Therefore, the aim of this study was to examine whether static magnetic field (SMF) can attenuate inflammatory response of dental pulp cells challenged with LPS. In methodology, dental pulp cells were isolated from extracted teeth. The population of DPSCs in the cultured DPCs was identified by phenotypes and multilineage differentiation. The effects of 0.4?T SMF on DPCs were observed through MTT assay and fluorescent anisotropy assay. Our results showed that the SMF exposure had no effect on surface markers or multilineage differentiation capability. However, SMF exposure increases cell viability by 15%. In addition, SMF increased cell membrane rigidity which is directly related to higher fluorescent anisotropy. In the LPS-challenged condition, DPCs treated with SMF demonstrated a higher tolerance to LPS-induced inflammatory response when compared to untreated controls. According to these results, we suggest that 0.4?T SMF attenuates LPS-induced inflammatory response to DPCs by changing cell membrane stability. PMID:25884030

  7. Lipopolysaccharide stimulation improves the odontoblastic differentiation of human dental pulp cells.

    Science.gov (United States)

    Huang, Yihua; Jiang, Hongwei; Gong, Qimei; Li, Xuyan; Ling, Junqi

    2015-05-01

    Lipopolysaccharide (LPS) is one of the causative agents of pulpitis and previous studies have demonstrated that the LPS stimulation of human aortic valve interstitial cells induces inflammatory mediators and the gene expression of osteogenic factors. Therefore, in the present study, it was hypothesized that LPS affects the odontoblastic differentiation of human dental pulp cells (hDPCs). In order to investigate this, an in vitro study using hDPCs was performed. Increased alkaline phosphatase (ALP) activity was observed in the hDPCs treated with LPS, which was more marked when the cells were costimulated with odontogenic induction medium (OM). LPS also appeared to increase the gene expression levels of dentin sialophosphoprotein and dentin matrix protein?1 and the protein expression level of dental sialoprotein in the hDPCs, particularly in combination with OM. In addition, the size and the number of nodules formed in the hDPCs exposed to OM and LPS were increased compared to those stimulated by OM alone. To determine the role of nuclear factor ?B (NF??B) during the LPS?induced odontoblastic differentiation of hDPCs, immunofluorescence was performed. The nuclear translocation of NF??B, induced by LPS was confirmed, suggesting its involvement in the LPS?induced increase in odontoblastic differentiation of hDPCs. In conclusion, there may be an association between LPS stimulation, with or without OM, and odontoblastic differentiation. PMID:25528991

  8. Static Magnetic Field Attenuates Lipopolysaccharide-Induced Inflammation in Pulp Cells by Affecting Cell Membrane Stability

    Science.gov (United States)

    Tsao, Jeng-Ting; Lee, Lin-Wen; Lin, Che-Tong

    2015-01-01

    One of the causes of dental pulpitis is lipopolysaccharide- (LPS-) induced inflammatory response. Following pulp tissue inflammation, odontoblasts, dental pulp cells (DPCs), and dental pulp stem cells (DPSCs) will activate and repair damaged tissue to maintain homeostasis. However, when LPS infection is too serious, dental repair is impossible and disease may progress to irreversible pulpitis. Therefore, the aim of this study was to examine whether static magnetic field (SMF) can attenuate inflammatory response of dental pulp cells challenged with LPS. In methodology, dental pulp cells were isolated from extracted teeth. The population of DPSCs in the cultured DPCs was identified by phenotypes and multilineage differentiation. The effects of 0.4?T SMF on DPCs were observed through MTT assay and fluorescent anisotropy assay. Our results showed that the SMF exposure had no effect on surface markers or multilineage differentiation capability. However, SMF exposure increases cell viability by 15%. In addition, SMF increased cell membrane rigidity which is directly related to higher fluorescent anisotropy. In the LPS-challenged condition, DPCs treated with SMF demonstrated a higher tolerance to LPS-induced inflammatory response when compared to untreated controls. According to these results, we suggest that 0.4?T SMF attenuates LPS-induced inflammatory response to DPCs by changing cell membrane stability. PMID:25884030

  9. Production and characterization of monoclonal antibodies against the O-5 antigen of Salmonella typhimurium lipopolysaccharide.

    Science.gov (United States)

    Jaradat, Z W; Zawistowski, J

    1996-01-01

    Three murine monoclonal antibodies (MAbs) were produced by fusion of P3X63-Ag8.653 myeloma cells and splenocytes of a mouse immunized with heat-attenuated (20 min, 80 degrees C) Salmonella typhimurium cells. MAbs 5A5 and 5B2 were of the immunoglobulin M (IgM) class, while MAb 4A8 was IgG2a. All possessed the kappa light chains. The MAbs were specific to the lipopolysaccharide (LPS) O-5 antigen of Salmonella B serogroup, as determined by electrophoresis followed by immunoblotting. All MAbs recognized the same epitope, as determined by an additive enzyme-linked immunosorbent assay (ELISA), although IgM MAbs exhibited higher avidity than the IgG MAb. ELISA analyses revealed that all three MAbs reacted with S. typhimurium (LPS O:1, 4, 5, and 12) while failing to recognize S. typhimurium var. copenhagen (LPS O:1, 4, and 12). The MAbs reacted equally with live and heat-attenuated Salmonella B serovars containing LPS O-5 antigen. The ability of the MAbs to detect live bacterial cells was further confirmed by transmission electron microscopy. Treatment of bacteria with cholic acid and extremely low pH did not affect antibody binding to S. typhimurium. However, when S. typhimurium cells were exposed to alkaline conditions prior to reaction with all three MAbs, no binding was observed. The use of MAbs to discriminate between S. typhimurium and S. typhimurium var. copenhagen in meat samples was investigated. PMID:8572685

  10. Atorvastatin reduces lipopolysaccharide-induced expression of cyclooxygenase-2 in human pulmonary epithelial cells

    Directory of Open Access Journals (Sweden)

    Chen Ping

    2005-04-01

    Full Text Available Abstract Objective To explore the effects of atorvastatin on expression of cyclooxygenase-2 (COX-2 in human pulmonary epithelial cells (A549. Methods A549 cells were incubated in DMEM medium containing lipopolysaccharide (LPS in the presence or absence of atorvastatin. After incubation, the medium was collected and the amount of prostaglandin E2 (PGE2 was measured by enzyme-linked immunosorbent assay (ELISA. The cells were harvested, and COX-2 mRNA and protein were analyzed by RT-PCR and western-blot respectively. Results LPS increased the expression of COX-2 mRNA and production of PGE2 in a dose- and time-dependent manner in A549. Induction of COX-2 mRNA and protein by LPS were inhibited by atorvastatin in a dose-dependent manner. Atorvastatin also significantly decreased LPS-induced production of PGE2. There was a positive correlation between reduced of COX-2 mRNA and decreased of PGE2 (r = 0.947, P Conclusion Atorvastatin down-regulates LPS-induced expression of the COX-2 and consequently inhibits production of PGE2 in cultured A549 cells.

  11. Lipopolysaccharide-induced multinuclear cells: Increased internalization of polystyrene beads and possible signals for cell fusion

    Energy Technology Data Exchange (ETDEWEB)

    Nakanishi-Matsui, Mayumi, E-mail: nakanim@iwate-med.ac.jp; Yano, Shio; Futai, Masamitsu

    2013-11-01

    Highlights: •LPS induces multinuclear cells from murine macrophage-derived RAW264.7 cells. •Large beads are internalized by cells actively fusing to become multinuclear. •The multinuclear cell formation is inhibited by anti-inflammatory cytokine, IL10. •Signal transduction for cell fusion is different from that for inflammation. -- Abstract: A murine macrophage-derived line, RAW264.7, becomes multinuclear on stimulation with lipopolysaccharide (LPS), an outer membrane component of Gram-negative bacteria. These multinuclear cells internalized more polystyrene beads than mononuclear cells or osteoclasts (Nakanishi-Matsui, M., Yano, S., Matsumoto, N., and Futai, M., 2012). In this study, we analyzed the time courses of cell fusion in the presence of large beads. They were internalized into cells actively fusing to become multinuclear. However, the multinuclear cells once formed showed only low phagocytosis activity. These results suggest that formation of the multinuclear cells and bead internalization took place simultaneously. The formation of multinuclear cells was blocked by inhibitors for phosphoinositide 3-kinase, phospholipase C, calcineurin, and c-Jun N-terminal kinase. In addition, interleukin 6 and 10 also exhibited inhibitory effects. These signaling molecules and cytokines may play a crucial role in the LPS-induced multinuclear cell formation.

  12. The anti-inflammatory effects of methylsulfonylmethane on lipopolysaccharide-induced inflammatory responses in murine macrophages.

    Science.gov (United States)

    Kim, Yoon Hee; Kim, Dae Hwan; Lim, Hwan; Baek, Doo-Yeon; Shin, Hyun-Kyung; Kim, Jin-Kyung

    2009-04-01

    Methylsulfonylmethane (MSM), also known as dimethyl sulfone and methyl sulfone, is an organic sulfur-containing compound that occurs naturally in a variety of fruits, vegetables, grains, and animals, including humans. In the present study, we demonstrated the anti-inflammatory effects of MSM in lipopolysaccharide (LPS)-stimulated murine macrophages, RAW264.7 cells. MSM significantly inhibited the release of nitric oxide and prostaglandin E(2) by alleviating the expression of inducible nitric oxide synthase and cyclooxygenase-2 in LPS-stimulated RAW264.7 cells. Furthermore, the levels of interleukin-6 and tumor necrosis factor-alpha were decreased by MSM treatment in cell culture supernatants. Further study indicated that the translocation of the p65 subunit of nuclear factor (NF)-kappaB to the nucleus was inhibited by MSM treatment in LPS-stimulated RAW264.7 cells, in which it helped block degradation of inhibitor of NF-kappaB. In addition, in vivo studies demonstrated that topical administration of MSM at 500-1250 microg/ear resulted in similar inhibitory activities in 12-O-tetradecanoylphorbol 13-acetate-induced mouse ear edema. Collectively, theses results indicate that MSM inhibits LPS-induced release of pro-inflammatory mediators in murine macrophages through downregulation of NF-kappaB signaling. PMID:19336900

  13. AKR1B10 is induced by hyperglycaemia and lipopolysaccharide in patients with diabetic nephropathy.

    Science.gov (United States)

    Shaw, Nicholas; Yang, Bingmei; Millward, Ann; Demaine, Andrew; Hodgkinson, Andrea

    2014-03-01

    Aldose reductase family member B10 (AKR1B10) belongs to the aldo-keto reductase gene superfamily and is closely related to aldose reductase (AKR1B1). It has been shown that AKR1B10 is present in many of the same human tissues as AKR1B1. The objective of this study was to investigate whether AKR1B10 has a role in diabetic nephropathy (DN) by investigating its response to high glucose and inflammation, both of which have been associated with the development and progression of DN. Expression levels of AKR1B10 were determined in peripheral blood mononuclear cells (PBMCs) obtained from 25 patients with type 1 diabetes and nephropathy, 25 without DN and 25 normal healthy controls that were exposed to high glucose (25 mM D-glucose) and also the inflammatory stressor lipopolysaccharide (LPS, 10 ?m). Under high glucose and LPS conditions, there was a significant increase in the expression of AKR1B10 in the PBMCs from patients with DN compared to those without DN and the normal controls. In conclusion, these results suggest that AKR1B10 may have an important role in the development and progression of DN. PMID:23975544

  14. Protective effect of magnolol on lipopolysaccharide-induced acute lung injury in mice.

    Science.gov (United States)

    Ni, Yun Feng; Jiang, Tao; Cheng, Qing Shu; Gu, Zhong Ping; Zhu, Yi Fang; Zhang, Zhi Pei; Wang, Jian; Yan, Xiao Long; Wang, Wu Ping; Ke, Chang Kang; Han, Yong; Li, Xiao Fei

    2012-12-01

    Magnolol, a tradition Chinese herb, displays an array of activities including antifungal, antibacterial, and antioxidant effects. To investigate the protective effect of magnolol on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice. ALI was induced in mice by intratracheal instillation of LPS (1 mg/kg). The mice received intratracheal instillation of magnolol (5 ?g/kg) 30 min before LPS administration. Pulmonary histological changes were evaluated by hematoxylin-eosin stain and lung wet/dry weight ratios were observed. Concentrations of tumor necrosis factor (TNF)-? and interleukin (IL)-1?, and myeloperoxidase (MPO) activity were measured by enzyme-linked immunosorbent assay. Expression of cyclooxygenase (COX)-2 in lung tissues was determined by Western blot analysis. Magnolol pretreatment significantly attenuated the severity of lung injury and inhibited the production of TNF-? and IL-1? in mice with ALI. After LPS administration, the lung wet/dry weight ratios, as an index of lung edema, and MPO activity were also markedly reduced by magnolol pretreatment. The expression of COX-2 was significantly suppressed by magnolol pretreatment. Magnolol potently protected against LPS-induced ALI and the protective effects of magnolol may attribute partly to the suppression of COX-2 expression. PMID:23053725

  15. Antimicrobial Peptides: Insights into Membrane Permeabilization, Lipopolysaccharide Fragmentation and Application in Plant Disease Control.

    Science.gov (United States)

    Datta, Aritreyee; Ghosh, Anirban; Airoldi, Cristina; Sperandeo, Paola; Mroue, Kamal H; Jiménez-Barbero, Jesús; Kundu, Pallob; Ramamoorthy, Ayyalusamy; Bhunia, Anirban

    2015-01-01

    The recent increase in multidrug resistance against bacterial infections has become a major concern to human health and global food security. Synthetic antimicrobial peptides (AMPs) have recently received substantial attention as potential alternatives to conventional antibiotics because of their potent broad-spectrum antimicrobial activity. These peptides have also been implicated in plant disease control for replacing conventional treatment methods that are polluting and hazardous to the environment and to human health. Here, we report de novo design and antimicrobial studies of VG16, a 16-residue active fragment of Dengue virus fusion peptide. Our results reveal that VG16KRKP, a non-toxic and non-hemolytic analogue of VG16, shows significant antimicrobial activity against Gram-negative E. coli and plant pathogens X. oryzae and X. campestris, as well as against human fungal pathogens C. albicans and C. grubii. VG16KRKP is also capable of inhibiting bacterial disease progression in plants. The solution-NMR structure of VG16KRKP in lipopolysaccharide features a folded conformation with a centrally located turn-type structure stabilized by aromatic-aromatic packing interactions with extended N- and C-termini. The de novo design of VG16KRKP provides valuable insights into the development of more potent antibacterial and antiendotoxic peptides for the treatment of human and plant infections. PMID:26144972

  16. The effect of bacterial lipopolysaccharide on gastric emptying in rats suffering from moderate renal insufficiency

    Scientific Electronic Library Online (English)

    S.Z.P., Rigatto; E.F., Collares.

    1998-04-01

    Full Text Available The objective of the present study was to evaluate the response of rats suffering from moderate renal insufficiency to bacterial lipopolysaccharide (LPS, or endotoxin). The study involved 48 eight-week-old male SPF Wistar rats (175-220 g) divided into two groups of 24 animals each. One group underwe [...] nt 5/6 nephrectomy while the other was sham-operated. Two weeks after surgery, the animals were further divided into two subgroups of 12 animals each and were fasted for 20 h but with access to water ad libitum. One nephrectomized and one sham-treated subgroup received E. coli LPS (25 µg/kg, iv) while the other received a sterile, pyrogen-free saline solution. Gastric retention (GR) was determined 10 min after the orogastric infusion of a standard saline test meal labeled with phenol red (6 mg/dl). The gastric emptying of the saline test meal was studied after 2 h. Renal function was evaluated by measuring the plasma levels of urea and creatinine. The levels of urea and creatinine in 5/6 nephrectomized animals were two-fold higher than those observed in the sham-operated rats. Although renal insufficiency did not change gastric emptying (median %GR = 26.6 for the nephrectomized subgroup and 29.3 for the sham subgroup), LPS significantly retarded the gastric emptying of the sham and nephretomized groups (median %GR = 42.0 and 61.0, respectively), and was significantly greater (P

  17. The roots of Nardostachys jatamansi inhibits lipopolysaccharide-induced endotoxin shock.

    Science.gov (United States)

    Bae, Gi-Sang; Seo, Sang-Wan; Kim, Min-Sun; Park, Kyoung-Chel; Koo, Bon Soon; Jung, Won-Seok; Cho, Gil-Hwan; Oh, Hyun Cheol; Yun, Seung-Won; Kim, Jong-Jin; Kim, Sung Gyu; Hwang, Sung-Yeon; Song, Ho-Joon; Park, Sung-Joo

    2011-01-01

    Nardostachys jatamansi (NJ) has been used in the treatment of inflammatory diseases. However, it is not clear how NJ produces anti-inflammatory effects. In the present study, using an experimental model of lipopolysaccharide (LPS)-induced endotoxin shock, the protective effects and mechanisms of action of NJ were investigated. The water extract of roots of NJ was administrated to mice orally (1, 5, and 10 mg/kg) 1 h after or before LPS challenge. The administration of NJ inhibited LPS-induced endotoxin shock and the production of inflammatory mediators, such as interleukin (IL)-1?, IL-6, tumor necrosis factor (TNF)-?, and interferon (IFN)-?/?. Murine peritoneal macrophages were used to determine the production of inflammatory mediators. In peritoneal macrophages, NJ also inhibited LPS-induced production of inflammatory mediators, such as IL-1?, IL-6, TNF-?, and IFN-?/?. In addition, NJ reduced the activation of mitogen-activated protein kinases (MAPKs) and the level of expression of interferon regulatory factor (IRF)-1 and IRF-7 mRNA. Furthermore, post-treatment with NJ reduced LPS-induced endotoxin shock and the production of inflammatory mediators. These results suggest that NJ inhibits endotoxin shock by inhibiting the production of IL-1?, IL-6, TNF-?, and IFN-?/? through the inhibition of MAPKs activation and IRF induction. PMID:20799070

  18. Beneficial Effects of Fractions of Nardostachys jatamansi on Lipopolysaccharide-Induced Inflammatory Response.

    Science.gov (United States)

    Bae, Gi-Sang; Heo, Kwang-Ho; Choi, Sun Bok; Jo, Il-Joo; Kim, Dong-Goo; Shin, Joon-Yeon; Seo, Seung-Hee; Park, Kyoung-Chel; Lee, Dong-Sung; Oh, Hyuncheol; Kim, Youn-Chul; Song, Ho-Joon; Shin, Byung-Cheul; Park, Sung-Joo

    2014-01-01

    It has been previously shown that Nardostachys jatamansi (NJ) exhibits anti-inflammatory properties against lipopolysaccharide (LPS) challenges. However, the potency of NJ constituents against LPS-induced inflammatory responses has not been examined. In this present study, we determined which NJ extract fractions exhibit inhibitory effects against LPS-induced inflammatory responses. Among the NJ fractions, NJ-1, NJ-3, NJ-4, and NJ-6 inhibited LPS-induced production of NO. The NJ-3, NJ-4, and NJ-6 fractions also inhibited the production of cytokines, such as IL-1 ? , IL-6, and TNF- ? . However, NJ-1, NJ-3, NJ-4, and NJ-6 showed differential inhibitory mechanisms against LPS-induced inflammatory responses. NJ-1, NJ-3, and NJ-4 inhibited LPS-induced activation of c-jun NH2-terminal kinase (JNK) and p38 but did not affect activation of extracellular signal-regulated kinase (ERK) or NF- ? B. On the other hand, NJ-6 inhibited activation of MAPKs and NF- ? B. In addition, in vivo experiments revealed that administration of NJ-1, NJ-3, NJ-4, and NJ-6 reduced LPS-induced endotoxin shock, with NJ-6 especially showing a marked protective effect. Taken together, these results provide the evidence for the potential of selective NJ fractions against LPS-induced inflammation. Thus, it will be advantageous to further isolate and determine single effective compounds from these potent fractions. PMID:24795771

  19. Antagonism by salvianolic acid B of lipopolysaccharide-induced disseminated intravascular coagulation in rabbits.

    Science.gov (United States)

    Wu, Zheng; Li, Jian-Nan; Bai, Zhi-Quan; Lin, Xi

    2014-07-01

    The aim of the present study was to investigate the effects of salvianolic acid B on lipopolysaccharide (LPS)-induced disseminated intravascular coagulation (DIC) in rabbits. Continuous infusion of LPS was used to induce a DIC model in rabbits. Treatment with salvianolic acid B (1, 3 or 6 mg/kg) was started simultaneously with LPS infusion (0.5 mg/kg LPS in 60 mL saline; 10 mL/h over a period of 6 h) through the contralateral marginal ear vein. Activated partial thromboplastin time (APTT), prothrombin time (PT), platelet count and fibrinogen concentration were determined, as were plasma levels of fibrin-fibrinogen degradation products (FDP), alanine aminotransferase (ALT), blood urea nitrogen (BUN), protein C activity, antithrombin III (ATIII) and tumour necrosis factor (TNF)-? concentration. The gradual impairment of haemostatic parameters was induced by continuous infusion of LPS. There were marked increases in APTT, PT, BUN, ALT and plasma TNF-? and marked decreases in the platelet count, fibrinogen, FDP, protein C and ATIII. The intravenous administration of 1, 3 or 6 mg/kg salvianolic acid B attenuated the increases in APTT, PT, BUN, ALT and plasma TNF-? and the decreases in fibrinogen, platelet, FDP, protein C and ATIII induced by LPS infusion. These observations indicate that salvianolic acid B has an effect against LPS-induced DIC in rabbits. PMID:24739088

  20. Regulation of HIV receptor expression in cervical epithelial cells by Gram-negative bacterial lipopolysaccharide

    Scientific Electronic Library Online (English)

    K J, Sales; T, Klein; A A, Katz.

    2015-01-01

    Full Text Available BACKGROUND: Sexually transmitted infections (STIs) caused by the Gram-negative bacteria Chlamydia trachomatis and Neisseria gonorrhoeae are associated with an increased risk of HIV acquisition in South African women. HIV infection involves binding of the virus to CD4+ receptors on host cells and sub [...] sequent binding to a chemokine co-receptor that mediates fusion with the host target cell membrane. OBJECTIVE: To investigate the potential impact of STIs on HIV receptor expression in cervical epithelial cells, and the molecular pathways mediating this effect. METHODS: Expression of Toll-like receptor 4 (TLR4), CD4+ and CCR5 was investigated in HPV type 18-positive (HeLa) and HPV-negative (C33A) cervical epithelial cells, uterine adenocarcinoma cells (Ishikawa), cervical squamous cell carcinoma tissue and normal cervical tissue by real-time polymerase chain reaction (RT-PCR) analysis. HIV receptor expression in HeLa cells was investigated in the presence/absence of 10 ?g/mL bacterial lipopolysaccharide (LPS) and chemical inhibitors of epidermal growth factor receptor (EGFR), extracellular signal-regulated kinase (ERK1/2) or cyclo-oxygenase-2 (COX-2) by RT-PCR analysis. RESULTS: TLR4, CD4+ and CCR5 expression was elevated in HeLa, C33A and Ishikawa cell lines and carcinoma tissue, compared with normal cervical tissue. Treatment of HeLa cells with LPS increased expression of the primary HIV chemokine co-receptor CCR5 (p

  1. Hepatoprotective effect of cryptotanshinone from Salvia miltiorrhiza in D-galactosamine/lipopolysaccharide-induced fulminant hepatic failure.

    Science.gov (United States)

    Jin, Quan; Jiang, Shuang; Wu, Yan-Ling; Bai, Ting; Yang, Yong; Jin, Xuejun; Lian, Li-Hua; Nan, Ji-Xing

    2014-01-15

    Cryptotanshinone from Salvia miltiorrhiza Bunge was investigated for hepatoprotective effects in d-galactosamine (GalN)/lipopolysaccharide (LPS)-induced fulminant hepatic failure. Cryptotanshinone (20 or 40 mg/kg) was orally administered 12 and 1h prior to GalN (700 mg/kg)/LPS (10 ?g/kg) injection. The increased mortality and TNF-? levels by GalN/LPS were declined by cryptotanshinone pretreatment. In addition, cryptotanshinone attenuated GalN/LPS-induced apoptosis, characterized by the blockade of caspase-3, -8, and -9 activation, as well as the release of cytochrome c from the mitochondria. In addition, cryptotanshinone significantly suppressed JNK, ERK and p38 phosphorylation induced by GalN/LPS, and phosphorylation of TAK1 as well. Furthermore, cryptotanshinone significantly inhibited the activation of NF-?B and suppressed the production of proinflammatory cytokines. These findings suggested that hepatoprotective effect of cryptotanshinone is likely associated with its anti-apoptotic activity and the down-regulation of MAPKs and NF-?B associated at least in part with suppressing TAK1 phosphorylation. PMID:24011530

  2. Rough-Form-Lipopolysaccharide Increase Apoptosis in Human CD4(+) and CD8(+) T-Lymphocytes

    DEFF Research Database (Denmark)

    Nielsen, Jeppe Sylvest; Larsson, Anders

    2012-01-01

    Immunosuppression induced by lymphocyte apoptosis is considered an important factor in the pathogenesis of sepsis and has been demonstrated in both animal models of LPS-induced endotoxemia and septic patients. Since rough-form lipopolysaccharide (R-LPS) recently have been shown to elicit a stronger immunological response than regular smooth-form LPS (S-LPS), we aimed to assess the apoptosis-inducing capabilities of R-LPS in different subsets of lymphocytes (CD4(+) T-cells, CD8(+) T-cell, B-cells, and NK-cells). Using multicolor flow cytometry on human peripheral blood mononuclear cells, we found that R-LPS increased apoptosis in CD4(+) and CD8(+) T-cells assessed by annexin V and propidium iodide (AV(+) PI(-) ), compared to both S-LPS and unstimulated cells. 7-amino-actinomycin D (7-AAD) and staining for intracellular active caspase-3, which are considered later signs of apoptosis, did not reveal the same results. Both forms appeared to inhibit apoptosis in B-cells, but no LPS-form-specific effect was seen onB- or NK cells. Our results indicate that R-LPS induce a stronger AV(+) PI(-) assessed apoptotic response in T-cells than S-LPS. Our findings emphasize the importance of T-cell apoptosis in endotoxemia and advocates for control of LPS-form in both endotoxemia research and clinical trials with Gram-negative infections.

  3. Protective effect of linalool against lipopolysaccharide/D-galactosamine-induced liver injury in mice.

    Science.gov (United States)

    Li, Jingyuan; Zhang, Xiaoyu; Huang, Haiying

    2014-12-01

    Linalool, a natural compound of the essential oils, has been shown to have antinociceptive, antimicrobial, and anti-inflammatory properties. The aim of this study was to investigate the effects of linalool against lipopolysaccharide (LPS)/D-galactosamine (GalN)-induced liver injury in mice. Mice were administered with linalool 1h before receiving LPS (50 ?g/kg) and GalN (800 mg/kg). The results demonstrated that linalool had a protective effect on LPS/GalN-induced acute liver injury, as evidenced by the attenuation of hepatic pathological damage, malondialdehyde (MDA) content, MPO activity and serum ALT and AST levels. Linalool alleviated serum and hepatic TNF-? and IL-6 production, as well as hepatic iNOS and COX-2 expression by inhibiting NF-?B activation. Treatment of linalool increased bcl-2 expression and inhibited caspase-3 and caspase-8 expression. In addition, linalool increased Nrf2 and heme oxygenase-1 expression up-regulation by LPS/GalN. In conclusion, our results suggested that linalool was protected against LPS/GalN-induced liver injury through induction of antioxidant defense via Nrf2 activating and reduction inflammatory response via NF-?B inhibition. PMID:25311666

  4. Effects of a neuronal nitric oxide synthase inhibitor on lipopolysaccharide-induced fever

    Scientific Electronic Library Online (English)

    C.A.A., Perotti; M.S., Nogueira; J., Antunes-Rodrigues; E.C., Cárnio.

    1999-11-01

    Full Text Available It has been demonstrated that nitric oxide (NO) has a thermoregulatory action, but very little is known about the mechanisms involved. In the present study we determined the effect of neuronal nitric oxide synthase (nNOS) inhibition on thermoregulation. We used 7-nitroindazole (7-NI, 1, 10 and 30 mg [...] /kg body weight), a selective nNOS inhibitor, injected intraperitoneally into normothermic Wistar rats (200-250 g) and rats with fever induced by lipopolysaccharide (LPS) (100 µg/kg body weight) administration. It has been demonstrated that the effects of 30 mg/kg of 7-NI given intraperitoneally may inhibit 60% of nNOS activity in rats. In all experiments the colonic temperature of awake unrestrained rats was measured over a period of 5 h at 15-min intervals after intraperitoneal injection of 7-NI. We observed that the injection of 30 mg/kg of 7-NI induced a 1.5oC drop in body temperature, which was statistically significant 1 h after injection (P

  5. Effects of propofol on lipopolysaccharide-induced expression and release of HMGB1 in macrophages

    Scientific Electronic Library Online (English)

    T., Wang; X.Y., Wei; B., Liu; L.J., Wang; L.H., Jiang.

    2015-04-01

    Full Text Available This study aimed to determine the effects of different concentrations of propofol (2,6-diisopropylphenol) on lipopolysaccharide (LPS)-induced expression and release of high-mobility group box 1 protein (HMGB1) in mouse macrophages. Mouse macrophage cell line RAW264.7 cells were randomly divided into [...] 5 treatment groups. Expression levels of HMGB1 mRNA were detected using RT-PCR, and cell culture supernatant HMGB1 protein levels were detected using enzyme-linked immunosorbent assay (ELISA). Translocation of HMGB1 from the nucleus to the cytoplasm in macrophages was observed by Western blotting and activity of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-?B) in the nucleus was detected using ELISA. HMGB1 mRNA expression levels increased significantly in the cell culture supernatant and in cells after 24 h of stimulating RAW264.7 cells with LPS (500 ng/mL). However, HMGB1 mRNA expression levels in the P2 and P3 groups, which received 500 ng/mL LPS with 25 or 50 ?mol/mL propofol, respectively, were significantly lower than those in the group receiving LPS stimulation (P

  6. Effects of propofol on lipopolysaccharide-induced expression and release of HMGB1 in macrophages

    Scientific Electronic Library Online (English)

    T., Wang; X.Y., Wei; B., Liu; L.J., Wang; L.H., Jiang.

    Full Text Available This study aimed to determine the effects of different concentrations of propofol (2,6-diisopropylphenol) on lipopolysaccharide (LPS)-induced expression and release of high-mobility group box 1 protein (HMGB1) in mouse macrophages. Mouse macrophage cell line RAW264.7 cells were randomly divided into [...] 5 treatment groups. Expression levels of HMGB1 mRNA were detected using RT-PCR, and cell culture supernatant HMGB1 protein levels were detected using enzyme-linked immunosorbent assay (ELISA). Translocation of HMGB1 from the nucleus to the cytoplasm in macrophages was observed by Western blotting and activity of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-?B) in the nucleus was detected using ELISA. HMGB1 mRNA expression levels increased significantly in the cell culture supernatant and in cells after 24 h of stimulating RAW264.7 cells with LPS (500 ng/mL). However, HMGB1 mRNA expression levels in the P2 and P3 groups, which received 500 ng/mL LPS with 25 or 50 ?mol/mL propofol, respectively, were significantly lower than those in the group receiving LPS stimulation (P

  7. Alkaline Phosphatase Protects Lipopolysaccharide-Induced Early Pregnancy Defects in Mice

    Science.gov (United States)

    Lei, Wei; Ni, Hua; Herington, Jennifer; Reese, Jeff; Paria, Bibhash C.

    2015-01-01

    Excessive cytokine inflammatory response due to chronic or superphysiological level of microbial infection during pregnancy leads to pregnancy complications such as early pregnancy defects/loss and preterm birth. Bacterial toxin lipopolysaccharide (LPS), long recognized as a potent proinflammatory mediator, has been identified as a risk factor for pregnancy complications. Alkaline phosphatase (AP) isozymes have been shown to detoxify LPS by dephosphorylation. In this study, we examined the role of alkaline phosphatase (AP) in mitigating LPS-induced early pregnancy complications in mice. We found that 1) the uterus prior to implantation and implantation sites following embryo implantation produce LPS recognition and dephosphorylation molecules TLR4 and tissue non-specific AP (TNAP) isozyme, respectively; 2) uterine TNAP isozyme dephosphorylates LPS at its sites of production; 3) while LPS administration following embryo implantation elicits proinflammatory cytokine mRNA levels at the embryo implantation sites (EISs) and causes early pregnancy loss, dephosphorylated LPS neither triggers proinflammatory cytokine mRNA levels at the EISs nor induces pregnancy complications; 4) AP isozyme supplementation to accelerate LPS detoxification attenuates LPS-induced pregnancy complications following embryo implantation. These findings suggest that a LPS dephosphorylation strategy using AP isozyme may have a unique therapeutic potential to mitigate LPS- or Gram-negative bacteria-induced pregnancy complications in at-risk women. PMID:25910276

  8. Adrenomedullin gene expression and levels in the cardiovascular system after treatment with lipopolysaccharide.

    Science.gov (United States)

    Li, Yuk-Yin; Cheung, Bernard M Y; Wong, Louisa Y F; Hwang, Isabel S S; Kumana, Cyrus R; Tang, Fai

    2005-04-01

    To study the effect of septicaemia, the temporal changes in tissue adrenomedullin (AM) and preproAM mRNA levels were studied in the heart and blood vessels after lipopolysaccharide (LPS) injection. Radioimmunoassay and solution hybridization-RNase protection assays were used to follow the changes in AM and its mRNA levels respectively after intraperitoneal injection of 10 mg/kg LPS in rats. The preproAM mRNA levels increased at 1 h in the right atrium after LPS injection, while the AM contents decreased at 1 h in the left atrium. The preproAM mRNA levels increased at 3 and 6 h in the left ventricle, whereas it increased at 6 h in the right ventricles after LPS injection. There was an increase in preproAM mRNA levels at 1 and 3 h in the mesenteric artery, while AM levels were increased at 1, 3 and 6 h. However, there were no such changes in the thoracic aorta. There were also increases in tumour necrosis factor-alpha (TNF-alpha), interleukin (IL)-1beta and IL-6 in the heart, and in the mesenteric artery (TNF-alpha and IL-1beta) and in thoracic aorta (IL-1beta and IL-6). The present results suggest that the biosynthesis and secretion of AM may be increased in cardiovascular tissues of rats injected with LPS, and that AM may play multiple roles in inflammation. PMID:15752540

  9. Associations of Escherichia coli K-12 OmpF trimers with rough and smooth lipopolysaccharides

    International Nuclear Information System (INIS)

    The associations of both rough and smooth lipopolysaccharides (LPS) with the OmpF porin of Escherichia coli K-12 were examined in galE strains deleted for ompC. Transformation with pSS37 and growth with galactose conferred the ability to assemble a Shigella dysenteriae O antigen onto the core oligosaccharide of E. coli K-12 LPS. The association of LPS with OmpF trimers was assessed by staining, autoradiography of LPS specifically labeled with [1-14C]galactose, and Western immunoblotting with a monoclonal antibody specific for OmpF trimers. These techniques revealed that the migration distances and multiple banding patterns of OmpF porin trimers in sodium dodecyl sulfate-polyacrylamide gels were dictated by the chemotype of associated LPS. Expression of smooth LPS caused almost all of the trimeric OmpF to run in gels with a slower mobility than trimers from rough strains. The LPS associated with trimers from a smooth strain differed from the bulk-phase LPS by consisting almost exclusively of molecules with O antigen

  10. Recognition of bacterial lipopolysaccharide using bacteriophage-adhesin-coated long-period gratings.

    Science.gov (United States)

    Brzozowska, Ewa; ?mietana, Mateusz; Koba, Marcin; Górska, Sabina; Pawlik, Krzysztof; Gamian, Andrzej; Bock, Wojtek J

    2015-05-15

    In this paper we present a new type of highly sensitive label-free sensor based on long-period gratings (LPG) coated with T4 bacteriophage (phage) adhesin. The adhesin (gp37) binds Escherichia coli B (E. coli B) by recognizing its bacterial lipopolysaccharide (LPS). The LPG biofunctionalization methodology is based on coating LPG surface with nickel ions capable of gp37 histidine tag reversible binding. For the first time recombinant adhesive phage protein has been used as a receptor molecule in biosensing scheme. The specificity of LPS binding by adhesin has been tested with LPG-based device and confirmed using Western blot, Enzyme-Linked Immunosorbent Assay (ELISA) and BIACORE methods. The LPG-based sensor can measure bacterial contamination in real time and with a high accuracy. We show that T4 phage adhesin binds E. coli B LPS in its native or denatured form. The binding is highly specific and irreversible. The applied procedure allows for obtaining reusable biosensors. PMID:25067838

  11. Dried Ginger (Zingiber officinalis) Inhibits Inflammation in a Lipopolysaccharide-Induced Mouse Model

    Science.gov (United States)

    Choi, You Yeon; Kim, Mi Hye; Hong, Jongki; Kim, Sung-Hoon

    2013-01-01

    Objectives. Ginger rhizomes have a long history of human use, especially with regards to their anti-inflammatory properties. However, the mechanisms by which ginger acts on lipopolysaccharide-(LPS-)induced inflammation have not yet been identified. We investigated the anti-inflammatory effects of dried Zingiber officinalis (DZO) on LPS-induced hepatic injury. Methods. ICR mice were given a DZO water extract (100, 1000?mg/kg) orally for three consecutive days. On the third day, they were administered by LPS intraperitoneally. To investigate the anti-inflammatory effects of DZO, histological, cytokine expression, and protein factor analyses were performed. Results. Oral administration of DZO significantly reduced pathological changes in the liver and proinflammatory cytokines including interferon-(IFN-)? and interleukin-(IL-)6 in the serum. In addition, DZO inhibited LPS-induced NF-?B activation by preventing degradation of the I?B-?, as well as the phosphorylation of ERK1/2, SAPK/JNK, and p38 MAPKs. These were associated with a decrease in the expression of inducible nitric oxide synthase (iNOS) and cyclooxyenase-2 (COX-2). Conclusions. Our data provide evidence for the hepatoprotective mechanisms of DZO as an anti-inflammatory effect. Furthermore, use of DZO to treat could provide therapeutic benefits in clinical settings. PMID:23935687

  12. Intestinal alkaline phosphatase deficiency leads to lipopolysaccharide desensitization and faster weight gain.

    Science.gov (United States)

    Yang, Ye; Millán, José Luis; Mecsas, Joan; Guillemin, Karen

    2015-01-01

    Animals develop in the presence of complex microbial communities, and early host responses to these microbes can influence key aspects of development, such as maturation of the immune system, in ways that impact adult physiology. We previously showed that the zebrafish intestinal alkaline phosphatase (ALPI) gene alpi.1 was induced by Gram-negative bacterium-derived lipopolysaccharide (LPS), a process dependent on myeloid differentiation primary response gene 88 (MYD88), and functioned to detoxify LPS and prevent excessive host inflammatory responses to commensal microbiota in the newly colonized intestine. In the present study, we examined whether the regulation and function of ALPI were conserved in mammals. We found that among the mouse ALPI genes, Akp3 was specifically upregulated by the microbiota, but through a mechanism independent of LPS or MYD88. We showed that disruption of Akp3 did not significantly affect intestinal inflammatory responses to commensal microbiota or animal susceptibility to Yersinia pseudotuberculosis infection. However, we found that Akp3(-/-) mice acquired LPS tolerance during postweaning development, suggesting that Akp3 plays an important role in immune education. Finally, we demonstrated that inhibiting LPS sensing with a mutation in CD14 abrogated the accelerated weight gain in Akp3(-/-) mice receiving a high-fat diet, suggesting that the weight gain is caused by excessive LPS in Akp3(-/-) mice. PMID:25348635

  13. Binding interactions of bacterial lipopolysaccharide and the cationic amphiphilic peptides polymyxin B and WLBU2.

    Science.gov (United States)

    Ryder, Matthew P; Wu, Xiangming; McKelvey, Greg R; McGuire, Joseph; Schilke, Karl F

    2014-08-01

    Passage of blood through a sorbent device for removal of bacteria and endotoxin by specific binding with immobilized, membrane-active, bactericidal peptides holds promise for treating severe blood infections. Peptide insertion in the target membrane and rapid/strong binding is desirable, while membrane disruption and release of degradation products to the circulating blood is not. Here we describe interactions between bacterial endotoxin (lipopolysaccharide, LPS) and the membrane-active, bactericidal peptides WLBU2 and polymyxin B (PmB). Analysis of the interfacial behavior of mixtures of LPS and peptide using air-water interfacial tensiometry and optical waveguide lightmode spectroscopy strongly suggests insertion of intact LPS vesicles by the peptide WLBU2 without vesicle destabilization. In contrast, dynamic light scattering (DLS) studies show that LPS vesicles appear to undergo peptide-induced destabilization in the presence of PmB. Circular dichroism spectra further confirm that WLBU2, which shows disordered structure in aqueous solution and substantially helical structure in membrane-mimetic environments, is stably located within the LPS membrane in peptide-vesicle mixtures. We therefore expect that presentation of WLBU2 at an interface, if tethered in a fashion which preserves its mobility and solvent accessibility, will enable the capture of bacteria and endotoxin without promoting reintroduction of endotoxin to the circulating blood, thus minimizing adverse clinical outcomes. On the other hand, our results suggest no such favorable outcome of LPS interactions with polymyxin B. PMID:24905681

  14. Astragalus mongholicus polysaccharide inhibits lipopolysaccharide-induced production of TNF-? and interleukin-8

    Science.gov (United States)

    Yuan, Yuan; Sun, Mei; Li, Ke-Shen

    2009-01-01

    AIM: To explore the effect of Astragalus mongholicus polysaccharide (APS) on gene expression and mitogen-activated protein kinase (MAPK) transcriptional activity in intestinal epithelial cells (IEC). METHODS: IEC were divided into control group, lipopolysaccharide (LPS) group, LPS+ 50 ?g/mL APS group, LPS+ 100 ?g/mL APS group, LPS+ 200 ?g/mL APS group, and LPS+ 500 ?g/mL APS group. Levels of mRNAs in LPS-induced inflammatory factors, tumor necrosis factor (TNF)-? and interleukin (IL)-8, were measured by reverse transcription-polymerase chain reaction. MAPK protein level was measured by Western blotting. RESULTS: The levels of TNF-? and IL-8 mRNAs were significantly higher in IEC with LPS-induced damage than in control cells. APS significantly abrogated the LPS-induced expression of the TNF-? and IL-8 genes. APS did not block the activation of extracellular signal-regulated kinase or c Jun amino-terminal kinase, but inhibited the activation of p38, suggesting that APS inhibits LPS-induced production of TNF-? and IL-8 mRNAs, possibly by suppressing the p38 signaling pathway. CONCLUSION: APS-modulated bacterial product-mediated p38 signaling represents an attractive strategy for prevention and treatment of intestinal inflammation. PMID:19653348

  15. Priming, induction and modulation of plant defence responses by bacterial lipopolysaccharides

    DEFF Research Database (Denmark)

    Newman, Mari-Anne; Dow, J. Maxwell

    2007-01-01

    Bacterial lipopolysaccharides (LPSs) have multiple roles in plant-microbe interactions. LPS contributes to the low permeability of the outer membrane, which acts as a barrier to protect bacteria from plant-derived antimicrobial substances. Conversely, perception of LPS by plant cells can lead to the triggering of defence responses or to the priming of the plant to respond more rapidly and/or to a greater degree to subsequent pathogen challenge. LPS from symbiotic bacteria can have quite different effects on plants to those of pathogens. Some details are emerging of the structures within LPS that are responsible for induction of these different plant responses. The lipid A moiety is not solely responsible for all of the effects of LPS in plants; core oligosaccharide and O-antigen components can elicit specific responses. Here, we review the effects of LPS in induction of defence-related responses in plants, the structures within LPS responsible for eliciting these effects and discuss the possible nature of the(as yet unidentified) LPS receptors in plants.

  16. Defects in rhizobial cyclic glucan and lipopolysaccharide synthesis alter legume gene expression during nodule development

    DEFF Research Database (Denmark)

    D'Antuono, Alejandra L; Ott, Thomas

    2008-01-01

    cDNA array technology was used to compare transcriptome profiles of Lotus japonicus roots inoculated with a Mesorhizobium loti wild-type and two mutant strains affected in cyclic beta(1-2) glucan synthesis (cgs) and in lipopolysaccharide synthesis (lpsbeta2). Expression of genes associated with the development of a fully functional nodule was significantly affected in plants inoculated with the cgs mutant. Array results also revealed that induction of marker genes for nodule development was delayed when plants were inoculated with the lpsbeta2 mutant. Quantitative real-time reverse-transcriptase polymerase chain reaction was used to quantify gene expression of a subset of genes involved in plant defense response, redox metabolism, or genes that encode for nodulins. The majority of the genes analyzed in this study were more highly expressed in roots inoculated with the wild type compared with those inoculated with the cgs mutant strain. Some of the genes exhibited a transient increase in transcript levels during intermediate steps of normal nodule development while others displayed induced expression during the final steps of nodule development. Ineffective nodules induced by the glucan mutant showed higher expression of phenylalanine ammonia lyase than wild-type nodules. Differences in expression pattern of genes involved in early recognition and signaling were observed in plants inoculated with the M. loti mutant strain affected in the synthesis of cyclic glucan. Udgivelsesdato: 2008-Jan

  17. The effects of propolis on cytokine production in lipopolysaccharide-stimulated macrophages

    Directory of Open Access Journals (Sweden)

    Hatice Özbilge

    2011-12-01

    Full Text Available Objectives: Propolis, a bee-product, has attracted researchers’ interest in recent years because of several biological and pharmacological properties. Lipopolysaccharide (LPS is a component of the outer membrane of Gram-negative bacteria and has an important role in the pathogenesis of septic shock and several inflammatory diseases by causing excessive release of inflammatory cytokines. The aim of this study was to investigate the effects of ethanol extract of propolis collected in Kayseri and its surroundings on production of pro-inflammatory cytokines in LPS-stimulated macrophages.Materials and methods: In vitro, U937 human macrophage cells were grown in RPMI-1640 medium supplemented with fetal bovine serum (10% and penicillin-streptomycin (2% and divided into: control, LPS treated, and propolis+LPS treated cell groups. After incubation in an atmosphere of 5% CO2 and at 37°C of cells, interleukin (IL-1?, IL-6 and tumor necrosis factor (TNF-? levels were measured in cell-free supernatants by ELISA.Results: IL-1?, IL-6 and TNF-? levels increased in LPS treated cell group according to control, statistically significant. Each cytokine levels significantly decreased in LPS and propolis treated cell group according to only LPS treated cell group (p<0.05.Conclusion: Propolis is a natural product to be examined for usage when needed the suppression of pro-inflammatory cytokines. J Clin Exp Invest 2011; 2 (4: 366-370

  18. Lipopolysaccharide of Marinobacter litoralis inhibits swarming motility and biofilm formation in Pseudomonas aeruginosa PA01.

    Science.gov (United States)

    Sardar, Raj Kumar; Kavita, Kumari; Jha, Bhavanath

    2015-06-01

    The lipopolysaccharide (LPS) was isolated from a marine bacterium identified as Marinobacter litoralis BK09 using 16S rRNA gene sequence similarity analysis. The GCMS analysis showed that the LPS contained 3-hydroxy-dodecanoic acid (C12:0 3OH) (49%), dodecanoic acid (C12:0) (24%) and decanoic acid (C10:0) (19%) as major fatty acids, and the polysaccharide constituents were fucose (53.79%), xylose (28.04%) and mannose (18.15%). The LPS almost completely inhibited swarming motility in Pseudomonas aeruginosa PA01. It also reduced biofilm formation by 50% with no adverse effect on cell growth. The production of virulence factor such as pyocyanin pigment was reduced (?40%) by the LPS. The LPS did not show any limulus amoebocyte lysate (LAL) gelation activity. The repression of swarming motility, pyocyanin production and biofilm formation by the LPS suggests its potential application against P. aeruginosa infection. This is the first report on characterization and application of LPS from M. litoralis. PMID:25843881

  19. Characterization of monoclonal antibodies to terminal and internal O-antigen epitopes of Francisella tularensis lipopolysaccharide.

    Science.gov (United States)

    Roche, Marly I; Lu, Zhaohua; Hui, Julia H; Sharon, Jacqueline

    2011-02-01

    The lipopolysaccharide (LPS) of Francisella tularensis (Ft), the Gram negative bacterium that causes tularemia, has been shown to be a main protective antigen in mice and humans; we have previously demonstrated that murine anti-Ft LPS IgG2a monoclonal antibodies (MAbs) can protect mice against otherwise lethal intranasal infection with the Ft live vaccine strain (LVS). Here we show that four IgG2a anti-LPS MAbs are specific for the O-polysaccharide (O-antigen [OAg]) of Ft LPS. But whereas three of the MAbs bind to immunodominant repeating internal epitopes, one binds to a unique terminal epitope of Ft OAg. This was deduced from its even binding to both long and short chains of the LPS ladder in Western blots, its rapid decrease in ELISA binding to decreasing solid-phase LPS concentrations, its inability to compete for LPS binding with a representative of the other three MAbs, and its inability to immunoprecipitate OAg despite its superior agglutination titer. Biacore analysis showed the end-binding MAb to have higher bivalent avidity for Ft OAg than the internal-binding MAbs and provided an immunogenicity explanation for the predominance of internal-binding anti-Ft OAg MAbs. These findings demonstrate that non-overlapping epitopes can be targeted by antibodies to Ft OAg, which may inform the design of vaccines and immunotherapies against tularemia. PMID:21466282

  20. Molecular and chemical analysis of the lipopolysaccharide from aeromonas hydrophila strain AH-1 (Serotype O11).

    Science.gov (United States)

    Merino, Susana; Canals, Rocío; Knirel, Yuriy A; Tomás, Juan M

    2015-04-01

    A group of virulent Aeromonas hydrophila, A. sobria, and A. veronii biovar sobria strains isolated from humans and fish have been described; these strains classified to serotype O11 are serologically related by their lipopolysaccharide (LPS) O-antigen (O-polysaccharide), and the presence of an S-layer consisting of multiple copies of a crystalline surface array protein with a molecular weight of 52 kDa in the form of a crystalline surface array which lies peripheral to the cell wall. A. hydrophila strain AH-1 is one of them. We isolated the LPS from this strain and determined the structure of the O-polysaccharide, which was similar to that previously described for another strain of serotype O11. The genetics of the O11-antigen showed the genes (wbO11 cluster) in two sections separated by genes involved in biosynthesis and assembly of the S-layer. The O11-antigen LPS is an example of an ABC-2-transporter-dependent pathway for O-antigen heteropolysaccharide (disaccharide) assembly. The genes involved in the biosynthesis of the LPS core (waaO11 cluster) were also identified in three different chromosome regions being nearly identical to the ones described for A. hydrophila AH-3 (serotype O34). The genetic data and preliminary chemical analysis indicated that the LPS core for strain AH-1 is identical to the one for strain AH-3. PMID:25874921

  1. Effects of propofol on damage of rat intestinal epithelial cells induced by heat stress and lipopolysaccharides

    Scientific Electronic Library Online (English)

    J., Tang; Y., Jiang; Y., Tang; B., Chen; X., Sun; L., Su; Z., Liu.

    2013-06-25

    Full Text Available Gut-derived endotoxin and pathogenic bacteria have been proposed as important causative factors of morbidity and death during heat stroke. However, it is still unclear what kind of damage is induced by heat stress. In this study, the rat intestinal epithelial cell line (IEC-6) was tre [...] ated with heat stress or a combination of heat stress and lipopolysaccharide (LPS). In addition, propofol, which plays an important role in anti-inflammation and organ protection, was applied to study its effects on cellular viability and apoptosis. Heat stress, LPS, or heat stress combined with LPS stimulation can all cause intestinal epithelial cell damage, including early apoptosis and subsequent necrosis. However, propofol can alleviate injuries caused by heat stress, LPS, or the combination of heat stress and LPS. Interestingly, propofol can only mitigate LPS-induced intestinal epithelial cell apoptosis, and has no protective role in heat-stress-induced apoptosis. This study developed a model that can mimic the intestinal heat stress environment. It demonstrates the effects on intestinal epithelial cell damage, and indicated that propofol could be used as a therapeutic drug for the treatment of heat-stress-induced intestinal injuries.

  2. Activation of PPAR? by Wy-14643 ameliorates systemic lipopolysaccharide-induced acute lung injury

    Energy Technology Data Exchange (ETDEWEB)

    Yoo, Seong Ho, E-mail: yoosh@snu.ac.kr [Seoul National University Hospital, Biomedical Research Institute and Institute of Forensic Medicine, Seoul National University College of Medicine, Seoul (Korea, Republic of); Abdelmegeed, Mohamed A. [Laboratory of Membrane Biochemistry and Biophysics, National Institute on Alcohol Abuse and Alcoholism, Bethesda, MD (United States); Song, Byoung-Joon, E-mail: bj.song@nih.gov [Laboratory of Membrane Biochemistry and Biophysics, National Institute on Alcohol Abuse and Alcoholism, Bethesda, MD (United States)

    2013-07-05

    Highlights: •Activation of PPAR? attenuated LPS-mediated acute lung injury. •Pretreatment with Wy-14643 decreased the levels of IFN-? and IL-6 in ALI. •Nitrosative stress and lipid peroxidation were downregulated by PPAR? activation. •PPAR? agonists may be potential therapeutic targets for acute lung injury. -- Abstract: Acute lung injury (ALI) is a major cause of mortality and morbidity worldwide. The activation of peroxisome proliferator-activated receptor-? (PPAR?) by its ligands, which include Wy-14643, has been implicated as a potential anti-inflammatory therapy. To address the beneficial efficacy of Wy-14643 for ALI along with systemic inflammation, the in vivo role of PPAR? activation was investigated in a mouse model of lipopolysaccharide (LPS)-induced ALI. Using age-matched Ppara-null and wild-type mice, we demonstrate that the activation of PPAR? by Wy-14643 attenuated LPS-mediated ALI. This was evidenced histologically by the significant alleviation of inflammatory manifestations and apoptosis observed in the lung tissues of wild-type mice, but not in the corresponding Ppara-null mice. This protective effect probably resulted from the inhibition of LPS-induced increases in pro-inflammatory cytokines and nitroxidative stress levels. These results suggest that the pharmacological activation of PPAR? might have a therapeutic effect on LPS-induced ALI.

  3. The structures of lipopolysaccharides from plant-associated gram-negative bacteria

    DEFF Research Database (Denmark)

    Molinaro, Antonio; Newman, Mari-Anne

    2009-01-01

    Gram-negative bacterial lipopolysaccharides (LPSs) have multiple roles in plant-microbe interactions. LPSs contribute to the low permeabilities of bacterial outer membranes, which act as barriers to protect bacteria from plant-derived antimicrobial substances. Conversely, perception of LPSs by plant cells can lead to the triggering of defence responses or to the priming of the plant to respond more rapidly and/or to a greater degree to subsequent pathogen challenge. LPSs are thus key molecules in the interactions between bacteria and plants, either in symbiosis or pathogenesis. Since LPSs are glycoconjugates genetically and chemically consisting of three different molecular regions, their detailed structure elucidation is a very topical and major scientific task for chemists, and is achieved by a combination of state-of-art chemical and spectroscopic techniques. Knowledge of LPSs' chemical structures is an important prerequisite for any further understanding of the biological processes in plant-microbe interactions. Moreover, the LPSs from Gram-negative bacteria - especially those originating from plant-associated bacteria - are a great source of novel monosaccharides with unusual and occasionally astounding chemical structures, never found in the eukaryotic world. This review presents the structures of LPSs from plant-associated bacteria isolated and identified from 2001 onwards. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2009)

  4. The rapid lipopolysaccharide-induced release of matrix metalloproteinases 9 is suppressed by simvastatin.

    Science.gov (United States)

    Li, Dan-Dong; Pang, Hong-Gang; Song, Jin-Ning; Huang, Huan; Zhang, Ming; Zhao, Yong-Lin; Sun, Peng; Zhang, Bin-Fei; Ma, Xu-Dong

    2015-07-01

    A rapid increase in matrix metalloproteinase-9 (MMP-9) expression by stimulated leukocytes is common in many diseases. Recent evidence suggests that the beneficial effects of statins are mediated in part by the suppression of MMP-9 release. In this study, we investigated the effect of statin on MMP-9 expression and its antagonist, tissue inhibitor of metalloproteinase-1 (TIMP-1) in LPS-stimulated leukocytes. Rat neutrophils and monocytes were stimulated with lipopolysaccharide (LPS) in the presence of simvastatin. MMP-9 secretion and mRNA expression were analyzed using ELISA and RT-PCR, respectively. Total MMP-9 protein production was measured by Western blot analysis. Potential signal transduction pathways responsible for MMP-9 production were investigated using luciferase reporter assays (NF-?B), pull-down assays (RhoA), and pharmacological inhibition. Our data show that MMP-9 and TIMP-1 expression are differentially induced by LPS in neutrophils and monocytes. We showed that rapid MMP-9 release occurred mainly via secretion from intracellular stores. Moreover, we showed that statin significantly suppressed LPS-induced MMP-9 release and mRNA expression in a time- and concentration-dependent manner. We also evaluated that simvastain postponed the rapid LPS-induced MMP-9 release for about 20?min. In conclusion, we demonstrated that the suppressive effect of simvastatin on LPS-stimulated MMP-9 release does not occur via the NF-?B pathway and the MAPKs pathway, but via the RhoA/ROCK pathway. PMID:25612169

  5. Astragaloside IV prevents lipopolysaccharide-induced injury in H9C2 cardiomyocytes.

    Science.gov (United States)

    Wang, Shi-Guang; Xu, Yan; Xie, Hao; Wang, Wei; Chen, Xiao-Hu

    2015-02-01

    This study aimed to investigate the protective effects of astragaloside IV (AS IV) on lipopolysaccharide (LPS)-induced injury in H9C2 cardiomyocytes. H9C2 Cardiomyocytes were cultured with LPS (10 ?g·mL(-1)) for 4 h and treated with AS IV at 50, 100, and 150 ?mol·L(-1) for various durations. Cell viability was determined by MTT. The content of released TNF-? and IL-6 from cardiomyocytes were evaluated by enzyme-linked immunosorbent assay (ELISA). The levels of superoxidase dismutase (SOD), malondialdehyde (MDA), lactate dehydrogenase (LDH), and creatine phosphate kinase (CK) were measured by using commercial available kits. The mRNA and protein expression levels of NF-?B p65 were measured by RT-PCR and Western blotting, respectively. And the NF-?B p65 activity was measured by ELISA. Our results demonstrated that AS IV at 50, 100, and 150 ?mol·L(-1) markedly inhibited the release of TNF-? and IL-6 and decreased NF-?B expression, compared with the model group. Moreover, the improved SOD activity and decreased MDA, LDH and CK levels were detected after AS IV treatment. In summary, AS IV could increase the activities of antioxidant enzymes, inhibite lipid peroxidation, and down-regulate the inflammatory mediators involved in the inflammatory responses. These results demonstrated that AS IV could prevent LPS-induced injury in cardiomyocytes. PMID:25769895

  6. Epitope recognition of antibodies against a Yersinia pestis lipopolysaccharide trisaccharide component.

    Science.gov (United States)

    Broecker, Felix; Aretz, Jonas; Yang, You; Hanske, Jonas; Guo, Xiaoqiang; Reinhardt, Anika; Wahlbrink, Annette; Rademacher, Christoph; Anish, Chakkumkal; Seeberger, Peter H

    2014-04-18

    Today, the process of selecting carbohydrate antigens as a basis for active vaccination and the generation of antibodies for therapeutic and diagnostic purposes is based on intuition combined with trial and error experiments. In efforts to establish a rational process for glycan epitope selection, we employed glycan array screening, surface plasmon resonance, and saturation transfer difference (STD)-NMR to elucidate the interactions between antibodies and glycans representing the Yersinia pestis lipopolysaccharide (LPS). A trisaccharide epitope of the LPS inner core glycan and different LPS-derived oligosaccharides from various Gram-negative bacteria were analyzed using this combination of techniques. The antibody-glycan interaction with a heptose substructure was determined at atomic-level detail. Antibodies specifically recognize the Y. pestis trisaccharide and some substructures with high affinity and specificity. No significant binding to LPS glycans from other bacteria was observed, which suggests that the epitopes for just one particular bacterial species can be identified. On the basis of these results we are beginning to understand the rules for structure-based design and selection of carbohydrate antigens. PMID:24479563

  7. Ginsenoside Rg3 attenuates microglia activation following systemic lipopolysaccharide treatment in mice.

    Science.gov (United States)

    Park, Sun-Min; Choi, Moon-Suk; Sohn, Nak-Won; Shin, Jung-Won

    2012-01-01

    Neuroinflammation, characterized by activation of microglia and expression of major inflammatory mediators, contributes to neuronal damage in addition to acute and chronic central nervous system (CNS) disease progression. The present study investigated the immune modulatory effects of ginsenoside Rg3, a principle active ingredient in Panax ginseng, on pro-inflammatory cytokines and microglia activation in brain tissue induced by systemic lipopolysaccharide (LPS) treatment in C57BL/6 mice. Systemic LPS treatment induces immediate microglia activation in the brain. Based on this information, ginsenoside Rg3 was treated orally with 10, 20, and 30?mg/kg 1?h prior to the LPS (3?mg/kg, intraperitoneally (i.p.)) injection. Ginsenoside Rg3 at 20 and 30?mg/kg oral doses significantly attenuated up-regulation of tumor necrosis factor-alpha (TNF-?), interleukin-1 beta (IL-1?) and IL-6 mRNA in brain tissue at 4?h after LPS injection. Morphological activation of microglia and Iba1 protein expression by systemic LPS injection were reduced with ginsenoside Rg3 (30?mg/kg) treatment. In addition, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression in brain tissue were also attenuated with oral treatment of ginsenoside Rg3 at 30?mg/kg. These results indicate that ginsenoside Rg3 plays a modulatory role in neuroinflammation. This study shows that ginsenoside Rg3 attenuates microglia activation using an in vivo animal model. PMID:22975507

  8. Lipopolysaccharide induces multinuclear cell from RAW264.7 line with increased phagocytosis activity

    International Nuclear Information System (INIS)

    Highlights: ? LPS induces multinuclear cells from murine macrophage-derived RAW264.7 cells. ? The multinuclear cells are formed through cell-cell fusion in the presence of Ca2+. ? The multinuclear cells do not express osteoclast-specific enzymes. ? They internalized more and larger beads than mononuclear cells and osteoclasts. -- Abstract: Lipopolysaccharide (LPS), an outer membrane component of Gram-negative bacteria, induces strong proinflammatory responses, including the release of cytokines and nitric oxide from macrophage. In this study, we found that a murine macrophage-derived line, RAW264.7, became multinuclear through cell-cell fusion after incubation with highly purified LPS or synthetic lipid A in the presence of Ca2+. The same cell line is known to differentiate into multinuclear osteoclast, which expresses a specific proton pumping ATPase together with osteoclast markers on stimulation by the extracellular domain of receptor activator of nuclear factor ?B ligand (Toyomura, T., Murata, Y., Yamamoto, A., Oka, T., Sun-Wada, G.-H., Wada, Y. and Futai, M., 2003). The LPS-induced multinuclear cells did not express osteoclast-specific enzymes including tartrate-resistant acid phosphatase and cathepsin K. During multinuclear cell formation, the cells internalized more and larger polystyrene beads (diameter 6–15 ?m) than mononuclear cells and osteoclasts. The internalized beads were located in lysosome-marker positive organelles, which were probably phagolysosomes. The LPS-induced multinuclear cell could be a good model system to study phagocytosis of large foreign bodies.

  9. Activation of PPAR? by Wy-14643 ameliorates systemic lipopolysaccharide-induced acute lung injury

    International Nuclear Information System (INIS)

    Highlights: •Activation of PPAR? attenuated LPS-mediated acute lung injury. •Pretreatment with Wy-14643 decreased the levels of IFN-? and IL-6 in ALI. •Nitrosative stress and lipid peroxidation were downregulated by PPAR? activation. •PPAR? agonists may be potential therapeutic targets for acute lung injury. -- Abstract: Acute lung injury (ALI) is a major cause of mortality and morbidity worldwide. The activation of peroxisome proliferator-activated receptor-? (PPAR?) by its ligands, which include Wy-14643, has been implicated as a potential anti-inflammatory therapy. To address the beneficial efficacy of Wy-14643 for ALI along with systemic inflammation, the in vivo role of PPAR? activation was investigated in a mouse model of lipopolysaccharide (LPS)-induced ALI. Using age-matched Ppara-null and wild-type mice, we demonstrate that the activation of PPAR? by Wy-14643 attenuated LPS-mediated ALI. This was evidenced histologically by the significant alleviation of inflammatory manifestations and apoptosis observed in the lung tissues of wild-type mice, but not in the corresponding Ppara-null mice. This protective effect probably resulted from the inhibition of LPS-induced increases in pro-inflammatory cytokines and nitroxidative stress levels. These results suggest that the pharmacological activation of PPAR? might have a therapeutic effect on LPS-induced ALI

  10. Intranasal curcumin ameliorates lipopolysaccharide-induced acute lung injury in mice.

    Science.gov (United States)

    Kumari, Asha; Tyagi, Namitosh; Dash, D; Singh, Rashmi

    2015-06-01

    Lipopolysaccharide (LPS) is one of the most powerful proinflammatory factor and can induce acute pulmonary inflammation even lung injury after inhalation or systemic administration. LPS induces sepsis and multiple organ damage. Curcumin (diferuloylmethane), a major component of turmeric, exhibits protection against LPS-induced acute lung injury (ALI). We aimed to investigate effects of intranasal curcumin on LPS-induced ALI in mice where curcumin (10 mg/kg, intranasal (i.n.) was given an hour before LPS exposure. After 24 h of intranasal LPS instillation, a marked increase in neutrophil recruitment and myeloperoxidase (MPO) activity was noted which were significantly ameliorated in curcumin treatment group. Oxidative stress markers like nitric oxide (NO), malondialdehyde (MDA) level and evans blue capillary leakage assay also revealed suppression after curcumin treatment; interestingly, levels of anti-oxidative enzymes such as superoxide dismutase (SOD) and catalase were upregulated. Inflammatory cytokine, tumour necrosis factor alpha (TNF-?) level was significantly attenuated by curcumin. Hence, intranasal curcumin could be a novel therapeutic strategy for LPS-induced ALI by directly targeting the lungs and enhancing anti-oxidant levels. PMID:25526714

  11. Protection against Lipopolysaccharide-induced Death by An Anti-Interleukin-6 Monoclonal Antibody

    Directory of Open Access Journals (Sweden)

    Bailin Liang

    2007-01-01

    Full Text Available Septicemia is frequently associated with serious complications and high mortality despite recent advances in treatment with antibacterial agents. Infectious organisms or their soluble products initiate septic shock through a complex cytokine cascade. Lipopolysaccharide (LPS induced excessive production of inflammatory cytokines is regarded as a model of septic shock. Experiments of LPS-induced excessive cytokine production were conducted in Balb/c mice for the investigation of the therapeutic potential of anti-TNF-? and anti-IL-6 monoclonal antibody treatments. LPS injection resulted in mortality with significant serum levels of TNF-?, IL-6, and IL-1?. Anti-IL-6 treatment significantly reduced animal mortality and inhibited serum levels of IL-6. Anti-TNF-? treatment did not affect serum IL-6 levels even though it significantly enhanced animal survival and inhibited TNF-? production. Moreover, anti-IL-6 treatment modestly reduced serum TNF-α level in comparison with control antibody treated group. These data suggest that TNF-? and IL-6 may play distinct protective roles in septic shock.

  12. Anti-inflammatory effects of a methanol extract from Pulsatilla koreana in lipopolysaccharide-exposed rats

    Directory of Open Access Journals (Sweden)

    Sang Hyun Lee1, Eun Lee1,* & Young Tag Ko2,*

    2012-06-01

    Full Text Available To investigate the therapeutic effect of a Korean herbal medicinePulsatilla koreana as an anti-septic agent, anti-inflammatoryeffects of the herbal medicine were determined in lipopolysaccharide(LPS-exposed rats. Treatment with a methanol extractfrom Pulsatilla koreana significantly inhibited LPS-inducedinflammatory responses. Results from ELISA analysis showed thatPulsatilla koreana decreased the plasma and hepatic levels ofpro-inflammatory cytokines such as IL-1?, IL-6, TNF-? whileincreased the level of anti-inflammatory cytokine IL-10 inLPS-exposed rats. Pulsatilla koreana also decreased the plasmalevels of other inflammatory mediators such as NO3-/NO2-,ICAM-1, PGE2, and CINC-1 in LPS-exposed rats. Although nosignificant effects were observed in the phagocytic activities, thedistribution of lymphocyte population was significantly shiftedby the treatment with Pulsatilla koreana. All together, Pulsatillakoreana exerts anti-inflammatory activities in the immunechallengedanimals implicating that this Korean herbal medicineis therapeutically useful for the treatment of inflammatorydiseases like sepsis.

  13. Alcohol metabolites and lipopolysaccharide: Roles in the development and/or progression of alcoholic liver disease

    Directory of Open Access Journals (Sweden)

    Courtney S Schaffert, Michael J Duryee, Carlos D Hunter, Bartlett C Hamilton 3rd, Amy L DeVeney, Mary M Huerter, Lynell W Klassen, Geoffrey M Thiele

    2009-03-01

    Full Text Available The onset of alcoholic liver disease (ALD is initiated by different cell types in the liver and a number of different factors including: products derived from ethanol-induced inflammation, ethanol metabolites, and the indirect reactions from those metabolites. Ethanol oxidation results in the production of metabolites that have been shown to bind and form protein adducts, and to increase inflammatory, fibrotic and cirrhotic responses. Lipopolysaccharide (LPS has many deleterious effects and plays a significant role in a number of disease processes by increasing inflammatory cytokine release. In ALD, LPS is thought to be derived from a breakdown in the intestinal wall enabling LPS from resident gut bacterial cell walls to leak into the blood stream. The ability of adducts and LPS to independently stimulate the various cells of the liver provides for a two-hit mechanism by which various biological responses are induced and result in liver injury. Therefore, the purpose of this article is to evaluate the effects of a two-hit combination of ethanol metabolites and LPS on the cells of the liver to increase inflammation and fibrosis, and play a role in the development and/or progression of ALD.

  14. Chemical characterization and biological properties of lipopolysaccharides isolated from smooth and rough strains of Brucella abortus.

    Science.gov (United States)

    Kreutzer, D L; Buller, C S; Robertson, D C

    1979-01-01

    The chemical composition and some of the biological activities of lipopolysaccharides (LPS) extracted from a smooth-intermediate strain (45/0) and a rough strain (45/20) of Brucella abortus have been examined. LPS were found in both the phenolic and aqueous extraction phases of strain 45/0, but only in the aqueous phase of 45/20. The phenolic LPS contained 9- to 16-fold-lower levels of heptose and reduced amounts of dideoxyaldoses compared with aqueous fractions. The major neutral sugars were glucose, galactose, and mannose. beta-Hydroxymyristic-acid, a common marker of enteric LPS, was not detected. Fatty acids present in highest amounts were hydroxylated and nonhydroxylated species with chain lengths of 16, 18, and 20 carbons. Only the phenolic LPS of strain 45/0 exhibited mouse lethality and a curable wasting disease; however, both phenolic and aqueous fractions caused carbohydrate depletion in mice. The toxicity of aqueous LPS could not be potentiated with Pb(OAc)4. These data, coupled with the lack of mitogenic activity for B-lymphocytes, are indicative of the unique structure-function relationships of Brucella LPS. Images PMID:110682

  15. Heat shock inhibits lipopolysaccharide-induced tissue factor activity in human whole blood

    Directory of Open Access Journals (Sweden)

    Thielmann Matthias

    2007-09-01

    Full Text Available Abstract Background During gram-negative sepsis, lipopolysaccharide (LPS induces tissue factor expression on monocytes. The resulting disseminated intravascular coagulation leads to tissue ischemia and worsens the prognosis of septic patients. There are indications, that fever reduces the mortality of sepsis, the effect on tissue factor activity on monocytes is unknown. Therefore, we investigated whether heat shock modulates LPS-induced tissue factor activity in human blood. Methods Whole blood samples and leukocyte suspensions, respectively, from healthy probands (n = 12 were incubated with LPS for 2 hours under heat shock conditions (43°C or control conditions (37°C, respectively. Subsequent to further 3 hours of incubation at 37°C the clotting time, a measure of tissue factor expression, was determined. Cell integrity was verified by trypan blue exclusion test and FACS analysis. Results Incubation of whole blood samples with LPS for 5 hours at normothermia resulted in a significant shortening of clotting time from 357 ± 108 sec to 82 ± 8 sec compared to samples incubated without LPS (n = 12; p 0.05. Similarly, heat shock treatment of leukocyte suspensions abolished the LPS-induced tissue factor activity. Clotting time was 73 ± 31 s, when cells were treated with LPS (100 ng/mL under normothermic conditions, and 301 ± 118 s, when treated with LPS (100 ng/mL and heat shock (n = 8, p Conclusion Heat shock treatment inhibits LPS-induced tissue factor activity in human whole blood samples and isolated leukocytes.

  16. Interactions of a designed peptide with lipopolysaccharide: Bound conformation and anti-endotoxic activity

    International Nuclear Information System (INIS)

    Designed peptides that would selectively interact with lipopolysaccharide (LPS) or endotoxin and fold into specific conformations could serve as important scaffolds toward the development of antisepsis compounds. Here, we describe solution structure of a designed amphipathic peptide, H2N-YVKLWRMIKFIR-CONH2 (YW12D) in complex with endotoxin as determined by transferred nuclear Overhauser effect spectroscopy. The conformation of the isolated peptide is highly flexible, but undergoes a dramatic structural stabilization in the presence of LPS. Structure calculations reveal that the peptide presents two amphipathic surfaces in its bound state to LPS whereby each surface is characterized by two positive charges and a number of aromatic and/or aliphatic residues. ITC data suggests that peptide interacts with two molecules of lipid A. In activity assays, YW12D exhibits neutralization of LPS toxicity with very little hemolysis of red blood cells. Structural and functional properties of YW12D would be applicable in designing low molecular weight non-toxic antisepsis molecules

  17. Early effects of lipopolysaccharide-induced inflammation on foetal brain development in rat

    Directory of Open Access Journals (Sweden)

    Cristina A Ghiani

    2011-11-01

    Full Text Available Studies in humans and animal models link maternal infection and imbalanced levels of inflammatory mediators in the foetal brain to the aetiology of neuropsychiatric disorders. In a number of animal models, it was shown that exposure to viral or bacterial agents during a period that corresponds to the second trimester in human gestation triggers brain and behavioural abnormalities in the offspring. However, little is known about the early cellular and molecular events elicited by inflammation in the foetal brain shortly after maternal infection has occurred. In this study, maternal infection was mimicked by two consecutive intraperitoneal injections of 200 ?g of LPS (lipopolysaccharide/kg to timed-pregnant rats at GD15 (gestational day 15 and GD16. Increased thickness of the CP (cortical plate and hippocampus together with abnormal distribution of immature neuronal markers and decreased expression of markers for neural progenitors were observed in the LPS-exposed foetal forebrains at GD18. Such effects were accompanied by decreased levels of reelin and the radial glial marker GLAST (glial glutamate transporter, and elevated levels of pro-inflammatory cytokines in maternal serum and foetal forebrains. Foetal inflammation elicited by maternal injections of LPS has discrete detrimental effects on brain development. The early biochemical and morphological changes described in this work begin to explain the sequelae of early events that underlie the neurobehavioural deficits reported in humans and animals exposed to prenatal insults.

  18. Tocotrienols inhibit lipopolysaccharide-induced pro-inflammatory cytokines in macrophages of female mice

    Directory of Open Access Journals (Sweden)

    Morrison David C

    2010-12-01

    Full Text Available Abstract Background Inflammation has been implicated in cardiovascular disease, and the important role of proteasomes in the development of inflammation and other macrophage functions has been demonstrated. Tocotrienols are potent hypocholesterolemic agents that inhibit ?-hydroxy-?-methylglutaryl coenzyme A reductase activity, which is degraded via the ubiquitin-proteasome pathway. Our objective was to evaluate the effect of tocotrienols in reducing inflammation. Lipopolysaccharide (LPS was used as a prototype for inflammation in murine RAW 264.7 cells and BALB/c female mice. Results The present results clearly demonstrate that ?-, ?-, or ?-tocotrienol treatments inhibit the chymotrypsin-like activity of 20 S rabbit muscle proteasomes (> 50%; P 40% (P P P P P 40%, while higher concentrations (40 ?M increased gene expression of the latter in peritoneal macrophages (prepared from BALB/c mice as compared to control group. Conclusions These results represent a novel approach by using natural products, such as tocotrienols as proteasome modulators, which may lead to the development of new dietary supplements of tocotrienols for cardiovascular diseases, as well as others that are based on inflammation.

  19. Interleukin-1 alpha activation and localization in lipopolysaccharide-stimulated human monocytes and macrophages

    DEFF Research Database (Denmark)

    Carlsen, Thomas Gelsing; Kjærsgaard, Pernille

    2015-01-01

    Background: Interleukin-1? (IL-1?) is a proinflammatory cytokine belonging to the IL-1 family. It is synthesized as a 33 kDa precursor peptide that is cleaved by a calpain-like protease to a 16 kDa propiece and a 17 kDa mature IL-1? peptide. In contrast to its close relative, IL-1?, the role of IL- 1? in inflammation is only partly understood. Results: Human macrophages/monocytes, stimulated with lipopolysaccharide (LPS) were analyzed for production and localization of IL-1? by use of a monoclonal antibody (MAb) generated against IL-1? pro piece. We found that IL-1? propiece was detected within the nuclei of the cells 2 hours (hrs) after LPS stimulation and production continued for up to 20 hrs. The MAb was conjugated to fluorescein isothiocyanate (FITC) for use in flow cytometry. Based on the flow cytometric analysis CD68 positive cells were positive for IL-1? in agreement with CD68 being a marker for monocytes. Conclusions: Here, we demonstrate, for the first time, a method to visualize and measure the production of IL-1? in both human monocytes and macrophages.

  20. Antigenic properties of peptidic mimics for epitopes of the lipopolysaccharide from Brucella.

    Science.gov (United States)

    De Bolle, X; Laurent, T; Tibor, A; Godfroid, F; Weynants, V; Letesson, J J; Mertens, P

    1999-11-19

    The lipopolysaccharide (LPS) is up to now the only identified major virulence determinant of Brucella. This bacterium is responsible for brucellosis in animals and for Malta fever in humans. Several monoclonal antibodies (mAbs) directed against various LPS epitopes have been characterized. Two mAbs, named A15-6B3 and B66-2C8, directed against distinct LPS epitopes have been used to select peptides from 11 phage display libraries. The sequences of the selected peptides contain an overrepresentation of either proline or tryptophan residues when selected with either A15-6B3 or B66-2C8 mAbs, respectively. For the best binding peptides, competition with LPS for the binding to the mAb is detected, which suggests that the peptides bind to the paratope of the mAb. The phages selected from the libraries were used to immunise mice, and a weak antibody response directed against LPS has been observed. These data suggest that a subset of the selected peptides are mimotopes of the LPS epitopes. PMID:10556037

  1. Potential use of gallium-doped phosphate-based glass material for periodontitis treatment.

    Science.gov (United States)

    Sahdev, Rohan; Ansari, Tahera I; Higham, Susan M; Valappil, Sabeel P

    2015-07-01

    This study aimed at evaluating the potential effect of gallium-incorporated phosphate-based glasses towards periodontitis-associated bacteria, Porphyromonas gingivalis, and matrix metalloproteinase-13. Periodontitis describes a group of inflammatory diseases of the gingiva and supporting structures of the periodontium. They are initiated by the accumulation of plaque bacteria, such as the putative periodontal pathogen Porphyromonas gingivalis, but the host immune response such as elevated matrix metalloproteinases are the major contributing factor for destruction of periodontal tissues. Antibacterial assays of gallium-incorporated phosphate-based glasses were conducted on Porphyromonas gingivalis ATCC 33277 using disc diffusion assay on fastidious anaerobe agar and liquid broth assay in a modified tryptic soy broth. In vitro study investigated the effect of gallium on purified recombinant human matrix metalloproteinase-13 activity using matrix metalloproteinase assay kit. In vivo biocompatibility of gallium-incorporated phosphate-based glass was evaluated in rats as subcutaneous implants. Antibacterial assay of gallium displayed activity against Porphyromonas gingivalis (inhibition zone of 22?±?0.5?mm compared with 0?mm for control glass, c-PBG). Gallium in the glass contributed to growth inhibitory effect on Porphyromonas gingivalis (up to 1.30 reductions in log?10 values of the viable counts compared with control) in a modified tryptic soy broth. In vitro study showed gallium-incorporated phosphate-based glasses inhibited matrix metalloproteinase activity significantly (p???0.01) compared with c-PBG. Evaluation of in vivo biocompatibility of gallium-incorporated phosphate-based glasses in rats showed a non-toxic and foreign body response after 2 weeks of implantation. The results indicate that gallium ions might act on multiple targets of biological mechanisms underlying periodontal disease. Moreover, gallium-incorporated phosphate-based glasses are biocompatible in a rat model. The findings warrant further investigation and will have important clinical implications in the future treatment and management of periodontitis. PMID:25681404

  2. Effect of probiotics Lactobacillus and Bifidobacterium on gut-derived lipopolysaccharides and inflammatory cytokines: an in vitro study using a human colonic microbiota model.

    Science.gov (United States)

    Rodes, Laetitia; Khan, Afshan; Paul, Arghya; Coussa-Charley, Michael; Marinescu, Daniel; Tomaro-Duchesneau, Catherine; Shao, Wei; Kahouli, Imen; Prakash, Satya

    2013-04-01

    Gut-derived lipopolysaccharides (LPS) are critical to the development and progression of chronic low-grade inflammation and metabolic diseases. In this study, the effects of probiotics Lactobacillus and Bifidobacterium on gut-derived lipopolysaccharide and inflammatory cytokine concentrations were evaluated using a human colonic microbiota model. Lactobacillus reuteri, L. rhamnosus, L. plantarum, Bifidobacterium animalis, B. bifidum, B. longum, and B. longum subsp. infantis were identified from the literature for their anti-inflammatory potential. Each bacterial culture was administered daily to a human colonic microbiota model during 14 days. Colonic lipopolysaccharides, and Gram-positive and negative bacteria were quantified. RAW 264.7 macrophage cells were stimulated with supernatant from the human colonic microbiota model. Concentrations of TNF-alpha, IL-1beta, and IL-4 cytokines were measured. Lipopolysaccharide concentrations were significantly reduced with the administration of B. bifidum (-46.45 +/- 5.65%), L. rhamnosus (-30.40 +/- 5.08%), B. longum (-42.50 +/- 1.28%), and B. longum subsp. infantis (-68.85 +/- 5.32%) (p TNF-alpha concentrations (-69.41 +/- 2.78%; p TNF-alpha and IL-1beta concentrations (p < 0.05). These findings suggest that specific probiotic bacteria, such as B. longum subsp. infantis, might decrease colonic lipopolysaccharide concentrations, which might reduce the proinflammatory tone. This study has noteworthy applications in the field of biotherapeutics for the prevention and/or treatment of inflammatory and metabolic diseases. PMID:23568206

  3. Intra-articular morphine in horses : clinical properties in lipopolysaccharide-induced synovitis

    DEFF Research Database (Denmark)

    Lindegaard, Casper

    2009-01-01

    Optimal smertebehandling af dyr der udsættes for forskellige smertefulde procedurer er yderst vigtigt pga. dyrevelfærdsmæssige årsager såvel som mulighed for forbedret rekonvalescens og forbedret endeligt resultat. Forbedret smertebehandling hos heste kan blandt andet opnås gennem "balanceret smertebehandling". Dette koncept er baseret på at anvende en kombination af flere forskellige smertestillende stoffer som hver især virker på forskellige niveauer af smertesignalets vej til hjernen. Denne kombination af forskellige typer analgetika giver en additiv, og i nogle tilfælde også synergistisk effekt. Dette medfører at dosen af hvert enkelt analgetikum kan reduceres, hvilket fører til at mængden af bivirkninger reduceres. Intraartikulært (IA) administreret morfin har vist sig at virke smertestillende i op til 24 timer efter artroskopisk kirurgi hos mennesker. Derudover er det vist at IA morfin også besidder betydelige antiinflammatoriske egenskaber i mennesker og forsøgsdyr. Intraartikulært administreret morfin anvendes derfor som en del af en balanceret smertebehandlingsprotokol efter artroskopisk kirurgi, eller ved andre smertefulde ledlidelser hos mennesker. For nylig har opdagelsen af opioid receptorer i ledkapslen hos heste ført til en forventning om lignende egenskaber af IA morfin hos heste. På trods af at der indtil videre ikke er udført nogen forskning på området, er intraartikulær injektion af morfin blevet fast procedure efter artroskopisk kirurgi på adskillige veterinær-universiteter i Europa og USA. Formålet med denne afhandling var at undersøge den smertestillende og antiinflammatoriske effekt, samt farmakokinetikken af IA morfin hos heste med ledinflammation. For at undersøge dette blev der udført et forsøg med lipopolysaccharid-induceret synovitis hos 8 heste.  Studiet blev udført som et randomiseret, blændet, double dummy forsøg med sekventielt cross-over design. Alle heste modtog 2 behandlinger: "IA morfin" som bestod af morfin 0,05 mg/kg IA + fysiologisk saltvand IV og "IV morfin" som bestod af morfin 0,05 mg/kg IV + fysiologisk saltvand IA. De to behandlinger blev givet i tilfældig rækkefølge med en 3 ugers restitutions periode ind imellem. Før administration af forsøgsbehandlingerne blev der ved IA injektion af lipopolysaccharid (LPS), induceret synovitis i et radiocarpalled. I hver af de to 7-dages forsøgsperioder blev forskellige lokale og systemiske inflammations- og smerteparametre målt. Derudover blev der fortaget farmakologiske undersøgelser på blod og ledvæske. Smerte blev vurderet vha. halthed samt smerte vurderet på subjektiv smerteskala (Visuel analog skala, VAS) og en adfærdsbaseret smerteskala (Composite Measure Pain Scale, CMPS). Hos alle heste inducerede de intraartikulære LPS injektioner led-inflammation (synovitis) og deraf følgende smerte og halthed. Morfin injiceret IA havde en signifikant smertestillende effekt, målt ved signifikant reduceret halthed, lavere forbrug af rescue analgesi samt lavere smertescorer. Den anvendte CMPS smerteskala viste god overensstemmelse mellemobservatørerne og det forslås at den kan anvendes til vurdering af ortopædisk smerte hos heste. Derudover sås en signifikant antiinflammatorisk effekt, som kom til udtryk ved reduceret ledhævelse, reduceret indhold af SAA og protein i ledvæsken samt reduceret SAA i blodet. Den antiinflammatoriske effekt syntes mest udtalt sent i den inflammatoriske proces. Ved analyser for morfin og de to metabolitter morfin-3- og morfin-6-glucuronid (hhv. M3G og M6G) fandtes morfin i ledvæsken fra alle 8 heste 24 timer efter den intraartikulære injektion. Samtidig var serum koncentrationerne af morfin og den aktive metabolit M6G under klinisk relevante niveauer fra 2 timer efter behandlingen og frem. Derudover blev det vist at farmakokinetikken af IA morfin hos heste er sammenlignelig med det som er observeret hos mennesker. Da behandling med morfin IA resulterede i mindre smerte og inflammation end behandling med samme dosis morfin givet systemisk indikerer dette, sammen me

  4. Occurrence of an unusual hopanoid-containing lipid A among lipopolysaccharides from Bradyrhizobium species.

    Science.gov (United States)

    Komaniecka, Iwona; Choma, Adam; Mazur, Andrzej; Duda, Katarzyna A; Lindner, Buko; Schwudke, Dominik; Holst, Otto

    2014-12-19

    The chemical structures of the unusual hopanoid-containing lipid A samples of the lipopolysaccharides (LPS) from three strains of Bradyrhizobium (slow-growing rhizobia) have been established. They differed considerably from other Gram-negative bacteria in regards to the backbone structure, the number of ester-linked long chain hydroxylated fatty acids, as well as the presence of a tertiary residue that consisted of at least one molecule of carboxyl-bacteriohopanediol or its 2-methyl derivative. The structural details of this type of lipid A were established using one- and two-dimensional NMR spectroscopy, chemical composition analyses, and mass spectrometry techniques (electrospray ionization Fourier-transform ion cyclotron resonance mass spectrometry and MALDI-TOF-MS). In these lipid A samples the glucosamine disaccharide characteristic for enterobacterial lipid A was replaced by a 2,3-diamino-2,3-dideoxy-d-glucopyranosyl-(GlcpN3N) disaccharide, deprived of phosphate residues, and substituted by an ?-d-Manp-(1?6)-?-d-Manp disaccharide substituting C-4' of the non-reducing (distal) GlcpN3N, and one residue of galacturonic acid (d-GalpA) ?-(1?1)-linked to the reducing (proximal) amino sugar residue. Amide-linked 12:0(3-OH) and 14:0(3-OH) were identified. Some hydroxy groups of these fatty acids were further esterified by long (?-1)-hydroxylated fatty acids comprising 26-34 carbon atoms. As confirmed by mass spectrometry techniques, these long chain fatty acids could form two or three acyloxyacyl residues. The triterpenoid derivatives were identified as 34-carboxyl-bacteriohopane-32,33-diol and 34-carboxyl-2?-methyl-bacteriohopane-32,33-diol and were covalently linked to the (?-1)-hydroxy group of very long chain fatty acid in bradyrhizobial lipid A. Bradyrhizobium japonicum possessed lipid A species with two hopanoid residues. PMID:25371196

  5. Spontaneously hypertensive rats display reduced microglial activation in response to ischemic stroke and lipopolysaccharide

    Directory of Open Access Journals (Sweden)

    De Geyter Deborah

    2012-05-01

    Full Text Available Abstract Background For successful translation to clinical stroke studies, the Stroke Therapy Academic Industry Round Table criteria have been proposed. Two important criteria are testing of therapeutic interventions in conscious animals and the presence of a co-morbidity factor. We chose to work with hypertensive rats since hypertension is an important modifiable risk factor for stroke and influences the clinical outcome. We aimed to compare the susceptibility to ischemia in hypertensive rats with those in normotensive controls in a rat model for induction of ischemic stroke in conscious animals. Methods The vasoconstrictor endothelin-1 was stereotactically applied in the vicinity of the middle cerebral artery of control Wistar Kyoto rats (WKYRs and Spontaneously Hypertensive rats (SHRs to induce a transient decrease in striatal blood flow, which was measured by the Laser Doppler technique. Infarct size was assessed histologically by Cresyl Violet staining. Sensory-motor functions were measured at several time points using the Neurological Deficit Score. Activation of microglia and astrocytes in the striatum and cortex was investigated by immunohistochemistry using antibodies against CD68/Iba-1 and glial fibrillary acidic protein. Results and conclusions The SHRs showed significantly larger infarct volumes and more pronounced sensory-motor deficits, compared to the WKYRs at 24?h after the insult. However, both differences disappeared between 24 and 72?h. In SHRs, microglia were less susceptible to activation by lipopolysaccharide and there was a reduced microglial activation after induction of ischemic stroke. These quantitative and qualitative differences may be relevant for studying the efficacy of new treatments for stroke in accordance to the Stroke Therapy Academic Industry Round Table criteria.

  6. Marginal zinc deficiency increased the susceptibility to acute lipopolysaccharide-induced liver injury in rats.

    Science.gov (United States)

    Shea-Budgell, Melissa; Dojka, Marie; Nimmo, Michael; Lee, Diana; Xu, Zhaoming

    2006-05-01

    Lipopolysaccharide (LPS) triggers a global activation of inflammatory responses leading to liver injury in humans. Zinc pretreatment has been shown to prevent LPS-induced hepatic necrosis. In North America, suboptimal zinc status is more common than once realized. However, the effect of inadequate zinc nutrition on the host's susceptibility to LPS-induced liver injury is not known. The objective of this study was to determine whether marginal zinc deficiency would render rats more susceptible to LPS-induced liver injury. Weanling Sprague-Dawley rats were assigned to one of three dietary treatment groups: marginally low zinc ad libitum (Z3; 3 mg zinc/kg diet), adequate zinc ad libitum (Z30; 30 mg zinc/kg diet), or adequate zinc pair-fed (Z30P) group. After 6 weeks, each dietary treatment group was further divided into LPS-control (saline) groups (C-Z3, C-Z30P, C-Z30) and LPS-treatment (1 mg/kg body weight, intraperitoneal, 8 hrs) groups (LPS-Z3, LPS-Z30P, LPS-Z30). LPS reduced the serum zinc concentration and increased the liver zinc concentration regardless of dietary zinc intake. Serum alanine aminotransferase level was higher in the LPS-Z3 rats than in the LPS-Z30P and LPS-Z30 rats. LPS also induced hepatocyte necrosis and neutrophil infiltration into the liver sinusoids. This LPS-induced liver damage was more severe in the LPS-Z3 rats than in the LPS-Z30P and LPS-Z30 rats. Together these findings have demonstrated that marginal zinc deficiency increased the susceptibility to LPS-induced liver injury in rats. These results indicate that patients with sepsis who have suboptimal zinc nutrition status may be at higher risk of developing greater liver damage. PMID:16636303

  7. Inhaled nitric oxide reduces tyrosine nitration after lipopolysaccharide instillation into lungs of rats.

    Science.gov (United States)

    Honda, K; Kobayashi, H; Hataishi, R; Hirano, S; Fukuyama, N; Nakazawa, H; Tomita, T

    1999-08-01

    Nitric oxide (NO) may either protect against or contribute to inflammatory lung injury. In this study we investigated whether inhalation of 20 ppm NO alters tyrosine nitration, and we assessed the degree of lung inflammation and edema in rats after lipopolysaccharide (LPS) instillation. The amount of nitrotyrosine relative to the total amount of tyrosine was measured in lung homogenates, and lung tissue sections were stained for nitrotyrosine and aminotyrosine (a reduced form of nitrotyrosine). Leukocytes in bronchoalveolar lavage fluid (BALF) were counted, and myeloperoxidase activity was measured in lung homogenate. Lung edema and inflammatory cell accumulation in lung tissue were estimated by extravascular lung water weight (EVLW) and extravascular dry lung weight (EVDW), respectively. LPS instillation caused increases in nitrotyrosine concentration and immunohistochemical staining of nitrotyrosine and aminotyrosine in the lungs. LPS instillation increased the BALF leukocyte count, myeloperoxidase activity in lung tissue, and both EVLW and EVDW. Inhalational exposure to 20 ppm NO induced nitrotyrosine and aminotyrosine formation only in bronchial epithelial cell surface of the lungs not instilled with LPS. NO inhalation reduced the increases in nitrotyrosine and aminotyrosine in LPS-instilled lung tissue as well as the leukocyte count in BALF and myeloperoxidase activity in lung tissue, but it did not significantly change EVLW or EVDW. Leukocyte depletion in LPS-instilled rats reduced interstitial inflammatory cells, which were stained with nitrotyrosine and aminotyrosine, and attenuated the nitrotyrosine staining of alveolar capillaries. These results suggest that inhalation of 20 ppm NO reduces leukocyte accumulation in the lungs and inhibits tyrosine nitration caused by LPS instillation. PMID:10430746

  8. Salvianolic acid A suppress lipopolysaccharide-induced NF-?B signaling pathway by targeting IKK?.

    Science.gov (United States)

    Oh, Kwang-Seok; Oh, Byung Koo; Mun, Jihye; Seo, Ho Won; Lee, Byung Ho

    2011-11-01

    Salvianolic acid A, an active compound present in Salvia miltiorrhiza, is a phenolic carboxylic acid derivative, ((2R)-3-(3,4-Dihydroxyphenyl)-2-[(E)-3-[2-[(E)-2-(3,4-dihydroxyphenyl) ethenyl]-3,4-dihydroxyphenyl] prop-2-enoyl]oxypropanoic acid). The present study was performed to investigate the underlying mechanisms of anti-inflammatory effects with salvianolic acid A, specially focused on nuclear factor ?B (NF-?B) signaling pathway by targeting the I?B kinase ? (IKK?). The effect of salvianolic acid A for IKK? activity was analyzed using an immobilized metal affinity for phosphochemicals (IMAP)-based time-resolved fluorescence resonance energy transfer (TR-FRET) assay. The underlying mechanisms of salvianolic acid A were examined using lipopolysaccharide (LPS)-stimulated RAW264.7 cells. IKK? studies based on IMAP-TR-FRET showed that salvianolic acid A possesses a potent IKK? inhibitory activity with Ki value of 3.63 ?M in an ATP-noncompetitive manner. Pretreatment with salvianolic acid A (10, 30 ?M) decreased LPS-induced expression of iNOS and COX-2, thereby inhibiting production of nitric oxide and prostaglandin E(2), respectively. In addition, salvianolic acid A (10, 30 ?M) also attenuated the LPS-induced I?B? phosphorylation and degradation, and NF-?B translocation. These results suggest that salvianolic acid A modulates NF-?B-dependent inflammatory pathways through IKK? inhibition and these anti-inflammatory effects will aid in understanding the pharmacology and mode of action of salvianolic acid A. PMID:21839184

  9. Influence of lipopolysaccharide on the uterine contraction inhibitory effects of tocolytic agents in pregnant mice.

    Science.gov (United States)

    Sugawara, Noboru; Okawa, Toshiaki; Takahashi, Hidenori; Sato, Akira; Vedernikov, Yuri P; Saade, George R; Garfield, Robert E

    2007-10-01

    The study was undertaken to investigate the influence of lipopolysaccharide (LPS) on the uterine contraction inhibitory effects of tocolytic agents such as ritodrine, magnesium, and diethylamine/nitric oxide (DEA/NO), and on prostaglandin (PG) E2 and nitric oxide (NO) metabolite (NOx) production in pregnant mice at midgestation. Pregnant C57BL mice on day 14 of gestation were sacrificed 6 hours after intraperitoneal injection of LPS (400 mug/kg) or vehicle. Uterine rings were equilibrated in Krebs-Henseleit solution (37 degrees C) bubbled with 20% O (2) and 5% CO (2) (pH ~7.4) for sampling and isometric tension recording. The concentration levels of PGE2 and NOx in the solution were determined. Changes of spontaneous contractile activity in response to cumulative concentrations of ritodrine, magnesium, and the NO donor, DEA/NO, from the baseline were determined. Integral contractile activity over 10 minutes at each concentration was calculated and expressed as percentage change from basal activity. Statistical analysis was performed using one-way analysis of variance (ANOVA) followed by the Dunnett test (significance: P NOx (from 66.8 +/- 6.7 pg/g tissue to 147.0 +/- 29.0, from 51.0 +/- 5.4 pmol/2 muL/g tissue to 98.0 +/- 16.2, respectively), P concentration dependently in uterine rings from both LPS-treated and -untreated animals. Treatment with LPS significantly attenuated the maximal inhibition induced by DEA/NO in uterine rings from pregnant mice. LPS significantly suppressed the uterine contraction inhibitory effects of ritodrine at 10 (-8) M concentrations and of magnesium at 4.2 mmol concentration ( P NOx, which cause increased spontaneous contractions of the uterine myometrium. Therefore, when uterine contractions are not controlled by tocolytics in pregnant patients with preterm labor associated with inflammation, labor induction or pregnancy termination may become significant options in clinical practice. PMID:17907072

  10. Hesperidin ameliorates lipopolysaccharide-induced acute lung injury in mice by inhibiting HMGB1 release.

    Science.gov (United States)

    Liu, Xin-xin; Yu, Dan-dan; Chen, Mao-jian; Sun, Ting; Li, Gang; Huang, Wen-jian; Nie, Hao; Wang, Chao; Zhang, Yan-xiang; Gong, Quan; Ren, Bo-xu

    2015-04-01

    Hesperidin (HDN), a flavanone glycoside, possesses anti-inflammatory properties and has been suggested to be able to modulate the lipopolysaccharide (LPS)-induced acute lung injury (ALI). High-mobility group box 1 (HMGB1) serves as an inflammatory cytokine when released extracellularly and is involved in the pathogenesis of diverse inflammatory disorders. The current study aimed to investigate the involvement of HMGB1 in HDN-induced immunoregulation of ALI. ALI in male BALB/c mice was induced by intranasal administration of LPS (0.5mg/kg). HDN (500mg/kg) was administered intragastrically 10days prior to LPS exposure. HDN significantly protected animals from LPS-induced ALI as evidenced by decreased elevation of the lung wet to dry weight ratio, total cells, neutrophils, macrophages, and myeloperoxidase (MPO) activity, associated with reduced lung histological damage. In the meantime, HDN pretreatment markedly inhibited the production of pro-inflammatory cytokines and chemokine, including tumor necrosis factor-? (TNF-?), interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1). Furthermore, HDN pretreatment dramatically inhibited the infiltration of macrophages and suppressed the expression and release of HMGB1 in vivo and in vitro. In addition, intranasal application of exogenous HMGB1 could result in lung injury which was also alleviated by HDN administration. These results suggest that HDN pretreatment protects mice from LPS-induced ALI via inhibiting the production of TNF-? and IL-6. Moreover, we found that HDN could inhibit the expression and release of HMGB1 via suppressing the infiltration of macrophages and production of MCP-1. PMID:25724384

  11. Milk thistle extract and silymarin inhibit lipopolysaccharide induced lamellar separation of hoof explants in vitro.

    Science.gov (United States)

    Reisinger, Nicole; Schaumberger, Simone; Nagl, Veronika; Hessenberger, Sabine; Schatzmayr, Gerd

    2014-10-01

    The pathogenesis of laminitis is not completely identified and the role of endotoxins (lipopolysaccharides, LPS) in this process remains unclear. Phytogenic substances, like milk thistle (MT) and silymarin, are known for their anti-inflammatory and antioxidant properties and might therefore have the potential to counteract endotoxin induced effects on the hoof lamellar tissue. The aim of our study was to investigate the influence of endotoxins on lamellar tissue integrity and to test if MT and silymarin are capable of inhibiting LPS-induced effects in an in vitro/ex vivo model. In preliminary tests, LPS neutralization efficiency of these phytogenics was determined in an in vitro neutralization assay. Furthermore, tissue explants gained from hooves of slaughter horses were tested for lamellar separation after incubation with different concentrations of LPS. By combined incubation of explants with LPS and either Polymyxin B (PMB; positive control), MT or silymarin, the influence of these substances on LPS-induced effects was assessed. In the in vitro neutralization assay, MT and silymarin reduced LPS concentrations by 64% and 75%, respectively, in comparison PMB reduced 98% of the LPS concentration. In hoof explants, LPS led to a concentration dependent separation. Accordantly, separation force was significantly decreased by 10 µg/mL LPS. PMB, MT and silymarin could significantly improve tissue integrity of explants incubated with 10 µg/mL LPS. This study showed that LPS had a negative influence on the structure of hoof explants in vitro. MT and silymarin reduced endotoxin activity and inhibited LPS-induced effects on the lamellar tissue. Hence, MT and silymarin might be used to support the prevention of laminitis and should be further evaluated for this application. PMID:25290524

  12. Milk Thistle Extract and Silymarin Inhibit Lipopolysaccharide Induced Lamellar Separation of Hoof Explants in Vitro

    Directory of Open Access Journals (Sweden)

    Nicole Reisinger

    2014-10-01

    Full Text Available The pathogenesis of laminitis is not completely identified and the role of endotoxins (lipopolysaccharides, LPS in this process remains unclear. Phytogenic substances, like milk thistle (MT and silymarin, are known for their anti-inflammatory and antioxidant properties and might therefore have the potential to counteract endotoxin induced effects on the hoof lamellar tissue. The aim of our study was to investigate the influence of endotoxins on lamellar tissue integrity and to test if MT and silymarin are capable of inhibiting LPS-induced effects in an in vitro/ex vivo model. In preliminary tests, LPS neutralization efficiency of these phytogenics was determined in an in vitro neutralization assay. Furthermore, tissue explants gained from hooves of slaughter horses were tested for lamellar separation after incubation with different concentrations of LPS. By combined incubation of explants with LPS and either Polymyxin B (PMB; positive control, MT or silymarin, the influence of these substances on LPS-induced effects was assessed. In the in vitro neutralization assay, MT and silymarin reduced LPS concentrations by 64% and 75%, respectively, in comparison PMB reduced 98% of the LPS concentration. In hoof explants, LPS led to a concentration dependent separation. Accordantly, separation force was significantly decreased by 10 µg/mL LPS. PMB, MT and silymarin could significantly improve tissue integrity of explants incubated with 10 µg/mL LPS. This study showed that LPS had a negative influence on the structure of hoof explants in vitro. MT and silymarin reduced endotoxin activity and inhibited LPS-induced effects on the lamellar tissue. Hence, MT and silymarin might be used to support the prevention of laminitis and should be further evaluated for this application.

  13. Lipopolysaccharide Does Not Alter Small Airway Reactivity in Mouse Lung Slices

    Science.gov (United States)

    Donovan, Chantal; Royce, Simon G.; Vlahos, Ross; Bourke, Jane E.

    2015-01-01

    The bacterial endotoxin, lipopolysaccharide (LPS) has been associated with occupational airway diseases with asthma-like symptoms and in acute exacerbations of COPD. The direct and indirect effects of LPS on small airway reactivity have not been fully elucidated. We tested the hypothesis that both in vitro and in vivo LPS treatment would increase contraction and impair relaxation of mouse small airways. Lung slices were prepared from naïve Balb/C mice and cultured in the absence or presence of LPS (10 ?g/ml) for up to 48 h for measurement of TNF? levels in conditioned media. Alternatively, mice were challenged with PBS or LPS in vivo once a day for 4 days for preparation of lung slices or for harvest of lungs for Q-PCR analysis of gene expression of pro-inflammatory cytokines and receptors involved in airway contraction. Reactivity of small airways to contractile agonists, methacholine and serotonin, and bronchodilator agents, salbutamol, isoprenaline and rosiglitazone, were assessed using phase-contrast microscopy. In vitro LPS treatment of slices increased TNF? release 6-fold but did not alter contraction or relaxation to any agonists tested. In vivo LPS treatment increased lung gene expression of TNF?, IL-1? and ryanodine receptor isoform 2 more than 5-fold. However there were no changes in reactivity in lung slices from these mice, even when also incubated with LPS ex vivo. Despite evidence of LPS-induced inflammation, neither airway hyperresponsiveness or impaired dilator reactivity were evident. The increase in ryanodine receptor isoform 2, known to regulate calcium signaling in vascular smooth muscle, warrants investigation. Since LPS failed to elicit changes in small airway reactivity in mouse lung slices following in vitro or in vivo treatment, alternative approaches are required to define the potential contribution of this endotoxin to altered small airway reactivity in human lung diseases. PMID:25822969

  14. Exposure of periodontal ligament progenitor cells to lipopolysaccharide from Escherichia coli changes osteoblast differentiation pattern

    Scientific Electronic Library Online (English)

    Mayra Laino, ALBIERO; Bruna Rabelo, AMORIM; Luciane, MARTINS; Márcio Zaffalon, CASATI; Enilson Antonio, SALLUM; Francisco Humberto, NOCITI JR; Karina Gonzales, SILVÉRIO.

    2015-04-01

    Full Text Available Periodontal ligament mesenchymal stem cells (PDLMSCs) are an important alternative source of adult stem cells and may be applied for periodontal tissue regeneration, neuroregenerative medicine, and heart valve tissue engineering. However, little is known about the impact of bacterial toxins on the b [...] iological properties of PDLSMSCs, including self-renewal, differentiation, and synthesis of extracellular matrix. Objective : This study investigated whether proliferation, expression of pro-inflammatory cytokines, and osteogenic differentiation of CD105-enriched PDL progenitor cell populations (PDL-CD105+ cells) would be affected by exposure to bacterial lipopolysaccharide from Escherichia coli (EcLPS). Material and Methods : Toll-like receptor 4 (TLR4) expression was assessed in PDL-CD105+ cells by the immunostaining technique and confirmed using Western blotting assay. Afterwards, these cells were exposed to EcLPS, and the following assays were carried out: (i) cell viability using MTS; (ii) expression of the interleukin-1 beta (IL-1?), interleukin-6 (IL-6), interleukin-8 (IL-8), and tumor necrosis factor alpha (TNF-?) genes; (iii) osteoblast differentiation assessed by mineralization in vitro, and by mRNA levels of run-related transcription factor-2 (RUNX2), alkaline phosphatase (ALP) and osteocalcin (OCN) determined by quantitative PCR. Results : PDL-CD105+ cells were identified as positive for TLR4. EcLPS did not affect cell viability, but induced a significant increase of transcripts for IL-6 and IL-8. Under osteogenic condition, PDL-CD105+ cells exposed to EcLPS presented an increase of mineralized matrix deposition and higher RUNX2 and ALP mRNA levels when compared to the control group. Conclusions : These results provide evidence that CD105-enriched PDL progenitor cells are able to adapt to continuous Escherichia coli endotoxin challenge, leading to an upregulation of osteogenic activities.

  15. Microarray Analysis of Human Vascular Smooth Muscle Cell Responses to Bacterial Lipopolysaccharide

    Directory of Open Access Journals (Sweden)

    Joe Minta

    2007-01-01

    Full Text Available Accumulating evidence suggest a causal role of bacterial and viral infections in atherogenesis. Bacterial lipopolysaccharide (LPS has been shown to stimulate resting vascular smooth muscle cells (SMC with the production of inflammatory cytokines and modulation of quiescent cells to the proliferative and synthetic phenotype. To comprehensively identify biologically important genes associated with LPS-induced SMC phenotype modulation, we compared the transcriptomes of quiescent human coronary artery SMC and cells treated with LPS for 4 and 22 h. The SMCs responded robustly to LPS treatment by the differential regulation of several genes involved in chromatin remodeling, transcription regulation, translation, signal transduction, metabolism, host defense, cell proliferation, apoptosis, matrix formation, adhesion and motility and suggest that the induction of clusters of genes involved in cell proliferation, migration and ECM production may be the main force that drives the LPS-induced phenotypic modulation of SMC rather than the differential expression of a single gene or a few genes. An interesting observation was the early and dramatic induction of four tightly clustered interferon-induced genes with tetratricopeptide repeats (IFIT1, 2, 4, 5. siRNA knock-down of IFIT1 in SMC was found to be associated with a remarkable up-regulation of TP53, CDKN1A and FOS, suggesting that IFIT1 may play a role in cell proliferation. Our data provide a comprehensive list of genes involved in LPS biology and underscore the important role of LPS in SMC activation and phenotype modulation which is a pivotal event in the onset of atherogenesis.

  16. Systemic administration of lipopolysaccharide in laying hens stimulates antimicrobial properties of egg white against Staphylococcus aureus.

    Science.gov (United States)

    Bedrani, Larbi; Helloin, Emmanuelle; Guyot, Nicolas; Nys, Yves

    2013-04-15

    The natural protective system of eggs relies on egg yolk immunoglobulins and on antimicrobial proteins/peptides mainly concentrated in the egg white. There is much evidence concerning the specific stimulation of immunoglobulins by antigens but to date, the influence of the hen milieu on the regulation of the egg innate molecular immunity has not been established. To explore the hypothesis of modulation in egg antimicrobial molecules, laying hens were immune-challenged with intravenous injections of Salmonella enterica Enteritidis lipopolysaccharide (LPS) at 24 h intervals. Eggs of the control and LPS groups were collected over a period of 21 days following the first LPS injection and the egg white activities against Staphylococcus aureus and Escherichia coli were assessed. The increase in egg white anti-S. aureus activity reached 20.9% and 23.4% (plysozyme and proteases inhibiting (anti-trypsin and anti-chymotrypsin) activities and the pH variations of egg whites. We recorded no significant variations between the two experimental groups for these potential modulating factors. Finally, using RT-qPCR we studied the expression of several genes coding for antimicrobial proteins and peptides involved in the immune response in the infundibulum and the magnum, Out of the 11 genes, only TLR4 in the magnum and ovocalyxin-36 in infundibulum were over-expressed respectively 24h and 8 days after the first LPS injection. The other candidate genes showed similar or down regulated expression in the LPS group as compared to the control especially during the first 24h. Our results suggest that the hen enhances the albumen antimicrobial activity of its eggs when exposed to immune stimulations or infections. This could be an attempt to preventively reinforce the protection of the embryo with nonspecific antimicrobial agents in addition to the specific antibodies exported to the egg. The origin of this stimulation of egg molecular immunity remains to be characterized amongst the numerous novel egg proteins recently identified. PMID:23351641

  17. Clinical and veterinary isolates of Salmonella enterica serovar Enteritidis defective in lipopolysaccharide O-chain polymerization

    Energy Technology Data Exchange (ETDEWEB)

    Guard-Petter, J.; Parker, C.T. [Agricultural Research Service, Athens, GA (United States). Southeast Poultry Research Lab.; Asokan, K.; Carlson, R.W. [Univ. of Georgia, Athens, GA (United States). Complex Carbohydrate Research Center

    1999-05-01

    Twelve human and chicken isolates of Salmonella enterica serovar Enteritidis belonging to phage types 4, 8, 13a, and 23 were characterized for variability in lipopolysaccharide (LPS) composition. Isolates were differentiated into two groups, i.e., those that lacked immunoreactive O-chain, termed rough isolates, and those that had immunoreactive O-chain, termed smooth isolates. Isolates within these groups could be further differentiated by LPS compositional differences as detected by gel electrophoresis and gas liquid chromatography of samples extracted with water, which yielded significantly more LPS in comparison to phenol-chloroform extraction. The rough isolates were of two types, the O-antigen synthesis mutants and the O-antigen polymerization (wzy) mutants. Smooth isolates were also of two types, one producing low-molecular-weight (LMW) LPS and the other producing high-molecular-weight (HMW) LPS. To determine the genetic basis for the O-chain variability of the smooth isolates, the authors analyzed the effects of a null mutation in the O-chain length determinant gene, wzz (cld) of serovar Typhimurium. This mutation results in a loss of HMW LPS; however, the LMW LPS of this mutant was longer and more glucosylated than that from clinical isolates of serovar Enteritidis. Cluster analysis of these data and of those from two previously characterized isogenic strains of serovar Enteritidis that had different virulence attributes indicated that glucosylation of HMW LPS (via oafR function) is variable and results in two types of HMW structures, one that is highly glucosylated and one that is minimally glucosylated. These results strongly indicate that naturally occurring variability in wzy, wzz, and oafR function can be used to subtype isolates of serovar Enteritidis during epidemiological investigations.

  18. Effects of Acarbose Addition on Ruminal Bacterial Microbiota, Lipopolysaccharide Levels and Fermentation Characteristics In vitro.

    Science.gov (United States)

    Yin, Yu-Yang; Liu, Yu-Jie; Zhu, Wei-Yun; Mao, Sheng-Yong

    2014-12-01

    This study investigated the effects of acarbose addition on changes in ruminal fermentation characteristics and the composition of the ruminal bacterial community in vitro using batch cultures. Rumen fluid was collected from the rumens of three cannulated Holstein cattle fed forage ad libitum that was supplemented with 6 kg of concentrate. The batch cultures consisted of 8 mL of strained rumen fluid in 40 mL of an anaerobic buffer containing 0.49 g of corn grain, 0.21 g of soybean meal, 0.15 g of alfalfa and 0.15g of Leymus chinensis. Acarbose was added to incubation bottles to achieve final concentrations of 0.1, 0.2, and 0.4 mg/mL. After incubation for 24 h, the addition of acarbose linearly decreased (ptotal gas production and the concentrations of acetate, propionate, butyrate, total volatile fatty acids, lactate and lipopolysaccharide (LPS). It also linearly increased (pnitrogen and the pH value compared with the control. Pyrosequencing of the 16S rRNA gene showed that the addition of acarbose decreased (p<0.05) the proportion of Firmicutes and Proteobacteria and increased (p<0.05) the percentage of Bacteroidetes, Fibrobacteres, and Synergistetes compared with the control. A principal coordinates analysis plot based on unweighted UniFrac values and molecular variance analysis revealed that the structure of the ruminal bacterial communities in the control was different to that of the ruminal microbiota in the acarbose group. In conclusion, acarbose addition can affect the composition of the ruminal microbial community and may be potentially useful for preventing the occurrence of ruminal acidosis and the accumulation of LPS in the rumen. PMID:25358366

  19. Activation of decidual invariant natural killer T cells promotes lipopolysaccharide-induced preterm birth.

    Science.gov (United States)

    Li, Liping; Yang, Jing; Jiang, Yao; Tu, Jiaoqin; Schust, Danny J

    2015-04-01

    Invariant natural killer T (iNKT) cells are crucial for host defense against a variety of microbial pathogens, but the underlying mechanisms of iNKT cells activation by microbes are not fully explained. In this study, we investigated the molecular mechanisms of iNKT cell activation in lipopolysaccharide (LPS)-stimulated preterm birth using an adoptive transfer system and diverse neutralizing antibodies (Abs) and inhibitors. We found that adoptive transfer of decidual iNKT cells to LPS-stimulated iNKT cell deficient J?18(-/-) mice that lack invariant V?14J?281T cell receptor (TCR) expression significantly decreased the time to delivery and increased the percentage of decidual iNKT cells. Neutralizing Abs against Toll-like receptor 4 (TLR-4), CD1d, interleukin (IL)-12 and IL-18, and inhibitors blocking the activation of nuclear factor ?B (NF-?B), mitogen-activated protein kinase (MAPK) p38 and extracellular signal-regulated kinase (ERK) significantly reduced in vivo percentages of decidual iNKT cells, their intracellular interferon (IFN)-? production and surface CD69 expression. In vitro, in the presence of the same Abs and inhibitors used as in vivo, decidual iNKT cells co-cultured with LPS-pulsed dendritic cells (DCs) showed significantly decreased extracellular and intracellular IFN-? secretion and surface CD69 expression. Our data demonstrate that the activation of decidual iNKT cells plays an important role in inflammation-induced preterm birth. Activation of decidual iNKT cells also requires TLR4-mediated NF-?B, MAPK p38 and ERK pathways, the proinflammatory cytokines IL-12 and IL-18, and endogenous glycolipid antigens presented by CD1d. PMID:25589517

  20. Lipopolysaccharide promotes lipid accumulation in human adventitial fibroblasts via TLR4-NF-?B pathway

    Directory of Open Access Journals (Sweden)

    Wang Jun

    2012-10-01

    Full Text Available Abstract Background Atherosclerosis is a chronic degenerative disease of the arteries and is thought to be one of the most common causes of death globally. In recent years, the functions of adventitial fibroblasts in the development of atherosclerosis and tissue repair have gained increased interests. LPS can increase the morbidity and mortality of atherosclerosis-associated cardiovascular disease. Although LPS increases neointimal via TLR4 activation has been reported, how LPS augments atherogenesis through acting on adventitial fibroblasts is still unknown. Here we explored lipid deposition within adventitial fibroblasts mediated by lipopolysaccharide (LPS to imitate inflammatory conditions. Results In our study, LPS enhanced lipid deposition by the up-regulated expression of adipose differentiation-related protein (ADRP as the silencing of ADRP abrogated lipid deposition in LPS-activated adventitial fibroblasts. In addition, pre-treatment with anti-Toll-like receptor 4 (TLR4 antibody diminished the LPS-induced lipid deposition and ADRP expression. Moreover, LPS induced translocation of nuclear factor-?B (NF-?B, which could markedly up-regulate lipid deposition as pre-treatment with the NF-?B inhibitor, PDTC, significantly reduced lipid droplets. In addition, the lowering lipid accumulation was accompanied with the decreased ADRP expression. Furthermore, LPS-induced adventitial fibroblasts secreted more monocyte chemoattractant protein (MCP-1, compared with transforming growth factor-?1 (TGF-?1. Conclusions Taken together, these results suggest that LPS promotes lipid accumulation via the up-regulation of ADRP expression through TLR4 activated downstream of NF-?B in adventitial fibroblasts. Increased levels of MCP-1 released from LPS-activated adventitial fibroblasts and lipid accumulation may accelerate monocytes recruitment and lipid-laden macrophage foam cells formation. Here, our study provides a new explanation as to how bacterial infection contributes to the pathological process of atherosclerosis.

  1. Experimental optic neuritis induced by the microinjection of lipopolysaccharide into the optic nerve.

    Science.gov (United States)

    Aranda, Marcos L; Dorfman, Damián; Sande, Pablo H; Rosenstein, Ruth E

    2015-04-01

    Optic neuritis (ON) is a condition involving primary inflammation, demyelination, and axonal injury in the optic nerve which leads to retinal ganglion cell (RGC) loss, and visual dysfunction. We investigated the ability of a single microinjection of bacterial lipopolysaccharide (LPS) directly into the optic nerve to induce functional and structural alterations compatible with ON. For this purpose, optic nerves from male Wistar rats remained intact or were injected with vehicle or LPS. The effect of LPS was evaluated at several time points post-injection in terms of: i) visual pathway and retinal function (visual evoked potentials (VEPs) and electroretinograms, (ERGs), respectively), ii) anterograde transport from the retina to its projection areas, iii) consensual pupil light reflex (PLR), iv) optic nerve histology, v) microglia/macrophage reactivity (by Iba-1- and ED1-immunostaining), vi) astrocyte reactivity (by glial fibrillary acid protein-immunostaining), vii) axon number (by toluidine blue staining), vii) demyelination (by myelin basic protein immunoreactivity and luxol fast blue staining), viii) optic nerve ultrastructure, and ix) RGC number (by Brn3a immunoreactivity). LPS induced a significant and persistent decrease in VEP amplitude and PLR, without changes in the ERG. In addition, LPS induced a deficit in anterograde transport, and an early inflammatory response consisting in an increased cellularity, and Iba-1 and ED1-immunoreactivity in the optic nerve, which were followed by changes in axonal density, astrocytosis, demyelination, and axon and RGC loss. These results suggest that the microinjection of LPS into the optic nerve may serve as a new experimental model of primary ON. PMID:25687552

  2. Multiple mechanisms involved in diabetes protection by lipopolysaccharide in non-obese diabetic mice.

    Science.gov (United States)

    Wang, Jun; Cao, Hui; Wang, Hongjie; Yin, Guoxiao; Du, Jiao; Xia, Fei; Lu, Jingli; Xiang, Ming

    2015-06-15

    Toll-like receptor 4 (TLR4) activation has been proposed to be important for islet cell inflammation and eventually ? cell loss in the course of type 1 diabetes (T1D) development. However, according to the "hygiene hypothesis", bacterial endotoxin lipopolysaccharide (LPS), an agonist on TLR4, inhibits T1D progression. Here we investigated possible mechanisms for the protective effect of LPS on T1D development in non-obese diabetic (NOD) mice. We found that LPS administration to NOD mice during the prediabetic state neither prevented nor reversed insulitis, but delayed the onset and decreased the incidence of diabetes, and that a multiple-injection protocol is more effective than a single LPS intervention. Further, LPS administration suppressed spleen T lymphocyte proliferation, increased the generation of CD4(+)CD25(+)Foxp3(+) regulatory T cells (Tregs), reduced the synthesis of strong Th1 proinflammatory cytokines, and downregulated TLR4 and its downstream MyD88-dependent signaling pathway. Most importantly, multiple injections of LPS induced a potential tolerogenic dendritic cell (DC) subset with low TLR4 expression without influencing the DC phenotype. Explanting DCs from repeated LPS-treated NOD mice into NOD/SCID diabetic mice conferred sustained protective effects against the progression of diabetes in the recipients. Overall, these results suggest that multiple mechanisms are involved in the protective effects of LPS against the development of diabetes in NOD diabetic mice. These include Treg induction, down-regulation of TLR4 and its downstream MyD88-dependent signaling pathway, and the emergence of a potential tolerogenic DC subset. PMID:25896969

  3. Benzo[a]pyrene modulation of acute immunologic responses in Red Sea bream pretreated with lipopolysaccharide.

    Science.gov (United States)

    Bo, Jun; Gopalakrishnan, Singaram; Fan, Dan-Qing; Thilagam, Harikrishnan; Qu, Hai-Dong; Zhang, Nai; Chen, Fang-Yi; Wang, Ke-Jian

    2014-05-01

    The effects of polycyclic aromatic hydrocarbons (PAHs) have been reported to modulate the immune response in aquatic animals, but the collected information of their effects on fish immunity is so far ambiguous. This study demonstrated that Benzo[a]pyrene (BaP) exposure altered the expression pattern of an antimicrobial peptide hepcidin (PM-hepc) gene and the activities of some immune-associated parameters in the lipopolysaccharide (LPS)-challenged red sea bream (Pagrus major). It was observed that LPS could increase respiratory burst, lysozyme and antibacterial activity in P. major. However when the P. major was exposed to different concentrations of BaP (1, 4, or 8 ?g L(-1) ) for 14 days and then challenged with LPS there was no significant change in the lysozyme and antibacterial activity. It was further observed that LPS could induce the PM-hepc mRNA expression at 3, 6, and 12-h post-LPS challenge. However, when P. major was exposed first to BaP for 14 days and then challenged with LPS, the expression of PM-hepc mRNA was delayed in the liver until 24 h and not significantly induced until 48 and 96 h. The mRNA expression pattern was completely different from that only with LPS challenge, showing that BaP exposure changed the PM-hepc mRNA expression pattern of fish with LPS challenge. This study demonstrated that BaP exposure can weaken or inhibit the induction of lysozyme and antibacterial activity in the LPS-challenged P. major; conversely BaP exposure could enhance the mRNA expression of PM-hepc gene, indicating that the effect of BaP has different modulatory mechanism on hepcidin genes and immune-associated parameters. PMID:22610821

  4. Neuroinflammatory response to lipopolysaccharide is exacerbated in mice genetically deficient in cyclooxygenase-2

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    Bosetti Francesca

    2008-05-01

    Full Text Available Abstract Background Cyclooxygenases (COX -1 and -2 are key mediators of the inflammatory response in the central nervous system. Since COX-2 is inducible by inflammatory stimuli, it has been traditionally considered as the most appropriate target for anti-inflammatory drugs. However, the specific roles of COX-1 and COX-2 in modulating a neuroinflammatory response are unclear. Recently, we demonstrated that COX-1 deficient mice show decreased neuroinflammatory response and neuronal damage in response to lipopolysaccharide (LPS. Methods In this study, we investigated the role of COX-2 in the neuroinflammatory response to intracerebroventricular-injected LPS (5 ?g, a model of direct activation of innate immunity, using COX-2 deficient (COX-2-/- and wild type (COX-2+/+ mice, as well as COX-2+/+ mice pretreated for 6 weeks with celecoxib, a COX-2 selective inhibitor. Results Twenty-four hours after LPS injection, COX-2-/- mice showed increased neuronal damage, glial cell activation, mRNA and protein expression of markers of inflammation and oxidative stress, such as cytokines, chemokines, iNOS and NADPH oxidase. Brain protein levels of IL-1?, NADPH oxidase subunit p67phox, and phosphorylated-signal transducer and activator of transcription 3 (STAT3 were higher in COX-2-/- and in celecoxib-treated mice, compared to COX-2+/+ mice. The increased neuroinflammatory response in COX-2-/- mice was likely mediated by the upregulation of STAT3 and suppressor of cytokine signaling 3 (SOCS3. Conclusion These results show that inhibiting COX-2 activity can exacerbate the inflammatory response to LPS, possibly by increasing glial cells activation and upregulating the STAT3 and SOCS3 pathways in the brain.

  5. Gene expression results in lipopolysaccharide-stimulated monocytes depend significantly on the choice of reference genes

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    Øvstebø Reidun

    2010-05-01

    Full Text Available Abstract Background Gene expression in lipopolysaccharide (LPS-stimulated monocytes is mainly studied by quantitative real-time reverse transcription PCR (RT-qPCR using GAPDH (glyceraldehyde 3-phosphate dehydrogenase or ACTB (beta-actin as reference gene for normalization. Expression of traditional reference genes has been shown to vary substantially under certain conditions leading to invalid results. To investigate whether traditional reference genes are stably expressed in LPS-stimulated monocytes or if RT-qPCR results are dependent on the choice of reference genes, we have assessed and evaluated gene expression stability of twelve candidate reference genes in this model system. Results Twelve candidate reference genes were quantified by RT-qPCR in LPS-stimulated, human monocytes and evaluated using the programs geNorm, Normfinder and BestKeeper. geNorm ranked PPIB (cyclophilin B, B2M (beta-2-microglobulin and PPIA (cyclophilin A as the best combination for gene expression normalization in LPS-stimulated monocytes. Normfinder suggested TBP (TATA-box binding protein and B2M as the best combination. Compared to these combinations, normalization using GAPDH alone resulted in significantly higher changes of TNF-? (tumor necrosis factor-alpha and IL10 (interleukin 10 expression. Moreover, a significant difference in TNF-? expression between monocytes stimulated with equimolar concentrations of LPS from N. meningitides and E. coli, respectively, was identified when using the suggested combinations of reference genes for normalization, but stayed unrecognized when employing a single reference gene, ACTB or GAPDH. Conclusions Gene expression levels in LPS-stimulated monocytes based on RT-qPCR results differ significantly when normalized to a single gene or a combination of stably expressed reference genes. Proper evaluation of reference gene stabiliy is therefore mandatory before reporting RT-qPCR results in LPS-stimulated monocytes.

  6. Designed beta-boomerang antiendotoxic and antimicrobial peptides: structures and activities in lipopolysaccharide.

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    Bhunia, Anirban; Mohanram, Harini; Domadia, Prerna N; Torres, Jaume; Bhattacharjya, Surajit

    2009-08-14

    Lipopolysaccharide (LPS), an integral part of the outer membrane of Gram-negative bacteria, is involved in a variety of biological processes including inflammation, septic shock, and resistance to host-defense molecules. LPS also provides an environment for folding of outer membrane proteins. In this work, we describe the structure-activity correlation of a series of 12-residue peptides in LPS. NMR structures of the peptides derived in complex with LPS reveal boomerang-like beta-strand conformations that are stabilized by intimate packing between the two aromatic residues located at the 4 and 9 positions. This structural feature renders these peptides with a high ability to neutralize endotoxicity, >80% at 10 nM concentration, of LPS. Replacements of these aromatic residues either with Ala or with Leu destabilizes the boomerang structure with the concomitant loss of antiendotoxic and antimicrobial activities. Furthermore, the aromatic packing stabilizing the beta-boomerang structure in LPS is found to be maintained even in a truncated octapeptide, defining a structured LPS binding motif. The mode of action of the active designed peptides correlates well with their ability to perturb LPS micelle structures. Fourier transform infrared spectroscopy studies of the peptides delineate beta-type conformations and immobilization of phosphate head groups of LPS. Trp fluorescence studies demonstrated selective interactions with LPS and the depth of insertion into the LPS bilayer. Our results demonstrate the requirement of LPS-specific structures of peptides for endotoxin neutralizations. In addition, we propose that structures of these peptides may be employed to design proteins for the outer membrane. PMID:19520860

  7. Lipopolysaccharide, Tumor Necrosis Factor Alpha, or Interleukin-1? Triggers Reactivation of Latent Cytomegalovirus in Immunocompetent Mice

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    Cook, Charles H.; Trgovcich, Joanne; Zimmerman, Peter D.; Zhang, Yingxue; Sedmak, Daniel D.

    2006-01-01

    We have previously shown that cytomegalovirus (CMV) can reactivate in lungs of nonimmunosuppressed patients during critical illness. Our recent work has shown that polymicrobial bacterial sepsis can trigger reactivation of latent murine CMV (MCMV). We hypothesize that MCMV reactivation following bacterial sepsis may be caused by inflammatory mediators. To test this hypothesis, BALB/c mice latently infected with Smith strain MCMV received sublethal intraperitoneal doses of lipopolysaccharide (LPS), tumor necrosis factor alpha (TNF-?), interleukin-1? (IL-1?), or saline. Lung tissue homogenates were evaluated for viral reactivation 3 weeks after mediator injection. Because LPS is known to signal via Toll-like receptor 4 (TLR-4) in mice, further studies blocking this signaling mechanism were performed using monoclonal MTS510. Finally, mice were tested with intravenous TNF-? to determine whether this would cause reactivation. All mice receiving sublethal intraperitoneal doses of LPS, TNF-?, or IL-1? had pulmonary reactivation of latent MCMV 3 weeks following injection, and LPS caused MCMV reactivation with kinetics similar to those for sepsis. When TLR-4 signaling was blocked, exogenous LPS did not reactivate latent MCMV. Intravenous TNF-? administration at near-lethal doses did not reactivate MCMV. Exogenous intraperitoneal LPS, TNF-?, and IL-1? are all capable of reactivating CMV from latency in lungs of previously healthy mice. LPS reactivation of MCMV appears dependent on TLR-4 signaling. Interestingly, intravenous TNF-? did not trigger reactivation, suggesting possible mechanistic differences that are discussed. We conclude that inflammatory disease states besides sepsis may be capable of reactivating CMV from latency. PMID:16940526

  8. Development of immunochromatography-based methods for detection of leptospiral lipopolysaccharide antigen in urine.

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    Widiyanti, Dian; Koizumi, Nobuo; Fukui, Takashi; Muslich, Lisa T; Segawa, Takaya; Villanueva, Sharon Y A M; Saito, Mitsumasa; Masuzawa, Toshiyuki; Gloriani, Nina G; Yoshida, Shin-ichi

    2013-05-01

    Leptospirosis is an infectious disease caused by the spirochete bacteria Leptospira spp. and is commonly found throughout the world. Diagnosis of leptospirosis performed by culture and microscopic agglutination tests is laborious and time-consuming. Therefore, we aimed to develop a novel immunochromatography (ICG)-based method for detecting Leptospira antigen in the urine of patients and animals. We used the 1H6 monoclonal antibody (MAb), which is specific to the lipopolysaccharide (LPS) that is common among Leptospira spp. The MAb was coupled to 40-nm-diameter colloidal gold, and the amounts of labeled antibody and immobilized antibody were 23 ?g and 2 ?g per test, respectively. Several strains of Leptospira and other bacterial species were used to evaluate the sensitivities and specificities of the assays we developed. The detection limit of the assays was 10(6) cells/ml when disrupted whole bacterial cells were used. The assays were Leptospira specific since they did not cross-react with non-Leptospira bacteria used in the study. Application of diagnostic assays was done on the urine samples of 46 Leptospira-infected hamsters, 44 patients with suspected leptospirosis, and 14 healthy individuals. Pretreatment of the urine samples by boiling and centrifugation (for ultrafiltration and concentration) eliminated nonspecific reactions that occurred in the assay. The sensitivity and specificity of the ICG-based lateral flow assay (LFA) were 89% and 87%, respectively, which were higher than those of the dipstick assay, which were 80% and 74%, respectively. In summary, this ICG-based LFA can be used as an alternative diagnostic assay for leptospirosis. Further development is still necessary to improve the assay. PMID:23467776

  9. Effect of bacterial lipopolysaccharide on gastric emptying of liquids in rats

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    E.F. Collares

    1997-02-01

    Full Text Available The objectives of the present investigation were 1 to study the effect of bacterial lipopolysaccharide (LPS on rat gastric emptying (GE and 2 to investigate a possible involvement of the vagus nerve in the gastric action of LPS. Endotoxin from E. coli (strain 055:B5 was administered sc, ip or iv to male Wistar rats (220-280 g body weight at a maximum dose of 50 µg/kg animal weight. Control animals received an equivalent volume of sterile saline solution. At a given time period after LPS administration, GE was evaluated by measuring gastric retention 10 min after the orogastric infusion of a test meal (2 ml/100 g animal weight, which consisted of 0.9% NaCl plus the marker phenol red (6 mg/dl. One group of animals was subjected to bilateral subdiaphragmatic vagotomy or sham operation 15 days before the test. A significant delay in GE of the test meal was observed 5 h after iv administration of the endotoxin at the dose of 50 µg/kg animal weight. The LPS-induced delay of GE was detected as early as 30 min and up to 8 h after endotoxin administration. The use of different doses of LPS ranging from 5 to 50 µg/kg animal weight showed that the alteration of GE was dose dependent. In addition, vagotomized animals receiving LPS displayed a GE that was not significantly different from that of the sham control group. However, a participation of the vagus nerve in LPS-induced delay in GE could not be clearly demonstrated by these experiments since vagotomy itself induced changes in this gastric parameter. The present study provides a suitable model for identifying the mechanisms underlying the effects of LPS on gastric emptying

  10. The effect of bacterial lipopolysaccharide on gastric emptying in rats suffering from moderate renal insufficiency

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    S.Z.P. Rigatto

    1998-04-01

    Full Text Available The objective of the present study was to evaluate the response of rats suffering from moderate renal insufficiency to bacterial lipopolysaccharide (LPS, or endotoxin. The study involved 48 eight-week-old male SPF Wistar rats (175-220 g divided into two groups of 24 animals each. One group underwent 5/6 nephrectomy while the other was sham-operated. Two weeks after surgery, the animals were further divided into two subgroups of 12 animals each and were fasted for 20 h but with access to water ad libitum. One nephrectomized and one sham-treated subgroup received E. coli LPS (25 µg/kg, iv while the other received a sterile, pyrogen-free saline solution. Gastric retention (GR was determined 10 min after the orogastric infusion of a standard saline test meal labeled with phenol red (6 mg/dl. The gastric emptying of the saline test meal was studied after 2 h. Renal function was evaluated by measuring the plasma levels of urea and creatinine. The levels of urea and creatinine in 5/6 nephrectomized animals were two-fold higher than those observed in the sham-operated rats. Although renal insufficiency did not change gastric emptying (median %GR = 26.6 for the nephrectomized subgroup and 29.3 for the sham subgroup, LPS significantly retarded the gastric emptying of the sham and nephretomized groups (median %GR = 42.0 and 61.0, respectively, and was significantly greater (P<0.01 in the nephrectomized rats. We conclude that gastric emptying in animals suffering from moderate renal insufficiency is more sensitive to the action of LPS than in sham animals

  11. Effect of bacterial lipopolysaccharide on gastric emptying of liquids in rats

    Scientific Electronic Library Online (English)

    E.F., Collares.

    1997-02-01

    Full Text Available The objectives of the present investigation were 1) to study the effect of bacterial lipopolysaccharide (LPS) on rat gastric emptying (GE) and 2) to investigate a possible involvement of the vagus nerve in the gastric action of LPS. Endotoxin from E. coli (strain 055:B5) was administered sc, ip or i [...] v to male Wistar rats (220-280 g body weight) at a maximum dose of 50 µg/kg animal weight. Control animals received an equivalent volume of sterile saline solution. At a given time period after LPS administration, GE was evaluated by measuring gastric retention 10 min after the orogastric infusion of a test meal (2 ml/100 g animal weight), which consisted of 0.9% NaCl plus the marker phenol red (6 mg/dl). One group of animals was subjected to bilateral subdiaphragmatic vagotomy or sham operation 15 days before the test. A significant delay in GE of the test meal was observed 5 h after iv administration of the endotoxin at the dose of 50 µg/kg animal weight. The LPS-induced delay of GE was detected as early as 30 min and up to 8 h after endotoxin administration. The use of different doses of LPS ranging from 5 to 50 µg/kg animal weight showed that the alteration of GE was dose dependent. In addition, vagotomized animals receiving LPS displayed a GE that was not significantly different from that of the sham control group. However, a participation of the vagus nerve in LPS-induced delay in GE could not be clearly demonstrated by these experiments since vagotomy itself induced changes in this gastric parameter. The present study provides a suitable model for identifying the mechanisms underlying the effects of LPS on gastric emptying

  12. Ginsenoside rg2 inhibits lipopolysaccharide-induced adhesion molecule expression in human umbilical vein endothelial cell.

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    Cho, Young-Suk; Kim, Chan Hyung; Ha, Tae-Sun; Lee, Sang Jin; Ahn, Hee Yul

    2013-04-01

    Vascular cell adhesion molecule 1 (VCAM-1), intercellular adhesion molecule 1 (ICAM-1), P- and E-selectin play a pivotal role for initiation of atherosclerosis. Ginsenoside, a class of steroid glycosides, is abundant in Panax ginseng root, which has been used for prevention of illness in Korea. In this study, we investigated the mechanism(s) by which ginsenoside Rg2 may inhibit VCAM-1 and ICAM-1 expressions stimulated with lipopolysaccharide (LPS) in human umbilical vein endothelial cell (HUVEC). LPS increased VCAM-1 and ICAM-1 expression. Ginsenoside Rg2 prevented LPS-mediated increase of VCAM-1 and ICAM-1 expression. On the other hand, JSH, a nuclear factor kappa B (NF-?B) inhibitor, reduced both VCAM-1 and ICAM-1 expression stimulated with LPS. SB202190, inhibitor of p38 mitogen-activated protein kinase (p38 MAPK), and wortmannin, phosphatidylinositol 3-kinase (PI3-kinase) inhibitor, reduced LPS-mediated VCAM-1 but not ICAM-1 expression. PD98059, inhibitor of mitogen-activated protein kinase kinase/extracellular signal-regulated kinase (MEK/ERK) did not affect VCAM-1 and ICAM-1 expression stimulated with LPS. SP600125, inhibitor of c-Jun N-terminal kinase (JNK), reduced LPS-mediated ICAM-1 but not VCAM-1 expression. LPS reduced IkappaB? (I?B?) expression, in a time-dependent manner within 1 hr. Ginsenoside Rg2 prevented the decrease of I?B? expression stimulated with LPS. Moreover, ginsenoside Rg2 reduced LPS-mediated THP-1 monocyte adhesion to HUVEC, in a concentration-dependent manner. These data provide a novel mechanism where the ginsenoside Rg2 may provide direct vascular benefits with inhibition of leukocyte adhesion into vascular wall thereby providing protection against vascular inflammatory disease. PMID:23626475

  13. The roles of prostaglandin E2 and D2 in lipopolysaccharide-mediated changes in sleep.

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    Oishi, Yo; Yoshida, Kyoko; Scammell, Thomas E; Urade, Yoshihiro; Lazarus, Michael; Saper, Clifford B

    2015-07-01

    When living organisms become sick as a result of a bacterial infection, a suite of brain-mediated responses occur, including fever, anorexia and sleepiness. Systemic administration of lipopolysaccharide (LPS), a common constituent of bacterial cell walls, increases body temperature and non-rapid eye movement (NREM) sleep in animals and induces the production of pro-inflammatory prostaglandins (PGs). PGE2 is the principal mediator of fever, and both PGE2 and PGD2 regulate sleep-wake behavior. The extent to which PGE2 and PGD2 are involved in the effect of LPS on NREM sleep remains to be clarified. Therefore, we examined LPS-induced changes in body temperature and NREM sleep in mice with nervous system-specific knockouts (KO) for the PGE2 receptors type EP3 or EP4, in mice with total body KO of microsomal PGE synthase-1 or the PGD2 receptor type DP, and in mice treated with the cyclooxygenase (COX) inhibitor meloxicam. We observed that LPS-induced NREM sleep was slightly attenuated in mice lacking EP4 receptors in the nervous system, but was not affected in any of the other KO mice or in mice pretreated with the COX inhibitor. These results suggest that the effect of LPS on NREM sleep is partially dependent on PGs and is likely mediated mainly by other pro-inflammatory substances. In addition, our data show that the main effect of LPS on body temperature is hypothermia in the absence of nervous system EP3 receptors or in the presence of a COX inhibitor. PMID:25532785

  14. Computer simulation of uranyl uptake by the rough lipopolysaccharide membrane of Pseudomonas aeruginosa.

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    Lins, Roberto D; Vorpagel, Erich R; Guglielmi, Matteo; Straatsma, T P

    2008-01-01

    Heavy metal environmental contaminants cannot be destroyed but require containment, preferably in concentrated form, in a solid or immobile form for recycling or final disposal. Microorganisms are able to take up and deposit high levels of contaminant metals, including radioactive metals such as uranium and plutonium, into their cell wall. Consequently, these microbial systems are of great interest as the basis for potential environmental bioremediation technologies. The outer membranes of Gram-negative microbes are highly nonsymmetric and exhibit a significant electrostatic potential gradient across the membrane. This gradient has a significant effect on the uptake and transport of charged and dipolar compounds. However, the effectiveness of microbial systems for environmental remediation will depend strongly on specific properties that determine the uptake of targeted contaminants by a particular cell wall. To aid in the design of microbial remediation technologies, knowledge of the factors that determine the affinity of a particular bacterial outer membrane for the most common ionic species found in contaminated soils and groundwater is of great importance. Using our previously developed model for the lipopolysaccharide (LPS) membrane of Pseudomonas aeruginosa, this work presents the potentials of mean force as the estimate of the free energy profile for uptake of sodium, calcium, chloride, uranyl ions, and a water molecule by the bacterial LPS membrane. A compatible classical parameter set for uranyl has been developed and validated. Results show that the uptake of uranyl is energetically a favorable process relative to the other ions studied. At neutral pH, this nuclide is shown to be retained on the surface of the LPS membrane through chelation with the carboxyl and hydroxyl groups located in the outer core. PMID:18067253

  15. In silico biosynthesis of virenose, a methylated deoxy-sugar unique to Coxiella burnetii lipopolysaccharide

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    Flores-Ramirez Gabriela

    2012-11-01

    Full Text Available Abstract Background Coxiella burnetii is Gram-negative bacterium responsible for the zoonosis Q-fever. While it has an obligate intracellular growth habit, it is able to persist for extended periods outside of a host cell and can resist environmental conditions that would be lethal to most prokaryotes. It is these extracellular bacteria that are the infectious stage encountered by eukaryotic hosts. The intracellular form has evolved to gro