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Sample records for porphyromonas gingivalis lipopolysaccharide

  1. Hemin-induced modifications of the antigenicity and hemin-binding capacity of Porphyromonas gingivalis lipopolysaccharide.

    OpenAIRE

    Cutler, C W; Eke, P I; Genco, C. A.; Van Dyke, T E; Arnold, R R

    1996-01-01

    Previous studies have shown that the physical, biochemical, and antigenic properties of the bacterial outer membrane are profoundly influenced by the growth environment. In the present study, the effects of growth in hemin-replete (H+) and hemin-depleted (H-) media on the lipopolysaccharide (LPS) of the oral pathogen Porphyromonas gingivalis were investigated. Our studies show that LPS from P. gingivalis cultured in H+ media (H+LPS) expressed additional low-molecular-mass antigens, as determi...

  2. Xylitol Inhibits Inflammatory Cytokine Expression Induced by Lipopolysaccharide from Porphyromonas gingivalis

    OpenAIRE

    Han, Su-Ji; Jeong, So-Yeon; Nam, Yun-Ju; Yang, Kyu-Ho; Lim, Hoi-Soon; Chung, Jin

    2005-01-01

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

  3. Effect of Neutrophil Apoptosis on Monocytic Cytokine Response to Porphyromonas gingivalis Lipopolysaccharide

    Science.gov (United States)

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

    2005-01-01

    Background Neutrophil apoptosis may play a critical role in the resolution of inflammation by stimulating anti-inflammatory cytokine generation from monocytes. In this study, we investigated the effect of apoptotic neutrophils on interleukin (IL)-10 and IL-1? production from monocytes in response to Porphyromonas gingivalis lipopolysaccharide. Methods Peripheral blood neutrophils from healthy individuals were isolated by sodium diatrizoate density gradient centrifugation. In order to induce apoptosis, neutrophils were cultured for 24 hours in modified Dulbecco’s medium supplemented with 10% autologous serum. Cell apoptosis was quantified by Annexin V positivity and loss of CD16 expression on the cell surface. Peripheral blood mononuclear cells were isolated from the same subjects; monocytes were purified by magnetic cell sorting and cultured with or without apoptotic or fresh neutrophils. Lipopolysaccharide from Porphyromonas gingivalis was used for cell stimulation. IL-1? and IL-10 levels in supernatants were determined by enzyme-linked immunosorbent assay (ELISA). Results IL-10 generation was significantly increased in monocytes cultured with apoptotic neutrophils compared to monocytes alone or cocultured with fresh neutrophils (P <0.05). IL-1? was suppressed both in resting and lipopolysaccharide-stimulated monocytes in the presence of apoptotic neutrophils compared to monocytes alone or monocytes cultured with fresh neutrophils at all time points (P <0.05). Conclusion Neutrophil apoptosis provides a signal to monocytes, changing the phenotype of the monocyte resulting in the production of anti-inflammatory cytokines and suppression of proinflammatory cytokines in response to lipopolysaccharide. PMID:15948692

  4. 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.

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

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    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.

  6. 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.

  7. Bortezomib Inhibits Osteoclastogenesis and Porphyromonas gingivalis Lipopolysaccharide-induced Alveolar Bone Resorption.

    Science.gov (United States)

    Kim, Y-G; Kang, J H; Kim, H J; Kim, H J; Kim, H-H; Kim, J-Y; Lee, Y

    2015-09-01

    Healthy bone is maintained by the coordinated activities of osteoblast-mediated bone formation and osteoclast-dependent bone resorption. Pathologic conditions such as hormonal imbalance and inflammation cause increased osteoclastogenesis resulting in osteoporosis, rheumatoid arthritis, and periodontitis. Bortezomib is novel antimyeloma agent that has a direct beneficial effect on bone formation. However, the role of bortezomib in osteoclastogenesis and underlying mechanisms remains to be fully comprehended. In the present study, we show that bortezomib directly inhibited the receptor activator of nuclear factor ?B ligand (RANKL)- and lipopolysaccharide-dependent osteoclast differentiation. Interestingly, the bortezomib-mediated inhibition of osteoclastogenesis was transient, since the removal of bortezomib from culture completely restored osteoclast differentiation. Bortezomib impeded the induction and nuclear localization of nuclear factor of activated T cells, cytoplasmic 1 and reduced both macrophage colony-stimulating factor- and RANKL-induced extracellular-signal-regulated kinase (ERK) phosphorylation. In a mouse model of periodontitis, bortezomib prevented alveolar bone erosion induced by Porphyromonas gingivalis lipopolysaccharide. These data not only suggest a previously unappreciated mechanism by which bortezomib regulates bone resorption but also propose novel applications of bortezomib beyond its use as an antimyeloma agent. PMID:26130255

  8. Porphyromonas gingivalis: a clonal pathogen?

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    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.

  9. Antibacterial Action of Polyphosphate on Porphyromonas gingivalis?

    OpenAIRE

    Moon, Ji-Hoi; Park, Jae-Hong; Lee, Jin-yong

    2010-01-01

    Polyphosphate [poly(P)] has antibacterial activity against various Gram-positive bacteria. In contrast, Gram-negative bacteria are generally resistant to poly(P). Here, we describe the antibacterial characterization of poly(P) against a Gram-negative periodontopathogen, Porphyromonas gingivalis. The MICs of pyrophosphate (Na4P2O7) and all poly(P) (Nan + 2PnO3n + 1; n = 3 to 75) tested for the bacterium by the agar dilution method were 0.24% and 0.06%, respectively. Orthophosphate (Na2HPO4) fa...

  10. Presence of Porphyromonas gingivalis in gingival squamous cell carcinoma

    OpenAIRE

    Katz, Joseph; Onate, Mairelys D; Pauley, Kaleb M; 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. Identification of a second endogenous Porphyromonas gingivalis insertion element.

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    Wang, C.Y.; Bond, V C; Genco, C A

    1997-01-01

    In this study a second endogenous Porphyromonas gingivalis insertion element (IS element) that is capable of transposition within P. gingivalis was identified. Nucleotide sequence analysis of the Tn4351 insertion site in a P. gingivalis Tn4351-generated transconjugant showed that a complete copy of the previously unidentified IS element, designated PGIS2, had inserted into IS4351R in Tn4351. PGIS2 is 1,207 bp in length with 19-bp imperfect terminal inverted repeats, and insertion resulted in ...

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

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

    2013-01-01

    Porphyromonas gingivalis is considered as a major etiological agent in periodontal diseases and implied to result in gingival inflammation under orthodontic appliance. rag locus is a pathogenicity island found in Porphyromonas gingivalis. Four rag locus variants are different in pathogenicity of Porphyromonas gingivalis. Moreover, there are different racial and geographic differences in distribution of rag locus genotypes. In this study, we assessed the prevalence of Porphyromonas gingivalis ...

  13. 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 enfermedades loca...

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

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    Le Xuan

    2009-01-01

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

  15. Invasion of Porphyromonas gingivalis strains into vascular cells and tissue

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    Ingar Olsen

    2015-08-01

    Full Text Available Porphyromonas gingivalis is considered a major pathogen in adult periodontitis and is also associated with multiple systemic diseases, for example, cardiovascular diseases. One of its most important virulence factors is invasion of host cells. The invasion process includes attachment, entry/internalization, trafficking, persistence, and exit. The present review discusses these processes related to P. gingivalis in cardiovascular cells and tissue. Although most P. gingivalis strains invade, the invasion capacity of strains and the mechanisms of invasion including intracellular trafficking among them differ. This is consistent with the fact that there are significant differences in the pathogenicity of P. gingivalis strains. P. gingivalis invasion mechanisms are also dependent on types of host cells. Although much is known about the invasion process of P. gingivalis, we still have little knowledge of its exit mechanisms. Nevertheless, it is intriguing that P. gingivalis can remain viable in human cardiovascular cells and atherosclerotic plaque and later exit and re-enter previously uninfected host cells.

  16. LuxS signaling in Porphyromonas gingivalis-host interactions.

    Science.gov (United States)

    Scheres, Nina; Lamont, Richard J; Crielaard, Wim; Krom, Bastiaan P

    2015-10-01

    Dental plaque is a multispecies biofilm in the oral cavity that significantly influences oral health. The presence of the oral anaerobic pathogen Porphyromonas gingivalis is an important determinant in the development of periodontitis. Direct and indirect interactions between P. gingivalis and the host play a major role in disease development. Transcriptome analysis recently revealed that P. gingivalis gene-expression is regulated by LuxS in both an AI-2-dependent and an AI-2 independent manner. However, little is known about the role of LuxS-signaling in P. gingivalis-host interactions. Here, we investigated the effect of a luxS mutation on the ability of P. gingivalis to induce an inflammatory response in human oral cells in vitro. Primary periodontal ligament (PDL) fibroblasts were challenged with P. gingivalis ?luxS or the wild-type parental strain and gene-expression of pro-inflammatory mediators IL-1?, IL-6 and MCP-1 was determined by real-time PCR. The ability of P. gingivalis ?luxS to induce an inflammatory response was severely impaired in PDL-fibroblasts. This phenotype could be restored by providing of LuxS in trans, but not by addition of the AI-2 precursor DPD. A similar phenomenon was observed in a previous transcriptome study showing that expression of PGN_0482 was reduced in the luxS mutant independently of AI-2. We therefore also analyzed the effect of a mutation in PGN_0482, which encodes an immuno-reactive, putative outer-membrane protein. Similar to P. gingivalis ?luxS, the P. gingivalis ?0482 mutant had an impaired ability to induce an inflammatory response in PDL fibroblasts. LuxS thus appears to influence the pro-inflammatory responses of host cells to P. gingivalis, likely through regulation of PGN_0482. PMID:25434960

  17. Attenuation of Porphyromonas gingivalis oral infection by ?-amylase and pentamidine.

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    Li, Ying; Miao, Yu-Song; Fu, Yun; Li, Xi-Ting; Yu, Shao-Jie

    2015-08-01

    The Porphyromonas gingivalis bacterium is one of the most influential pathogens in oral infections. In the current study, the antimicrobial activity of ?-amylase and pentamidine against Porphyromonas gingivalis was evaluated. Their in vitro inhibitory activity was investigated with the agar overlay technique, and the minimal inhibitory and bactericidal concentrations were determined. Using the bactericidal concentration, the antimicrobial actions of the inhibitors were investigated. In the present study, multiple techniques were utilized, including scanning electron microscopy (SEM), general structural analysis and differential gene expression analysis. The results obtained from SEM and bactericidal analysis indicated a notable observation; the pentamidine and ?-amylase treatment destroyed the structure of the bacterial cell membranes, which led to cell death. These results were used to further explore these inhibitors and the mechanisms by which they act. Downregulated expression levels were observed for a number of genes coding for hemagglutinins and gingipains, and various genes involved in hemin uptake, chromosome replication and energy production. However, the expression levels of genes associated with iron storage and oxidative stress were upregulated by ?-amylase and pentamidine. A greater effect was noted in response to pentamidine treatment. The results of the present study demonstrate promising therapeutic potential for ?-amylases and pentamidine. These molecules have the potential to be used to develop novel drugs and broaden the availability of pharmacological tools for the attenuation of oral infections caused by Porphyromonas gingivalis. PMID:25846026

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

    OpenAIRE

    Le Xuan; Laflamme Claude; Rouabhia Mahmoud

    2009-01-01

    Background: Porphyromonas gingivalis, an important periodontal pathogen, is closely associated with inflammatory alveolar bone resorption. This bacterium exerts its pathogenic effect indirectly through multiple virulence factors, such as lipopolysaccharides, fimbriae, and proteases. Another possible pathogenic path may be through a direct interaction with the host?s soft and hard tissues (e.g., alveolar bone), which could lead to periodontitis. Aims and Objectives: The aim of the ...

  19. Phagocytosis of virulent Porphyromonas gingivalis by human polymorphonuclear leukocytes requires specific immunoglobulin G.

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    Cutler, C W; Kalmar, J R; Arnold, R R

    1991-01-01

    No studies to date clearly define the interactions between Porphyromonas gingivalis and human peripheral blood polymorphonuclear leukocytes (PMN), nor has a protective role for antibody to P. gingivalis been defined. Using a fluorochrome phagocytosis microassay, we investigated PMN phagocytosis and killing of P. gingivalis as a function of P. gingivalis-specific antibody. Sera from a nonimmune rabbit and a healthy human subject were not opsonic for virulent P. gingivalis A7436, W83, and HG405...

  20. Identification of a Porphyromonas gingivalis Receptor for the Streptococcus gordonii SspB Protein

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    Chung, Whasun O; Demuth, Donald R.; Lamont, Richard J.

    2000-01-01

    Colonization of the plaque biofilm by the oral pathogen Porphyromonas gingivalis is favored by the presence of antecedent organisms such as Streptococcus gordonii. Coadhesion between P. gingivalis and S. gordonii can be mediated by the SspB protein of S. gordonii; however, the P. gingivalis cognate receptor for this protein has not been identified. In this study, we identified a surface protein of P. gingivalis that interacts with the SspB protein. Coprecipitation between P. gingivalis outer ...

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

    Science.gov (United States)

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

    1993-03-01

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

  2. Binding and accumulation of hemin in Porphyromonas gingivalis are induced by hemin.

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    Genco, C. A.; Odusanya, B M; Brown, G.

    1994-01-01

    Although hemin is an essential nutrient for the black-pigmented oral bacterium Porphyromonas gingivalis, the mechanisms involved in hemin binding and uptake are poorly defined. In this study, we have examined the binding of hemin and Congo red (CR) to P. gingivalis whole cells and have defined the conditions for maximal binding. Additionally, the accumulation of hemin by P. gingivalis under growing conditions has been characterized. P. gingivalis A7436 was grown under hemin- or iron-deplete c...

  3. The Porphyromonas gingivalis Ferric Uptake Regulator Orthologue Binds Hemin and Regulates Hemin-Responsive Biofilm Development

    OpenAIRE

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

    2014-01-01

    Porphyromonas gingivalis is a Gram-negative pathogen associated with the biofilm-mediated disease chronic periodontitis. P. gingivalis biofilm formation is dependent on environmental heme for which P. gingivalis has an obligate requirement as it is unable to synthesize protoporphyrin IX de novo, hence P. gingivalis transports iron and heme liberated from the human host. Homeostasis of a variety of transition metal ions is often mediated in Gram-negative bacteria at the transcriptional level b...

  4. 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.

  5. Complete Genome Sequence of the Bacterium Porphyromonas gingivalis TDC60, Which Causes Periodontal Disease?

    OpenAIRE

    Watanabe, Takayasu; Maruyama, Fumito; Nozawa, Takashi (JAEA); Aoki, Akinobu; Okano, Soichiro; Shibata, Yasuko; Oshima, Kenshiro; Kurokawa, Ken; Hattori, Masahira; NAKAGAWA, ICHIRO; Abiko, Yoshimitsu

    2011-01-01

    Porphyromonas gingivalis is a black-pigmented asaccharolytic anaerobe and a major causative agent of periodontitis. Here, we report the complete genome sequence of P. gingivalis strain TDC60, which was recently isolated from a severe periodontal lesion in a Japanese patient.

  6. Influence of immunization on Porphyromonas gingivalis colonization and invasion in the mouse chamber model.

    OpenAIRE

    Genco, C. A.; Kapczynski, D R; Cutler, C W; Arko, R J; Arnold, R R

    1992-01-01

    The effects of immunization with invasive or noninvasive Porphyromonas (Bacteroides) gingivalis strains on the pathogenesis of infection in a mouse chamber model were examined. BALB/c mice were immunized by a single injection of heat-killed P. gingivalis invasive strain A7436 or W83 or noninvasive strain 33277, HG405, or 381 directly into subcutaneous chambers. P. gingivalis-specific antibody was detected in chamber fluid 21 days postimmunization, and mice were subsequently challenged by inje...

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

    OpenAIRE

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

    1995-01-01

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

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

    DEFF Research Database (Denmark)

    Belstrøm, Daniel; Holmstrup, Palle; Damgaard, Christian; Borch, Tanja Skuldbøl; Skjødt, Mikkel-Ole; Bendtzen, Klaus; Nielsen, Claus Kim Hostein

    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 ...

  9. 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

  10. A novel mouse model to study the virulence of and host response to Porphyromonas (Bacteroides) gingivalis.

    OpenAIRE

    Genco, C. A.; Cutler, C W; Kapczynski, D; Maloney, K; Arnold, R R

    1991-01-01

    We describe here the development of a mouse subcutaneous chamber model that allows for the examination of host-parasite interactions as well as the determination of gross pathology with Porphyromonas (Bacteroides) gingivalis challenge. When inoculated into stainless-steel chambers implanted subcutaneously in female BALB/c mice, P. gingivalis W83, W50, and A7436 (10(8) to 10(10) CFU) caused cachexia, ruffling, general erythema and phlegmonous, ulcerated, necrotic lesions, and death. P. gingiva...

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

    OpenAIRE

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

    2012-01-01

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

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

    OpenAIRE

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

    1994-01-01

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

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

    OpenAIRE

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

    1999-01-01

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

  14. Porphyromonas gingivalis SerB mediated dephosphorylation of host cell cofilin modulates invasion efficiency

    OpenAIRE

    Moffatt, Catherine E.; Inaba, Hiroaki; Hirano, Takanori; Lamont, Richard J.

    2012-01-01

    Porphyromonas gingivalis a host-adapted opportunistic pathogen produces a serine phosphatase, SerB, known to affect virulence, invasion and persistence within the host cell. SerB induces actin filament rearrangement in epithelial cells, but the mechanistic basis of this is not fully understood. Here we investigated the effects of SerB on the actin depolymerizing host protein cofilin. P. gingivalis infection resulted in the dephosphorylation of cofilin in gingival epithelial cells. In contrast...

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

    OpenAIRE

    XIE, HUA; Zheng, Cunge

    2012-01-01

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

  16. 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...

  17. 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.

  18. 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

  19. 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’.

  20. Identification of a Porphyromonas gingivalis receptor for the Streptococcus gordonii SspB protein.

    Science.gov (United States)

    Chung, W O; Demuth, D R; Lamont, R J

    2000-12-01

    Colonization of the plaque biofilm by the oral pathogen Porphyromonas gingivalis is favored by the presence of antecedent organisms such as Streptococcus gordonii. Coadhesion between P. gingivalis and S. gordonii can be mediated by the SspB protein of S. gordonii; however, the P. gingivalis cognate receptor for this protein has not been identified. In this study, we identified a surface protein of P. gingivalis that interacts with the SspB protein. Coprecipitation between P. gingivalis outer membrane proteins and purified SspB protein demonstrated that a 100-kDa P. gingivalis protein bound to SspB. The 100-kDa protein also bound to an engineered strain of Enterococcus faecalis that expresses the SspB protein on the cell surface. Monospecific polyclonal antibodies to the 100-kDa protein inhibited the binding between P. gingivalis and S. gordonii in a dose-dependent manner up to 86%. Amino acid sequencing of the 100-kDa protein showed homology to a protein previously identified as the P. gingivalis minor fimbria. The minor fimbrial protein may exist as a complex with a hemagglutinin-like protein since the genes encoding these proteins are adjacent on the chromosome and are cotranscribed. Thus, the P. gingivalis receptor for S. gordonii SspB is a 100-kDa protein that structurally may be a minor fimbria-protein complex and functionally effectuates coadhesion. PMID:11083792

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

    Science.gov (United States)

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

    2013-02-01

    The purpose of this study was to determine whether epigallocatechin-3-gallate (EGCG) ameliorates Porphyromonas gingivalis-induced atherosclerosis. EGCG is a polyphenol extract from green tea with health benefits and P. gingivalis is shown here to accelerate atheroma formation in a murine model. Apolipoprotein E knockout mice were administered EGCG or vehicle in drinking water; they were then fed high-fat diets and injected with P. gingivalis three times a week for 3 weeks. Mice were then killed at 15 weeks. Atherosclerotic plaques in the proximal aorta were determined by Oil Red O staining. Atherosclerosis risk factors in serum, liver or aorta were analysed using cytokine antibody arrays, enzyme-linked immunosorbent assay and real-time PCR. Atherosclerotic lesion areas of the aortic sinus caused by P. gingivalis infection decreased in EGCG-treated groups, wherein EGCG reduced the production of C-reactive protein, monocyte chemoattractant protein-1, and oxidized low-density lipoprotein (LDL), and slightly lowered LDL/very LDL cholesterol in P. gingivalis-challenged mice serum. Furthermore, the increase in CCL2, MMP-9, ICAM-1, HSP60, CD44, LOX-1, NOX-4, p22phox and iNOS gene expression levels in the aorta of P. gingivalis-challenged mice were reduced in EGCG-treated mice. However, HO-1 mRNA levels were elevated by EGCG treatment, suggesting that EGCG, as a natural substance, inhibits P. gingivalis-induced atherosclerosis through anti-inflammatory and antioxidative effects. PMID:23620122

  2. Adhesion Molecule Deficiencies Increase Porphyromonas gingivalis-Induced Alveolar Bone Loss in Mice

    OpenAIRE

    Baker, Pamela J.; DuFour, Lisa; Dixon, Mark; Roopenian, Derry. C.

    2000-01-01

    Alveolar bone resorption can be induced in specific-pathogen-free mice by oral infection with Porphyromonas gingivalis (P. J. Baker, R. T. Evans, and D. C. Roopenian, Arch. Oral Biol. 39:1035–1040, 1994). Here we used a mouse strain, C57BL/6J, which is relatively resistant to P. gingivalis-induced bone loss to examine whether partial or complete deletion of various adhesion molecules would increase susceptibility. Complete deletion of P-selectin or nearly complete lack of expression of interc...

  3. Hemin uptake in Porphyromonas gingivalis: Omp26 is a hemin-binding surface protein.

    OpenAIRE

    Bramanti, T E; Holt, S.C.

    1993-01-01

    A 26-kDa outer membrane protein (Omp26) has been proposed to play a role in hemin acquisition by Porphyromonas gingivalis (T. E. Bramanti and S. C. Holt, J. Bacteriol. 174:5827-5839, 1992). We studied [55Fe]hemin uptake in P. gingivalis grown under conditions of hemin starvation (Omp26 expressed on the outer membrane surface) and hemin excess (Omp26 not expressed on surface). [55Fe]hemin uptake occurred rapidly in hemin-starved cells which incorporated up to 70% of total [55Fe]hemin within 3 ...

  4. 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...

  5. LPS from Porphyromonas gingivalis increases the sensitivity of contractile response mediated by endothelin-B (ET(B)) receptors in cultured endothelium-intact rat coronary arteries

    DEFF Research Database (Denmark)

    Ghorbani, Bahareh; Holmstrup, Palle; Edvinsson, Lars; Kristiansen, Kim A; Sheykhzade, Majid

    2010-01-01

    The purpose of our study was to examine if lipopolysaccharide (LPS) from Porphyromonas gingivalis (P.g.) modifies the vasomotor responses to Endothelin-1 (ET-1) and Sarafotoxin 6c (S6c) in rat coronary arteries. The arteries were studied directly or following organ culture for 24h in absence and presence of 2.5EU/ml LPS. The contractile responses of coronary arteries were investigated by using the selective ETB receptor agonist S6c (1 pM-0.3µM) and ET-1 (1 pM-0.3µM). The functional studies demon...

  6. Porphyromonas gingivalis Stimulates Bone Resorption by Enhancing RANKL (Receptor Activator of NF-?B Ligand) through Activation of Toll-like Receptor 2 in Osteoblasts.

    Science.gov (United States)

    Kassem, Ali; Henning, Petra; Lundberg, Pernilla; Souza, Pedro P C; Lindholm, Catharina; Lerner, Ulf H

    2015-08-14

    Periodontitis has been associated with rheumatoid arthritis. In experimental arthritis, concomitant periodontitis caused by oral infection with Porphyromonas gingivalis enhances articular bone loss. The aim of this study was to investigate how lipopolysaccharide (LPS) from P. gingivalis stimulates bone resorption. The effects by LPS P. gingivalis and four other TLR2 ligands on bone resorption, osteoclast formation, and gene expression in wild type and Tlr2-deficient mice were assessed in ex vivo cultures of mouse parietal bones and in an in vivo model in which TLR2 agonists were injected subcutaneously over the skull bones. LPS P. gingivalis stimulated mineral release and matrix degradation in the parietal bone organ cultures by increasing differentiation and formation of mature osteoclasts, a response dependent on increased RANKL (receptor activator of NF-?B ligand). LPS P. gingivalis stimulated RANKL in parietal osteoblasts dependent on the presence of TLR2 and through a MyD88 and NF-?B-mediated mechanism. Similarly, the TLR2 agonists HKLM, FSL1, Pam2, and Pam3 stimulated RANKL in osteoblasts and parietal bone resorption. LPS P. gingivalis and Pam2 robustly enhanced osteoclast formation in periosteal/endosteal cell cultures by increasing RANKL. LPS P. gingivalis and Pam2 also up-regulated RANKL and osteoclastic genes in vivo, resulting in an increased number of periosteal osteoclasts and immense bone loss in wild type mice but not in Tlr2-deficient mice. These data demonstrate that LPS P. gingivalis stimulates periosteal osteoclast formation and bone resorption by stimulating RANKL in osteoblasts via TLR2. This effect might be important for periodontal bone loss and for the enhanced bone loss seen in rheumatoid arthritis patients with concomitant periodontal disease. PMID:26085099

  7. Antibody-dependent alternate pathway of complement activation in opsonophagocytosis of Porphyromonas gingivalis.

    OpenAIRE

    Cutler, C W; Kalmar, J R; Arnold, R R

    1991-01-01

    It has been suggested that the ability of Porphyromonas gingivalis to proteolyse complement, as well as its production of a capsule, contributes to resistance to phagocytosis by polymorphonuclear leukocytes. In this report, the opsonic role of serum complement and its activation pathways were investigated, using individual sera heat treated or depleted of factors B, C2, and C1q and the divalent cations Mg2+ and Ca2+. A fluorochrome microassay was used to quantitate phagocytosis of P. gingival...

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

    OpenAIRE

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

    2012-01-01

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

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

    OpenAIRE

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

    1997-01-01

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

  10. Elafin is Specifically inactivated by RgpB from Porphyromonas gingivalis by Distinct Proteolytic Cleavage

    OpenAIRE

    Kantyka, Tomasz; Latendorf, Ties; Wiedow, Oliver; Bartels, Joachim; Gläser, Regine; Dubin, Grzegorz; Schröder, Jens-Michael; Potempa, Jan; Meyer-Hoffert, Ulf

    2009-01-01

    Porphyromonas gingivalis, the major causative bacterium of periodontitis, contributes significantly to elevated proteolytic activity at periodontal pockets due to the presence of both, bacteria and host, predominantly neutrophil-derived, serine proteases. Normally the activity of the latter enzymes is tightly regulated by endogenous proteins, including elafin, a potent neutrophil elastase and proteinase 3 inhibitor released from epithelial cells at site of inflammation. Here we have found tha...

  11. Communication: Antimicrobial Activity of SMAP28 with a Targeting Domain for Porphyromonas gingivalis

    OpenAIRE

    Bratt, Carol L.; Kohlgraf, Karl G.; Yohnke, Katie; Kummet, Colleen; Dawson, Deborah V; Brogden, Kim A

    2010-01-01

    Antibiotic therapy is often used with mechanical therapy to treat periodontal disease. However, complications associated with antibiotic use can occur. A ‘bacteria-specific’ targeted approach would eliminate some of these complications and kill specific periodontopathogens without harming the commensal bacteria. One such approach is to couple antimicrobial peptides to a ligand, pheromone, or antibody specific for the periodontopathogen, Porphyromonas gingivalis. To assess the feasibility of t...

  12. Discrete Protein Determinant Directs the Species-Specific Adherence of Porphyromonas gingivalis to Oral Streptococci

    OpenAIRE

    Demuth, Donald R.; Irvine, Douglas C.; Costerton., J.W.; Cook, Guy S.; Lamont, Richard J.

    2001-01-01

    For pathogens to survive in the human oral cavity, they must identify a suitable niche in the complex multispecies biofilm that exists on oral tissues. The periodontal pathogen Porphyromonas gingivalis adheres to Streptococcus gordonii by interacting with a specific region of the streptococcal SspB polypeptide, designated BAR. However, it does not adhere to Streptococcus mutans, which expresses SpaP, a highly conserved homolog of SspB. Comparison of the predicted secondary structure of BAR wi...

  13. Purification and characterization of three types of proteases from culture supernatants of Porphyromonas gingivalis.

    OpenAIRE

    Hinode, D; Hayashi, H.; Nakamura, R.

    1991-01-01

    Three types of caseinolytic proteases (Pase-A, Pase-B, and Pase-C) were isolated and purified from culture supernatants of Porphyromonas gingivalis 381 by the combined procedures of acetone precipitation, gel filtration, solubilization with octylthioglucoside followed by affinity chromatography on arginine-Sepharose 4B, high-performance liquid chromatography (HPLC) on Biofine IEC-DEAE, and HPLC on TSK-G4000SW. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Pase-A and -B showed ...

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

    DEFF Research Database (Denmark)

    Borch, Tanja Skuldbøl; Pedersen, Morten Løbner; Bendtzen, Klaus; Holmstrup, Palle; Nielsen, Claus Henrik

    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 (...

  15. Gingipain-dependent interactions with the host are important for survival of Porphyromonas gingivalis

    OpenAIRE

    Sheets, Shaun M.; Robles-Price, Antonette G.; McKenzie, Rachelle M. E.; Casiano, Carlos A.; Fletcher, Hansel M

    2008-01-01

    Porphyromonas gingivalis, a major periodontal pathogen, must acquire nutrients from host derived substrates, overcome oxidative stress and subvert the immune system. These activities can be coordinated via the gingipains which represent the most significant virulence factor produced by this organism. In the context of our contribution to this field, we will review the current understanding of gingipain biogenesis, glycosylation, and regulation, as well as discuss their role in oxidative stres...

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

    OpenAIRE

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

    1997-01-01

    A radioligand assay was designed to detect and compare specific hemin binding by the periodontal anaerobic black-pigmenting bacteria (BPB) Porphyromonas gingivalis and Prevotella intermedia. The assay included physiological concentrations of the hemin-binding protein rabbit serum albumin (RSA) to prevent self-aggregation and nonspecific interaction of hemin with cellular components. Under these conditions, heme-starved P. intermedia cells (two strains) expressed a single binding site species ...

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

    Directory of Open Access Journals (Sweden)

    Garcia Carlos A

    2009-05-01

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

  18. 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.

  19. The Porphyromonas gingivalis ferric uptake regulator orthologue does not regulate iron homeostasis

    Science.gov (United States)

    Butler, Catherine; Mitchell, Helen; Dashper, Stuart; Reynolds, Eric

    2015-01-01

    Porphyromonas gingivalis is a Gram-negative anaerobic bacterium that has an absolute requirement for iron which it transports from the host as heme and/or Fe2 +. Iron transport must be regulated to prevent toxic effects from excess metal in the cell. P. gingivalis has one ferric uptake regulator (Fur) orthologue encoded in its genome called Har, which would be expected to regulate the transport and usage of iron within this bacterium. As a gene regulator, inactivation of Har should result in changes in gene expression of several genes compared to the wild-type. This dataset (GEO accession number GSE37099) provides information on expression levels of genes in P. gingivalis in the absence of Har. Surprisingly, these genes do not relate to iron homeostasis.

  20. The Porphyromonas gingivalis ferric uptake regulator orthologue does not regulate iron homeostasis

    Directory of Open Access Journals (Sweden)

    Catherine Butler

    2015-09-01

    Full Text Available Porphyromonas gingivalis is a Gram-negative anaerobic bacterium that has an absolute requirement for iron which it transports from the host as heme and/or Fe2+. Iron transport must be regulated to prevent toxic effects from excess metal in the cell. P. gingivalis has one ferric uptake regulator (Fur orthologue encoded in its genome called Har, which would be expected to regulate the transport and usage of iron within this bacterium. As a gene regulator, inactivation of Har should result in changes in gene expression of several genes compared to the wild-type. This dataset (GEO accession number GSE37099 provides information on expression levels of genes in P. gingivalis in the absence of Har. Surprisingly, these genes do not relate to iron homeostasis.

  1. 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.

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

    Science.gov (United States)

    Wayakanon, Kornchanok; Thornhill, Martin H; Douglas, C W Ian; Lewis, Andrew L; Warren, Nicholas J; Pinnock, Abigail; Armes, Steven P; Battaglia, Giuseppe; Murdoch, Craig

    2013-11-01

    The gram-negative anaerobe Porphyromonas gingivalis colonizes the gingival crevice and is etiologically associated with periodontal disease that can lead to alveolar bone damage and resorption, promoting tooth loss. Although susceptible to antibiotics, P. gingivalis can evade antibiotic killing by residing within gingival keratinocytes. This provides a reservoir of organisms that may recolonize the gingival crevice once antibiotic therapy is complete. Polymersomes are nanosized amphiphilic block copolymer vesicles that can encapsulate drugs. Cells internalize polymersomes by endocytosis into early endosomes, where they are disassembled by the low pH, causing intracellular release of their drug load. In this study, polymersomes were used as vehicles to deliver antibiotics in an attempt to kill intracellular P. gingivalis within monolayers of keratinocytes and organotypic oral mucosal models. Polymersome-encapsulated metronidazole or doxycycline, free metronidazole, or doxycycline, or polymersomes alone as controls, were used, and the number of surviving intracellular P. gingivalis was quantified after host cell lysis. Polymersome-encapsulated metronidazole or doxycycline significantly (P<0.05) reduced the number of intracellular P. gingivalis in both monolayer and organotypic cultures compared to free antibiotic or polymersome alone controls. Polymersomes are effective delivery vehicles for antibiotics that do not normally gain entry to host cells. This approach could be used to treat recurrent periodontitis or other diseases caused by intracellular-dwelling organisms. PMID:23921377

  3. Arg-Gingipain A DNA Vaccine Induces Protective Immunity against Infection by Porphyromonas gingivalis in a Murine Model

    OpenAIRE

    Yonezawa, Hideo; Ishihara, Kazuyuki; Okuda, Katsuji

    2001-01-01

    Arginine-specific cysteine proteinases (RgpA and RgpB) produced by the periodontal pathogen Porphyromonas gingivalis are suspected virulence factors and are involved in interrupting host defense mechanisms as well as in penetrating and destroying periodontal connective tissues. To induce a protective immune response against P. gingivalis, we constructed an rgpA DNA vaccine. BALB/c mice were immunized intradermally by Gene Gun with plasmid DNA carrying rgpA. Antibody responses against P. gingi...

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

    OpenAIRE

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

    2008-01-01

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

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

    OpenAIRE

    Gonçalves, Reginaldo B.; Leshem, Onir; Bernards, Karen; Webb, John R; Stashenko, Philip P.; Campos-Neto, Antonio

    2006-01-01

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

  6. HmuY Haemophore and Gingipain Proteases Constitute a Unique Syntrophic System of Haem Acquisition by Porphyromonas gingivalis

    OpenAIRE

    Smalley, John W; Byrne, Dominic P.; Birss, Andrew J; Wojtowicz, Halina; Sroka, Aneta; Potempa, Jan; Olczak, Teresa

    2011-01-01

    Haem (iron protoporphyrin IX) is both an essential growth factor and virulence regulator for the periodontal pathogen Porphyromonas gingivalis, which acquires it mainly from haemoglobin via the sequential actions of the R- and K-specific gingipain proteases. The haem-binding lipoprotein haemophore HmuY and its cognate receptor HmuR of P. gingivalis, are responsible for capture and internalisation of haem. This study examined the role of the HmuY in acquisition of haem from haemoglobin and the...

  7. A Dual Role for P2X7 Receptor during Porphyromonas gingivalis Infection.

    Science.gov (United States)

    Ramos-Junior, E S; Morandini, A C; Almeida-da-Silva, C L C; Franco, E J; Potempa, J; Nguyen, K A; Oliveira, A C; Zamboni, D S; Ojcius, D M; Scharfstein, J; Coutinho-Silva, R

    2015-09-01

    Emerging evidence suggests a role for purinergic signaling in the activation of multiprotein intracellular complexes called inflammasomes, which control the release of potent inflammatory cytokines, such as interleukin (IL) -1? and -18. Porphyromonas gingivalis is intimately associated with periodontitis and is currently considered one of the pathogens that can subvert the immune system by limiting the activation of the NLRP3 inflammasome. We recently showed that P. gingivalis can dampen eATP-induced IL-1? secretion by means of its fimbriae in a purinergic P2X7 receptor-dependent manner. Here, we further explore the role of this purinergic receptor during eATP-induced IL-1? processing and secretion by P. gingivalis-infected macrophages. We found that NLRP3 was necessary for eATP-induced IL-1? secretion as well as for caspase 1 activation irrespective of P. gingivalis fimbriae. Additionally, although the secretion of IL-1? from P. gingivalis-infected macrophages was dependent on NLRP3, its adaptor protein ASC, or caspase 1, the cleavage of intracellular pro-IL-1? to the mature form was found to occur independently of NLRP3, its adaptor protein ASC, or caspase 1. Our in vitro findings revealed that P2X7 receptor has a dual role, being critical not only for eATP-induced IL-1? secretion but also for intracellular pro-IL-1? processing. These results were relevant in vivo since P2X7 receptor expression was upregulated in a P. gingivalis oral infection model, and reduced IFN-? and IL-17 were detected in draining lymph node cells from P2rx7(-/-) mice. Furthermore, we demonstrated that P2X7 receptor and NLRP3 transcription were modulated in human chronic periodontitis. Overall, we conclude that the P2X7 receptor has a role in periodontal immunopathogenesis and suggest that targeting of the P2X7/NLRP3 pathway should be considered in future therapeutic interventions in periodontitis. PMID:26152185

  8. Discrete protein determinant directs the species-specific adherence of Porphyromonas gingivalis to oral streptococci.

    Science.gov (United States)

    Demuth, D R; Irvine, D C; Costerton, J W; Cook, G S; Lamont, R J

    2001-09-01

    For pathogens to survive in the human oral cavity, they must identify a suitable niche in the complex multispecies biofilm that exists on oral tissues. The periodontal pathogen Porphyromonas gingivalis adheres to Streptococcus gordonii by interacting with a specific region of the streptococcal SspB polypeptide, designated BAR. However, it does not adhere to Streptococcus mutans, which expresses SpaP, a highly conserved homolog of SspB. Comparison of the predicted secondary structure of BAR with the corresponding region of SpaP suggested that the substitution of Asn for Gly1182 and Val for Pro1185 in SspB may confer a unique local structure that is not conserved in SpaP. A synthetic peptide of 26 amino acids that encompassed residues 1167 to 1193 of SspB promoted avid adherence of P. gingivalis, whereas a peptide derived from the region corresponding to BAR in SpaP was inactive. Substitution of Gly1182 and Pro1185 for Asn1182 and Val1185 in SspB by site-specific mutation generated proteins that were predicted to assume an SpaP-like secondary structure, and the purified proteins did not promote P. gingivalis adherence. Furthermore, Enterococcus faecalis strains expressing the site-specific mutants did not support adherence of P. gingivalis cells. In contrast, P. gingivalis adhered efficiently to E. faecalis strains expressing intact SspB or SspB-SpaP chimeric proteins containing BAR. These results suggest that a region of SspB consisting of 26 amino acids is sufficient to mediate the adherence of P. gingivalis to S. gordonii and that the species specificity of adherence arises from its interaction with a discrete structural determinant of SspB that is not conserved in SpaP. PMID:11500450

  9. Porphyromonas gingivalis Ferrous Iron Transporter FeoB1 Influences Sensitivity to Oxidative Stress ?

    Science.gov (United States)

    Anaya-Bergman, Cecilia; He, Jia; Jones, Kevin; Miyazaki, Hiroshi; Yeudall, Andrew; Lewis, Janina P.

    2010-01-01

    Porphyromonas gingivalis FeoB1 is a ferrous iron transporter. Analysis of parental and feoB1-deficient strains of the periodontal pathogen revealed that the feoB1-deficient mutant strain had an increased ability to survive oxidative stress. Specifically, survival of the mutant strain was increased 33% with exposure to peroxide and 5% with exposure to atmospheric oxygen compared to the parental strain. Interestingly, the ability to survive intracellularly also increased fivefold in the case of the feoB1-deficient mutant. Our data suggest that although the FeoB1 protein is required for ferrous iron acquisition in P. gingivalis, it also has an adverse effect on survival of the bacterium under oxidative stress conditions. Finally, we show that feoB1 expression is not iron dependent and is dramatically reduced in the presence of host cells, consistent with the observed deleterious role it plays in bacterial survival. PMID:19917713

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

    Science.gov (United States)

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

    2012-01-01

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

  11. Porphyromonas gingivalis fimbriae dampen P2X7-dependent IL-1? secretion

    OpenAIRE

    Morandini, Ana Carolina; Ramos-Junior, Erivan S.; Potempa, Jan; Nguyen, Ky-Anh; Oliveira, Ana Carolina; Bellio, Maria; Ojcius, David M; Scharfstein, Julio; Coutinho-Silva, Robson

    2014-01-01

    Porphyromonas gingivalis is a major contributor to the pathogenesis of periodontitis, an infection-driven inflammatory disease that leads to bone destruction. This pathogen stimulates pro-IL-1? synthesis but not mature IL-1? secretion, unless the P2X7 receptor is activated by extracellular ATP. Here, we investigated the role of Pg fimbriae in eATP-induced IL-1? release. Bone marrow derived macrophages (BMDMs) from wild type (WT) or P2X7-deficient mice were infected with Pg (...

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

    International Nuclear Information System (INIS)

    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

  13. 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.

  14. The role of phagocytosis, oxidative burst and neutrophil extracellular traps in the interaction between neutrophils and the periodontal pathogen Porphyromonas gingivalis.

    Science.gov (United States)

    Jayaprakash, K; Demirel, I; Khalaf, H; Bengtsson, T

    2015-10-01

    Neutrophils are regarded as the sentinel cells of innate immunity and are found in abundance within the gingival crevice. Discovery of neutrophil extracellular traps (NETs) within the gingival pockets prompted us to probe the nature of the interactions of neutrophils with the prominent periopathogen Porphyromonas gingivalis. Some of the noted virulence factors of this Gram-negative anaerobe are gingipains: arginine gingipains (RgpA/B) and lysine gingipain (Kgp). The aim of this study was to evaluate the role of gingipains in phagocytosis, formation of reactive oxygen species, NETs and CXCL8 modulation by using wild-type strains and isogenic gingipain mutants. Confocal imaging showed that gingipain mutants K1A (Kgp) and E8 (RgpA/B) induced extracellular traps in neutrophils, whereas ATCC33277 and W50 were phagocytosed. The viability of both ATCC33277 and W50 dwindled as the result of phagocytosis and could be salvaged by cytochalasin D, and the bacteria released high levels of lipopolysaccharide in the culture supernatant. Porphyromonas gingivalis induced reactive oxygen species and CXCL8 with the most prominent effect being that of the wild-type strain ATCC33277, whereas the other wild-type strain W50 was less effective. Quantitative real-time polymerase chain reaction revealed a significant CXCL8 expression by E8. All the tested P. gingivalis strains increased cytosolic free calcium. In conclusion, phagocytosis is the primary neutrophil response to P. gingivalis, although NETs could play an accessory role in infection control. Although gingipains do not seem to directly regulate phagocytosis, NETs or oxidative burst in neutrophils, their proteolytic properties could modulate the subsequent outcomes such as nutrition acquisition and survival by the bacteria. PMID:25869817

  15. 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.

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

    Science.gov (United States)

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

    2002-06-01

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

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

    Directory of Open Access Journals (Sweden)

    Amir E. Zeituni

    2010-12-01

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

  18. 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

  19. 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.

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

    Directory of Open Access Journals (Sweden)

    Antonio Díaz Caballero

    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 artículo 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.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 review 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.

  1. Identification of amino acid residues involved in hemin binding in Porphyromonas gingivalis hemagglutinin 2.

    Science.gov (United States)

    Yang, Q B; Yu, F Y; Sun, L; Zhang, Q X; Lin, M; Geng, X Y; Sun, X N; Li, J L; Liu, Y

    2015-10-01

    Porphyromonas gingivalis (P. gingivalis) is a major etiological agent in the development and progression of chronic periodontitis. It produces cysteine proteases (gingipains), including a lysine-specific gingipain and two arginine-specific gingipains. Heme binding and uptake are fundamental to the growth and virulence of P. gingivalis. The recombinant hemagglutinin 2 domain (rHA2) of gingipain binds hemin with high affinity. The aim of the present work was to identify the key residues involved in its hemin-binding activity. A functional rHA2 was expressed and bound to hemin-agarose, and then digested with endopeptidases. The peptides bound to hemin-agarose were identified by mass spectrometry and the amino acids were assessed by mutation and peptide binding inhibition analysis. The DHYAVMISK sequence was identified in peptides derived from both Asp-N and Lys-C endopeptidase digestions of rHA2. A monoclonal antibody, mAb QB, was produced and its epitope was associated with the DGFPGDHYAVMISK peptide within the HA2 domain. Hemin was shown to competitively inhibit the immunoreactivity of rHA2 or the peptide to mAb QB. The peptide DHYAVMISK inhibited hemin-binding activity; although, this inhibition was not seen when the peptide contained the H1001E mutation (DEYAVMISK). Based on these results, we propose that residue His1001 is involved in the hemin-binding mechanism of the P. gingivalis rHA2 and the peptide containing this residue, DHYAVMISK, may be an inhibitor of hemin binding. PMID:25833325

  2. Intraspecies Variability Affects Heterotypic Biofilms of Porphyromonas gingivalis and Prevotella intermedia: Evidences of Strain-Dependence Biofilm Modulation by Physical Contact and by Released Soluble Factors

    Science.gov (United States)

    Barbosa, Graziela Murta; Colombo, Andrea Vieira; Rodrigues, Paulo Henrique; Simionato, Maria Regina Lorenzetti

    2015-01-01

    It is well known that strain and virulence diversity exist within the population structure of Porphyromonas gingivalis. In the present study we investigate intra- and inter-species variability in biofilm formation of Porphyromonas gingivalis and partners Prevotella intermedia and Prevotella nigrescens. All strains tested showed similar hydrophobicity, except for P. gingivalis W83 which has roughly half of the hydrophobicity of P. gingivalis ATCC33277. An intraspecies variability in coaggregation of P. gingivalis with P. intermedia was also found. The association P. gingivalis W83/P. intermedia 17 produced the thickest biofilm and strain 17 was prevalent. In a two-compartment system P. gingivalis W83 stimulates an increase in biomass of strain 17 and the latter did not stimulate the growth of P. gingivalis W83. In addition, P. gingivalis W83 also stimulates the growth of P. intermedia ATCC25611 although strain W83 was prevalent in the association with P. intermedia ATCC25611. P. gingivalis ATCC33277 was prevalent in both associations with P. intermedia and both strains of P. intermedia stimulate the growth of P. gingivalis ATCC33277. FISH images also showed variability in biofilm structure. Thus, the outcome of the association P. gingivalis/P. intermedia seems to be strain-dependent, and both soluble factors and physical contact are relevant. The association P. gingivalis-P. nigrescens ATCC33563 produced larger biomass than each monotypic biofilm, and P. gingivalis was favored in consortia, while no differences were found in the two-compartment system. Therefore, in consortia P. gingivalis-P. nigrescens physical contact seems to favor P. gingivalis growth. The intraspecies variability found in our study suggests strain-dependence in ability of microorganisms to recognize molecules in other bacteria which may further elucidate the dysbiosis event during periodontitis development giving additional explanation for periodontal bacteria, such as P. gingivalis and P. intermedia, among others, to persist and establish chronic infections in the host. PMID:26406499

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

    International Nuclear Information System (INIS)

    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 1H 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.

  4. LuxS Involvement in the Regulation of Genes Coding for Hemin and Iron Acquisition Systems in Porphyromonas gingivalis

    OpenAIRE

    James, Chloe E; Hasegawa, Yoshiaki; Park, Yoonsuk; Yeung, Vincent; Tribble, Gena D.; Kuboniwa, Masae; Demuth, Donald R.; Lamont, Richard J.

    2006-01-01

    The periodontal pathogen Porphyromonas gingivalis employs a variety of mechanisms for the uptake of hemin and inorganic iron. Previous work demonstrated that hemin uptake in P. gingivalis may be controlled by LuxS-mediated signaling. In the present study, the expression of genes involved in hemin and iron uptake was determined in parent and luxS mutant strains by quantitative real-time reverse transcription-PCR. Compared to the parental strain, the luxS mutant showed reduced levels of transcr...

  5. Identification of gingipain-specific I-Ab-restricted CD4+ T cells following mucosal colonization with Porphyromonas gingivalis in C57BL/6 mice

    OpenAIRE

    Bittner-Eddy, PD; Fischer, LA; Costalonga, M

    2013-01-01

    Chronic periodontitis is associated with Porphyromonas gingivalis infection. Although virulence factors of P. gingivalis are hypothesized to contribute to the pathogenesis of periodontitis, it is unclear whether the local CD4+ T-cell-mediated response they elicit prevents or contributes to periodontal bone destruction. We hypothesize that major histocompatibility complex class II I-Ab-binding peptides existing in Kgp and RgpA are presented to CD4+ T cells during P. gingivalis oral colonizatio...

  6. 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...

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

    Science.gov (United States)

    Wang, Yu-Hsiung; Nemati, Reza; Anstadt, Emily; Liu, Yaling; Son, Young; Zhu, Qiang; Yao, Xudong; Clark, Robert B; Rowe, David W; Nichols, Frank C

    2015-12-01

    Porphyromonas gingivalis is a periodontal pathogen strongly associated with loss of attachment and supporting bone for teeth. We have previously shown that the total lipid extract of P. gingivalis inhibits osteoblast differentiation through engagement of Toll-like receptor 2 (TLR2) and that serine dipeptide lipids of P. gingivalis engage both mouse and human TLR2. The purpose of the present investigation was to determine whether these serine lipids inhibit osteoblast differentiation in vitro and in vivo and whether TLR2 engagement is involved. Osteoblasts were obtained from calvaria of wild type or TLR2 knockout mouse pups that also express the Col2.3GFP transgene. Two classes of serine dipeptide lipids, termed Lipid 654 and Lipid 430, were tested. Osteoblast differentiation was monitored by cell GFP fluorescence and osteoblast gene expression and osteoblast function was monitored as von Kossa stained mineral deposits. Osteoblast differentiation and function were evaluated in calvarial cell cultures maintained for 21days. Lipid 654 significantly inhibited GFP expression, osteoblast gene expression and mineral nodule formation and this inhibition was dependent on TLR2 engagement. Lipid 430 also significantly inhibited GFP expression, osteoblast gene expression and mineral nodule formation but these effects were only partially attributed to engagement of TLR2. More importantly, Lipid 430 stimulated TNF-? and RANKL gene expression in wild type cells but not in TLR2 knockout cells. Finally, osteoblast cultures were observed to hydrolyze Lipid 654 to Lipid 430 and this likely occurs through elevated PLA2 activity in the cultured cells. In conclusion, our results show that serine dipeptide lipids of P. gingivalis inhibit osteoblast differentiation and function at least in part through engagement of TLR2. The Lipid 430 serine class also increased the expression of genes that could increase osteoclast activity. We conclude that Lipid 654 and Lipid 430 have the potential to promote TLR2-dependent bone loss as is reported in experimental periodontitis following oral infection with P. gingivalis. These results also support the conclusion that serine dipeptide lipids are involved in alveolar bone loss in chronic periodontitis. PMID:26409254

  8. 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.

  9. 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.

  10. Role of the Porphyromonas gingivalis ECF sigma factor, SigH

    Science.gov (United States)

    Yanamandra, Sai S.; Sarrafee, Sara S.; Anaya-Bergman, Cecilia; Jones, Kevin; Lewis, Janina P.

    2012-01-01

    Little is known about the regulatory mechanisms that allow Porphyromonas gingivalis to survive in the oral cavity. Here we characterize the sigma factor SigH, one of six extracytoplasmic (ECF) sigma (?) factors encoded in the P. gingivalis genome. Our results indicate that sigH expression is upregulated by exposure to molecular oxygen, suggesting that sigH plays a role in adaptation of P. gingivalis to oxygen. Furthermore, several genes involved in oxidative stress protection, such as sod, trx, tpx, ftn, feoB2 and the hemin uptake hmu locus, are downregulated in mutant deficient in SigH designated as V2948. ECF ? consensus sequences were identified upstream of the transcriptional start sites of these genes, consistent with the SigH-dependent regulation of these genes. Growth of V2948 was inhibited in the presence of 6% oxygen when compared to the wild-type W83 strain, while in anaerobic conditions both strains were able to grow. In addition, reduced growth of V2948 was observed in the presence of peroxide and thiol-oxidizing reagent, diamide when compared to the W83 strain. The SigH-deficient strain V2948 also exhibited reduced hemin uptake, consistent with the observed reduced expression of genes involved in hemin uptake. Finally, survival of V2948 was reduced in the presence of host cells compared to the wild-type W83 strain. Collectively, our studies demonstrate that SigH is a positive regulator of gene expression required for survival of the bacterium in the presence of oxygen and oxidative stress, hemin uptake, and virulence. PMID:22520389

  11. Regulation of protease-activated receptor-2 expression in gingival fibroblasts and Jurkat T cells by Porphyromonas gingivalis

    OpenAIRE

    Belibasakis, G.N.; Bostanci, N; Reddi, D

    2010-01-01

    Periodontal disease destroys the tooth-supporting tissues as a result of chronic inflammation elicited by bacterial accumulation on tooth surfaces. Porphyromonas gingivalis is a major periodontal pathogen, with a significant capacity to perturb connective tissue homeostasis and immune responses in the periodontium, attributed to its virulence factors, including a group of secreted cysteine proteases (gingipains). PAR-2 (protease-activated receptor-2) is a G-protein-coupled receptor activated ...

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

    DEFF Research Database (Denmark)

    Veillard, Florian; Sztukowska, Maryta; Mizgalska, Danuta; Ksiazek, Miros?aw; Houston, John; Potempa, Barbara; Enghild, Jan Johannes; Thøgersen, Ida B; Gomis-Rüth, F Xavier; Nguyen, Ky-Anh; Potempa, Jan

    2013-01-01

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

  13. 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; Bendtzen, K; Nielsen, Claus Henrik

    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 inte...

  14. Hemin levels in culture medium of Porphyromonas (Bacteroides) gingivalis regulate both hemin binding and trypsinlike protease production.

    OpenAIRE

    Carman, R.J.; Ramakrishnan, M D; Harper, F. H.

    1990-01-01

    Washed cells and Sarkosyl-insoluble outer membrane preparations of the black-pigmented bacteroides Porphyromonas gingivalis W50 bound hemin. The amount of hemin removed from a buffered solution by both cells and outer membranes was significantly larger if bacteria had been grown in broths supplemented with 5 mg of hemin per liter rather than none. Conversely, cells grown without supplemental hemin bound relatively little. However, all preparations bound some hemin. In addition, hemin regulate...

  15. Unique Structure and Stability of HmuY, a Novel Heme-Binding Protein of Porphyromonas gingivalis

    OpenAIRE

    2009-01-01

    Infection, survival, and proliferation of pathogenic bacteria in humans depend on their capacity to impair host responses and acquire nutrients in a hostile environment. Among such nutrients is heme, a co-factor for oxygen storage, electron transport, photosynthesis, and redox biochemistry, which is indispensable for life. Porphyromonas gingivalis is the major human bacterial pathogen responsible for severe periodontitis. It recruits heme through HmuY, which sequesters heme from host carriers...

  16. 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

  17. Determination of the antibacterial activity of simvastatin against periodontal pathogens, Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans: An in vitro study

    Directory of Open Access Journals (Sweden)

    Shilpa Emani

    2014-01-01

    Full Text Available Context and Objective: Statin treatment, apart from its hypolipidemic action has proven its antimicrobial activity by improving the survival rate of patients with severe systemic bacterial infections. Periodontitis is an inflammatory disorder of tooth supporting structures caused by a group of specific microorganisms. The objective of the present study was to determine the antimicrobial activity of pure simvastatin drug against the primary periodontal pathogens. Materials and Methods: Minimum inhibitory concentration (MIC was determined against Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans using serial dilution method. Results: MIC of simvastatin against P. gingivalis was 2 ?g/ml and A. actinomycetemcomitans was found to be <1 ?g/ml which requires further dilutions to determine the exact value. Conclusions: Data suggests a potent antimicrobial activity of simvastatin against both A. actinomycetemcomitans and P gingivalis. Hence simvastatin can be prescribed as a dual action drug in patients with both hyperlipidemia and periodontal disease.

  18. Porphyromonas gingivalis HSP60 peptides have distinct roles in the development of atherosclerosis.

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    Jeong, Euikyong; Kim, Koanhoi; Kim, June Hong; Cha, Go Sim; Kim, Sung-Jo; Kang, Ho Sung; Choi, Jeomil

    2015-02-01

    Different epitope peptides of bacterial heat shock proteins may function as effector or regulatory molecules in autoimmune responses in infection-triggered atherosclerosis. We investigated the mechanisms for the distinct roles of two epitope peptides from Porphyromonas gingivalis heat shock protein 60 (HSP60) in atherogenesis with regard to peptide-specific T cell polarization relevant to (1) phenotype and cytokine profiles, (2) expression of transcription factors, and (3) role of antigen presenting dendritic cell subsets.Apolipoprotein E-knockout (ApoE KO) mice were immunized with peptide 14 or peptide 19 from P. gingivalis HSP60 prior to induction of atherosclerosis by infection with P. gingivalis plus a Western diet. Significant reductions in plaque/lipid droplet area and plasma cholesterol levels were observed in mice immunized with peptide 14, whereas the opposite phenomenon was evident in mice immunized with peptide 19. CD4+ T-cells polarized to the regulatory T-cell type in peptide 14-immunized group, whereas they polarized to the Th1 cells in peptide 19-immunized group; this observation was supported by the cytokine profiles characteristic to each T-cell phenotype.Significantly higher expression of Nr4a1 and Nr4a2 mRNA, transcriptional factors for regulatory T-cell type, were observed in peptide 14-immunized group. In contrast, the expression level of IFN-? and T-bet mRNA, signaling molecules for Th1 cells, was higher in peptide 19-immunized group than in PBS-immunized group.In non-immunized wild mice, BMDC-derived CD11c+ dendritic cells have shown to stimulate Tregs significantly in antigen-nonspecific manner. However, each peptide antigen demonstrated a unique mode of preferential adoption of dendritic cell subsets.In conclusion, peptide 14 or peptide 19 from P. gingivalis HSP60, respectively, may play either an anti- or pro-atherogenic role in the ApoE KO mouse model of infection-triggered atherosclerosis through distinct mechanisms operating in the polarization of T cells. PMID:25457882

  19. 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

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    J Díaz Zúñiga

    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 periodontales a través de los diversos factores de virulencia que expresan. Sobre la base de estos factores de virulencia, distintos genotipos y serotipos bacterianos se han 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.Periodontitis represents a heterogenic group of periodontal infections elicited by bacteria residing at the subgingival biofilm. Although this biofilm is constituted by a broad variety of bacterial species, only a limited number has been associated with the periodontitis aetiology, among them Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans. Both P. gingivalis and A. actinomycetemcomitans express a number of virulence factors that contribute to direct 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.

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

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    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.

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

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    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

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

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    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.

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

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    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.

  4. Rosiglitazone impedes Porphyromonas gingivalis-accelerated atherosclerosis by downregulating the TLR/NF-?B signaling pathway in atherosclerotic mice.

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    Pan, Shengbo; Lei, Lang; Chen, Shuai; Li, Houxuan; Yan, Fuhua

    2014-12-01

    Porphyromonas gingivalis,a predominant periodontal pathogen, is known to accelerate atherosclerosis in hyperlipidemic animals via aberrant inflammatory responses. Peroxisome proliferator-activated receptor gamma (PPAR?) agonists have been reported to exert anti-inflammatory effects in vitro. The purpose of the present study was to investigate the potential protective role of the PPAR? agonist rosiglitazone in pathogen accelerated atherosclerosis in an apolipoprotein E-deficient (ApoE-/-) mouse model. ApoE-/- mice were inoculated intravenously with live P. gingivalis (strain 33277) or the buffer vehicle and treated with rosiglitazone or saline over a 10-week period. Their atherosclerotic status in aortic artery was assessed through histomorphometric analysis, inflammatory agent and lipid profiles in blood was determined by ELISA, and levels of relevant cytokines and Toll-like receptors (TLRs) in aortic tissues were evaluated using immunohistochemistry and quantitative PCR. P. gingivalis inoculation was associated with increased atherosclerotic plaque formation in the aorta and higher levels of serum pro-inflammatory cytokines (tumor necrosis factor-?, monocyte chemotactic protein-1 and interleukin-1?), but the serum lipid profile was not affected by P. gingivalis infection. Levels of tumor necrosis factor-?, monocyte chemotactic protein-1 intercellular cell adhesion molecule-1 and TLRs were higher in the aortic tissues of mice exposed to P. gingivalis, and activation of nuclear factor-?B was also observed. In both P. gingivalis-treated and -untreated ApoE-/- mice, rosiglitazone treatment was associated with less atherosclerotic plaque formation; lower serum inflammatory cytokines, total cholesterol, and low density lipoprotein cholesterol; higher levels of PPAR?, lower amounts of TLR2/4 and downregulated nuclear factor-?B activity in aortic tissues. These findings suggest that rosiglitazone mitigates or prevents P. gingivalis-accelerated atherosclerosis by inhibiting the inflammatory response via downregulation of the TLR/ nuclear factor-?B signaling pathway. PMID:25445963

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

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    Zohreh Tamanai-Shacoori

    2009-06-01

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

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

    Scientific Electronic Library Online (English)

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

    2009-06-01

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

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

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    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.

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

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    Dietrich Elizabeth A

    2008-03-01

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

  9. 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

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    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

  10. Localization of a Porphyromonas gingivalis 26-kilodalton heat-modifiable, hemin-regulated surface protein which translocates across the outer membrane.

    OpenAIRE

    Bramanti, T E; Holt, S.C.

    1992-01-01

    We recently identified a 26-kDa hemin-repressible outer membrane protein (Omp26) expressed by the periodontal pathogen Porphyromonas gingivalis. We report the localization of Omp26, which may function as a component of a hemin transport system in P. gingivalis. Under hemin-deprived conditions, P. gingivalis expressed Omp26, which was then lost from the surface after a shift back into hemin-rich conditions. Experiments with 125I labeling of surface proteins to examine the kinetics of mobilizat...

  11. 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.

  12. LPS from Porphyromonas gingivalis increases the sensitivity of contractile response mediated by endothelin-B (ET(B)) receptors in cultured endothelium-intact rat coronary arteries

    DEFF Research Database (Denmark)

    Ghorbani, Bahareh; Holmstrup, Palle

    2010-01-01

    The purpose of our study was to examine if lipopolysaccharide (LPS) from Porphyromonas gingivalis (P.g.) modifies the vasomotor responses to Endothelin-1 (ET-1) and Sarafotoxin 6c (S6c) in rat coronary arteries. The arteries were studied directly or following organ culture for 24 h in absence and presence of 2.5EU/ml LPS. The contractile responses of coronary arteries were investigated by using the selective ETB receptor agonist S6c (1 pM-0.3 ?M) and ET-1 (1 pM-0.3 ?M). The functional studies demonstrated an augmented contractile response only to S6c in isolated rat coronary arteries after organ culture (with or without LPS). These contractile responses by S6c were blocked by the selective ETB receptor antagonist BQ788 in both vessel groups. The augmented contractile response to S6c was supported by immunohistochemistry, where a significant increase in fluorescence intensity for ETB receptors in smooth muscle cells was observed after organ culture. The presence of LPS in the culture medium significantly increased the sensitivity of endothelium-intact coronary artery to S6c as compared to endothelium-denuded segments. Our results showed a significant increase in both ETB receptor protein levels and S6c-induced maximal contraction in coronary arteries upon 24 h of organ culture, which was further sensitized by LPS.

  13. 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

  14. 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.

  15. 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....

  16. 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

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

    Science.gov (United States)

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

    1994-03-30

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

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

    OpenAIRE

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

    2010-01-01

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

  19. Characterization of hemin-binding protein 35 (HBP35) in Porphyromonas gingivalis: its cellular distribution, thioredoxin activity and role in heme utilization

    OpenAIRE

    Abiko Yoshimitsu; Naito Mariko; Sato Keiko; Chen Yu-Yen; Peng Benjamin; Yukitake Hideharu; Shiroza Teruaki; Shibata Yasuko; Shoji Mikio; Reynolds Eric C; Nakayama Koji

    2010-01-01

    Abstract Background The periodontal pathogen Porphyromonas gingivalis is an obligate anaerobe that requires heme for growth. To understand its heme acquisition mechanism, we focused on a hemin-binding protein (HBP35 protein), possessing one thioredoxin-like motif and a conserved C-terminal domain, which are proposed to be involved in redox regulation and cell surface attachment, respectively. Results We observed that the hbp35 gene was transcribed as a 1.1-kb mRNA with subsequent translation ...

  20. Dual Action of Myricetin on Porphyromonas gingivalis and the Inflammatory Response of Host Cells: A Promising Therapeutic Molecule for Periodontal Diseases.

    Science.gov (United States)

    Grenier, Daniel; Chen, Huangqin; Ben Lagha, Amel; Fournier-Larente, Jade; Morin, Marie-Pierre

    2015-01-01

    Periodontitis that affects the underlying structures of the periodontium, including the alveolar bone, is a multifactorial disease, whose etiology involves interactions between specific bacterial species of the subgingival biofilm and the host immune components. In the present study, we investigated the effects of myricetin, a flavonol largely distributed in fruits and vegetables, on growth and virulence properties of Porphyromonas gingivalis as well as on the P. gingivalis-induced inflammatory response in host cells. Minimal inhibitory concentration values of myricetin against P. gingivalis were in the range of 62.5 to 125 ?g/ml. The iron-chelating activity of myricetin may contribute to the antibacterial activity of this flavonol. Myricetin was found to attenuate the virulence of P. gingivalis by reducing the expression of genes coding for important virulence factors, including proteinases (rgpA, rgpB, and kgp) and adhesins (fimA, hagA, and hagB). Myricetin dose-dependently prevented NF-?B activation in a monocyte model. Moreover, it inhibited the secretion of IL-6, IL-8 and MMP-3 by P. gingivalis-stimulated gingival fibroblasts. In conclusion, our study brought clear evidence that the flavonol myricetin exhibits a dual action on the periodontopathogenic bacterium P. gingivalis and the inflammatory response of host cells. Therefore, myricetin holds promise as a therapeutic agent for the treatment/prevention of periodontitis. PMID:26121135

  1. Effects of the antimicrobial peptide cathelicidin (LL-37) on immortalized gingival fibroblasts infected with Porphyromonas gingivalis and irradiated with 625-nm LED light.

    Science.gov (United States)

    Kim, JiSun; Kim, SangWoo; Lim, WonBong; Choi, HongRan; Kim, OkJoon

    2015-11-01

    Porphyromonas gingivalis causes chronic inflammatory diseases (periodontal diseases) that destroy the periodontal ligament and alveolar bone. Antimicrobial peptides are crucial components of the host defense response required to maintain cellular homeostasis during microbial invasion. Because light-emitting diode (LED) irradiation influences the host defense response against bacterial infections, we investigated its effect on immortalized gingival fibroblasts (IGFs) infected with P. gingivalis. IGFs were incubated with P. gingivalis following LED irradiation at 425, 525, and 625 nm. The dark 1 group comprised noninfected, nonirradiated IGFs, and the dark 2 group comprised nonirradiated IGFs infected with P. gingivalis. These groups served as controls. Infected cells and controls were assayed for reactive oxygen species (ROS) and were subjected to RT-PCR and Western blotting analyses to determine the levels of expression of antimicrobial peptides. LED irradiation enhanced the bactericidal effects of the antimicrobial peptide LL-37 in cells infected with P. gingivalis. Irradiation at 625 nm decreased inflammatory responses involving the release of prostaglandin E2 induced by ROS in P. gingivalis-infected IGFs. LED irradiation at 625 nm induces an anti-inflammatory response that elicits the production of antimicrobial peptides, providing an efficacious method of treatment for periodontal diseases. PMID:25543295

  2. Biochemical characterization of recombinant ?-carbonic anhydrase (PgiCAb) identified in the genome of the oral pathogenic bacterium Porphyromonas gingivalis.

    Science.gov (United States)

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

    2015-06-01

    Carbonic anhydrases (CAs, EC 4.2.1.1) belonging to the ?-, ?-, ?-, ?- and ?-CAs are ubiquitous metalloenzymes present in prokaryotes and eukaryotes. CAs started to be investigated in detail only recently in pathogenic bacteria, in the search for antibiotics with a novel mechanism of action, since it has been demonstrated that in many such organisms they are essential for the life cycle of the organism. CA inhibition leads to growth impairment or growth defects in several pathogenic bacteria. The microbiota of the human oral mucosa consists of a myriad of bacterial species, Porphyromonas gingivalis being one of them and the major pathogen responsible for the development of chronic periodontitis. The genome of P. gingivalis encodes for a ?- and a ?-CAs. Recently, our group purified the recombinant ?-CA (named PgiCA) which was shown to possess a significant catalytic activity for the reaction that converts CO2 to bicarbonate and protons, with a kcat of 4.1?×?10(5?)s(-1) and a kcat/Km of 5.4?×?10(7?)M(-1?)×?s(-1). We have also investigated its inhibition profile with a range of inorganic anions such as thiocyanate, cyanide, azide, hydrogen sulfide, sulfamate and trithiocarbonate. Here, we describe the cloning, purification and kinetic parameters of the other class of CA identified in the genome of P. gingivalis, the ?-CA, named PgiCAb. This enzyme has a good catalytic activity, with a kcat of 2.8?×?10(5?)s(-1) and a kcat/Km of 1.5?×?10(7?)M(-1?)×?s(-1). PgiCAb was also inhibited by the clinically used sulfonamide acetazolamide, with an inhibition constant of 214?nM. The role of CAs as possible virulence factors of P. gingivalis is poorly understood at the moment but their good catalytic activity and the fact that they might be inhibited by a large number of compounds, which may pave the way for finding inhibitors with antibacterial activity that may elucidate these phenomena and lead to novel antibiotics. PMID:25032746

  3. A Porphyromonas gingivalis Tyrosine Phosphatase is a Multifunctional Regulator of Virulence Attributes

    OpenAIRE

    Maeda, Kazuhiko; Tribble, Gena D.; Tucker, Chelsea M.; Anaya, Cecilia; Shizukuishi, Satoshi; Lewis, Janina P.; Demuth, Donald R.; Lamont, Richard J.

    2008-01-01

    Low Molecular Weight Tyrosine Phosphatases (LMWTP) are widespread in prokaryotes; however, understanding of the signaling cascades controlled by these enzymes is still emerging. P. gingivalis, an opportunistic oral pathogen, expresses a LMWTP, Ltp1, that is differentially regulated in biofilm communities. Here we characterize the enzymatic activity of Ltp1 and, through the use of mutants that lack Ltp1 or expresses catalytically defective Ltp1, show that tyrosine phosphatase activity constrai...

  4. Gingipains from the Periodontal Pathogen Porphyromonas gingivalis Play a Significant Role in Regulation of Angiopoietin 1 and Angiopoietin 2 in Human Aortic Smooth Muscle Cells.

    Science.gov (United States)

    Zhang, Boxi; Khalaf, Hazem; Sirsjö, Allan; Bengtsson, Torbjörn

    2015-11-01

    Angiopoietin 1 (Angpt1) and angiopoietin 2 (Angpt2) are the ligands of tyrosine kinase (Tie) receptors, and they play important roles in vessel formation and the development of inflammatory diseases, such as atherosclerosis. Porphyromonas gingivalis is a Gram-negative periodontal bacterium that is thought to contribute to the progression of cardiovascular disease. The aim of this study was to investigate the role of P. gingivalis infection in the modulation of Angpt1 and Angpt2 in human aortic smooth muscle cells (AoSMCs). We exposed AoSMCs to wild-type (W50 and 381), gingipain mutant (E8 and K1A), and fimbrial mutant (DPG-3 and KRX-178) P. gingivalis strains and to different concentrations of tumor necrosis factor (TNF). The atherosclerosis risk factor TNF was used as a positive control in this study. We found that P. gingivalis (wild type, K1A, DPG3, and KRX178) and TNF upregulated the expression of Angpt2 and its transcription factor ETS1, respectively, in AoSMCs. In contrast, Angpt1 was inhibited by P. gingivalis and TNF. However, the RgpAB mutant E8 had no effect on the expression of Angpt1, Angpt2, or ETS1 in AoSMCs. The results also showed that ETS1 is critical for P. gingivalis induction of Angpt2. Exposure to Angpt2 protein enhanced the migration of AoSMCs but had no effect on proliferation. This study demonstrates that gingipains are crucial to the ability of P. gingivalis to markedly increase the expressed Angpt2/Angpt1 ratio in AoSMCs, which determines the regulatory role of angiopoietins in angiogenesis and their involvement in the development of atherosclerosis. These findings further support the association between periodontitis and cardiovascular disease. PMID:26283334

  5. GM-CSF and uPA are required for Porphyromonas gingivalis-induced alveolar bone loss in a mouse periodontitis model.

    Science.gov (United States)

    Lam, Roselind S; O'Brien-Simpson, Neil M; Hamilton, John A; Lenzo, Jason C; Holden, James A; Brammar, Gail C; Orth, Rebecca K; Tan, Yan; Walsh, Katrina A; Fleetwood, Andrew J; Reynolds, Eric C

    2015-09-01

    Granulocyte-macrophage colony-stimulating factor (GM-CSF) and urokinase-type plasminogen activator (uPA) can contribute to the progression of chronic inflammatory diseases with possible involvement of macrophages. In this study, we investigated the role of both GM-CSF and uPA in Porphyromonas gingivalis-induced experimental periodontitis using GM-CSF-/- and uPA-/- mice. Intra-oral inoculation of wild-type (WT) C57BL/6 mice with P. gingivalis resulted in establishment of the pathogen in plaque and a significant increase in alveolar bone resorption. The infected mice also exhibited a CD11b(+) CD86(+) macrophage infiltrate into the gingival tissue, as well as P. gingivalis-specific pro-inflammatory cytokine and predominantly IgG2b antibody responses. In comparison, intra-oral inoculation of P. gingivalis did not induce bone resorption and there was significantly less P. gingivalis recovered from plaque in GM-CSF-/- and uPA-/- mice. Furthermore, P. gingivalis did not induce a macrophage gingival infiltrate or activate isolated peritoneal macrophages from the gene-deficient mice. Pro-inflammatory P. gingivalis-specific T-cell cytokine responses and serum interferon-gamma (IFN-?) and IgG2b concentrations were significantly lower in GM-CSF-/- mice. In uPA-/- mice, T-cell responses were lower but serum IFN-? and IgG2b levels were comparable with WT mice levels. These results suggest that GM-CSF and uPA are both involved in the progression of experimental periodontitis, possibly via a macrophage-dependent mechanism(s). PMID:25753270

  6. 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.

  7. Cleavage of IgG1 and IgG3 by gingipain K from Porphyromonas gingivalis may compromise host defense in progressive periodontitis

    OpenAIRE

    2011-01-01

    Degradation of immunoglobulins is an effective strategy of bacteria to evade the immune system. We have tested whether human IgG is a substrate for gingipain K of Porphyromonas gingivalis and found that the enzyme can hydrolyze subclass 1 and 3 of human IgG. The heavy chain of IgG1 was cleaved at a single site within the hinge region, generating Fab and Fc fragments. IgG3 was also cleaved within the heavy chain, but at several sites around the CH2 region. Investigation of the enzyme kinetics ...

  8. 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.

  9. The role of toll-like and protease-activated receptors in the expression of cytokines by gingival fibroblasts stimulated with the periodontal pathogen Porphyromonas gingivalis.

    Science.gov (United States)

    Palm, Eleonor; Demirel, Isak; Bengtsson, Torbjörn; Khalaf, Hazem

    2015-12-01

    Porphyromonas gingivalis is a periodontitis-associated pathogen and interactions between the bacterium and gingival fibroblasts play an important role in development and progression of periodontitis, an inflammatory disease leading to degeneration of tooth-supporting structures. Gingival fibroblasts, which expresses protease activated receptors (PARs) as well as toll-like receptors (TLRs), produces inflammatory mediators upon bacterial challenges. In this study, we elucidated the importance of PAR1, PAR2, TLR2 and TLR4 for the expression and secretion of CXCL8, interleukin-6 (IL-6), transforming growth factor-?1 (TGF-?1) and secretory leukocyte inhibitor (SLPI). Human gingival fibroblasts were transfected with small-interfering RNA against the target genes, and then stimulated with P. gingivalis wild-type W50 and W50-derived double rgp mutant E8 and kgp mutant K1A. TLR2-silencing reduced P. gingivalis-induced CXCL8 and IL-6. IL-6 was also reduced after PAR1-silencing. No effects were observed for TGF-?1. SLPI was suppressed by P. gingivalis and silencing of PAR1 as well as TLR2, gave additional suppression at the mRNA level. TLR4 was not involved in the regulation of the investigated mediators. CXCL8 and IL-6 are important for progression and development of periodontitis, leading to a chronic inflammation that may contribute to the tissue destruction that follows an exacerbated host response. Therefore, regulating the expression of TLR2 and subsequent release of CXCL8 and IL-6 in periodontitis could attenuate the tissue destruction seen in periodontitis. PMID:26318255

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

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    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.

  11. Determination of antibacterial activity of green coffee bean extract on periodontogenic bacteria like Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum and Aggregatibacter actinomycetemcomitans: An in vitro study

    Science.gov (United States)

    Bharath, Nagaraj; Sowmya, Nagur Karibasappa; Mehta, Dhoom Singh

    2015-01-01

    Background: The aim of this study was to evaluate the antibacterial activity of pure green coffee bean extract on periodonto pathogenic bacteria Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Fusobacterium nucleatum (Fn) and Aggregatibacter actinomycetemcomitans (Aa). Materials and Methods: Minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBC) were used to assess the antibacterial effect of pure green coffee bean extract against periodonto pathogenic bacteria by micro dilution method and culture method, respectively. Results: MIC values of Pg, Pi and Aa were 0.2 ?g/ml whereas Fn showed sensitive at concentration of 3.125 ?g/ml. MBC values mirrors the values same as that of MIC. Conclusion: Antimicrobial activity of pure green coffee bean extract against Pg, Pi, Fn and Aa suggests that it could be recommended as an adjunct to mechanical therapy in the management of periodontal disease. PMID:26097349

  12. Mfa4, an Accessory Protein of Mfa1 Fimbriae, Modulates Fimbrial Biogenesis, Cell Auto-Aggregation, and Biofilm Formation in Porphyromonas gingivalis

    Science.gov (United States)

    Izumigawa, Masashi; Nagano, Keiji; Yoshida, Yasuo; Kitai, Noriyuki; Lamont, Richard J.; Yoshimura, Fuminobu; Murakami, Yukitaka

    2015-01-01

    Porphyromonas gingivalis, a gram-negative obligate anaerobic bacterium, is considered to be a key pathogen in periodontal disease. The bacterium expresses Mfa1 fimbriae, which are composed of polymers of Mfa1. The minor accessory components Mfa3, Mfa4, and Mfa5 are incorporated into these fimbriae. In this study, we characterized Mfa4 using genetically modified strains. Deficiency in the mfa4 gene decreased, but did not eliminate, expression of Mfa1 fimbriae. However, Mfa3 and Mfa5 were not incorporated because of defects in posttranslational processing and leakage into the culture supernatant, respectively. Furthermore, the mfa4-deficient mutant had an increased tendency to auto-aggregate and form biofilms, reminiscent of a mutant completely lacking Mfa1. Notably, complementation of mfa4 restored expression of structurally intact and functional Mfa1 fimbriae. Taken together, these results indicate that the accessory proteins Mfa3, Mfa4, and Mfa5 are necessary for assembly of Mfa1 fimbriae and regulation of auto-aggregation and biofilm formation of P. gingivalis. In addition, we found that Mfa3 and Mfa4 are processed to maturity by the same RgpA/B protease that processes Mfa1 subunits prior to polymerization. PMID:26437277

  13. Characterization of hemin-binding protein 35 (HBP35 in Porphyromonas gingivalis: its cellular distribution, thioredoxin activity and role in heme utilization

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    Abiko Yoshimitsu

    2010-05-01

    Full Text Available Abstract Background The periodontal pathogen Porphyromonas gingivalis is an obligate anaerobe that requires heme for growth. To understand its heme acquisition mechanism, we focused on a hemin-binding protein (HBP35 protein, possessing one thioredoxin-like motif and a conserved C-terminal domain, which are proposed to be involved in redox regulation and cell surface attachment, respectively. Results We observed that the hbp35 gene was transcribed as a 1.1-kb mRNA with subsequent translation resulting in three proteins with molecular masses of 40, 29 and 27 kDa in the cytoplasm, and one modified form of the 40-kDa protein on the cell surface. A recombinant 40-kDa HBP35 exhibited thioredoxin activity in vitro and mutation of the two putative active site cysteine residues abolished this activity. Both recombinant 40- and 27-kDa proteins had the ability to bind hemin, and growth of an hbp35 deletion mutant was substantially retarded under hemin-depleted conditions compared with growth of the wild type under the same conditions. Conclusion P. gingivalis HBP35 exhibits thioredoxin and hemin-binding activities and is essential for growth in hemin-depleted conditions suggesting that the protein plays a significant role in hemin acquisition.

  14. 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.

  15. A Novel Approach for Purification and Selective Capture of Membrane Vesicles of the Periodontopathic Bacterium, Porphyromonas gingivalis: Membrane Vesicles Bind to Magnetic Beads Coated with Epoxy Groups in a Noncovalent, Species-Specific Manner

    OpenAIRE

    Nakao, Ryoma; Kikushima, Kenji; Higuchi, Hideo; Obana, Nozomu; Nomura, Nobuhiko; Bai, Dongying; Ohnishi, Makoto; Senpuku, Hidenobu

    2014-01-01

    Membrane vesicles (MVs) of Porphyromonas gingivalis are regarded as an offensive weapon of the bacterium, leading to tissue deterioration in periodontal disease. Therefore, isolation of highly purified MVs is indispensable to better understand the pathophysiological role of MVs in the progression of periodontitis. MVs are generally isolated by a conventional method based on ultracentrifugation of the bacterial culture supernatant. However, the resulting MVs are often contaminated with co-prec...

  16. 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

    Directory of Open Access Journals (Sweden)

    L Abusleme

    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.Porphyromonas gingivalis is a microorganism strongly associated with the etiology of periodontitis. This periodontal bacterium possesses an array of virulence factors, among which gingipains have a key importance, being involved with extracellular matrix destruction of periodontal tissues, modulation of host immune response and stimulation in the production of pro-inflammatory cytokines by different types of cells. These proteinases have specific affinities, being Arg-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.

  17. 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.

  18. Genome-wide transcriptome induced by Porphyromonas gingivalis LPS supports the notion of host-derived periodontal destruction and its association with systemic diseases.

    Science.gov (United States)

    Gölz, Lina; Buerfent, Benedikt C; Hofmann, Andrea; Hübner, Marc P; Rühl, Heiko; Fricker, Nadine; Schmidt, David; Johannes, Oldenburg; Jepsen, Søren; Deschner, James; Hoerauf, Achim; Nöthen, Markus M; Schumacher, Johannes; Jäger, Andreas

    2016-01-01

    Chronic periodontitis (CP) is a prevalent pathogen-associated inflammatory disorder characterized by the destruction of tooth-supporting tissues, and linked to several systemic diseases. Both the periodontopathogen Porphyromonas gingivalis (Pg), and the genetically determined host immune response, are hypothesized to play a crucial role in this association. To identify new target genes for CP and its associated systemic diseases, we investigated the transcriptome induced by Pg in human monocytes using a genome-wide approach. Monocytes were isolated from healthy male volunteers of European origin and challenged with the Pg virulence factor LPS. Array-based gene expression analysis comprising >47,000 transcripts was performed followed by pathway analyses. Transcriptional data were validated by protein and cell surface markers. LPS Pg challenge led to the significant induction of 902 transcripts. Besides known periodontitis-associated targets, several new candidates were identified (CCL23?, INDO?, GBP 1/4?, CFB?, ISG20?, MIR155HG?, DHRS9?). Moreover, various transcripts correspond to the host immune response, and have been linked to cancer, atherosclerosis and arthritis, thus highlighting the systemic impact of CP. Protein data of immunological markers validated our results. The present findings expand understanding of Pg elicited immune responses, and indicate new target genes and pathways of relevance to diagnostic and therapeutic strategies. PMID:26608307

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

    OpenAIRE

    L. Gölz; S. Memmert; Rath-Deschner, B.; A. Jäger; T. Appel; 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, ...

  20. A novel approach for purification and selective capture of membrane vesicles of the periodontopathic bacterium, Porphyromonas gingivalis: membrane vesicles bind to magnetic beads coated with epoxy groups in a noncovalent, species-specific manner.

    Science.gov (United States)

    Nakao, Ryoma; Kikushima, Kenji; Higuchi, Hideo; Obana, Nozomu; Nomura, Nobuhiko; Bai, Dongying; Ohnishi, Makoto; Senpuku, Hidenobu

    2014-01-01

    Membrane vesicles (MVs) of Porphyromonas gingivalis are regarded as an offensive weapon of the bacterium, leading to tissue deterioration in periodontal disease. Therefore, isolation of highly purified MVs is indispensable to better understand the pathophysiological role of MVs in the progression of periodontitis. MVs are generally isolated by a conventional method based on ultracentrifugation of the bacterial culture supernatant. However, the resulting MVs are often contaminated with co-precipitating bacterial appendages sheared from the live bacteria. Here, we report an intriguing property of P. gingivalis MVs--their ability to bind superparamagnetic beads coated with epoxy groups (SB-Epoxy). Analysis of fractions collected during the purification revealed that all MVs of five tested P. gingivalis stains bound to SB-Epoxy. In contrast, free fimbriae in the crude MV preparation did not bind to the SB-Epoxy. The SB-Epoxy-bound MVs were easily dissociated from the SB-Epoxy using a mild denaturation buffer. These results suggest that the surface chemistry conferred by epoxy on the beads is responsible for the binding, which is mediated by noncovalent bonds. Both the structural integrity and purity of the isolated MVs were confirmed by electron microscopy. The isolated MVs also caused cell detachment from culture dishes at a physiologically relevant concentration. Assays of competitive binding between the SB-Epoxy and mixtures of MVs from five bacterial species demonstrated that only P. gingivalis MVs could be selectively eliminated from the mixtures. We suggest that this novel approach enables efficient purification and selective elimination of P. gingivalis MVs. PMID:24830438

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

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    Kazuhisa Ouhara

    2012-03-01

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

  2. 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.

  3. 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

  4. Actinobacillus Actinomycetemcomitans y Porphyromonas Gingivales como principales patógenos periodontales

    Directory of Open Access Journals (Sweden)

    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.

  5. Bacteroides gingivalis and Bacteroides intermedius recognize different sites on human fibrinogen

    Energy Technology Data Exchange (ETDEWEB)

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

    1990-02-01

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

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

    OpenAIRE

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

    2010-01-01

    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...

  7. Porphyromonas endodontalis in chronic periodontitis: a clinical and microbiological cross-sectional study

    Directory of Open Access Journals (Sweden)

    Telma Blanca Lombardo Bedran

    2012-01-01

    Full Text Available Although previous studies have shown the presence of Porphyromonas endodontalis in chronic periodontitis associated with periapical lesions, the occurrence of this pathogen in diseased periodontal sites without periapical lesions has been poorly investigated.The aims of this study were to quantify P. endodontalis in patients with chronic periodontitis without periapical lesions, to evaluate the potential correlation of P. endodontalis with Porphyromonas gingivalis and Tannerella forsythia, and to evaluate the ability of periodontal treatment to reduce these pathogens.Patients with generalized chronic periodontitis were selected by recording clinical attachment level (CAL, probing depth (PD, and bleeding on probing (BOP. Subgingival samples from 30 diseased nonadjacent sites (CAL???5 mm, PD between 5 and 7 mm and positive BOP and 30 healthy nonadjacent sites (PD???3 mm and negative BOP were collected and subjected to microbial analysis by quantitative polymerase chain reaction (qPCR The variables of age, PD, CAL and BOP of all individuals were analyzed using the paired t-test (GrapPad Prism5®. Data of bacteria quantification were subjected to a normality test (D'Agostino-Pearson Test. For bacterial correlation analysis, the Spearman correlation was used.Our results showed that diseased sites had significantly higher levels of P. endodontalis compared to healthy sites, similar to the results obtained for P. gingivalis and T. forsythia. The numbers of all bacterial species were reduced significantly after mechanical periodontal treatment. P. endodontalis was significantly correlated with the presence of T. forsythia and P. gingivalis in the diseased group.Our results suggest that there is a high prevalence of P. endodontalis, P. gingivalis and T. forsythia in periodontitis sites and that mechanical periodontal treatment is effective at reducing the pathogens studied.

  8. 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

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

    OpenAIRE

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

    2010-01-01

    An estimated 80 million US adults have one or more types of cardiovascular diseases. Atherosclerosis is the single most important contributor to cardiovascular diseases; however, only 50% of atherosclerosis patients have currently identified risk factors. Chronic periodontitis, a common inflammatory disease, is linked to an increased cardiovascular risk. Dendritic cells (DCs) are potent antigen presenting cells that infiltrate arterial walls and may destabilize atherosclerotic plaques in card...

  10. Laser antisepsis of Phorphyromonas gingivalis in vitro with dental lasers

    Science.gov (United States)

    Harris, David M.

    2004-05-01

    It has been shown that both pulsed Nd:YAG (1064nm) and continuous diode (810nm) dental lasers kill pathogenic bacteria (laser antisepsis), but a quantitative method for determining clinical dosimetry does not exist. The purpose of this study was to develop a method to quantify the efficacy of ablation of Porphyromonas gingivalis (Pg) in vitro for two different lasers. The ablation thresholds for the two lasers were compared in the following manner. The energy density was measured as a function of distance from the output of the fiber-optic delivery system. Pg cultures were grown on blood agar plates under standard anaerobic conditions. Blood agar provides an approximation of gingival tissue for the wavelengths tested in having hemoglobin as a primary absorber. Single pulses (Nd:YAG: 100- Œs diode: 100-msec) of laser energy were delivered to Pg colonies and the energy density was increased until the appearance of a small plume was observed coincident with a laser pulse. The energy density at this point defines the ablation threshold. Ablation thresholds to a single pulse were determined for both Pg and for blood agar alone. The large difference in ablation thresholds between the pigmented pathogen and the host matrix for pulsed-Nd:YAG represented a significant therapeutic ratio and Pg was ablated without visible effect on the blood agar. Near threshold the 810-nm diode laser destroyed both the pathogen and the gel. Clinically, the pulsed Nd:YAG may selectively destroy pigmented pathogens leaving the surrounding tissue intact. The 810-nm diode laser may not demonstrate this selectivity due to its longer pulse length and greater absorption by hemoglobin.

  11. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    OpenAIRE

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

    2014-01-01

    Allium cepa L. is known to possess numerous pharmacological properties. Our aim was to examine the in vitro effects of Allium cepa L. extract (AcE) on Porphyromonas gingivalis LPS and Escherichia coli LPS-stimulated osteoclast precursor cells to determine cell viability to other future cell-based assays. Osteoclast precursor cells (RAW 264.7) were stimulated by Pg LPS (1??g/mL) and E. coli LPS (1??g/mL) in the presence or absence of different concentrations of AcE (10–1000??g/mL) for 5 days a...

  15. Porphyromonas (Bacteroides) endodontalis: its role in endodontal infections.

    Science.gov (United States)

    van Winkelhoff, A J; van Steenbergen, T J; de Graaff, J

    1992-09-01

    Porphyromonas endodontalis (formerly Bacteroides endodontalis) is a black-pigmented anaerobic Gram-negative rod which is associated with endodontal infections. It has been isolated from infected dental root canals and submucous abscesses of endodontal origin. The presence of P. endodontalis in infected dental root canals has been correlated with symptoms of an acute infection. It is occasionally found on oral mucous membranes and periodontal pockets. P. endodontalis has shown relatively low virulence in experimental monoinfections. In anaerobic mixed infections it can play an essential role. Differences in virulence between strains have been related to capsular material. On the basis of different types of capsules, three serotypes have been described. P. endodontalis is sensitive to a wide range of antibiotics, including the penicillins, the tetracyclines, and metronidazole. PMID:9796510

  16. 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; Bäckström, Malin; Johansson, Malin E V; Vester-Christensen, Malene B; Mandel, Ulla; Bennett, Eric P; Clausen, Henrik; Dahlén, Gunnar; Sroka, Aneta; Potempa, Jan; Hansson, Gunnar C

    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...

  17. Allium cepa L. and Quercetin Inhibit RANKL/Porphyromonas gingivalis LPS-Induced Osteoclastogenesis by Downregulating NF-?B Signaling Pathway.

    Science.gov (United States)

    Oliveira, Tatiane; Figueiredo, Camila A; Brito, Carlos; Stavroullakis, Alexander; Ferreira, Ana Carolina; Nogueira-Filho, Getulio; Prakki, Anuradha

    2015-01-01

    Objectives. We evaluated the in vitro modulatory effects of Allium cepa L. extract (AcE) and quercetin (Qt) on osteoclastogenesis under inflammatory conditions (LPS-induced). Methods. RAW 264.7 cells were differentiated with 30?ng/mL of RANKL, costimulated with PgLPS (1?µg/mL), and treated with AcE (50-1000?µg/mL) or Qt (1.25, 2.5, or 5?µM). Cell viability was determined by alamarBlue and protein assays. Nuclei morphology was analysed by DAPI staining. TRAP assays were performed as follows: p-nitrophenyl phosphate was used to determine the acid phosphatase activity of the osteoclasts and TRAP staining was used to evaluate the number and size of TRAP-positive multinucleated osteoclast cells. Von Kossa staining was used to measure osteoclast resorptive activity. Cytokine levels were measured on osteoclast precursor cell culture supernatants. Using western blot analysis, p-I?B? and I?B? degradation, inhibitor of NF-kappaB, were evaluated. Results. Both AcE and Qt did not affect cell viability and significantly reduced osteoclastogenesis compared to control. We observed lower production of IL-6 and IL-1? and an increased production of IL-3 and IL-4. AcE and Qt downregulated NF-?B pathway. Conclusion. AcE and Qt may be inhibitors of osteoclastogenesis under inflammatory conditions (LPS-induced) via attenuation of RANKL/PgLPS-induced NF-?B activation. PMID:26273314

  18. 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 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

  19. 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.

  20. Dietary exposure to benzoxazinoids enhances bacteria-induced monokine responses by peripheral blood mononuclear cells

    DEFF Research Database (Denmark)

    Damgaard, Dres; Jensen, Bettina Margrethe; Palarasah, Yaseelan; Nielsen, Michael Friberg Bruun; Adhikari, Khem Bahadur; Schnoor, Heidi Julius; Juel-Berg, Nanna; Poulsen, Lars K; Fomsgaard, Inge S; Nielsen, Claus Henrik

    2015-01-01

    SCOPE: To examine potentially immunomodulating effects of dietary benzoxazinoids (BXs), present in cereal grains. METHODS AND RESULTS: Nineteen healthy volunteers were randomly distributed into two groups, who received diets with high or low content of BXs for three weeks. After a week's wash-out, the groups switched diets. Peripheral blood mononuclear cells (PBMCs) were stimulated with Porphyromonas gingivalis, Escherichia coli lipopolysaccharide (LPS), or tetanus toxoid (TT). PBMCs from a heal...

  1. 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; Løvschall, Henrik; Song, Wen; Nielsen, Gitte K; Yang, Chuanxu; Wang, Qintao; Kjems, Jørgen; Gao, Shan

    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 intra...

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

    OpenAIRE

    Marquis Annie; Genovese Salvatore; Epifano Francesco; Grenier Daniel

    2012-01-01

    Abstract Background Periodontal diseases are bacterial infections leading to chronic inflammation disorders that are frequently observed in adults. In the present study, we evaluated the effect of auraptene and lacinartin, two natural oxyprenylated coumarins, on the growth, adherence properties, and collagenase activity of Porphyromonas gingivalis. We also investigated the capacity of these compounds to reduce cytokine and matrix metalloproteinase (MMP) secretion by lipopolysaccharide (LPS)-s...

  3. 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...

  4. A Murine Model of Lipopolysaccharide-Induced Peri-Implant Mucositis and Peri-Implantitis.

    Science.gov (United States)

    Pirih, Flavia Q; Hiyari, Sarah; Leung, Ho-Yin; Barroso, Ana D V; Jorge, Adrian C A; Perussolo, Jeniffer; Atti, Elisa; Lin, Yi-Ling; Tetradis, Sotirios; Camargo, Paulo M

    2015-10-01

    Dental implants are a widely used treatment option for tooth replacement. However, they are susceptible to inflammatory diseases such as peri-implant mucositis and peri-implantitis, which are highly prevalent and may lead to implant loss. Unfortunately, the understanding of the pathogenesis of peri-implant mucositis and peri-implantitis is fragmented and incomplete. Therefore, the availability of a reproducible animal model to study these inflammatory diseases would facilitate the dissection of their pathogenic mechanisms. The objective of this study is to propose a murine model of experimental peri-implant mucositis and peri-implantitis. Screw-shaped titanium implants were placed in the upper healed edentulous alveolar ridges of C57BL/6J mice 8 weeks after tooth extraction. Following 4 weeks of osseointegration, Porphyromonas gingivalis -lipolysaccharide (LPS) injections were delivered to the peri-implant soft tissues for 6 weeks. No-injections and vehicle injections were utilized as controls. Peri-implant mucositis and peri-implantitis were assessed clinically, radiographically (microcomputerized tomograph [CT]), and histologically following LPS-treatment. LPS-injections resulted in a significant increase in soft tissue edema around the head of the implants as compared to the control groups. Micro-CT analysis revealed significantly greater bone loss in the LPS-treated implants. Histological analysis of the specimens demonstrated that the LPS-group had increased soft tissue vascularity, which harbored a dense mixed inflammatory cell infiltrate, and the bone exhibited noticeable osteoclast activity. The induction of peri-implant mucositis and peri-implantitis in mice via localized delivery of bacterial LPS has been demonstrated. We anticipate that this model will contribute to the development of more effective preventive and therapeutic approaches for these 2 conditions. PMID:24967609

  5. Effect of hemin on the physiology and virulence of Bacteroides gingivalis W50.

    OpenAIRE

    McKee, A.S.; McDermid, A S; Baskerville, A.; Dowsett, A. B.; Ellwood, D C; Marsh, P D

    1986-01-01

    Bacteroides gingivalis W50 was grown in a chemostat under steady-state conditions at pH 7.5 +/- 0.2 and a constant growth rate of 6.9 h for periods of up to 6 weeks (146 bacterial generations) in a complex medium. Hemin was capable of limiting the growth of cells up to a concentration of approximately 0.5 micrograms/ml since higher concentrations of hemin did not increase cell yields; cells grew in the absence of exogenously added vitamin K1. Only a limited number of amino acids was metaboliz...

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

    Directory of Open Access Journals (Sweden)

    Marquis Annie

    2012-06-01

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

  7. Immunochemical characterization of Brucella lipopolysaccharides and polysaccharides.

    OpenAIRE

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

    1981-01-01

    Purified lipopolysaccharide (LPS) extracted with phenol-water from smooth Brucella abortus was hydrolyzed with 1% acetic acid at 100 degrees C. The degraded polysaccharide (AH) released gave reactions of identity with the native polysaccharide hapten (NH) in phenol-water- or trichloroacetic acid-extracted endotoxin preparations of B. abortus and with the polysaccharide (poly B) extracted by trichloroacetic acid from rough B. melitensis strain B115. Poly B was present in the soluble cytoplasmi...

  8. Immunochemical identification of Brucella abortus lipopolysaccharide epitopes.

    OpenAIRE

    Rojas, N.; Freer, E; Weintraub, A; Ramirez, M.; Lind, S.; Moreno, E.

    1994-01-01

    Sera from Brucella abortus-infected and -vaccinated bovines recognized four lipopolysaccharide (LPS) determinants: two in the O-polysaccharide (A and C), one in the core oligosaccharide from rough Brucella LPS (R), and one in lipid A (LA). From 46 different hybridomas secreting monoclonal antibodies (MAbs) against various LPS moieties, 9 different specificities were identified. Two epitopes, A and C/Y, were present in the O-polysaccharide. Two epitopes were found in the core oligosaccharide (...

  9. Acetate supplementation attenuates lipopolysaccharide-induced neuroinflammation

    OpenAIRE

    Reisenauer, Chris J.; Bhatt, Dhaval P; Mitteness, Dane J.; Slanczka, Evan R.; Gienger, Heidi M.; Watt, John A.; Rosenberger, Thad A.

    2011-01-01

    Glyceryl triacetate (GTA), a compound effective at increasing circulating and tissue levels of acetate was used to treat rats subjected to a continual 28 day intra-ventricular infusion of bacterial lipopolysaccharide (LPS). This model produces a neuroinflammatory injury characterized by global neuroglial activation and a decrease in choline acetyltransferase immunoreactivity in the basal forebrain. During the LPS infusion, rats were given a daily treatment of either water or GTA at a dose of ...

  10. Phospholipids and lipopolysaccharide of Aeromonas hydrophila.

    OpenAIRE

    Howard, S. P.; Buckley, J T

    1985-01-01

    The phospholipids and lipopolysaccharide of Aeromonas hydrophila were characterized. Phosphatidylethanolamine and phosphatidylglycerol were the major phospholipid components. The outer membrane contained more phosphatidylethanolamine and less phosphatidylglycerol than the inner membrane, and the phospholipids of the outer membrane contained a higher proportion of saturated fatty acids. Only four fatty acids (C14:0, C16:0, C16:1, and C18:1) were found in the phospholipids. The lipopolysacchari...

  11. Blastogenic response of bovine lymphocytes to Brucella abortus lipopolysaccharide.

    OpenAIRE

    Baldwin, C. L.; Winter, A J

    1985-01-01

    Brucella abortus lipopolysaccharide was tested in a blastogenesis assay with unfractionated and nylon wool-separated peripheral blood lymphocytes of Brucella-naive cattle and cattle immunized with B. abortus. Our results indicated that in cattle the lipopolysaccharide of B. abortus is not a B-cell mitogen. In immunized animals it stimulated predominantly nylon wool-adherent cells. The lipopolysaccharide of Escherichia coli O128:B12, in contrast, induced a substantially greater proliferative r...

  12. 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.

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

    Directory of Open Access Journals (Sweden)

    Silvio Favoreto Junior

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

  14. Biological activities of Brucella abortus lipopolysaccharides.

    OpenAIRE

    Moreno, E.; Berman, D. T.; Boettcher, L A

    1981-01-01

    Purified lipopolysaccharide (LPS) from smooth (s) and rough (R) strains of Brucella abortus and lipid A isolated from S-LPS by mild acid hydrolysis were examined in several assays of biological activity. Brucella S- and R-LPSs and Brucella lipid A activated the complement cascade. Previously reported mitogenic activation by Brucella LPSs of spleen cells from endotoxin-resistant C3H/HeJ mice was confirmed and also produced by isolated Brucella lipid A. Mitogenicity was not inhibited by polymyx...

  15. [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

  16. 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.

  17. Immunogenic mimics of Brucella lipopolysaccharide epitopes.

    Science.gov (United States)

    Beninati, Concetta; Garibaldi, Manuela; Lo Passo, Carla; Mancuso, Giuseppe; Papasergi, Salvatore; Garufi, Gabriella; Pernice, Ida; Teti, Giuseppe; Felici, Franco

    2009-10-01

    Brucella melitensis and Brucella abortus are responsible for brucellosis in bovine and ovine species and for Malta fever in humans. The lipopolysaccharide (LPS) of Brucella is an important virulence factor and can elicit protective antibodies. Because of their potential importance in vaccine design and in serological diagnosis, we developed peptides mimicking the antigenic properties of distinctive antigenic determinants of Brucella LPS. These peptides were selected from several phage display random peptide libraries for their ability to bind monoclonal antibodies directed against the A- or C-type epitopes of Brucella LPS. Plasmids encoding for two of the isolated peptides induced, after DNA immunization, LPS-specific antibody responses. Although these responses were only moderate in extent, these data further suggest the feasibility of using peptide mimics of carbohydrate epitopes as immunogens, a property which may be useful in the design of novel anti-Brucella vaccines. PMID:19631246

  18. Structure and Effects of Cyanobacterial Lipopolysaccharides

    Directory of Open Access Journals (Sweden)

    Prasannavenkatesh Durai

    2015-07-01

    Full Text Available Lipopolysaccharide (LPS is a component of the outer membrane of mainly Gram-negative bacteria and cyanobacteria. The LPS molecules from marine and terrestrial bacteria show structural variations, even among strains within the same species living in the same environment. Cyanobacterial LPS has a unique structure, since it lacks heptose and 3-deoxy-d-manno-octulosonic acid (also known as keto-deoxyoctulosonate (KDO, which are present in the core region of common Gram-negative LPS. In addition, the cyanobacterial lipid A region lacks phosphates and contains odd-chain hydroxylated fatty acids. While the role of Gram-negative lipid A in the regulation of the innate immune response through Toll-like Receptor (TLR 4 signaling is well characterized, the role of the structurally different cyanobacterial lipid A in TLR4 signaling is not well understood. The uncontrolled inflammatory response of TLR4 leads to autoimmune diseases such as sepsis, and thus the less virulent marine cyanobacterial LPS molecules can be effective to inhibit TLR4 signaling. This review highlights the structural comparison of LPS molecules from marine cyanobacteria and Gram-negative bacteria. We discuss the potential use of marine cyanobacterial LPS as a TLR4 antagonist, and the effects of cyanobacterial LPS on humans and marine organisms.

  19. Structure and Effects of Cyanobacterial Lipopolysaccharides.

    Science.gov (United States)

    Durai, Prasannavenkatesh; Batool, Maria; Choi, Sangdun

    2015-07-01

    Lipopolysaccharide (LPS) is a component of the outer membrane of mainly Gram-negative bacteria and cyanobacteria. The LPS molecules from marine and terrestrial bacteria show structural variations, even among strains within the same species living in the same environment. Cyanobacterial LPS has a unique structure, since it lacks heptose and 3-deoxy-d-manno-octulosonic acid (also known as keto-deoxyoctulosonate (KDO)), which are present in the core region of common Gram-negative LPS. In addition, the cyanobacterial lipid A region lacks phosphates and contains odd-chain hydroxylated fatty acids. While the role of Gram-negative lipid A in the regulation of the innate immune response through Toll-like Receptor (TLR) 4 signaling is well characterized, the role of the structurally different cyanobacterial lipid A in TLR4 signaling is not well understood. The uncontrolled inflammatory response of TLR4 leads to autoimmune diseases such as sepsis, and thus the less virulent marine cyanobacterial LPS molecules can be effective to inhibit TLR4 signaling. This review highlights the structural comparison of LPS molecules from marine cyanobacteria and Gram-negative bacteria. We discuss the potential use of marine cyanobacterial LPS as a TLR4 antagonist, and the effects of cyanobacterial LPS on humans and marine organisms. PMID:26198237

  20. 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.

  1. 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.

  2. 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

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

    OpenAIRE

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

    1996-01-01

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

  4. Innate cell recognition of Lipopolysaccharide and other bacterial molecules

    OpenAIRE

    Poussin, Carine

    2004-01-01

    Une étape critique de la pathogenèse du sepsis est la reconnaissance des molécules bactériennes telles que les lipopolysaccharides (LPS) par les cellules de l'immunité innée. L'interaction du LPS, complexé à la "lipopolysaccharide binding protein" (LBP), avec les récepteurs CD14 et TLR4/MD-2 présents à la surface des cellules myéloïdes induit l'activation cellulaire. Parallèlement, afin d'être détoxifié, le LPS est internalisé de façon dépendante du CD14 par une voie de macropinocytose dans l...

  5. Innate cell recognition of Lipopolysaccharide and other bacterial molecules

    OpenAIRE

    Poussin, Carine; Chevrolet, Jean-Claude; Pugin, Jérôme

    2005-01-01

    Une étape critique de la pathogenèse du sepsis est la reconnaissance des molécules bactériennes telles que les lipopolysaccharides (LPS) par les cellules de l'immunité innée. L'interaction du LPS, complexé à la "lipopolysaccharide binding protein" (LBP), avec les récepteurs CD14 et TLR4/MD-2 présents à la surface des cellules myéloïdes induit l'activation cellulaire. Parallèlement, afin d'être détoxifié, le LPS est internalisé de façon dépendante du CD14 par une voie de macropinocytose dans l...

  6. LipidBank - Lipopolysaccharide(CLS5428) [LipidBank

    Lifescience Database Archive (English)

    Full Text Available Lipopolysaccharide DATA No : CLS5428 INFORMANT : Shoichi Kusumoto NAME : COMMON NAME: the lipopo ... cillus pleuropneumoniae serotypes 1, 2, 5a and the genome ... strain 5b SYMBOL: FORMULA: C57H102C48 MOL.WT (aver ... acillus pleuropneumonia serotypes 1, 2, 5a and the genome ... strain 5b CHEMICAL SYNTHESIS METABOLISM GENETIC IN ...

  7. LipidBank - Lipopolysaccharide(CLS5427) [LipidBank

    Lifescience Database Archive (English)

    Full Text Available Lipopolysaccharide DATA No : CLS5427 INFORMANT : Shoichi Kusumoto NAME : COMMON NAME: the lipopo ... cillus pleuropneumoniae serotypes 1, 2, 5a and the genome ... strain 5b SYMBOL: FORMULA: C69H120NO56 MOL.WT (ave ... acillus pleuropneumonia serotypes 1, 2, 5a and the genome ... strain 5b CHEMICAL SYNTHESIS METABOLISM GENETIC IN ...

  8. LipidBank - Lipopolysaccharide(CLS5426) [LipidBank

    Lifescience Database Archive (English)

    Full Text Available Lipopolysaccharide DATA No : CLS5426 INFORMANT : Shoichi Kusumoto NAME : COMMON NAME: the lipopo ... cillus pleuropneumoniae serotypes 1, 2, 5a and the genome ... strain 5b SYMBOL: FORMULA: C63H112O53 MOL.WT (aver ... acillus pleuropneumonia serotypes 1, 2, 5a and the genome ... strain 5b CHEMICAL SYNTHESIS METABOLISM GENETIC IN ...

  9. LipidBank - Lipopolysaccharide(CLS1731) [LipidBank

    Lifescience Database Archive (English)

    Full Text Available Lipopolysaccharide DATA No : CLS1731 INFORMANT : Yoshio Kumazawa NAME : 2'N-3''(tetradecenoyloxy ... spectra of R. sphaeroides DPLA[Spectrum 0070]High-energy ... CID spectra of carboxylate anions at m/z 187[Spect ... spectrum of R. sphaeroides DPLA[Spectrum 0072]High-energy ... CID spectrum of [M+Na]+[Spectrum 0073]Low-energy ... C ...

  10. 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

  11. Growth characteristics and a novel method for identification (the WEE-TAB system) of Porphyromonas species isolated from infected dog and cat bite wounds in humans.

    OpenAIRE

    Hudspeth, M K; Hunt Gerardo, S; Citron, D. M.; Goldstein, E J

    1997-01-01

    Thirty-nine clinical isolates of Porphyromonas species recovered from infected cat and dog bite wounds in humans and eight American Type Culture Collection and National Collection of Type Cultures type strains were characterized by using the API ZYM system, the RapID ANA II system, and conventional biochemical methods. Growth characteristics on various agar media were compared. All strains grew on brucella blood agar supplemented with vitamin K1 and hemin and on brucella laked blood agar supp...

  12. 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.

  13. Physical and morphological characteristics of eucaryotic ribosomes and lipopolysaccharide complexes.

    OpenAIRE

    Phillips, M.; Brogden, K.A.

    1987-01-01

    Lipopolysaccharides (LPS) from Pasteurella multocida or Brucella abortus were complexed with Aspergillus fumigatus ribosomes by mixing and fixation for 3 days in 3.8% formaldehyde. To investigate the nature of their physical association, ribosomes, LPS, and ribosome-LPS complexes were (i) centrifuged in CsCl gradients to determine buoyant densities, (ii) examined by electron microscopy, and (iii) monitored by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Ribosomes were found to b...

  14. Role of lipopolysaccharide in virulence of Pseudomonas aeruginosa.

    OpenAIRE

    Cryz, S J; Pitt, T. L.; Fürer, E.; Germanier, R.

    1984-01-01

    The role of lipopolysaccharide (LPS) in the virulence of Pseudomonas aeruginosa was studied. The virulence of several P. aeruginosa strains for burned mice was found to be directly related to the dispersion of LPS into either the phenol or the water phase after extraction. Virulence decreased as the proportion of LPS recovered from the phenol phase increased. No similar correlation was observed when several other strain characteristics were investigated. This phenomenon was studied in greater...

  15. 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

  16. Binding and neutralization of bacterial lipopolysaccharide by colistin nonapeptide.

    OpenAIRE

    Warren, H S; Kania, S A; Siber, G R

    1985-01-01

    Polymyxin nonapeptides, proteolytic derivatives of polymyxin antibiotics, are less toxic than their parent compounds but retain some of their antibacterial activities. To confirm and expand observations that polymyxin nonapeptides have anti-endotoxin activity, we studied the ability of colistin nonapeptide to bind to bacterial lipopolysaccharide (LPS) and to inhibit the effects of LPS on Limulus amoebocyte lysate and lymphocyte mitogenicity. Colistin nonapeptide was purified by high-pressure ...

  17. Lipopolysaccharide-induced leptin release is neurally controlled

    OpenAIRE

    Mastronardi, C. A.; W.H. Yu; V. K. SRIVASTAVA; Dees, W. L.; McCann, S M

    2001-01-01

    Our hypothesis is that leptin release is controlled neurohormonally. Conscious, male rats bearing indwelling, external, jugular catheters were injected with the test drug or 0.9% NaCl (saline), and blood samples were drawn thereafter to measure plasma leptin. Anesthesia decreased plasma leptin concentrations within 10 min to a minimum at 120 min, followed by a rebound at 360 min. Administration (i.v.) of lipopolysaccharide (LPS) increased plasma leptin to almost tw...

  18. Inhibitory Effects of Ketamine on Lipopolysaccharide-Induced Microglial Activation

    OpenAIRE

    Joen-Rong Sheu; Duen-Suey Chou; George Hsiao; Jie-Jen Lee; Cheng-Ying Hsieh; Yi Chang

    2009-01-01

    Microglia activated in response to brain injury release neurotoxic factors including nitric oxide (NO) and proinflammatory cytokines such as tumor necrosis factor-? (TNF-?) and interleukin-1? (IL-1?). Ketamine, an anesthetic induction agent, is generally reserved for use in patients with severe hypotension or respiratory depression. In this study, we found that ketamine (100 and 250 ?M) concentration-dependently inhibited lipopolysaccharide (LPS)-induced NO and IL-1? release in primary cu...

  19. 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 cardiomega...

  20. Sickness behaviour after lipopolysaccharide treatment in ghrelin deficient mice

    OpenAIRE

    Szentirmai, Éva; Krueger, James M.

    2013-01-01

    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, ...

  1. Pulmonary collectins selectively permeabilize model bacterial membranes containing rough lipopolysaccharide

    OpenAIRE

    Kuzmenko, Alexander I.; Wu, Huixing; McCormack, Francis X

    2006-01-01

    We have reported that Gram negative organisms decorated with rough lipopolysaccharide (LPS) are particularly susceptible to the direct antimicrobial actions of the pulmonary collectins, surfactant proteins A (SP-A) and D (SP-D). In this study, we examined the lipid and LPS components required for the permeabilizing effects of the collectins on model bacterial membranes. Liposomes composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE), with or without rough E. coli LPS (J5), ...

  2. Recognition of lipopolysaccharide pattern by TLR4 complexes

    OpenAIRE

    Park, Beom Seok; Lee, Jie-Oh

    2013-01-01

    Lipopolysaccharide (LPS) is a major component of the outer membrane of Gram-negative bacteria. Minute amounts of LPS released from infecting pathogens can initiate potent innate immune responses that prime the immune system against further infection. However, when the LPS response is not properly controlled it can lead to fatal septic shock syndrome. The common structural pattern of LPS in diverse bacterial species is recognized by a cascade of LPS receptors and accessory proteins, LPS bindin...

  3. 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 histopathological d...

  4. 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 (P<0.05) than in pigs fed the other diets. Whereas pigs fed diet WB had about 2 to 22 times higher abundance (P<0.05) of Megasphaera elsdenii in feces and about six times higher abundance (P<0.05) of Lactobacillus 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

  5. SIRT2 ameliorates lipopolysaccharide-induced inflammation in macrophages

    International Nuclear Information System (INIS)

    Highlights: • Knockout of SIRT2 attenuates lipopolysaccharide-induced iNOS expression. • Lipopolysaccharide-induced NO production is decreased in SIRT2 KO macrophage. • SIRT2 deficiency suppresses lipopolysaccharide-induced ROS production in macrophage. • M1-macrophage related factors are decreased in SIRT2 deficient cells. • SIRT2 deficiency decreases lipopolysaccharide-induced activation of NF?B. - Abstract: Introduction: SIRT2 is a NAD(+)-dependent deacetylases and associated with numerous processes such as infection, carcinogenesis, DNA damage and cell cycle regulation. However, the role of SIRT2 in inflammatory process in macrophage remains unclear. Materials and methods: In the present study, we have evaluated the regulatory effects of SIRT2 in lipopolysaccharide (LPS)-stimulated macrophages isolated from SIRT2 knockout (KO) and wild type (WT) mice or Raw264.7 macrophage cells. As inflammatory parameters, expression of inducible nitric oxide synthase (iNOS), the productions of nitric oxide, reactive oxygen species (ROS) and M1-macrophage-related factors were evaluated. We also examined the effects of SIRT2 on activation of nuclear factor-kappaB (NF?B) signaling. Results: SIRT2 deficiency inhibits LPS-induced iNOS mRNA and protein expression in bone marrow derived macrophages. SIRT2-siRNA transfection also suppressed LPS-induced iNOS expression in Raw264.7 macrophage cells. Bone marrow derived macrophages isolated from SIRT2 KO mice produced lower nitric oxide and expressed lower levels of M1-macrophage related markers including iNOS and CD86 in response to LPS than WT mice. Decrease of SIRT2 reduced the LPS-induced reactive oxygen species production. Deficiency of SIRT2 resulted in inhibition of NF?B activation through reducing the phosphorylation and degradation of I?B?. The phosphorylation and nuclear translocation of p65 was significantly decreased in SIRT2-deficient macrophages after LPS stimulation. Discussion: Our data suggested that deficiency of SIRT2 ameliorates iNOS, NO expression and reactive oxygen species production with suppressing LPS-induced activation of NF?B in macrophages

  6. Short communication: Distribution of Porphyromonas gulae fimA genotypes in oral specimens from dogs with mitral regurgitation.

    Science.gov (United States)

    Shirai, Mitsuyuki; Nomura, Ryota; Kato, Yukio; Murakami, Masaru; Kondo, Chihiro; Takahashi, Soraaki; Yamasaki, Yoshie; Matsumoto-Nakano, Michiyo; Arai, Nobuaki; Yasuda, Hidemi; Nakano, Kazuhiko; Asai, Fumitoshi

    2015-10-01

    Porphyromonas gulae, a suspected pathogen for periodontal disease in dogs, possesses approximately 41-kDa fimbriae (FimA) that are encoded by the fimA gene. In the present study, the association of fimA genotypes with mitral regurgitation (MR) was investigated. Twenty-five dogs diagnosed with MR (age range 6-13 years old, average 10.8years) and 32 healthy dogs (8-15 years old, average 10.8years) were selected at the participating clinics in a consecutive manner during the same time period. Oral swab specimens were collected from the dogs and bacterial DNA was extracted, then polymerase chain reaction analysis was performed using primers specific for each fimA genotype, with the dominant genotype determined. The rate for genotype C dominant specimens was 48.0% in the MR group, which was significantly higher than that in the control group (18.8%) (P <0.05). These results suggest that P. gulae fimA genotype C is associated with MR. PMID:26412519

  7. Dietary exposure to benzoxazinoids enhances bacteria-induced monokine responses by peripheral blood mononuclear cells

    DEFF Research Database (Denmark)

    Damgaard, Dres; Jensen, Bettina Margrethe

    2015-01-01

    SCOPE: To examine potentially immunomodulating effects of dietary benzoxazinoids (BXs), present in cereal grains. METHODS AND RESULTS: Nineteen healthy volunteers were randomly distributed into two groups, who received diets with high or low content of BXs for three weeks. After a week's wash-out, the groups switched diets. Peripheral blood mononuclear cells (PBMCs) were stimulated with Porphyromonas gingivalis, Escherichia coli lipopolysaccharide (LPS), or tetanus toxoid (TT). PBMCs from a healthy donor received the same stimuli in presence of serum from each participant receiving BXs. The production of monokines, T-cell cytokines and T-helper cell proliferation were assessed. A three-week diet with high BX content enhanced IL-1? responses against LPS and P. gingivalis, as well as TNF-? response against P. gingivalis, after 24 hours of stimulation. Moreover, IL-6 was found to be increased after 7 days of stimulation with LPS. No effect was observed on T-cell cytokines or proliferation. BX levels in serum after a single meal did not modify cytokine responses. CONCLUSIONS: High dietary intake of BXs enhances bacteria-induced production of pro-inflammatory monokines by PBMCs, but not T-cell responses; presumably due to intrinsic changes within PBMCs, built up over three weeks of BX-rich diet, rather than to immediate effects of BXs contained in serum. This article is protected by copyright. All rights reserved.

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

    OpenAIRE

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

    2010-01-01

    Lipopolysaccharide exerts many effects on many cell lines, including cytokine secretion, and cell apoptosis and necrosis. We investigated the in vitro effects of lipopolysaccharide on apoptosis of cultured human dental pulp cells and the expression of Bcl-2 and Bax. Dental pulp cells showed morphologies typical of apoptosis after exposure to lipopolysaccharide. Flow cytometry showed that the rate of apoptosis of human dental pulp cells increased with increasing lipopolysaccharide concentratio...

  9. Btk Regulates Macrophage Polarization in Response to Lipopolysaccharide

    OpenAIRE

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

    2014-01-01

    Bacterial Lipopolysaccharide (LPS) is a strong inducer of inflammation and does so by inducing polarization of macrophages to the classic inflammatory M1 population. Given the role of Btk as a critical signal transducer downstream of TLR4, we investigated its role in M1/M2 induction. In Btk deficient (Btk2\\2) mice we observed markedly reduced recruitment of M1 macrophages following intraperitoneal administration of LPS. Ex vivo analysis demonstrated an impaired ability of Btk2/2 macrophages t...

  10. 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.

  11. Lipopolysaccharide-induced acute renal failure in conscious rats

    DEFF Research Database (Denmark)

    Jonassen, Thomas E N; Graebe, Martin; Promeneur, Dominique; Nielsen, Søren; Christensen, Sten; Olsen, Niels Vidiendal

    2002-01-01

    In conscious, chronically instrumented rats we examined 1) renal tubular functional changes involved in lipopolysaccharide (LPS)-induced acute renal failure; 2) the effects of LPS on the expression of selected renal tubular water and sodium transporters; and 3) effects of milrinone, a phosphodiesterase type 3 (PDE3) inhibitor, and Ro-20-1724, a PDE4 inhibitor, on LPS-induced changes in renal function. Intravenous infusion of LPS (4 mg/kg b.wt. over 1 h) caused an immediate decrease in glomerular...

  12. Antioxidant Effects of Cranberry Powder in Lipopolysaccharide Treated Hypercholesterolemic Rats

    OpenAIRE

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

    2014-01-01

    This study was conducted to investigate the effects of cranberry power on antioxidant defense system in rats fed an atherogenic diet and injected with lipopolysaccharide (LPS). Sprague-Dawley rats were divided into the following 5 groups: normal diet+saline (NS), atherogenic diet+saline (AS), atherogenic diet+LPS (AL), atherogenic diet with 5% cranberry powder+LPS (AL-C5), and atherogenic diet with 10% cranberry powder+LPS (AL-C10). Total antioxidant status measured by ferric reducing ability...

  13. A Second Outer-Core Region in Klebsiella pneumoniae Lipopolysaccharide

    OpenAIRE

    Regué, Miguel; Izquierdo, Luis; Fresno, Sandra; Piqué, Núria; Corsaro, Maria Michela; Naldi, Teresa; De Castro, Cristina; Waidelich, Dietmar; Merino, Susana; Tomás, Juan M.

    2005-01-01

    Up to now only one major type of core oligosaccharide has been found in the lipopolysaccharide of all Klebsiella pneumoniae strains analyzed. Applying a different screening approach, we identified a novel Klebsiella pneumoniae core (type 2). Both Klebsiella core types share the same inner core and the outer-core-proximal disaccharide, GlcN-(1,4)-GalA, but they differ in the GlcN substituents. In core type 2, the GlcpN residue is substituted at the O-4 position by the disaccharide ?-Glcp(1-6)-...

  14. 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 ...

  15. Lipopolysaccharide stabilizes the crystal packing of the ABC transporter MsbA

    International Nuclear Information System (INIS)

    The use of lipopolysaccharide in the crystallization of the integral membrane protein MsbA was critical to the overall stability of the crystals and led to an increase in diffraction resolution. The ABC transporter MsbA is an integral membrane protein involved in the transport of lipid A and lipopolysaccharides to the outer leaflet of the inner membrane in bacteria. Here, the critical role of the natural substrate lipopolysaccharide in the crystallization and diffraction quality of MsbA crystals is reported. Initial crystals grown in complex with ATP–vanadate alone diffracted to ?9 Å. Screening of the natural substrate lipopolysaccharides led to the crystallization of MsbA in complex with ADP–vanadate and Ra lipopolysaccharide. The increased order within the crystal lattice allowed structure determination to 4.2 Å

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

    Science.gov (United States)

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

    2015-06-01

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

  17. Deciphering the dual effect of lipopolysaccharides from plant pathogenic Pectobacterium.

    Science.gov (United States)

    Mohamed, Kettani-Halabi; Daniel, Tran; Aurélien, Dauphin; El-Maarouf-Bouteau, Hayat; Rafik, Errakhi; Arbelet-Bonnin, Delphine; Biligui, Bernadette; Florence, Val; Mustapha, Ennaji Moulay; François, Bouteau

    2015-01-01

    Lipopolysaccharides (LPS) are a component of the outer cell surface of almost all Gram-negative bacteria and play an essential role for bacterial growth and survival. Lipopolysaccharides represent typical microbe-associated molecular pattern (MAMP) molecules and have been reported to induce defense-related responses, including the expression of defense genes and the suppression of the hypersensitive response in plants. However, depending on their origin and the challenged plant, LPS were shown to have complex and different roles. In this study we showed that LPS from plant pathogens Pectobacterium atrosepticum and Pectobacterium carotovorum subsp. carotovorum induce common and different responses in A. thaliana cells when compared to those induced by LPS from non-phytopathogens Escherichia coli and Pseudomonas aeruginosa. Among common responses to both types of LPS are the transcription of defense genes and their ability to limit of cell death induced by Pectobacterium carotovorum subsp carotovorum. However, the differential kinetics and amplitude in reactive oxygen species (ROS) generation seemed to regulate defense gene transcription and be determinant to induce programmed cell death in response to LPS from the plant pathogenic Pectobacterium. These data suggest that different signaling pathways could be activated by LPS in A. thaliana cells. PMID:25760034

  18. Interactions of Bacterial Lipopolysaccharides with Gold Nanorod Surfaces Investigated by Refractometric Sensing.

    Science.gov (United States)

    Abadeer, Nardine S; Fülöp, Gerg?; Chen, Si; Käll, Mikael; Murphy, Catherine J

    2015-11-11

    The interface between nanoparticles and bacterial surfaces is of great interest for applications in nanomedicine and food safety. Here, we demonstrate that interactions between gold nanorods and bacterial surface molecules are governed by the nanoparticle surface coating. Polymer-coated gold nanorod substrates are exposed to lipopolysaccharides extracted from Pseudomonas aeruginosa, Salmonella enterica and Escherichia coli, and attachment is monitored using localized surface plasmon resonance refractometric sensing. The number of lipopolysaccharide molecules attached per nanorod is calculated from the shift in the plasmon maximum, which results from the change in refractive index after analyte binding. Colloidal gold nanorods in water are also incubated with lipopolysaccharides to demonstrate the effect of lipopolysaccharide concentration on plasmon shift, ?-potential, and association constant. Both gold nanorod surface charge and surface chemistry affect gold nanorod-lipopolysaccharide interactions. In general, anionic lipopolysaccharides was found to attach more effectively to cationic gold nanorods than to neutral or anionic gold nanorods. Some variation in lipopolysaccharide attachment is also observed between the three strains studied, demonstrating the potential complexity of bacteria-nanoparticle interactions. PMID:26488238

  19. 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.

  20. 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

  1. 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

  2. Propolis antimicrobial activity against periodontopathic bacteria

    OpenAIRE

    Gebara Elaine C.E.; Lima Luiz A.; Mayer Marcia P.A.

    2002-01-01

    Propolis extract antimicrobial activity against periodontopathic (ATCC) bacteria was investigated "in vitro". Bacterial strains tested were: Prevotella intermedia, Prevotella melaninogenica, Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Capnocytophaga gingivalis and Fusobacterium nucleatum. Minimal inhibitory concentration (MIC) for the strains tested was determined using the method of broth dilution with the propolis extract in serial concentrations. Results showed MIC of 1...

  3. 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???????? ...

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

  5. 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

  6. Effects of D-003 on Lipopolysaccharides-induced Osteonecrosis in Rabbits

    OpenAIRE

    Noa, Miriam; Valle, M; Mendoza, Sarahí; Mas, Rosa; Mendoza, Nilda

    2011-01-01

    D-003, a mixture of high molecular weight acids, inhibits cholesterol synthesis prior to mevalonate and prevents osteoporosis induced by ovariectomy in rats, and both osteoporosis and osteonecrosis induced by corticoids in rats. The aim of this study was to investigate effects of D-003 on lipopolysaccharides-induced osteonecrosis in rabbits. Animals were randomized into 5 groups: a sham and four groups injected with lipopolysaccharides: one treated orally with vehicle and three with D-003 (5,...

  7. Unilamellar liposomes modulate secretion of tumor necrosis factor by lipopolysaccharide-stimulated macrophages.

    OpenAIRE

    Brisseau, G F; Kresta, A; Schouten, D.(TRIUMF, Vancouver, BC, Canada; Department of Physics and Astronomy, York University, Toronto, ON, Canada); Bohnen, J M; Shek, P.N.; Fok, E; Rotstein, O D

    1994-01-01

    Liposomal encapsulation of antimicrobial agents has been used to improve drug delivery, particularly against intracellular pathogens. The effect of unilamellar liposomes on macrophage activation in response to Escherichia coli lipopolysaccharide was examined. Liposomes caused a dose- and time-dependent inhibition of tumor necrosis factor release by lipopolysaccharide-treated cells. The accumulation of tumor necrosis factor mRNA transcripts was unaffected, suggesting a posttranscriptional mech...

  8. Baclofen influences lipopolysaccharide-mediated interleukin-6 release from murine pituicytes

    DEFF Research Database (Denmark)

    Kjeldsen, Tine H; Hansen, Erik W; Christensen, Jens D; Moesby, Lise

    2002-01-01

    Pituicytes, the glial cells of the neurohypophysis, secrete interleukin-6 upon stimulation with various inflammatory mediators, i.e. lipopolysaccharide. Previous studies have identified several receptors on pituicytes. This study investigates the effect of GABA(B) receptor activation on interleukin-6 release from pituicytes. Cultured murine pituicytes were stimulated for 24 h with lipopolysaccharide (0.5 ng/ml) to give a significant interleukin-6 release compared to control. The interleukin-6 re...

  9. Pulmonary toxicity of endotoxins: comparison of lipopolysaccharides from various bacterial species.

    OpenAIRE

    Helander, I; Saxén, H; Salkinoja-Salonen, M; Rylander, R.

    1982-01-01

    Lipopolysaccharides from three gram-negative bacteria isolated from bale cotton and piggery air were analyzed for their chemical composition, and their pulmonary toxicity for guinea pigs, lethal toxicity for mice, and pyrogenicity for rabbits were measured. Lipopolysaccharides from Enterobacter agglomerans and Citrobacter freundii had closely related chemical compositions; both were pyrogenic for rabbits and caused a dose-dependent influx of polymorphonuclear leukocytes into the airways of gu...

  10. 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.

  11. 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

  12. Activity of Host Antimicrobials against Multidrug-Resistant Acinetobacter baumannii Acquiring Colistin Resistance through Loss of Lipopolysaccharide

    OpenAIRE

    García-Quintanilla, Meritxell; Pulido, Marina R.; Moreno-Martínez, Patricia; Martín-Peña, Reyes; López-Rojas, Rafael; Pachón, Jerónimo; McConnell, Michael J

    2014-01-01

    Acinetobacter baumannii can acquire resistance to the cationic peptide antibiotic colistin through complete loss of lipopolysaccharide (LPS) expression. The activities of the host cationic antimicrobials LL-37 and human lysozyme against multidrug-resistant clinical isolates of A. baumannii that acquired colistin resistance through lipopolysaccharide loss were characterized. We demonstrate that LL-37 has activity against strains lacking lipopolysaccharide that is similar to that of their colis...

  13. 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

  14. Lipopolysaccharide density and structure govern the extent and distance of nanoparticle interaction with actual and model bacterial outer membranes

    Energy Technology Data Exchange (ETDEWEB)

    Jacobson, Kurt H.; Gunsolus, Ian L.; Kuech, Thomas R.; Troiano, Julianne M.; Melby, Eric S.; Lohse, Samuel E.; Hu, Dehong; Chrisler, William B.; Murphy, Catherine; Orr, Galya; Geiger, Franz M.; Haynes, Christy L.; Pedersen, Joel A.

    2015-07-24

    Design of nanomedicines and nanoparticle-based antimicrobial and antifouling formulations, and assessment of the potential implications of nanoparticle release into the environment require understanding nanoparticle interaction with bacterial surfaces. Here we demonstrate electrostatically driven association of functionalized nanoparticles with lipopolysaccharides of Gram-negative bacterial outer membranes and find that lipopolysaccharide structure influences the extent and location of binding relative to the lipid-solution interface. By manipulating the lipopolysaccharide content in Shewanella oneidensis outer membranes, we observed electrostatically driven interaction of cationic gold nanoparticles with the lipopolysaccharide-containing leaflet. We probed this interaction by quartz crystal microbalance with dissipation monitoring (QCM-D) and second harmonic generation (SHG) using solid-supported lipopolysaccharide-containing bilayers. Association of cationic nanoparticles increased with lipopolysaccharide content, while no association of anionic nanoparticles was observed. The harmonic-dependence of QCM-D measurements suggested that a population of the cationic nanoparticles was held at a distance from the outer leaflet-solution interface of bilayers containing smooth lipopolysaccharides (those bearing a long O-polysaccharide). Additionally, smooth lipopolysaccharides held the bulk of the associated cationic particles outside of the interfacial zone probed by SHG. Our results demonstrate that positively charged nanoparticles are more likely to interact with Gram-negative bacteria than are negatively charged particles, and this interaction occurs primarily through lipopolysaccharides.

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

    Directory of Open Access Journals (Sweden)

    H. Yang

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

  16. 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

  17. Emodin ameliorates lipopolysaccharides-induced corneal inflammation in rats

    Directory of Open Access Journals (Sweden)

    Guo-Ling Chen

    2015-08-01

    Full Text Available AIM:To investigate the effect of emodin on pseudomonas aeruginosa lipopolysaccharides (LPS-induced corneal inflammation in rats.METHODS:Corneal infection was induced by pseudomonas aeruginosa LPS in Wistar rats. The inflammation induced by LPS were examined by slit lamp microscope and cytological checkup of aqueous humor. Corneal tissue structure was observed by hematoxylin and eosin (HE staining. The activation of nuclear factor kappaB (NF-?B was determined by Western blot. Messenger ribonucleic acid (mRNA of tumor necrosis factor-? (TNF-? and intercellular adhesion molecule-1 (ICAM-1 in LPS-challenged rat corneas were measured with reverse transcription-polymerase chain reaction (RT-PCR.RESULTS:Typical manifestations of acute corneal inflammation were observed in LPS-induce rat model, and the corneal inflammatory response and structure were improved in rats pretreated with emodin. Treatment with emodin could improve corneal structure, reduce corneal injure by reducing corneal inflammatory response. Emodin could inhibit the decreasing lever of inhibitor of kappaB alpha (I?B? express, and the mRNA expression of TNF-? and ICAM-1 in corneal tissues was also inhibited by emodin. The differences were statistically significant between groups treated with emodin and those without treatment (P<0.01.CONCLUSION:Emodin could ameliorate LPS-induced corneal inflammation, which might via inhibiting the activation of NF-?B.

  18. Allergen immunotherapy with nanoparticles containing lipopolysaccharide from Brucella ovis.

    Science.gov (United States)

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

    2008-11-01

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

  19. 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.

  20. Curcumin attenuation of lipopolysaccharide induced cardiac hypertrophy in rodents.

    Science.gov (United States)

    Chowdhury, Rupak; Nimmanapalli, Ramadevi; Graham, Thomas; Reddy, Gopal

    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 cardiomegaly, and weighed to compare heart weight/brain weight (HW/BW) in mg/mg. For immunohistochemistry, the tissue sections were exposed to histone H3, H4 and acetylated histone H3, H4 antibody. LPS induced a significant increase in histone acetylation as shown by intense staining. In curcumin + LPS treated mice nuclear staining was similar to the control group indicating that curcumin traversed the histone acetylation activity of the LPS. To further check the mechanism of action of curcumin, p300 protein acetylation levels were analyzed. This study suggests that the probable mechanism of action of curcumin is via the reduction of p300 HAT activity. PMID:24236240

  1. 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.

  2. Molecular mechanisms in lipopolysaccharide-induced pulmonary endothelial barrier dysfunction.

    Science.gov (United States)

    Liu, Han; Yu, Xiu; Yu, Sulan; Kou, Junping

    2015-12-01

    The confluent pulmonary endothelium plays an important role as a semi-permeable barrier between the vascular space of blood vessels and the underlying tissues, and it contributes to the maintenance of circulatory fluid homeostasis. Pulmonary endothelial barrier dysfunction is a pivotal early step in the development of a variety of high mortality diseases, such as acute lung injury (ALI). Endothelium barrier dysfunction in response to inflammatory or infectious mediators, including lipopolysaccharide (LPS), is accompanied by invertible cell deformation and interendothelial gap formation. However, specific pharmacological therapies aiming at ameliorating pulmonary endothelial barrier function in patients are still lacking. A full understanding of the fundamental mechanisms that are involved in the regulation of pulmonary endothelial permeability is essential for the development of barrier protective therapeutic strategies. Therefore, this review summarizes several important molecular mechanisms involved in LPS-induced changes in pulmonary endothelial barrier function. As for barrier-disruption, the activation of myosin light chain kinase (MLCK), RhoA and tyrosine kinases; increase of calcium influx; and apoptosis of the endothelium lead to an elevation of lung endothelial permeability. Additionally, the activation of Rac1, Cdc42, protease activated receptor 1 (PAR1) and adenosine receptors (ARs), as well as the increase of cyclic AMP and sphingosine-1-phosphate (S1P) content, protect against LPS-induced lung endothelial barrier dysfunction. Furthermore, current regulatory factors and strategies against the development of LPS-induced lung endothelial hyper-permeability are discussed. PMID:26462590

  3. Zingerone attenuates lipopolysaccharide-induced acute lung injury in mice.

    Science.gov (United States)

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

    2014-03-01

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

  4. Ligustrazine effect on lipopolysaccharide-induced pulmonary damage in rats.

    Science.gov (United States)

    Wang, Huiqi; Chen, Yuanzhuo; Li, Wenjie; Li, Congye; Zhang, Xiangyu; Peng, Hu; Gao, Chengjin

    2015-09-01

    We investigated the effectiveness of ligustrazine (tetramethylpyrazine, TMP) in alleviating pulmonary damage induced by lipopolysaccharide (LPS). Twenty-four healthy male Sprague-Dawley rats were randomly divided into three groups: the blank group, LPS group, and TMP treatment group (TMP group). The LPS group was intraperitoneally injected with LPS (20mg/kg), and the TMP group was intraperitoneally injected with LPS (20mg/kg) and ligustrazine (80mg/kg). Blood gas analysis, hematoxylin-eosin staining dye extravasation and diffuse alveolar damage (DAD) score, the wet/dry lung tissue (W/D) ratios, the expression of CD31+ vascular endothelial microparticles (EMPs), tumor necrosis factor alpha (TNF-?) levels, and the protein expression of Rho-associated coiled-coil-forming protein kinase (ROCK) II and Toll-like receptor 4 (TLR4) were analyzed. Compared with the blank group, the arterial partial pressure of oxygen (PaO2) declined in both 1 and 4h (PEMPs, and protein expression of ROCK II and TLR4 were significantly increased (PEMPs, and protein expression of ROCK II and TLR4 were significantly decreased in the TMP group compared with the LPS group (PEMP levels in the plasma, reducing the release of the inflammatory mediator TNF-? and inhibiting the protein expression of ROCK II and TLR4. PMID:26088147

  5. 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

  6. 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

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

    DEFF Research Database (Denmark)

    Stenbaek, E.I.; HovindHaugen, K.

    1996-01-01

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

  8. Inhibition of lipopolysaccharide induced acute inflammation in lung by chlorination.

    Science.gov (United States)

    Zhang, Jinshan; Xue, Jinling; Xu, Bi; Xie, Jiani; Qiao, Juan; Lu, Yun

    2016-02-13

    Lipopolysaccharide (LPS, also called endotoxin) is a pro-inflammatory constituent of gram negative bacteria and cyanobacteria, which causes a potential health risk in the process of routine urban application of reclaimed water, such as car wash, irrigation, scenic water refilling, etc. Previous studies indicated that the common disinfection treatment, chlorination, has little effect on endotoxin activity removal measured by Limulus amebocyte lysate (LAL) assay. However, in this study, significant decrease of acute inflammatory effects was observed in mouse lung, while LAL assay still presented a moderate increase of endotoxin activity. To explore the possible mechanisms, the nuclear magnetic resonance (NMR) results showed the chlorination happened in alkyl chain of LPS molecules, which could affect the interaction between LPS and LPS-binding protein. Also the size of LPS aggregates was found to drop significantly after treatment, which could be another results of chlorination caused polarity change. In conclusion, our observation demonstrated that chlorination is effective to reduce the LPS induced inflammation in lung, and it is recommended to use health effect-based methods to assess risk removal of water treatment technologies. PMID:26530889

  9. Palmitate and lipopolysaccharide trigger synergistic ceramide production in primary macrophages.

    Science.gov (United States)

    Schilling, Joel D; Machkovech, Heather M; He, Li; Sidhu, Rohini; Fujiwara, Hideji; Weber, Kassandra; Ory, Daniel S; Schaffer, Jean E

    2013-02-01

    Macrophages play a key role in host defense and in tissue repair after injury. Emerging evidence suggests that macrophage dysfunction in states of lipid excess can contribute to the development of insulin resistance and may underlie inflammatory complications of diabetes. Ceramides are sphingolipids that modulate a variety of cellular responses including cell death, autophagy, insulin signaling, and inflammation. In this study we investigated the intersection between TLR4-mediated inflammatory signaling and saturated fatty acids with regard to ceramide generation. Primary macrophages treated with lipopolysaccharide (LPS) did not produce C16 ceramide, whereas palmitate exposure led to a modest increase in this sphingolipid. Strikingly, the combination of LPS and palmitate led to a synergistic increase in C16 ceramide. This response occurred via cross-talk at the level of de novo ceramide synthesis in the ER. The synergistic response required TLR4 signaling via MyD88 and TIR-domain-containing adaptor-inducing interferon beta (TRIF), whereas palmitate-induced ceramide production occurred independent of these inflammatory molecules. This ceramide response augmented IL-1? and TNF? release, a process that may contribute to the enhanced inflammatory response in metabolic diseases characterized by dyslipidemia. PMID:23250746

  10. Trapped lipopolysaccharide and LptD intermediates reveal lipopolysaccharide translocation steps across the Escherichia coli outer membrane.

    Science.gov (United States)

    Li, Xuejun; Gu, Yinghong; Dong, Haohao; Wang, Wenjian; Dong, Changjiang

    2015-01-01

    Lipopolysaccharide (LPS) is a main component of the outer membrane of Gram-negative bacteria, which is essential for the vitality of most Gram-negative bacteria and plays a critical role for drug resistance. LptD/E complex forms a N-terminal LPS transport slide, a hydrophobic intramembrane hole and the hydrophilic channel of the barrel, for LPS transport, lipid A insertion and core oligosaccharide and O-antigen polysaccharide translocation, respectively. However, there is no direct evidence to confirm that LptD/E transports LPS from the periplasm to the external leaflet of the outer membrane. By replacing LptD residues with an unnatural amino acid p-benzoyl-L-phenyalanine (pBPA) and UV-photo-cross-linking in E.coli, the translocon and LPS intermediates were obtained at the N-terminal domain, the intramembrane hole, the lumenal gate, the lumen of LptD channel, and the extracellular loop 1 and 4, providing the first direct evidence and "snapshots" to reveal LPS translocation steps across the outer membrane. PMID:26149544

  11. Gram-Negative Bacterial Lipopolysaccharide Stimulates Activin A Secretion from Human Amniotic Epithelial Cells

    Science.gov (United States)

    Marukawa, Risa; Tsuru, Nami; Sato, Maki; Matsuda, Hiroko; Sadakata, Hisanobu; Minegishi, Takashi

    2013-01-01

    Activin A is involved in inflammation. The present study was performed to clarify if lipopolysaccharide, a component of Gram-negative bacteria, stimulates activin A secretion from human amniotic epithelial cells and to determine if activin A plays a role in amnionitis. Fetal membranes were obtained during elective cesarean sections performed in full-term pregnancies of patients without systemic disease, signs of premature delivery, or fetal complications. Amniotic epithelial cells were isolated by trypsinization. The activin A concentrations in the culture media were measured by enzyme-linked immunosorbent assay, and cell proliferation was assessed by 5-bromo-2?-deoxyuridine incorporation. Amniotic epithelial cells secreted activin A in a cell density-dependent manner, and lipopolysaccharide (10??g/mL) enhanced the secretion at each cell density. Lipopolysaccharide (10–50??g/mL) also stimulated activin A secretion in a dose-dependent manner. Contrary to the effect of activin A secretion, lipopolysaccharide inhibited cell proliferation in amniotic epithelial cells. The present study suggests that lipopolysaccharide stimulation of activin A secretion may be a mechanism in the pathogenesis of amnionitis. PMID:23956746

  12. 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.

  13. Peripheral tumors alter neuroinflammatory responses to lipopolysaccharide in female rats.

    Science.gov (United States)

    Pyter, Leah M; El Mouatassim Bih, Sarah; Sattar, Husain; Prendergast, Brian J

    2014-03-13

    Cancer is associated with an increased prevalence of depression. Peripheral tumors induce inflammatory cytokine production in the brain and depressive-like behaviors. Mounting evidence indicates that cytokines are part of a pathway by which peripheral inflammation causes depression. Neuroinflammatory responses to immune challenges can be exacerbated (primed) by prior immunological activation associated with aging, early-life infection, and drug exposure. This experiment tested the hypothesis that peripheral tumors likewise induce neuroinflammatory sensitization or priming. Female rats with chemically-induced mammary carcinomas were injected with either saline or lipopolysaccharide (LPS, 250?g/kg; i.p.), and expression of mRNAs involved in the pathway linking inflammation and depression (interleukin-1beta [Il-1?], CD11b, I?B?, indolamine 2,3-deoxygenase [Ido]) was quantified by qPCR in the hippocampus, hypothalamus, and frontal cortex, 4 or 24h post-treatment. In the absence of LPS, hippocampal Il-1? and CD11b mRNA expression were elevated in tumor-bearing rats, whereas Ido expression was reduced. Moreover, in saline-treated rats basal hypothalamic Il-1? and CD11b expression were positively correlated with tumor weight; heavier tumors, in turn, were characterized by more inflammatory, necrotic, and granulation tissue. Tumors exacerbated CNS proinflammatory gene expression in response to LPS: CD11b was greater in hippocampus and frontal cortex of tumor-bearing relative to tumor-free rats, I?B? was greater in hippocampus, and Ido was greater in hypothalamus. Greater neuroinflammatory responses in tumor-bearing rats were accompanied by attenuated body weight gain post-LPS. The data indicate that neuroinflammatory pathways are potentiated, or primed, in tumor-bearing rats, which may exacerbate future negative behavioral consequences. PMID:24457042

  14. Oxyresveratrol suppresses lipopolysaccharide-induced inflammatory responses in murine macrophages.

    Science.gov (United States)

    Lee, H S; Kim, D H; Hong, J E; Lee, J-Y; Kim, E J

    2015-08-01

    Excessive inflammation is considered a critical factor in many human diseases. Oxyresveratrol(trans-2,3',4,5'-tetrahydroxystilbene), a natural hydroxystilbene, has been shown to possess antioxidant and free radical-scavenging activity. In this study, we investigated the effects of oxyresveratrol (OxyR) on the lipopolysaccharide (LPS)-induced production of inflammatory cytokines and mediators and further explored the mechanism of action in RAW264.7 murine macrophage cell line. Production of nitric oxide (NO), prostaglandin E2 (PGE2), messenger RNA (mRNA) and protein expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin 6 (IL-6), and granulocyte macrophage colony-stimulating factor (GM-CSF), phosphorylation of mitogen-activated protein kinases (MAPKs; extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), p38), and the activation of nuclear factor ?-light chain enhancer of activated B cells (NF?B) with OxyR were assayed in LPS-stimulated RAW264.7 cells. OxyR inhibited the productions of NO, PGE2, IL-6, and GM-CSF significantly in LPS-stimulated RAW264.7 cells. OxyR suppressed mRNA and protein expressions of iNOS, COX-2, IL-6, and GM-CSF in LPS-stimulated RAW264.7 cells. OxyR suppressed the phosphorylation of Akt and JNK and p38 MAPKs and the translocation of NF?B p65 subunit into the nucleus. These results indicate that OxyR inhibits LPS-stimulated inflammatory responses though the blocking of MAPK and NF?B signaling pathway in macrophages, and suggest that OxyR possesses anti-inflammatory effects. PMID:25425548

  15. Altered T Lymphocyte Proliferation upon Lipopolysaccharide Challenge Ex Vivo

    Science.gov (United States)

    Poujol, Fanny; Monneret, Guillaume; Pachot, Alexandre; Textoris, Julien; Venet, Fabienne

    2015-01-01

    Context Sepsis is characterized by the development of adaptive immune cell alterations, which intensity and duration are associated with increased risk of health-care associated infections and mortality. However, pathophysiological mechanisms leading to such lymphocyte dysfunctions are not completely understood, although both intrinsic lymphocyte alterations and antigen-presenting cells (APCs) dysfunctions are most likely involved. Study The aim of the current study was to evaluate whether lipopolysaccharide (LPS, mimicking initial Gram negative bacterial challenge) could directly impact lymphocyte function after sepsis. Therefore, we explored ex-vivo the effect of LPS priming on human T lymphocyte proliferation induced by different stimuli. Results We showed that LPS priming of PBMCs reduced T cell proliferative response and altered IFN? secretion after stimulation with OKT3 but not with phytohaemagglutinin or anti-CD2/CD3/CD28-coated beads stimulations. Interestingly only LPS priming of monocytes led to decreased T cell proliferative response as opposed to LPS priming of lymphocytes. Importantly, LPS priming was associated with reduced expression of HLA-DR, CD86 and CD64 on monocytes but not with the modification of CD3, CTLA4, PD-1 and CD28 expressions on lymphocytes. Finally, IFN? stimulation restored monocytes accessory functions and T cell proliferative response to OKT3. Conclusion We conclude that LPS priming does not directly impact lymphocyte functions but reduces APC’s capacity to activate T cells. This recapitulates ex vivo indirect mechanisms participating in sepsis-induced lymphocyte alterations and suggests that monocyte-targeting immunoadjuvant therapies in sepsis may also help to improve adaptive immune dysfunctions. Direct mechanisms impacting lymphocytes being also at play during sepsis, the respective parts of direct versus indirect sepsis-induced lymphocyte alterations remain to be evaluated in clinic. PMID:26642057

  16. 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.

  17. Soluble CD14 in periodontitis.

    Science.gov (United States)

    Nicu, Elena A; Laine, Marja L; Morré, Servaas A; Van der Velden, Ubele; Loos, Bruno G

    2009-04-01

    Lipopolysaccharide (LPS) binds to soluble (s)CD14. We investigated which factors contribute to variations in sCD14 levels in periodontitis, a chronic infectious disease of tooth-supporting tissues associated with endotoxemia and leading to inflammation and subsequently loss of teeth. The sCD14 levels were determined by ELISA in healthy controls (n=57) and untreated patients (59 moderate and 46 severe) and their relation with markers of systemic inflammation (C-reactive protein levels, and leukocyte, neutrophil and lymphocyte counts) was assessed. Anti-Aggregatibacter actinomycetemcomitans and anti-Porphyromonas gingivalis IgG levels were established by ELISA and CD14(-260) genotype was determined in a TaqMan allelic discrimination assay. Increased levels of sCD14 were more frequent among periodontitis patients (P=0.026) and showed a severity-dependence with increasing levels of periodontal breakdown (P=0.008). In patients, levels of sCD14 correlated positively with CRP (P=0.043), leukocyte numbers (P=0.011) and negatively with anti-A. actinomycetemcomitans IgG (P=0.007). In a multivariate analysis, sCD14 levels were predicted by ethnicity, age, educational level, and in Caucasian subjects also by the severity of periodontal destruction, but not by anti-P. gingivalis IgG or the CD14(-260) genotype. Periodontitis is associated with elevated levels of sCD14. PMID:19318422

  18. Gene expression patterns in bone following lipopolysaccharide stimulation.

    Science.gov (United States)

    Yang, Jing; Su, Nan; Du, Xiaolan; Chen, Lin

    2014-12-01

    Bone displays suppressed osteogenesis in inflammatory diseases such as sepsis and rheumatoid arthritis. However, the underlying mechanisms have not yet been clearly explained. To identify the gene expression patterns in the bone, we performed Affymetrix Mouse Genome 430 2.0 Array with RNA isolated from mouse femurs 4 h after lipopolysaccharide (LPS) administration. The gene expressions were confirmed with real-time PCR. The serum concentration of the N-terminal propeptide of type I collagen (PINP), a bone-formation marker, was determined using ELISA. A total of 1003 transcripts were upregulated and 159 transcripts were downregulated (more than twofold upregulation or downregulation). Increased expression levels of the inflammation-related genes interleukin-6 (IL-6), interleukin-1? (IL-1?) and tumor necrosis factor ? (TNF-?) were confirmed from in the period 4 h to 72 h after LPS administration using real-time PCR. Gene ontogene analysis found four bone-related categories involved in four biological processes: system development, osteoclast differentiation, ossification and bone development. These processes involved 25 upregulated genes. In the KEGG database, we further analyzed the transforming growth factor ? (TGF-?) pathway, which is strongly related to osteogenesis. The upregulated bone morphogenetic protein 2 (BMP2) and downregulated inhibitor of DNA binding 4 (Id4) expressions were further confirmed by real-time PCR after LPS stimulation. The osteoblast function was determined through examination of the expression levels of core binding factor 1 (Cbfa1) and osteocalcin (OC) in bone tissues and serum PINP from 4 h to 72 h after LPS administration. The expressions of OC and Cbfa1 decreased 6 h after administration (p 0.05) of LPS stimulation. The results of this study suggest that LPS induces elevated expressions of skeletal system development- and osteoclast differentiation-related genes and inflammation genes at an early stage in the bone. The perturbed functions of these two groups of genes may lead to a faint change in osteogenesis at an early stage of LPS stimulation. Suppressed bone formation was found at later stages in response to LPS stimulation. PMID:25355239

  19. 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.

  20. PPAR? downregulates airway inflammation induced by lipopolysaccharide in the mouse

    Directory of Open Access Journals (Sweden)

    Frossard Nelly

    2005-08-01

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

  1. Nilotinib ameliorates lipopolysaccharide-induced acute lung injury in rats

    International Nuclear Information System (INIS)

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

  2. Characterization of interleukin-1 receptor antagonist isoform expression in the brain of lipopolysaccharide-treated rats.

    Science.gov (United States)

    Palin, K; Pousset, F; Verrier, D; Dantzer, R; Kelley, K; Parnet, P; Lestage, J

    2001-01-01

    The endogenous interleukin-1 receptor antagonist is the natural inhibitor of the biological effects of interleukin-1 during inflammation. Interleukin-1 receptor antagonist refers to three isoforms: one secreted and two intracellular forms (types I and II). The objective of the present study was to investigate the expression of interleukin-1 receptor antagonist isoforms in the rat brain in vivo in response to an i.p. injection of lipopolysaccharide. The interleukin-1 receptor antagonist was studied at the messenger and protein levels by reverse transcription-polymerase chain reaction and western blot analysis, respectively. Interleukin-1 receptor antagonist messenger RNA was constitutively expressed in the brain and its expression increased in response to lipopolysaccharide. The three interleukin-1 receptor antagonist protein isoforms were up-regulated after lipopolysaccharide treatment in a time-dependent manner. Their relative expression differed according to the isoform and brain region studied. Double immunofluorescence staining revealed interleukin-1 receptor antagonist positive neurons and microglia in hippocampus 24h after lipopolysaccharide stimulation. These results demonstrate for the first time that brain cells are able to produce interleukin-1 receptor antagonist isoforms in response to a peripheral immune challenge with a predominance of the secreted over intracellular forms. PMID:11311797

  3. Bacteriophage K20 requires both the OmpF porin and lipopolysaccharide for receptor function.

    OpenAIRE

    Silverman, J. A.; Benson, S A

    1987-01-01

    Mutations which prevent absorption of the bacteriophage K20 to Escherichia coli K-12 were selected by using an altered OmpF protein which confers the ability to grow on maltodextrin in the absence of the LamB maltoporin. The mutations map in the rfa gene cluster and alter the structure of the lipopolysaccharide core.

  4. Active Immunization with Lipopolysaccharide Pseudomonas Antigen for Chronic Pseudomonas Bronchopneumonia in Guinea Pigs

    OpenAIRE

    Pennington, James E.; Hickey, William F.; Blackwood, Linda L.; Arnaut, M. Amin

    1981-01-01

    Chronic respiratory infection with Pseudomonas aeruginosa is a leading clinical problem among patients with cystic fibrosis. Because antimicrobial agents are usually ineffective in eradicating these infections, additional therapeutic or prophylactic measures should be considered. In this study, an experimental guinea pig model of chronic Pseudomonas aeruginosa bronchopneumonia was utilized to determine whether active immunization with lipopolysaccharide (LPS) P. aeruginosa antigen may favorab...

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

    OpenAIRE

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

    1994-01-01

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

  6. 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

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

    Lifescience Database Archive (English)

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

  8. 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...

  9. Cytokine release by lipopolysaccharide-stimulated whole blood from patients with typhoid fever.

    OpenAIRE

    House, D.; Chinh, NT; Hien, TT; Parry, CP; Ly, NT; Diep, TS; Wain, J.; Dunstan, S; White, NJ; Dougan, G; Farrar, JJ

    2002-01-01

    The ex vivo cytokine response to lipopolysaccharide (LPS) of whole blood from patients with typhoid fever was investigated. Tumor necrosis factor (TNF)-alpha release by LPS-stimulated blood was found to be lower during acute typhoid fever than after a course of antimicrobial therapy (P

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

    Science.gov (United States)

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

  11. 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.

  12. EFFECTS OF CHRONIC EXERCISE CONDITIONING ON THERMAL RESPONSES TO LIPOPOLYSACCHARIDE AND TURPENTINE ABSCESS IN FEMALE RATS.

    Science.gov (United States)

    Chronic exercise conditioning has been shown to alter basal thermoregulatory processes as well as the response to inflammatory agents. Two such agents, lipopolysaccharide (LPS) and turpentine (TPT) are inducers of fever in rats. LPS, given intraperitoneally (i.p.), involves a sys...

  13. 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

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

    OpenAIRE

    Ali S. Salmani; Seyed D. Siadat; Mohammad R. Fallahian; Hojat Ahmadi; Dariushq Norouzian; Parichehr Yaghmai; Mohammad R. Aghasadeghi; Jalal I. Mobarakeh; Seyed M. Sadat; Mehrangize Zangeneh; Maryam Kheirandish

    2009-01-01

    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 smoot...

  15. Demonstration of lipopolysaccharide on sheathed flagella of Vibrio cholerae O:1 by protein A-gold immunoelectron microscopy.

    OpenAIRE

    Fuerst, J A; Perry, J W

    1988-01-01

    Monoclonal antibodies with group and type specificity for lipopolysaccharide antigens were used in combination with protein A-colloidal gold labeling and transmission electron microscopy to demonstrate the presence of lipopolysaccharide antigens on both the sheathed flagellum and the cell surface of Inaba and Ogawa strains of Vibrio cholerae O:1. Labeling was associated with the sheath of the flagellum rather than the core, and flagellar cores were not labeled. Flagellum and cell shared a com...

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

    Science.gov (United States)

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

    1997-02-01

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

  17. 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.

  18. Inhibitory activity of plant stilbenoids against nitric oxide production by lipopolysaccharide-activated microglia.

    Science.gov (United States)

    Nassra, Merian; Krisa, Stéphanie; Papastamoulis, Yorgos; Kapche, Gilbert Deccaux; Bisson, Jonathan; André, Caroline; Konsman, Jan-Pieter; Schmitter, Jean-Marie; Mérillon, Jean-Michel; Waffo-Téguo, Pierre

    2013-07-01

    Microglia-driven inflammatory processes are thought to play an important role in ageing and several neurological disorders. Since consumption of a diet rich in polyphenols has been associated with anti-inflammatory and neuroprotective effects, we studied the effects of twenty-five stilbenoids isolated from Milicia excelsa, Morus alba, Gnetum africanum, and Vitis vinifera. These compounds were tested at 5 and 10 µM on BV-2 microglial cells stimulated with bacterial lipopolysaccharide. Ten stilbenoids reduced lipopolysaccharide-induced nitric oxide production at 5 and/or 10 µM. Two tetramers, E-vitisin A and E-vitisin B, were the most effective molecules. Moreover, they attenuated the expression of the inducible NO synthase protein and gene. PMID:23807809

  19. 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...

  20. Physical contact between lipopolysaccharide and Toll-like receptor 4 revealed by genetic complementation

    OpenAIRE

    Poltorak, A.; Ricciardi-Castagnoli, P.; Citterio, S.; Beutler, B.

    2000-01-01

    Some mammalian species show an ability to discriminate between different lipopolysaccharide (LPS) partial structures (for example, lipid A and its congener LA-14-PP, which lacks secondary acyl chains), whereas others do not. Using a novel genetic complementation system involving the transduction of immortalized macrophages from genetically unresponsive C3H/HeJ mice, we now have shown that the species-dependent discrimination between intact LPS and tetra-acyl LPS partial structures is fully at...

  1. The Lipopolysaccharide Core of Brucella abortus Acts as a Shield Against Innate Immunity Recognition

    OpenAIRE

    Conde Álvarez, R.; Grilló, María Jesús; Llobet Brossa, Enrique; Bengoechea, José Antonio; Gorvel, Jean P

    2012-01-01

    Innate immunity recognizes bacterial molecules bearing pathogen-associated molecular patterns to launch inflammatory responses leading to the activation of adaptive immunity. However, the lipopolysaccharide (LPS) of the gram-negative bacterium Brucella lacks a marked pathogen-associated molecular pattern, and it has been postulated that this delays the development of immunity, creating a gap that is critical for the bacterium to reach the intracellular replicative niche. We found that a B. ab...

  2. Effect of Brucella abortus lipopolysaccharide on oxidative metabolism and lysozyme release by human neutrophils.

    OpenAIRE

    Rasool, O; Freer, E; Moreno, E.; Jarstrand, C.

    1992-01-01

    Both Brucella abortus lipopolysaccharide (LPS) and lipid A were low activators of nitroblue tetrazolium reduction and lysozyme release in human neutrophils. The stimulation was dose dependent and was higher in the presence of autologous plasma than in its absence. The comparison between Brucella LPS and lipid A versus Salmonella LPS revealed that at least 100 times more LPS and 1,000 times more lipid A of the former genus were required to induce significant nitroblue tetrazolium reduction and...

  3. Interaction of Brucella abortus Lipopolysaccharide with Major Histocompatibility Complex Class II Molecules in B Lymphocytes

    OpenAIRE

    Forestier, Claire; Moreno, Edgardo; Méresse, Stéphane; Phalipon, Armelle; Olive, Daniel; Sansonetti, Philippe; Gorvel, Jean-Pierre

    1999-01-01

    Lipopolysaccharide (LPS), a major amphiphilic molecule located at the outer membrane of gram-negative bacteria, is a potent antigen known to induce specific humoral immune responses in infected mammals. LPS has been described as a polyclonal activator of B lymphocytes, triggering the secretion of antibodies directed against distinct sugar epitopes of the LPS chain. But, how LPS is handled by B cells remains to be fully understood. This task appears to be essential for a better knowledge of th...

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

    OpenAIRE

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

    2012-01-01

    Brucella abortus lipopolysaccharide (LPS) was isolated and purified from rough (RB51) and smooth (S2308) strains of Brucella. The LPS preparations were used to treat murine (RAW 264.7) macrophages in order to study their differential effects. Treated macrophages were tested by lysozyme release test (LRT), nitroblue tetrazolium test (NBT) and nitric oxide (NO) assay, respectively. Rough Brucella LPS induced significantly higher levels of lysozyme release, oxidative stress, and nitric oxide in...

  5. Lipopolysaccharide Heterogeneity in the Atypical Group of Novel Emerging Brucella Species

    OpenAIRE

    Zygmunt, Michel S; Jacques, Isabelle; Bernardet, Nelly; Cloeckaert, Axel

    2012-01-01

    Recently, novel Brucella strains with phenotypic characteristics that were atypical for strains belonging to the genus Brucella have been reported. Phenotypically many of these strains were initially misidentified as Ochrobactrum spp. Two novel species have been described so far for these strains, i.e., B. microti and B. inopinata, and other strains genetically related to B. inopinata may constitute other novel species as well. In this study, we analyzed the lipopolysaccharides (LPS) (smooth ...

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

    OpenAIRE

    Kavoosi G; Kabodanian Ardestani S; Kariminia A

    2001-01-01

    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 again...

  7. Production and characterization of monoclonal antibodies specific for the lipopolysaccharide of Escherichia coli O157.

    OpenAIRE

    Westerman, R B; Y He; Keen, J E; Littledike, E. T.; Kwang, J.

    1997-01-01

    Identification of the O157 antigen is an essential part of the detection of Escherichia coli O157:H7, which is recognized as a major etiologic agent of hemorrhagic colitis. However, polyclonal antibodies produced against E. coli O157:H7 lipopolysaccharide (LPS) may react with several other bacteria including Brucella abortus, Brucella melitensis, Yersinia enterocolitica O9, Escherichia hermannii, and Stenotrophomonas maltophilia. We produced eight monoclonal antibodies (MAbs) specific for the...

  8. Analysis of Brucella lipopolysaccharide with specific and cross-reacting monoclonal antibodies.

    OpenAIRE

    Palmer, D. A.; Douglas, J. T.

    1989-01-01

    Monoclonal antibodies which bind Brucella A lipopolysaccharide (LPS)-specific, M LPS-specific, or cross-reactive epitopes were used as reagents in quantitative dot blot, Western blot (immunoblot), and immunoprecipitation analysis of Brucella whole cells, whole-cell extracts, and purified LPS preparations. This set of monoclonal antibodies detected four unique epitopes on Brucella LPS. The specificity of monoclonal antibodies reactive with Brucella unique (A and M) and common (C and C/Y) LPS e...

  9. Striking Complexity of Lipopolysaccharide Defects in a Collection of Sinorhizobium meliloti Mutants

    OpenAIRE

    Campbell, Gordon R. O.; Sharypova, Larissa A.; Scheidle, Heiko; Kathryn M. Jones; Niehaus, Karsten; Becker, Anke; Walker, Graham C.

    2003-01-01

    Although the role that lipopolysaccharide (LPS) plays in the symbiosis between Sinorhizobium meliloti and alfalfa has been studied for over a decade, its function in this process remains controversial and poorly understood. This is largely due to a lack of mutants affected by its synthesis. In one of the definitive studies concerning this issue, Clover et al. (R. H. Clover, J. Kieber, and E. R. Signer, J. Bacteriol. 171:3961-3967, 1989) identified a series of mutants with putative LPS defects...

  10. Beryllium Alters Lipopolysaccharide-Mediated Intracellular Phosphorylation and Cytokine Release in Human Peripheral Blood Mononuclear Cells

    OpenAIRE

    Silva, Shannon; Ganguly, Kumkum; Fresquez, Theresa M.; Gupta, Goutam; McCleskey, T. Mark; Chaudhary, Anu

    2009-01-01

    Beryllium exposure in susceptible individuals leads to the development of chronic beryllium disease, a lung disorder marked by release of inflammatory cytokine and granuloma formation. We have previously reported that beryllium induces an immune response even in blood mononuclear cells from healthy individuals. In this study, we investigate the effects of beryllium on lipopolysaccharide - mediated cytokine release in blood mononuclear and dendritic cells from healthy individuals. We find that...

  11. Uncoupling protein 2 plays an important role in nitric oxide production of lipopolysaccharide-stimulated macrophages

    OpenAIRE

    Kizaki, Takako; SUZUKI, Kenji; HITOMI, YOSHIAKI; Taniguchi, Naoyuki; Saitoh, Daizoh; Watanabe, Kenji; Onoé, Kazunori; Day, Noorbibi K.; Good, Robert A.; Ohno, Hideki

    2002-01-01

    The expression of uncoupling protein 2 (UCP2) was reduced in macrophages after stimulation with lipopolysaccharide (LPS). The physiological consequence and the regulatory mechanisms of the UCP2 down-regulation by LPS were investigated in a macrophage cell line, RAW264 cells. UCP2 overexpression in RAW264 cells transfected with eukaryotic expression vector containing ucp2 cDNA markedly reduced the production of intracellular reactive oxygen species. Furthermore, in the UCP...

  12. Longitudinal study of antibody response to lipopolysaccharides during chronic Pseudomonas aeruginosa lung infection in cystic fibrosis.

    OpenAIRE

    Fomsgaard, A.; Høiby, N.; Shand, G H; Conrad, R S; Galanos, C

    1988-01-01

    Antibodies to Pseudomonas aeruginosa from 10 cystic fibrosis patients with chronic P. aeruginosa lung infections were quantitatively and qualitatively analyzed. The development of specific antibodies in patient serum was evaluated in a longitudinal study (1972 to 1987). The concentrations and specificities of immunoglobulin G (IgG) and IgM antibodies to purified lipopolysaccharides (LPS) from clinical isolates of P. aeruginosa and to a variety of other gram-negative bacteria were studied by i...

  13. Inhibitory Effect of Gallic acid on Production of Interleukins in Mouse Macrophage Stimulated by Lipopolysaccharide

    OpenAIRE

    Wansu Park

    2010-01-01

    Objectives: Gallic acid (GA) is the major component of tannin which could be easily founded in various natural materials such as green tea, red tea, grape juice, and Corni Fructus. The purpose of this study is to investigate the effect of Gallic acid (GA) on production of interleukin (IL) in mouse macrophage Raw 264.7 cells stimulated by lipopolysaccharide (LPS). Methods: Productions of interleukins were measured by High-throughput Multiplex Bead based Assay with Bio-plex Suspension Array ...

  14. Bacterial lipopolysaccharide-induced intestinal microvascular lesions leading to acute diarrhea.

    OpenAIRE

    Mathan, V I; Penny, G R; Mathan, M M; Rowley, D

    1988-01-01

    Subcutaneous challenge of mice with lipopolysaccharide (LPS) from gram negative bacteria, produced an intestinal microvascular lesion causing fluid exudation into the lumen of the intestine and diarrhea. The microvascular lesion was characterized by endothelial cell damage and microthrombi in the venules and capillaries of the intestinal lamina propria. Marker organisms, given orally to challenged mice, grew in the exuded fluid and could invade the mucosa. Intravenous transfer of postchalleng...

  15. Heat shock inhibits lipopolysaccharide-induced tissue factor activity in human whole blood

    OpenAIRE

    Thielmann Matthias; Zacharowski Kai; Sucker Christoph; Hartmann Matthias

    2007-01-01

    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 bl...

  16. Lipopolysaccharide-Induced Neuronal Activation in the Paraventricular and Dorsomedial Hypothalamus Depends on Ambient Temperature

    OpenAIRE

    Wanner, Samuel P; Yoshida, Kyoko; Vladimir A. Kulchitsky; Ivanov, Andrei I; Kanosue, Kazuyuki; Romanovsky, Andrej A.

    2013-01-01

    Systemic inflammatory response syndrome is associated with either fever or hypothermia, but the mechanisms responsible for switching from one to the other are unknown. In experimental animals, systemic inflammation is often induced by bacterial lipopolysaccharide (LPS). To identify the diencephalic and brainstem structures involved in the fever-hypothermia switch, we studied the expression of c-Fos protein, a marker of neuronal activation, in rats treated with the same high dose of LPS (0.5 m...

  17. Gram-Negative Bacterial Lipopolysaccharide Stimulates Activin A Secretion from Human Amniotic Epithelial Cells

    OpenAIRE

    Takashi Kameda; Takashi Minegishi; Hisanobu Sadakata; Hiroko Matsuda; Risa Marukawa; Nami Tsuru; Maki Sato; Yumiko Abe

    2013-01-01

    Activin A is involved in inflammation. The present study was performed to clarify if lipopolysaccharide, a component of Gram-negative bacteria, stimulates activin A secretion from human amniotic epithelial cells and to determine if activin A plays a role in amnionitis. Fetal membranes were obtained during elective cesarean sections performed in full-term pregnancies of patients without systemic disease, signs of premature delivery, or fetal complications. Amniotic epithelial cells were isolat...

  18. Human platelet aggregation is initiated by peripheral blood mononuclear cells exposed to bacterial lipopolysaccharide in vitro.

    OpenAIRE

    Schwartz, B. S.; Monroe, M. C.

    1986-01-01

    Platelet consumption is a prominent feature of disseminated intravascular coagulation. We investigated whether monocyte procoagulant activity (PCA) might play a role in platelet consumption associated with gram-negative septicemia. Human mononuclear cells exposed in vitro to lipopolysaccharide demonstrated parallel dose-dependent increases in PCA and ability to induce platelet aggregation. Induction of platelet aggregation required the generation of thrombin dependent on coagulation Factors V...

  19. The role of lipopolysaccharide binding protein in innate immunity in infected partial thickness burns

    OpenAIRE

    Lahoda, Lars-Uwe,

    2012-01-01

    Wanneer in een brandwond infectie optreedt, neemt het risico op ziekte en overlijden van de patiënt sterk toe. Vooral infecties met gramnegatieve bacteriën zijn gevaarlijk. Over hoe bacteriën het aangeboren immuunsysteem in een (oppervlakkige) brandwond beïnvloeden, bestaat nog veel onduidelijkheid. Lars-Uwe Lahoda verrichtte onderzoek op dit vlak. Zijn onderzoek laat zien dat het eiwit lipopolysaccharide binding protein (LBP) een belangrijke rol speelt bij de bestrijding van infecties in...

  20. Rhizobium meliloti lipopolysaccharide and exopolysaccharide can have the same function in the plant-bacterium interaction.

    OpenAIRE

    Putnoky, P; Petrovics, G; Kereszt, A; Grosskopf, E; Ha, D T; Bánfalvi, Z; Kondorosi, A

    1990-01-01

    A fix region of Rhizobium meliloti 41 involved both in symbiotic nodule development and in the adsorption of bacteriophage 16-3 was delimited by directed Tn5 mutagenesis. Mutations in this DNA region were assigned to four complementation units and were mapped close to the pyr-2 and pyr-29 chromosomal markers. Phage inactivation studies with bacterial cell envelope preparations and crude lipopolysaccharides (LPS) as well as preliminary characterization of LPS in the mutants indicated that thes...

  1. 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...

  2. Copper chelation by tetrathiomolybdate inhibits lipopolysaccharide-induced inflammatory responses in vivo

    OpenAIRE

    Wei, Hao; Frei, Balz; Beckman, Joseph S; Zhang, Wei-jian

    2011-01-01

    Redox-active transition metal ions, such as iron and copper, may play an important role in vascular inflammation, which is an etiologic factor in atherosclerotic vascular diseases. In this study, we investigated whether tetrathiomolybdate (TTM), a highly specific copper chelator, can act as an anti-inflammatory agent, preventing lipopolysaccharide (LPS)-induced inflammatory responses in vivo. Female C57BL/6N mice were daily gavaged with TTM (30 mg/kg body wt) or vehicle control. After 3 wk, a...

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

    OpenAIRE

    Liang-Takasaki, C J; Saxén, H; Mäkelä, P.H.; Leive, L

    1983-01-01

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

  4. Immunization with major outer membrane protein (porin) preparations in experimental murine salmonellosis: effect of lipopolysaccharide.

    OpenAIRE

    Kuusi, N; Nurminen, M; Saxén, H; Mäkelä, P.H.

    1981-01-01

    A crude major outer membrane (porin) preparation, obtained from a rough strain of Salmonella typhimurium earlier shown to be protective both in active and passive immunization of mice against challenge with smooth S. typhimurium, was further purified. Removal of the main impurities, lipopolysaccharide (LPS) and lipoprotein, was accompanied by loss of protective capacity in passive immunization experiments. Reconstitution with rough LPS restored the protective capacity. Protection was, however...

  5. Activation of nitric oxide synthase and induction of defense genes in Arabidopsis thaliana by bacterial lipopolysaccharides

    OpenAIRE

    Zeidler, Dana

    2006-01-01

    The aim of this study was to examine if Lipopolysaccharide (LPS) are novel elicitors of plant innate immunity using Arabidopsis thaliana as a model system. LPS are the major outer membrane components of Gram-negative bacteria and consist of three distinct structural domains: O-antigen, core region and lipid A. They represent microbe-/pathogen-associated molecular patterns (PAMPs) in animal patho-systems and act as extremely potent stimulators of the mammalian and insect innate immunity. As fo...

  6. A serine protease zymogen functions as a pattern-recognition receptor for lipopolysaccharides

    OpenAIRE

    Ariki, Shigeru; Koori, Kumiko; Osaki, Tsukasa; Motoyama, Kiyohito; Inamori, Kei-ichiro; Kawabata, Shun-Ichiro

    2004-01-01

    Bacterial lipopolysaccharide (LPS)-induced exocytosis of granular hemocytes is a key component of the horseshoe crab's innate immunity to infectious microorganisms; stimulation by LPS induces the secretion of various defense molecules from the granular hemocytes. Using a previously uncharacterized assay for exocytosis, we clearly show that hemocytes respond only to LPS and not to other pathogen-associated molecular patterns, such as ?-1,3-glucans and peptidoglycans. Furthermore, we show that ...

  7. Alkaline Phosphatase Protects Lipopolysaccharide-Induced Early Pregnancy Defects in Mice

    OpenAIRE

    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 ro...

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

    OpenAIRE

    Maria G?owacka; Agnieszka St?pie?; Sylwia Szyprowska

    1996-01-01

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

  9. Arginine-Rich Cationic Polypeptides Amplify Lipopolysaccharide-Induced Monocyte Activation

    OpenAIRE

    Bosshart, Herbert; Heinzelmann, Michael

    2002-01-01

    The human neutrophil-derived cationic protein CAP37, also known as azurocidin or heparin-binding protein, enhances the lipopolysaccharide (LPS)-induced release of tumor necrosis factor alpha (TNF-?) in isolated human monocytes. We measured the release of the proinflammatory cytokine interleukin-8 (IL-8) in human whole blood and found that in addition to CAP37, other arginine-rich cationic polypeptides, such as the small structurally related protamines, enhance LPS-induced monocyte activation....

  10. 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...

  11. Development of Immunochromatography-Based Methods for Detection of Leptospiral Lipopolysaccharide Antigen in Urine

    OpenAIRE

    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

    2013-01-01

    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...

  12. Inhibition of Salmonella enterica Serovar Typhimurium Lipopolysaccharide Deacylation by Aminoarabinose Membrane Modification

    OpenAIRE

    Kawasaki, Kiyoshi; Ernst, Robert K; Miller, Samuel I.

    2005-01-01

    Salmonella enterica serovar Typhimurium remodels the lipid A component of lipopolysaccharide, a major component of the outer membrane, to survive within animals. The activation of the sensor kinase PhoQ in host environments increases the synthesis of enzymes that deacylate, palmitoylate, hydroxylate, and attach aminoarabinose to lipid A, also known as endotoxin. These modifications promote bacterial resistance to antimicrobial peptides and reduce the host recognition of lipid A by Toll-like r...

  13. Rhizobium lipopolysaccharide modulates infection thread development in white clover root hairs.

    OpenAIRE

    Dazzo, F. B.; Truchet, G L; Hollingsworth, R. I.; Hrabak, E M; Pankratz, H. S.; Philip-Hollingsworth, S; Salzwedel, J L; Chapman, K; Appenzeller, L; Squartini, A

    1991-01-01

    The interaction between Rhizobium lipopolysaccharide (LPS) and white clover roots was examined. The Limulus lysate assay indicated that Rhizobium leguminosarum bv. trifolii (hereafter called R. trifolii) released LPS into the external root environment of slide cultures. Immunofluorescence and immunoelectron microscopy showed that purified LPS from R. trifolii 0403 bound rapidly to root hair tips and infiltrated across the root hair wall. Infection thread formation in root hairs was promoted b...

  14. 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.; Barkholt, Vibeke; Frøkiær, Hanne

    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 p...

  15. 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 ...

  16. Upregulating Nonneuronal Cholinergic Activity Decreases TNF Release from Lipopolysaccharide-Stimulated RAW264.7 Cells

    OpenAIRE

    Yi Lv; Sen Hu; Jiangyang Lu; Ning Dong; Qian Liu; Minghua Du; Huiping Zhang

    2014-01-01

    Nonneuronal cholinergic system plays a primary role in maintaining homeostasis. It has been proved that endogenous neuronal acetylcholine (ACh) could play an anti-inflammatory role, and exogenous cholinergic agonists could weaken macrophages inflammatory response to lipopolysaccharide (LPS) stimulation through activation of ?7 subunit-containing nicotinic acetylcholine receptor (?7nAChR). We assumed that nonneuronal cholinergic system existing in macrophages could modulate inflammation throug...

  17. Structural Correlation Between Lipophilicity and Lipopolysaccharide-sequestering activity in Spermine-Sulfonamide Analogs

    OpenAIRE

    Burns, Mark R.; Jenkins, Scott A.; Vermeulen, Nicolas M.; Balakrishna, Rajalakshmi; Nguyen, Thuan B.; Kimbrell, Matthew R.; David, Sunil A.

    2006-01-01

    Lipopolysaccharides (LPS), otherwise termed ‘endotoxins’, are outer-membrane constituents of Gram-negative bacteria, and play a key role in the pathogenesis of ‘Septic Shock’, a major cause of mortality in the critically ill patient. We had previously defined the pharmacophore necessary for small molecules to specifically bind and neutralize this complex carbohydrate. A series of aryl and aliphatic spermine-sulfonamide analogs were synthesized and tested in a series of binding and cell-based ...

  18. Gamma interferon cooperates with lipopolysaccharide to activate mouse splenic macrophages to an antihistoplasma state.

    OpenAIRE

    Lane, T E; Wu-Hsieh, B A; Howard, D H

    1993-01-01

    Inhibition of the intracellular growth of Histoplasma capsulatum by murine resident red pulp splenic macrophages was examined. Splenic macrophages, unlike resident peritoneal macrophages, required a prolonged preincubation (18 h) with recombinant murine gamma interferon (rMuIFN-gamma) for activation. To be fully activated, the splenic macrophages required incubation with rMuIFN-gamma in combination with 0.1 microgram of lipopolysaccharide (LPS) per ml. Splenic macrophages stimulated with rMuI...

  19. Role of lipopolysaccharide and complement in susceptibility of Klebsiella pneumoniae to nonimmune serum.

    OpenAIRE

    Ciurana, B; Tomás, J M

    1987-01-01

    The role of lipopolysaccharide (LPS) in the susceptibility of Klebsiella pneumoniae to serum and the mechanism of complement activation by serum-susceptible (SerS) strains were investigated. The classical and alternative complement pathways are involved in serum killing of susceptible K. pneumoniae strains. The LPS composition seems to play a very important role in the serum bactericidal reaction, while capsular polysaccharide from this bacterium does not play any role. High-molecular-weight ...

  20. Enzyme-linked immunosorbent assay for detection of Salmonella lipopolysaccharide in poultry specimens.

    OpenAIRE

    Rigby, C E

    1984-01-01

    An enzyme-linked immunosorbent assay (ELISA) for detection of salmonellae was developed and evaluated by using artificially contaminated specimens of poultry feed, feces, litter, or carcass rinsings, and naturally contaminated water samples. Specimens containing salmonellae of serogroups B or C2 inhibited the binding of polyvalent anti-O serum to microtiter plate wells coated with lipopolysaccharide of Salmonella typhimurium (serogroup B) or Salmonella albany (serogroup C2), respectively. Tre...

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

    OpenAIRE

    Hall, C. L.; Munford, R S

    1983-01-01

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

  2. Redox Imbalance Differentially Inhibits Lipopolysaccharide-Induced Macrophage Activation in the Mouse Liver

    OpenAIRE

    Wang, Fuan; Wang, Luke Y.; Wright, Douglas; Parmely, Michael J.

    1999-01-01

    Endotoxemia is accompanied by significant changes in the reductive-oxidative (redox) balance of critical target organs. Redox stress has been shown to regulate the expression of proinflammatory genes that are induced by endotoxic lipopolysaccharide (LPS) in vitro; however, much less is known about the effects of redox imbalance on LPS-induced gene expression in vivo. To assess the effects of redox stress on inflammatory responses in endotoxemia, mice were treated with either diethyl maleate (...

  3. Ingestion of bacterial lipopolysaccharide inhibits peripheral taste responses to sucrose in mice

    OpenAIRE

    Zhu, Xiaobin; He, Lianying; McCluskey, Lynnette Phillips

    2013-01-01

    A fundamental role of the taste system is to discriminate between nutritive and toxic foods. However, it is unknown whether bacterial pathogens that might contaminate food and water modulate the transmission of taste input to the brain. We hypothesized that exogenous, bacterially-derived lipopolysaccharide (LPS), modulates neural responses to taste stimuli. Neurophysiological responses from the chorda tympani nerve, which innervates taste cells on the anterior tongue, were unchanged by acute ...

  4. Genetic Characterization of the Klebsiella pneumoniae waa Gene Cluster, Involved in Core Lipopolysaccharide Biosynthesis

    OpenAIRE

    Regué, Miguel; Climent, Núria; Abitiu, Nihal; Coderch, Núria; Merino, Susana; Izquierdo, Luis; Altarriba, Maria; Tomás, Juan M.

    2001-01-01

    A recombinant cosmid containing genes involved in Klebsiella pneumoniae C3 core lipopolysaccharide biosynthesis was identified by its ability to confer bacteriocin 28b resistance to Escherichia coli K-12. The recombinant cosmid contains 12 genes, the whole waa gene cluster, flanked by kbl and coaD genes, as was found in E. coli K-12. PCR amplification analysis showed that this cluster is conserved in representative K. pneumoniae strains. Partial nucleotide sequence determination showed that t...

  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 ± 3.4 nm) 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. 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.

  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. An in-vitro evaluation of the efficacy of garlic extract as an antimicrobial agent on periodontal pathogens: A microbiological study.

    Science.gov (United States)

    Shetty, Sunaina; Thomas, Biju; Shetty, Veena; Bhandary, Rahul; Shetty, Raghavendra M

    2013-10-01

    With the rise in bacterial resistance to antibiotics, there is considerable interest in the development of other classes of antimicrobials for the control of infection. Garlic (Allium sativum Linn.) has been used as medicine since ancient times and has long been known to have antibacterial, antifungal, and antiviral properties. This study was undertaken to assess the inhibitory effect of garlic on Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans, to assess the time-kill curve of P. gingivalis and A. actinomycetemcomitans, and to determine the antiproteolytic activity of garlic on P. gingivalis. Ethanolic garlic extract (EGE) and aqueous garlic extract (AGE) were prepared and the inhibitory effects of these extracts for two periodontal pathogens (P. gingivalis and A. actinomycetemcomitans) were tested. Antiproteolytic activity on protease of P. gingivalis was determined. 25 microliter (?l), 50 ?l, and 75 ?l of AGE showed 16 mm, 20 mm, and 25 mm zone of inhibition, respectively, on P. gingivalis. The AGE showed greater bacteriostatic activity against the P. gingivalis with minimum inhibitory concentration determined at 16.6 ?l/ml. The time-kill assay of AGE and EGE were compared for P. gingivalis and A. actinomycetemcomitans. AGE showed better antiproteolytic activity on total protease of P. gingivalis compared to the EGE. Thus, the study concludes the antimicrobial activity of garlic extract against periodontal pathogens, P. gingivalis, A. actinomycetemcomitans. Its action against P. gingivalis includes inhibition of total protease activity, and this raises the possibility that garlic may have therapeutic use for periodontitis and possibly other oral infections. PMID:24695825

  9. Mutation of the Lipopolysaccharide Core Glycosyltransferase Encoded by waaG Destabilizes the Outer Membrane of Escherichia coli by Interfering with Core Phosphorylation

    OpenAIRE

    Yethon, Jeremy A.; Vinogradov, Evgeny; Perry, Malcolm B.; Whitfield, Chris

    2000-01-01

    In Escherichia coli, phosphoryl substituents in the lipopolysaccharide core region are essential for outer membrane stability. Mutation of the core glucosyltransferase encoded by waaG (formerly rfaG) resulted in lipopolysaccharide truncated immediately after the inner core heptose residues, which serve as the sites for phosphorylation. Surprisingly, mutation of waaG also destabilized the outer membrane. Structural analyses of waaG mutant lipopolysaccharide showed that the cause for this pheno...

  10. 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

  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. Induction of IL-10-producing CD4+ T-cells in Chronic Periodontitis

    OpenAIRE

    Kobayashi, R.; Kono, T.(KEK, High Energy Accelerator Research Organization, Tsukuba, Japan); Bolerjack, B.A.; Fukuyama, Y.; Gilbert, R.S.; Ruby, J,; Kataoka, K.; Wada, M; Yamamoto, M; Fujihashi, K.

    2011-01-01

    Precise immunological aspects of inflamed gingival mucosa remain to be elucidated in the murine experimental periodontitis model; therefore, we have characterized the mucosal immune cells in the inflamed gingiva of mice with alveolar bone reduction. Mice were orally infected with Porphyromonas gingivalis 15 times over 2 weeks. Gingival mononuclear cells (GMCs) were isolated from P. gingivalis- and sham-infected mice 1, 7, 15, and 30 days after the last infection. Although the greatest degree ...

  13. Bio-inspired stable antimicrobial peptide coatings for dental applications

    OpenAIRE

    Holmberg, Kyle V.; Abdolhosseini, Mahsa; Li, Yuping; CHEN, XI; Gorr, Sven-Ulrik; Aparicio, Conrado

    2013-01-01

    We developed a novel titanium coating that has applications for preventing infection-related implant failures in dentistry and orthopedics. The coating incorporates an antimicrobial peptide, GL13K, derived from parotid secretory protein, which has been previously shown to be bactericidal and bacteriostatic in solution. We characterized the resulting physicochemical properties, resistance to degradation, activity against Porphyromonas gingivalis, and in vitro cytocompatibility. P. gingivalis i...

  14. 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.

  15. The role of lipopolysaccharide/toll-like receptor 4 signaling in chronic liver diseases

    OpenAIRE

    Soares, João-Bruno; Pimentel-Nunes, Pedro; Roncon-Albuquerque, Roberto; Leite-Moreira, Adelino

    2010-01-01

    Toll-like receptor 4 (TLR4) is a pattern recognition receptor that functions as lipopolysaccharide (LPS) sensor and whose activation results in the production of several pro-inflammatory, antiviral, and anti-bacterial cytokines. TLR4 is expressed in several cells of healthy liver. Despite the constant confrontation of hepatic TLR4 with gut-derived LPS, the normal liver does not show signs of inflammation due to its low expression of TLR4 and ability to modulate TLR4 signaling. Nevertheless, t...

  16. Enzyme-linked immunosorbent assay with Brucella native hapten polysaccharide and smooth lipopolysaccharide.

    OpenAIRE

    Alonso-Urmeneta, B.; Moriyón, I; Díaz, R.; Blasco, J. M.

    1988-01-01

    Brucella melitensis native haptens (NH) are polysaccharides identical to the O-side chain of the smooth lipopolysaccharide (S-LPS) (E. Moreno, H. Mayer, and I. Moriyón, Infect. Immun. 55:2850-2853, 1987) which precipitate with sera from infected cattle but not from strain 19-vaccinated cattle. In the present work, NH was extracted by the hot-water method (R. Díaz, J. Toyos, M.D. Salvo, and M.L. Pardo, Ann. Rech. Vet. 12:35-39, 1981) and purified free of S-LPS and protein. Purified NH lacked t...

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

    OpenAIRE

    López-Urrutia, Luis; Alonso, Andrés; Nieto, Maria Luisa; Bayón, Yolanda; Orduña, Antonio; Sánchez 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...

  18. Induction of immune and adjuvant immunoglobulin G responses in mice by Brucella lipopolysaccharide.

    OpenAIRE

    Moreno, E.; Kurtz, R S; Berman, D. T.

    1984-01-01

    The immunogenic and adjuvant properties of Brucella abortus and Escherichia coli lipopolysaccharides (LPSs) were studied in endotoxin-responsive, athymic, and euthymic BALB/c mice and in responsive C3H/HeAu mice and congenic nonresponsive C3H/HeJ mice. Consistent with previous reports, E. coli LPS did not stimulate significant primary or secondary antibody responses in C3H/HeJ mice and induced the production of immunoglobulin M (IgM) and low levels of IgG in C3H/HeAu mice. In contrast, B. abo...

  19. Structure of the polysaccharides from the lipopolysaccharide of Azospirillum brasilense Jm125A2.

    Science.gov (United States)

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

    2015-10-30

    Two polysaccharides were obtained by mild acid degradation of the lipopolysaccharide of associative nitrogen-fixing bacteria Azospirillum brasilense Jm125A2 isolated from the rhizosphere of a pearl millet. The following structures of the polysaccharides were established by sugar and methylation analyses, Smith degradation, and (1)H and (13)C NMR spectroscopy: Structure 1 has been reported earlier for a polysaccharide from A.?brasilense S17 (Fedonenko YP, Konnova ON, Zdorovenko EL, Konnova SA, Zatonsky GV, Shaskov AS, Ignatov VV, Knirel YA. Carbohydr Res 2008;343:810-6), whereas to our knowledge structure 2 has not been hitherto found in bacterial polysaccharides. PMID:26343325

  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. Lipopolysaccharide Inhibits the Channel Activity of the P2X7 Receptor

    OpenAIRE

    Alejandro Escobar; Claudio Acuña-Castillo; J. Pablo Huidobro-Toro; Margarita Montoya; Jorge Escobar; Miguel Rios; Ricardo Fernández; Mónica Imarai; Tanya Neira; Ximena Lopez; Felipe E. Rodríguez; Antonello Penna; Elias Leiva-Salcedo; Claudio Coddou

    2011-01-01

    The purinergic P2X7 receptor (P2X7R) plays an important role during the immune response, participating in several events such as cytokine release, apoptosis, and necrosis. The bacterial endotoxin lipopolysaccharide (LPS) is one of the strongest stimuli of the immune response, and it has been shown that P2X7R activation can modulate LPS-induced responses. Moreover, a C-terminal binding site for LPS has been proposed. In order to evaluate if LPS can directly modulate the activity of the P2X7R, ...

  2. P2X7 receptor activation amplifies lipopolysaccharide-induced vascular hyporeactivity via interleukin-1? release

    OpenAIRE

    CHIAO, CHIN-WEI; Tostes, Rita C; Webb, R Clinton

    2008-01-01

    Lipopolysaccharide (LPS) stimulates cytoplasmic accumulation of pro-interleukin (IL)-1?. Activation of P2X7 receptors stimulates conversion of pro-IL-1? into mature IL-1?, which is then secreted. Because both LPS (in vivo) and IL-1? (in vitro) decrease vascular reactivity to contractile agents, we hypothesized that: 1) P2X7 receptor activation contributes to LPS-induced vascular hyporeactivity; and 2) IL-1? mediates this change. Thoracic aortas were obtained from 12-week-old male C57BL/6 mice...

  3. Lipopolysaccharide Extracellular Emulsifier Produced by Penicillium citrinum

    OpenAIRE

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

    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...

  4. Lipopolysaccharide Biosynthesis without the Lipids: Recognition Promiscuity of Escherichia coli Heptosyltransferase I

    OpenAIRE

    Czyzyk, Daniel J.; Liu, Cassie; Taylor, Erika A.

    2011-01-01

    Heptosyltransferase I (HepI) is responsible for the transfer of L-glycero-D-manno-heptose to a 3-deoxy-?-D-oct-2-ulopyranosonic acid (Kdo) of the growing core region of lipopolysaccharide (LPS). The catalytic efficiency of HepI with the fully deacylated analogue of Escherichia coli HepI LipidA is 12-fold greater than with the fully acylated substrate, with a kcat/Km of 2.7 × 106 M?1 s?1, compared to a value of 2.2 × 105 M?1 s?1 for the Kdo2-LipidA substrate. Not only is this is the first demo...

  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 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.; Barkholt, Vibeke; Frøkiær, Hanne

    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 pr...

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

    OpenAIRE

    Yuan Yuan, Mei Sun, Ke-Shen Li

    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 m...

  8. A novel Escherichia coli lipid A mutant that produces an antiinflammatory lipopolysaccharide.

    OpenAIRE

    Somerville, J E; Cassiano, L; Bainbridge, B; Cunningham, M. D.; Darveau, R P

    1996-01-01

    A unique screen was used to identify mutations in Escherichia coli lipid A biosynthesis that result in a decreased ability to stimulate E-selectin expression by human endothelial cells. A mutation was identified in the msbB gene of E. coli that resulted in lipopolysaccharide (LPS) that lacks the myristoyl fatty acid moiety of the lipid A. Unlike all previously reported lipid A mutants, the msbB mutant was not conditionally lethal for growth. Viable cells or purified LPS from an msbB mutant ha...

  9. 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

  10. The Klebsiella pneumoniae wabG Gene: Role in Biosynthesis of the Core Lipopolysaccharide and Virulence

    OpenAIRE

    Izquierdo, Luis; Coderch, Núria; Piqué, Nuria; Bedini, Emiliano; Michela Corsaro, Maria; Merino, Susana; Fresno, Sandra; Tomás, Juan M.; Regué, Miguel

    2003-01-01

    To determine the function of the wabG gene in the biosynthesis of the core lipopolysaccharide (LPS) of Klebsiella pneumoniae, we constructed wabG nonpolar mutants. Data obtained from the comparative chemical and structural analysis of LPS samples obtained from the wild type, the mutant strain, and the complemented mutant demonstrated that the wabG gene is involved in attachment to ?-l-glycero-d-manno-heptopyranose II (l,d-HeppII) at the O-3 position of an ?-d-galactopyranosyluronic acid (?-d-...

  11. Dodecaprenyl Phosphate-Galacturonic Acid as a Donor Substrate for Lipopolysaccharide Core Glycosylation in Rhizobium leguminosarum*

    OpenAIRE

    Kanjilal-Kolar, Suparna; Raetz, Christian R H

    2006-01-01

    The lipid A and inner core regions of Rhizobium leguminosarum lipopolysaccharide contain four galacturonic acid (GalA) residues. Two are attached to the outer unit of the 3-deoxy-D-manno-octulosonic acid (Kdo) disaccharide, one to the mannose residue, and one to the 4?-position of lipid A. The enzymes RgtA and RgtB, described in the accompanying article, catalyze GalA transfer to the Kdo residue, whereas RgtC is responsible for modification of the core mannose unit. Heterologous expression of...

  12. Investigations of hippocampal astrocytes in lipopolysaccharide-preconditioned rats in the pilocarpine model of epilepsy

    OpenAIRE

    Bo?ena Pawlikowska-Pawl?ga; Aleksandra Krawczyk; Regina Cybulska; Miros?awa Dmowska; Jadwiga Jaworska-Adamu

    2011-01-01

    The present paper is the first work to determine the effect of lipopolysaccharide (LPS) in the pilocarpine model of epilepsy on the morphology of rat hippocampal astrocytes in vivo. The study involved adult male Wistar rats, which 72 hours prior to administration of pilocarpine hydrochloride (PILO) were intraperitoneally (ip) preconditioned with LPS at a dose of 0.5 mg/kg b.w. The control animals were administered (ip) saline or LPS alone. The astrocytes in the control anim...

  13. Genes of the adaptive immune system are expressed early in zebrafish larval development following lipopolysaccharide stimulation

    Science.gov (United States)

    Li, Fengling; Zhang, Shicui; Wang, Zhiping; Li, Hongyan

    2011-03-01

    Information regarding immunocompetence of the adaptive immune system (AIS) in zebrafish Danio rerio remains limited. Here, we stimulated an immune response in fish embryos, larvae and adults using lipopolysaccharide (LPS) and measured the upregulation of a number of AIS-related genes ( Rag2, AID, TCRAC, IgLC-1, mIg, sIg, IgZ and DAB) 3 and 18 h later. We found that all of the genes evaluated were strongly induced following LPS stimulation, with most of them responding at 8 d post fertilization. This confirms that a functional adaptive immune response is present in D. rerio larvae, and provides a window for further functional analyses.

  14. Enhancement of Natural Resistance to Influenza Virus in Lipopolysaccharide-Responsive and -Nonresponsive Mice by Propionibacterium acnes

    OpenAIRE

    Gangemi, J. David; Hightower, James A.; Jackson, Ronnie A.; Maher, Michael H.; Welsh, Marcia G.; Sigel, M. Michael

    1983-01-01

    Lipopolysaccharide-responsive C3H/HeN mice were rendered resistant to a mouse-adapted strain of influenza (Aichi, H3N2) virus when Propionibacterium acnes was given either intranasally or intraperitoneally several days before virus infection. The time of P. acnes treatment was important since no protection was demonstrated when this agent was given either on the same day as or several days after virus challenge. In contrast, lipopolysaccharide-nonresponsive C3H/HeJ mice were not protected whe...

  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. [A comparative analysis of the ice nucleation activity of pseudomonad cells and lipopolysaccharides].

    Science.gov (United States)

    Zdorovenko, G M; Vereme?chenko, S N; Kipriianova, E A

    2004-01-01

    The paper deals with the study of the ice nucleation activity of the cells, extracellular lipopolysaccharides (ELPSs), lipopolysaccharides (LPSs), and their structural components (lipid A, core oligosaccharide, and O-specific polysaccharide) of Pseudomonas fluorescens, P. syringae, P.fragi, and P. pseudoalcaligenes. The aqueous suspensions of the intact cells of P. syringae IMV 1951 and IMV 185 began to freeze at -1 and -4 degrees C, respectively. This suggests that these cells possess ice nucleation activity. The aqueous cell suspensions of two other strains, P. fluorescens IMV 1433 and IMV 2125, began to freeze at lower temperatures than did distilled water (-9 degrees C), which suggests that the cells of these strains possess antifreeze activity. The ice nucleation activity of the bacterial strains studied did not show any correlation with their taxonomic status. The ice nucleation activity of ELPSs depended little on their concentration (within a concentration range of 0.2-0.4%). In most cases, the ice nucleation activity of ELPSs, LPSs, and their structural components differed from that of the intact cells from which these biopolymers were obtained. This may indicate that the biopolymers under study play a role in ice nucleation, but this role is not crucial. The relationship between the structure of LPSs and their effect on ice nucleation is discussed. PMID:15521177

  17. 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

  18. 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) (.svg) (.html) (.c ... hors Palsson-McDermott EM, O'Neill LA. Publication Immunology . 2004 Oct;113(2):153-62. Pathway - PNG File (.png) ...

  19. The O-specific polysaccharide structure from the lipopolysaccharide of the Gram-negative bacterium Raoultella terrigena.

    Science.gov (United States)

    Leone, Serena; Molinaro, Antonio; Dubery, Ian; Lanzetta, Rosa; Parrilli, Michelangelo

    2007-08-13

    The structure of the repeating unit of the O-specific polysaccharide from the lipopolysaccharide of the enterobacterium Raoultella terrigena was determined by means of chemical and spectroscopical methods and was found to be a linear tetrasaccharide containing a cyclic acetal of pyruvic acid (Pyr) as depicted below.[Carbohydrate structure: see text]. PMID:17509546

  20. MOLECULAR DYNAMICS STUDY OF INTERACTIONS OF POLYMYXIN B3 AND ITS ALA-MUTANTS WITH LIPOPOLYSACCHARIDE

    Directory of Open Access Journals (Sweden)

    Lisnyak Yu. V.

    2015-12-01

    Full Text Available Introduction. Emergence of nosocomial bacterial pathogens (especially Gram-negative bacteria with multiple resistance against almost all available antibiotics is a growing medical problem. No novel drugs targeting multidrug-resistant Gram-negative bacteria have been developed in recent years. In this context, there has been greatly renewed interest to cyclic lipodecapeptides polymyxins. Polymyxins exhibit rapid bactericidal activity, they are specific and highly potent against Gramnegative bacteria, but have potential nephrotoxic side effects. So polymyxins are attractive lead compounds to develop analogues with improved microbiological, pharmacological and toxicological properties. A detailed knowledge of the molecular mechanisms of polymyxin interactions with its cell targets is a prerequisite for the purposeful improvement of its therapeutic properties. The primary cell target of a polymyxin is a lipopolysaccharide (LPS in the outer membrane of Gram-negative bacteria. The binding site of polymyxin on LPS has been supposed to be Kdo2-lipid A fragment. Methods. For all molecular modeling and molecular dynamics simulation experiments the YASARA suite of programs was used. Complex of antimicrobial peptide polymyxin ?3 (PmB3 with Kdo2-lipid A portion of E. coli lipopolysaccharide was constructed by rigid docking with flexible side chains of the peptide. By alanine scanning of polymyxin ?3 bound to LPS followed by simulated annealing minimization of the complexes in explicit water environment, the molecular aspects of PmB3-LPS binding have been studied by 20 ns molecular dynamics simulations at 298 K and pH 7.0. The AMBER03 force field was used with a 1.05 nm force cutoff. To treat long range electrostatic interactions the Particle Mesh Ewald algorithm was used. Results. Ala-mutations of polymyxin’s residues Dab1, Dab3, Dab5, Dab8 and Dab9 in the PmB3-LPS complex caused sustained structural changes resulting in the notable loss in stability of LPS complexes with Ala-mutants of PmB3. The mutations disturbed the characteristic hydrogen-bond network of PmB3-LPS complex. Ala-mutations of Dab1, Dab8 and Dab9 amino acid residues of PmB3 destabilized PmB3- LPS complex to a greater extent: the values of binding energy for these mutants showed increase and largeamplitude irregular fluctuations. Conclusions. Hydrogen bonding of polymyxin B with the lipopolysaccharide is an important factor of the stability of PmB3-LPS complex. Detailed knowledge of the peculiarities of molecular interactions of polymyxins with its primary target on the outer membrane of Gramnegative bacteria is a prerequisite of a purposeful design of novel polymyxin-like lipopeptides.

  1. Anti-lipopolysaccharide toxin therapy for whole body X-irradiation overdose

    International Nuclear Information System (INIS)

    Death in humans from ionising radiation overexposure in the 3-8 Gy (300-800 rad) range is in part due to the toxaemia caused by the entry of gram-negative bacteria and/or their lipopolysaccharide toxin (LPS) into the blood circulation through the walls of partially denuded gut. Anti-LPS hyperimmune equine plasma was evaluated for its ability to lower irradiation-induced lethality. Mice were irradiated with 6.3 Gy (630 rad) and six days later received equine Anti-LPS hyperimmune plasma, control plasma or saline. Mortalities in the three groups were 58%, 92% and 79% (p<0.01) respectively. Thus Anti-LPS may prove useful as an adjunct to conventional therapy in treating radiation sickness. (author)

  2. Immunochemical studies of oligosaccharides obtained from the lipopolysaccharide of Brucella ovis.

    Science.gov (United States)

    Suarez, C E; Pacheco, G A; Vigliocco, A M

    1990-05-01

    Brucella ovis rough lipopolysaccharide (R-LPS) was studied with respect to its heterogeneity, chain length, sugar composition and immunological activity. R-LPS was mildly hydrolysed and oligosaccharides were recovered in the upper phase after partition with chloroform-methanol. Gel-filtration of the upper phase in a column of Bio-Gel P-2 yielded oligosaccharides of 2, 4, 6 and 7 monosaccharide units, 2-keto-deoxy-octulosonic acid (KDO), and monosaccharides. Strong acid hydrolysis followed by paper chromatography showed that the hexa- and heptasaccharides are both composed of glucose, KDO and an unidentified sugar while tetrasaccharide is composed of glucose, mannose and glucosamine. These three oligosaccharides were able to inhibit the LPS-antibody reaction in a solid phase radioimmunoassay, suggesting the oligosaccharides bear antigenic determinants of LPS. PMID:2363245

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

    Directory of Open Access Journals (Sweden)

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

    2012-06-01

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

  4. Importance of Lipopolysaccharide and Cyclic ?-1,2-Glucans in Brucella-Mammalian Infections.

    Science.gov (United States)

    Haag, Andreas F; Myka, Kamila K; Arnold, Markus F F; Caro-Hernández, Paola; Ferguson, Gail P

    2010-01-01

    Brucella species are the causative agents of one of the most prevalent zoonotic diseases: brucellosis. Infections by Brucella species cause major economic losses in agriculture, leading to abortions in infected animals and resulting in a severe, although rarely lethal, debilitating disease in humans. Brucella species persist as intracellular pathogens that manage to effectively evade recognition by the host's immune system. Sugar-modified components in the Brucella cell envelope play an important role in their host interaction. Brucella lipopolysaccharide (LPS), unlike Escherichia coli LPS, does not trigger the host's innate immune system. Brucella produces cyclic ?-1,2-glucans, which are important for targeting them to their replicative niche in the endoplasmic reticulum within the host cell. This paper will focus on the role of LPS and cyclic ?-1,2-glucans in Brucella-mammalian infections and discuss the use of mutants, within the biosynthesis pathway of these cell envelope structures, in vaccine development. PMID:21151694

  5. 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

  6. Analyses of performance of novel sensors with different coatings for detection of Lipopolysaccharide

    KAUST Repository

    Mohd. Syaifudin, A. R.

    2011-10-01

    Interdigital sensors have been widely used for non-destructive applications. New types of planar interdigital sensors have been fabricated with different coating materials to assess the response to Lipopolysaccharide, LPS. All the coatings were selected and optimized to be stable in water, as the measurements take place in water media. Moreover, the coatings have been designed to have available carboxylic or amine functional groups. The use of these functional groups is a widely used technique to specifically binding of biomolecules. The coated sensors were then immobilized with Polymyxin B(PmB) which has the specific binding properties to LPS. This paper will highlight the fabrication process and initial investigations on the sensors\\' performance based on Impedance Spectroscopy. © 2011 IEEE.

  7. 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.

  8. Lipopolysaccharide biosynthesis without the lipids: recognition promiscuity of Escherichia coli heptosyltransferase I.

    Science.gov (United States)

    Czyzyk, Daniel J; Liu, Cassie; Taylor, Erika A

    2011-12-13

    Heptosyltransferase I (HepI) is responsible for the transfer of l-glycero-d-manno-heptose to a 3-deoxy-?-D-oct-2-ulopyranosonic acid (Kdo) of the growing core region of lipopolysaccharide (LPS). The catalytic efficiency of HepI with the fully deacylated analogue of Escherichia coli HepI LipidA is 12-fold greater than with the fully acylated substrate, with a k(cat)/K(m) of 2.7 × 10(6) M(-1) s(-1), compared to a value of 2.2 × 10(5) M(-1) s(-1) for the Kdo(2)-LipidA substrate. Not only is this is the first demonstration that an LPS biosynthetic enzyme is catalytically enhanced by the absence of lipids, this result has significant implications for downstream enzymes that are now thought to utilize deacylated substrates. PMID:22059588

  9. Cytokine release by lipopolysaccharide-stimulated whole blood from patients with typhoid fever.

    Science.gov (United States)

    House, Deborah; Chinh, Nguyen T; Hien, Tran T; Parry, Christopher P; Ly, Nguyen T; Diep, To S; Wain, John; Dunstan, Sarah; White, Nicholas J; Dougan, Gordon; Farrar, Jeremy J

    2002-07-15

    The ex vivo cytokine response to lipopolysaccharide (LPS) of whole blood from patients with typhoid fever was investigated. Tumor necrosis factor (TNF)-alpha release by LPS-stimulated blood was found to be lower during acute typhoid fever than after a course of antimicrobial therapy (P

  10. Responses of equine neutrophils to contagious equine metritis organism and its lipopolysaccharides.

    Science.gov (United States)

    Bertram, T A; Jensen, A E

    1984-06-01

    Morphology and function of equine neutrophils were evaluated after combination with contagious equine metritis organism (CEMO) or 1 of 2 CEMO lipopolysaccharides (LPS). The 2 LPS (LPS-a; LPS-p) isolated from the CEMO contained 14- and 16-carbon fatty acids, ketodeoxyoctanate, hexose, and heptose, but were morphologically distinct. Neutrophils exposed to LPS had fewer granules, whereas those exposed to CEMO had more granules than did the controls (phosphate-buffered saline solution). Neutrophil iodination was significantly increased with 10 and 25 micrograms of LPS-a, but not significantly altered by LPS-p or CEMO. Staphylococcus aureus ingestion was not influenced by CEMO and was mildly decreased by LPS-a. These results indicate that CEMO may have at least 2 functionally and morphologically distinct, but chemically similar, LPS and that 1 of these LPS (LPS-a) may enhance neutrophil killing by stimulating neutrophil iodinating mechanisms. PMID:6742570

  11. Lipopolysaccharide-induced experimental immune activation does not impair memory functions in humans.

    Science.gov (United States)

    Grigoleit, Jan-Sebastian; Oberbeck, J Reiner; Lichte, Philipp; Kobbe, Philipp; Wolf, Oliver T; Montag, Thomas; del Rey, Adriana; Gizewski, Elke R; Engler, Harald; Schedlowski, Manfred

    2010-11-01

    Systemic immune activation occurring together with release of peripheral cytokines can affect behavior and the functioning of the central nervous system (CNS). However, it remains unknown whether and to what extent cognitive functions like memory and attention are affected during transient immune activation. We employed a human endotoxemia model and standardized neuropsychological tests to assess the cognitive effects of an experimental inflammation in two groups of 12 healthy young men before and after intravenous injection of lipopolysaccharide (LPS, Escherichia coli, 0.4 ng/kg) or physiological saline. Endotoxin administration caused a profound transient physiological response with elevations in body temperature, number of circulating neutrophils, and increases in plasma cytokine levels [interleukin (IL)-6, IL-10, tumor necrosis factor (TNF)-?], and concentrations of norepinephrine, ACTH and cortisol. However, these changes in immune and neuroendocrine parameters were not associated with alterations of memory performance, selective attention or executive functions. PMID:20875866

  12. Structural and thermodynamic analyses of the interaction between melittin and lipopolysaccharide.

    Science.gov (United States)

    Bhunia, Anirban; Domadia, Prerna N; Bhattacharjya, Surajit

    2007-12-01

    Lipopolysaccharide (LPS), the major constituent of the outer membrane of Gram-negative bacteria, is the very first site of interactions with the antimicrobial peptides. In this work, we have determined a solution conformation of melittin, a well-known membrane active amphiphilic peptide from honey bee venom, by transferred nuclear Overhauser effect (Tr-NOE) spectroscopy in its bound state with lipopolysaccharide. The LPS bound conformation of melittin is characterized by a helical structure restricted only to the C-terminus region (residues A15-R24) of the molecule. Saturation transfer difference (STD) NMR studies reveal that several C-terminal residues of melittin including Trp19 are in close proximity with LPS. Isothermal titration calorimetry (ITC) data demonstrates that melittin binding to LPS or lipid A is an endothermic process. The interaction between melittin and lipid A is further characterized by an equilibrium association constant (Ka) of 2.85 x 10(6) M(-1) and a stoichiometry of 0.80, melittin/lipid A. The estimated free energy of binding (delta G0), -8.8 kcal mol(-1), obtained from ITC experiments correlates well with a partial helical structure of melittin in complex with LPS. Moreover, a synthetic peptide fragment, residues L13-Q26 or mel-C, derived from the C-terminus of melittin has been found to contain comparable outer membrane permeabilizing activity against Escherichia coli cells. Intrinsic tryptophan fluorescence experiments of melittin and mel-C demonstrate very similar emission maxima and quenching in presence of LPS micelles. The Red Edge Excitation Shift (REES) studies of tryptophan residue indicate that both peptides are located in very similar environment in complex with LPS. Collectively, these results suggest that a helical conformation of melittin, at its C-terminus, could be an important element in recognition of LPS in the outer membrane. PMID:17854761

  13. 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.

  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. Shrimp (Penaeus monodon) anti-lipopolysaccharide factor reduces the lethality of Pseudomonas aeruginosa sepsis in mice.

    Science.gov (United States)

    Pan, Chia-Yu; Chao, Tsung-Tai; Chen, Jian-Chyi; Chen, Jyh-Yih; Liu, Wei-Chen; Lin, Cheng-Hui; Kuo, Ching-Ming

    2007-05-01

    We investigated the efficacy of amino acids 55-76 of the synthetic shrimp anti-lipopolysaccharide factor peptide (SALF(55-76) cyclic peptide), the C-terminal part of the shrimp anti-lipopolysaccharide factor. This study was conducted to elucidate the effects of the antiseptic action of this peptide. The SALF(55-76) cyclic peptide was tested against bacterial clinical isolates and showed broad-spectrum antimicrobial activity. Transmission electron microscopic (TEM) examination of SALF(55-76) cyclic peptide-treated Pseudomonas aeruginosa showed that severe swelling preceded cell death and breakage of the outer membrane; the intracellular inclusion was found to have effluxed extracellularly. When mice were treated with the SALF(55-76) cyclic peptide before bacterial challenge with P. aeruginosa, the peptide highly protected mice against death by sepsis. The P. aeruginosa recovered from SALF(55-76) cyclic peptide-treated mice after 4 h exhibited reduced bacterial growth similar to that recovered from vancomycin-treated mice. In addition, the syntheses of inflammatory cytokines, such as interleukin (IL)-2, IL-4, IL-10, IL-12, IL-13, interferon-gamma, and tumor necrosis factor [TNF]-alpha, were significantly upregulated 4 h after SALF(55-76) cyclic peptide treatment except for IL-4 in the liver. The expressions of Toll-like receptor 4 (Tlr4), Irf3, myd88, and Tram, were considerably elevated, but only Tlr4 existed in the spleen 4 h after SALF(55-76) cyclic peptide treatment. The prophylactic administration of SALF(55-76) cyclic peptide was begun the TNF-alpha response in comparison to untreated mice by an ELISA analysis. Due to its multifunctional properties, the SALF(55-76) cyclic peptide may become an important prophylaxis against and therapy for bacterial infectious diseases, as well as for septic shock. PMID:17386416

  16. Antimicrobial Activity of Diterpenes from Viguiera arenaria against Endodontic Bacteria

    OpenAIRE

    Martins, Carlos H. G.; Erika Borges dos Reis; Maria Gorete M. Souza; Brenda P. F. A. Gomes; Da Costa, Fernando B.; Vladimir C. G. Heleno; RODRIGO C. S. VENEZIANI; Niege A. J. C. Furtado; Ambrósio, Sérgio R.; Tatiane C. de Carvalho; Marília R. Simão

    2011-01-01

    Six pimarane-type diterpenes isolated from Viguiera arenaria Baker and two semi-synthetic derivatives were evaluated in vitro against a panel of representative microorganisms responsible for dental root canal infections. The microdilution method was used for the determination of the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against Porphyromonas gingivalis, Prevotella nigrescens, Prevotella intermedia, Prevotella buccae, Fusobacterium nucleatum, Bacte...

  17. Mouthrinses: a comparative microbiological study

    OpenAIRE

    Gautier, G.; Noguer Castellvi, Miguel; Costa, N.; Canela, J.; Viñas, Miquel

    1999-01-01

    This study was performed in order to evaluate the efficacy of different mouthrinses whose use is extended in Spain. Six different antiseptic mouthrinses were studied by means of determination of Minimal Inhibitory Concentration (MIC) values against Klebsiella pneumoniae, Serratia marcescens, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella typhimurium, Bacillus subtilis, Streptococcus mutans, Prevotella intermedia, Porphyromonas gingivalis and Actinobacillus actinom...

  18. 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)...

  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. Nitric oxide production by hemocytes of larva and pharate prepupa of Galleria mellonella in response to bacterial lipopolysaccharide: cytoprotective or cytotoxic?.

    Czech Academy of Sciences Publication Activity Database

    Krishnan, Natraj; Hyršl, P.; Šimek, V.

    2006-01-01

    Ro?. 142, 1-2 (2006), s. 103-110. ISSN 1532-0456 Institutional research plan: CEZ:AV0Z50070508 Keywords : nitric oxide * hemocytes * lipopolysaccharide Subject RIV: ED - Physiology Impact factor: 1.991, year: 2006

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

    Lifescience Database Archive (English)

    Full Text Available 1373512 Function of lipopolysaccharide (LPS)-binding protein (LBP) and CD14, thereceptor for LPS ... BP complexes: a short review. Authors Schumann RR. Publication ... Res Immunol. 1992 Jan;143(1):11-5. Pathway - PNG F ...

  2. Black Tea Extract and Its Theaflavin Derivatives Inhibit the Growth of Periodontopathogens and Modulate Interleukin-8 and ?-Defensin Secretion in Oral Epithelial Cells

    Science.gov (United States)

    Lombardo Bedran, Telma Blanca; Morin, Marie-Pierre; Palomari Spolidorio, Denise; Grenier, Daniel

    2015-01-01

    Over the years, several studies have brought evidence suggesting that tea polyphenols, mostly from green tea, may have oral health benefits. Since few data are available concerning the beneficial properties of black tea and its theaflavin derivatives against periodontal disease, the objective of this study was to investigate their antibacterial activity as well as their ability to modulate interleukin-8 and human ?-defensin (hBD) secretion in oral epithelial cells. Among the periodontopathogenic bacteria tested, Porphyromonas gingivalis was found to be highly susceptible to the black tea extract and theaflavins. Moreover, our data indicated that the black tea extract, theaflavin and theaflavin-3,3’-digallate can potentiate the antibacterial effect of metronidazole and tetracycline against P. gingivalis. Using lipopolysaccharide-stimulated oral epithelial cells, the black tea extract (100 ?g/ml), as well as theaflavin and theaflavin-3,3’-digallate (50 ?g/ml) reduced interleukin-8 (IL-8) secretion by 85%, 79%, and 86%, respectively, thus suggesting an anti-inflammatory property. The ability of the black tea extract and its theaflavin derivatives to induce the secretion of the antimicrobial peptides hBD-1, hBD-2 and hBD-4 by oral epithelial cells was then evaluated. Our results showed that the black tea extract as well as theaflavin-3,3’-digallate were able to increase the secretion of the three hBDs. In conclusion, the ability of a black tea extract and theaflavins to exert antibacterial activity against major periodontopathogens, to attenuate the secretion of IL-8, and to induce hBD secretion in oral epithelial cells suggest that these components may have a beneficial effect against periodontal disease. PMID:26581041

  3. Lipopolysaccharide-induced depressive-like behavior is mediated by indoleamine 2,3-dioxygenase activation in mice

    OpenAIRE

    O’Connor, J.C.; Lawson, M.A.; André, C.; Moreau, M.; Lestage, J.; Castanon, N.; Kelley, K. W.; Dantzer, R.

    2008-01-01

    Although elevated activity of the tryptophan-degrading enzyme indoleamine 2,3-dioxygenase(IDO) has been proposed to mediate comorbid depression in inflammatory disorders, its causative role has never been tested. We report that peripheral administration of lipopolysaccharide (LPS) activates IDO and culminates in a distinct depressive-like behavioral syndrome, measured by increased duration of immobility in both the forced swim and tail suspension tests. Blockade of IDO activation either indir...

  4. 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...

  5. Selective activation and functional significance of p38? mitogen-activated protein kinase in lipopolysaccharide-stimulated neutrophils

    OpenAIRE

    Nick, Jerry A.; Avdi, Natalie J.; Young, Scott K.; Lehman, Lisa A.; McDonald, Patrick P; Frasch, S. Courtney; Billstrom, Marcella A.; Henson, Peter M; JOHNSON, GARY L.; Worthen, G Scott

    1999-01-01

    Activation of leukocytes by proinflammatory stimuli selectively initiates intracellular signal transduction via sequential phosphorylation of kinases. Lipopolysaccharide (LPS) stimulation of human neutrophils is known to result in activation of p38 mitogen-activated protein kinase (MAPk); however, the upstream activator(s) of p38 MAPk is unknown, and consequences of p38 MAPk activation remain largely undefined. We investigated the MAPk kinase (MKK) that activates p38 MAPk in response to LPS, ...

  6. Coxiella burnetii lipopolysaccharide blocks p38?-MAPK activation through the disruption of TLR-2 and TLR-4 association

    OpenAIRE

    Conti, Filippo; Boucherit, Nicolas; Baldassarre, Veronica; Trouplin, Virginie; Toman, Rudolf; Mottola, Giovanna; Mege, Jean-Louis; Ghigo, Eric

    2015-01-01

    To survive in macrophages, Coxiella burnetii hijacks the activation pathway of macrophages. Recently, we have demonstrated that C. burnetii, via its lipopolysaccharide (LPS), avoids the activation of p38?-MAPK through an antagonistic engagement of Toll-like receptor (TLR)-4. We investigated the fine-tuned mechanism leading to the absence of activation of the p38?-MAPK despite TLR-4 engagement. In macrophages challenged with LPS from the avirulent variants of C. burnetii, TLR-4 and TLR-2 co-im...

  7. Inhibitory Activities of New Series of 4, 5-diaryl Thiadiazoles Derivatives on Lipopolysaccharide-induced Cox-2 Expression

    OpenAIRE

    Seyed Nasser Ostad; Mohsen Amini; Zahra Haghipour; Leila Karimi; Latifeh Navidpour

    2005-01-01

    Recent studies have suggested cyclooxygenase-2 (COX-2) expression as a mechanism involved in carcinogenesis. It has also been suggested that changes in COX-2 expression level can be considered as a possible therapeutic target in tumors. Therefore, it was decided to synthesize a new series of 4, 5-diaryl thiadiazoles (compounds 1-5) as COX-2 inhibitors and evaluate their inhibitory activity on COX-2 expression. The COX-2 expression was induced by lipopolysaccharide (LPS) in bovine aortic endot...

  8. Serum Amyloid P Component Bound to Gram-Negative Bacteria Prevents Lipopolysaccharide-Mediated Classical Pathway Complement Activation

    OpenAIRE

    de Haas, Carla J.C.; van Leeuwen, Ester M.M.; van Bommel, Toon; Verhoef, Jan; van Kessel, Kok P.M.; van Strijp, Jos A.G.

    2000-01-01

    Although serum amyloid P component (SAP) is known to bind many ligands, its biological function is not yet clear. Recently, it was demonstrated that SAP binds to lipopolysaccharide (LPS). In the present study, SAP was shown to bind to gram-negative bacteria expressing short types of LPS or lipo-oligosaccharide (LOS), such as Salmonella enterica serovar Copenhagen Re and Escherichia coli J5, and also to clinical isolates of Haemophilus influenzae. It was hypothesized that SAP binds to the bact...

  9. Clinico-pathological Changes Associated with Brucella melitensis Infection and its Bacterial Lipopolysaccharides (LPS) in Male Mice

    OpenAIRE

    Faez Firdaus Jesse Abdullah; Norasiah Binti Nik; Mohd Zamri Saad; Abd. Wahid Haron; Abdul Rahman Omar; Jasni Sabri; Lawan Adamu; Abdinasir Yusuf Osman; Abdul Aziz Saharee

    2013-01-01

    Brucella melitensis (B. melitensis) is gram negative, aerobic bacteria that cause Brucellosis in humans’ sheep and goats. Brucellosis causes abortion in wild and domestic animals resulting in enormous financial losses. Therefore, the purpose of this study was to evaluate the clinico-pathological changes associated with Brucella melitensis infection and its bacterial Lipopolysaccharides (LPS) in male mice. Three groups of 24 Balb/c male mice consisting of 8 mice in each group were used as an a...

  10. Identification and Characterization of the Brucella abortus Phosphoglucomutase Gene: Role of Lipopolysaccharide in Virulence and Intracellular Multiplication

    OpenAIRE

    Ugalde, Juan E.; Czibener, Cecilia; Feldman, Mario F; Ugalde, Rodolfo A

    2000-01-01

    Smooth lipopolysaccharide (LPS) of Brucella abortus has been reported to be an important virulence factor, although its precise role in pathogenesis is not yet clear. While the protective properties of LPS against complement are well accepted, there is still some controversy about the capacity of rough mutants to replicate intracellularly. The B. abortus phosphoglucomutase gene (pgm) was cloned, sequenced, and disrupted. The gene has a high index of identity to Agrobacterium tumefaciens pgm b...

  11. Comparison of Protective Efficacy of Subcutaneous versus Intranasal Immunization of Mice with a Brucella melitensis Lipopolysaccharide Subunit Vaccine

    OpenAIRE

    APURBA K. BHATTACHARJEE; Izadjoo, Mina J.; Wendell D. Zollinger; Nikolich, Mikeljon P.; Hoover, David L.

    2006-01-01

    Groups of mice were immunized either subcutaneously or intranasally with purified Brucella melitensis lipopolysaccharide (LPS) or with LPS as a noncovalent complex with Neisseria meningitidis group B outer membrane protein (LPS-GBOMP). Control mice were inoculated with sterile saline. Two doses of vaccine were given 4 weeks apart. Mice were challenged intranasally with virulent B. melitensis strain 16M 4 weeks after the second dose of vaccine. Sera, spleens, lungs, and livers of mice were har...

  12. Pantoea agglomerans lipopolysaccharide maintains bone density in premenopausal women: a randomized, double-blind, placebo-controlled trial

    OpenAIRE

    Nakata, Kazue; Nakata, Yoko; Inagawa, Hiroyuki; Nakamoto, Takeru; Yoshimura, Hiroshi; Soma, Gen-ichiro

    2014-01-01

    Lipopolysaccharide fromPantoea agglomerans (LPSp) facilitates Ca and P turnover in chicken calvaria and femurs. This study investigated osteoporosis prevention by the oral administration of LPSp in mice and in double-blind clinical tests. Using ovariectomized (OVX) osteoporosis mice model, we investigated the effects of LPSp on the bone density and Ca concentration after ingesting LPSp-containing water for 4 weeks. Oral administration of LPSp tended to suppress the decline in the bone density...

  13. Inhibition of mitochondrial permeability transition by cyclosporin A prevents pyrazole plus lipopolysaccharide-induced liver injury in mice

    OpenAIRE

    Zhuge, Jian; Cederbaum, Arthur I

    2008-01-01

    Previous results showed that pyrazole potentiates lipopolysaccharide (LPS)-induced liver injury in mice. Mechanisms involved overexpression of cytochrome P450 2E1 (CYP2E1), oxidative stress, activation of c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MAPK). The current study was carried out to test the hypothesis that the mitochondria permeability transition (MPT) plays a role in this pyrazole plus LPS toxicity. Mice were injected intraperitoneally with pyrazole for ...

  14. Cytokine and acute phase protein gene expression in liver biopsies from dairy cows with a lipopolysaccharide - induced mastitis

    DEFF Research Database (Denmark)

    Vels, J; Røntved, Christine M.; Bjerring, Martin; Ingvartsen, Klaus Lønne

    2009-01-01

    A minimally invasive liver biopsy technique was tested for its applicability to study the hepatic acute phase response (APR) in dairy cows with Escherichia coli lipopolysaccharide (LPS)-induced mastitis. The hepatic mRNA expression profiles of the inflammatory cytokines, tumor necrosis factor (TNF- ), IL-1?, IL-6, and IL-10, and the acute phase proteins serum amyloid A isoform 3 (SAA3), haptoglobin (Hp), and 1-acid glycoprotein (AGP) were determined by real-time reverse transcription-PCR. Fourte...

  15. Induction of Murine Gamma Interferon Production by Lipopolysaccharide and Interleukin-2 in Propionibacterium acnes-Induced Peritoneal Exudate Cells

    OpenAIRE

    1987-01-01

    Lipopolysaccharide (LPS) induces high levels of gamma interferon (IFN-gamma) in the circulation of mice pretreated with heat-killed Propionibacterium acnes. The following results were obtained in the present study. LPS, as well as interleukin-2 (IL-2), was also able to induce IFN-gamma in vitro in peritoneal exudate cells (PEC) from such mice. Splenocytes and lymph node cells from these mice or resident peritoneal cells from control mice produced trace or undetectable amount of IFN-gamma upon...

  16. Mitogenic Response of Murine B Lymphocytes to Salmonella typhimurium Lipopolysaccharide Requires Protein Kinase C-Dependent Late Tyrosine Phosphorylations

    OpenAIRE

    Mey, Anne; Revillard, Jean-Pierre

    1998-01-01

    Unlike the cross-linking of membrane immunoglobulins, the activation of B cells by lipopolysaccharide (LPS) does not involve the phosphoinositol turnover and the initial activation of tyrosine kinases. However, LPS-induced B-cell proliferation was inhibited by the tyrosine kinase inhibitors genistein and herbimycin A even when added 48 h after the beginning of the culture. Tyrosyl-phosphorylated proteins were detected by Western blotting after 24 h of culture with LPS, reaching a maximum conc...

  17. Cannabinoid Receptor CB2 Is Involved in Tetrahydrocannabinol-Induced Anti-Inflammation against Lipopolysaccharide in MG-63 Cells

    OpenAIRE

    Yang, Lei; Li, Fei-Fei; Han, Yu-chen; Jia, Bin; Ding, Yin

    2015-01-01

    Cannabinoid ?9-tetrahydrocannabinol (THC) is effective in treating osteoarthritis (OA), and the mechanism, however, is still elusive. Activation of cannabinoid receptor CB2 reduces inflammation; whether the activation CB2 is involved in THC-induced therapeutic action for OA is still unknown. Cofilin-1 is a cytoskeleton protein, participating in the inflammation of OA. In this study, MG-63 cells, an osteosarcoma cell-line, were exposed to lipopolysaccharide (LPS) to mimic the inflammation of O...

  18. Lipopolysaccharide-Based Enzyme-Linked Immunosorbent Assay for Experimental Use in Detection of Antibodies to Lawsonia intracellularis in Pigs

    OpenAIRE

    Kroll, J. J.; Eichmeyer, M. A.; Schaeffer, M L; McOrist, S; Harris, D L; Roof, M B

    2005-01-01

    An enzyme-linked immunosorbent assay (ELISA) for Lawsonia intracellularis was developed and compared with a whole-cell antigen-based immunofluorescence antibody test (IFAT). The antigen-containing lipopolysaccharide (LPS) was derived from Percoll gradient purified cultures of L. intracellularis by using a modification of the Westphal hot phenol procedure. The antigen was bound directly to polystyrene 96-well microtiter plates, and the assay was performed in an indirect ELISA format. Specifici...

  19. Occurrence of 2-keto-3-deoxy-D-manno-octonic acid in lipopolysaccharides isolated from Vibrio parahaemolyticus.

    OpenAIRE

    Han, T J; Chai, T. J.

    1991-01-01

    The occurrence of 2-keto-3-deoxy-D-manno-octonic acid (KDO) in lipopolysaccharides (LPS) of Vibrio parahaemolyticus was demonstrated for the first time by gas chromatography-mass spectrometry after dephosphorylation, reduction, and methylation. KDO was virtually completely phosphorylated, since no KDO was detected by either gas chromatography or thiobarbituric acid assay before dephosphorylation. The level of KDO in all six strains of V. parahaemolyticus investigated ranged from 0.37 to 0.69%...

  20. Morphine preconditioning reduces lipopolysaccharide and interferon-?-induced mouse microglial cell injury via ?1 opioid receptor activation

    OpenAIRE

    Gwak, M-S; Li, L.(Institute of High Energy Physics, Chinese Academy of Sciences, Beijing, China; Department of Modern Physics, University of Science and Technology of China, Hefei, Anhui, China; Department of Physics, Nanjing University, Nanjing, Jiangsu, China; School of Physics, Shandong University, Jinan, Shandong, China; Physics Department, Shanghai Jiao Tong University, Shanghai, China); Zuo, Z

    2010-01-01

    Microglial cells play an important role in the inflammatory response of a broad range of brain diseases including stroke, brain infection and neurodegenerative diseases. However, there is very little information regarding how to protect microglial cells. Here, we showed that incubation of the C8-B4 mouse microglial cells with lipopolysaccharide (LPS) plus interferon-? (IFN?) induced cytotoxicity as assessed by the amount of lactate dehydrogenase (LDH) released from the cells. Preconditioning ...

  1. Pyocin-resistant lipopolysaccharide mutans of Neisseria gonorrhoeae: alterations in sensitivity to normal human serum and polymyxin B.

    OpenAIRE

    Guymon, L F; Esser, M; Shafer, W. M.

    1982-01-01

    Pyocins from Pseudomonas aeruginosa were used to select several lipopolysaccharide (LPS) mutants of Neisseria gonorrhoeae strain FA19. Three classes of LPS mutans were found in the initial group selected for study. The LPS of one class lacked galactose. That of a second group lacked the typical heptose found in the parental LPS, was reduced in glucose, galactose, and N-acetylglucosamine content, appeared to contain a new unidentified sugar component, and consisted of two species of LPS separa...

  2. Synergistic anti-inflammatory effects of Nobiletin and Sulforaphane in lipopolysaccharide-stimulated RAW 264.7 cells

    OpenAIRE

    Guo, Shanshan; Qiu, Peiju; Xu, Guang; Wu, Xian.; DONG, PING; Yang, Guanpin; Zheng, Jinkai; McClements, David Julian; Xiao, Hang

    2012-01-01

    Inflammation plays important roles in initiation and progress of many diseases including cancers in multiple organ sites. Herein, we investigated the anti-inflammatory effects of two dietary compounds, nobiletin (NBN) and sulforaphane (SFN) in combination. Non-cytotoxic concentrations of NBN, SFN, and their combinations were studied in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophage cells. The results showed that combined NBN and SFN treatments produced much stronger inhibitory effec...

  3. The Internalization Time Course of a Given Lipopolysaccharide Chemotype Does Not Correspond to Its Activation Kinetics in Monocytes

    OpenAIRE

    Lentschat, A.; El-Samalouti, V. T.; Schletter, J.; Kusumoto, S; Brade, L.; Rietschel, E. T.; Gerdes, J.; Ernst, M.; Flad, H.-D.; Ulmer, A J

    1999-01-01

    The prerequisites for the initiation of pathophysiological effects of endotoxin (lipopolysaccharide [LPS]) include binding to and possibly internalization by target cells. Monocytes/macrophages are prominent target cells which are activated by LPS to release various pro- and anti-inflammatory mediators. The aim of the present study was to establish a new method to determine the binding and internalization rate of different LPS chemotypes by human monocytes and to correlate these phenomena wit...

  4. Lack of In Vitro and In Vivo Recognition of Francisella tularensis Subspecies Lipopolysaccharide by Toll-Like Receptors?

    OpenAIRE

    Hajjar, Adeline M; Harvey, Megan D.; Shaffer, Scott A.; Goodlett, David R; Sjöstedt, Anders; Edebro, Helen; Forsman, Mats; Byström, Mona; Pelletier, Mark; Wilson, Christopher B.; Miller, Samuel I.; Skerrett, Shawn J.; Ernst, Robert K

    2006-01-01

    Francisella tularensis is an intracellular gram-negative bacterium that is highly infectious and potentially lethal. Several subspecies exist of varying pathogenicity. Infection by only a few organisms is sufficient to cause disease depending on the model system. Lipopolysaccharide (LPS) of gram-negative bacteria is generally recognized by Toll-like receptor 4 (TLR4)/MD-2 and induces a strong proinflammatory response. Examination of human clinical F. tularensis isolates revealed that human vi...

  5. Role of YadA, Ail, and Lipopolysaccharide in Serum Resistance of Yersinia enterocolitica Serotype O:3

    OpenAIRE

    Biedzka-Sarek, Marta; Venho, Reija; Skurnik, Mikael

    2005-01-01

    Complement attack is a host strategy leading to elimination of pathogens. Yersinia enterocolitica expresses several potential complement resistance factors: the outer membrane proteins YadA and Ail as well as lipopolysaccharide (LPS). To study the contribution of these factors to the survival of Y. enterocolitica serotype O:3 in nonimmune human serum, we constructed 23 mutant strains of Y. enterocolitica O:3 expressing different combinations of YadA, Ail, LPS O antigen, and LPS outer core. Su...

  6. Lipopolysaccharide as an Antigen Target for the Formulation of a Universal Vaccine against Escherichia coli O111 Strains ?

    OpenAIRE

    Santos, Maurílio F.; New, Roger R. C.; Andrade, Gabrielle R.; Ozaki, Christiane Y; Sant'Anna, Osvaldo A.; Mendonça-Previato, Lucia; Trabulsi, Luis R.; Domingos, Marta O.

    2010-01-01

    A promising approach to developing a vaccine against O111 strains of diarrheagenic Escherichia coli that exhibit different mechanisms of virulence is to target either the core or the polysaccharide chain (O antigen) of their lipopolysaccharide (LPS). However, due to structural variations found in both these LPS components, to use them as antigen targets for vaccination, it is necessary to formulate a vaccine able to induce a humoral immune response that can recognize all different variants fo...

  7. Lipopolysaccharide differentially decreases plasma acyl and desacyl ghrelin levels in rats: potential role of the circulating ghrelin acylating enzyme GOAT

    OpenAIRE

    Stengel, Andreas; Goebel, Miriam; Wang, Lixin; Reeve, Joseph R.; TACHÉ, YVETTE; Lambrecht, Nils W.G.

    2010-01-01

    Bacterial lipopolysaccharide (LPS) in rodents is an established model for studying innate immune responses to gram-negative bacteria and mimicking symptoms of infections including reduced food intake associated with decreased circulating total ghrelin levels. The ghrelin-acylating enzyme, ghrelin-O-acyltransferase (GOAT) involved in the formation of acyl ghrelin (AG) was recently identified. We investigated changes in circulating AG, desacyl ghrelin (DG) and GOAT induced by intraperitoneal LP...

  8. Identification of lipopolysaccharide-binding peptide regions within HMGB1 and their effects on subclinical endotoxemia in a mouse model

    OpenAIRE

    Youn, Ju Ho; Kwak, Man Sup; Wu, Jie; Kim, Eun Sook; Ji, Yeounjung; Min, Hyun Jin; Yoo, Ji-Ho; CHOI, JI EUN; Cho, Hyun-soo; Shin, Jeon-Soo

    2011-01-01

    Lipopolysaccharide (LPS) triggers deleterious systemic inflammatory responses when released into the circulation. LPS-binding protein (LBP) in the serum plays an important role in modifying LPS toxicity by facilitating its interaction with LPS signaling receptors, which are expressed on the surface of LPS-responsive cells. We have previously demonstrated that high mobility group box 1 (HMGB1) can bind to and transfer LPS, consequently increasing LPS-induced TNF-? production in human periphera...

  9. Prunus yedoensis Bark Inhibits Lipopolysaccharide-Induced Inflammatory Cytokine Synthesis by I?B? Degradation and MAPK Activation in Macrophages

    OpenAIRE

    Yun, Jeong-Moon; Im, Sung-Bin; Roh, Mahn-Kwang; Park, Sung-Hyun; Kwon, Han-Al; Lee, Ju-Yeong; Choi, Ho-Young; HAM, IN-HYE; Kim, Yoon Bum; Lee, Jeoung-Min; Kim, Dae-Ok; Park, Kye Won; Kang, Hee

    2014-01-01

    The bark of Prunus yedoensis is used in antitussive medicines and in oral herbal formulations for inflammatory skin disorders. In the present study, we explored whether P. yedoensis bark extract (PYE) and its solvent partitioned fractions could modulate lipopolysaccharide (LPS)-induced tumor necrosis factor (TNF)-? and interleukin (IL)-6 in vivo and in vitro. In addition, we examined the effect of PYE extract and its fractions on LPS-induced NF-?B and mitogen-activated protein kinase (MAPK) s...

  10. 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

  11. 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.

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

    Science.gov (United States)

    Ma, Zhiwei; Dagnaes-Hansen, Frederik; Løvschall, Henrik; Song, Wen; Nielsen, Gitte K; Yang, Chuanxu; Wang, Qintao; Kjems, Jørgen; Gao, Shan

    2015-08-01

    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. PMID:25258036

  13. Production of interleukin-1 receptor antagonist isoforms by microglia in mixed rat glial cells stimulated by lipopolysaccharide.

    Science.gov (United States)

    Pousset, F; Palin, K; Verrier, D; Bristow, A; Dantzer, R; Parnet, P; Lestage, J

    2000-12-01

    Although the natural interleukin-1 receptor antagonist (IL-1Ra) has been shown to be produced by microglial cells in response to immune stimuli, nothing was known about the ability of these cells in primary culture to produce the different isoforms of IL-1Ra. Using RT-PCR, we first confirmed that mixed glial cell cultures from newborn rats respond to the cytokine inducer, lipopolysaccharide, by synthesizing IL-1Ra mRNA. Using double immunostaining, we showed that IL-1Ra was detected in microglia but not in astrocytes. Using Western blotting, we finally demonstrated that the IL-1Ra1 isoform was secreted in the supernatant of mixed glial cell cultures, and its production increased in response to lipopolysaccharide. The three different IL-1Ra isoforms were constitutively expressed in cell lysates and their levels increased after lipopolysaccharide treatment, except for IL-1Ra3. These results point to the ability of microglial cells in primary culture to produce the different isoforms of IL-1Ra. PMID:11125314

  14. Reconocimiento de antígenos lipopolisacarídicos de Pseudomonas aeruginosa por sueros humanos / Recognises of antigen lipopolysaccharide from Pseudomonas aeruginosa by human sera

    Scientific Electronic Library Online (English)

    Jonatán, Hernández; Jacquelin, Alfonso; Juan F., Almenares; Aniel, Moya; Jose L., Pérez; Sara C., Esnard; Armando, Cádiz; Gustavo, Sierra.

    2004-04-01

    Full Text Available En este trabajo se procedió al reconocimiento de lipopolisacáridos de diferentes serotipos de Pseusodomas aeruginosa por sueros humanos. Los lipopolisacáridos fueron aislados por el método Wespthal y Jann. Se utilizaron un total de 104 sueros de pacientes infectados y donantes voluntarios, provenien [...] tes de instalaciones hospitalarias y del Banco de Sangre, para el reconocimiento de antígenos lipopolisacarídicos, mediante el ensayo de Dott Blot. Los resultados mostraron que los serotipos de mayor reconocimiento, por los sueros testados, fueron el O11, O13, O16 y el O15, existiendo diferencias significativas entre ellos (p Abstract in english ABSTRACT This work presents the recognition of different serotypes of Pseudomonas aeruginosa lipopolysaccharide by human sera. The lipopolysaccharides were purified by the methods of Wespthal and Jann. On the other hand, a total of 104 serums, of infected patients and voluntary donors, coming from d [...] ifferent hospitals and from the Blood Bank of the Marianao municipality were used for the recognition of the antigenic lipopolysaccharides, using of dot blot. Our results showed that the serotypes most frequently recognized by the tested serums, were O11, O13, O16 and O15, with significant differences among them (p

  15. 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.

  16. 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)

  17. 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.

  18. 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.

  19. Deoxynivalenol-induced cytotoxicity, cytokines and related genes in unstimulated or lipopolysaccharide stimulated primary porcine macrophages.

    Science.gov (United States)

    Döll, Susanne; Schrickx, Jan A; Dänicke, Sven; Fink-Gremmels, Johanna

    2009-01-30

    The cytotoxicity of deoxynivalenol (DON) as well as the induction of cytokines and related genes was investigated in porcine pulmonary alveolar macrophages (PAM) in absence or presence of lipopolysaccharides (LPS). IC(20) values were 1.1, 0.4 and 1.0microM DON in the MTT, neutral red and alamar blue assay, respectively, and did not differ significantly in the presence of LPS. The mRNA expression of tumour necrosis factor (TNF)-alpha peaked at 3h, whereas LPS and DON showed synergistic effects resulting in an approximately 20-fold increase at 500nM DON as compared to untreated controls. The supernatant concentrations of TNF-alpha showed similar synergistic effects. The expression of interleukin (IL)-1beta was significantly induced by DON (except for 12h) and LPS. An induction of the mRNA expression of IL-6 by DON was evident only at 3h, whereas the supernatant concentrations of LPS stimulated PAM incubated with 500nM DON were significantly decreased at most time points. Cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) expression did not seem to contribute to the effects of DON in porcine macrophages. The results of the present investigation suggest a contribution of cytokines, especially TNF-alpha and IL-1beta, induced by DON in porcine macrophages to the effects observed in vivo. PMID:19027837

  20. 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

  1. 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

  2. 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

  3. 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.

  4. 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.

  5. 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.

  6. Rat microglial cells secrete predominantly the precursor of interleukin-1beta in response to lipopolysaccharide.

    Science.gov (United States)

    Chauvet, N; Palin, K; Verrier, D; Poole, S; Dantzer, R; Lestage, J

    2001-08-01

    Little is known on the forms of interleukin-1beta (IL-1beta) that are produced by microglial cells in the nervous system. Mixed glial cell cultures of rats produced IL-1beta in response to lipopolysaccharide (LPS). Using Western blot, pro-IL-1beta was found to be localized both intracellularly and in the supernatant, whereas mature IL-1beta was found only in the supernatant but in lower quantities than pro-IL-1beta. Immunocytochemistry confirmed that microglial cells are the exclusive source of IL-1beta. Blockade of the IL-1beta-converting enzyme (ICE) by Tyr-Val-Ala-Asp-aldehyde (YVAD-CHO) decreased the levels of mature IL-1beta but had no effect on pro-IL-1beta. Release of pro-IL-1beta was not associated with cell death nor with the extracellular release of ICE. Using gelatin zymography, glial cells were found to express constitutive matrix metalloproteinases (MMP) in the form of MMP-2. Exposure to LPS induced MMP-9 expression in a time-dependent manner similar to the pro-IL-1beta expression profile. MMP activation and inhibition experiments indicated a possible role of MMPs in the cleavage of pro-IL-1beta but not in the generation of mature IL-1beta. Microglial cells share with macrophages the ability to release large amounts of pro-IL-1beta of which the extracellular role remains to be determined. PMID:11556886

  7. Central injection of IL-10 antagonizes the behavioural effects of lipopolysaccharide in rats.

    Science.gov (United States)

    Bluthé, R M; Castanon, N; Pousset, F; Bristow, A; Ball, C; Lestage, J; Michaud, B; Kelley, K W; Dantzer, R

    1999-04-01

    Peripheral (i.p.) and central (i.c.v.) injections of lipopolysaccharide (LPS) have been shown to induce brain expression of proinflammatory cytokines and to depress social behaviour in rats, increase duration of immobility and induce body weight loss. To determine if the anti-inflammatory cytokine, interleukin-10 (IL-10) is able to modulate these effects, recombinant rat IL-10 was injected in the lateral ventricle of the brain (30, 100, 300 ng/rat) prior to i.p. or i.c.v. injection of LPS (250 micrograms/kg or 60 ng/rat, respectively). Social exploration was depressed for 6 h after i.p. LPS injection. This effect was attenuated by IL-10 (30 and 100 ng) 2 h after injection, whereas the highest dose of IL-10 blocked the depression of social interaction for 6 h after LPS injection. IL-10 produced the same effects on the increase of immobility although the results did not reach significance. Social exploration was depressed 3 h after i.c.v. LPS injection, and this was accompanied by increased immobility. These effects were totally blocked by i.c.v. IL-10 (300 ng/rat). Rats lost body weight after i.c.v. LPS, and this effect was attenuated by i.c.v. IL-10. These results indicate that IL-10 is able to modulate the production and/or action of central proinflammatory cytokines. PMID:10101735

  8. A novel mechanism for inhibition of lipopolysaccharide-induced proinflammatory cytokine production by valproic acid.

    Science.gov (United States)

    Jambalganiin, Ulziisaikhan; Tsolmongyn, Bilegtsaikhan; Koide, Naoki; Odkhuu, Erdenezaya; Naiki, Yoshikazu; Komatsu, Takayuki; Yoshida, Tomoaki; Yokochi, Takashi

    2014-05-01

    The inhibitory effect of valproic acid (VPA) on lipopolysaccharide (LPS)-induced inflammatory response was studied by using mouse RAW 264.7 macrophage-like cells. VPA pretreatment attenuated LPS-induced phosphorylation of phosphatidylinositol 3-kinase (PI3K) and Akt, but not nuclear factor (NF)-?B and mitogen-activated protein kinases. VPA reduced phosphorylation of MDM2, an ubiquitin ligase and then prevented LPS-induced p53 degradation, followed by enhanced p53 expression. Moreover, p53 small interfering RNA (siRNA) abolished the inhibitory action of VPA on LPS-induced NF-?B p65 transcriptional activation and further LPS-induced tumor necrosis factor (TNF)-? and interleukin (IL)-6 production. VPA prevented LPS-induced degradation of phosphatase and tensin homologue deleted on chromosome ten (PTEN) and up-regulated the PTEN expression. Taken together, VPA was suggested to down-regulate LPS-induced NF-?B-dependent transcriptional activity via impaired PI3K/Akt/MDM2 activation and enhanced p53 expression. A detailed mechanism for inhibition of LPS-induced inflammatory response by VPA is discussed. PMID:24631367

  9. N-Lipidated Peptide Dimers: Effective Antibacterial Agents against Gram-Negative Pathogens through Lipopolysaccharide Permeabilization.

    Science.gov (United States)

    Koh, Jun-Jie; Lin, Huifen; Caroline, Vonny; Chew, Yu Siang; Pang, Li Mei; Aung, Thet Tun; Li, Jianguo; Lakshminarayanan, Rajamani; Tan, Donald T H; Verma, Chandra; Tan, Ai Ling; Beuerman, Roger W; Liu, Shouping

    2015-08-27

    Treating infections caused by multidrug-resistant Gram-negative pathogens is challenging, and there is concern regarding the toxicity of the most effective antimicrobials for Gram-negative pathogens. We hypothesized that conjugating a fatty acid moiety onto a peptide dimer could maximize the interaction with lipopolysaccharide (LPS) and facilitate the permeabilization of the LPS barrier, thereby improving potency against Gram-negative pathogens. We systematically designed a series of N-lipidated peptide dimers that are active against Gram-negative bacteria, including carbapenem-resistant Enterobacteriaceae (CRE). The optimized lipid length was 6-10 carbons. At these lipid lengths, the N-lipidated peptide dimers exhibited strong LPS permeabilization. Compound 23 exhibited synergy with select antibiotics in most of the combinations tested. 23 and 32 also displayed rapid bactericidal activity. Importantly, 23 and 32 were nonhemolytic at 10 mg/mL, with no cellular or in vivo toxicity. These characteristics suggest that these compounds can overcome the limitations of current Gram-negative-targeted antimicrobials such as polymyxin B. PMID:26214729

  10. Immunotherapy using lipopolysaccharide-stimulated bone marrow-derived dendritic cells to treat experimental autoimmune encephalomyelitis.

    Science.gov (United States)

    Zhou, F; Ciric, B; Zhang, G-X; Rostami, A

    2014-12-01

    Lipopolysaccharide (LPS) produced by Gram-negative bacteria induces tolerance and suppresses inflammatory responses in vivo; however, the mechanisms are poorly understood. In this study we show that LPS induces apoptosis of bone marrow-derived dendritic cells (DCs) and modulates phenotypes of DCs. LPS treatment up-regulates expression of tolerance-associated molecules such as CD205 and galectin-1, but down-regulates expression of Gr-1 and B220 on CD11c(+) DCs. Moreover, LPS treatment regulates the numbers of CD11c(+) CD8(+) , CD11c(+) CD11b(low) and CD11c(+) CD11b(hi) DCs, which perform different immune functions in vivo. Our data also demonstrated that intravenous transfer of LPS-treated DCs blocks experimental autoimmune encephalomyelitis (EAE) development and down-regulates expression of retinoic acid-related orphan receptor gamma t (ROR-?t), interleukin (IL)-17A, IL-17F, IL-21, IL-22 and interferon (IFN)-? in myelin oligodendrocyte glycoprotein (MOG)-primed CD4(+) T cells in the peripheral environment. These results suggest that LPS-induced apoptotic DCs may lead to generation of tolerogenic DCs and suppress the activity of MOG-stimulated effector CD4(+) T cells, thus inhibiting the development of EAE in vivo. Our results imply a potential mechanism of LPS-induced tolerance mediated by DCs and the possible use of LPS-induced apoptotic DCs to treat autoimmune diseases such as multiple sclerosis. PMID:25138204

  11. Determining the neuroprotective effects of dextromethorphan in lipopolysaccharide?stimulated BV2 microglia.

    Science.gov (United States)

    Cheng, Wenjing; Li, Yunhong; Hou, Xiaolin; Bai, Bin; Li, Fan; Ding, Feijia; Ma, Jiao; Zhang, Nan; Shen, Ying; Wang, Yin

    2015-02-01

    Microglial activation has been recognized as being vital in the pathogenesis of several neurodegenerative disorders. Therefore, the identification of therapeutic drugs to prevent microglial activation and thus protect against inflammation?mediated neuronal injury, is required. In the present study, dextromethorphan (DM), a compound widely used in antitussive remedies that has been demonstrated to possess neuroprotective effects, was shown to reduce proinflammatory mediator production in lipopolysaccharide (LPS)?stimulated BV2 mouse microglial cells. Western blot analysis revealed that DM markedly suppressed the activation of nuclear factor??B (NF?B), caspase?3 signaling and the expression of another inflammation?inducing factor, heat shock protein 60 (HSP60) and heat shock factor?1, induced by LPS in BV2 cells. Results from ELISA assay demonstrated that DM reduced the release of HSP60, nitric oxide (NO), inducible NO synthase, tumor necrosis factor??, interleukin (IL)?1? and IL?6 induced by LPS in BV2 microglia. These results were confirmed by immunofluorescence, suggesting that DM may exert a neuroprotective and anti?inflammatory effect by inhibiting microglial activation through the HSP60?NF?B signaling pathway. Therefore, DM may offer substantial therapeutic benefits in the treatment of neurodegenerative diseases that are accompanied by microglial activation. PMID:25351178

  12. Lancemaside A inhibits lipopolysaccharide-induced inflammation by targeting LPS/TLR4 complex.

    Science.gov (United States)

    Joh, Eun-Ha; Kim, Dong-Hyun

    2010-11-01

    In our previous study, lancemaside A isolated from Codonopsis lanceolata (family Campanulaceae) ameliorated colitis in mice. In this study, the anti-inflammatory effects of lancemaside A was investigated in lipopolysaccharide (LPS)-stimulated mice and their peritoneal macrophage cells. Lancemaside A suppressed the production of pro-inflammatory cytokines, TNF-? and IL-1?, in vitro and in vivo. Lancemaside A also down-regulated inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), as well as the inflammatory mediators, nitric oxide (NO), and PGE(2). Lancemaside A also inhibited the expression of IL-1 receptor-associated kinase-4 (IRAK-4), the phosphorylation of IKK-? and I?B-?, the nuclear translocation of NF-?B and the activation of mitogen-activated protein kinases in LPS-stimulated peritoneal macrophages. Furthermore, lancemaisde A inhibited the interaction between LPS and TLR4, as well as IRAK-4 expression in peritoneal macrophages. Based on these findings, lancemaside A expressed anti-inflammatory effects by regulating both the binding of LPS to TLR4 on macrophages. PMID:20665542

  13. Effectiveness of Brucella abortus lipopolysaccharide as an adjuvant for tuberculin PPD.

    Science.gov (United States)

    Jamalan, Mostafa; Ardestani, Susan Kaboudanian; Zeinali, Majid; Mosaveri, Nader; Mohammad Taheri, Mohammad

    2011-01-01

    Bacterial lipopolysaccharide (LPS) has T-helper 1 (Th1) immunostimulatory activities but because of toxicity and pyrogenicity cannot be used as an adjuvant. Brucella abortus LPS has less toxicity and no pyrogenic properties in comparison to other bacterial LPS. In the current study, the immunostimulatory properties of B. abortus LPS were evaluated for its adjuvant activity. Tuberculin purified protein derivative (PPD) from Mycobacterium tuberculosis was extracted and after anion-exchange chromatography on Q-sepharose column, two fractions (17 and 23), which dominantly contained 30- and 70-kDa antigens, were collected for immunological studies. BALB/c mice were immunized with four different antigen preparations (BCG, PPD, 17th and 23rd PPD fractions) along with complete Freund's adjuvant or B. abortus LPS. The T-cell immune response of mice was assessed by measurement of Th1-type cytokine (IFN-?) and Th2-type cytokines (IL-5 and IL-10) levels. Also, the humoral immunity was evaluated by measuring the specific IgG levels. Our results showed that immunization of mice with 17th PPD fraction along with B. abortus LPS can induce a Th1-type cytokine response characterized with a high IFN-?/IL-5 ratio, while immunization with PPD or 23rd PPD fraction along with the same adjuvant resulted to a mixed Th1/Th2-type cytokine response. PMID:20965746

  14. Interactions of lipopolysaccharide with lipid membranes, raft models - a solid state NMR study.

    Science.gov (United States)

    Ciesielski, Filip; Griffin, David C; Rittig, Michael; Moriyón, Ignacio; Bonev, Boyan B

    2013-08-01

    Lipopolysaccharide (LPS) is a major component of the external leaflet of bacterial outer membranes, key pro-inflammatory factor and an important mediator of host-pathogen interactions. In host cells it activates the complement along with a pro-inflammatory response via a TLR4-mediated signalling cascade and shows preference for cholesterol-containing membranes. Here, we use solid state (13)C and (31)P MAS NMR to investigate the interactions of LPS from three bacterial species, Brucella melitensis, Klebsiella pneumoniae and Escherichia coli, with mixed lipid membranes, raft models. All endotoxin types are found to be pyrophosphorylated and Klebsiellar LPS is phosphonylated, as well. Carbon-13 MAS NMR indicates an increase in lipid order in the presence of LPS. Longitudinal (31)P relaxation, providing a direct probe of LPS molecular and segmental mobility, reveals a significant reduction in (31)P T1 times and lower molecular mobility in the presence of ternary lipid mixtures. Along with the ordering effect on membrane lipid, this suggests a preferential partitioning of LPS into ordered bilayer sphingomyelin/cholesterol-rich domains. We hypothesise that this is an important evolutionary drive for the selection of GPI-anchored raft-associated LPS-binding proteins as a first line of response to membrane-associated LPS. PMID:23567915

  15. 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

  16. 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.

  17. Tristetraprolin Mediates Anti-Inflammatory Effects of Carbon Monoxide on Lipopolysaccharide-Induced Acute Lung Injury.

    Science.gov (United States)

    Joe, Yeonsoo; Kim, Seul-Ki; Chen, Yingqing; Yang, Jung Wook; Lee, Jeong-Hee; Cho, Gyeong Jae; Park, Jeong Woo; Chung, Hun Taeg

    2015-11-01

    Low-dose inhaled carbon monoxide is reported to suppress inflammatory responses and exhibit a therapeutic effect in models of lipopolysaccharide (LPS)-induced acute lung injury (ALI). However, the precise mechanism by which carbon monoxide confers protection against ALI is not clear. Tristetraprolin (TTP; official name ZFP36) exerts anti-inflammatory effects by enhancing decay of proinflammatory cytokine mRNAs. With the use of TTP knockout mice, we demonstrate here that the protection by carbon monoxide against LPS-induced ALI is mediated by TTP. Inhalation of carbon monoxide substantially increased the pulmonary expression of TTP. carbon monoxide markedly enhanced the decay of mRNA-encoding inflammatory cytokines, blocked the expression of inflammatory cytokines, and decreased tissue damage in LPS-treated lung tissue. Moreover, knockout of TTP abrogated the anti-inflammatory and tissue-protective effects of carbon monoxide in LPS-induced ALI. These results suggest that carbon monoxide-induced TTP mediates the protective effect of carbon monoxide against LPS-induced ALI by enhancing the decay of mRNA encoding proinflammatory cytokines. PMID:26348577

  18. 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. 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

  19. Bacterial lipopolysaccharide induces osteoclast formation in RAW 264.7 macrophage cells

    International Nuclear Information System (INIS)

    Lipopolysaccharide (LPS) is a potent bone resorbing factor. The effect of LPS on osteoclast formation was examined by using murine RAW 264.7 macrophage cells. LPS-induced the formation of multinucleated giant cells (MGC) in RAW 264.7 cells 3 days after the exposure. MGCs were positive for tartrate-resistant acid phosphatase (TRAP) activity. Further, MGC formed resorption pits on calcium-phosphate thin film that is a substrate for osteoclasts. Therefore, LPS was suggested to induce osteoclast formation in RAW 264.7 cells. LPS-induced osteoclast formation was abolished by anti-tumor necrosis factor (TNF)-? antibody, but not antibodies to macrophage-colony stimulating factor (M-CSF) and receptor activator of nuclear factor (NF)-?B ligand (RANKL). TNF-? might play a critical role in LPS-induced osteoclast formation in RAW 264.7 cells. Inhibitors of NF-?B and stress activated protein kinase (SAPK/JNK) prevented the LPS-induced osteoclast formation. The detailed mechanism of LPS-induced osteoclast formation is discussed

  20. The lipopolysaccharide-activated innate immune response network of the horseshoe crab

    Directory of Open Access Journals (Sweden)

    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

  1. 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

  2. Effects of propofol on damage of rat intestinal epithelial cells induced by heat stress and lipopolysaccharides

    Directory of Open Access Journals (Sweden)

    J. Tang

    2013-06-01

    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 treated 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.

  3. Commensal enteric bacteria lipopolysaccharide impairs host defense against disseminated Candida albicans fungal infection.

    Science.gov (United States)

    Jiang, T T; Chaturvedi, V; Ertelt, J M; Xin, L; Clark, D R; Kinder, J M; Way, S S

    2015-07-01

    Commensal enteric bacteria maintain systemic immune responsiveness that protects against disseminated or localized infection in extra-intestinal tissues caused by pathogenic microbes. However, as shifts in infection susceptibility after commensal bacteria eradication have primarily been probed using viruses, the broader applicability to other pathogen types remains undefined. In sharp contrast to diminished antiviral immunity, we show commensal bacteria eradication bolsters protection against disseminated Candida albicans fungal infection. Enhanced antifungal immunity reflects more robust systemic expansion of Ly6G(hi)Ly6C(int) neutrophils, and their mobilization into infected tissues among antibiotic-treated compared with commensal bacteria-replete control mice. Reciprocally, depletion of neutrophils from expanded levels or intestinal lipopolysaccharide reconstitution overrides the antifungal protective benefits conferred by commensal bacteria eradication. This discordance in antifungal compared with antiviral immunity highlights intrinsic differences in how commensal bacteria control responsiveness for specific immune cell subsets, because pathogen-specific CD8(+) T cells that protect against viruses were suppressed similarly after C. albicans and influenza A virus infection. Thus, positive calibration of antiviral immunity by commensal bacteria is counterbalanced by restrained activation of other immune components that confer antifungal immunity. PMID:25492473

  4. 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

  5. Protective effect of forsythiaside A on lipopolysaccharide/d-galactosamine-induced liver injury.

    Science.gov (United States)

    Pan, Chen-Wei; Zhou, Guang-Yao; Chen, Wei-Lai; Zhuge, Lu; Jin, Ling-Xiang; Zheng, Yi; Lin, Wei; Pan, Zhen-Zhen

    2015-05-01

    Forsythiaside A, an active constituent isolated from air-dried fruits of Forsythia suspensa, has been reported to have multiple pharmacological activities including anti-inflammatory, anti-oxidant, and antioxidant activities. In the present study, the hepatoprotective effect of forsythiaside A was investigated in lipopolysaccharide (LPS)/d-galactosamine (GalN)-induced acute liver injury in mice. Mice acute liver injury model was induced by LPS (50?g/kg)/GalN (800mg/kg). Forsythiaside A was administrated 1h prior to LPS/GalN exposure. The results showed that forsythiaside A attenuated hepatic pathological damage, malondialdehyde (MDA) content, and serum ALT, and AST levels induced by LPS/GalN. Moreover, forsythiaside A inhibited NF-?B activation, serum TNF-? and hepatic TNF-? levels induced by LPS/GalN. Furthermore, we found that forsythiaside A up-regulated the expression of Nrf2 and heme oxygenase-1. Our results showed that forsythiaside A protected against LPS/GalN-induced liver injury through activation of Nrf2 and inhibition of NF-?B activation. PMID:25797347

  6. Interactions between chensinin-1, a natural antimicrobial peptide derived from Rana chensinensis, and lipopolysaccharide.

    Science.gov (United States)

    Dong, Weibing; Sun, Yue; Shang, Dejing

    2015-12-01

    Lipopolysaccharide (LPS) plays a critical role in the pathogenesis of sepsis caused by gram-negative bacterial infections. Therefore, LPS-neutralizing molecules would have important clinical applications. Chensinin-1, a novel antimicrobial peptide with atypical structural features, was found in the skin secretions of the Chinese brown frog Rana chensinensis. To understand the role of LPS in the bacterial susceptibility to chensinin-1 and to investigate its anti-endotoxin effects, the interactions of chensinin-1 with LPS were investigated in this study using circular dichroism, in situ IR, isothermal titration calorimetry, and zeta potential. This study is the first to use in situ IR spectroscopy to evaluate the secondary structural changes of this peptide. The capacity of chensinin-1 to block the LPS-dependent cytokine secretion of macrophages was also investigated. Our results show that chensinin-1 can form ?-helical structures in LPS suspensions. LPS can affect the antimicrobial activity of chensinin-1, and chensinin-1 was able to mitigate the effects of LPS. These data may facilitate the development of antimicrobial peptides with potent antimicrobial and anti-endotoxin activities. © 2015 Wiley Periodicals, Inc. Biopolymers 103: 719-726, 2015. PMID:26340228

  7. Cross-protective Immunity Against Leptospirosis Elicited by a Live, Attenuated Lipopolysaccharide Mutant

    Science.gov (United States)

    Srikram, Amporn; Zhang, Kunkun; Bartpho, Thanatchaporn; Lo, Miranda; Hoke, David E.; Sermswan, Rasana W.; Adler, Ben

    2011-01-01

    Background. Leptospira species cause leptospirosis, a zoonotic disease found worldwide. Current vaccines against leptospirosis provide protection only against closely related serovars. Methods. We evaluated an attenuated transposon mutant of Leptospira interrogans serovar Manilae (M1352, defective in lipopolysaccharide biosynthesis) as a live vaccine against leptospirosis. Hamsters received a single dose of vaccine and were challenged with the homologous serovar (Manilae) and a serologically unrelated heterologous serovar (Pomona). Comparisons were made with killed vaccines. Potential cross-protective antigens against leptospirosis were investigated. Results. Live M1352 vaccine induced superior protection in hamsters against homologous challenge. The live vaccine also stimulated cross-protection against heterologous challenge, with 100% survival (live M1352) versus 40% survival (killed vaccine). Hamsters receiving either vaccine responded to the dominant membrane proteins LipL32 and LipL41. Hamsters receiving the live vaccine additionally recognized LA3961/OmpL36 (unknown function), Loa22 (OmpA family protein, recognized virulence factor), LA2372 (general secretory protein G), and LA1939 (hypothetical protein). Manilae LigA was recognized by M1352 vaccinates, whereas LipL36 was detected in Pomona. Conclusion. This study demonstrated that a live, attenuated vaccine can stimulate cross-protective immunity to L. interrogans and has identified antigens that potentially confer cross-protection against leptospirosis. PMID:21220775

  8. 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

  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. 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; Yang, Woong Mo

    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

  11. 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.

  12. 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.

  13. 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.

  14. Rough-Form Lipopolysaccharide Increases Apoptosis in Human CD4? and CD8? T Lymphocytes.

    Science.gov (United States)

    Nielsen, J S; Larsson, A; Ledet, T; Turina, M; Tønnesen, E; Krog, J

    2012-02-01

    Immunosuppression induced by lymphocyte apoptosis is considered an important factor in the pathogenesis of sepsis and has been demonstrated in both animal models of lipopolysaccharide (LPS)-induced endotoxemia and septic patients. As rough-form LPS (R-LPS) has recently 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 multicolour 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 with both S-LPS-stimulated and unstimulated cells. 7-Amino-actinomycin D 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 on B or NK cells. Our results indicate that R-LPS induces 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. PMID:21854408

  15. Cerium oxide nanoparticles inhibit lipopolysaccharide induced MAP kinase/NF-kB mediated severe sepsis

    Directory of Open Access Journals (Sweden)

    Vellaisamy Selvaraj

    2015-09-01

    Full Text Available The life threatening disease of sepsis is associated with high mortality. Septic patient survivability with currently available treatments has failed to improve. The purpose of this study was to evaluate whether lipopolysaccharide (LPS induced sepsis mortality and associated hepatic dysfunction can be prevented by cerium oxide nanoparticles (CeO2NPs treatment in male Sprague Dawley rats. Here we provide the information about the methods processing of raw data related to our study published in Biomaterials (Selvaraj et al., Biomaterials, 2015, In press and Data in Brief (Selvaraj et al., Data in Brief, 2015, In Press. The data present here provides confirmation of cerium oxide nanoparticle treatments ability to prevent the LPS induced sepsis associated changes in physiological, blood cell count, inflammatory protein and growth factors in vivo. In vitro assays investigation the treated of macrophages cells with different concentrations of cerium oxide nanoparticle demonstrate that concentration of cerium oxide nanoparticles below 1 µg/ml did not significantly influence cell survival as determined by the MTT assay.

  16. CdTe quantum dots as a novel biosensor for Serratia marcescens and Lipopolysaccharide.

    Science.gov (United States)

    Ebrahim, Sh; Reda, M; Hussien, A; Zayed, D

    2015-11-01

    The main objective of this work is to synthesize CdTe quantum dots (QDs) conjugated with Concanavalin A (Con A) as a novel biosensor to be selective and specific for the detection of Lipopolysaccharide (LPS). In addition, the conjugated CdTe QDs-Con A was used as fluorescence labels to capture Serratia marcescens bacteria through the recognition between CdTe QDs-Con A and LPS of S. marcescens. The appearance of the lattice plans in the high resolution transmission electron photograph indicated a high crystalline with an average size of 4-5nm for the CdTe QDs. The results showed that the relative fluorescence intensity of CdTe QDs-Con A decreased linearly with LPS concentration in the range from 10 to 90fg/mL and with correlation coefficient (R(2)) equal to 0.9713. LPS surrounding the S. marcescens bacteria was bound to the CdTe QDs-Con A and leads to quenching of PL intensity. It was found that a good linear relationship between the relative PL intensity and the logarithmic of cell population of S. marcescens in range from 1×10 to 1×10(6)CFU/mL at pH 7 with R(2) of 0.952 was established. PMID:26051643

  17. Omega-3 Fatty Acid Intervention Suppresses Lipopolysaccharide-Induced Inflammation and Weight Loss in Mice

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    Ying-Hua Liu

    2015-02-01

    Full Text Available Bacterial endotoxin lipopolysaccharide (LPS-induced sepsis is a critical medical condition, characterized by a severe systemic inflammation and rapid loss of muscle mass. Preventive and therapeutic strategies for this complex disease are still lacking. Here, we evaluated the effect of omega-3 (n-3 polyunsaturated fatty acid (PUFA intervention on LPS-challenged mice with respect to inflammation, body weight and the expression of Toll-like receptor 4 (TLR4 pathway components. LPS administration induced a dramatic loss of body weight within two days. Treatment with n-3 PUFA not only stopped loss of body weight but also gradually reversed it back to baseline levels within one week. Accordingly, the animals treated with n-3 PUFA exhibited markedly lower levels of inflammatory cytokines or markers in plasma and tissues, as well as down-regulation of TLR4 pathway components compared to animals without n-3 PUFA treatment or those treated with omega-6 PUFA. Our data demonstrate that n-3 PUFA intervention can suppress LPS-induced inflammation and weight loss via, at least in part, down-regulation of pro-inflammatory targets of the TLR4 signaling pathway, and highlight the therapeutic potential of n-3 PUFA in the management of sepsis.

  18. Renal neutrophil gelatinase associated lipocalin expression in lipopolysaccharide-induced acute kidney injury in the rat

    Directory of Open Access Journals (Sweden)

    Han Mei

    2012-06-01

    Full Text Available Abstract Background Neutrophil gelatinase associated lipocalin (NGAL is a highly predictive biomarker of acute kidney injury. To understand the role of NGAL in renal injury during sepsis, we investigated the temporal changes and biological sources of NGAL in a rat model of acute kidney injury, and explored the relationship between renal inflammation, humoral NGAL and NGAL expression during endotoxemia. Methods To induce acute renal injury, rats were treated with lipopolysaccharide (LPS, 3.5 mg/kg, ip, and the location of NGAL mRNA was evaluated by in situ hybridization. Quantitative RT-PCR was also used to determine the dynamic changes in NGAL, tumor necrosis factor ? (TNF? and interleukin (IL-6 mRNA expression 1, 3, 6, 12, and 24 hours following LPS treatment. The correlation among NGAL, TNF? and IL-6 was analyzed. Urinary and plasma NGAL (u/pNGAL levels were measured, and the relationship between humoral NGAL and NGAL expression in the kidney was investigated. Results Renal function was affected 3–12 hours after LPS. NGAL mRNA was significantly upregulated in tubular epithelia at the same time (P P P P Conclusions NGAL upregulation is sensitive to LPS-induced renal TNF? increase and injury, which are observed in the tubular epithelia. Urinary NGAL levels accurately reflect changes in NGAL in the kidney.

  19. Chemical Profiles and Protective Effect of Hedyotis diffusa Willd in Lipopolysaccharide-Induced Renal Inflammation Mice

    Science.gov (United States)

    Ye, Jian-Hong; Liu, Meng-Hua; Zhang, Xu-Lin; He, Jing-Yu

    2015-01-01

    Protective effect of Hedyotis diffusa (H. diffusa) Willd against lipopolysaccharide (LPS)-induced renal inflammation was evaluated by the productions of cytokines and chemokine, and the bioactive constituents of H. diffusa were detected by the ultra-fast liquid chromatography -diode array detector-quadrupole-time of flight mass spectrometry (UFLC-DAD-Q-TOF-MS/MS) method. As the results showed, water extract of H. diffusa (equal to 5.0 g/kg body weight) obviously protected renal tissues, significantly suppressed the productions of tumor necrosis factor-? (TNF-?), interleukin (IL)-1?, IL-6, and monocyte chemoattractant protein (MCP)-1, as well as significantly promoted the production of IL-10 in serum and renal tissues. According the chemical profiles of H. diffusa, flavonoids, iridoid glycosides and anthraquinones were greatly detected in serum from H. diffusa extract treatment mice. Two main chemotypes, including eight flavonoids and four iridoid glycosides were found in renal tissues from H. diffusa extract treatment mice. The results demonstrated that water extract of H. diffusa had protective effect on renal inflammation, which possibly resulted from the bioactive constituents consisting of flavonoids, iridoids and anthraquinones. PMID:26580602

  20. 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.

  1. Upregulation of prolylcarboxypeptidase (PRCP in lipopolysaccharide (LPS treated endothelium promotes inflammation

    Directory of Open Access Journals (Sweden)

    Kolte Dhaval

    2009-01-01

    Full Text Available Abstract Background Prolylcarboxypeptidase (Prcp gene, along with altered PRCP and kallikrein levels, have been implicated in inflammation pathogenesis. PRCP regulates angiotensin 1–7 (Ang 1–7 – and bradykinin (BK – stimulated nitric oxide production in endothelial cells. The mechanism through which kallikrein expression is altered during infection is not fully understood. Investigations were performed to determine the association between PRCP and kallikrein levels as a function of the upregulation of PRCP expression and the link between PRCP and inflammation risk in lipopolysaccharide (LPS-induced endothelium activation. Methods The Prcp transcript expression in LPS-induced human umbilical vein endothelial cells (HUVEC activation was determined by RT-PCR for mRNA. PRCP-dependent kallikrein pathway was determined either by Enzyme Linked ImmunoSorbent Assay (ELISA or by biochemical assay. Results We report that PRCP is critical to the maintenance of the endothelial cells, and its upregulation contributes to the risk of developing inflammation. Significant elevation in kallikrein was seen on LPS-treated HUVECs. The conversion of PK to kallikrein was blocked by the inhibitor of PRCP, suggesting that PRCP might be a risk factor for inflammation. Conclusion The increased PRCP lead to a sustained production of bradykinin in endothelium following LPS treatment. This amplification may be an additional mechanism whereby PRCP promotes a sustained inflammatory response. A better appreciation of the role of PRCP in endothelium may contribute to a better understanding of inflammatory vascular disorders and to the development of a novel treatment.

  2. pVEGF-loaded lipopolysaccharide-amine nanopolymersomes for therapeutic angiogenesis

    Science.gov (United States)

    Teng, Wei; Huang, Zhonghui; Chen, Ying; Wang, Lichun; Wang, Qinmei; Huang, Hongzhang

    2014-02-01

    Therapeutic angiogenesis via gene delivery is promising for tissue survival and regeneration after injury or ischemia. A stable, safe and efficient gene vector is essential for successful angiogenesis. We have demonstrated that our newly developed lipopolysaccharide-amine nanopolymersomes (LNPs) have higher than 95% transfection efficiency when delivering pEGFP into mesenchymal stem cells (MSCs). To explore their clinical potential in therapeutic angiogenesis, in this study, we studied their toxicity, storage stability, protection ability to genes and efficacy to deliver therapeutic genes of pVEGF in MSCs and zebrafish. The results show that LNPs can condense pVEGF to form pVEGF-loaded nanopolymersomes (VNPs), and protect pVEGF against DNase digestion in 6 h. Both LNPs and VNPs have low toxicity to MSCs, erythrocytes and zebrafish embryos. LNPs are stable at 4?°C for at least two years with unchanged size and transfection efficiency. MSCs transfected by VNPs continuously synthesize VEGF for at least four days under control, with a peak (21.25 ng ml-1) ˜35-fold greater than that for the untreated group. VNPs induce significant and dose-dependent angiogenesis in zebrafish without causing death, deformity or delay in growth and development, and the induced maximal vessel area of subintestinal vessel plexus is 2.5-fold higher than that for the untreated group. Our study suggests that VNP has high potential in therapeutic angiogenesis.

  3. Withaferin A attenuates lipopolysaccharide-induced acute lung injury in neonatal rats.

    Science.gov (United States)

    Gao, S; Li, H; Zhou, X-Q; You, J-B; Tu, D-N; Xia, G; Jiang, J-X; Xin, C

    2015-01-01

    Withaferin A (WFA) is an active compound from Withania somnifera and has been reported to exhibit a variety of pharmacological activities such as anti—inflammatory, immunomodulatory and anti—tumor properties. In the present study, we investigated the potential protective role of WFA on acute lung injury in neonatal rats induced by lipopolysaccharide (LPS). We found that WFA significantly attenuated the pathological changes of lungs induced by LPS injection. Administration with WFA obviously decreased pulmonary neutrophil infiltration accompanied with decreased MPO concentrations. WFA also reduced the expression of pro—inflammatory cytokines including MIP—2, TNF—?, IL—1? and IL—6. Meanwhile, the expression levels of anti—inflammatory mediators such as TGF—?1 and IL—10 were significantly increased following WFA administration. Moreover, WFA protected LPS—treated rats from oxidative damage via up—regulation of TBARS and H2O2 concentrations and down—regulation of ROS contents. Taken together, the present study demonstrated that WFA administration attenuated LPS—induced lung injury through inhibition of inflammatory responses and oxidative stress. PMID:26255139

  4. 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

  5. Role of Cell Surface Lipopolysaccharides in Escherichia coli K12 adhesion and transport.

    Science.gov (United States)

    Walker, Sharon L; Redman, Jeremy A; Elimelech, Menachem

    2004-08-31

    The influence of bacterial surface lipopolysaccharides (LPS) on cell transport and adhesion has been examined by use of three mutants of Escherichia coli K12 with well-characterized LPS of different lengths and molecular composition. Two experimental techniques, a packed-bed column and a radial stagnation point flow system, were employed to investigate bacterial adhesion kinetics onto quartz surfaces over a wide range of solution ionic strengths. Although the two systems capture distinct deposition (adhesion) mechanisms because of their different hydrodynamics, similar deposition kinetics trends were observed for each bacterial strain. Bacterial deposition rates were directly related to the electrostatic double layer interaction between the bacteria and quartz surfaces, in qualitative agreement with classic Derjaguin-Landau-Verwey-Overbeek (DLVO) theory. However, DLVO theory does not fully explain the deposition behavior for the bacterial strain with the lengthy, uncharged O-antigen portion of the LPS. Neither the length nor the charge characteristics of the LPS molecule directly correlated to deposition kinetics, suggesting a complex combination of cell surface charge heterogeneity and LPS composition controls the bacterial adhesive characteristics. It is further suggested that bacterial deposition behavior is determined by the combined influence of DLVO interactions, LPS-associated chemical interactions, and the hydrodynamics of the deposition system. PMID:15323526

  6. 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

  7. 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.

  8. 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.

  9. Imaging robust microglial activation after lipopolysaccharide administration in humans with PET.

    Science.gov (United States)

    Sandiego, Christine M; Gallezot, Jean-Dominique; Pittman, Brian; Nabulsi, Nabeel; Lim, Keunpoong; Lin, Shu-Fei; Matuskey, David; Lee, Jae-Yun; O'Connor, Kevin C; Huang, Yiyun; Carson, Richard E; Hannestad, Jonas; Cosgrove, Kelly P

    2015-10-01

    Neuroinflammation is associated with a broad spectrum of neurodegenerative and psychiatric diseases. The core process in neuroinflammation is activation of microglia, the innate immune cells of the brain. We measured the neuroinflammatory response produced by a systemic administration of the Escherichia coli lipopolysaccharide (LPS; also called endotoxin) in humans with the positron emission tomography (PET) radiotracer [(11)C]PBR28, which binds to translocator protein, a molecular marker that is up-regulated by microglial activation. In addition, inflammatory cytokines in serum and sickness behavior profiles were measured before and after LPS administration to relate brain microglial activation with systemic inflammation and behavior. Eight healthy male subjects each had two 120-min [(11)C]PBR28 PET scans in 1 d, before and after an LPS challenge. LPS (1.0 ng/kg, i.v.) was administered 180 min before the second [(11)C]PBR28 scan. LPS administration significantly increased [(11)C]PBR28 binding 30-60%, demonstrating microglial activation throughout the brain. This increase was accompanied by an increase in blood levels of inflammatory cytokines, vital sign changes, and sickness symptoms, well-established consequences of LPS administration. To our knowledge, this is the first demonstration in humans that a systemic LPS challenge induces robust increases in microglial activation in the brain. This imaging paradigm to measure brain microglial activation with [(11)C]PBR28 PET provides an approach to test new medications in humans for their putative antiinflammatory effects. PMID:26385967

  10. 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.

  11. Lipopolysaccharide-induced multinuclear cells: Increased internalization of polystyrene beads and possible signals for cell fusion

    International Nuclear Information System (INIS)

    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. Subclinical decelerations during developing hypotension in preterm fetal sheep after acute on chronic lipopolysaccharide exposure.

    Science.gov (United States)

    Lear, Christopher A; Davidson, Joanne O; Galinsky, Robert; Yuill, Caroline A; Wassink, Guido; Booth, Lindsea C; Drury, Paul P; Bennet, Laura; Gunn, Alistair J

    2015-01-01

    Subclinical (shallow) heart rate decelerations occur during neonatal sepsis, but there is limited information on their relationship with hypotension or whether they occur before birth. We examined whether subclinical decelerations, a fall in fetal heart rate (FHR) that remained above 100?bpm, were associated with hypotension in preterm fetal sheep exposed to lipopolysaccharide (LPS). Chronically-instrumented fetal sheep at 0.7 gestation received continuous low-dose LPS infusions (n?=?15, 100?ng/kg over 24?h, followed by 250?ng/kg/24?h for 96?h) or saline (n?=?8). Boluses of 1??g LPS or saline were given at 48 and 72?h. FHR variability (FHRV) was calculated, and sample asymmetry was used to assess the severity and frequency of decelerations. Low-dose LPS infusion did not affect FHR. After the first LPS bolus, 7 fetuses remained normotensive, while 8 developed hypotension (a fall in mean arterial blood pressure of ?5?mmHg). Developing hypotension was associated with subclinical decelerations, with a corresponding increase in sample asymmetry and FHRV (p?

  13. 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.

  14. Expression of the lipopolysaccharide biosynthesis gene lpxD affects biofilm formation of Pseudomonas aeruginosa.

    Science.gov (United States)

    Alshalchi, Sahar A; Anderson, Gregory G

    2015-03-01

    Bacterial biofilms are an important cause of nosocomial infections. Microorganisms such as Pseudomonas aeruginosa colonize biotic and abiotic surfaces leading to chronic infections that are difficult to eradicate. To characterize novel genes involved in biofilm formation, we identified the lpxD gene from a transposon-mutant library of P. aeruginosa. This gene encodes a glucosamine-N acyltransferase, which is important for lipopolysaccharide biosynthesis. Our results showed that a loss-of-expression mutant of lpxD was defective for biofilm formation on biotic and abiotic surfaces. Additionally, this mutant strain exhibited significantly decreased bacterial attachment to cultured airway epithelial cells, as well as increased bacterial cytotoxicity toward airway cells. However, consistent with a defect in lipid A structure, airway cells incubated with the lpxD mutant or with mutant lipid A extracts exhibited decreased IL-8 production and necrosis, respectively. Overall, our data indicate that manipulating lpxD expression may influence P. aeruginosa's ability to establish biofilm infections. PMID:25173672

  15. Effect of arazyme on the lipopolysaccharide?induced inflammatory response in human endothelial cells.

    Science.gov (United States)

    Kim, In Sik; Yang, Eun Ju; Shin, Dong-Ha; Son, Kwang-Hee; Park, Ho-Yong; Lee, Ji-Sook

    2014-08-01

    Arazyme is a novel extracellular metalloprotease secreted by Aranicola proteolyticus. Endothelial cells are involved in the pathogenesis of a number of inflammatory diseases, induce uncontrolled cell viability and express various inflammatory mediators, including cytokines, chemokines, adhesion molecules and reactive oxygen species (ROS). In the current study, human umbilical vein endothelilal cells (HUVECs) were used to investigate the anti?inflammatory effects of arazyme following lipopolysaccharide (LPS) stimulation. Apoptosis of HUVECs due to LPS was inhibited by arazyme. In various inflammatory responses induced by LPS, arazyme inhibited the secretion of the monocyte chemoattractant protein?1 and interleukin?6, and the expression of vascular cell adhesion molecule?1 and intercellular adhesion molecule?1. Arazyme also suppressed ROS production in HUVECs. The action of arazyme was not associated with NF??B activity in HUVECs. These results indicate that arazyme has anti?inflammatory properties in inflamed endothelial cells and may be useful as a therapeutic agent for inflammatory diseases associated with endothelial cells. PMID:24842222

  16. Metformin suppresses intrahepatic coagulation activation in mice with lipopolysaccharide/D?galactosamine?induced fulminant hepatitis.

    Science.gov (United States)

    Gong, Xianqiong; Duan, Rui; Ao, Jin-E; Ai, Qing; Ge, Pu; Lin, Ling; Zhang, Li

    2015-10-01

    Metformin is a widely?used antidiabetic drug with hypoglycemic activity and previously described anti?inflammatory properties. Previous studies have demonstrated that metformin attenuates endotoxic hepatitis, however the mechanisms remain unclear. Inflammation and coagulation are closely associated pathological processes, therefore the potential effects of metformin on key steps in activation of the coagulation system were further investigated in endotoxic hepatitis induced by lipopolysaccharide/D?galactosamine (LPS/D?Gal). The current study demonstrated that treatment with metformin significantly suppressed the upregulation of tissue factor and plasminogen activator inhibitor?1 in LPS/D?Gal?exposed mice. In addition, a reduction in the expression of interleukin 6 and inhibition of nuclear translocation of nuclear factor??B were observed. These data indicate that the LPS/D?Gal?induced elevation of the stable protein level of hypoxia inducible factor 1?, the mRNA level of erythropoietin, vascular endothelial growth factor and matrix metalloproteinase?3, and the hepatic level of lactic acid were also suppressed by metformin. The current study indicates that the suppressive effects of metformin on inflammation?induced coagulation may be an additional mechanism underlying the hepatoprotective effects of metformin in mice with LPS/D?Gal?induced fulminant hepatitis. PMID:26260849

  17. Extraction and separation of lipopolysaccharide from outer membrane of Helicobacter pylori

    Directory of Open Access Journals (Sweden)

    Zavarreza J

    2007-06-01

    Full Text Available Background: Helicobacter pylori (H. pylori is one of the major causes of peptic ulcer, gastritis and gastric cancer. This bacterium has a special lipopolysaccharide (LPS, which is responsible for its pathogenesis and its high resistance against gastric acid and escape from the human immune system. This property makes it a target for further research and diagnostic goals. In this study, the extraction of the LPS and separation from the outer membrane is required. Methods: The LPS was extracted from the outer membrane, or envelope, of H. pylori obtained from patients suffering from gastritis, gastric ulcer and gastric cancer. LPS extraction was performed using the proteinase K method. SDS-PAGE and silver staining were applied to investigate the electrophoretic pattern of the LPS. This pattern was compared with that of E. coli serotype O111:B4 and Salmonella serotype ATCC 14028. Results: The extracted LPS has a ladder-shaped electrophoretic pattern and the bands are located in three groups: high, medium and low molecular weights. Conclusion: The distribution of the bands of the ladder-shaped electrophoretic pattern is caused by the different number of oligosaccharide chains associated with the LPS. The high molecular weight bands represent S-LPS and the low molecular weight bands represent the R-LPS, which lacks the O-chain.

  18. Maternal molecular hydrogen administration on lipopolysaccharide-induced mouse fetal brain injury

    Science.gov (United States)

    Nakano, Tomoko; Kotani, Tomomi; Mano, Yukio; Tsuda, Hiroyuki; Imai, Kenji; Ushida, Takafumi; Li, Hua; Miki, Rika; Sumigama, Seiji; Sato, Yoshiaki; Iwase, Akira; Hirakawa, Akihiro; Asai, Masato; Toyokuni, Shinya; Kikkawa, Fumitaka

    2015-01-01

    Fetal brain injury is often related to prenatal inflammation; however, there is a lack of effective therapy. Recently, molecular hydrogen (H2), a specific antioxidant to hydroxyl radical and peroxynitrite, has been reported to have anti-inflammatory properties. The aim of this study was to investigate whether maternal H2 administration could protect the fetal brain against inflammation. Pregnant C3H/HeN mice received an intraperitoneal injection of lipopolysaccharide (LPS) on gestational day 15.5 and were provided with H2 water for 24 h prior to LPS injection. Pup brain samples were collected on gestational day 16.5, and the levels of apoptosis and oxidative damage were evaluated using immunohistochemistry. Interleukin-6 (IL-6) levels were examined using real-time PCR. The levels of apoptosis and oxidative damage, as well as the levels of IL-6 mRNA, increased significantly when the mother was injected with LPS than that in the control group. However, these levels were significantly reduced when H2 was administered prior to the LPS-injection. Our results suggest that LPS-induced apoptosis, oxidative damage and inflammation in the fetal brain were ameliorated by maternal H2 administration. Antenatal H2 administration might protect the premature brain against maternal inflammation. PMID:26566302

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

    Directory of Open Access Journals (Sweden)

    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.

  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. Enzyme-linked immunosorbent assay for detection of Salmonella lipopolysaccharide in poultry specimens.

    Science.gov (United States)

    Rigby, C E

    1984-01-01

    An enzyme-linked immunosorbent assay (ELISA) for detection of salmonellae was developed and evaluated by using artificially contaminated specimens of poultry feed, feces, litter, or carcass rinsings, and naturally contaminated water samples. Specimens containing salmonellae of serogroups B or C2 inhibited the binding of polyvalent anti-O serum to microtiter plate wells coated with lipopolysaccharide of Salmonella typhimurium (serogroup B) or Salmonella albany (serogroup C2), respectively. Treatment of specimens with Rhozyme 41 (a protease) inhibited nonspecific reactions. The ELISA detected 106 of 111 culture-positive specimens contaminated with salmonellae of serogroups B or C2. Nineteen of 20 specimens containing salmonellae of serogroup C1 and all of 36 culture-negative specimens were ELISA negative. All seven water samples that contained salmonellae of serogroups B or C2, including three that were culture positive only after delayed secondary enrichment, were ELISA positive. Seven of the nine water samples that contained salmonellae of other serogroups, and all 38 culture-negative samples, were ELISA negative. The ELISA was simple to perform, produced results in 48 h, and was more economical than culture methods. PMID:6378094

  2. Neuroprotective effects of Lycium barbarum polysaccharides in lipopolysaccharide-induced BV2 microglial cells.

    Science.gov (United States)

    Teng, Peng; Li, Yunhong; Cheng, Wenjing; Zhou, Liang; Shen, Ying; Wang, Yin

    2013-06-01

    Polysaccharides extracted from Lycium barbarum (LBPs) possess a wide variety of biological activities. However, their neuroprotective effects have not yet been fully elucidated. The aim of the present study was to investigate the inhibitory effects of LBPs on the production of lipopolysaccharide (LPS)?induced proinflammatory mediators in BV2 microglia. BV2 mouse microglial cells were cultured and an MTT assay was performed to determine whether LBPs had an effect on the apoptosis of LPS-stimulated BV2 cells. Our data showed that LPS induced the activation of nuclear factor??B (NF??B) and its upstream protein caspase 3, upregulated the expression of an additional apoptosis?inducing factor, heat shock protein 60 (HSP60), in BV2 microglial cells and increased the release of TNF-? and HSP60 in the culture media. Following treatment with LBPs, the activated NF??B and caspase 3 were significantly suppressed. Furthermore, the enhanced expression of HSP60 was reduced and the LPS-induced release of TNF-? and HSP60 were inhibited. These results suggest that LBPs may have therapeutic potential for the treatment of neurodegenerative diseases that are accompanied by microglial activation. PMID:23620217

  3. Lipopolysaccharide aggregates in native agarose gels detected by reversible negative staining with imidazole and zinc salts.

    Science.gov (United States)

    Rodríguez, Caridad; Hardy, Eugenio

    2015-09-15

    We investigated the use of imidazole and zinc salts for the detection of lipopolysaccharide (LPS) aggregates separated by native agarose gel electrophoresis (NAGE). As a result, a new staining procedure was established by which as little as 1.5 ?g of Escherichia coli O55:B5 LPS aggregates was detected by means of inducing a clear, transparent pattern, contrasted against an opaque background. E. coli O55:B5 LPS preparations treated with nucleases and proteinase K proved that the reverse-stained LPS pattern is not related to any potential artifacts caused by unrelated biomolecules (e.g., nucleic acids, proteins). After this, we showed that the procedure is applicable to two-dimensional LPS separation using NAGE/SDS-PAGE, while at the same time confirming that real polydisperse LPS aggregates are represented by the stained profile. Also, we demonstrated the general applicability of this stain to the detection of different NAGE-separated LPS aggregates (e.g., from E. coli 026:B6, E. coli 0111:B4, Salmonella minnesota Re595). Finally, using lysozyme as a model protein, we found that imidazole-zinc may be combined with Coomassie brilliant blue R-250 into a double-staining process to enable the use of NAGE for investigating the interaction of cationic proteins and LPS aggregates and protein or LPS concentration effects on protein-LPS binding. PMID:26095395

  4. 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.

  5. 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

  6. 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

  7. 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.

  8. Inhibitory effects of French pine bark extract, Pycnogenol®, on alveolar bone resorption and on the osteoclast differentiation.

    Science.gov (United States)

    Sugimoto, Hideki; Watanabe, Kiyoko; Toyama, Toshizo; Takahashi, Shun-suke; Sugiyama, Shuta; Lee, Masaichi-Chang-il; Hamada, Nobushiro

    2015-02-01

    Pycnogenol(®) (PYC) is a standardized bark extract from French maritime pine (Pinus pinaster Aiton). We examined the inhibitory effects of PYC on alveolar bone resorption, which is a characteristic feature of periodontitis, induced by Porphyromonas gingivalis (P.?gingivalis) and osteoclast differentiation. In rat periodontitis model, rats were divided into four groups: group A served as the non-infected control, group B was infected orally with P.?gingivalis ATCC 33277, group C was administered PYC in the diet (0.025%: w/w), and group D was infected with P.?gingivalis and administered PYC. Administration of PYC along with P.?gingivalis infection significantly reduced alveolar bone resorption. Treatment of P.?gingivalis with 1?µg/ml PYC reduced the number of viable bacterial cells. Addition of PYC to epithelial cells inhibited adhesion and invasion by P.?gingivalis. The effect of PYC on osteoclast formation was confirmed by tartrate-resistant acid phosphatase staining. PYC treatment significantly inhibited osteoclast formation. Addition of PYC (1-100?µg/ml) to purified osteoclasts culture induced cell apoptosis. These results suggest that PYC may prevent alveolar bone resorption through its antibacterial activity against P.?gingivalis and by suppressing osteoclastogenesis. Therefore, PYC may be useful as a therapeutic and preventative agent for bone diseases such as periodontitis. PMID:25336411

  9. 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 mellem observatørerne 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 m

  10. Exogenous normal lymph reduces liver injury induced by lipopolysaccharides in rats

    Scientific Electronic Library Online (English)

    Z.G., Zhao; L.L., Zhang; C.Y., Niu; J., Zhang.

    2014-02-01

    Full Text Available The liver is one of the target organs damaged by septic shock, wherein the spread of endotoxins begins. This study aimed to investigate the effects of exogenous normal lymph (ENL) on lipopolysaccharide (LPS)-induced liver injury in rats. Male Wistar rats were randomly divided into sham, LPS, and LPS [...] +ENL groups. LPS (15 mg/kg) was administered intravenously via the left jugular vein to the LPS and LPS+ENL groups. At 15 min after the LPS injection, saline or ENL without cell components (5 mL/kg) was administered to the LPS and LPS+ENL groups, respectively, at a rate of 0.5 mL/min. Hepatocellular injury indices and hepatic histomorphology, as well as levels of P-selectin, intercellular adhesion molecule 1 (ICAM-1), myeloperoxidase (MPO), and Na+-K+-ATPase, were assessed in hepatic tissues. Liver tissue damage occurred after LPS injection. All levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in plasma as well as the wet/dry weight ratio of hepatic tissue in plasma increased. Similarly, P-selectin, ICAM-1, and MPO levels in hepatic tissues were elevated, whereas Na+-K+-ATPase activity in hepatocytes decreased. ENL treatment lessened hepatic tissue damage and decreased levels of AST, ALT, ICAM-1, and MPO. Meanwhile, the treatment increased the activity of Na+-K+-ATPase. These results indicated that ENL could alleviate LPS-induced liver injury, thereby suggesting an alternative therapeutic strategy for the treatment of liver injury accompanied by severe infection or sepsis.

  11. 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.

  12. Valproic acid potentiates curcumin-mediated neuroprotection in Lipopolysaccharide induced rats

    Directory of Open Access Journals (Sweden)

    Amira Zaky

    2014-10-01

    Full Text Available The etiology of neuroinflammation is complex and comprises multifactorial, involving both genetic and environmental factors during which diverse genetic and epigenetic modulations are implicated. Curcumin (Cur, and valproic acid (VPA, histone deacetylase 1 inhibitor, have neuroprotective effects. The present study was designed with an aim to investigate the ability of co-treatment of both compounds (Cur or VPA (200mg/kg for four weeks to augment neuroprotection and enhance brain recovery from intra-peritoneal (IP injection of (250 µg/kg lipopolysaccharide (LPS-stimulated neuroinflammatory condition on rat brain cortex. Cortex activation and the effects of combined treatment and production of proinflammatory mediators, COX-2, APE1 and nitric oxide/iNOS were investigated. Neuroinflammation development was assessed by histological analyses and by investigating associated indices (BACE1, APP, PSEN-1 and PSEN-2. Furthermore we measured the expression profile of let-7 miRNAs members a, b, c, e and f in all groups, a highly abundant regulator of gene expression in the CNS. Protein and mRNA levels of neuroinflammation markers COX-2, BACE1, APP and iNOS were also attenuated by combined therapy. On the other hand, assessment of the indicated five let-7 members, showed distinct expression profile pattern in the different groups. Let-7 a, b and c disappeared in the induced group, an effect that was partially suppressed by co-addition of either Cur or VPA. These data suggest that the combined treatment induced significantly the expression of the five members when compared to rats treated with Cur or VPA only as well as to self-recovery group, which indicates a possible benefit from the synergistic effect of Cur-VPA combination as therapeutic agents for neuroinflammation and its associated disorders. The mechanism elucidated here highlights the particular drug-induced expression profile of let-7 family as new targets for future pharmacological development.

  13. OPTICAL IMAGING OF LIPOPOLYSACCHARIDE-INDUCED OXIDATIVE STRESS IN ACUTE LUNG INJURY FROM HYPEROXIA AND SEPSIS.

    Science.gov (United States)

    Sepehr, Reyhaneh; Audi, Said H; Maleki, Sepideh; Staniszewski, Kevin; Eis, Annie L; Konduri, Girija G; Ranji, Mahsa

    2013-07-01

    Reactive oxygen species (ROS) have been implicated in the pathogenesis of many acute and chronic pulmonary disorders such as acute lung injury (ALI) in adults and bronchopulmonary dysplasia (BPD) in premature infants. Bacterial infection and oxygen toxicity, which result in pulmonary vascular endothelial injury, contribute to impaired vascular growth and alveolar simplification seen in the lungs of premature infants with BPD. Hyperoxia induces ALI, reduces cell proliferation, causes DNA damage and promotes cell death by causing mitochondrial dysfunction. The objective of this study was to use an optical imaging technique to evaluate the variations in fluorescence intensities of the auto-fluorescent mitochondrial metabolic coenzymes, NADH and FAD in four different groups of rats. The ratio of these fluorescence signals (NADH/FAD), referred to as NADH redox ratio (NADH RR) has been used as an indicator of tissue metabolism in injuries. Here, we investigated whether the changes in metabolic state can be used as a marker of oxidative stress caused by hyperoxia and bacterial lipopolysaccharide (LPS) exposure in neonatal rat lungs. We examined the tissue redox states of lungs from four groups of rat pups: normoxic (21% O2) pups, hyperoxic (90% O2) pups, pups treated with LPS (normoxic + LPS), and pups treated with LPS and hyperoxia (hyperoxic + LPS). Our results show that hyperoxia oxidized the respiratory chain as reflected by a ~31% decrease in lung tissue NADH RR as compared to that for normoxic lungs. LPS treatment alone or with hyperoxia had no significant effect on lung tissue NADH RR as compared to that for normoxic or hyperoxic lungs, respectively. Thus, NADH RR serves as a quantitative marker of oxidative stress level in lung injury caused by two clinically important conditions: hyperoxia and LPS exposure. PMID:24672581

  14. Extrusion of mitochondrial contents from lipopolysaccharide-stimulated cells: Involvement of autophagy.

    Science.gov (United States)

    Unuma, Kana; Aki, Toshihiko; Funakoshi, Takeshi; Hashimoto, Kyoko; Uemura, Koichi

    2015-09-01

    Sepsis/endotoxemia is elicited by the circulatory distribution of pathogens/endotoxins into whole bodies, and causes profound effects on human health by causing inflammation in multiple organs. Mitochondrial damage is one of the characteristics of the cellular degeneration observed during sepsis/endotoxemia. Elimination of damaged mitochondria through the autophagy-lysosome system has been reported in the liver, indicating that autophagy should play an important role in liver homeostasis during sepsis/endotoxemia. An increased appearance of mitochondrial DNA and proteins in the plasma is another feature of sepsis/endotoxemia, suggesting that damaged mitochondria are not only eliminated within the cells, but also extruded through currently unknown mechanisms. Here we provide evidence for the secretion of mitochondrial proteins and DNA from lipopolysaccharide (LPS)-stimulated rat hepatocytes as well as mouse embryonic fibroblasts (MEFs). The secretion of mitochondrial contents is accompanied by the secretion of proteins that reside in the lumenal space of autolysosomes (LC3-II and CTSD/cathepsin D), but not by a lysosomal membrane protein (LAMP1). The pharmacological inhibition of autophagy by 3MA blocks the secretion of mitochondrial constituents from LPS-stimulated hepatocytes. LPS also stimulates the secretion of mitochondrial as well as autolysosomal lumenal proteins from wild-type (Atg5(+/+)) MEFs, but not from atg5(-/-) MEFs. Furthermore, we show that direct exposure of purified mitochondria activates polymorphonuclear leukocytes (PMNs), as evident by the induction of IL1B/interlekin-1?, a pro-inflammatory cytokine. Taken together, the data suggest the active extrusion of mitochondrial contents, which provoke an inflammatory response of immune cells, through the exocytosis of autolysosomes by cells stimulated with LPS. PMID:26102061

  15. Metabolic and Hematological Consequences of Dietary Deoxynivalenol Interacting with Systemic Escherichia coli Lipopolysaccharide

    Directory of Open Access Journals (Sweden)

    Erik Bannert

    2015-11-01

    Full Text Available Previous studies have shown that chronic oral deoxynivalenol (DON exposure modulated Escherichia coli lipopolysaccharide (LPS-induced systemic inflammation, whereby the liver was suspected to play an important role. Thus, a total of 41 barrows was fed one of two maize-based diets, either a DON-diet (4.59 mg DON/kg feed, n = 19 or a control diet (CON, n = 22. Pigs were equipped with indwelling catheters for pre- or post-hepatic (portal vs. jugular catheter infusion of either control (0.9% NaCl or LPS (7.5 µg/kg BW for 1h and frequent blood sampling. This design yielded six groups: CON_CONjugular?CONportal, CON_CONjugular?LPSportal, CON_LPSjugular?CONportal, DON_CONjugular?CONportal, DON_CONjugular?LPSportal and DON_LPSjugular?CONportal. Blood samples were analyzed for blood gases, electrolytes, glucose, pH, lactate and red hemogram. The red hemogram and electrolytes were not affected by DON and LPS. DON-feeding solely decreased portal glucose uptake (p < 0.05. LPS-decreased partial oxygen pressure (pO2 overall (p < 0.05, but reduced pCO2 only in arterial blood, and DON had no effect on either. Irrespective of catheter localization, LPS decreased pH and base-excess (p < 0.01, but increased lactate and anion-gap (p < 0.01, indicating an emerging lactic acidosis. Lactic acidosis was more pronounced in the group DON_LPSjugular-CONportal than in CON-fed counterparts (p < 0.05. DON-feeding aggravated the porcine acid-base balance in response to a subsequent immunostimulus dependent on its exposure site (pre- or post-hepatic.

  16. Stem cell intervention ameliorates epigallocatechin-3-gallate/lipopolysaccharide-induced hepatotoxicity in mice.

    Science.gov (United States)

    Saleh, I G; Ali, Z; Hammad, M A; Wilson, F D; Hamada, F M; Abd-Ellah, M F; Walker, L A; Khan, I A; Ashfaq, M K

    2015-11-01

    Stem cells are identified as a novel cell therapy for regenerative medicine because of their ability to differentiate into many functional cell types. We have shown earlier a new model of hepatotoxicity in mice by administering (1500 mg/kg) epigallocatechin-3-gallate (EGCG) intragastric (IG) for 5 days after a single intraperitoneal dose (6 mg/kg) of lipopolysaccharide (LPS). In this study, we aimed to study the effect of intrahepatic (IH) injection of mouse embryonic stem cells (MESCs) on the hepatotoxicity induced by EGCG/LPS in mice. Mice were administered EGCG/LPS and rested for 3 days. MESCs were obtained from American Type Culture Collection and cultured in vitro for 4 days. Stem cells were injected IH. Seven days later, a single dose of LPS (6 mg/kg) followed by daily doses of IG administration of EGCG were re-administered for 5 days. At the end of the experiment, blood samples were collected for analysis of biochemical parameters associated with liver. Results showed that the group of mice that were administered MESCs prior to EGCG/LPS showed lower levels of alanine amino transferase, alkaline phosphatase, and bilirubin, higher albumin/globulin ratio, and less remarkable histopathological lesions. Also, that group of mice showed less expression of oxidative stress biomarkers (oxidized low-density lipoprotein Ox.LDL and chemokine CXCL16), less expression of nuclear protein receptors (retinoic acid receptor and retinoid X receptor), and less expression of inflammatory biomarkers (tumor necrosis factor ? and transforming growth factor ?1) compared with other groups of mice that were not given MESCs. In conclusion, MESCs can ameliorate EGCG/LPS-induced hepatotoxicity in mice. PMID:25701483

  17. Ginsenoside Re ameliorates inflammation by inhibiting the binding of lipopolysaccharide to TLR4 on macrophages.

    Science.gov (United States)

    Lee, In-Ah; Hyam, Supriya R; Jang, Se-Eun; Han, Myung Joo; Kim, Dong-Hyun

    2012-09-26

    Ginseng (the root of Panax ginseng C.A. Meyer, family Araliaceae), which contains protopanaxadiol ginsenoside Rb1 and protopanaxatriol ginsenoside Re as main constituents, is frequently used to treat cancer, inflammation, and stress. In the preliminary study, protopanaxatriol ginsenoside Re inhibited NF-?B activation in lipopolysaccharide (LPS)-stimulated murine peritoneal macrophages. Therefore, we investigated its anti-inflammatory effect in peptidoglycan (PGN)-, LPS-, or tumor necrosis factor-? (TNF-?)-stimulated peritoneal macrophages and, in addition, in LPS-induced systemic inflammation and 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis in mice. Ginsenoside Re inhibited IKK-? phosphorylation and NF-?B activation, as well as the expression of proinflammatory cytokines, TNF-? and IL-1?, in LPS-stimulated peritoneal macrophages, but it did not inhibit them in TNF-?- or PG-stimulated peritoneal macrophages. Ginsenoside Re also inhibited IRAK-1 phosphorylation induced by LPS, as well as IRAK-1 and IRAK-4 degradations in LPS-stimulated peritoneal macrophages. Ginsenoside Re inhibited the binding of Alexa Fluor 488-conjugated LPS to TLR4 on peritoneal macrophages. Furthermore, ginsenoside Re inhibited the binding of LPS to TLR4 on peritoneal macrophages transiently transfected with MyD88 siRNAs. Orally administered ginsenoside Re significantly inhibited the expression of IL-1? and TNF-? on LPS-induced systemic inflammation and TNBS-induced colitis in mice. Ginsenoside Re inhibited colon shortening and myeloperoxidase activity in TNBS-treated mice. Ginsenoside Re reversed the reduced expression of tight-junction-associated proteins ZO-1, claudin-1, and occludin. Ginsenoside Re (20 mg/kg) inhibited the activation of NF-?B in TNBS-treated mice. On the basis of these findings, ginsenoside Re may ameliorate inflammation by inhibiting the binding of LPS to TLR4 on macrophages. PMID:22849695

  18. Comparison of Biological and Immunological Characterization of Lipopolysaccharides From Brucella abortus RB51 and S19

    Directory of Open Access Journals (Sweden)

    Kianmehr

    2015-11-01

    Full Text Available Background Brucella abortus RB51 is a rough stable mutant strain, which has been widely used as a live vaccine for prevention of brucellosis in cattle instead of B. abortus strain S19. B. abortus lipopolysaccharide (LPS has unique properties in comparison to other bacterial LPS. Objectives In the current study, two types of LPS, smooth (S-LPS and rough (R-LPS were purified from B. abortus S19 and RB51, respectively. The aim of this study was to evaluate biological and immunological properties of purified LPS as an immunogenical determinant. Materials and Methods Primarily, S19 and RB51 LPS were extracted and purified by two different modifications of the phenol water method. The final purity of LPS was determined by chemical analysis (2-keto-3-deoxyoctonate (KDO, glycan, phosphate and protein content and different staining methods, following sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE. C57BL/6 mice were immunized subcutaneously three times at biweekly intervals with the same amount of purified LPSs. The humoral immunity was evaluated by measuring specific IgG levels and also different cytokine levels, such as IFN-?, TNF-?, IL-4 and IL-10, were determined for assessing T-cell immune response. Results Biochemical analysis data and SDS-PAGE profile showed that the chemical nature of S19 LPS is different from RB51 LPS. Both S and R-LPS induce an immune response. T-cell immune response induced by both S and R-LPS had almost the same pattern whereas S19 LPS elicited humoral immunity, which was higher than RB51 LPS. Conclusions Purified LPS can be considered as a safe adjuvant and can be used as a component in prophylactic and therapeutic vaccines targeting infectious disease, cancer and allergies.

  19. The lipopolysaccharide core of Brucella abortus acts as a shield against innate immunity recognition.

    Science.gov (United States)

    Conde-Álvarez, Raquel; Arce-Gorvel, Vilma; Iriarte, Maite; Man?ek-Keber, Mateja; Barquero-Calvo, Elías; Palacios-Chaves, Leyre; Chacón-Díaz, Carlos; Chaves-Olarte, Esteban; Martirosyan, Anna; von Bargen, Kristine; Grilló, María-Jesús; Jerala, Roman; Brandenburg, Klaus; Llobet, Enrique; Bengoechea, José A; Moreno, Edgardo; Moriyón, Ignacio; Gorvel, Jean-Pierre

    2012-01-01

    Innate immunity recognizes bacterial molecules bearing pathogen-associated molecular patterns to launch inflammatory responses leading to the activation of adaptive immunity. However, the lipopolysaccharide (LPS) of the gram-negative bacterium Brucella lacks a marked pathogen-associated molecular pattern, and it has been postulated that this delays the development of immunity, creating a gap that is critical for the bacterium to reach the intracellular replicative niche. We found that a B. abortus mutant in the wadC gene displayed a disrupted LPS core while keeping both the LPS O-polysaccharide and lipid A. In mice, the wadC mutant induced proinflammatory responses and was attenuated. In addition, it was sensitive to killing by non-immune serum and bactericidal peptides and did not multiply in dendritic cells being targeted to lysosomal compartments. In contrast to wild type B. abortus, the wadC mutant induced dendritic cell maturation and secretion of pro-inflammatory cytokines. All these properties were reproduced by the wadC mutant purified LPS in a TLR4-dependent manner. Moreover, the core-mutated LPS displayed an increased binding to MD-2, the TLR4 co-receptor leading to subsequent increase in intracellular signaling. Here we show that Brucella escapes recognition in early stages of infection by expressing a shield against recognition by innate immunity in its LPS core and identify a novel virulence mechanism in intracellular pathogenic gram-negative bacteria. These results also encourage for an improvement in the generation of novel bacterial vaccines. PMID:22589715

  20. Lipopolysaccharide heterogeneity in the atypical group of novel emerging Brucella species.

    Science.gov (United States)

    Zygmunt, Michel S; Jacques, Isabelle; Bernardet, Nelly; Cloeckaert, Axel

    2012-09-01

    Recently, novel Brucella strains with phenotypic characteristics that were atypical for strains belonging to the genus Brucella have been reported. Phenotypically many of these strains were initially misidentified as Ochrobactrum spp. Two novel species have been described so far for these strains, i.e., B. microti and B. inopinata, and other strains genetically related to B. inopinata may constitute other novel species as well. In this study, we analyzed the lipopolysaccharides (LPS) (smooth LPS [S-LPS] and rough LPS [R-LPS]) of these atypical strains using different methods and a panel of monoclonal antibodies (MAbs) directed against several epitopes of the Brucella O-polysaccharide (O-PS) and R-LPS. Among the most striking results, Brucella sp. strain BO2, isolated from a patient with chronic destructive pneumonia, showed a completely distinct S-LPS profile in silver stain gels that looked more similar to that of enterobacterial S-LPS. This strain also failed to react with MAbs against Brucella O-PS epitopes and showed weak reactivity with anti-R-LPS MAbs. B. inopinata reference strain BO1 displayed an M-dominant S-LPS type with some heterogeneity relative to the classical M-dominant Brucella S-LPS type. Australian wild rodent strains belonging also to the B. inopinata group showed a classical A-dominant S-LPS but lacked the O-PS common (C) epitopes, as previously reported for B. suis biovar 2 strains. Interestingly, some strains also failed to react with anti-R-LPS MAbs, such as the B. microti reference strain and B. inopinata BO1, suggesting modifications in the core-lipid A moieties of these strains. These results have several implications for serological typing and serological diagnosis and underline the need for novel tools for detection and correct identification of such novel emerging Brucella spp. PMID:22761298