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Sample records for serum endotoxin lps

  1. The effect of endotoxin and anti-endotoxin serum on synovial fluid parameters in the horse

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    R.D. Gottschalk

    1998-07-01

    Full Text Available The effects of a commercially available equine hyperimmune anti-endotoxin serum on synovial fluid parameters were evaluated in an induced synovitis model in normal horses. Four groups of 3 horses each received lipopolysaccharide (LPS plus hyperimmune antiendotoxin (anti-LPS, LPS, anti-LPS, and Ringers lactate (control respectively injected into the left intercarpal joint. Synovial fluid parameters were measured at 4, 8, 24 and 72 h. It was found that anti-LPS had no attenuating effect on the LPS and that it induced a synovitis almost equivalent to that induced by LPS alone. The introduction of sterile Ringers lactate solution into the carpal joint together with repeated aseptic arthrocentesis induces a mild inflammatory response.

  2. Modeling the LPS neutralization activity of anti-endotoxins.

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    Thippakorn, Chadinee; Suksrichavalit, Thummaruk; Nantasenamat, Chanin; Tantimongcolwat, Tanawut; Isarankura-Na-Ayudhya, Chartchalerm; Naenna, Thanakorn; Prachayasittikul, Virapong

    2009-05-20

    Bacterial lipopolysaccharides (LPS), also known as endotoxins, are major structural components of the outer membrane of Gram-negative bacteria that serve as a barrier and protective shield between them and their surrounding environment. LPS is considered to be a major virulence factor as it strongly stimulates the secretion of pro-inflammatory cytokines which mediate the host immune response and culminating in septic shock. Quantitative structure-activity relationship studies of the LPS neutralization activities of anti-endotoxins were performed using charge and quantum chemical descriptors. Artificial neural network implementing the back-propagation algorithm was selected for the multivariate analysis. The predicted activities from leave-one-out cross-validation were well correlated with the experimental values as observed from the correlation coefficient and root mean square error of 0.930 and 0.162, respectively. Similarly, the external testing set also yielded good predictivity with correlation coefficient and root mean square error of 0.983 and 0.130. The model holds great potential for the rational design of novel and robust compounds with enhanced neutralization activity.

  3. Modeling the LPS Neutralization Activity of Anti-Endotoxins

    Directory of Open Access Journals (Sweden)

    Virapong Prachayasittikul

    2009-05-01

    Full Text Available Bacterial lipopolysaccharides (LPS, also known as endotoxins, are major structural components of the outer membrane of Gram-negative bacteria that serve as a barrier and protective shield between them and their surrounding environment. LPS is considered to be a major virulence factor as it strongly stimulates the secretion of pro-inflammatory cytokines which mediate the host immune response and culminating in septic shock. Quantitative structure-activity relationship studies of the LPS neutralization activities of anti-endotoxins were performed using charge and quantum chemical descriptors. Artificial neural network implementing the back-propagation algorithm was selected for the multivariate analysis. The predicted activities from leave-one-out cross-validation were well correlated with the experimental values as observed from the correlation coefficient and root mean square error of 0.930 and 0.162, respectively. Similarly, the external testing set also yielded good predictivity with correlation coefficient and root mean square error of 0.983 and 0.130. The model holds great potential for the rational design of novel and robust compounds with enhanced neutralization activity.

  4. Reduction in the bactericidal activity of selected cathelicidin peptides by bovine calf serum or exogenous endotoxin.

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    Bartlett, Karen H; McCray, Paul B; Thorne, Peter S

    2004-06-01

    Synthetic cathelicidin peptides exhibit enhanced antimicrobial action and avid binding to LPS, thereby detoxifying the action of endotoxin released from degrading bacteria. A series of cathelicidin antimicrobial peptide (CAP) and sheep myeloid antimicrobial peptide (SMAP) congeners were examined to determine whether LPS-binding could predict other beneficial characteristics of the peptides. The peptides were challenged in complex media with bovine calf serum or LPS, and their ability to kill the Gram negative pathogens Klebsiella pneumoniae (ATCC 43816) or Pseudomonas aeruginosa (PA103) was then assessed. LPS-binding efficiency was not correlated with antimicrobial activity in complex media. Additionally, LPS- and serum-binding may interfere with the antimicrobial activity of peptides in complex media. Copyright 2004 Elsevier B.V.

  5. Importance of bacterial endotoxin (LPS in endodontics A importância da endotoxina bacteriana (LPS na endodontia atual

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    Mario Roberto Leonardo

    2004-06-01

    Full Text Available New knowledge of the structure and biological activity of endotoxins (LPS has revolutionized concepts concerning their mechanisms of action and forms of inactivation. Since the 1980's, technological advances in microbiological culture and identification have shown that anaerobic microorganisms, especially Gram-negative, predominate in root canals of teeth with pulp necrosis and radiographically visible chronic periapical lesions. Gram-negative bacteria not only have different factors of virulence and generate sub-products that are toxic to apical and periapical tissues, as also contain endotoxin (LPS on their cell wall. This is especially important because endotoxin is released during multiplication or bacterial death, causing a series of biological effects that lead to an inflammatory reaction and resorption of mineralized tissues. Thus, due to the role of endotoxin in the pathogenesis of periapical lesions, we reviewed the literature concerning the biological activity of endotoxin and the relevance of its inactivation during treatment of teeth with pulp necrosis and chronic periapical lesion.O conhecimento mais aprofundado sobre a estrutura e atividade biológica das endotoxinas (LPS revolucionou os conceitos sobre seu mecanismo de ação e formas de inativação. A partir da década de 80, os avanços tecnológicos na cultura e identificação microbiológica demonstraram que, em canais radiculares de dentes portadores de necrose pulpar e lesão periapical crônica, visível radiograficamente, predominam microrganismos anaeróbios, particularmente os gram-negativos. Como se sabe, os microrganismos gram-negativos, além de possuírem diferentes fatores de virulência e gerarem produtos e sub-produtos tóxicos aos tecidos apicais e periapicais, contêm endotoxina em sua parede celular. Esse conhecimento é particularmente importante, uma vez que a endotoxina é liberada durante a multiplicação ou morte bacteriana, exercendo uma série de

  6. Translocated LPS Might Cause Endotoxin Tolerance in Circulating Monocytes of Cystic Fibrosis Patients

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    del Fresno, Carlos; Alenda, Raquel; Gómez-Piña, Vanesa; Fernández-Ruíz, Irene; Siliceo, María; Jurado, Teresa; Toledano, Victor; Arnalich, Francisco; García-Río, Francisco; López-Collazo, Eduardo

    2011-01-01

    Cystic Fibrosis (CF) is an inherited pleiotropic disease that results from abnormalities in the gene codes of a chloride channel. The lungs of CF patients are chronically infected by several pathogens but bacteraemia have rarely been reported in this pathology. Besides that, circulating monocytes in CF patients exhibit a patent Endotoxin Tolerance (ET) state since they show a significant reduction of the inflammatory response to bacterial stimulus. Despite a previous description of this phenomenon, the direct cause of ET in CF patients remains unknown. In this study we have researched the possible role of microbial/endotoxin translocation from a localized infection to the bloodstream as a potential cause of ET induction in CF patients. Plasma analysis of fourteen CF patients revealed high levels of LPS compared to healthy volunteers and patients who suffer from Chronic Obstructive Pulmonary Disease. Experiments in vitro showed that endotoxin concentrations found in plasma of CF patients were enough to induce an ET phenotype in monocytes from healthy controls. In agreement with clinical data, we failed to detect bacterial DNA in CF plasma. Our results suggest that soluble endotoxin present in bloodstream of CF patients causes endotoxin tolerance in their circulating monocytes. PMID:22216320

  7. LPS-binding protein circulates in association with apoB-containing lipoproteins and enhances endotoxin-LDL/VLDL interaction

    NARCIS (Netherlands)

    Vreugdenhil, A. C.; Snoek, A. M.; van 't Veer, C.; Greve, J. W.; Buurman, W. A.

    2001-01-01

    LPS-binding protein (LBP) and serum lipoproteins cooperate in reducing the toxic properties of LPS. In the present study, we demonstrate that LBP circulates in association with LDL and VLDL in healthy persons. ApoB was found to account at least in part for the interaction of LBP with LDL and VLDL.

  8. Inhibition of IRAK-4 activity for rescuing endotoxin LPS-induced septic mortality in mice by lonicerae flos extract

    Energy Technology Data Exchange (ETDEWEB)

    Park, Sun Hong; Roh, Eunmiri [College of Pharmacy, Chungbuk National University, Cheongju 361-763 (Korea, Republic of); Kim, Hyun Soo [Pharmaceutical R and D Center, Huons Co., Ltd., Anyang (Korea, Republic of); Baek, Seung-Il [College of Pharmacy, Chungbuk National University, Cheongju 361-763 (Korea, Republic of); Choi, Nam Song [Pharmaceutical R and D Center, Huons Co., Ltd., Anyang (Korea, Republic of); Kim, Narae; Hwang, Bang Yeon; Han, Sang-Bae [College of Pharmacy, Chungbuk National University, Cheongju 361-763 (Korea, Republic of); Kim, Youngsoo, E-mail: youngsoo@chungbuk.ac.kr [College of Pharmacy, Chungbuk National University, Cheongju 361-763 (Korea, Republic of)

    2013-12-13

    Highlights: •Lonicerae flos extract (HS-23) is a clinical candidate, Phase I for sepsis treatment. •Here, HS-23 or its major constituents rescued LPS-induced septic mortality in mice. •As a mechanism, they directly inhibited IRAK-4-catalyzed kinase activity. •Thus, they suppressed LPS-induced expression of NF-κB/AP-1-target inflammatory genes. -- Abstract: Lonicerae flos extract (HS-23) is a clinical candidate currently undergoing Phase I trial in lipopolysaccharide (LPS)-injected healthy human volunteers, but its molecular basis remains to be defined. Here, we investigated protective effects of HS-23 or its major constituents on Escherichia coli LPS-induced septic mortality in mice. Intravenous treatment with HS-23 rescued LPS-intoxicated C57BL/6J mice under septic conditions, and decreased the levels of cytokines such as tumor necrosis factor α (TNF-α), interleukin (IL)-1β and high-mobility group box-1 (HMGB-1) in the blood. Chlorogenic acid (CGA) and its isomers were assigned as major constituents of HS-23 in the protection against endotoxemia. As a molecular mechanism, HS-23 or CGA isomers inhibited endotoxin LPS-induced autophosphorylation of the IL-1 receptor-associated kinase 4 (IRAK-4) in mouse peritoneal macrophages as well as the kinase activity of IRAK-4 in cell-free reactions. HS-23 consequently suppressed downstream pathways critical for LPS-induced activation of nuclear factor (NF)-κB or activating protein 1 (AP-1) in the peritoneal macrophages. HS-23 also inhibited various toll-like receptor agonists-induced nitric oxide (NO) production, and down-regulated LPS-induced expression of NF-κB/AP-1-target inflammatory genes in the cells. Taken together, HS-23 or CGA isomers exhibited anti-inflammatory therapy against LPS-induced septic mortality in mice, at least in part, mediated through the inhibition of IRAK-4.

  9. DMPD: The role of macrophages in the hypothalamic-pituitary-adrenal activation inresponse to endotoxin (LPS). [Dynamic Macrophage Pathway CSML Database

    Lifescience Database Archive (English)

    Full Text Available 1315450 The role of macrophages in the hypothalamic-pituitary-adrenal activation in...png) (.svg) (.html) (.csml) Show The role of macrophages in the hypothalamic-pituitary-adrenal activation in...e hypothalamic-pituitary-adrenal activation inresponse to endotoxin (LPS). Authors Derijk RH, van Rooijen N,

  10. LPS infusion suppresses serum FGF21 levels in healthy adult volunteers

    DEFF Research Database (Denmark)

    Lauritzen, Esben Stistrup; Rittig, Nikolaj; Bach, Ermina

    2017-01-01

    circulating levels of FGF21 after lipopolysaccharide (LPS) infusion. DESIGN: Two randomized, single blinded, placebo-controlled crossover trials were used. SETTING: The studies were performed at a university hospital clinical research center. PATIENTS AND INTERVENTIONS: Study 1 (LPS bolus): Eight young......, healthy, lean males were investigated two times: 1) after isotonic saline injection, and 2) after LPS injection (bolus of 1 ng/kg). Each study day lasted 4 hours. Study 2 (continuous LPS infusion): Eight, healthy males were investigated two times: 1) during continuously isotonic saline infusion, and 2......) during continuously LPS infusion (0.06 ng/kg/h). Each study day lasted 4 hours. Circulating FGF21 levels were quantified every second hour by an immunoassay. RESULTS: A LPS bolus resulted in a late suppression (t = 240 minutes) of serum FGF21 (P=0.035). Continuous LPS infusion revealed no significant...

  11. Targeting naproxen coupled to human serum albumin to nonparenchymal cells reduces endotoxin-induced mortality in rats with biliary cirrhosis

    NARCIS (Netherlands)

    Albrecht, C.; Meijer, D.K F; Lebbe, C; Sägesser, H; Melgert, B.N; Poelstra, Klaas; Reichen, J

    1997-01-01

    Endotoxin is thought to play a major role in cirrhotic liver disease, Cyclo-oxygenase inhibitors were shown to be partially protective against endotoxin but cannot be used in cirrhotic patients because of renal side-effects, We argued that administration of naproxen (NAP) linked to human serum

  12. LPS infusion suppresses serum FGF21 levels in healthy adult volunteers

    DEFF Research Database (Denmark)

    Lauritzen, Esben Stistrup; Rittig, Nikolaj; Bach, Ermina

    2017-01-01

    CONTEXT: During the inflammatory acute phase response plasma glucose and serum triglycerides are increased in humans. Fibroblast growth factor (FGF) 21 has plasma glucose and lipid reducing actions, but its role in the acute inflammatory response in human is unknown. OBJECTIVE: To investigate...... circulating levels of FGF21 after lipopolysaccharide (LPS) infusion. DESIGN: Two randomized, single blinded, placebo-controlled crossover trials were used. SETTING: The studies were performed at a university hospital clinical research center. PATIENTS AND INTERVENTIONS: Study 1 (LPS bolus): Eight young...

  13. Pretreatment of lipopolysaccharide (LPS) ameliorates D-GalN/LPS induced acute liver failure through TLR4 signaling pathway.

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    Zhang, Sainan; Yang, Naibin; Ni, Shunlan; Li, Wenyuan; Xu, Lanman; Dong, Peihong; Lu, Mingqin

    2014-01-01

    Endotoxin tolerance (ET) is an important phenomenon, which affects inflammation and phagocytosis. Pretreatment with low dose of lipopolysaccharide (LPS) can protect liver injury from various hepatotoxicants such as acetaminophen and pseudomonas aeruginosa exotoxin A. The current study aimed to investigate the protecting mechanisms of endotoxin tolerance in acute liver failure induced by D-galactosamine (D-GalN)/LPS and possible role of toll-like receptors 4 (TLR4) signaling pathway in this phenomenon. Acute liver failure was induced by Injection of D-GalN/LPS. To mimic endotoxin tolerance, male Sprague-Dawley rats were treated with low dose of LPS (0.1 mg/kg once a day intraperitoneally for consecutive five days) before subsequent injection of D-GalN/LPS. Rat survival was determined by survival rate. Liver injury was confirmed by serum biochemical and liver histopathological examination. Inflammatory cytokines were determined by ELISA and nuclear factor-kappa B (NF-κB) (P65), toll-like receptors 4 (TLR4) and Interleukin-1 receptor-associated kinase-1 (IRAK-1) were measured by reverse transcriptase polymerase chain reaction and western blot respectively. Pretreatment of LPS significantly improved rat survival. Moreover, rats pretreated with LPS exhibited lower serum enzyme (ALT, AST and TBiL) level, lower production of inflammatory cytokines and more minor liver histopathological damage than rats without pretreatment of LPS. LPS pretreatment suppressed production of TLR4 and IRAK-1. LPS pretreatment also inhibited activation of hepatic NF-κB. These results indicated that endotoxin tolerance contributed to liver protection against D-GalN/LPS induced acute liver failure through down-regulation of TLR4 and NF-κB pathway.

  14. Effect of serum from various animal species on erythrocyte attachment of endotoxins and other bacterial antigens.

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    Praino, M; Neter, E

    1977-12-01

    Lipopolysaccharide O antigens (endotoxins) and other bacterial antigens readily attach to erythrocytes in vitro. This attachment is prevented by certain mammalian and avian sera. In this study, the inhibitory capacity of sera from lower animals was compared with that of higher animals for a total of 30 species. Antigens and the corresponding antisera included both crude O antigens and purified lipopolysaccharide preparations, the common enterobacterial antigen from Escherichia coli O14, the Vi antigen from Citrobacter ballerup, the polyribose-phosphate antigen from Haemophilus influenzae type b, and the crude teichoic acid antigen from Staphylococcus aureus. Antigen and serum mixtures were incubated at 37 degrees C for 30 min and used for erythrocyte modification; failure of hemagglutination by homologous bacterial antiserum provided evidence of inhibitory capacity. Sera from the classes Mammalia and Aves were very strong inhibitors; those of Reptilia and Osteichthyes were moderate in activity, displaying variation within the classes; those of Amphibia and Chondrichthyes were minimal inhibitors; and those of Merostomata, Crustacea, and Lamellibranchiata displayed questionable or no inhibitory capacity. Inhibitory sera were active with all antigens tested. The findings suggest evolution of inhibitory factors consistent with the theory of two diverging lines of animal phylogeny based on embryological criteria and closely parallel the observations of an endotoxin-altering capacity in vertebrate sera that is not found in invertebrate sera or hemolymph.

  15. Opc expression, LPS immunotype switch and pilin conversion contribute to serum resistance of unencapsulated meningococci.

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    Kerstin Hubert

    Full Text Available Neisseria meningitidis employs polysaccharides and outer membrane proteins to cope with human serum complement attack. To screen for factors influencing serum resistance, an assay was developed based on a colorimetric serum bactericidal assay. The screening used a genetically modified sequence type (ST-41/44 clonal complex (cc strain lacking LPS sialylation, polysaccharide capsule, the factor H binding protein (fHbp and MutS, a protein of the DNA repair mechanism. After killing of >99.9% of the bacterial cells by serum treatment, the colorimetric assay was used to screen 1000 colonies, of which 35 showed enhanced serum resistance. Three mutant classes were identified. In the first class of mutants, enhanced expression of Opc was identified. Opc expression was associated with vitronectin binding and reduced membrane attack complex deposition confirming recent observations. Lipopolysaccharide (LPS immunotype switch from immunotype L3 to L8/L1 by lgtA and lgtC phase variation represented the second class. Isogenic mutant analysis demonstrated that in ST-41/44 cc strains the L8/L1 immunotype was more serum resistant than the L3 immunotype. Consecutive analysis revealed that the immunotypes L8 and L1 were frequently observed in ST-41/44 cc isolates from both carriage and disease. Immunotype switch to L8/L1 is therefore suggested to contribute to the adaptive capacity of this meningococcal lineage. The third mutant class displayed a pilE allelic exchange associated with enhanced autoaggregation. The mutation of the C terminal hypervariable region D of PilE included a residue previously associated with increased pilus bundle formation. We suggest that autoaggregation reduced the surface area accessible to serum complement and protected from killing. The study highlights the ability of meningococci to adapt to environmental stress by phase variation and intrachromosomal recombination affecting subcapsular antigens.

  16. Intralaboratory validation of kinetic chromogenic Limulus amebocyte lysate assay for bacterial endotoxin determination in anti-bothropic serum.

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    Fingola, Fernando F; Albertino, Sheila R G; Abrantes, Shirley de M P; Zamith, Helena P S

    2013-11-01

    Over the years, substituting in vitro biological methods for in vivo tests has posed an ever increasing challenge for researchers, including those who study the applications for snake antivenom. In the quality control of antivenons, the only official test recommended by pharmacopoeias for detecting pyrogenicity is the rabbit pyrogen test. In the present study, we propose intralaboratory validation of a method to replace the rabbit pyrogen test: in vitro determination of bacterial endotoxin in anti-bothropic serum (ABS) with quantitative kinetic chromogenic limulus amebocyte lysate (LAL) assay. The kinetic chromogenic LAL assay is specific to the detection of gram-negative bacterial endotoxin. The validation of the test involved the determination of performance parameters required by the Agência Nacional de Vigilância Sanitária Brasileira (ANVISA, the Brazilian National Health Surveillance Agency), the United States Food and Drug Administration (FDA) and the United States Pharmacopeia (USP) 35. In all experiments, the correlation coefficient of the curve obtained with the control standard endotoxin (CSE; Escherichia coli 055:B5 strain, range, 0.005-50 EU/mL) was between -0.998 and -1.000; and the recovery of endotoxin added to the sample of ABS (0.5 EU/mL) at the working dilution (1:10) followed the recuperation criteria (i.e., 50-200%). We performed six determinations, in each of which the coefficient of variation for the intermediate precision was between 5.6% and 13.8% (below the 15% threshold) and the accuracy was between 90.7% and 114.3% (within the acceptable range of 80-120%). The endotoxin concentration limit for the ABS was determined to be ≤ 2.9 EU/mL. The intralaboratory validation of the methodology was considered to have been successful because it met the criteria for all of the performance parameters. Copyright © 2013 Elsevier B.V. All rights reserved.

  17. Dehydrated citrus pulp alters feedlot performance of crossbred heifers during the receiving period and modulates serum metabolite concentrations before and after an endotoxin challenge.

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    Cribbs, J T; Bernhard, B C; Young, T R; Jennings, M A; Burdick Sanchez, N C; Carroll, J A; Callaway, T R; Schmidt, T B; Johnson, B J; Rathmann, R J

    2015-12-01

    English × Continental heifers ( = 180) were sourced in 2 loads (219.3 ± 16.0 and 221.4 ± 16.4 kg, respectively) from commercial auction barns to study the effects of feeding dehydrated citrus pulp (DCP) on feedlot performance of newly received heifers. A completely randomized block design was used with BW nested within arrival load and blocked by BW into 3 dietary treatments (36 pens, 5 heifers/pen, 12 blocks, 3 pens/block, and 12 pens/treatment). Treatment diets contained 1) 0% DCP (control diet [CON]), 2) 10% DCP, or 3) 20% DCP on a DM basis. Diets containing DCP were exchanged with steam-flaked corn on a 1:1 basis. Cattle were fed a 63, 73, and 83% concentrate diet from d 0 to 28, d 28 to 42, and d 42 to 56, respectively. Over the 56-d trial period, as the amount of dietary DCP increased, DMI decreased ( = 0.01), ADG decreased ( < 0.01), and G:F decreased ( = 0.02). From d 0 to 28, there was no difference in the observed minus the predicted NEg of the diet ( = 0.73); from d 28 to 42, there was a linear increase in NEg favoring DCP treatments ( < 0.01); and from d 42 to 56, there was a linear decrease in NEg against the DCP treatments ( < 0.01). At the conclusion of the trial, a subset of heifers ( = 22; 307.89 ± 3.32 kg on d 63) were used to evaluate blood metabolite concentrations before and after a lipopolysaccharide (LPS) challenge. On d 63, heifers were fitted with jugular catheters and moved into individual stalls. On d 64, heifers were intravenously challenged with LPS (0.5 μg/kg BW), and blood samples were collected every 0.5 h from -2 to 8 h and at 24 h relative to the LPS challenge (0 h). Serum glucose, serum urea nitrogen (SUN), and NEFA concentrations were determined. Cattle lost less weight at both 24 and 72 h after the LPS challenge with increasing DCP percentage ( < 0.01). Glucose ( = 0.12) and NEFA ( = 0.13) concentrations did not differ before the LPS challenge; however, there was a treatment effect for SUN, with elevated concentrations of

  18. The crosstalk between Nrf2 and AMPK signal pathways is important for the anti-inflammatory effect of berberine in LPS-stimulated macrophages and endotoxin-shocked mice.

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    Mo, Chunfen; Wang, Ling; Zhang, Jie; Numazawa, Satoshi; Tang, Hong; Tang, Xiaoqiang; Han, Xiaojuan; Li, Junhong; Yang, Ming; Wang, Zhe; Wei, Dandan; Xiao, Hengyi

    2014-02-01

    The response of AMP-activated protein kinase (AMPK) to oxidative stress has been recently reported but the downstream signals of this response are largely unknown. Meanwhile, the upstream events for the activation of nuclear factor erythroid-2-related factor-2 (Nrf2), a critical transcriptional activator for antioxidative responses, remain unclear. In the present study, we investigated the relationship between AMPK and Nrf2 signal pathways in lipopolysaccharide (LPS)-triggered inflammatory system, in which berberine (BBR), a known AMPK activator, was used for inflammation suppression. In inflammatory macrophages, BBR attenuated LPS-induced expression of inflammatory genes (inducible nitric oxide synthase [iNOS], cyclooxygenase-2 [COX2], interleukin [IL]-6), and the generation of nitric oxide and reactive oxygen species, but increased the transcription of Nrf2-targeted antioxidative genes (NADPH quinone oxidoreductase-1 [NQO-1], heme oxygenase-1 [HO-1]), as well as the nuclear localization and phosphorylation of Nrf2 protein. Importantly, we found BBR-induced activation of Nrf2 is AMPK-dependent, as either pharmacologically or genetically inactivating AMPK blocked the activation of Nrf2. Consistent with in vitro experiments, BBR down-regulated the expression of proinflammatory genes but upregulated those of Nrf2-targeted genes in lungs of LPS-injected mice, and these effects were attenuated in Nrf2-deficient mice. Moreover, the effect of BBR on survival time extension and plasma redox regulation in endotoxin-shocked mice was largely weakened when Nrf2-depleted. Our results demonstrate convergence between AMPK and Nrf2 pathways and this intersection is essential for anti-inflammatory effect of BBR in LPS-stimulated macrophages and endotoxin-shocked mice. Uncovering this intersection is significant for understanding the relationship between energy homeostasis and antioxidative responses and may be beneficial for developing new therapeutic strategies against

  19. Serum Endotoxins and Flagellin and Risk of Colorectal Cancer in the European Prospective Investigation into Cancer and Nutrition (EPIC) Cohort

    Science.gov (United States)

    Kong, So Yeon; Tran, Hao Quang; Gewirtz, Andrew T.; McKeown-Eyssen, Gail; Fedirko, Veronika; Romieu, Isabelle; Tjønneland, Anne; Olsen, Anja; Overvad, Kim; Boutron-Ruault, Marie-Christine; Bastide, Nadia; Affret, Aurélie; Kühn, Tilman; Kaaks, Rudolf; Boeing, Heiner; Aleksandrova, Krasimira; Trichopoulou, Antonia; Kritikou, Maria; Vasilopoulou, Effie; Palli, Domenico; Krogh, Vittorio; Mattiello, Amalia; Tumino, Rosario; Naccarati, Alessio; Bueno-de-Mesquita, H.Bas; Peeters, Petra H.; Weiderpass, Elisabete; Quirós, J. Ramón; Sala, Núria; Sánchez, María-José; Huerta Castaño, José María; Barricarte, Aurelio; Dorronsoro, Miren; Werner, Mårten; Wareham, Nicholas J.; Khaw, Kay-Tee; Bradbury, Kathryn E.; Freisling, Heinz; Stavropoulou, Faidra; Ferrari, Pietro; Gunter, Marc J.; Cross, Amanda J.; Riboli, Elio; Bruce, W. Robert

    2017-01-01

    Background Chronic inflammation and oxidative stress are thought to be involved in colorectal cancer (CRC) development. These processes may be contributed to by leakage of bacterial products, such as lipopolysaccharide (LPS) and flagellin, across the gut barrier. The objective of this study, nested within a prospective cohort, was to examine associations between circulating LPS and flagellin serum antibody levels and CRC risk. Methods 1,065 incident CRC cases (colon n=667; rectal n=398) were matched (1:1) to control subjects. Serum flagellin- and LPS-specific IgA and IgG levels were quantitated by ELISA. Multivariable conditional logistic regression models were used to calculate odds ratios (OR) and 95% confidence intervals (CI), adjusting for multiple relevant confouding factors. Results Overall, elevated anti-LPS and anti-flagellin biomarker levels were not associated with CRC risk. After testing potential interactions by various factors relevant for CRC risk and anti-LPS and anti-flagellin, sex was identified as a statistically significant interaction factor (pinteraction < 0.05 for all the biomarkers). Analyses stratified by sex showed a statistically significant positive CRC risk association for men (fully-adjusted OR for highest vs. lowest quartile for total anti-LPS+flagellin = 1.66; 95% CI, 1.10-2.51; ptrend = 0.049) while a borderline statistically significant inverse association was observed for women (fully-adjusted OR= 0.70; 95%CI, 0.47-1.02; ptrend = 0.18). Conclusion In this prospective study on European populations, we found bacterial exposure levels to be positively associated to CRC risk among men while in women, a possible inverse association may exist. Impact Further studies are warranted to better clarify these preliminary observations. PMID:26823475

  20. Serum Endotoxins and Flagellin and Risk of Colorectal Cancer in the European Prospective Investigation into Cancer and Nutrition (EPIC) Cohort.

    Science.gov (United States)

    Kong, So Yeon; Tran, Hao Quang; Gewirtz, Andrew T; McKeown-Eyssen, Gail; Fedirko, Veronika; Romieu, Isabelle; Tjønneland, Anne; Olsen, Anja; Overvad, Kim; Boutron-Ruault, Marie-Christine; Bastide, Nadia; Affret, Aurélie; Kühn, Tilman; Kaaks, Rudolf; Boeing, Heiner; Aleksandrova, Krasimira; Trichopoulou, Antonia; Kritikou, Maria; Vasilopoulou, Effie; Palli, Domenico; Krogh, Vittorio; Mattiello, Amalia; Tumino, Rosario; Naccarati, Alessio; Bueno-de-Mesquita, H B; Peeters, Petra H; Weiderpass, Elisabete; Quirós, J Ramón; Sala, Núria; Sánchez, María-José; Castaño, José María Huerta; Barricarte, Aurelio; Dorronsoro, Miren; Werner, Mårten; Wareham, Nicholas J; Khaw, Kay-Tee; Bradbury, Kathryn E; Freisling, Heinz; Stavropoulou, Faidra; Ferrari, Pietro; Gunter, Marc J; Cross, Amanda J; Riboli, Elio; Bruce, W Robert; Jenab, Mazda

    2016-02-01

    Chronic inflammation and oxidative stress are thought to be involved in colorectal cancer development. These processes may contribute to leakage of bacterial products, such as lipopolysaccharide (LPS) and flagellin, across the gut barrier. The objective of this study, nested within a prospective cohort, was to examine associations between circulating LPS and flagellin serum antibody levels and colorectal cancer risk. A total of 1,065 incident colorectal cancer cases (colon, n = 667; rectal, n = 398) were matched (1:1) to control subjects. Serum flagellin- and LPS-specific IgA and IgG levels were quantitated by ELISA. Multivariable conditional logistic regression models were used to calculate ORs and 95% confidence intervals (CI), adjusting for multiple relevant confouding factors. Overall, elevated anti-LPS and anti-flagellin biomarker levels were not associated with colorectal cancer risk. After testing potential interactions by various factors relevant for colorectal cancer risk and anti-LPS and anti-flagellin, sex was identified as a statistically significant interaction factor (Pinteraction < 0.05 for all the biomarkers). Analyses stratified by sex showed a statistically significant positive colorectal cancer risk association for men (fully-adjusted OR for highest vs. lowest quartile for total anti-LPS + flagellin, 1.66; 95% CI, 1.10-2.51; Ptrend, 0.049), whereas a borderline statistically significant inverse association was observed for women (fully-adjusted OR, 0.70; 95% CI, 0.47-1.02; Ptrend, 0.18). In this prospective study on European populations, we found bacterial exposure levels to be positively associated to colorectal cancer risk among men, whereas in women, a possible inverse association may exist. Further studies are warranted to better clarify these preliminary observations. ©2016 American Association for Cancer Research.

  1. Anti-RAGE antibody selectively blocks acute systemic inflammatory responses to LPS in serum, liver, CSF and striatum.

    Science.gov (United States)

    Gasparotto, Juciano; Ribeiro, Camila Tiefensee; Bortolin, Rafael Calixto; Somensi, Nauana; Fernandes, Henrique Schaan; Teixeira, Alexsander Alves; Guasselli, Marcelo Otavio Rodrigues; Agani, Crepin Aziz Jose O; Souza, Natália Cabral; Grings, Mateus; Leipnitz, Guilhian; Gomes, Henrique Mautone; de Bittencourt Pasquali, Matheus Augusto; Dunkley, Peter R; Dickson, Phillip W; Moreira, José Claudio Fonseca; Gelain, Daniel Pens

    2017-05-01

    Systemic inflammation induces transient or permanent dysfunction in the brain by exposing it to soluble inflammatory mediators. The receptor for advanced glycation endproducts (RAGE) binds to distinct ligands mediating and increasing inflammatory processes. In this study we used an LPS-induced systemic inflammation model in rats to investigate the effect of blocking RAGE in serum, liver, cerebrospinal fluid (CSF) and brain (striatum, prefrontal cortex, ventral tegmental area and substantia nigra). Intraperitoneal injection of RAGE antibody (50μg/kg) was followed after 1h by a single LPS (5mg/kg) intraperitoneal injection. Twenty-four hours later, tissues were isolated for analysis. RAGE antibody reduced LPS-induced inflammatory effects in both serum and liver; the levels of proinflammatory cytokines (TNF-α, IL-1β) were decreased and the phosphorylation/activation of RAGE downstream targets (ERK1/2, IκB and p65) in liver were significantly attenuated. RAGE antibody prevented LPS-induced effects on TNF-α and IL-1β in CSF. In striatum, RAGE antibody inhibited increases in IL-1β, Iba-1, GFAP, phospho-ERK1/2 and phospho-tau (ser202), as well as the decrease in synaptophysin levels. These effects were caused by systemic RAGE inhibition, as RAGE antibody did not cross the blood-brain barrier. RAGE antibody also prevented striatal lipoperoxidation and activation of mitochondrial complex II. In conclusion, blockade of RAGE is able to inhibit inflammatory responses induced by LPS in serum, liver, CSF and brain. Copyright © 2017 Elsevier Inc. All rights reserved.

  2. The effect of probiotics on serum levels of cytokine and endotoxin in peritoneal dialysis patients: a randomised, double-blind, placebo-controlled trial.

    Science.gov (United States)

    Wang, I-K; Wu, Y-Y; Yang, Y-F; Ting, I-W; Lin, C-C; Yen, T-H; Chen, J-H; Wang, C-H; Huang, C-C; Lin, H-C

    2015-01-01

    Inflammatory markers such as interleukin (IL)-6 and tumour necrosis factor-alpha (TNF-α) are elevated in dialysis patients and can predict cardiovascular events and all-cause mortality. Endotoxin is an important source and also another marker of inflammation in patients with chronic kidney disease. The aim of this study was to evaluate the impact of oral probiotics on serum levels of endotoxemia and cytokines in peritoneal dialysis (PD) patients. The decline of residual renal function, peritonitis episodes, and cardiovascular events were also recorded. From July 2011 to June 2012, a randomised, double-blind, placebo-controlled trial was conducted in PD patients. The intervention group received one capsule of probiotics containing 10(9) cfu Bifobacterium bifidum A218, 10(9) cfu Bifidobacterium catenulatum A302, 10(9) cfu Bifidobacterium longum A101, and 10(9) cfu Lactobacillus plantarum A87 daily for six months, while the placebo group received similar capsules containing maltodextrin for the same duration. Levels of serum TNF-α, interferon gamma, IL-5, IL-6, IL-10, IL-17, and endotoxin were measured before and six months after intervention. 39 patients completed the study (21 in the probiotics group and 18 in the placebo group). In patients receiving probiotics, levels of serum TNF-α, IL-5, IL-6, and endotoxin significantly decreased after six months of treatment, while levels of serum IL-10 significantly increased. In contrast, there were no significant changes in levels of serum cytokines and endotoxin in the placebo group after six months. In addition, the residual renal function was preserved in patients receiving probiotics. In conclusion, probiotics could significantly reduce the serum levels of endotoxin, pro-inflammatory cytokines (TNF-α and IL-6), IL-5, increase the serum levels of anti-inflammatory cytokine (IL-10), and preserve residual renal function in PD patients.

  3. Thermal and cytokine responses to endotoxin challenge during early life.

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    McDonald, Fiona B; Khawaja, Ahmad M; Imran, Ahmad A; Ellis, Margot E; Chandrasekharan, Kumaran; Hasan, Shabih U

    2017-12-01

    Sudden infant death syndrome (SIDS) remains the leading cause of infant mortality beyond the neonatal period. An increase in body temperature as a result of high environmental temperature, overwrapping of infants, and (or) infection are associated with SIDS. Endotoxins such as lipopolysaccharide (LPS) and heat stress may perturb cardiorespiratory function and thermoregulation. Although LPS-mediated body temperature and cytokine responses are well documented in older animals, the capacity of LPS to induce fever and cytokine response in young rats remains unclear. Therefore, we sought to investigate the acute effects of LPS on body temperature and cytokine concentrations in rat pups. Postnatal day 7 rat pups were divided into 3 groups: Group 1, rats were administered LPS intraperitoneally (200 μg/kg); Group 2, rats received saline at volume equal to that administered in the LPS group; Group 3, rats received no treatment. Pups were placed in custom-made chambers maintained at ambient temperature of 33 °C. Body surface temperature was continuously monitored for 4 h. Thereafter, the rats were euthanized and serum was collected for cytokine analysis. We demonstrate that LPS treatment increased MIP-1α, IL-10, MCP-1, IP-10, fractalkine, and TNF-α with no concurrent rise in body surface temperature. Although neonatal rats produced an array of cytokines in response to LPS, there was no evidence of fever.

  4. Artesunate Reduces Serum Lipopolysaccharide in Cecal Ligation/Puncture Mice via Enhanced LPS Internalization by Macrophages through Increased mRNA Expression of Scavenger Receptors

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    Bin Li

    2014-01-01

    Full Text Available Innate immunity is the first line of defense in human beings against pathogen infection; monocytes/macrophages are the primary cells of the innate immune system. Recently, macrophages/monocytes have been discovered to participate in LPS clearance, and the clearance efficiency determines the magnitude of the inflammatory response and subsequent organ injury. Previously, we reported that artesunate (AS protected sepsis mice against heat-killed E. coli challenge. Herein, we further confirmed that AS protected cecal ligation/puncture (CLP sepsis mice. Its protection on sepsis mice was related to not only reduction of pro-inflammatory cytokines and serum LPS levels but also improvement of liver function. Based on the fact that AS did not directly bind and neutralize LPS, we hypothesized that the reduction of serum LPS level might be related to enhancement of LPS internalization and subsequent detoxification. Our results showed that AS increased FITC-LPS internalization by peritoneal macrophage and liver Kupffer cell, but enhancement of LPS internalization by AS was not related to the clathrin-dependent pathway. However, AS induced mRNA expression of important scavenger receptors (SRs; SR-A and MARCO mRNA expression was upregulated, suggesting that AS enhancement of LPS internalization and inhibition of pro-inflammatory cytokines was related to changes in mRNA expression of SRs.

  5. Duration of in vivo endotoxin tolerance in horses.

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    Holcombe, Susan J; Jacobs, Carrie C; Cook, Vanessa L; Gandy, Jeffery C; Hauptman, Joseph G; Sordillo, Lorraine M

    2016-05-01

    Endotoxemia models are used to study mechanisms and treatments of early sepsis. Repeated endotoxin exposures induce periods of endotoxin tolerance, characterized by diminished proinflammatory responses to lipopolysaccharide (LPS) and modulated production of proinflammatory cytokines. Repeated measure designs using equine endotoxemia models are rarely performed, despite the advantages associated with reduced variability, because the altered responsiveness would confound study results and because the duration of equine endotoxin tolerance is unknown. We determined the interval of endotoxin tolerance, in vivo, in horses based on physical, clinicopathologic, and proinflammatory gene expression responses to repeated endotoxin exposures. Six horses received 30 ng/kg LPS in saline infused over 30 min. Behavior pain scores, physical examination parameters, and blood for complete blood count and proinflammatory gene expression were obtained at predetermined intervals for 24h. Horses received a total of 3 endotoxin exposures. The first exposure was LPS 1, followed 7 days later by LPS 7 or 14-21 days later by LPS 14-21. Lipopolysaccharide exposures were allocated in a randomized, crossover design. Lipopolysaccharide produced clinical and clinicopathologic signs of endotoxemia and increased expression of tumor necrosis factor alpha (TNFα), interleukin (IL)-6 and IL-8, PHorses exhibited evidence of endotoxin tolerance following LPS 7 but not following LPS 14-21. Horses had significantly lower pain scores, heart rates, respiratory rates and duration of fever, after LPS 7 compared to LPS 1 and LPS 14-21, Phorses after LPS 7, P=0.05. Clinical parameters and TNFα gene expression were similar or slightly increased in horses following LPS 14-21 compared to measurements made in horses following LPS 1, suggesting that endotoxin tolerance had subsided. A minimum of 3 weeks between experiments is warranted if repeated measures designs are used to assess in vivo response to endotoxin in

  6. Inhibition of NLRP3 inflammasome: a new protective mechanism of cinnamaldehyde in endotoxin poisoning of mice.

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    Xu, Feng; Wang, Feng; Wen, Taoqun; Sang, Wentao; Wang, Dejian; Zeng, Nan

    2017-10-01

    Cinnamaldehyde (CA) has a protective effect in endotoxin poisoning of mice, but there is no direct evidence for the protective effect of CA through inhibition of NLRP3 inflammasome activation in endotoxin poisoning of mice. We aimed to investigate the protective mechanism of CA in endotoxin poisoned mice through NLRP3 inflammasome. First, we evaluated the anti-inflammatory effect of CA in phorbol-12-myristate acetate-differentiated THP-1 cells through the NLRP3 inflammasome. Second, in a mouse model of lipopolysaccharide (LPS)-induced endotoxin poisoning, CA was administrated for 5 d (once a day) before the 15 mg/kg LPS challenge. Then, the levels of IL-1β in serum were measured, and the effect of CA on the NLRP3 inflammasome activation and the expression of cathepsin B and P2X7R proteins in lung were explored. In vitro, CA decreased the levels of p20, pro-IL-1β and IL-1β in cell culture supernatants, as well as the expression of NLRP3 and IL-1β mRNA in cells. In vivo, CA decreased IL-1β production in serum. Furthermore, CA suppressed LPS-induced NLRP3, p20, Pro-IL-1β, P2X7 receptor (P2X7R) and cathepsin B protein expression in lung, as well as the expression of NLRP3 and IL-1β mRNA. CA has a protective effect in the endotoxin poisoned mice through the inhibition of NLRP3 inflammasome activation. Furthermore, CA suppresses the NLRP3 inflammasome activation by inhibiting the expression of cathepsin B and P2X7R protein expression. CA can be considered as a potential therapeutic candidate for diseases involved in endotoxin poisoning such as sepsis.

  7. Common epitopes in LPS of different Enterobacteriaceae are associated with an immune response against Escherichia coli O157 in bovine serum samples.

    Science.gov (United States)

    Navarro, Armando; Eslava, Carlos; García de la Torre, Guadalupe; León, Luis Antonio; Licona, Delia; León, Lemuel; Zarco, Luis Alberto; Cravioto, Alejandro

    2007-11-01

    Epidemiological studies in both humans and animals conducted in Mexico have shown that the isolation frequency of Escherichia coli O157 : H7 is low. In a previous study, IgG antibodies against E. coli O157, O7 and O116 LPS were found in serum samples from children and adults with no previous history of E. coli O157 : H7 infection. The present study was designed to determine whether a similar immune response against E. coli O157 : H7 and other antigenically related bacteria was present in bovine serum samples. A total of 310 serum samples from different herds in Mexico was analysed by microagglutination assays against different enterobacterial antigens, including E. coli O157. Microagglutination assays were positive against E. coli O7 (55 %), O116 (76 %) and O157 (36 %), Escherichia hermannii (15 %), Salmonella enterica serotype Urbana (14 %) and Salmonella enterica subsp. arizonae (40 %). These results were confirmed using a specific ELISA with purified LPS. A positive reaction was observed against the LPS of E. coli O7 (29 %), O116 (12 %) and O157 (22 %), E. hermannii (4 %), Salmonella Urbana (13 %) and S. enterica subsp. arizonae (12 %). Serum absorption studies of positive serum samples indicated the existence of at least three common epitopes shared by the LPS of E. coli O7, O116 and O157, and two others between E. coli O157 and Salmonella Urbana and S. enterica subsp. arizonae. A bactericidal assay against E. coli O157 : H7 using 31 bovine serum samples was performed, and 22 (71 %) of these serum samples gave positive results. The data demonstrated that bovine serum showed a response against different enterobacteria, including E. coli O157, and that this response could be due to the presence of shared epitopes in the LPS of these organisms.

  8. Endotoxin induced peritonitis elicits monocyte immigration into the lung: implications on alveolar space inflammatory responsiveness

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    Welte Tobias

    2006-02-01

    Full Text Available Abstract Background Acute peritonitis developing in response to gram-negative bacterial infection is known to act as a trigger for the development of acute lung injury which is often complicated by the development of nosocomial pneumonia. We hypothesized that endotoxin-induced peritonitis provokes recruitment of monocytes into the lungs, which amplifies lung inflammatory responses to a second hit intra-alveolar challenge with endotoxin. Methods Serum and lavage cytokines as well as bronchoalveolar lavage fluid cells were analyzed at different time points after intraperitoneal or intratracheal application of LPS. Results We observed that mice challenged with intraperitoneal endotoxin developed rapidly increasing serum and bronchoalveolar lavage fluid (BALF cytokine and chemokine levels (TNFα, MIP-2, CCL2 and a nearly two-fold expansion of the alveolar macrophage population by 96 h, but this was not associated with the development of neutrophilic alveolitis. In contrast, expansion of the alveolar macrophage pool was not observed in CCR2-deficient mice and in wild-type mice systemically pretreated with the anti-CD18 antibody GAME-46. An intentional two-fold expansion of alveolar macrophage numbers by intratracheal CCL2 following intraperitoneal endotoxin did not exacerbate the development of acute lung inflammation in response to intratracheal endotoxin compared to mice challenged only with intratracheal endotoxin. Conclusion These data, taken together, show that intraperitoneal endotoxin triggers a CCR2-dependent de novo recruitment of monocytes into the lungs of mice but this does not result in an accentuation of neutrophilic lung inflammation. This finding represents a previously unrecognized novel inflammatory component of lung inflammation that results from endotoxin-induced peritonitis.

  9. Evidence for endotoxin contamination in plastic Na+-heparin blood collection tube lots

    National Research Council Canada - National Science Library

    Newhall, Kathryn J; Diemer, Geoffrey S; Leshinsky, Natalia; Kerkof, Keith; Chute, Hilary T; Russell, Chris B; Rees, William; Welcher, Andrew A; Patterson, Scott D; Means, Gary D

    2010-01-01

    .... Because bacterial lipopolysaccharide (LPS) (endotoxin) contamination is known to cause spontaneous cytokine production by monocytes, contamination of blood collection tubes may interfere with biomarker assay results...

  10. Evidence for endotoxin contamination in plastic [Na.sup.+]-heparin blood collection tube lots

    National Research Council Canada - National Science Library

    Rees, William; Patterson, Scott D; Chute, Hilary T; Welcher, Andrew A; Russell, Chris B; Means, Gary D; Kerkof, Keith; Diemer, Geoffrey S; Newhall, Kathryn J; Leshinsky, Natalia

    2010-01-01

    .... Because bacterial lipopolysaccharide (LPS) (endotoxin) contamination is known to cause spontaneous cytokine production by monocytes, contamination of blood collection tubes may interfere with biomarker assay results. METHODS...

  11. Microcirculatory alterations of hepatic and mesenteric microcirculation in endotoxin tolerance.

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    Lendemans, Sven; Peszko, Adam; Oberbeck, Reiner; Schmitz, Daniel; Husain, Baher; Burkhard, Marcus; Schade, Fritz Ulrich; Flohe, Sascha

    2008-02-01

    LPSs getting access to the circulation of mammalian organisms cause typical systemic inflammatory reactions with symptoms characteristic for acute sepsis. One possibility to attenuate LPS effects is to expose a host to a challenge with low LPS doses, which results in the establishment of "endotoxin tolerance" (ET). Because the microcirculation is of particular importance in LPS action, it seemed of interest to analyze leukocyte-endothelial interactions in the mesentery and liver once endotoxin tolerance has been established and are challenged with LPS. The mesenteric and hepatic microcirculation was investigated by intravital microscopy. After induction of ET LPS, shock was induced by i.v. injection of LPS, and microcirculation of the mesentery and liver was examined. Endotoxin tolerance resulted in reduced ex vivo TNF-alpha synthesis of whole blood. In vivo LPS caused no increase of body temperature. In sinusoids, LPS challenge increased adherence of leukocytes in naive rats, which was almost completely prevented by ET induction. In contrast, in postsinusoidal venules, leukocyte adherence was more intense after ET induction and subsequent to LPS application. Similarly, in postcapillary mesenteric venules, increased adherence of leukocytes after LPS challenge in the ET group was observed. After LPS injection, the endothelial barrier was more disturbed in the nontolerant group when compared with the ET group. Soluble L-selectin and intercellular adhesion molecule were elevated in both ET and untreated rats. Endotoxin tolerance influences leukocyte-endothelial interaction differentially depending on organ and vessel area.

  12. Aspirin-triggered resolvin D1 down-regulates inflammatory responses and protects against endotoxin-induced acute kidney injury

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    Chen, Jiao [Center for Research on Environmental Disease, University of Kentucky, Lexington, KY 40536 (United States); Shetty, Sreerama [Center for Biomedical Research, University of Texas Health Science Center at Tyler, Tyler, TX 75708 (United States); Zhang, Ping [State Key Laboratory of Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu 610041 (China); Gao, Rong; Hu, Yuxin [Center for Research on Environmental Disease, University of Kentucky, Lexington, KY 40536 (United States); Wang, Shuxia [Graduate Center for Nutritional Sciences, College of Medicine, University of Kentucky, Lexington, KY 40536 (United States); Li, Zhenyu [Division of Cardiovascular Medicine, University of Kentucky, Lexington, KY 40536 (United States); Fu, Jian, E-mail: jian.fu@uky.edu [Center for Research on Environmental Disease, University of Kentucky, Lexington, KY 40536 (United States); Graduate Center for Toxicology, University of Kentucky, Lexington, KY 40536 (United States)

    2014-06-01

    The presence of endotoxin in blood can lead to acute kidney injury (AKI) and septic shock. Resolvins, the endogenous lipid mediators derived from docosahexaenoic acid, have been reported to exhibit potent anti-inflammatory action. Using a mouse model of lipopolysaccharide (LPS)-induced AKI, we investigated the effects of aspirin-triggered resolvin D1 (AT-RvD1) on inflammatory kidney injury. Administration of AT-RvD1 1 h after LPS challenge protected the mice from kidney injury as indicated by the measurements of blood urea nitrogen, serum creatinine, and morphological alterations associated with tubular damage. The protective effects were evidenced by decreased neutrophil infiltration in the kidney indicating reduction in inflammation. AT-RvD1 treatment restored kidney cell junction protein claudin-4 expression, which was otherwise reduced after LPS challenge. AT-RvD1 treatment inhibited endotoxin-induced NF-κB activation and suppressed LPS-induced ICAM-1 and VCAM-1 expression in the kidney. Moreover, AT-RvD1 treatment markedly decreased LPS-induced IL-6 level in the kidney and blocked IL-6-mediated signaling including STAT3 and ERK phosphorylation. Our findings demonstrate that AT-RvD1 is a potent anti-inflammatory mediator in LPS-induced kidney injury, and AT-RvD1 has therapeutic potential against AKI during endotoxemia.

  13. Increase in hypothalamic AMPK phosphorylation induced by prolonged exposure to LPS involves ghrelin and CB1R signaling.

    Science.gov (United States)

    Rivas, Priscila M S; Vechiato, Fernanda M V; Borges, Beatriz C; Rorato, Rodrigo; Antunes-Rodrigues, Jose; Elias, Lucila L K

    2017-07-01

    Acute administration of lipopolysaccharide (LPS) from Gram-negative bacteria induces hypophagia. However, the repeated administration of LPS leads to desensitization of hypophagia, which is associated with increased hypothalamic p-AMPK expression. Because ghrelin and endocannabinoids modulate AMPK activity in the hypothalamus, we hypothesized that these neuromodulators play a role in the reversal of tolerance to hypophagia in rats under long-term exposure to LPS. Male Wistar rats were treated with single (1 LPS, 100μg/kg body weight, ip) or repeated injections of LPS over 6days (6 LPS). Food intake was reduced in the 1 LPS, but not in the 6 LPS group. 6 LPS rats showed an increased serum concentration of acylated ghrelin and reduced ghrelin receptor mRNA expression in the hypothalamus. Ghrelin injection (40μg/kg body weight, ip) increased food intake, body weight gain, p-AMPK hypothalamic expression, neuropeptide Y (NPY) and Agouti related peptide (AgRP) mRNA expression in control animals (Saline). However, in 6 LPS rats, ghrelin did not alter these parameters. Central administration of a CB1R antagonist (AM251, 200ng/μl in 5μl/rat) induced hypophagia in 6 LPS animals, suggesting that the endocannabinoid system contributes to preserved food intake during LPS tolerance. In the presence of AM251, the ability of ghrelin to phosphorylate AMPK in the hypothalamus of 6 LPS group was restored, but not its orexigenic effect. Our data highlight that the orexigenic effects of ghrelin require CB1R signaling downstream of AMPK activation. Moreover, CB1R-mediated pathways contribute to the absence of hypophagia during repeated exposure to endotoxin. Copyright © 2017 Elsevier Inc. All rights reserved.

  14. Gene-environment interactions between CD14 C-260T and endotoxin exposure on Foxp3+ and Foxp3- CD4+ lymphocyte numbers and total serum IgE levels in early childhood.

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    Williams, L Keoki; Oliver, Jennifer; Peterson, Edward L; Bobbitt, Kevin R; McCabe, Michael J; Smolarek, Derek; Havstad, Suzanne L; Wegienka, Ganesa; Burchard, Esteban G; Ownby, Dennis R; Johnson, Christine C

    2008-02-01

    Innate immune system stimuli, such as endotoxin, seem to affect allergy risk. Previously, we described gene-environment interactions between the endotoxin receptor polymorphism C-260T of the CD14 gene and endotoxin exposure on total serum IgE level; however, the mechanism of this interaction is not known. To examine whether this gene-environment interaction affects early CD4(+)Foxp3(-) or CD4(+)Foxp3(+) lymphocyte numbers. Participating children were part of a birth cohort in the Detroit metropolitan area. Participants were genotyped for the CD14 C-260T polymorphism. Endotoxin exposure was estimated from dust measured in the home when children were 6 months old. Intracellular Foxp3 protein expression, a regulatory T-cell marker, was used to characterize CD4(+) lymphocytes in blood samples collected at the age of 12 months; total serum IgE level was also measured at this time. Because race/ethnicity may confound or modify genetic associations, all analyses were stratified by race/ethnicity. We observed a significant gene-environment interaction between CD14 C-260T genotype and endotoxin exposure on CD4(+) lymphocyte numbers, particularly CD4(+)Foxp3(-) lymphocytes. Stratified analyses suggest effect modification by race/ ethnicity on CD4(+)Foxp3(+) lymphocyte numbers, with a significant interaction in African American children but not in white children. The interaction between CD14 C-260T genotype and endotoxin exposure on total IgE levels was opposite that observed for CD4(+) lymphocyte numbers, suggesting reciprocal relationships. A gene-environment interaction between endotoxin and CD14 C-260T genotype on IgE levels may be the result of an upstream, opposing effect on CD4(+)Foxp3(+) and CD4(+)Foxp3(-) lymphocyte numbers. Race/ethnicity may affect which of these cell populations is affected by this gene-environment interaction.

  15. Production and properties of cyanobacterial endotoxins.

    Science.gov (United States)

    Keleti, G; Sykora, J L

    1982-01-01

    Lipopolysaccharides (LPS) were isolated from four species of cyanobacteria (Anabaena flos-aquae UTEX 1444. A. cylindrica, Oscillatoria tenuis, and O. brevis) frequently occurring in drinking-water supplies. The cyanobacterial LPS contained glucose, xylose, mannose, and rhamnose, but differed from the LPS derived from most gram-negative bacteria because of the variable presence of 2-keto-3-deoxyoctonate, heptose, galactose, and glucosamine. Cyanobacterial lipid A is characterized by long-chain saturated an unsaturated fatty acids and hydroxy fatty acids which show great diversity. Unlike lipid A from heterotrophic gram-negative bacteria, lipid A from cyanobacteria usually lacks phosphates. The detection of distinct O-antigen chemotypes indicates that LPS may be used for taxonomic classification. Isolated cyanobacterial LPS always induced Limulus amoebocyte lysate gelation. A. flos-aquae LPS gave a positive Schwartzman reaction. Endotoxins from A. cylindrica and O. brevis were toxic to mice when injected intraperitoneally. The cyanobacterial endotoxins showed generally lower biological activity than did LPS derived from common heterotrophic gram-negative bacteria. Nevertheless, cyanobacteria in algal blooms may be a significant source of endotoxins in water supplies. PMID:6798930

  16. Cyclic Limulus anti-lipopolysaccharide (LPS) factor-derived peptide CLP-19 antagonizes LPS function by blocking binding to LPS binding protein.

    Science.gov (United States)

    Liu, Yao; Ni, Bing; Ren, Jian-dong; Chen, Jian-hong; Tian, Zhi-qiang; Tang, Min; Li, Di; Xia, Peiyuan

    2011-01-01

    Inflammation and septic shock due to endotoxins from Gram-negative bacteria infection continue to pose significant challenges to human healthcare. It is, therefore, necessary to develop therapeutic strategies targeting endotoxins, such as lipopolysaccharide (LPS), to prevent their potentially systemic effects. Pathogenesis due to Gram-negative bacteria involves LPS binding to the host LPS-binding protein (LBP), causing detrimental downstream signaling cascades. Our previous study showed that CLP-19, a synthetic peptide derived from the Limulus anti-LPS factor (LALF), could effectively neutralize LPS toxicity; however, the detailed mechanisms underlying this anti-LPS effect remained unexplained. Thus, we carried out investigations to determine how the CLP-19 neutralizes LPS toxicity. CLP-19 was found to block LPS binding to LBP in a dose-dependent manner, as evidenced by competitive enzyme-linked immunosorbent assay (ELISA). In peripheral blood mononuclear cells, CLP-19 blocked LPS-induced phosphorylation of mitogen activated protein kinase (MAPK) signaling proteins p38, extracellular signal-regulating kinase (ERK)1/2 and c-Jun N-terminal kinase (JNK)1/2. Furthermore, CLP-19 potency in LPS antagonism in vitro and in vivo was directly associated with its ability to block the LPS-LBP interaction. Taken together, the results suggested that CLP-19's inhibitory effect on LPS-LBP binding and on the subsequent MAPK pathway signaling may be responsible for its anti-LPS mechanism. This peptide appears to represent a potential therapeutic agent for clinical treatment of sepsis.

  17. Oral contraceptives worsen endotoxin-induced liver injury in rats.

    Science.gov (United States)

    Konno, Akira; Enomoto, Nobuyuki; Takei, Yoshiyuki; Hirose, Miyoko; Ikejima, Kenichi; Sato, Nobuhiro

    2002-08-01

    Oral contraceptives are widely used; however, these drugs occasionally cause liver injury. Recently, it was reported that estriol worsens alcoholic liver injury by the mechanism involving activation of Kupffer cells as a result of gut-derived endotoxin. However, the relationship between oral contraceptives and endotoxin-induced liver injury has not been elucidated. Here we show that oral contraceptives sensitize Kupffer cells via a mechanism dependent on increased gut permeability to endotoxin. Female Wistar rats (200-250 g) were given intraperitoneally a combination of estradiol (35 ng/kg of 17 alpha-Ethynylestradiol) and progesterone (2 microg/kg of Norethindrone), each dose being similar to that contained in oral contraceptives (EP treatment). After 24 hr, a sublethal dose of lipopolysaccharide (LPS; 5 mg/kg) was injected via the tail vein. In some experiments, antibiotics (150 mg/kg/day of polymyxin B and 450 mg/kg/day of neomycin) were administered orally for 4 days before EP treatment. Gut permeability was measured in isolated segments of ileum by translocation of horseradish peroxidase. Kupffer cells were isolated and cultured in RPMI 1640 + 10% fetal bovine serum for 24 hr. After addition of LPS (100 ng/ml) to the culture medium, intracellular calcium concentration ([Ca2+](i) ) was measured with fura-2. Liver histology in rats given EP treatment intraperitoneally followed by an injection of LPS (5 mg/kg) 24 hr later revealed pronounced liver damage with massive necrosis. Whereas mean values of alanine aminotransferase (ALT) in the control, nontreated rats were 30 +/- 6 IU/liter, ALT increased to 75 +/- 21 IU/liter 24 hr after LPS injection. This increase was aggravated 6-fold (483 +/- 118 IU/liter; p< 0.05) by EP treatment. The EP treatment-induced increase in ALT was completely blocked by antibiotics (82 +/- 26 IU/liter; p< 0.05). Gut permeability was increased approximately 10-fold with EP treatment. This increase in gut permeability was not altered by

  18. Effects of Puerariae Radix Extract on Endotoxin Receptors and TNF-α Expression Induced by Gut-Derived Endotoxin in Chronic Alcoholic Liver Injury

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    Jing-Hua Peng

    2012-01-01

    Full Text Available Kudzu (Pueraria lobata is one of the earliest medicinal plants used to treat alcohol abuse in traditional Chinese medicine for more than a millennium. However, little is known about its effects on chronic alcoholic liver injury. Therefore, the present study observed the effects of puerariae radix extract (RPE on chronic alcoholic liver injury as well as Kupffer cells (KCs activation to release tumor necrosis factor alpha (TNF-α induced by gut-derived endotoxin in rats and macrophage cell line. RPE was observed to alleviate the pathological changes and lipids deposition in liver tissues as well as the serum alanine aminotransferase (ALT, aspartate aminotransferase (AST, and hepatic gamma-glutamyl transpeptidase (GGT activity. Meanwhile, RPE inhibited KCs activation and subsequent hepatic TNF-α expression and downregulated the protein expression of endotoxin receptors, lipopolysaccharide binding protein (LBP, CD14, Toll-like receptor (TLR 2, and TLR4 in chronic alcohol intake rats. Furthermore, an in vitro study showed that RPE inhibited the expression of TNF-α and endotoxin receptors, CD14 and TLR4, induced by LPS in RAW264.7 cells. In summary, this study demonstrated that RPE mitigated liver damage and lipid deposition induced by chronic alcohol intake in rats, as well as TNF-α release, protein expression of endotoxin receptors in vivo or in vitro.

  19. The injured liver induces hyperimmunoglobulinemia by failing to dispose of antigens and endotoxins in the portal system.

    Science.gov (United States)

    Liu, Wen Ting; Jing, Ying Ying; Han, Zhi Peng; Li, Xiao Ning; Liu, Yan; Lai, Fo Bao; Li, Rong; Zhao, Qiu-Dong; Wu, Meng-Chao; Wei, Li-Xin

    2015-01-01

    Hyperimmunoglobulinemia is frequently observed in patients with chronic liver diseases. However, the exact mechanism underlying the high level of antibody formation is not fully understood. In our study, we provide evidence for the functional role of the liver and the stimulation of plasma cell proliferation in hyperimmunoglobulinemia. We collected sera from patients with chronic liver diseases, and the level of serum immunoglobulins in patients was examined; this was also investigated in animal models of liver cirrhosis and hepatocellular carcinoma. An end-to-side microsurgical portacaval shunt was used to mimic liver dysfunction in rats. We used portal vein serum and inferior vena cava serum to immunize healthy rats and mice in order to confirm the function of the healthy liver in disposing of antigens and endotoxins from the gut. For the analysis of the state of plasma cell activation, plasma cells from mice were stained with PE-conjugated anti-CD138 and FITC-conjugated anti-BrdU for flow cytometry analysis. Hyperimmunoglobulinemia was observed both in patients with chronic liver diseases and in related animal models, and high plasma LPS levels were also observed. There was a significant increase in the activation and proliferation of plasma cell in mice immunized with antigens or LPS-positive serum compared with controls that were immunized with antigens and LPS-negative serum. We confirmed that the healthy liver plays an important role in disposing of antigens and endotoxins derived from the gut. Hyperimmunoglobulinemia in chronic liver diseases mainly arises due to the collateral circulation secondary to portal hypertension, gut antigens and endotoxins that bypass the liver and reach the antibody-producing cells.

  20. Bacterial endotoxin adhesion to different types of orthodontic adhesives

    Directory of Open Access Journals (Sweden)

    Priscilla Coutinho ROMUALDO

    Full Text Available Abstract Bacterial endotoxin (LPS adhesion to orthodontic brackets is a known contributing factor to inflammation of the adjacent gingival tissues. Objective The aim of this study was to assess whether LPS adheres to orthodontic adhesive systems, comparing two commercial brands. Material and Methods Forty specimens were fabricated from Transbond XT and Light Bond composite and bonding agent components (n=10/component, then contaminated by immersion in a bacterial endotoxin solution. Contaminated and non-contaminated acrylic resin samples were used as positive and negative control groups, respectively. LPS quantification was performed by the Limulus Amebocyte Lysate QCL-1000™ test. Data obtained were scored and subjected to the Chi-square test using a significance level of 5%. Results There was endotoxin adhesion to all materials (p0.05. There was no significant difference (p>0.05 among commercial brands. Affinity of endotoxin was significantly greater for the bonding agents (p=0.0025. Conclusions LPS adhered to both orthodontic adhesive systems. Regardless of the brand, the endotoxin had higher affinity for the bonding agents than for the composites. There is no previous study assessing the affinity of LPS for orthodontic adhesive systems. This study revealed that LPS adheres to orthodontic adhesive systems. Therefore, additional care is recommended to orthodontic applications of these materials.

  1. α-Solanine Isolated From Solanum Tuberosum L. cv Jayoung Abrogates LPS-Induced Inflammatory Responses Via NF-κB Inactivation in RAW 264.7 Macrophages and Endotoxin-Induced Shock Model in Mice.

    Science.gov (United States)

    Shin, Ji-Sun; Lee, Kyoung-Goo; Lee, Hwi-Ho; Lee, Hae Jun; An, Hyo-Jin; Nam, Jung-Hwan; Jang, Dae Sik; Lee, Kyung-Tae

    2016-10-01

    α-Solanine, a trisaccharide glycoalkaloid, has been reported to possess anti-cancer effects. In this study, we investigated the anti-inflammatory effects of α-solanine isolated from "Jayoung" a dark purple-fleshed potato by examining its in vitro inhibitory effects on inducible nitric-oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and pro-inflammatory cytokines in LPS-induced RAW 264.7 macrophages and its in vivo effects on LPS-induced septic shock in a mouse model. α-Solanine suppressed the expression of iNOS and COX-2 both at protein and mRNA levels and consequently inhibited nitric oxide (NO) and prostaglandin E2 (PGE2 ) production in LPS-induced RAW 264.7 macrophages. α-Solanine also reduced the production and mRNA expression of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) induced by LPS. Furthermore, molecular mechanism studies indicated that α-solanine inhibited LPS-induced activation of nuclear factor-κB (NF-κB) by reducing nuclear translocation of p65, degradation of inhibitory κBα (IκBα), and phosphorylation of IκB kinaseα/β (IKKα/β). In an in vivo experiment of LPS-induced endotoxemia, treatment with α-solanine suppressed mRNA expressions of iNOS, COX-2, IL-6, TNF-α, and IL-1β, and the activation of NF-κB in liver. Importantly, α-solanine increased the survival rate of mice in LPS-induced endotoxemia and polymicrobial sepsis models. Taken together, our data suggest that the α-solanine may be a promising therapeutic against inflammatory diseases by inhibiting the NF-κB signaling pathway. J. Cell. Biochem. 117: 2327-2339, 2016. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  2. Treatment of Lipoxin A(4) and its analogue on low-dose endotoxin induced preeclampsia in rat and possible mechanisms.

    Science.gov (United States)

    Lin, Feng; Zeng, Pan; Xu, Zhangye; Ye, Duyun; Yu, Xiaofang; Wang, Ni; Tang, Jie; Zhou, Ye; Huang, Yinping

    2012-12-01

    Preeclampsia (PE) is known to represent an exaggerated maternal inflammatory response to pregnancy. Lipoxin A(4) (LXA(4)), considered as an endogenous stop signal in inflammation, has been extensively studied pre clinically for its inflammatory pro-resolving effects. Thus, in the current study, we tested the effect of BML-111 (synthetic analogue of LXA(4)) on experimental PE rats induced by low-dose endotoxin (LPS) and of LXA(4) on human extravillous trophoblast cell line (TEV-1). In vivo experiment results showed that systolic blood pressure, 24 h-urinary albumin excretion, serum TNF-α and IL-8 levels and morphologic damage of placenta and kidney caused by LPS were all effectively alleviated by BML-111. LXA(4) also inhibited LPS-triggered apoptosis, activation of NF-κB, TNF-α and IL-8 mRNA and protein expression in TEV-1 cells. At the same time, BML-111 protected the cells from LPS-reduced proliferation. The current study demonstrated for the first time that LXA(4) could alleviate the symptoms of PE in endotoxin exposed rats. Copyright © 2012 Elsevier Inc. All rights reserved.

  3. LPS from bovine serum albumin drives TNF-α release during ex-vivo placenta perfusion experiments, contaminates the perfusion system but can be effectively removed by oxidative cleaning.

    Science.gov (United States)

    Vasanthan, T; Rochow, N; Mian, F; Codini, T; DeFrance, B; Fusch, G; Samiee-Zafarghandy, S; Fusch, C

    2014-12-01

    The dual ex-vivo perfusion of human placental tissue is useful to study inflammatory pathways. We found significant TNF-α release in negative controls similar in concentration to lipopolysaccharide (LPS) stimulated placentas. The aim of the current study was to (i) identify sources driving TNF-α release and (ii) develop an approach to control for it. (i) To determine sources leading to TNF-α release, solutions frequently circulated through the perfusion system and perfusion media with different bovine serum albumin (BSA) quality were exposed to mouse macrophage cell lines (RAW264.7) and subsequently measured for TNF-α expression. (ii) To assess memory effects and validate cleaning procedures, sham perfusion experiments were conducted either in the presence or absence of exogenous LPS, in new tubing that was contaminated, cleaned and analyzed for the effectiveness of LPS removal. Oxidative and acid-base cleaning were tested for their effectiveness to reduce LPS contamination. TNF-α release, observed in negative control experiments, was attributed to the use of LPS-contaminated BSA as well as inadequate cleaning of the perfusion system. Once introduced in the perfusion system, LPS accumulated and created a memory effect. Oxidative but not acid-base depyrogenation effectively reduced LPS levels to concentrations that were in accordance with FDA guidelines (contamination of the placenta perfusion model could have confounding effects on experimental outcomes leading to misinterpretation of data. To circumvent LPS contamination LPS-free BSA and oxidative depyrogenation cleaning techniques should be implemented in future placental perfusion studies. Copyright © 2014 Elsevier Ltd. All rights reserved.

  4. Pseudoalteromonas strains are potent immunomodulators owing to low-stimulatory LPS

    DEFF Research Database (Denmark)

    Maaetoft-Udsen, Kristina; Vynne, Nikolaj Grønnegaard; Heegaard, Peter M. H.

    2013-01-01

    confirmed the involvement of LPS and TLR4 and established Pseudoalteromonas LPS as TLR4 antagonists. The isolated LPS was active in the endotoxin limulus amoebocyte lysate assay and capable of inducing increased endocytosis in DCs. This study highlights that antagonistic LPS from Pseudoalteromonas strains...... investigated with respect to their immunomodulatory properties in DCs. Both inhibited IL-12 and increased IL-10 production induced by E. coli LPS. LPS isolated from the two Pseudoalteromonas strains had characteristic lipid A bands in SDS-PAGE. Stimulation of HEK293 TLR4/MD2 cells with the isolated LPS...

  5. Endotoxin administration to humans inhibits hepatic cytochrome P450-mediated drug metabolism.

    Science.gov (United States)

    Shedlofsky, S I; Israel, B C; McClain, C J; Hill, D B; Blouin, R A

    1994-12-01

    In experimental animals, injection of gram-negative endotoxin (LPS) decreases hepatic cytochrome P450-mediated drug metabolism. To evaluate this phenomenon in a human model of gram-negative sepsis, LPS was administered on two consecutive days to healthy male volunteers during which time a cocktail of antipyrine (AP-250 mg), hexobarbital (HB-500 mg), and theophylline (TH-150 mg) was ingested and the apparent oral clearance of each drug determined. Each subject had a control drug clearance study with saline injections. In the first experiment, six subjects received the drug cocktail 0.5 h after the first dose of LPS. In the second experiment, another six subjects received the drug cocktail 0.5 h after the second dose of LPS. In both experiments, LPS caused the expected physiologic responses of inflammation including fever with increases in serum concentrations of TNF alpha, IL-1 beta, IL-6, and acute phase reactants. In the first experiment, only minor decreases in clearances of the probe drugs were observed (7-12%). However in the second experiment, marked decreases in the clearances of AP (35, 95% CI 18-48%), HB (27, 95% CI 14-34%), and TH (22, 95% CI 12-32%) were seen. The decreases in AP clearance correlated with initial peak values of TNF alpha (r = 0.82) and IL-6 (r = 0.86). These data show that in humans the inflammatory response to even a very low dose of LPS significantly decreases hepatic cytochrome P450-mediated drug metabolism and this effect evolves over a 24-h period. It is likely that septic patients with much higher exposures to LPS have more profound inhibition of drug metabolism.

  6. Vasorelaxing Action of the Kynurenine Metabolite, Xanthurenic Acid: The Missing Link in Endotoxin-Induced Hypotension?

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    Carmine Vecchione

    2017-05-01

    Full Text Available The kynurenine pathway of tryptophan metabolism is activated by pro-inflammatory cytokines. L-kynurenine, an upstream metabolite of the pathway, acts as a putative endothelium-derived relaxing factor, and has been hypothesized to play a causative role in the pathophysiology of inflammation-induced hypotension. Here, we show that xanthurenic acid (XA, the transamination product of 3-hydroxykynurenine, is more efficacious than L-kynurenine in causing relaxation of a resistance artery, but fails to relax pre-contracted aortic rings. In the mesenteric artery, XA enhanced activating phosphorylation of endothelial nitric oxide synthase (NOS, and the relaxing action of XA was abrogated by pharmacological inhibition of NOS and endothelial-derived hyperpolarizing factor. Systemic injection of XA reduced blood pressure in mice, and serum levels of XA increased by several fold in response to a pulse with the endotoxin, lipopolysaccharide (LPS. LPS-induced hypotension in mice was prevented by pre-treatment with the kynurenine monooxygenase (KMO inhibitor, Ro-618048, which lowered serum levels of XA but enhanced serum levels of L-kynurenine. UPF 648, another KMO inhibitor, could also abrogate LPS-induced hypotension. Our data identify XA as a novel vasoactive compound and suggest that formation of XA is a key event in the pathophysiology of inflammation-induced hypotension.

  7. Effect of the Toll-Like Receptor 4 Antagonist Eritoran on Retinochoroidal Inflammatory Damage in a Rat Model of Endotoxin-Induced Inflammation

    Directory of Open Access Journals (Sweden)

    Feyzahan Ekici

    2014-01-01

    Full Text Available Purpose. We investigated the effect of eritoran, a Toll-like receptor 4 antagonist, on retinochoroidal inflammatory damage in an endotoxin-induced inflammatory rat model. Methods. Endotoxin-induced inflammatory model was obtained by intraperitoneal injection of 1.5 mg/kg lipopolysaccharide (LPS. Group 1 had control rats; in groups 2-3 LPS and 0.5 mg/kg sterile saline were injected; and in groups 4-5 LPS and 0.5 mg/kg eritoran were injected. Blood samples were taken and eyes were enucleated after 12 hours (h (groups 2 and 4 or 24 hours (Groups 3 and 5. Tumor necrosis factor-α (TNF-α and malondialdehyde (MDA levels in the serum and retinochoroidal tissue and nuclear factor kappa-B (NFκB levels in retinochoroidal tissue were determined. Histopathological examination was performed and retinochoroidal changes were scored. Results. Eritoran treatment resulted in lower levels of TNF-α, MDA, and NFκB after 12 h which became significant after 24 h. Serum TNF-α and retinochoroidal tissue NFκB levels were similar to control animals at the 24th h of the study. Eritoran significantly reversed histopathological damage after 24 h. Conclusions. Eritoran treatment resulted in less inflammatory damage in terms of serum and retinochoroidal tissue parameters.

  8. Neutralization of bacterial endotoxins by frog antimicrobial peptides.

    Science.gov (United States)

    Schadich, Ermin; Mason, Drusilla; Cole, Anthony L

    2013-02-01

    The ability of skin antimicrobial peptides of the southern bell frog, Litoria raniformis, to neutralize in vitro the endotoxin, proinflammatory lipopolysaccharide (LPS) complex, from two different gram-negative bacterial pathogens, human pathogen Escherichia coli (0111:B4) and frog pathogen Klebsiella pneumoniae, was investigated. The LPS neutralization activity of the natural mixture of skin antimicrobial peptides was measured using chromogenic Limulus amebocyte lysate assays. These skin antimicrobial peptides neutralized the LPSs from both pathogens at physiologically relevant concentrations (IC(50)  endotoxin agents. © 2012 The Societies and Wiley Publishing Asia Pty Ltd.

  9. Biophysical investigations into the interactions of endotoxins with bile acids.

    Science.gov (United States)

    Fukuoka, Satoshi; Richter, Walter; Howe, Jörg; Andrä, Jörg; Rössle, Manfred; Alexander, Christian; Gutsmann, Thomas; Brandenburg, Klaus

    2012-04-01

    The interaction of selected endotoxin preparations (lipid A from Erwinia carotovora and LPS Re and Ra from Salmonella enterica sv. Minnesota strains R595 and R60, respectively) with selected bile acids was investigated biophysically. Endotoxin aggregates were analyzed for their gel-to-liquid crystalline phase behavior, the type of their aggregates, the conformation of particular functional groups, and their Zeta potential in the absence and presence of the bile acids by applying Fourier-transform infrared spectroscopy, differential scanning calorimetry, measurements of the electrophoretic mobility, and synchrotron radiation X-ray scattering. In addition, the ability of the endotoxins to induce cytokines in human mononuclear cells was tested in the absence and presence of varying concentrations of bile acids. The data show that the endotoxin:bile acid interaction is not governed by Coulomb forces, rather a hydrophobic interaction takes place. This leads to an enhanced formation of the inherent cubic aggregate structures of the endotoxins, concomitant with a slight disaggregation, as evidenced by freeze-fracture electron microscopy. Parallel to this, the addition of bile acids increased the bioactivity of lipid A and, to a lower degree, also that of the tested rough mutant LPS at lower concentrations of the endotoxin preparation, a finding similar as reported for the interaction of other agents such as hemoglobin. These data imply that there are general mechanisms that govern the expression of biological activities of endotoxins.

  10. Endotoxin tolerance alleviates experimental acute liver failure via inhibition of high mobility group box 1.

    Science.gov (United States)

    Yang, Nai-Bin; Ni, Shun-Lan; Li, Shan-Shan; Zhang, Sai-Nan; Hu, Dan-Ping; Lu, Ming-Qin

    2015-01-01

    High mobility group box 1 (HMGB1) has been widely reported to mediate damage caused by inflammatory responses. The aim of our study is to investigate the role of HMGB1 in endotoxin tolerance (ET) alleviating inflammation of acute liver failure (ALF) rats and its possible signaling mechanism. To mimic ET, male Sprague-Dawley rats were pretreated with low dose of lipopolysaccharide (LPS) (0.1 mg/kg once a day intraperitoneally for consecutive five days) before subsequent ALF induction. ALF was induced by intraperitoneal administration of D-GalN/LPS. ET induced by LPS pretreatment significantly improved the survival rate of ALF rats. Moreover, after ALF induction, ET+ALF rats exhibited lower serum enzyme (ALT, AST and TBiL) levels, lower production of inflammatory cytokines (IL-6, TNF-a and HMGB1) and more minor liver histopathological damage than ALF rats. ET+ALF rats showed enhanced expression levels of HMGB1, decreased levels of STAT1 and p-STAT1, augmented expression of SOCS1 in liver tissues than ALF rats. These results indicated that ET induced by low-dose LPS pretreatment may alleviate inflammation and liver injury in experimental acute liver failure rats mainly through inhibition of hepatic HMGB1 translocation and release.

  11. Effects of duration of moderate increases in grain feeding on endotoxins in the digestive tract and acute phase proteins in peripheral blood of yearling calves.

    Science.gov (United States)

    Plaizier, J C; Li, S; Le Sciellour, M; Schurmann, B L; Górka, P; Penner, G B

    2014-11-01

    Effects of duration of grain feeding on the concentration of endotoxic lipopolysaccharide (LPS) in digesta throughout the digestive tract and on acute phase proteins and LPS in peripheral blood were determined in Holstein yearling calves. Twenty-five Holstein yearling steer calves received either a forage-based diet containing 92% hay and 8% of a mineral and vitamin pellet on a dry matter basis (CON) or a moderate-grain diet, obtained by replacing 41.5% of the hay in the forage-based diet with barley grain, for 3 (MG3), 7 (MG7), 14 (MG14), or 21 d (MG21) before slaughter. Immediately before slaughter, blood samples were collected from the jugular vein. Immediately after slaughter, digesta samples were collected from the rumen, jejunum, ileum, cecum, colon, and rectum. Rumen liquid digesta, digesta from the intestines, and peripheral blood plasma were analyzed for LPS. Peripheral blood plasma and serum were analyzed for the acute phase proteins amyloid A, haptoglobin, and LPS-binding protein. Feeding the grain diet increased the LPS concentration in rumen fluid linearly from 15,488 endotoxin units (EU)/mL for CON to 70,146 EU/mL for MG7. Concentrations of LPS in rumen fluid in MG14 and MG21 were 61,944 and 56,234 EU/mL, respectively, and did not differ. The LPS concentrations in jejunal digesta were much lower than that in digesta elsewhere in the digestive tract, which suggests that ruminal LPS is broken down in the abomasum or proximal jejunum. The concentration of digesta LPS in the ileum was higher than that of digesta elsewhere in the intestines and similar to that in rumen fluid. The duration of grain feeding increased the LPS concentration in digesta in the ileum and cecum and tended to increase that in the colon cubically. Concentrations of LPS in this part of the digestive tract were highest in the MG3 and MG21 groups. The highest concentrations of LPS in digesta in the cecum, colon, and rectum were 3.7, 3.8, and 5.6 times higher than that in CON

  12. Critical aggregation concentrations of gram-negative bacterial lipopolysaccharides (LPS).

    Science.gov (United States)

    Aurell, C A; Wistrom, A O

    1998-12-09

    Bacterial endotoxins or lipopolysaccharides (LPS), cell wall components of gram negative bacteria, are involved in septic shock. While the carbohydrate structure of LPS have been studied in the past, little is known about the macromolecular structure and formation of LPS fragments in blood. It is believed that amphiphilic molecules such as LPS occur as monomers and aggregate into macromolecular structures above a critical micelle or critical aggregate concentration, CAC. The CAC of Lipid A, a LPS precursor, and several LPS serotypes of varying molecular weight and different polysaccharide chain lengths were established by static light scattering and by steady-state fluorescence spectroscopy by incorporation of the fluorescent probe, NPN, N-phenyl-1-naphthylamine. The CAC for short polysaccharide chain mutant LPS S.minnesota Re 595 (MW 2,500) and Lipid A from S.minnesota Re 595 (MW 2000) were 4 microM and 5 microM respectively. The CAC of LPS from heterogeneous long O-antigen polysaccharide chain bacterial serotypes: S.minnesota wildtype were 11 micrograms LPS/ml, S.typhimurium 14 micrograms LPS/ml and E.coli 0111:B4 22 micrograms LPS/ml, respectively. The results obtained suggests that critical aggregate concentration and solubility of LPS is a function of polysaccharide chain length.

  13. Portal triad occlusion induces endotoxin tolerance: role of portal congestion.

    Science.gov (United States)

    Unno, Naoki; Uchiyama, Takashi; Yamamoto, Naoto; Inuzuka, Kazunori; Sagara, Daisuke; Konno, Hiroyuki

    2006-10-01

    Portal triad occlusion (PTO) causes portal congestion and damages the intestinal mucosa, which is associated with portal endotoxemia. However, administration of a sublethal dose of endotoxin results in resistance to its toxic activities. We tested the hypothesis that portal congestion due to PTO induces endotoxin tolerance. Rats were subjected to PTO for 15 min. In Group 1, male rats underwent laparotomy and, 48 h after the surgery, a lethal dose of Escherichia coli lipopolysaccharide was administered. In Group 2, rats were subjected to PTO for 15 min. Then a lethal dose of LPS was administered 48 h after surgery. Group 3 was treated the same as Group 2, except that PTO was performed with portosystemic shunt. Group 4 was also treated same as Group 2, except that rats received polymixin B and neomycin by gavage to eliminate intestinal luminal bacteria before PTO. Survival was examined after the administration of a lethal dose of LPS. Changes in plasma levels of cytokine are also measured after the administration of LPS. The portal endotoxin level in each group after PTO was measured. On survival test, only rats in Group 2 and Group 4 showed significantly higher survival rates. The portal endotoxin level was significantly elevated only in Group 2. The elevation of plasma cytokine levels (IL-6, TNF-alpha) and NO production (NO(2)(-)/NO(3)(-)) in Groups 2 and 4 were inhibited compare to those in Groups 1 and 3. PTO induced LPS tolerance possibly due to portal congestion and subsequent visceral congestion.

  14. Biophysical Analysis of Lipopolysaccharide Formulations for an Understanding of the Low Endotoxin Recovery (LER Phenomenon

    Directory of Open Access Journals (Sweden)

    Wilmar Correa

    2017-12-01

    Full Text Available Lipopolysaccharides (LPS, endotoxin are complex and indispensable components of the outer membrane of most Gram-negative bacteria. They represent stimuli for many biological effects with pathophysiological character. Recombinant therapeutic proteins that are manufactured using biotechnological processes are prone to LPS contaminations due to their ubiquitous occurrence. The maximum endotoxin load of recombinant therapeutic proteins must be below the pyrogenic threshold. Certain matrices that are commonly used for recombinant therapeutic proteins show a phenomenon called “Low Endotoxin Recovery (LER”. LER is defined as the loss of detectable endotoxin activity over time using compendial Limulus amebocyte lysate (LAL assays when undiluted products are spiked with known amount of endotoxin standards. Because LER poses potential risks that endotoxin contaminations in products may be underestimated or undetected by the LAL assay, the United States (U.S. Food and Drug Administration’s (FDA’s Center for Drug Evaluation and Research (CDER has recently started requesting that companies conduct endotoxin spike/hold recovery studies to determine whether a given biological product causes LER. Here, we have performed an analysis of different LPS preparations with relevant detergents studying their acyl chain phase transition, their aggregate structures, their size distributions, and binding affinity with a particular anti-endotoxin peptide, and correlating it with the respective data in the macrophage activation test. In this way, we have worked out biophysical parameters that are important for an understanding of LER.

  15. Contamination of nanoparticles by endotoxin: evaluation of different test methods

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    Smulders Stijn

    2012-11-01

    Full Text Available Abstract Background Nanomaterials can be contaminated with endotoxin (lipopolysaccharides, LPS during production or handling. In this study, we searched for a convenient in vitro method to evaluate endotoxin contamination in nanoparticle samples. We assessed the reliability of the commonly used limulus amebocyte lysate (LAL assay and an alternative method based on toll-like receptor (TLR 4 reporter cells when applied with particles (TiO2, Ag, CaCO3 and SiO2, or after extraction of the endotoxin as described in the ISO norm 29701. Results Our results indicate that the gel clot LAL assay is easily disturbed in the presence of nanoparticles; and that the endotoxin extraction protocol is not suitable at high particle concentrations. The chromogenic-based LAL endotoxin detection systems (chromogenic LAL assay and Endosafe-PTS, and the TLR4 reporter cells were not significantly perturbed. Conclusion We demonstrated that nanoparticles can interfere with endotoxin detection systems indicating that a convenient test method must be chosen before assessing endotoxin contamination in nanoparticle samples.

  16. Maternal endotoxin-induced fetal growth restriction in rats: Fetal responses in toll-like receptor

    Directory of Open Access Journals (Sweden)

    Banun Kusumawardani

    2012-09-01

    Full Text Available Background: Porphyromonas gingivalis as a major etiology of periodontal disease can produce virulence factor, lipopolysaccharide/LPS, which is expected to play a role in the intrauterine fetal growth. Trophoblast at the maternal-fetal interface actively participates in response to infection through the expression of a family of natural immune receptors, toll-like receptor (TLR. Purpose: the aims of study were to identify endotoxin concentration in maternal blood serum of Porphyromonas gingivalis-infected pregnant rats, to characterize the TLR-4 expression in trophoblast cells, and to determine its effect on fetal growth. Methods: Female rats were infected with live-Porphyromonas gingivalis at concentration of 2 x 109 cells/ml into subgingival sulcus area of the maxillary first molar before and/or during pregnancy. They were sacrified on 14th and 20th gestational day. Fetuses were evaluated for weight and length. Endotoxin was detected by limulus amebocyte lysate assay in the maternal blood serum. The TLR-4 expression in trophoblast cells was detected by immunohistochemistry. The effects of a selective inhibitor of c-Fos/activator protein-1 on endotoxin-induced acute kidney injury in mice

    Directory of Open Access Journals (Sweden)

    Miyazaki Hiroyuki

    2012-11-01

    Full Text Available Abstract Background Sepsis has been identified as the most common cause of acute kidney injury (AKI in intensive care units. Lipopolysaccharide (LPS induces the production of several proinflammatory cytokines including tumor necrosis factor (TNF-alpha, a major pathogenetic factor in septic AKI. c-Fos/activator protein (AP-1 controls the expression of these cytokines by binding directly to AP-1 motifs in the cytokine promoter regions. T-5224 is a new drug developed by computer-aided drug design that selectively inhibits c-Fos/AP-1 binding to DNA. In this study, we tested whether T-5224 has a potential inhibitory effect against LPS-induced AKI, by suppressing the TNF-alpha inflammatory response and other downstream effectors. Methods To test this hypothesis, male C57BL/6 mice at 7 weeks old were divided into three groups (control, LPS and T-5224 groups. Mice in the control group received saline intraperitoneally and polyvinylpyrrolidone solution orally. Mice in the LPS group were injected intraperitoneally with a 6 mg/kg dose of LPS and were given polyvinylpyrrolidone solution immediately after LPS injection. In the T-5224 group, mice were administered T-5224 orally at a dose of 300 mg/kg immediately after LPS injection. Serum concentrations of TNF-alpha, interleukin (IL-1beta, IL-6 and IL-10 were measured by ELISA. Moreover, the expression of intercellular adhesion molecule (ICAM-1 mRNA in kidney was examined by quantitative real-time RT-PCR. Finally, we evaluated renal histological changes. Results LPS injection induced high serum levels of TNF-alpha, IL-1beta and IL-6. However, the administration of T-5224 inhibited the LPS-induced increase in these cytokine levels. The serum levels of IL-10 in the LPS group and T-5224 group were markedly elevated compared with the control group. T-5224 also inhibited LPS-induced ICAM-1 mRNA expression. Furthermore histological studies supported an anti-inflammatory role of T-5224. Conclusions In endotoxin

  17. Regulation of Lipolysis and Adipose Tissue Signaling during Acute Endotoxin-Induced Inflammation: A Human Randomized Crossover Trial.

    Science.gov (United States)

    Rittig, Nikolaj; Bach, Ermina; Thomsen, Henrik Holm; Pedersen, Steen Bønlykke; Nielsen, Thomas Sava; Jørgensen, Jens O; Jessen, Niels; Møller, Niels

    2016-01-01

    Lipolysis is accelerated during the acute phase of inflammation, a process being regulated by pro-inflammatory cytokines (e.g. TNF-α), stress-hormones, and insulin. The intracellular mechanisms remain elusive and we therefore measured pro- and anti-lipolytic signaling pathways in adipocytes after in vivo endotoxin exposure. Eight healthy, lean, male subjects were investigated using a randomized cross over trial with two interventions: i) bolus injection of saline (Placebo) and ii) bolus injection of lipopolysaccharide endotoxin (LPS). A 3H-palmitate tracer was used to measure palmitate rate of appearance (Rapalmitate) and indirect calorimetry was performed to measure energy expenditures and lipid oxidation rates. A subcutaneous abdominal fat biopsy was obtained during both interventions and subjected to western blotting and qPCR quantifications. LPS caused a mean increase in serum free fatty acids (FFA) concentrations of 90% (CI-95%: 37-142, p = 0.005), a median increase in Rapalmitate of 117% (CI-95%: 77-166, ptrend towards elevated pHSL at ser552 (p = 0.09) and cAMP-dependent protein kinase A (PKA) phosphorylation of perilipin 1 (PLIN1) (p = 0.09). Phosphatase and tensin homolog (PTEN) also tended to increase (p = 0.08) while phosphorylation of Akt at Thr308 tended to decrease (p = 0.09) during LPS compared with Placebo. There was no difference between protein or mRNA expression of ATGL, G0S2, and CGI-58. LPS stimulated lipolysis in adipose tissue and is associated with increased pHSL and signs of increased PLIN1 phosphorylation combined with a trend toward decreased insulin signaling. The combination of these mechanisms appear to be the driving forces behind the increased lipolysis observed in the early stages of acute inflammation and sepsis. ClinicalTrials.gov NCT01705782.

  18. Defective LPS Signaling in C3H/HeJ and C57BL/10ScCr Mice: Mutations in Tlr4 Gene

    National Research Council Canada - National Science Library

    Alexander Poltorak; Xiaolong He; Irina Smirnova; Mu-Ya Liu; Christophe Van Huffel; Xin Du; Dale Birdwell; Erica Alejos; Maria Silva; Chris Galanos; Marina Freudenberg; Paola Ricciardi-Castagnoli; Betsy Layton; Bruce Beutler

    1998-01-01

    Mutations of the gene Lps selectively impede lipopolysaccharide (LPS) signal transduction in C3H/HeJ and C57BL/10ScCr mice, rendering them resistant to endotoxin yet highly susceptible to Gram-negative infection. The codominant Lps$^d...

  19. Variation in Microbiome LPS Immunogenicity Contributes to Autoimmunity in Humans

    Science.gov (United States)

    Vatanen, Tommi; Kostic, Aleksandar D.; d’Hennezel, Eva; Siljander, Heli; Franzosa, Eric A.; Yassour, Moran; Kolde, Raivo; Vlamakis, Hera; Arthur, Timothy D.; Hämäläinen, Anu-Maaria; Peet, Aleksandr; Tillmann, Vallo; Uibo, Raivo; Mokurov, Sergei; Dorshakova, Natalya; Ilonen, Jorma; Virtanen, Suvi M.; Szabo, Susanne J.; Porter, Jeffrey A.; Lähdesmäki, Harri; Huttenhower, Curtis; Gevers, Dirk; Cullen, Thomas W.; Knip, Mikael; Xavier, Ramnik J.

    2016-01-01

    Summary According to the hygiene hypothesis, the increasing incidence of autoimmune diseases in western countries may be explained by changes in early microbial exposure, leading to altered immune maturation. We followed gut microbiome development from birth until age three in 222 infants in Northern Europe, where early-onset autoimmune diseases are common in Finland and Estonia but less prevalent in Russia. We found that Bacteroides species are lowly abundant in Russians but dominate in Finnish and Estonian infants. Therefore their Lipopolysaccharide (LPS) exposures arose primarily from Bacteroides rather than from Escherichia coli which is a potent innate immune activator. We show that Bacteroides LPS is structurally distinct from E. coli LPS and inhibits innate immune signaling and endotoxin tolerance; furthermore, unlike LPS from E. coli, B. dorei LPS does not decrease incidence of autoimmune diabetes in non-obese diabetic mice. Early colonization by immunologically silencing microbiota may thus preclude aspects of immune education. PMID:27133167

  1. The effects of early allergen/endotoxin exposure on subsequent allergic airway inflammation to allergen in mouse model of asthma

    Directory of Open Access Journals (Sweden)

    Yeong-Ho Rha

    2010-04-01

    Full Text Available Purpose : Recently many studies show early exposure during childhood growth to endotoxin (lipopolysaccharides, LPS and/or early exposure to allergens exhibit important role in development of allergy including bronchial asthma. The aim of this study was to evaluate the role of endotoxin and allergen exposure in early life via the airways in the pathogenesis of allergic airways inflammation and airway hyperresposiveness (AHR in mouse model of asthma. Methods : Less than one week-old Balb/c mice was used. Groups of mice were received either a single intranasal instillation of sterile physiologic saline, 1% ovalbumin (OVA, LPS or 1.0 μg LPS in 1% OVA. On 35th day, these animals were sensitized with 1% OVA for 10 consecutive days via the airways. Animals were challenged with ovalbumin for 3 days on 55th days, and airway inflammation, hyperresponsiveness, and cytokine expression were assessed. Measurements of airway function were obtained in unrestrained animals, using whole-body plethysmography. Airway responsiveness was expressed in terms of % enhanced pause (Penh increase from baseline to aerosolized methacholine. Lung eosinophilia, serum OVA-IgE and bronchoalveolar lavage (BAL fluid cytokine levels were also assessed. ANOVA was used to determine the levels of difference between all groups. Comparisons for all pairs were performed by Tukey-Kramer honest significant difference test; P values for significance were set to 0.05. Results : Sensitized and challenged mice with OVA showed significant airway eosinophilia and heightened responsiveness to methacholine. Early life exposure of OVA and/or LPS via the airway prevented both development of AHR as well as bronchoalveolar lavage fluid eosinophilia. Exposure with OVA or LPS also resulted in suppression of interleukin (IL-4, 5 production in BAL fluid and OVA specific IgE in blood. Conclusion : These results indicate that antigen and/or LPS exposure in the early life results in inhibition of allergic

  2. Allergen specific responses in cord and adult blood are differentially modulated in the presence of endotoxins.

    Science.gov (United States)

    Eiwegger, T; Mayer, E; Brix, S; Schabussova, I; Dehlink, E; Bohle, B; Barkholt, V; Szépfalusi, Z

    2008-10-01

    Endotoxins are common contaminants in allergen preparations and affect antigen-specific cellular responses. Distinct effects of endotoxin on cells in human umbilical cord and adult blood are poorly defined. To examine the effect of endotoxins in allergen preparations on cellular responses in human cord and peripheral blood (PB). The endotoxin content in beta lactoglobulin (BLG), the peanut allergen Ara h 1 and the major birch pollen allergen Bet v 1 was assessed. Proliferation and cytokine response of mononuclear cells towards contaminated and lipopolysaccharide (LPS)-free allergens were evaluated at different time-points. Fractions of contaminated BLG were generated and assayed on their immuno-stimulatory capacity. The involvement of toll-like receptor (TLR) 2 and 4 was investigated by blocking antibodies and TLR-transfected human embryonic kidney cells. The proliferative response of cord blood (CB)-derived mononuclear cells towards allergen-preparations at day 3 was related to the level of LPS contamination. At day 7, proliferation was also detected in the absence of endotoxin. Cytokine production in CB was strongly affected by the content of endotoxin, TLR-4 dependent and not related to the allergen content. Allergen- and endotoxin-induced proliferative responses were generally significantly higher in CB than in adult blood. Endotoxins in allergen preparations confound allergen-specific cellular responses. The impact of these contaminations varies with the blood source (CB vs. PB), the type of allergen and is time- and dose-dependent.

  3. Effects of hepatocyte CD14 upregulation during cholestasis on endotoxin sensitivity.

    Directory of Open Access Journals (Sweden)

    Ming-Huei Chou

    Full Text Available Cholestasis is frequently related to endotoxemia and inflammatory response. Our previous investigation revealed a significant increase in plasma endotoxin and CD14 levels during biliary atresia. We therefore propose that lipopolysacharides (LPS may stimulate CD14 production in liver cells and promote the removal of endotoxins. The aims of this study are to test the hypothesis that CD14 is upregulated by LPS and investigate the pathophysiological role of CD14 production during cholestasis. Using Western blotting, qRT-PCR, and promoter activity assay, we demonstrated that LPS was associated with a significant increase in CD14 and MD2 protein and mRNA expression and CD14 promoter activity in C9 rat hepatocytes but not in the HSC-T6 hepatic stellate cell line in vitro. To correlate CD14 expression and endotoxin sensitivity, in vivo biliary LPS administration was performed on rats two weeks after they were subjected to bile duct ligation (BDL or a sham operation. CD14 expression and endotoxin levels were found to significantly increase after LPS administration in BDL rats. These returned to basal levels after 24 h. In contrast, although endotoxin levels were increased in sham-operated rats given LPS, no increase in CD14 expression was observed. However, mortality within 24 h was more frequent in the BDL animals than in the sham-operated group. In conclusion, cholestasis and LPS stimulation were here found to upregulate hepatic CD14 expression, which may have led to increased endotoxin sensitivity and host proinflammatory reactions, causing organ failure and death in BDL rats.

  4. The redistribution of granulocytes following E. coli endotoxin induced sepsis

    DEFF Research Database (Denmark)

    Toft, P; Lillevang, S T; Tønnesen, Else Kirstine

    1994-01-01

    Infusion of endotoxin elicits granulocytopenia followed by increased numbers of granulocytes in peripheral blood. The purpose of this study was to investigate the redistribution and sequestration of granulocytes in the tissues following E. coli endotoxin induced sepsis. From 16 rabbits granulocytes...... were isolated, labelled with Indium and reinjected intravenously. Eight rabbits received an infusion of E. coli endotoxin 2 micrograms kg-1 while eight received isotonic saline. The redistribution of granulocytes was imaged with a gamma camera and calculated with a connected computer before and 2 and 6...... hours after infusion of endotoxin or saline. Serum cortisol and interleukin-1 beta were measured. In another seven rabbits, respiratory burst activity and degranulation of granulocytes were measured prior to and from 5 min to 6 hours after infusion of E. coli endotoxin 2 micrograms kg-1 BW. Following...

  5. Effects of Aging on Endotoxin Tolerance Induced by Lipopolysaccharides Derived from Porphyromonas gingivalis and Escherichia coli

    OpenAIRE

    Ying Sun; Hui Li; Mi-Fang Yang; Wei Shu; Meng-Jun Sun; Yan Xu

    2012-01-01

    BACKGROUND: Periodontitis is a bacterially induced chronic inflammatory disease. Exposure of the host to periodontal pathogens and their virulence factors induces a state of hyporesponsiveness to subsequent stimulations, termed endotoxin tolerance. Aging has a profound effect on immune response to bacteria challenge. The aim of this study was to explore the effects of aging on endotoxin tolerance induced by Porphyromonas gingivalis (P. gingivalis) lipopolysaccharide (LPS) and Escherichia coli...

  6. D-TRP(8-γMSH Prevents the Effects of Endotoxin in Rat Skeletal Muscle Cells through TNFα/NF-KB Signalling Pathway.

    Directory of Open Access Journals (Sweden)

    Ana Belén Gómez-SanMiguel

    Full Text Available Sepsis induces anorexia and muscle wasting secondary to an increase in muscle proteolysis. Melanocyte stimulating hormones (MSH is a family of peptides that have potent anti-inflammatory effects. Melanocortin receptor-3 (MC3-R has been reported as the predominant anti-inflammatory receptor for melanocortins. The aim of this work was to analyse whether activation of MC3-R, by administration of its agonist D-Trp(8-γMSH, is able to modify the response of skeletal muscle to inflammation induced by lipopolysaccharide endotoxin (LPS or TNFα. Adult male rats were injected with 250 μg/kg LPS and/or 500 μg/kg D-Trp(8-γMSH 17:00 h and at 8:00 h the following day, and euthanized 4 hours afterwards. D-Trp(8-γMSH decreased LPS-induced anorexia and prevented the stimulatory effect of LPS on hypothalamic IL-1β, COX-2 and CRH as well as on serum ACTH and corticosterone. Serum IGF-I and its expression in liver and gastrocnemius were decreased in rats injected with LPS, but not in those that also received D-Trp(8-γMSH. However, D-Trp(8-γMSH was unable to modify the effect of LPS on IGFBP-3. In the gastrocnemius D-Trp(8-γMSH blocked LPS-induced decrease in pAkt, pmTOR, MHC I and MCH II, as well as the increase in pNF-κB(p65, FoxO1, FoxO3, LC3b, Bnip-3, Gabarap1, atrogin-1, MuRF1 and in LC3a/b lipidation. In L6 myotube cultures, D-Trp(8-γMSH was able to prevent TNFα-induced increase of NF-κB(p65 phosphorylation and decrease of Akt phosphorylation as well as of IGF-I and MHC I expression. These data suggest that MC3-R activation prevents the effect of endotoxin on skeletal wasting by modifying inflammation, corticosterone and IGF-I responses and also by directly acting on muscle cells through the TNFα/NF-κB(p65 pathway.

  7. Endotoxins in commercial vaccines.

    Science.gov (United States)

    Geier, M R; Stanbro, H; Merril, C R

    1978-01-01

    Twenty samples of commercial vaccines intended for administration to humans were assayed for the presence of bacterial endotoxins by using the Limulus amebocyte lysate test. Sixteen of the vaccines contained more than 0.1 ng of endotoxin per ml (which corresponds to 103 bacterial cell wall equivalents per ml in the undiluted vaccines). These results suggest that at some stage of preparation, the vaccines have contained varying amounts of gram-negative bacteria and may indicate the presence of other bacterial products as well. It might be useful to list the level of endotoxins, phage, and other contaminants on each vaccine lot to facilitate studies on any side effects of these contaminants. Selection of vaccine lots with the least endotoxin might reduce some of the adverse effects of vaccinations. PMID:727776

  8. Lipopolysaccaride Detoxification by Endotoxin Neutralizing Protein

    Science.gov (United States)

    1993-08-01

    hemolymph , in induced gross organ lesions, notably in the liver, man. maian serum, antibodies are one of the major pro - spleen, gut-associated lymphoid...derangements that this bacterial toxin can shoe crab , Limuluspolyphemus, detoxifies lipopolysac- induce often end in cardiovascular system collapse or charide...species, including the American horseshoe crab , LPS or LPS/ENP (1:5), only aorta• in the LPSfENP Limulus polyphemus, and the Japanese horseshoe crab

  9. Raphanus sativus L. seeds prevent LPS-stimulated inflammatory response through negative regulation of the p38 MAPK-NF-κB pathway.

    Science.gov (United States)

    Kook, Sung-Ho; Choi, Ki-Choon; Lee, Young-Hoon; Cho, Hyoung-Kwon; Lee, Jeong-Chae

    2014-12-01

    The seeds of Raphanus sativus L. (RSL) have long been used as anti-inflammatory traditional medicine. However, scientific bases for the purported potential of the medicine and the associated mechanisms were barely defined. This study investigated the effects of RSL seeds on lipopolysaccharide (LPS)-stimulated inflammatory responses in vitro and in vivo. Treatment with 100 μg/ml ethyl acetate fraction (REF), which was isolated from water extract of the seeds, significantly inhibited LPS-stimulated production of nitric oxide (P < 0.05), interleukin-6 (P < 0.001), and tumor necrosis factor (TNF)-α (P < 0.001) in RAW264.7 cells. Oral supplementation with 30 mg/kg REF protected mice by 90% against LPS-induced septic death and prevented the increases of serum TNF-α and interferon-γ levels in LPS-injected mice. When REF was divided into four sub-fractions (REF-F1-F4), REF-F3 showed the greatest activity to suppress LPS-stimulated production of inflammatory mediators. We subsequently isolated an active fraction from the REF-F3 and identified sinapic acid as the main constituent. The addition of 50 μg/ml active fraction markedly inhibited LPS-stimulated production of inflammatory mediators by suppressing p38 MAPK and nuclear factor-κB activation. Furthermore, supplementation with the active fraction (10 mg/kg) improved the survival rate of LPS-injected mice by 80% of the untreated control. Additional experiments revealed that sinapic acid was the active component responsible for the anti-inflammatory potential of RSL seeds. Collectively, our current results suggest that both RSL seeds and sinapic acid may be attractive materials for treating inflammatory disorders caused by endotoxins.

  10. Phenylbutazone prevents the endotoxin-induced delay in gastric emptying in horses.

    OpenAIRE

    van der Valk, N.; Doherty, T J; Blackford, J T; Abraha, T W; Frazier, D L

    1998-01-01

    The effect of phenylbutazone on gastric emptying in horses was determined by measuring serum concentrations of acetaminophen. Gastric emptying was determined in normal fasted horses (n = 6), horses given endotoxin intravenously (n = 6), horses given intravenous phenylbutazone (n = 6), and horses given intravenous phenylbutazone plus endotoxin (n = 6). The mean time to reach maximum serum acetaminophen concentration (Tmax), the maximum serum concentration (Cmax), and the area under the serum a...

  11. Allergen specific responses in cord and adult blood are differentially modulated in the presence of endotoxins

    DEFF Research Database (Denmark)

    Eiwegger, T.; Mayer, E.; Pedersen, Susanne Brix

    2008-01-01

    Background Endotoxins are common contaminants in allergen preparations and affect antigen-specific cellular responses. Distinct effects of endotoxin on cells in human umbilical cord and adult blood are poorly defined. Objectives To examine the effect of endotoxins in allergen preparations...... on cellular responses in human cord and peripheral blood (PB). Methods The endotoxin content in beta lactoglobulin (BLG), the peanut allergen Ara h 1 and the major birch pollen allergen Bet v 1 was assessed. Proliferation and cytokine response of mononuclear cells towards contaminated and lipopolysaccharide....... Results The proliferative response of cord blood (CB)-derived mononuclear cells towards allergen-preparations at day 3 was related to the level of LPS contamination. At day 7, proliferation was also detected in the absence of endotoxin. Cytokine production in CB was strongly affected by the content...

  12. Magnetocatalytic Graphene Quantum Dots Janus Micromotors for Bacterial Endotoxin Detection.

    Science.gov (United States)

    Jurado-Sánchez, Beatriz; Pacheco, Marta; Rojo, Jaime; Escarpa, Alberto

    2017-06-06

    Magnetocatalytic hybrid Janus micromotors encapsulating phenylboronic acid (PABA) modified graphene quantum dots (GQDs) are described herein as ultrafast sensors for the detection of deadly bacteria endotoxins. A bottom-up approach was adopted to synthesize an oil-in-water emulsion containing the GQDs along with a high loading of platinum and iron oxide nanoparticles on one side of the Janus micromotor body. The two different "active regions" enable highly efficient propulsion in the presence of hydrogen peroxide or magnetic actuation without the addition of a chemical fuel. Fluorescence quenching was observed upon the interaction of GQDs with the target endotoxin (LPS), whereby the PABA tags acted as highly specific recognition receptors of the LPS core polysaccharide region. Such adaptive hybrid operation and highly specific detection hold considerable promise for diverse clinical, agrofood, and biological applications and integration in future lab-on-chip technology. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Endotoxin recovery using limulus amebocyte lysate (LAL) assay.

    Science.gov (United States)

    Bolden, Jay S; Warburton, Rob E; Phelan, Robert; Murphy, Marie; Smith, Kelly R; De Felippis, Michael R; Chen, Dayue

    2016-09-01

    A phenomenon initially reported by Chen and Vinther in 2013 [1], and now commonly referred to as low endotoxin recovery (LER), has prompted the Food and Drug Administration (FDA) to request specific data demonstrating the capability of the LAL BET method (i.e., USP ) to recover endotoxin from spiked samples over time. The results of these spike/hold recovery studies are expected to be included in the Biologics License Applications (BLA) for review by the Center for Drug Evaluation and Research (CDER) Hughes (2014) and Hughes et al. (2015) [2,3]. Such studies involve spiking a known amount of a surrogate endotoxin, such as purified lipopolysaccharide (LPS), into undiluted biological products and then testing at different time points to determine the recovery over time. We report here the experience and learning gained from conducting spike/hold recovery studies for a monoclonal antibody (Mab) product. Results from initial hold studies spiked with purified LPS showed rapid loss of endotoxin activity in the drug substance (DS) and significant batch-to-batch variation in the drug product (DP). After careful review and examination of the experimental details, it was determined that the study design and execution differed from the routine batch release USP BET method with regard to mixing time and sampling scheme. The hold study design was subsequently revised so that the mixing time and sampling were the same as the verified USP BET method used for routine batch release testing. The spike/hold recovery studies were repeated and the results demonstrated that LPS could be consistently recovered over time. These findings highlight the importance of carefully controlling sample preparation procedures in a spike/hold recovery study in order to demonstrate the suitability of using the LAL BET method for endotoxin detection. Copyright © 2016 International Alliance for Biological Standardization. Published by Elsevier Ltd. All rights reserved.

  14. Endotoxins and inflammation in hemodialysis patients.

    Science.gov (United States)

    El-Koraie, Ahmed F; Naga, Yasmine S; Saaran, Amina M; Farahat, Nahla G; Hazzah, Walaa A

    2013-07-01

    Long-term endotoxin challenge may promote frequent complications in dialysis patients, namely malnutrition, chronic inflammation, and atherosclerosis, which are recognized as the so-called MIA syndrome. Circulating soluble vascular cell adhesion molecule-1 (sVCAM-1) levels may be used to determine the stage of atherosclerosis. This study aimed to assess endotoxin level in hemodialysis (HD) patients and its role in inducing inflammation. The study was conducted on 50 HD patients, chosen from four dialysis centers in Alexandria. Serum blood samples were collected for the determination of albumin and C-reactive protein (CRP), and whole blood samples were used for the measurement of hemoglobin level. A heparinized whole blood sample was taken postdialysis for endotoxin assay by limulus amebocyte lysate test, and in addition to sVCAM-1 was estimated using enzyme-linked immunosorbent assay. The mean endotoxin level was 76.30 pg/mL;80% exhibited values higher than 60 pg/mL. Half the studied patients had CRP values that exceeded the upper limit of the laboratory reference range (endotoxin and CRP levels (r = 0.47, P = 0.001). The mean pre-HD level of VCAM was 1851.00 ng/mL, while the mean post-HD level was 2829.00 ng/mL with statistically significant correlation (r = 0.354, P = 0.012) and it also correlated significantly with endotoxin as well as CRP levels. Endotoxemia may play an important role in the aggravation of endothelial dysfunction in HD patients as indicated by the post-HD rise in sVCAM-1. © 2012 The Authors. Hemodialysis International © 2012 International Society for Hemodialysis.

  15. Extraction of cyanobacterial endotoxin.

    Science.gov (United States)

    Papageorgiou, John; Linke, Thomas A; Kapralos, Con; Nicholson, Brenton C; Steffensen, Dennis A

    2004-02-01

    To simplify our efforts in acquiring toxicological information on endotoxins produced by cyanobacteria, a method development study was undertaken to identify relatively hazard-free and efficient procedures for their extraction. One article sourced and two novel methods were evaluated for their ability to extract lipopolysaccharides (LPSs) or endotoxins from cyanobacteria. The Limulus polyphemus amoebocyte lysate (LAL) assay was employed to compare the performance of a novel method utilizing a 1-butanol-water (HBW) solvent system to that of Westphal's (1965) phenol-water system (HPW) for the extraction of endotoxin from various cyanobacteria. The traditional HPW method extracted from 3- to 12-fold more endotoxin from six different cyanobacterial blooms and culture materials than did the novel HBW method. In direct contrast, the novel HBW method extracted ninefold more endotoxin from a non-microcystin producing Microcystis aeruginosa culture as compared to the HPW method. A solvent system utilizing N,N'-dimethylformamide-water (HDW) was compared to both the HPW and HBW methods for the extraction of endotoxin from natural samples of Anabaena circinalis, Microcystis flos-aquae, and a 1:1 mixture of Microcystis aeruginosa/Microcystisflos-aquae. The LAL activities of these extracts showed that the novel HDW method extracted two- and threefold more endotoxin from the Anabaena sample that did the HBW and HPW methods, respectively. The HDW method also extracted approximately 1.5-fold more endotoxin from the Microcystis flos-aquae sample as compared to both the HBW and HPW methods. On the other hand, the HBW method extracted 2- and 14-fold more endotoxin from the Microcystis flos-aquae/Microcystis aeruginosa mixture than did the HPW and HDW methods, respectively. Results of this study demonstrate that significant disparities exist between the physicochemical properties of the cell wall constituents not only of different cyanobacterial species but also of different strains of

  16. Peptide-assembled graphene oxide as fluorescent turn-on sensor for ultrasensitive Lipopolysaccharide (Endotoxin detection

    Directory of Open Access Journals (Sweden)

    Seng Koon Lim

    2014-06-01

    Full Text Available Introduction: Lipopolysaccharide (LPS, or endotoxin, a major component in the outer cell membrane of Gram-negative bacteria is a very powerful and toxic inflammatory stimulator, resulting in sepsis or septic shock, a significant medical problem affecting about 700 000 patients and causing 250 000 casualties annually in the United States itself. The detection of LPS is highly importance. However, the currently used enzymatic limulus amebocyte lysate assay is highly susceptible to changes in temperature and pH, interference factors, and requires cumbersome sample preparation. A more cost-effective, sensitive and robust detection method is needed. Objective: To design and develop biosensor for LPS detection by assembling a LPS-binding peptide (as LPS receptor with graphene oxide (GO, as fluorescence quencher. Methods: GO was synthesized using a modified Hummer’s method. A synthetic LPS-binding peptide was designed, fluorescent labelled, and assembled with GO in PBS buffer solution. The fluorescence recovery of the peptide-GO was measured upon addition of LPS from Gram negative bacteria: E. coli, K. pneumoniae, Samonella Thyphosa, P. aeruginosa, as well as living pathogenic bacteria. Specificity tests were conducted with various biological molecules to evaluate the sensing performance. Results & Discussion: Specific binding of LPS with peptide release the peptides from GO, resulting in fluorescence recovery, allowing ultrasensitive detection of LPS with the limit of detection of 130 pM, the most sensitive synthetic LPS sensors to-date. The LPS sensor is highly selective to LPS than other biological species. Conclusion: We developed a peptide-GO assembled fluorescence sensor for ultrasensitive and specific LPS/endotoxin detection. This is the most sensitive synthetic LPS sensor reported in the world.

  17. Phenylbutazone prevents the endotoxin-induced delay in gastric emptying in horses.

    Science.gov (United States)

    Valk, N; Doherty, T J; Blackford, J T; Abraha, T W; Frazier, D L

    1998-01-01

    The effect of phenylbutazone on gastric emptying in horses was determined by measuring serum concentrations of acetaminophen. Gastric emptying was determined in normal fasted horses (n = 6), horses given endotoxin intravenously (n = 6), horses given intravenous phenylbutazone (n = 6), and horses given intravenous phenylbutazone plus endotoxin (n = 6). The mean time to reach maximum serum acetaminophen concentration (Tmax), the maximum serum concentration (Cmax), and the area under the serum acetaminophen concentration versus time curve (AUC) were compared among treatment groups. Phenylbutazone did not alter gastric emptying in normal horses. Endotoxin caused a profound delay in gastric emptying, and pretreatment with phenylbutazone abolished this effect. PMID:9684051

  18. Electronic tongue for simultaneous detection of endotoxins and other contaminants of microbiological origin.

    Science.gov (United States)

    Heras, Jorge Yánez; Pallarola, Diego; Battaglini, Fernando

    2010-07-15

    Endotoxins, also referred to as pyrogens, are lipopolysaccharides (LPS) present in the outer membrane of Gram-negative bacteria, and represent one of the most dangerous microbiological contaminants in water for hemodialysis and intravenous infusion. A method is presented for the simultaneous detection of endotoxins and other bacterial lysis contaminating species in purified water for parenteral formulations. The technique used is electrochemical impedance spectroscopy, with data interpretation using principal component analysis (PCA), cluster analysis (CA), and multivariate discriminant analysis (MDA). Two types of electrode surfaces were modified with LPS recognition agents: (i) a 37 amino acids fragment of a 18 kDa cationic antimicrobial protein (CAP18F) that has LPS binding activity; (ii) the highly selective endotoxin neutralizing protein (ENP). Statistical multivariate analysis of the impedance spectral data allowed the detection of endotoxin at, and below, the threshold pharmaceutical regulatory level. Discrimination of LPS from samples containing proteins, nucleic acids, phospholipids or their mixtures was achieved. These results open a new route to a practical instrumental method capable of detecting and discriminating LPS from other potential pro-inflammatory species of microbiological origin, such as nucleic acids. Copyright (c) 2010 Elsevier B.V. All rights reserved.

  19. Serum Levels of Lipopolysaccharide and 1,3-β-D-Glucan Refer to the Severity in Patients with Crohn’s Disease

    Directory of Open Access Journals (Sweden)

    Yanmin Guo

    2015-01-01

    Full Text Available Objectives. Interactions between the host and gut microbial community contribute to the pathogenesis of Crohn’s disease (CD. In this study, we aimed to detect lipopolysaccharide (LPS and 1,3-β-D-glucan (BG in the sera of CD patients and clarify the potential role in the diagnosis and therapeutic approaches. Materials and Methods. Serum samples were collected from 46 patients with active CD (A-CD, 22 CD patients at remission stage (R-CD, and 20 healthy controls, and the levels of LPS, BG, and TNF in sera were determined by ELISA. Moreover, sixteen patients with A-CD received anti-TNF monoclonal antibody therapy (infliximab, IFX at a dose of 5 mg/kg body weight at weeks 0, 2, and 6, and the levels of LPS and BG were also tested at week 12 after the first intravenous infusion. Results. Serum levels of LPS and BG were found to be markedly increased in A-CD patients compared with R-CD patients and healthy controls (P<0.05. They were also observed to be positively correlated with CDAI, ESR, and SES-CD, respectively (P<0.05. Furthermore, the levels of TNF in sera had a significant correlation with LPS and BG, respectively. The concentrations of LPS and BG were demonstrated to be significantly downregulated in the sera of A-CD patients 12 weeks after IFX treatment (P<0.05, suggesting that blockade of TNF could inhibit bacterial endotoxin absorption, partially through improving intestinal mucosal barrier. Conclusions. Serum levels of LPS and BG are significantly increased in A-CD patients and positively correlated with the severity of the disease. Blockade of intestinal mucosal inflammation with IFX could reduce the levels of LPS and BG in sera. Therefore, this study has shed some light on measurement of serum LPS and BG in the diagnosis and treatment of CD patients.

  20. Fucoidan Extracted from Fucus evanescens Prevents Endotoxin-Induced Damage in a Mouse Model of Endotoxemia

    Directory of Open Access Journals (Sweden)

    Tatyana A. Kuznetsova

    2014-01-01

    Full Text Available An important problem of treating patients with endotoxemia is to find drugs to reduce the negative effects of endotoxin on the organism. We tested fucoidan (sulfated polysaccharide from the brown alga Fucus evanescens as a potential drug in a mouse model of endotoxemia inducted by lipopolysaccharide (LPS. The survival time of mice injected with LPS increased under fucoidan treatment compared with the group of mice injected with LPS only. The preventive administration of fucoidan to mice with endotoxemia resulted in inhibition of increased levels of proinflammatory cytokines (TNFα and IL-6, as well as decreasing of the processes of hypercoagulability. The parenteral or per os administration of fucoidan resulted in decreasing the degree of microcirculatory disorders and secondary dystrophic-destructive changes in parenchymal organs of mice with endotoxemia. Taken together, these results demonstrate that fucoidan prevents endotoxin-induced damage in a mouse model of endotoxemia and increases the mice’s resistance to LPS.

  1. Fucoidan extracted from Fucus evanescens prevents endotoxin-induced damage in a mouse model of endotoxemia.

    Science.gov (United States)

    Kuznetsova, Tatyana A; Besednova, Natalya N; Somova, Larisa M; Plekhova, Natalya G

    2014-01-31

    An important problem of treating patients with endotoxemia is to find drugs to reduce the negative effects of endotoxin on the organism. We tested fucoidan (sulfated polysaccharide) from the brown alga Fucus evanescens as a potential drug in a mouse model of endotoxemia inducted by lipopolysaccharide (LPS). The survival time of mice injected with LPS increased under fucoidan treatment compared with the group of mice injected with LPS only. The preventive administration of fucoidan to mice with endotoxemia resulted in inhibition of increased levels of proinflammatory cytokines (TNFα and IL-6), as well as decreasing of the processes of hypercoagulability. The parenteral or per os administration of fucoidan resulted in decreasing the degree of microcirculatory disorders and secondary dystrophic-destructive changes in parenchymal organs of mice with endotoxemia. Taken together, these results demonstrate that fucoidan prevents endotoxin-induced damage in a mouse model of endotoxemia and increases the mice's resistance to LPS.

  2. Influence of the Ward colon tumor on the host response to endotoxin.

    Science.gov (United States)

    Grossie, V B; Mailman, D

    1997-01-01

    Cachexia and a decreased immune function are negative prognostic factors for cancer patients. While the decreased immunity results in a greater susceptibility to bacterial infection, the response of the host to the resulting infection is not clear. The experiments reported here were designed to evaluate the toxicity of endotoxin to rats with a transplantable Ward colon tumor (WCT) and to evaluate the mechanism of the observed increase in lethal toxicity. The lethal toxicity of endotoxin (lipopolysaccharide, LPS) at 5 mg/kg, i.p. was evaluated in the first of two experiments. Rats received LPS and were observed for morbidity and weight loss for a period of 11 days. A second experiment was done to evaluate the effect of LPS on the plasma nitrate/nitrite concentrations and plasma indicators of host tissue dysfunction. LPS was administered as previously described but blood and tissues were collected 5 h after LPS administration. LPS resulted in the death of 1 of 12 nontumor-bearing (NTB) rats and a transient weight loss in the survivors. This same dose of LPS, however, resulted in death for 10 of 12 WCT rats with tumor burdens less than 4% of body weight. The response of WCT rats 5 h after LPS was then compared with that of age-matched NTB rats. Plasma albumin concentrations were not affected by LPS in NTB rats but were significantly decreased in WCT rats. Peripheral blood gases were not consistently affected by LPS in either group. Peripheral blood white cell counts, except monocytes, were significantly decreased by LPS in both groups. Monocyte counts in peripheral blood were further reduced in WCT rats compared with NTB rats receiving LPS. The presence of the WCT significantly enhanced the LPS-associated increase in spleen weight. Liver weights were lower in LPS rats but there was no effect of the presence of WCT. The LPS-associated increase in plasma nitrate/nitrite concentration was enhanced by the WCT. The plasma arginine and citrulline concentrations were altered

  3. Eicosanoids, endotoxins and liver disease

    NARCIS (Netherlands)

    R.J.T. Ouwendijk (Rob)

    1985-01-01

    textabstractEndotoxins are cell wall lipopolysacharides of gram negative bacteria. The gut contains large numbers of bacteria and is generally accepted to be a large reservoir of endotoxins. In the normal state absorbed endotoxins are rapidly removed from the portal blood by especially the

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

    Directory of Open Access Journals (Sweden)

    Ying Sun

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

  5. QMix® irrigant reduces lipopolysacharide (LPS levels in an in vitro model

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    Grasiela Longhi GRÜNDLING

    2015-08-01

    Full Text Available AbstractThe presence of endotoxin inside the root canal has been associated with periapical inflammation, bone resorption and symptomatic conditions.Objectives To determine, in vitro, the effect of QMix® and other three root canal irrigants in reducing the endotoxin content in root canals.Material and Methods Root canals of single-rooted teeth were prepared. Samples were detoxified with Co-60 irradiation and inoculated with E. coli LPS (24 h, at 37°C. After that period, samples were divided into 4 groups, according to the irrigation solution tested: QMix®, 17% EDTA, 2% chlorhexidine solution (CHX, and 3% sodium hypochlorite (NaOCl. LPS quantification was determined by Limulus Amebocyte Lysate (LAL assay. The initial counting of endotoxins for all samples, and the determination of LPS levels in non-contaminated teeth and in contaminated teeth exposed only to non-pyrogenic water, were used as controls.Results QMix® reduced LPS levels, with a median value of 1.11 endotoxins units (EU/mL (p<0.001. NaOCl (25.50 EU/mL, chlorhexidine (44.10 EU/mL and positive control group (26.80 EU/mL samples had similar results. Higher levels were found with EDTA (176.00 EU/mL when compared to positive control (p<0.001. There was no significant difference among EDTA, NaOCl and CHX groups. Negative control group (0.005 EU/mL had statistically significant lower levels of endotoxins when compared to all test groups (p<0.001.Conclusion QMix® decreased LPS levels when compared to the other groups (p<0.001. 3% NaOCl, 2% CHX and 17% EDTA were not able to significantly reduce the root canal endotoxins load.

  6. QMix® irrigant reduces lipopolysacharide (LPS) levels in an in vitro model

    Science.gov (United States)

    GRÜNDLING, Grasiela Longhi; de MELO, Tiago André Fontoura; MONTAGNER, Francisco; SCARPARO, Roberta Kochenborger; VIER-PELISSER, Fabiana Vieira

    2015-01-01

    The presence of endotoxin inside the root canal has been associated with periapical inflammation, bone resorption and symptomatic conditions. Objectives To determine, in vitro, the effect of QMix® and other three root canal irrigants in reducing the endotoxin content in root canals. Material and Methods Root canals of single-rooted teeth were prepared. Samples were detoxified with Co-60 irradiation and inoculated with E. coli LPS (24 h, at 37°C). After that period, samples were divided into 4 groups, according to the irrigation solution tested: QMix®, 17% EDTA, 2% chlorhexidine solution (CHX), and 3% sodium hypochlorite (NaOCl). LPS quantification was determined by Limulus Amebocyte Lysate (LAL) assay. The initial counting of endotoxins for all samples, and the determination of LPS levels in non-contaminated teeth and in contaminated teeth exposed only to non-pyrogenic water, were used as controls. Results QMix® reduced LPS levels, with a median value of 1.11 endotoxins units (EU)/mL (pchlorhexidine (44.10 EU/mL) and positive control group (26.80 EU/mL) samples had similar results. Higher levels were found with EDTA (176.00 EU/mL) when compared to positive control (p<0.001). There was no significant difference among EDTA, NaOCl and CHX groups. Negative control group (0.005 EU/mL) had statistically significant lower levels of endotoxins when compared to all test groups (p<0.001). Conclusion QMix® decreased LPS levels when compared to the other groups (p<0.001). 3% NaOCl, 2% CHX and 17% EDTA were not able to significantly reduce the root canal endotoxins load. PMID:26398517

  7. Endotoxin dosage in sepsis

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    Vincenzo Rondinelli

    2012-03-01

    Full Text Available Introduction. Endotoxin, a component of the cell wall of Gram-negative bacteria is a major contributor to the pathogenesis of septic shock and multiple organ failure (MOF. Its entry into the bloodstream stimulates monocytes/macrophages which once activated produce and release cytokines, nitric oxide and other mediators that induce systemic inflammation, endothelial damage, organ dysfunction, hypotension (shock and MOF.The aim of this study is to evaluate the usefulness of a quantitative test for the dosage of endotoxin to determine the risk of severe Gram-negative sepsis. Materials and methods. In the period January 2009 - June 2011 we performed 897 tests for 765 patients, mostly coming from the emergency room and intensive care, of which 328 (43% women (mean age 53 and 437 (57% male (mean age 49. Fifty-nine patients, no statistically significant difference in sex, were monitored by an average of two determinations of EA.All patients had procalcitonin values significantly altered.The kit used was EAA (Endotoxin Activity Assay Estor Company, Milan, which has three ranges of endotoxin activity (EA: low risk of sepsis if <0.40 units, medium if between 0.40 and 0.59; high if 0.60. Results. 78 out of 765 patients (10% had a low risk, 447 (58% a medium risk and 240 (32% a high risk.The dosage of EA, combined with that of procalcitonin, has allowed a more targeted antibiotic therapy. Six patients in serious clinical conditions were treated by direct hemoperfusion with Toraymyxin, a device comprising a housing containing a fiber polypropylene and polystyrene with surface-bound polymyxin B, an antibiotic that removes bacterial endotoxins from the blood. Conclusions.The test is useful in risk stratification as well as Gram negative sepsis, to set and monitor targeted therapies, also based on the neutralization of endotoxin.

  8. Physicochemical characterization of the endotoxins from Coxiella burnetii strain Priscilla in relation to their bioactivities

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    Hussein Ahmed

    2004-01-01

    Full Text Available Abstract Background Coxiella burnetii is the etiological agent of Q fever found worldwide. The microorganism has like other Gram-negative bacteria a lipopolysaccharide (LPS, endotoxin in its outer membrane, which is important for the pathogenicity of the bacteria. In order to understand the biological activity of LPS, a detailed physico-chemical analysis of LPS is of utmost importace. Results The lipid A moiety of LPS is tetraacylated and has longer (C-16 acyl chains than most other lipid A from enterobacterial strains. The two ester-linked 3-OH fatty acids found in the latter are lacking. The acyl chains of the C. burnetii endotoxins exhibit a broad melting range between 5 and 25°C for LPS and 10 and 40°C for lipid A. The lipid A moiety has a cubic inverted aggregate structure, and the inclination angle of the D-glucosamine disaccharide backbone plane of the lipid A part with respect to the membrane normal is around 40°. Furthermore, the endotoxins readily intercalate into phospholipid liposomes mediated by the lipopolysaccharide-binding protein (LBP. The endotoxin-induced tumor necrosis factor α (TNFα production in human mononuclear cells is one order of magnitude lower than that found for endotoxins from enterobacterial strains, whereas the same activity as in the latter compounds is found in the clotting reaction of the Limulus amebocyte lysate assay. Conclusions Despite a considerably different chemical primary structure of the C. burnetii lipid A in comparison with enterobacterial lipid A, the data can be well understood by applying the previously presented conformational concept of endotoxicity, a conical shape of the lipid A moiety of LPS and a sufficiently high inclination of the sugar backbone plane with respect to the membrane plane. Importantly, the role of the acyl chain fluidity in modulating endotoxicity now becomes more evident.

  9. HYBRID FORMATION BETWEEN BACTERIAL ENDOTOXINS

    Science.gov (United States)

    Rudbach, Jon A.; Milner, Kelsey C.; Ribi, Edgar

    1967-01-01

    When endotoxins extracted from enteric bacteria were mixed in the presence of sodium deoxycholate, and the bile salt was subsequently removed by dialysis or by extraction with ethanol, a new type of endotoxin was formed. The latter material was as biologically active as the original endotoxins and possessed a combination of antigenic determinants that were previously unique to each of the individual endotoxins in the mixture. This hybrid formation between endotoxins was detected by immunodiffusion and radioautography and by quantitative precipitation procedures. PMID:4961269

  10. Selective assay for endotoxin using poly(epsilon-lysine)-immobilized Cellufine and Limulus amoebocyte lysate (LAL).

    Science.gov (United States)

    Sakata, Masayo; Fukuma, Yuriko; Todokoro, Masami; Kunitake, Masashi

    2009-02-15

    We developed a selective endotoxin (lipopolysaccharide; LPS) assay using poly(epsilon-lysine)-immobilized cellulose beads (PL-Cellufine) and Limulus amoebocyte lysate (LAL). First, LPS was selectively adsorbed on the beads in a solution containing various LAL-inhibiting or LAL-enhancing compounds (e.g., amino acids, enzymes) and the LPS adsorbed on the beads was separated from the compounds by centrifugation. Second, the LPS adsorbed on the beads directly reacted with the LAL reagent, and the LPS concentration was determined by a turbidimetric time assay. The accuracy of the adsorption method with PL-Cellufine was high compared with that of a common solution method. Apparent recovery of LPS from compound solution was 88-120%.

  11. Relationships between metabolic changes and clinical signs in pregnant sheep given endotoxin.

    Science.gov (United States)

    Naylor, J M; Kronfeld, D S

    1986-07-01

    Groups of four pregnant ewes were allocated to the following feeding and intravenous endotoxin treatments: fed, Escherichia coli endotoxin (50 micrograms/kg X 75), fed, saline, fasted, E. coli endotoxin (50 micrograms/kg X 75) and fasted, saline. Endotoxin administration resulted in depression, fever, hypoglycemia, hypocalcemia and a reduction in nonesterified fatty acid and ketone body concentrations. Depression correlated best with body temperature (r = 0.76), fasted sheep showed smaller increases in body temperature and were less depressed following endotoxin. Three of eight endotoxin treated sheep died, mortality was not related to rectal temperature but was associated with lactic acidosis. Hypoglycemia was not associated with either death or depression. Fed sheep that were unable to stand had lower serum calcium concentrations than standing sheep.

  12. Removal of endotoxins from bacteriophage preparations by extraction with organic solvents.

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    Bożena Szermer-Olearnik

    Full Text Available Lipopolysaccharide (LPS, endotoxin, pyrogen constitutes a very troubling contaminant of crude phage lysates produced in Gram-negative bacteria. Toxicity of LPS depends on the strong innate immunity response including the cytokines. Therefore, its removal is important for bacteriophage applications. In this paper, we present a procedure for extractive removal of endotoxin from bacteriophage preparations with water immiscible solvents (1-octanol or 1-butanol. During extraction most of the phage lytic activity is retained in the aqueous phase, while endotoxin accumulates in the organic solvent. The levels of endotoxin (expressed as endotoxin units, EU in the aqueous bacteriophage-containing fraction determined by limulus amebocyte lysate or EndoLISA assay were exceptionally low. While the initial endotoxin levels in the crude phage lysates ranged between 10(3 and 10(5 EU/ml the average level after organic extraction remaining in the aqueous fraction was 5.3 EU/ml. These values when related to phage titers decreased from 10(3-10(5 EU/10(9 PFU (plaque forming units down to an average of 2.8 EU/10(9 PFU. The purification procedure is scalable, efficient and applicable to all the bacteriophages tested: T4, HAP1 (E. coli and F8 (P. aeruginosa.

  13. Removal of endotoxins from bacteriophage preparations by extraction with organic solvents.

    Science.gov (United States)

    Szermer-Olearnik, Bożena; Boratyński, Janusz

    2015-01-01

    Lipopolysaccharide (LPS, endotoxin, pyrogen) constitutes a very troubling contaminant of crude phage lysates produced in Gram-negative bacteria. Toxicity of LPS depends on the strong innate immunity response including the cytokines. Therefore, its removal is important for bacteriophage applications. In this paper, we present a procedure for extractive removal of endotoxin from bacteriophage preparations with water immiscible solvents (1-octanol or 1-butanol). During extraction most of the phage lytic activity is retained in the aqueous phase, while endotoxin accumulates in the organic solvent. The levels of endotoxin (expressed as endotoxin units, EU) in the aqueous bacteriophage-containing fraction determined by limulus amebocyte lysate or EndoLISA assay were exceptionally low. While the initial endotoxin levels in the crude phage lysates ranged between 10(3) and 10(5) EU/ml the average level after organic extraction remaining in the aqueous fraction was 5.3 EU/ml. These values when related to phage titers decreased from 10(3)-10(5) EU/10(9) PFU (plaque forming units) down to an average of 2.8 EU/10(9) PFU. The purification procedure is scalable, efficient and applicable to all the bacteriophages tested: T4, HAP1 (E. coli) and F8 (P. aeruginosa).

  14. IgA against gut-derived endotoxins: does it contribute to suppression of hepatic inflammation in alcohol-induced liver disease?

    DEFF Research Database (Denmark)

    Parlesak, Alexandr; Schäfer, C.; Bode, C.

    2002-01-01

    Endotoxins of intestinal origin are supposed to play an important role in the development of alcoholic hepatitis in man. To estimate the role of immunoglobulin response to gut-derived endotoxin in the development of alcohol-induced liver disease, serum levels of IgA and IgG against fecal endotoxin...... production of IgA against endotoxin of intestinal origin may contribute to inactivation of this compound, thereby reducing its damaging effect on the liver....

  15. CR3 (CD11b/CD18) activation of nasal neutrophils: a measure of upper airway endotoxin exposure.

    Science.gov (United States)

    Seth, Romy; Romaschin, Alexander D; Ribeiro, Marcos; Tarlo, Susan M

    2009-11-01

    Inhaled endotoxin (lipopolysaccharide, LPS) initiates an inflammatory response and leads to the expression of CR3 (CD11b/CD18) receptors on polymorphonuclear leukocytes (PMNs). We determined if PMN activation in nasal lavage fluid (NLF) is a possible biomarker of occupational endotoxin exposure. Seven subjects exposed to endotoxin provided NLF samples that were split into three aliquots (negative control--1 M nicotinamide; sham; positive control--11 etag of exogenous LPS) and PMN activation was measured using a chemiluminometer. Differences in mean PMN activation were apparent, negative control: 548 +/- 15.65 RLU 100 microl(-1); sham: 11469 +/- 2582 RLU 100 microl(-1); positive control: 42026 +/- 16659 RLU 100 microl (n = 7; p <0.05). This technique shows promise as a diagnostic method for measuring upper airway LPS exposure.

  16. Membrane adsorber for endotoxin removal

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    Karina Moita de Almeida

    Full Text Available ABSTRACT The surface of flat-sheet nylon membranes was modified using bisoxirane as the spacer and polyvinyl alcohol as the coating polymer. The amino acid histidine was explored as a ligand for endotoxins, aiming at its application for endotoxin removal from aqueous solutions. Characterization of the membrane adsorber, analysis of the depyrogenation procedures and the evaluation of endotoxin removal efficiency in static mode are discussed. Ligand density of the membranes was around 7 mg/g dry membrane, allowing removal of up to 65% of the endotoxins. The performance of the membrane adsorber prepared using nylon coated with polyvinyl alcohol and containing histidine as the ligand proved superior to other membrane adsorbers reported in the literature. The lack of endotoxin adsorption on nylon membranes without histidine confirmed that endotoxin removal was due to the presence of the ligand at the membrane surface. Modified membranes were highly stable, exhibiting a lifespan of approximately thirty months.

  17. The role of phosphodiesterase 3 in endotoxin-induced acute kidney injury

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    Seo Jeong Wook

    2009-06-01

    Full Text Available Abstract Background Acute kidney injury frequently accompanies sepsis. Endotoxin is known to reduce tissue levels of cAMP and low levels of cAMP have been associated with renal injury. We, therefore, hypothesized that endotoxin induced renal injury by activating phosphodiesterase 3 (PDE3 which metabolizes cAMP and that amrinone an inhibitor of PDE3 would prevent the renal injury. Methods Animals were divided into three groups (n = 7/group: 1 Control (0.9% NaCl infusion without LPS; 2 LPS (0.9% NaCl infusion with LPS; 3 Amrinone+LPS (Amrinone infusion with LPS. Either lipopolysaccharide (LPS or vehicle was injected via the jugular vein and the rats followed for 3 hours. We explored the expression of PDE3 isoenzymes and the concentrations of cAMP in the tissue. Results The PDE3B gene but not PDE3A was upregulated in the kidney of LPS group. Immunohistochemistry also showed that PDE3B was expressed in the distal tubule in the controls and LPS caused PDE3B expression in the proximal as well. However, PDE3A was not expressed in the kidney either in the control or LPS treated groups. Tissue level of cAMP was decreased after LPS and was associated with an increase in blood urea nitrogen, creatinine, ultrastructural proximal tubular changes, and expression of inducible nitric oxide synthase (iNOS in the endotoxemic kidney. In septic animals the phosphodiesterase 3 inhibitor, amrinone, preserved the tissue cAMP level, renal structural changes, and attenuated the increased blood urea nitrogen, creatinine, and iNOS expression in the kidney. Conclusion These findings suggest a significant role for PDE3B as an important mediator of LPS-induced acute kidney injury.

  18. The SLE Transcriptome Exhibits Evidence of Chronic Endotoxin Exposure and Has Widespread Dysregulation of Non-Coding and Coding RNAs

    Science.gov (United States)

    Shi, Lihua; Zhang, Zhe; Yu, Angela M.; Wang, Wei; Wei, Zhi; Akhter, Ehtisham; Maurer, Kelly; Reis, Patrícia Costa; Song, Li; Petri, Michelle; Sullivan, Kathleen E.

    2014-01-01

    Background Gene expression studies of peripheral blood mononuclear cells from patients with systemic lupus erythematosus (SLE) have demonstrated a type I interferon signature and increased expression of inflammatory cytokine genes. Studies of patients with Aicardi Goutières syndrome, commonly cited as a single gene model for SLE, have suggested that accumulation of non-coding RNAs may drive some of the pathologic gene expression, however, no RNA sequencing studies of SLE patients have been performed. This study was designed to define altered expression of coding and non-coding RNAs and to detect globally altered RNA processing in SLE. Methods Purified monocytes from eight healthy age/gender matched controls and nine SLE patients (with low-moderate disease activity and lack of biologic drug use or immune suppressive treatment) were studied using RNA-seq. Quantitative RT-PCR was used to validate findings. Serum levels of endotoxin were measured by ELISA. Results We found that SLE patients had diminished expression of most endogenous retroviruses and small nucleolar RNAs, but exhibited increased expression of pri-miRNAs. Splicing patterns and polyadenylation were significantly altered. In addition, SLE monocytes expressed novel transcripts, an effect that was replicated by LPS treatment of control monocytes. We further identified increased circulating endotoxin in SLE patients. Conclusions Monocytes from SLE patients exhibit globally dysregulated gene expression. The transcriptome is not simply altered by the transcriptional activation of a set of genes, but is qualitatively different in SLE. The identification of novel loci, inducible by LPS, suggests that chronic microbial translocation could contribute to the immunologic dysregulation in SLE, a new potential disease mechanism. PMID:24796678

  19. [Effect of endotoxin pretreatment-induced glycogen synthase kinase-3 inhibition on glycogen metabolism in rat liver and the mechanism].

    Science.gov (United States)

    Chen, Xiaole; Gong, Jianping; Xu, Faliang

    2014-02-01

    To investigate the changes in the functional activity of glycogen synthase kinase-3 (GSK-3) in the hepatic tissue after endotoxin (lipopolysaccharide, LPS) tolerance and explore the effects of LPS-induced GSK-3 inhibition on glycogen metabolism in the liver. Male SD rats were randomly divided into normal control, endotoxin pretreatment and GSK-3 inhibitor (lithium chloride) groups with corresponding pretreatments prior to a large dose of LPS challenge (10 mg/kg) to induce liver injury. Glycogen deposition and content in the hepatic tissue was detected using periodic acid-Schiff (PAS) staining and a glycogen quantification kit, respectively. Western blotting was performed for semi-quantitative analysis of protein level and inhibitory phosphorylation of GSK-3, and a Coomassie brilliant blue G-250-based colorimetric assay was used to detect calpain activity in the liver. Glycogen content in the liver decreased significantly after LPS challenge in all the 3 groups (P0.05). Both LPS and lithium chloride pretreatments caused a significant increase of liver glycogen content (P0.05). Large-dose LPS challenge significantly increased the activity of calpain in the liver tissue (P0.05). Endotoxin pretreatment induces inhibitory phosphorylation of GSK-3β and partial cleavage of GSK-3α and promotes the deposition of liver glycogen but does not affect the activity of calpain, which may contribute to an increased glycogen reserve for energy supply in the event of large-dose LPS challenge.

  20. Photoactivated polycationic bioactive chitosan nanoparticles inactivate bacterial endotoxins.

    Science.gov (United States)

    Shrestha, Annie; Cordova, Martha; Kishen, Anil

    2015-05-01

    The current root canal disinfection protocols fail to markedly inactivate bacterial endotoxins from infected root dentin. This study aimed to evaluate the ability of antibacterial photodynamic therapy with chitosan-conjugated rose bengal nanoparticles (CSRBnps) to selectively inactivate endotoxins/lipopolysaccharides (LPSs). Antimicrobial agents such as calcium hydroxide (Ca[OH]2), chitosan nanoparticles (CSnps), CSRBnps, and methylene blue (MB) were assessed for their ability to neutralize LPSs obtained from Pseudomonas aeruginosa in a time-dependent interaction with/without photoactivation (20 and 40 J/cm(2)). The inflammatory potential of the treated/untreated LPSs was assessed on macrophage cells (RAW 267.4) using nitric oxide- and enzyme-linked immunosorbent assay (tumor necrosis factor α and interleukin-6 expression)-based analysis. These antimicrobials were tested directly on macrophage cells for cytotoxicity using the mitochondrial activity assay and light microscopy. The data were analyzed using 1-way analysis of variance and the Tukey test. CSnps were least effective in LPS inactivation. Interluekin-6 expression was reduced only with CSRBnp treatment. CSnps and CSRBnps were completely nontoxic, and MB showed slight toxicity to macrophage cells. Ca(OH)2 was highly cytotoxic (P endotoxins and the subsequent reduction of all tested inflammatory markers from activated macrophages. Antimicrobial CSRBnps in combination with photodynamic therapy showed the potential to effectively inactivate bacterial endotoxins. Copyright © 2015 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  1. Regulation of Lipolysis and Adipose Tissue Signaling during Acute Endotoxin-Induced Inflammation: A Human Randomized Crossover Trial.

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    Nikolaj Rittig

    Full Text Available Lipolysis is accelerated during the acute phase of inflammation, a process being regulated by pro-inflammatory cytokines (e.g. TNF-α, stress-hormones, and insulin. The intracellular mechanisms remain elusive and we therefore measured pro- and anti-lipolytic signaling pathways in adipocytes after in vivo endotoxin exposure.Eight healthy, lean, male subjects were investigated using a randomized cross over trial with two interventions: i bolus injection of saline (Placebo and ii bolus injection of lipopolysaccharide endotoxin (LPS. A 3H-palmitate tracer was used to measure palmitate rate of appearance (Rapalmitate and indirect calorimetry was performed to measure energy expenditures and lipid oxidation rates. A subcutaneous abdominal fat biopsy was obtained during both interventions and subjected to western blotting and qPCR quantifications.LPS caused a mean increase in serum free fatty acids (FFA concentrations of 90% (CI-95%: 37-142, p = 0.005, a median increase in Rapalmitate of 117% (CI-95%: 77-166, p<0.001, a mean increase in lipid oxidation of 49% (CI-95%: 1-96, p = 0.047, and a median increase in energy expenditure of 28% (CI-95%: 16-42, p = 0.001 compared with Placebo. These effects were associated with increased phosphorylation of hormone sensitive lipase (pHSL at ser650 in adipose tissue (p = 0.03, a trend towards elevated pHSL at ser552 (p = 0.09 and cAMP-dependent protein kinase A (PKA phosphorylation of perilipin 1 (PLIN1 (p = 0.09. Phosphatase and tensin homolog (PTEN also tended to increase (p = 0.08 while phosphorylation of Akt at Thr308 tended to decrease (p = 0.09 during LPS compared with Placebo. There was no difference between protein or mRNA expression of ATGL, G0S2, and CGI-58.LPS stimulated lipolysis in adipose tissue and is associated with increased pHSL and signs of increased PLIN1 phosphorylation combined with a trend toward decreased insulin signaling. The combination of these mechanisms appear to be the driving forces

  2. MALP-2, a Mycoplasma lipopeptide with classical endotoxic properties: end of an era of LPS monopoly?

    Science.gov (United States)

    Galanos, C; Gumenscheimer, M; Mühlradt, P; Jirillo, E; Freudenberg, M

    2000-01-01

    Although some activities of LPS are shared by other bacterial components, for half a century LPS has been regarded as unique in displaying many pathophysiological activities. Here we report on a synthetic lipopeptide, MALP-2 from Mycoplasma fermentans, which expresses potent endotoxin-like activity and whose lethal toxicity is comparable to that of LPS. With the exception of the Limulus lysate gelation test, in which MALP-2 was approximately 1000-fold less active than LPS, the synthetic lipopeptide induced all activities tested for, and in most cases to an extent comparable to that of LPS. Unlike LPS, the biological activities of MALP-2 were expressed both in LPS-responder and in LPS-non-responder mice (BALB/c/l, C57BL10/ScCr), indicating that MALP-2 signaling, unlike that of LPS, is not transduced via the Toll-like receptor (Tlr) 4 protein.MALP-2 expressed no toxicity in normal or sensitized Tlr2 knockout (Tlr2(-/-)) mice indicating that its toxic activity is induced via Tlr2 signaling. The phenomenology of the lethal shock induced by MALP-2 in normal or sensitized mice, i.e. the kinetics of its development and symptoms of illness exhibited by the treated animals, was very reminiscent of the lethal shock induced by LPS.

  3. LYATK1 potently inhibits LPS-mediated pro-inflammatory response

    Energy Technology Data Exchange (ETDEWEB)

    Xi, Feng [Department of Intensive Care Unit, Taixing People" ' s Hospital, Taixing, Jiangsu Province, 225400 (China); Liu, Yuan [Department of Ophthalmology, Nanjing First Hospital, Nanjing Medical University, Nanjing (China); Wang, Xiujuan; Kong, Wei [Department of Intensive Care Unit, Taixing People" ' s Hospital, Taixing, Jiangsu Province, 225400 (China); Zhao, Feng, E-mail: taixingzhaofeng163@163.com [Department of Intensive Care Unit, Taixing People" ' s Hospital, Taixing, Jiangsu Province, 225400 (China)

    2016-01-29

    Lipopolysaccharide (LPS)-primed monocytes/macrophages produce pro-inflammatory cytokines, which could lead to endotoxin shock. TGF-β-activated kinase1 (TAK1) activation is involved in the process. In the current study, we studied the potential effect of a selective TAK1 inhibitor, LYTAK1, on LPS-stimulated response both in vitro and in vivo. We demonstrated that LYTAK1 inhibited LPS-induced mRNA expression and production of several pro-inflammatory cytokines [interleukin 1β (IL-1β), tumor necrosis factor-α (TNFα) and interleukin-6 (IL-6)] in RAW 264.7 macrophages. LYTAK1's activity was almost nullified with TAK1 shRNA-knockdown. Meanwhile, in both primary mouse bone marrow derived macrophages (BMDMs) and human peripheral blood mononuclear cells (PBMCs), LPS-induced pro-inflammatory cytokine production was again attenuated with LYTAK1 co-treatment. Molecularly, LYTAK1 dramatically inhibited LPS-induced TAK1-nuclear factor kappa B (NFκB) and mitogen-activated protein kinase (Erk, Jnk and p38) activation in RAW 264.7 cells, mouse BMDMs and human PBMCs. In vivo, oral administration of LYTAK1 inhibited LPS-induced activation of TAK1-NFκB-p38 in ex-vivo cultured PBMCs, and cytokine production and endotoxin shock in mice. Together, these results demonstrate that LYTAK1 inhibits LPS-induced production of several pro-inflammatory cytokines and endotoxin shock probably through blocking TAK1-regulated signalings. - Highlights: • LYTAK1 inhibits LPS-induced pro-inflammatory cytokine production in RAW 264.7 cells. • The effect by LYTAK1 is more potent than other known TAK1 inhibitors. • LYTAK1 inhibits LPS-induced cytokine production in primary macrophages/monocytes. • LYTAK1 inhibits LPS-induced TAK1-NFκB and MAPK activation in macrophages/monocytes. • LYTAK1 gavage inhibits LPS-induced endotoxin shock and cytokine production in mice.

  4. Adsorption of endotoxins on Ca2+ -iminodiacetic acid by metal ion affinity chromatography.

    Science.gov (United States)

    Lopes, André Moreni; Romeu, Jorge Sánchez; Meireles, Rolando Páez; Perera, Gabriel Marquez; Morales, Rolando Perdomo; Pessoa, Adalberto; Cárdenas, Lourdes Zumalacárregui

    2012-11-01

    Endotoxins (also known as lipopolysaccharides (LPS)) are undesirable by-products of recombinant proteins, purified from Escherichia coli. LPS can be considered stable under a wide range of temperature and pH, making their removal one of the most difficult tasks in downstream processes during protein purification. The inherent toxicity of LPS makes their removal an important step for the application of these proteins in several biological assays and for a safe parenteral administration. Immobilized metal affinity chromatography (IMAC) enables the affinity interactions between the metal ions (immobilized on the support through the chelating compound) and the target molecules, thus enabling high-efficiency separation of the target molecules from other components present in a mixture. Affinity chromatography is applied with Ca2+ -iminodiacetic acid (IDA) to remove most of the LPS contaminants from the end product (more than 90%). In this study, the adsorption of LPS on an IDA-Ca2+ was investigated. The adsorption Freundlich isotherm of LPS-IDA-Ca2+ provides a theoretical basis for LPS removal. It was found that LPS is bound mainly by interactions between the phosphate group in LPS and Ca2+ ligands on the beads. The factors such as pH (4.0 or 5.5) and ionic strength (1.0 mol/L) are essential to obtain effective removal of LPS for contaminant levels between endotoxin' concentration values less than 100 EU/mL and 100 000 EU/mL. This new protocol represents a substantial advantage in time, effort, and production costs.

  5. Synchronous recruitment of epigenetic modifiers to endotoxin synergistically activated Tnf-α gene in acute kidney injury.

    Directory of Open Access Journals (Sweden)

    Karol Bomsztyk

    Full Text Available As a consequence of acute kidney injury (AKI, proximal tubular cells hyperrespond to endotoxin (lipopolysaccharide, LPS by exaggerated renal Tnf-α Production. This LPS hyperresponsiveness is transcriptionally mediated. The epigenetic pathways that control these responses are unknown.We applied multiplex chromatin immunoprecipitation platform (Matrix ChIP to explore epigenetic pathways that underlie endotoxin hyperresponsiveness in the setting of preceding unilateral renal ischemia/reperfusion (I/R in mouse AKI model. Endotoxin exposure after I/R resulted in enhanced transcription, manifested by hyperresponsive recruitment of RNA polymerase II (Pol II at the Tnf-α gene. At this locus, LPS but not I/R increased levels of Pol II C-terminal domain (CTD phosho-serine2 &5 and induced dephosphorylation of the transcription-repressive histone H4 phospho-serine-1. In contrast, I/R but not LPS increased the transcription-permissive histone phosphorylation (H3 phospho-serine-10, H3.3 phospho-serine-31 at the Tnf-α gene. In agreement with these observations, I/R but not LPS increased activity of cognate kinases (Erk1/2, Msk1/2 and Aurora A at the Tnf-α locus. Cross-talk of histone phosphorylation and acetylation synergize to active gene expression. I/R and LPS increased histone acetylation. (H3K9/14Ac, H4K5/8/12/16Ac, H2KA5Ac, H2BK4/7Ac. Levels of some histone acetyltransferases at this gene (PCAF and MOF were increased by I/R but not by LPS, while others were induced by either I/R or LPS and exhibited endotoxin hyperresponsive patterns (GCN5, CBP and p300. The adaptor protein 14-3-3 couples histone phosphorylation with acetylation, and tethers chromatin modifiers/transcription elongation factors to target genes. Both I/R and LPS increased levels of 14-3-3 and several chromatin/transcription modifiers (BRD4, BRG1, HP-1γ and IKKα at the Tnf-α gene, all exhibiting endotoxin hyperresponsive recruitment patterns similar to Pol II.Our results suggest that

  6. Thalidomide protects mice against LPS-induced shock

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    Moreira A.L.

    1997-01-01

    Full Text Available Thalidomide has been shown to selectively inhibit TNF-a production in vitro by lipopolysaccharide (LPS-stimulated monocytes. TNF-a has been shown to play a pivotal role in the pathophysiology of endotoxic shock. Using a mouse model of LPS-induced shock, we investigated the effects of thalidomide on the production of TNF-a and other cytokines and on animal survival. After injection of 100-350 µg LPS into mice, cytokines including TNF-a, IL-6, IL-10, IL-1ß, GM-CSF and IFN-g were measured in the serum. Administration of 200 mg/kg thalidomide to mice before LPS challenge modified the profile of LPS-induced cytokine secretion. Serum TNF-a levels were reduced by 93%, in a dose-dependent manner, and TNF-a mRNA expression in the spleens of mice was reduced by 70%. Serum IL-6 levels were also inhibited by 50%. Thalidomide induced a two-fold increase in serum IL-10 levels. Thalidomide treatment did not interfere with the production of GM-CSF, IL-1ß or IFN-g. The LD50 of LPS in this model was increased by thalidomide pre-treatment from 150 µg to 300 µg in 72 h. Thus, at otherwise lethal doses of LPS, thalidomide treatment was found to protect animals from death

  7. The Potential Role of an Endotoxin Tolerance-Like Mechanism for the Anti-Inflammatory Effect of Spirulina platensis Organic Extract in Macrophages.

    Science.gov (United States)

    Pham, Tho X; Park, Young-Ki; Bae, Minkyung; Lee, Ji-Young

    2017-03-01

    Endotoxin tolerance is a phenomenon where exposure of innate immune cells to lipopolysaccharide (LPS) induces a refractory state to subsequent endotoxin exposures. The goal of this study was to investigate if Spirulina platensis organic extract (SPE) induces an endotoxin tolerance-like state. We used splenocytes and peritoneal macrophages from C57BL/6J mice fed a high-fat/high-sucrose (HF/HS) control or a HF/HS diet containing 0.25% (w/w) SPE for 16 weeks for ex vivo LPS stimulation and endotoxin-tolerant (ET) macrophages to evaluate the effects of SPE on endotoxin tolerance. Cells from SPE-fed mice displayed significantly less expression of proinflammatory genes than those from control mice. ET macrophages were produced in vitro by incubating RAW 264.7 macrophages with low-dose LPS to determine the energy phenotype of naive, SPE-treated, and ET macrophages. Compared to naive macrophages exposed to a high-dose LPS (100 ng/mL) for the first time, ET macrophages showed significantly less proinflammatory gene expression after LPS stimulation, which was also observed with SPE treatment. Consistently, nuclear translocation of p65 was markedly reduced in both ET- and SPE-treated macrophages on LPS stimulation with increase in nuclear protein levels of p50 and B cell lymphoma 3-encoded protein. In conclusion, the anti-inflammatory effect of SPE is at least partly attributable to the induction of an endotoxin tolerance-like state in macrophages, which shares common characteristics of macrophage endotoxin tolerance.

  8. Critical role of P-selectin-dependent leukocyte recruitment in endotoxin-induced intestinal barrier dysfunction in mice.

    OpenAIRE

    Mangell, Peter; Röme, Andrada; Wang, Yusheng; Schramm, R; Jeppsson, Bengt; Thorlacius, Henrik

    2007-01-01

    Objective: To define the importance of leukocyte recruitment in endotoxin-induced gut permeability. Materials and methods: 31 male C57BL/6 mice were challenged with lipopolysaccharide (LPS). Ileal permeability was measured in Ussing chambers and leukocyte-endothelium interactions studied with intravital fluorescence microscopy after 18 h. Results: LPS caused a clear-cut increase in leukocyte accumulation and intestinal permeability. Immunoneutralisation of P-selectin not only reduced leukocyt...

  9. Endotoxin hitchhiking on polymer nanoparticles

    Science.gov (United States)

    Donnell, Mason L.; Lyon, Andrew J.; Mormile, Melanie R.; Barua, Sutapa

    2016-07-01

    The control of microbial infections is critical for the preparation of biological media including water to prevent lethal septic shock. Sepsis is one of the leading causes of death in the United States. More than half a million patients suffer from sepsis every year. Both gram-positive and gram-negative bacteria are responsible for septic infection by the most common organisms i.e., Escherichia coli and Pseuodomonas aeruginosa. The bacterial cell membrane releases negatively charged endotoxins upon death and enzymatic destruction, which stimulate antigenic response in humans to gram-negative infections. Several methods including distillation, ethylene oxide treatment, filtration and irradiation have been employed to remove endotoxins from contaminated samples, however, the reduction efficiency remains low, and presents a challenge. Polymer nanoparticles can be used to overcome the current inability to effectively sequester endotoxins from water. This process is termed endotoxin hitchhiking. The binding of endotoxin on polymer nanoparticles via electrostatic and hydrophobic interactions offers efficient removal from water. However, the effect of polymer nanoparticles and its surface areas has not been investigated for removal of endotoxins. Poly(ε-caprolactone) (PCL) polymer was tested for its ability to effectively bind and remove endotoxins from water. By employing a simple one-step phase separation technique, we were able to synthesize PCL nanoparticles of 398.3 ± 95.13 nm size and a polydispersity index of 0.2. PCL nanoparticles showed ∼78.8% endotoxin removal efficiency, the equivalent of 3.9 × 105 endotoxin units (EU) per ml. This is 8.34-fold more effective than that reported for commercially available membranes. Transmission electron microscopic images confirmed binding of multiple endotoxins to the nanoparticle surface. The concept of using nanoparticles may be applicable not only to eliminate gram-negative bacteria, but also for any gram

  10. Septic Participation in Cardiogenic Shock: Exposure to Bacterial Endotoxin.

    Science.gov (United States)

    Ramirez, Paula; Villarreal, Esther; Gordon, Monica; Gómez, María Dolores; de Hevia, Luis; Vacacela, Karla; Gisbert, Teresa; Quinzá, Adrian; Ruiz, Jesús; Alonso, Ricardo; Bonastre, Juan; Vila, Jordi

    2017-05-01

    In cardiogenic shock (CS), presence of fever, leukocytosis, relatively low systemic vascular resistances, and high serum procalcitonin levels are quite frequent and recurrently involve the search for an infectious complication. We hypothesized that endotoxin exposure in CS could explain this sepsis-like syndrome. Prospective observational study of consecutive CS patients admitted to our intensive care unit (ICU). Patients were followed during the first 3 days after CS onset. All clinical, hemodynamic, and microbiological data were collected. Inflammatory biomarkers and anti-endotoxin antibodies (IgM EndoCAb) were daily measured. We included 37 consecutive CS patients. Twenty-two patients (60%) had body temperature >38.3°C or 14,000/mm or <4,000/mm. Microbiological study was performed in 30 patients (81%). No infection was diagnosed in the studied patients. All the patients had serum inflammatory biomarkers levels above normal values including procalcitonin (19 patients [51%] had serum procalcitonin above 2 ng/mL). All the patients had IgM EndoCAb below the normal median value; 22 patients (59.5%) had IgM anti-endotoxin value below 10th percentile range for healthy people. Hemodynamic and respiratory stabilization was achieved in 23 patients (62%) and the ICU mortality rate was 38%, only procalcitonin and interleuquin-6 were associated with higher mortality rate. We have detected extremely low titers of IgM EndoCAb in CS suggesting endotoxin exposure. However, only inflammatory biomarkers were related to ICU mortality.

  11. Complex links between dietary lipids, endogenous endotoxins and metabolic inflammation.

    Science.gov (United States)

    Laugerette, Fabienne; Vors, Cécile; Peretti, Noël; Michalski, Marie-Caroline

    2011-01-01

    Metabolic diseases such as obesity are characterized by a subclinical inflammatory state that contributes to the development of insulin resistance and atherosclerosis. Recent reports also indicate that (i) there are alterations of the intestinal microbiota in metabolic diseases and (ii) absorption of endogenous endotoxins (namely lipopolysaccharides, LPS) can occur, particularly during the digestion of lipids. The aim of the present review is to highlight recently gained knowledge regarding the links between high fat diets, lipid digestion, intestinal microbiota and metabolic endotoxemia & inflammation. Copyright © 2010 Elsevier Masson SAS. All rights reserved.

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

    Science.gov (United States)

    Sun, Ying; Li, Hui; Yang, Mi-Fang; Shu, Wei; Sun, Meng-Jun; Xu, Yan

    2012-01-01

    Periodontitis is a bacterially induced chronic inflammatory disease. Exposure of the host to periodontal pathogens and their virulence factors induces a state of hyporesponsiveness to subsequent stimulations, termed endotoxin tolerance. Aging has a profound effect on immune response to bacteria challenge. The aim of this study was to explore the effects of aging on endotoxin tolerance induced by Porphyromonas gingivalis (P. gingivalis) lipopolysaccharide (LPS) and Escherichia coli (E. coli) LPS in murine peritoneal macrophages. We studied the cytokine production (TNF-α and IL-10) and Toll-like receptor 2, 4 (TLR2, 4) gene and protein expressions in peritoneal macrophages from young (2-month-old) and middle-aged (12-month-old) ICR mice following single or repeated P. gingivalis LPS or E. coli LPS stimulation. Pretreatment of peritoneal macrophages with P. gingivalis LPS or E. coli LPS resulted in a reduction in TNF-α production and an increase in IL-10 production upon secondary stimulation (plead to the incontrollable periodontal inflammation in older adults.

  13. Endotoxin-induced inflammation disturbs melatonin secretion in ewe

    Directory of Open Access Journals (Sweden)

    Andrzej Przemysław Herman

    2017-12-01

    Full Text Available Objective The study examined the effect of intravenous administration of bacterial endotoxin—lipopolysaccharide (LPS —on the nocturnal secretion of melatonin and on the expression of enzymes of the melatonin biosynthetic pathway in the pineal gland of ewes, taking into account two different photoperiodic conditions: short-night (SN; n = 12 and long-night (LN; n = 12. Methods In both experiments, animals (n = 12 were randomly divided into two groups: control (n = 6 and LPS-treated (n = 6 one. Two hours after sunset, animals received an injection of LPS or saline. Blood samples were collected starting one hour after sunset and continuing for 3 hours after the treatment. The ewes were euthanized 3 hours after LPS/saline treatment. The concentration of hormones in plasma was assayed by radioimmunoassay. In the pineal gland, the content of serotonin and its metabolite was determined by HPLC; whereas the expression of examined genes and protein was assayed using real-time polymerase chain reaction and Western Blot, respectively. Results Endotoxin administration lowered (p<0.05 levels of circulating melatonin in animals from LN photoperiod only during the first hour after treatment, while in ewes from SN photoperiod only in the third hour after the injection. Inflammation more substantially suppressed biosynthesis of melatonin in ewes from SN photoperiod, which were also characterised by lower (p<0.05 cortisol concentrations after LPS treatment compared with animals from LN photoperiod. In the pineal gland of ewes subjected to SN photoperiod, LPS reduced (p<0.05 serotonin content and the expression of melatonin biosynthetic pathway enzymes, such as tryptophan hydroxylase and arylalkylamine-N-acetyltransferase. Pineal activity may be disturbed by circulating LPS and proinflammatory cytokines because the expression of mRNAs encoding their corresponding receptors was determined in this gland. Conclusion The present study showed that peripheral

  14. Synergism between endotoxin priming and exotoxin challenge in provoking severe vascular leakage in rabbit lungs.

    Science.gov (United States)

    Schütte, H; Rosseau, S; Czymek, R; Ermert, L; Walmrath, D; Krämer, H J; Seeger, W; Grimminger, F

    1997-09-01

    Lipopolysaccharides (LPS) of gram-negative bacteria prime rabbit lungs for enhanced thromboxane-mediated vasoconstriction upon subsequent challenge with the exotoxin Escherichia coli hemolysin (HlyA) (Walmrath et al. J. Exp. Med. 1994;180:1437-1443). We investigated the impact of endotoxin priming and subsequent HlyA challenge on lung vascular permeability while maintaining constancy of capillary pressure. Rabbit lungs were perfused in a pressure-controlled mode in the presence of the thromboxane receptor antagonist BM 13.505, with continuous monitoring of flow. Perfusion for 180 min with 10 ng/ml LPS did not provoke vasoconstriction or alteration of capillary filtration coefficient (Kfc) values. HlyA (0.021 hemolytic units/ml) induced thromboxane release and a transient decrease in perfusion flow in the absence of significant changes in Kfc. Similar results were obtained when LPS and HlyA were coapplied simultaneously. However, when the HlyA challenge was undertaken after 180 min of LPS priming, a manifold increase in Kfc values was noted, with concomitant severe lung edema formation, although capillary pressure remained unchanged. Thus, endotoxin primes the lung vasculature to respond with a severe increase in vascular permeability to a subsequent low-dose application of HlyA. Such synergism between endotoxin priming and exotoxin challenge in provoking lung vascular leakage may contribute to the pathogenesis of respiratory failure in sepsis and severe lung infection.

  15. A central role for the mammalian target of rapamycin in LPS-induced anorexia in mice.

    Science.gov (United States)

    Yue, Yunshuang; Wang, Yi; Li, Dan; Song, Zhigang; Jiao, Hongchao; Lin, Hai

    2015-01-01

    Bacterial lipopolysaccharide (LPS), also known as endotoxin, induces profound anorexia. However, the LPS-provoked pro-inflammatory signaling cascades and the neural mechanisms underlying the development of anorexia are not clear. Mammalian target of rapamycin (mTOR) is a key regulator of metabolism, cell growth, and protein synthesis. This study aimed to determine whether the mTOR pathway is involved in LPS-induced anorexia. Effects of LPS on hypothalamic gene/protein expression in mice were measured by RT-PCR or western blotting analysis. To determine whether inhibition of mTOR signaling could attenuate LPS-induced anorexia, we administered an i.c.v. injection of rapamycin, an mTOR inhibitor, on LPS-treated male mice. In this study, we showed that LPS stimulates the mTOR signaling pathway through the enhanced phosphorylation of mTOR(Ser2448) and p70S6K(Thr389). We also showed that LPS administration increased the phosphorylation of FOXO1(Ser256), the p65 subunit of nuclear factor kappa B (Panorexia by decreasing the phosphorylation of p70S6K(Thr389), FOXO1(Ser256), and FOXO1/3a(Thr) (24) (/) (32). These results suggest promising approaches for the prevention and treatment of LPS-induced anorexia. © 2015 Society for Endocrinology.

  16. Toll-like receptors (TLR) 2 and 4 on human sperm recognize bacterial endotoxins and mediate apoptosis.

    Science.gov (United States)

    Fujita, Youko; Mihara, Toshihiro; Okazaki, Tetsuji; Shitanaka, Manami; Kushino, Rena; Ikeda, Chiaki; Negishi, Hiroaki; Liu, Zhilin; Richards, JoAnne S; Shimada, Masayuki

    2011-10-01

    Bacterial infections of the genital tract are one of the most serious causes of infertility in males. In some human patients with poor semen quality, leukocytospermia has been observed. Because leukocytes express the bacterial-lipopolysaccharide (LPS) responsive Toll-like receptor (TLR) signaling cascade and secrete tumor necrosis factor-α, secreted cytokines comprise one, but probably not the only, class of factors that can impact sperm motility. In this study, we documented that bacterial endotoxins, LPS and peptidoglycan, can be detected in human semen. Furthermore, the addition of endotoxins in the absence of leukocytes directly and significantly reduced the motility and increased the apoptotic rate of both human and mouse sperm and suppressed fertilization by mouse sperm both in vivo and in vitro. The well-known LPS receptor, TLR4, and peptidoglycan receptor, TLR2, were expressed in human and mouse sperm. In Tlr2/4(-/-) double-mutant mice, the negative effects of endotoxins on sperm functions were blocked, suggesting that the bacterial endotoxins mediated activation of TLR-dependent pathways in sperm leading to apoptosis. Sperm can recognize bacterial endotoxins by TLRs present in their membranes. The activated TLRs reduce sperm motility, induce sperm apoptosis and significantly impair the potential for fertilization.

  17. Detection of bacterial endotoxin in food: New planar interdigital sensors based approach

    KAUST Repository

    Abdul Rahman, Mohd Syaifudin

    2013-02-01

    Food poisoning caused by endotoxins or Lipopolysaccharide (LPS) are associated with Gram-negative bacteria. Two major food-borne pathogens, Escherichia coli and Salmonella are examples of Gram-negative bacteria which cause a large number of outbreaks of food poisoning. New types of planar interdigital sensors have been fabricated with different coating materials to assess their response to endotoxins. A carboxyl-functional polymer, APTES (3-Aminopropyltriethoxysilane) and Thionine were chosen to be coated onto FR4 interdigital sensors. The chosen coating materials have carboxylic or amine functional groups, which were optimized to be stable in water. All coated sensors were immobilized with PmB (Polymyxin B) which has specific binding properties to LPS. The sensors were tested with different concentrations of LPS O111:B4, ranging from 0.1 to 1000 μg/ml. Analyses of sensors\\' performance were based on the impedance spectroscopy method. The impedance spectra were modeled using a constant phase-element (CPE) equivalent circuit, and a principal component analysis (PCA) was used for data classification. Sensor coated with APTES has shown better selectivity for LPS detection. The experiments were repeated by coating APTES and immobilizing PmB to a new improve designed of novel interdigital sensors (thin film silicon based sensors). These sensors were observed to have better sensitivity and selectivity to the target biomolecules of LPS. Further experiments were conducted to study the effect of different coating thickness on sensor sensitivity, selectivity and stability. Different food samples contaminated with endotoxin were also tested to verify that the interdigital sensing approach is able to be used for endotoxin detection. © 2012 Elsevier Ltd. All rights reserved.

  18. Evaluation of the ability of LL-37 to neutralise LPS in vitro and ex vivo.

    Science.gov (United States)

    Scott, Aaron; Weldon, Sinéad; Buchanan, Paul J; Schock, Bettina; Ernst, Robert K; McAuley, Danny F; Tunney, Michael M; Irwin, Chris R; Elborn, J Stuart; Taggart, Clifford C

    2011-01-01

    Human cathelicidin LL-37 is a cationic antimicrobial peptide (AMP) which possesses a variety of activities including the ability to neutralise endotoxin. In this study, we investigated the role of LPS neutralisation in mediating LL-37's ability to inhibit Pseudomonas aeruginosa LPS signalling in human monocytic cells. Pre-treatment of monocytes with LL-37 significantly inhibited LPS-induced IL-8 production and the signalling pathway of associated transcription factors such as NF-κB. However, upon removal of LL-37 from the media prior to LPS stimulation, these inhibitory effects were abolished. These findings suggest that the ability of LL-37 to inhibit LPS signalling is largely dependent on extracellular LPS neutralisation. In addition, LL-37 potently inhibited cytokine production induced by LPS extracted from P. aeruginosa isolated from the lungs of cystic fibrosis (CF) patients. In the CF lung, polyanionic molecules such as glycosaminoglycans (GAGs) and DNA bind LL-37 and impact negatively on its antibacterial activity. In order to determine whether such interactions interfere with the LPS neutralising ability of LL-37, the status of LL-37 and its ability to bind LPS in CF sputum were investigated. Overall our findings suggest that in the CF lung, the ability of LL-37 to bind LPS and inhibit LPS-induced IL-8 production is attenuated as a result of binding to DNA and GAGs. However, LL-37 levels and its concomitant LPS-binding activity can be increased with a combination of DNase and GAG lyase (heparinase II) treatment. Overall, these findings suggest that a deficiency in available LL-37 in the CF lung may contribute to greater LPS-induced inflammation during CF lung disease.

  19. Evaluation of the ability of LL-37 to neutralise LPS in vitro and ex vivo.

    Directory of Open Access Journals (Sweden)

    Aaron Scott

    Full Text Available BACKGROUND: Human cathelicidin LL-37 is a cationic antimicrobial peptide (AMP which possesses a variety of activities including the ability to neutralise endotoxin. In this study, we investigated the role of LPS neutralisation in mediating LL-37's ability to inhibit Pseudomonas aeruginosa LPS signalling in human monocytic cells. METHODOLOGY/PRINCIPAL FINDINGS: Pre-treatment of monocytes with LL-37 significantly inhibited LPS-induced IL-8 production and the signalling pathway of associated transcription factors such as NF-κB. However, upon removal of LL-37 from the media prior to LPS stimulation, these inhibitory effects were abolished. These findings suggest that the ability of LL-37 to inhibit LPS signalling is largely dependent on extracellular LPS neutralisation. In addition, LL-37 potently inhibited cytokine production induced by LPS extracted from P. aeruginosa isolated from the lungs of cystic fibrosis (CF patients. In the CF lung, polyanionic molecules such as glycosaminoglycans (GAGs and DNA bind LL-37 and impact negatively on its antibacterial activity. In order to determine whether such interactions interfere with the LPS neutralising ability of LL-37, the status of LL-37 and its ability to bind LPS in CF sputum were investigated. Overall our findings suggest that in the CF lung, the ability of LL-37 to bind LPS and inhibit LPS-induced IL-8 production is attenuated as a result of binding to DNA and GAGs. However, LL-37 levels and its concomitant LPS-binding activity can be increased with a combination of DNase and GAG lyase (heparinase II treatment. CONCLUSIONS/SIGNIFICANCE: Overall, these findings suggest that a deficiency in available LL-37 in the CF lung may contribute to greater LPS-induced inflammation during CF lung disease.

  20. Mononuclear cells in the corneal response to endotoxin

    Energy Technology Data Exchange (ETDEWEB)

    Howes, E.L.; Cruse, V.K.; Kwok, M.T.

    1982-04-01

    A severe keratitis can be produced after the direct injection of bacterial endotoxin, or lipopolysaccharide (LPS), in rabbits. Corneal inflammation can progress to scarring and vascularization within a 2 to 3 week period. Pretreatment with systemic adrenal corticosteroids (triamcinolone) prevents this response. Limbal cellular and vascular events were studied during the first 20 hr after injection of LPS in treated and nontreated rabbits. Perivascular limbal inflammatory cells were counted and limbal vascular permeability was assessed by extravasation of 131I-albumin and 125I-fibrinogen in the cornea. Corticosteroids decreased but did not prevent the early protein extravasation and profoundly altered the inflammatory cell population around blood vessels at the limbus. Mononuclear cells, particularly mononuclear phagocytes, were sharply reduced. It is proposed that these cell types play an important role in the perpetuation and amplification of the inflammatory response in this reaction.

  1. Detection and quantitative evaluation of endotoxin contamination in nanoparticle formulations by LAL-based assays.

    Science.gov (United States)

    Neun, Barry W; Dobrovolskaia, Marina A

    2011-01-01

    Bacterial endotoxin or lipopolysaccharide (LPS) is a membrane component of all Gram-negative bacteria. The administration of products contaminated with bacterial endotoxin can cause fever, shock, and even death. Accordingly, the FDA sets limits on the number of endotoxin units (EU) that may be present in a drug or device product. Limulus amoebocyte lysate (LAL) is the extract from amoebocytes of the horseshoe crab Limulus polyphemus, which reacts with bacterial endotoxin. Detection of the products of this reaction is an effective means of quantifying the EU present in a drug formulation. However, nanoparticles frequently interfere with the reactivity of endotoxin, the LAL reaction, or the detection of the reaction products. This interference can be manifested as either an enhancement or an inhibition, causing a respective overestimation or underestimation of the EU in the sample. Here, we present two methods for the detection and quantification of endotoxin in nanoparticle preparations: one is based on an end-point chromogenic LAL assay, and the second approach is based on measuring the turbidity of the LAL extract.

  2. Dried citrus pulp modulates the physiological and acute phase responses of crossbred heifers to an endotoxin challenge

    Science.gov (United States)

    This study examined the effect of feeding dried citrus pulp (CP) pellets on the physiological and acute phase responses (APR) of newly-received crossbred heifers to an endotoxin (lipopolysaccharide; LPS) challenge. Heifers (n=24; 218.3±2.4 kg) were obtained from commercial sale barns and transported...

  3. Ultrasound body composition traits response to an endotoxin challenge in Brahman heifers supplemented with Omnigen-AF

    Science.gov (United States)

    This study examined the effect of feeding OmniGen-AF (OG; Prince Agri Products) on the body composition traits response of newly-weaned heifers to an endotoxin (lipopolysaccharide; LPS) challenge. Brahman heifers (n=24; 183 ± 5 kg) from the Texas AgriLife Research Center in Overton, TX, were separat...

  4. Sexually dimorphic secretion of cortisol but not catecholamines in response to an endotoxin challenge in beef cattle

    Science.gov (United States)

    This study was designed to determine the effect of endotoxin (lipopolysaccharide; LPS) challenge on secretion of the adrenal stress-related hormones cortisol, epinephrine, and norepinephrine in bull and heifer calves. Brahman calves (n = 12; 269 ± 11.7 kg) were randomly selected from the fall 2007 c...

  5. Chlorogenic acid ameliorates endotoxin-induced liver injury by promoting mitochondrial oxidative phosphorylation

    Energy Technology Data Exchange (ETDEWEB)

    Zhou, Yan [State Key Laboratory of Food Science and Technology and School of Food Science, Nanchang University, Nanchang 330047 (China); College of Food Safety, Guizhou Medical University, Guiyang 550025 (China); Ruan, Zheng, E-mail: ruanzheng@ncu.edu.cn [State Key Laboratory of Food Science and Technology and School of Food Science, Nanchang University, Nanchang 330047 (China); Zhou, Lili; Shu, Xugang [State Key Laboratory of Food Science and Technology and School of Food Science, Nanchang University, Nanchang 330047 (China); Sun, Xiaohong [College of Food Safety, Guizhou Medical University, Guiyang 550025 (China); Mi, Shumei; Yang, Yuhui [State Key Laboratory of Food Science and Technology and School of Food Science, Nanchang University, Nanchang 330047 (China); Yin, Yulong, E-mail: yinyulong@isa.ac.cn [State Key Laboratory of Food Science and Technology and School of Food Science, Nanchang University, Nanchang 330047 (China); Institute of Subtropical Agriculture, Chinese Academy of Sciences, Changsha 410125 (China)

    2016-01-22

    Acute or chronic hepatic injury is a common pathology worldwide. Mitochondrial dysfunction and the depletion of adenosine triphosphate (ATP) play important roles in liver injury. Chlorogenic acids (CGA) are some of the most abundant phenolic acids in human diet. This study was designed to test the hypothesis that CGA may protect against chronic lipopolysaccharide (LPS)-induced liver injury by modulating mitochondrial energy generation. CGA decreased the activities of serum alanine aminotransferase, aspartate aminotransferase and alkaline phosphatase. The contents of ATP and adenosine monophosphate (AMP), as well as the ratio of AMP/ATP, were increased after CGA supplementation. The activities of enzymes that are involved in glycolysis were reduced, while those of enzymes involved in oxidative phosphorylation were increased. Moreover, phosphorylated AMP-activated protein kinase (AMPK), and mRNA levels of AMPK-α, peroxisome proliferator-activated receptor-gamma coactivator 1α (PGC-1α), nuclear respiratory factor 1, and mitochondrial DNA transcription factor A were increased after CGA supplementation. Collectively, these findings suggest that the hepatoprotective effect of CGA might be associated with enhanced ATP production, the stimulation of mitochondrial oxidative phosphorylation and the inhibition of glycolysis. - Highlights: • Dietary supplementation with chlorogenic acid (CGA) improved endotoxin-induced liver injury. • Chlorogenic acid enhances ATP increase and shifts energy metabolism, which is correlated with up-regulation AMPK and PGC-1α. • The possible mechanism of CGA on mitochondrial biogenesis was correlated with up-regulation AMPK and PGC-1α.

  6. In vivo imaging of the effect of LPS on arterial endothelial cells: molecular imaging of heat shock protein 60 expression

    NARCIS (Netherlands)

    Wick, M.C.; Mayerl, Christina; Backovic, Aleksandar; Zee, R. van der; Jaschke, Werner; Dietrich, Hermann; Wick, G.

    2008-01-01

    Bacterial endotoxins are known as stress factors for endothelial cells. In 20 normocholesterolemic New Zealand White (NZW) rabbits, endothelial stress was induced by intravenous (i.v.) injection of lipopolysaccharide (LPS), while eight NZW rabbits were sham-treated or served as untreated

  7. LPS Catch and Effort Estimation

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Data collected from the LPS dockside (LPIS) and the LPS telephone (LPTS) surveys are combined to produce estimates of total recreational catch, landings, and fishing...

  8. Detection of Endotoxin Contamination of Graphene Based Materials Using the TNF-α Expression Test and Guidelines for Endotoxin-Free Graphene Oxide Production.

    Directory of Open Access Journals (Sweden)

    Sourav P Mukherjee

    Full Text Available Nanomaterials may be contaminated with bacterial endotoxin during production and handling, which may confound toxicological testing of these materials, not least when assessing for immunotoxicity. In the present study, we evaluated the conventional Limulus amebocyte lysate (LAL assay for endotoxin detection in graphene based material (GBM samples, including graphene oxide (GO and few-layered graphene (FLG. Our results showed that some GO samples interfered with various formats of the LAL assay. To overcome this problem, we developed a TNF-α expression test (TET using primary human monocyte-derived macrophages incubated in the presence or absence of the endotoxin inhibitor, polymyxin B sulfate, and found that this assay, performed with non-cytotoxic doses of the GBM samples, enabled unequivocal detection of endotoxin with a sensitivity that is comparable to the LAL assay. FLG also triggered TNF-α production in the presence of the LPS inhibitor, pointing to an intrinsic pro-inflammatory effect. Finally, we present guidelines for the preparation of endotoxin-free GO, validated by using the TET.

  9. Cardiac Function and Circulating Cytokines Following Endotoxin Exposure in Neonatal Mice

    Science.gov (United States)

    Mukherjee, Rupak; McQuinn, Tim C.; Dugan, Melissa A.; Saul, J. Philip; Spinale, Francis G.

    2011-01-01

    Complications following cardiac surgery in neonates can occur due to activation of the inflammatory system. This study used lipopolysaccharide (LPS) endotoxin exposure to cause cytokine activation in neonatal mice and examine left ventricular (LV) function as well as the effects of antioxidant treatment on cytokine levels. Neonatal mice (6 day old) were injected with either 25 mg/kg LPS (n=13) or phosphate buffered saline (PBS, n=14) and LV function (echocardiography) was measured at 4 hours. Plasma levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-4, IL-6, and IL-10 were measured at 30 min, 1, 2, and 4 hours after injection (n=5 mice per group). Effects of pretreatment with N-acetylcysteine (NAC, 50 mg/kg) on cytokine levels were examined at 2 and 4 hours following PBS or LPS (n=5 mice per group). Four hours after LPS heart rate was increased (434±14 vs. 405±14 bpm, p<0.05). LV end-diastolic dimension and ejection time were reduced with LPS (both p<0.05). LPS exposure increased plasma TNF-α, IL-6, and IL-10 levels. NAC pretreatment attenuated the increases in TNF-α and IL-6 levels, but augmented IL-10 levels at 2 hours post-LPS. LPS exposure altered cardiac performance and activated cytokines in neonatal mice, which may be ameliorated using antioxidants. PMID:20613681

  10. MiR-146b Mediates Endotoxin Tolerance in Human Phagocytes

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    Tiziana Ada Renzi

    2015-01-01

    Full Text Available A proper regulation of the innate immune response is fundamental to keep the immune system in check and avoid a chronic status of inflammation. As they act as negative modulators of TLR signaling pathways, miRNAs have been recently involved in the control of the inflammatory response. However, their role in the context of endotoxin tolerance is just beginning to be explored. We here show that miR-146b is upregulated in human monocytes tolerized by LPS, IL-10, or TGFβ priming and demonstrate that its transcription is driven by STAT3 and RUNX3, key factors downstream of IL-10 and TGFβ signaling. Our study also found that IFNγ, known to revert LPS tolerant state, inhibits miR-146b expression. Finally, we provide evidence that miR-146b levels have a profound effect on the tolerant state, thus candidating miR-146b as a molecular mediator of endotoxin tolerance.

  11. Prenatal endotoxin exposure alters behavioural pain responses to lipopolysaccharide in adult offspring.

    Science.gov (United States)

    Hodyl, Nicolette A; Walker, F Rohan; Krivanek, Klara M; Clifton, Vicki L; Hodgson, Deborah M

    2010-05-11

    Evidence suggests that exposure to bacterial endotoxin in early life can alter the production of pro-inflammatory cytokines in later life. This phenomenon may have significant consequences for pain and pain related behaviours as pro-inflammatory cytokines heighten pain sensitivity. This association has yet to be examined. As such, the aim of the present study was to characterize pain behaviours in adult rat offspring following prenatal endotoxin (PE) exposure. Pregnant F344 rats received endotoxin (200microg/kg, s.c.) or saline on gestational days 16, 18 and 20. Pain thresholds were assessed in the adult PE offspring (n=23) and control offspring (n=24) prior to and 4h following administration of lipopolysaccharide (LPS; 100microg/kg, s.c.). Three assays of pain were employed - the hot plate, tail immersion and von Frey tests. Results demonstrated sex-specific effects of prenatal endotoxin on the offspring, with PE males displaying unaltered pain thresholds on the von Frey test post-LPS administration (p<0.01), while male control offspring (n=24) displayed the expected hyperalgesia. Male PE offspring also displayed increased pain thresholds on the tail immersion test (p<0.01), while no change in pain sensitivity was observed in control males following LPS exposure. No difference in response was observed between the female PE and control offspring on the von Frey test, however PE female offspring displayed increased thresholds on the tail immersion test compared to baseline - an effect not observed in the control female offspring. Pain sensitivity on the hot plate test was unaffected by prenatal exposure to endotoxin. These data suggest that prenatal exposure to products associated with bacterial infection have the capacity to alter pain responses, which are evident in the adult offspring. Copyright 2010 Elsevier Inc. All rights reserved.

  12. Glucocorticoid Antagonism by Endotoxin: Biological Effects during Stress and Basis for Inhibition of Phosphoenolpyruvate Carboxykinase

    Science.gov (United States)

    1985-08-01

    monkeys (83), but not in rats (81), piglets (14S) or Yucatan minipigs (57). The lLhal effects of LPS and RD-LPS were also studied during stress conditions...gluconeogenesis and the hypoglycemia of endotoxin shock. Am. J. Physiol. 227: 778-781. 5S. Foster , 0.0., P.D. Ray and H.A. Lardy. 1366. Studies on the...Lardy, R.C. Nordlie and 0.0. Foster . ISS3. Metabolic and hormonal control of phosphoenolpyruvate carboxykinase and malic enzyme in rat liver. J. Biol

  13. Changes in endotoxin levels in T2DM subjects on anti-diabetic therapies

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    Kumar Sudhesh

    2009-04-01

    Full Text Available Abstract Introduction Chronic low-grade inflammation is a significant factor in the development of obesity associated diabetes. This is supported by recent studies suggesting endotoxin, derived from gut flora, may be key to the development of inflammation by stimulating the secretion of an adverse cytokine profile from adipose tissue. Aims The study investigated the relationship between endotoxin and various metabolic parameters of diabetic patients to determine if anti-diabetic therapies exerted a significant effect on endotoxin levels and adipocytokine profiles. Methods Fasting blood samples were collected from consenting Saudi Arabian patients (BMI: 30.2 ± (SD5.6 kg/m2, n = 413, consisting of non-diabetics (ND: n = 67 and T2DM subjects (n = 346. The diabetics were divided into 5 subgroups based on their 1 year treatment regimes: diet-controlled (n = 36, metformin (n = 141, rosiglitazone (RSG: n = 22, a combined fixed dose of metformin/rosiglitazone (met/RSG n = 100 and insulin (n = 47. Lipid profiles, fasting plasma glucose, insulin, adiponectin, resistin, TNF-α, leptin, C-reactive protein (CRP and endotoxin concentrations were determined. Results Regression analyses revealed significant correlations between endotoxin levels and triglycerides (R2 = 0.42; p 2 = 0.10; p 2 = 0.076; p 2 = 0.032; p 2 = 0.055; p Conclusion We conclude that sub-clinical inflammation in T2DM may, in part, be mediated by circulating endotoxin. Furthermore, that whilst the endotoxin and adipocytokine profiles of diabetic patients treated with different therapies were comparable, the RSG group demonstrated significant differences in both adiponectin and endotoxin levels. We confirm an association between endotoxin and serum insulin and triglycerides and an inverse relationship with HDL. Lower endotoxin and higher adiponectin in the groups treated with RSG may be related and indicate another mechanism for the effect of RSG on insulin sensitivity.

  14. LPS-induced delayed preconditioning is mediated by Hsp90 and involves the heat shock response in mouse kidney.

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    Tamás Kaucsár

    Full Text Available We and others demonstrated previously that preconditioning with endotoxin (LPS protected from a subsequent lethal LPS challenge or from renal ischemia-reperfusion injury (IRI. LPS is effective in evoking the heat shock response, an ancient and essential cellular defense mechanism, which plays a role in resistance to, and recovery from diseases. Here, by using the pharmacological Hsp90 inhibitor novobiocin (NB, we investigated the role of Hsp90 and the heat shock response in LPS-induced delayed renal preconditioning.Male C57BL/6 mice were treated with preconditioning (P: 2 mg/kg, i.p. and subsequent lethal (L: 10 mg/kg, i.p. doses of LPS alone or in combination with NB (100 mg/kg, i.p.. Controls received saline (C or NB.Preconditioning LPS conferred protection from a subsequent lethal LPS treatment. Importantly, the protective effect of LPS preconditioning was completely abolished by a concomitant treatment with NB. LPS induced a marked heat shock protein increase as demonstrated by Western blots of Hsp70 and Hsp90. NB alone also stimulated Hsp70 and Hsp90 mRNA but not protein expression. However, Hsp70 and Hsp90 protein induction in LPS-treated mice was abolished by a concomitant NB treatment, demonstrating a NB-induced impairment of the heat shock response to LPS preconditioning.LPS-induced heat shock protein induction and tolerance to a subsequent lethal LPS treatment was prevented by the Hsp90 inhibitor, novobiocin. Our findings demonstrate a critical role of Hsp90 in LPS signaling, and a potential involvement of the heat shock response in LPS-induced preconditioning.

  15. Low level bacterial endotoxin activates two distinct signaling pathways in human peripheral blood mononuclear cells

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    Weintraub Neal L

    2011-02-01

    Full Text Available Abstract Background Bacterial endotoxin, long recognized as a potent pro-inflammatory mediator in acute infectious processes, has more recently been identified as a risk factor for atherosclerosis and other cardiovascular diseases. When endotoxin enters the bloodstream, one of the first cells activated is the circulating monocyte, which exhibits a wide range of pro-inflammatory responses. Methods We studied the effect of low doses of E. coli LPS on IL-8 release and superoxide formation by freshly isolated human peripheral blood mononuclear cells (PBMC. Results IL-8 release was consistently detectable at 10 pg/ml of endotoxin, reaching a maximum at 1 ng/ml, and was exclusively produced by monocytes; the lymphocytes neither produced IL-8, nor affected monocyte IL-8 release. Superoxide production was detectable at 30 pg/ml of endotoxin, reaching a maximum at 3 ng/ml. Peak respiratory burst activity was seen at 15-20 min, and superoxide levels returned to baseline by 1 h. IL-8 release was dependent on both membrane-associated CD14 (mCD14 and Toll-like receptor 4 (TLR4. Superoxide production was dependent on the presence of LBP, but was not significantly affected by a blocking antibody to TLR4. Moreover, treatment with lovastatin inhibited LPS-dependent IL-8 release and superoxide production. Conclusions These findings suggest that IL-8 release and the respiratory burst are regulated by distinct endotoxin-dependent signaling pathways in PBMC in low level of endotoxin exposure. Selectively modulating these pathways could lead to new approaches to treat chronic inflammatory diseases, such as atherosclerosis, while preserving the capacity of monocytes to respond to acute bacterial infections.

  16. Factors affecting endotoxin release from the cell surface of avian strains of Pasteurella multocida.

    Science.gov (United States)

    Lee, M D; Glisson, J R; Wooley, R E

    1992-06-15

    Two avian strains of Pasteurella multocida, a vaccine strain and a virulent field isolate, were investigated to determine their propensity to release endotoxin from the cell surface. Both organisms released comparable amounts of endotoxin when plasma complement proteins were present, however the virulent strain did so without the loss of viability that occurred in the vaccine strain. Blocking complement activity decreased the ability of plasma to elicit endotoxin release from the bacteria. When the cells were treated with divalent metal chelators such as trans-1, 2-diaminocyclohexane-N,N,N1,N1-tetraacetic acid (CDTA), more endotoxin was released from the vaccine strain than from the virulent isolate. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of purified lipopolysaccharide (LPS) from both strains revealed virtually identical patterns. Both had patterns considered typical of rough LPS. Challenge studies in 8 weeks old turkeys showed that the field strain induced endotoxemia of longer duration than the vaccine strain and produced greater mortality.

  17. Allergen endotoxins induce T-cell-dependent and non-IgE-mediated nasal hypersensitivity in mice.

    Science.gov (United States)

    Iwasaki, Naruhito; Matsushita, Kazufumi; Fukuoka, Ayumi; Nakahira, Masakiyo; Matsumoto, Makoto; Akasaki, Shoko; Yasuda, Koubun; Shimizu, Takeshi; Yoshimoto, Tomohiro

    2017-01-01

    Allergen-mediated cross-linking of IgE on mast cells/basophils is a well-recognized trigger for type 1 allergic diseases such as allergic rhinitis (AR). However, allergens may not be the sole trigger for AR, and several allergic-like reactions are induced by non-IgE-mediated mechanisms. We sought to describe a novel non-IgE-mediated, endotoxin-triggered nasal type-1-hypersensitivity-like reaction in mice. To investigate whether endotoxin affects sneezing responses, mice were intraperitoneally immunized with ovalbumin (OVA), then nasally challenged with endotoxin-free or endotoxin-containing OVA. To investigate the role of T cells and mechanisms of the endotoxin-induced response, mice were adoptively transferred with in vitro-differentiated OVA-specific TH2 cells, then nasally challenged with endotoxin-free or endotoxin-containing OVA. Endotoxin-containing, but not endotoxin-free, OVA elicited sneezing responses in mice independent from IgE-mediated signaling. OVA-specific TH2 cell adoptive transfer to mice demonstrated that local activation of antigen-specific TH2 cells was required for the response. The Toll-like receptor 4-myeloid differentiation factor 88 signaling pathway was indispensable for endotoxin-containing OVA-elicited rhinitis. In addition, LPS directly triggered sneezing responses in OVA-specific TH2-transferred and nasally endotoxin-free OVA-primed mice. Although antihistamines suppressed sneezing responses, mast-cell/basophil-depleted mice had normal sneezing responses to endotoxin-containing OVA. Clodronate treatment abrogated endotoxin-containing OVA-elicited rhinitis, suggesting the involvement of monocytes/macrophages in this response. Antigen-specific nasal activation of CD4+ T cells followed by endotoxin exposure induces mast cell/basophil-independent histamine release in the nose that elicits sneezing responses. Thus, environmental or nasal residential bacteria may exacerbate AR symptoms. In addition, this novel phenomenon might explain

  18. Endotoxin testing of a wound debridement device containing medicinal Lucilia sericata larvae.

    Science.gov (United States)

    Pickles, Samantha F; Pritchard, David I

    2017-05-01

    Alimentary products of medicinal Lucilia sericata larvae are studied to determine their mechanisms of action, particularly in the contexts of wound debridement and disinfection. Furthermore, the larvae can be applied to patients in contained medical devices (such as the BioBag; BioMonde). Here, we tested the materials and larval content of the most commonly used debridement device (the "BB-50") to explore the possibility that endotoxins may be contributing to the bio-activity of the product, given that endotoxins are potent stimulants of cellular activation. Using standardised protocols to collect larval alimentary products (LAP), we proceeded to determine residual endotoxin levels in LAP derived from the device, before and after the neutralisation of interfering enzymatic activity. The debridement device and its associated larval content was not a significant source of lipopolysaccharide (LPS) activity. However, it is clear from these experiments that a failure to remove the confounding serine proteinase activity would have resulted in spuriously high and erroneous results. The residual LPS levels detected are unlikely to be active in wound healing assays, following cross-referencing to publications where LPS at much higher levels has been shown to have positive and negative effects on processes associated with wound repair and tissue regeneration. © 2017 by the Wound Healing Society.

  19. Comparison of bacterial endotoxin lipopolysaccharide concentrations in the blood, ovarian follicular fluid and uterine fluid: a clinical case of bovine metritis

    Science.gov (United States)

    MAGATA, Fumie; ISHIDA, Yoshikazu; MIYAMOTO, Akio; FURUOKA, Hidefumi; INOKUMA, Hisashi; SHIMIZU, Takashi

    2014-01-01

    We investigated the concentration of the bacterial endotoxin lipopolysaccharide (LPS) in the blood, ovarian follicular fluid and uterine fluid of a clinical case of bovine metritis. A 2-year-old lactating Holstein cow exhibited continuous fever >39.5°C for more than 2 weeks after normal calving. The cow produced a fetid, watery, red-brown uterine discharge from the vagina and was diagnosed with metritis. The LPS concentrations in plasma and uterine fluid were 0.94 and 6.34 endotoxin units (EU)/ml, respectively. One of seven follicles showed an extremely high level of LPS (12.40 EU/ml) compared to the other follicles (0.62–0.97 EU/ml). These results might suggest the presence of high concentration of LPS in follicles in cows with postpartum metritis. PMID:25223344

  20. Pyometra in Bitches Induces Elevated Plasma Endotoxin and Prostaglandin F2α Metabolite Levels

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    Hagman R

    2006-03-01

    Full Text Available Endotoxemia in bitches with pyometra can cause severe systemic effects directly or via the release of inflammatory mediators. Plasma endotoxin concentrations were measured in ten bitches suffering from pyometra with moderately to severely deteriorated general condition, and in nine bitches admitted to surgery for non-infectious reasons. Endotoxin samples were taken on five occasions before, during and after surgery. In addition, urine and uterine bacteriology was performed and hematological, blood biochemical parameters, prostaglandin F2α metabolite 15-ketodihydro-PGF2α (PG-metabolite, progesterone and oestradiol (E2-17β levels were analysed. The results confirm significantly increased plasma levels of endotoxin in bitches with pyometra and support previous reports of endotoxin involvement in the pathogenesis of the disease. Plasma concentrations of PG-metabolite were elevated in pyometra bitches and provide a good indicator of endotoxin release since the concentrations were significantly correlated to the endotoxin levels and many other hematological and chemistry parameters. The γ-globulin serum protein electrophoresis fraction and analysis of PG-metabolite can be valuable in the diagnosis of endotoxin involvement if a reliable, rapid and cost-effective test for PG-metabolite analysis becomes readily available in the future. Treatment inhibiting prostaglandin biosynthesis and related compounds could be beneficial for bitches suffering from pyometra.

  1. Role of pulmonary intravascular macrophages in endotoxin-induced lung inflammation and mortality in a rat model

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    Duke Tanya

    2008-10-01

    Full Text Available Abstract Background Bile-duct ligated (BDL rats recruit pulmonary intravascular macrophages (PIMs and are highly susceptible to endotoxin-induced mortality. The mechanisms of this enhanced susceptibility and mortality in BDL rats, which are used as a model of hepato-pulmonary syndrome, remain unknown. We tested a hypothesis that recruited PIMs promote endotoxin-induced mortality in a rat model. Methods Rats were subjected to BDL to induce PIM recruitment followed by treatment with gadolinium chloride (GC to deplete PIMs. Normal and BDL rats were treated intravenously with E. coli lipopolysaccharide (LPS with or without GC pre-treatment followed by collection and analyses of lungs for histopathology, electron microscopy and cytokine quantification. Results BDL rats recruited PIMs without any change in the expression of IL-1β, TNF-α and IL-10. GC caused reduction in PIMs at 48 hours post-treatment (P E. coli LPS died within 3 hours of the challenge while the normal LPS-treated rats were euthanized at 6 hours after the LPS treatment. GC treatment of rats 6 hours or 48 hours before LPS challenge resulted in 80% (1/5 and 100% (0/5 survival, respectively, at 6 hours post-LPS treatment. Lungs from BDL+LPS rats showed large areas of perivascular hemorrhages compared to those pre-treated with GC. Concentrations of IL-1β, TNF-α and IL-10 were increased in lungs of BDL+LPS rats compared to BDL rats treated with GC 48 hours but not 6 hours before LPS (P Conclusion We conclude that PIMs increase susceptibility for LPS-induced lung injury and mortality in this model, which is blocked by a reduction in their numbers or their inactivation.

  2. Endotoxin Studies And Biosolids Stabilization Research

    Science.gov (United States)

    This presentation has three parts; a review of bench-scale endotoxin research, a review of observations from a field scale endotoxin release study, and discussion of biosolids stabilization and characterization by PLFA/FAME microbial community analysis. Endotoxins are part of th...

  3. Extraction, separation and characterization of endotoxins in water samples using solid phase extraction and capillary electrophoresis-laser induced fluorescence.

    Science.gov (United States)

    Fung, Fun Man; Su, Min; Feng, Hua-Tao; Li, Sam Fong Yau

    2017-09-07

    This study focuses on one of the key environmental threats, endotoxins, also known as lipopolysaccharides (LPS). A capillary electrophoresis method in combination with laser induced fluorescence (LIF) detection was developed for the analysis of endotoxins from 16 different bacterial strains. LPSs were derivatized with the amino-reactive fluorescent dye, fluorescein isothiocyanate (FITC), separated by capillary zone electrophoresis (CZE) under the optimized conditions with the use of 50 mM sodium tetraborate buffer (pH 9.30), and detected by LIF detector. To improve the sensitivity of CZE-LIF detection for the determination of trace amounts of endotoxins and to remove possible interference materials in environmental samples, a solid phase extraction (SPE) pre-concentration technique was applied successfully. The SPE targeted at polysaccharide moieties of LPSs and showed LPS enrichment effects too. CE migration time could also reveal the O-antigen chain lengths of LPSs. This CE method and SPE pretreatment showed linearity at 99.84%, and repeatabilities at 8.44% and 11.0% for endotoxins from E. Coli O55:B5 and E. Coli O26:B6. The limit of detection (LOD) could reach around 5 ng/mL at optimized condition. The method was applied successfully to the determination of LPS levels in tap water and wastewater, and demonstrated sensitive, reproducible and reliable results.

  4. LPS-induced galectin-3 oligomerization results in enhancement of neutrophil activation.

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    Marise Lopes Fermino

    Full Text Available Galectin-3 (Gal 3 is a glycan-binding protein that can be secreted by activated macrophages and mast cells at inflammation sites and plays an important role in inflammatory diseases caused by Bacteria and their products, such as lipopolysaccharides (LPS. Although it is well established that Gal 3 can interact with LPS, the pathophysiological importance of LPS/Gal 3 interactions is not fully understood. Data presented herein demonstrate for the first time that the interaction of Gal 3, either via its carbohydrate binding C-terminal domain or via its N-terminal part, with LPS from different bacterial strains, enhances the LPS-mediated neutrophil activation in vitro. Gal 3 allowed low LPS concentrations (1 µg/mL without serum, 1 ng/mL with serum to upregulate CD11b expression and reactive oxygen species (ROS generation on human neutrophils in vitro and drastically enhanced the binding efficiency of LPS to the neutrophil surface. These effects required LPS preincubation with Gal 3, before neutrophil stimulation and involved specific Gal 3/LPS interaction. A C-terminal Gal-3 fragment, which retains the lectin domain but lacks the N-terminal part, was still able to bind both to Escherichia coli LPS and to neutrophils, but had lost the ability to enhance neutrophil response to LPS. This result emphasizes the importance of an N-terminus-mediated Gal 3 oligomerization induced by its interaction with LPS. Finally we demonstrated that Balb/C mice were more susceptible to LPS-mediated shock when LPS was pretreated with Gal 3. Altogether, these results suggest that multimeric interactions between Gal 3 oligomers and LPS potentiate its pro-inflammatory effects on neutrophils.

  5. Sialylation of Porphyromonas gingivalis LPS and its effect on bacterial-host interactions.

    Science.gov (United States)

    Zaric, Svetislav S; Lappin, Mark J; Fulton, Catherine R; Lundy, Fionnuala T; Coulter, Wilson A; Irwin, Christopher R

    2017-04-01

    Porphyromonas gingivalis produces different LPS isoforms with significant structural variations of their lipid A and O-antigen moieties that can affect its pro-inflammatory and bone-resorbing potential. We show here, for the first time, that P. gingivalis LPS isolated from W83 strain is highly sialylated and possesses significantly reduced inflammatory potential compared with less sialylated ATCC 33277 strain LPS. Nevertheless, the reduction in the endotoxin activity is not mediated by the presence of sialic acid LPS moieties as the sialic acid-free LPS produced by the mutant W83 strain exhibits a similar inflammatory potential to the wild type strain. Furthermore, our findings suggest that the interaction between the sialic acid LPS moieties and the inhibitory CD33 receptor is prevented by endogenously expressed sialic acid on the surface of THP-1 cells that cannot be out-competed by sialic acid containing P. gingivalis LPS. The present study also highlights the importance of endogenous sialic acid as a 'self-associated molecular pattern' and CD33 receptors in modulation of innate immune response as human gingival fibroblasts, which do not express CD33 receptors, and desialylated THP-1 cells have both been found to have much higher spontaneous IL-8 production than naïve THP-1 cells.

  6. Effect of endotoxin on lipid peroxidation in vivo in selenium and vitamin E deficient rats

    Energy Technology Data Exchange (ETDEWEB)

    Sword, J.T.; Pope, A.L.; Hoekstra, W.G.

    1986-03-01

    The authors have used respiratory ethane production by selenium (Se) and vitamin E (E) deficient rats, an index of lipid peroxidation, to identify oxidant stressors which might precipitate sudden tissue degeneration in deficient animals. Other studies have suggested that endotoxin (gram-negative bacterial lipopolysaccharide-LPS) might be such an oxidant stressor, especially in the lungs. Male weanling rats were fed a Se and E deficient diet for about 80 days. Rats were injected ip with Salmonella typhimurium LPS (.25, .5, or 1.0 mg/kg) or saline, and respiratory ethane was collected for 16 hr. In a representative experiment, mean rate of ethane production (nm/100g/hr) was increased (p < .01) by LPS: saline, .48 +/- .04 (SEM); .25 mg LPS/kg, 1.30 +/- .17; .5, 1.47 +/- .18 and 1.0, 1.68 +/- .18. E. coli and S. minnesota LPS gave similar results. Rats fed a supplemented diet (.2 ppm Se and 200 IU E/kg diet) produced less (p < .01) ethane: saline, .068 +/- .009 and .5 mg LPS/kg, .114 +/- .01. Over all experiments LPS produced a small yet significant increase in ethane in rats receiving Se or E supplementation but produced a marked increase in unsupplemented rats. In further studies with LPS treated rats, Se supplementation alone was 73%, and E supplementation alone 99% as effective as Se + E. These results showed that LPS stimulates lipid peroxidation in Se and E deficient rats and that infections may initiate oxidative cell damage in deficient animals. E was more protective than Se against LPS-induced peroxidation.

  7. Anti-endotoxin and anti-inflammatory effects of Chinese herbal medicinal alkaloid ingredients in vivo.

    Science.gov (United States)

    Hu, Yiyi; Mao, Aihua; Yu, Zhengyu; He, Kongwang

    2016-10-01

    The aim of the research was to investigate the anti-endotoxin and anti-inflammatory effects of sinomenine, fangchinoline, stachydrine, chuanxionggzine, oxymartrine, and evodiamine alkaloids commonly found in Chinese herbal medicines. In an endotoxin (LPS) control group, each mouse was challenged with 1 mg LPS/kg by intraperitoneal (IP) injection. In high-, middle- and low-dose alkaloid groups, mice were initially challenged with 1 mg LPS/kg by IP injection and, 3 h later, dosed intramuscularly (IM) with one of the six alkaloids at one of three levels (1, 5, or 10 mg/kg body weight). In the drug control group, mice were dosed IM with 10 mg/kg body weight of a given alkaloid; mice in a naïve control group were administered the same volume of normal saline. The results revealed the six alkaloids could reduce the incidence/severity of LPS- induced toxicities, e.g., body temperature elevation, weight loss, systemic inflammation, multiple organ dysfunction. Taken together, the data suggested to us that these alkaloids might effectively regulate inflammatory responses and have a potential to be used in anti-endotoxin therapies. Copyright © 2016 Elsevier Ltd. All rights reserved.

  8. Endotoxin emissions from commercial composting activities

    Science.gov (United States)

    2009-01-01

    This paper describes an exploratory study of endotoxin emissions and dispersal from a commercial composting facility. Replicated samples of air were taken by filtration at different locations around the facility on 10 occasions. Measurements were made of endotoxin and associated culturable microorganisms. The inflammatory response of cell cultures exposed to extracts from the filters was measured. Endotoxin was detected in elevated concentrations close to composting activities. A secondary peak, of lesser magnitude than the peak at source was detected at 100-150 m downwind of the site boundary. Unexpectedly high concentrations of endotoxin were measured at the most distant downwind sampling point. Extracted endotoxin was found to stimulate human monocytes and a human lung epithelial cell line to produce significant amounts of pro-inflammatory cytokines. On a weight basis, endotoxin extracted from the composting source has a greater inflammatory cytokine inducing effect than commercial E. coli endotoxin. PMID:20102594

  9. Long-lasting pro-inflammatory suppression of microglia by LPS-preconditioning is mediated by RelB-dependent epigenetic silencing

    NARCIS (Netherlands)

    Schaafsma, W.; Zhang, X.; van Zomeren, K. C.; Jacobs, S.; Georgieva, P. B.; Wolf, S. A.; Kettenmann, H.; Janova, H.; Saiepour, N.; Hanisch, U. -K.; Meerlo, P.; van den Elsen, P. J.; Brouwer, N.; Boddeke, H. W. G. M.; Eggen, B. J. L.

    Microglia, the innate immune cells of the central nervous system (CNS), react to endotoxins like bacterial lipopolysaccharides (LPS) with a pronounced inflammatory response. To avoid excess damage to the CNS, the microglia inflammatory response needs to be tightly regulated. Here we report that a

  10. Long-lasting pro-inflammatory suppression of microglia by LPS-preconditioning is mediated by RelB-dependent epigenetic silencing

    NARCIS (Netherlands)

    Schaafsma, W.; Zhang, X.; van Zomeren, K.C.; Jacobs, S.; Georgieva, P.B.; Wolf, S.A.; Kettenmann, H.; Janova, H.; Saiepour, N.; Hanisch, U.K.; Meerlo, P.; van den Elsen, P.J.; Brouwer, N.; Boddeke, H.W.G.M.; Eggen, B.J.L.

    2015-01-01

    Microglia, the innate immune cells of the central nervous system (CNS), react to endotoxins like bacterial lipopolysaccharides (LPS) with a pronounced inflammatory response. To avoid excess damage to the CNS, the microglia inflammatory response needs to be tightly regulated. Here we report that a

  11. Inhibiting TNF-α signaling does not attenuate induction of endotoxin tolerance

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    Loosbroock C

    2014-12-01

    Full Text Available Christopher Loosbroock, Kenneth W Hunter Department of Microbiology and Immunology, University of Nevada School of Medicine, Reno, NV, USA Abstract: Tumor necrosis factor-alpha (TNF-α is a central mediator of inflammatory responses elicited by Toll-like receptor agonists, such as the Gram-negative bacterial outer membrane antigen lipopolysaccharide (LPS. TNF-α is responsible for altering vascular permeability and activating infiltrating inflammatory cells, such as monocytes and neutrophils. Interestingly, TNF-α has also demonstrated the ability to induce tolerance to subsequent challenges with TNF-α or LPS in monocyte and macrophage cell populations. Tolerance is characterized by the inability to mount a typical inflammatory response during subsequent challenges following the initial exposure to an inflammatory mediator such as LPS. The ability of TNF-α to induce a tolerant-like state with regard to LPS is most likely a regulatory mechanism to prevent excessive inflammation. We hypothesized that the induction of tolerance or the degree of tolerance is dependent upon the production of TNF-α during the primary response to LPS. To investigate TNF-α-dependent tolerance, human monocytic THP-1 cells were treated with TNF-α-neutralizing antibodies or antagonistic TNF-α receptor antibodies before primary LPS stimulation and then monitored for the production of TNF-α during the primary and challenge stimulation. During the primary stimulation, anti-TNF-α treatment effectively attenuated the production of TNF-α and interleukin-1β; however, this reduced production did not impact the induction of endotoxin tolerance. These results demonstrate that interfering with TNF-α signaling attenuates production of inflammatory cytokines without affecting the induction of tolerance. Keywords: endotoxin tolerance, lipopolysaccharide, tumor necrosis factor-alpha, anti-tumor necrosis factor-alpha, THP-1 cells

  12. Niacinamide abrogates the organ dysfunction and acute lung injury caused by endotoxin.

    Science.gov (United States)

    Kao, Shang-Jyh; Liu, Demeral David; Su, Chain-Fa; Chen, Hsing I

    2007-09-01

    Poly (ADP-ribose) synthabse (PARS) or polymerase (PARP) is a cytotoxic enzyme causing cellular damage. Niacinamide inhibits PARS or PARP. The present experiment tests the effects of niacinamide (NCA) on organ dysfunction and acute lung injury (ALI) following lipopolysaccharide (LPS). LPS was administered to anesthetized rats and to isolated rat lungs. In anesthetized rats, LPS caused systemic hypotension and increased biochemical factors, nitrate/nitrite (NOx), methyl guanidine (MG), tumor necrosis factoralpha (TNFalpha), and interleukin-1beta (IL-1beta). In isolated lungs, LPS increased lung weight (LW) to body weight ratio, LW gain, protein and dye tracer leakage, and capillary permeability. The insult also increased NOx, MG, TNFalpha, and IL-1beta in lung perfusate, while decreased adenosine triphosphate (ATP) content with an increase in PARP activity in lung tissue. Pathological examination revealed pulmonary edema with inflammatory cell infiltration. These changes were abrogated by posttreatment (30 min after LPS) with NCA. Following LPS, the inducible NO synthase (iNOS) mRNA expression was increased. NCA reduced the iNOS expression. Niacinamide exerts protective effects on the organ dysfunction and ALI caused by endotoxin. The mechanisms may be mediated through the inhibition on the PARP activity, iNOS expression and the subsequent suppression of NO, free radicals, and proinflammatory cytokines with restoration of ATP.

  13. Associations between endotoxin-induced metabolic changes and temperament in Brahman bulls.

    Science.gov (United States)

    Burdick Sanchez, N C; Carroll, J A; Randel, R D; Vann, R C; Welsh, T H

    2014-02-01

    The influence of temperament on the alteration of metabolic parameters in response to a lipopolysaccharide(LPS) challenge was investigated. Brahman bulls were selected based on temperament score. Bulls (10 months; 211±5kg BW; n = 6, 8 and 7 for Calm, Intermediate and Temperamental groups, respectively) were fitted with indwelling jugular catheters to evaluate peripheral blood concentrations of glucose, blood urea nitrogen (BUN),non-esterified fatty acids (NEFA), insulin, epinephrine and cortisol before and after LPS administration (0.5 μg/kg BW LPS). Feed intake was also recorded. Intermediate bulls consumed more feed than the Temperamental bulls during the challenge (p = 0.046). Pre-LPS glucose (p = 0.401) and BUN (p = 0.222) did not differ among the temperament groups. However, pre-LPS insulin (p = 0.023) was lower, whereas pre-LPS NEFA (p metabolic responses of Brahman bulls following a provocative endotoxin challenge.Specifically, Temperamental bulls may preferentially utilize an alternate energy source (i.e. NEFA) to a greater degree than do bulls of Calm and Intermediate temperaments. The use of circulating NEFA from lipolysis may reduce the negative metabolic consequences of an immune response by allowing for a prompt answer to increasing energy demands required during immunological challenge, compared with the time required for glycogenolysis and gluconeogenesis.

  14. Capture of lipopolysaccharide (endotoxin by the blood clot: a comparative study.

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    Margaret T Armstrong

    Full Text Available In vertebrates and arthropods, blood clotting involves the establishment of a plug of aggregated thrombocytes (the cellular clot and an extracellular fibrillar clot formed by the polymerization of the structural protein of the clot, which is fibrin in mammals, plasma lipoprotein in crustaceans, and coagulin in the horseshoe crab, Limulus polyphemus. Both elements of the clot function to staunch bleeding. Additionally, the extracellular clot functions as an agent of the innate immune system by providing a passive anti-microbial barrier and microbial entrapment device, which functions directly at the site of wounds to the integument. Here we show that, in addition to these passive functions in immunity, the plasma lipoprotein clot of lobster, the coagulin clot of Limulus, and both the platelet thrombus and the fibrin clot of mammals (human, mouse operate to capture lipopolysaccharide (LPS, endotoxin. The lipid A core of LPS is the principal agent of gram-negative septicemia, which is responsible for more than 100,000 human deaths annually in the United States and is similarly toxic to arthropods. Quantification using the Limulus Amebocyte Lysate (LAL test shows that clots capture significant quantities of LPS and fluorescent-labeled LPS can be seen by microscopy to decorate the clot fibrils. Thrombi generated in the living mouse accumulate LPS in vivo. It is suggested that capture of LPS released from gram-negative bacteria entrapped by the blood clot operates to protect against the disease that might be caused by its systemic dispersal.

  15. Capture of lipopolysaccharide (endotoxin) by the blood clot: a comparative study.

    Science.gov (United States)

    Armstrong, Margaret T; Rickles, Frederick R; Armstrong, Peter B

    2013-01-01

    In vertebrates and arthropods, blood clotting involves the establishment of a plug of aggregated thrombocytes (the cellular clot) and an extracellular fibrillar clot formed by the polymerization of the structural protein of the clot, which is fibrin in mammals, plasma lipoprotein in crustaceans, and coagulin in the horseshoe crab, Limulus polyphemus. Both elements of the clot function to staunch bleeding. Additionally, the extracellular clot functions as an agent of the innate immune system by providing a passive anti-microbial barrier and microbial entrapment device, which functions directly at the site of wounds to the integument. Here we show that, in addition to these passive functions in immunity, the plasma lipoprotein clot of lobster, the coagulin clot of Limulus, and both the platelet thrombus and the fibrin clot of mammals (human, mouse) operate to capture lipopolysaccharide (LPS, endotoxin). The lipid A core of LPS is the principal agent of gram-negative septicemia, which is responsible for more than 100,000 human deaths annually in the United States and is similarly toxic to arthropods. Quantification using the Limulus Amebocyte Lysate (LAL) test shows that clots capture significant quantities of LPS and fluorescent-labeled LPS can be seen by microscopy to decorate the clot fibrils. Thrombi generated in the living mouse accumulate LPS in vivo. It is suggested that capture of LPS released from gram-negative bacteria entrapped by the blood clot operates to protect against the disease that might be caused by its systemic dispersal.

  16. Microencapsulated Bifidobacterium longum subsp. infantis ATCC 15697 favorably modulates gut microbiota and reduces circulating endotoxins in F344 rats.

    Science.gov (United States)

    Rodes, Laetitia; Saha, Shyamali; Tomaro-Duchesneau, Catherine; Prakash, Satya

    2014-01-01

    The gut microbiota is a bacterial bioreactor whose composition is an asset for human health. However, circulating gut microbiota derived endotoxins cause metabolic endotoxemia, promoting metabolic and liver diseases. This study investigates the potential of orally delivered microencapsulated Bifidobacterium infantis ATCC 15697 to modulate the gut microbiota and reduce endotoxemia in F344 rats. The rats were gavaged daily with saline or microencapsulated B. infantis ATCC 15697. Following 38 days of supplementation, the treated rats showed a significant (P endotoxins. Thus, microencapsulated B. infantis ATCC 15697 modulates the gut microbiota and reduces colonic and serum endotoxins. Future preclinical studies should investigate the potential of the novel probiotic formulation in metabolic and liver diseases.

  17. NAC attenuates LPS-induced toxicity in aspirin-sensitized mouse macrophages via suppression of oxidative stress and mitochondrial dysfunction.

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    Haider Raza

    Full Text Available Bacterial endotoxin lipopolysaccharide (LPS induces the production of inflammatory cytokines and reactive oxygen species (ROS under in vivo and in vitro conditions. Acetylsalicylic acid (ASA, aspirin is a commonly used anti-inflammatory drug. Our aim was to study the effects of N-acetyl cysteine (NAC, an antioxidant precursor of GSH synthesis, on aspirin-sensitized macrophages treated with LPS. We investigated the effects of LPS alone and in conjunction with a sub-toxic concentration of ASA, on metabolic and oxidative stress, apoptosis, and mitochondrial function using J774.2 mouse macrophage cell line. Protection from LPS-induced toxicity by NAC was also studied. LPS alone markedly induced ROS production and oxidative stress in macrophage cells. When ASA was added to LPS-treated macrophages, the increase in oxidative stress was significantly higher than that with LPS alone. Similarly, alteration in glutathione-dependent redox metabolism was also observed in macrophages after treatment with LPS and ASA. The combination of LPS and ASA selectively altered the CYP 3A4, CYP 2E1 and CYP 1A1 catalytic activities. Mitochondrial respiratory complexes and ATP production were also inhibited by LPS-ASA treatment. Furthermore a higher apoptotic cell death was also observed in LPS-ASA treated macrophages. NAC pre-treatment showed protection against oxidative stress induced apoptosis and mitochondrial dysfunction. These effects are presumed, at least in part, to be associated with alterations in NF-κB/Nrf-2 mediated cell signaling. These results suggest that macrophages are more sensitive to LPS when challenged with ASA and that NAC pre-treatment protects the macrophages from these deleterious effects.

  18. Immobilization of ɛ-polylysine onto the probe surface for molecular adsorption type endotoxin detection system

    Science.gov (United States)

    Ooe, Katsutoshi; Tsuji, Akihito; Nishishita, Naoki; Hirano, Yoshiaki

    2007-04-01

    Patients with renal failure become not able to expel the waste product, and they accumulate the toxic products for themselves. They therefore must use the hemodialysis to weed out the metabolic decomposition product. Hemodialysis for chronic renal failure is the most popular treatment method with artificial organs. However, hemodialysis patients must continue the treatment throughout their life, the results of long term extracorporeal dialysis, those patients develop the various complications and diseases, for example, dialysis amyloidosis etc. Dialysis amyloidosis is one of the refractory complications, and the prevention of this complication is important. Recently, endotoxin is thought to be the most likely cause of the complication. Endotoxin is one of the major cell wall components of gram-negative bacteria, and it forms the complex with proteins and lipopolysaccharide (LPS). It has various biological activities, e.g. attack of fever, when it gets mixed into human blood. In addition, it is known that large amount of endotoxin exists in living environment, and medicine is often contaminated with endotoxin. When contaminated dialyzing fluids are used to hemodialysis, above-mentioned dialysis amyloidosis is developed. Therefore, it is important that the detection and removal of endotoxin from dialyzing fluids. Until now, the measurement methods using Limulus Amebosyte Lysate (LAL) reagent were carried out as the tests for the presence of endotoxin. However, these methods include several different varieties of measurement techniques. The following are examples of them, gelatinization method, turbidimetric assay method, colorimetric assay method and fluoroscopic method. However, these techniques needed 30-60 minutes for the measurement. From these facts, they are not able to use as a "real-time endotoxin detector". The detection of endotoxin has needed to carry out immediately, for that reason, a new "real-time" detection method is desired. We focused attention to

  19. Endotoxin-induced reduction of social investigation by mice: interaction with amphetamine and anti-inflammatory drugs.

    Science.gov (United States)

    Fishkin, R J; Winslow, J T

    1997-08-01

    Previous studies indicate that some aspects of endotoxin-induced sickness behavior in rats may be mediated by interleukin-1 stimulated events and can be attenuated by corticosteroids, cyclooxygenase inhibitors and the interleukin-1-receptor antagonist. In the current studies, we replicate and extend these findings in adult male mice. A relatively low dose of lipopolysaccharide (LPS; 15 micrograms/kg, IP) was used to reliably induce a 50-60% reduction in the social investigation of a juvenile conspecific at 2-3 h after injection. Amphetamine (2.0-4.0 mg/kg, IP, 30 min pre-LPS) exacerbated LPS-induced decreases in investigation. Administration of methylprednisolone (10-30 mg/kg, IP), indomethacin (3-30 mg/kg, IP), and ibuprofen (1-100 mg/kg, IP) 1 h before LPS significantly reduced LPS-induced sickness behavior at several doses. Dexamethasone (0.1-10 mg/kg, IP) partially antagonized sickness. Representative flavonoids rohitukine (0.01-100.0 mg/kg, IP) and chrysin (0.01-10 mg/kg, IP) also antagonized LPS-induced deficits in social investigation. These studies replicate and extend previous studies in rat to demonstrate systematic effects of low doses of LPS, antagonism by anti-inflammatory drugs and enhancement of LPS effects by amphetamine. The latter findings are consistent with a modulatory role for adrenergic activation on interleukin-1 release stimulated by endotoxicity.

  20. Tolerance of monocytes and macrophages in response to bacterial endotoxin.

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    Wiśnik, Ewelina; Pikus, Ewa; Duchnowicz, Piotr; Koter-Michalak, Maria

    2017-03-07

    Monocytes belong to myeloid effector cells, which constitute the first line of defense against pathogens, also called the nonspecific immune system and play an important role in the maintenance of tissue homeostasis. In response to stimulation, monocytes differentiate into macrophages capable of microorganism phagocytosis and secrete factors that play a key role in the regulation of immune responses. However excessive exposure of monocytes/macrophages to the lipopolysaccharide (LPS) of Gram negative bacteria leads to the acquisition of immune tolerance by these cells. Such state results from disruption of different biological processes, for example intracellular signaling pathways and is accompanied by a number of disease states (immune, inflammatory or neoplastic conditions). Regulation of monocytes/macrophages activity is controlled by miRNAs, which are involved in the modulation of immune tolerance acquired by these cells. Moreover, the tolerance to endotoxin is conditioned by the posttranscriptional processes and posttranslational epigenetic modifications leading to the impairment of normal immune response for example by alterations in the expression of many genes encoding immune signaling mediators. The aim of this paper is to provide an overview existing knowledge on the modulation of activity of monocytes/macrophages in response to bacterial endotoxin and impaired immune responses.

  1. Tolerance of monocytes and macrophages in response to bacterial endotoxin

    Directory of Open Access Journals (Sweden)

    Ewelina Wiśnik

    2017-03-01

    Full Text Available Monocytes belong to myeloid effector cells, which constitute the first line of defense against pathogens, also called the nonspecific immune system and play an important role in the maintenance of tissue homeostasis. In response to stimulation, monocytes differentiate into macrophages capable of microorganism phagocytosis and secrete factors that play a key role in the regulation of immune responses. However excessive exposure of monocytes/macrophages to the lipopolysaccharide (LPS of Gram negative bacteria leads to the acquisition of immune tolerance by these cells. Such state results from disruption of different biological processes, for example intracellular signaling pathways and is accompanied by a number of disease states (immune, inflammatory or neoplastic conditions. Regulation of monocytes/macrophages activity is controlled by miRNAs, which are involved in the modulation of immune tolerance acquired by these cells. Moreover, the tolerance to endotoxin is conditioned by the posttranscriptional processes and posttranslational epigenetic modifications leading to the impairment of normal immune response for example by alterations in the expression of many genes encoding immune signaling mediators. The aim of this paper is to provide an overview existing knowledge on the modulation of activity of monocytes/macrophages in response to bacterial endotoxin and impaired immune responses.

  2. Recent advances in biosensor based endotoxin detection.

    Science.gov (United States)

    Das, A P; Kumar, P S; Swain, S

    2014-01-15

    Endotoxins also referred to as pyrogens are chemically lipopolysaccharides habitually found in food, environment and clinical products of bacterial origin and are unavoidable ubiquitous microbiological contaminants. Pernicious issues of its contamination result in high mortality and severe morbidities. Standard traditional techniques are slow and cumbersome, highlighting the pressing need for evoking agile endotoxin detection system. The early and prompt detection of endotoxin assumes prime importance in health care, pharmacological and biomedical sectors. The unparalleled recognition abilities of LAL biosensors perched with remarkable sensitivity, high stability and reproducibility have bestowed it with persistent reliability and their possible fabrication for commercial applicability. This review paper entails an overview of various trends in current techniques available and other possible alternatives in biosensor based endotoxin detection together with its classification, epidemiological aspects, thrust areas demanding endotoxin control, commercially available detection sensors and a revolutionary unprecedented approach narrating the influence of omics for endotoxin detection. Copyright © 2013 Elsevier B.V. All rights reserved.

  3. Trajectory of inflammatory and microglial activation markers in the postnatal rabbit brain following intrauterine endotoxin exposure.

    Science.gov (United States)

    Zhang, Zhi; Jyoti, Amar; Balakrishnan, Bindu; Williams, Monica; Singh, Sarabdeep; Chugani, Diane C; Kannan, Sujatha

    2018-03-01

    Maternal infection is a risk factor for periventricular leukomalacia and cerebral palsy (CP) in neonates. We have previously demonstrated hypomyelination and motor deficits in newborn rabbits, as seen in patients with cerebral palsy, following maternal intrauterine endotoxin administration. This was associated with increased microglial activation, primarily involving the periventricular region (PVR). In this study we hypothesized that maternal intrauterine inflammation leads to a pro-inflammatory environment in the PVR that is associated with microglial activation in the first 2 postnatal weeks. Timed pregnant New Zealand white rabbits underwent laparotomy on gestational day 28 (G28). They were randomly divided to receive lipopolysaccharide (LPS; 20μg/kg in 1mL saline) (Endotoxin group) or saline (1mL) (control saline, CS group), administrated along the wall of the uterus. The PVR from the CS and Endotoxin kits were harvested at G29 (1day post-injury), postnatal day1 (PND1, 3day post-injury) and PND5 (7days post-injury) for real-time PCR, ELISA and immunohistochemistry. Kits from CS and Endotoxin groups underwent longitudinal MicroPET imaging, with [ 11 C]PK11195, a tracer for microglial activation. We found that intrauterine endotoxin exposure resulted in pro-inflammatory microglial activation in the PVR of rabbits in the first postnatal week. This was evidenced by increased TSPO (translocator protein) expression co-localized with microglia/macrophages in the PVR, and changes in the microglial morphology (ameboid soma and retracted processes). In addition, CD11b level significantly increased with a concomitant decline in the CD45 level in the PVR at G29 and PND1. There was a significant elevation of pro-inflammatory cytokines and iNOS, and decreased anti-inflammatory markers in the Endotoxin kits at G29, PND1 and PND5. Increased [ 11 C]PK11195 binding to the TSPO measured in vivo by PET imaging in the brain of Endotoxin kits was present up to PND14-17. Our

  4. Pro-Inflammatory Cytokines but Not Endotoxin-Related Parameters Associate with Disease Severity in Patients with NAFLD.

    Directory of Open Access Journals (Sweden)

    Johannie du Plessis

    Full Text Available Intestinal dysbiosis and elevated lipopolysaccharides (LPS levels have been implicated in the development of obesity, insulin resistance and non-alcoholic steatohepatitis (NASH. In order to determine if LPS levels are elevated in patients with NASH compared to patients with non-alcoholic fatty liver (NAFL and, if elevated LPS levels correlated with histological severity of non-alcoholic fatty liver disease (NAFLD we compared LPS, markers of LPS bioactivity and pro-inflammatory cytokines/chemokines in patients undergoing bariatric surgery. At the time of surgery a liver biopsy was taken allowing the stratification into well-delineated subgroups including: No NAFL/NAFL; NASH; NASH with fibrosis and NASH cirrhotics, using the NAFLD Activity Score (NAS. Anthropometric data and plasma were collected for assessment of LPS, lipopolysaccharide binding protein (LBP, soluble CD14 (sCD14, intestinal-type fatty acid binding protein (iFABP, Toll-like receptors 2 and 4 (TLR2, 4 and a panel of cytokines/chemokines. Similar analysis was performed on plasma from a cohort of healthy controls. Our data indicate elevated levels of LPS, LBP, sCD14, iFABP and TLR2,4 in obese patients compared to healthy controls, however, these parameters remained unaltered within patients with limited liver disease (NAFL compared to NASH/NASH with fibrosis subgroups. Hierarchic cluster analysis using endotoxin-related parameters failed to discriminate between lean controls, NAFLD. While similar cluster analysis implementing inflammation-related parameters clearly distinguished lean controls, NALFD subgroups and NASH cirrhotics. In addition, LPS levels was not associated with disease severity while TNFα, IL8, and CCL3 featured a clear correlation with transaminase levels and the histological severity of NALFD. In conclusion our data indicate a stronger correlation for circulating inflammatory- rather than endotoxin-related parameters in progression of NAFLD and highlights the need

  5. Looped limulus anti-lipopolysaccharide derived peptide CLP-19 induces endotoxin tolerance involved inhibition of NF-κB activation.

    Science.gov (United States)

    Yang, Ya; Li, Di; Tian, Zhiqiang; Lv, Jun; Sun, Fengjun; Wang, Qian; Liu, Yao; Xia, Peiyuan

    2016-11-18

    Endotoxin tolerance (ET) is a complex protective mechanism against endotoxin shock. The looped CLP-19 peptide derived from Limulus anti-LPS peptide induced the ET phenomenon but the molecular mechanism has yet to be fully elucidated. Here, we confirmed that CLP-19 attenuated upon LPS stimulated pro-inflammatory factor secretion of TNF-α and IL-6 but increased anti-inflammatory factor production of IL-10 in dose- and time-dependent manners. CLP-19 also inhibited subsequent LPS stimulated expression of TLR4 on the cell membrane. Moreover, the CLP-19 inhibited degradation of the inhibitor of NF-κB (IκBα and IκBβ) and reduced LPS induced NF-κB activity, but not of effects on expression of MyD88 and TRAF-6. Finally CLP-19 significantly increased survival of lethal LPS shock mouse models with significantly less pathological injury to lung. These findings collectively suggest that CLP-19 induces ET phenomenon involved inhibition of NF-κB activation. In conclusion, this study has revealed a novel function of CLP-19 that appears to represent a potential therapeutic agent for clinical treatment of septic shock. Copyright © 2016 Elsevier Inc. All rights reserved.

  6. Directed evolution of an LBP/CD14 inhibitory peptide and its anti-endotoxin activity.

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    Li Fang

    Full Text Available BACKGROUND: LPS-binding protein (LBP and its ligand CD14 are located upstream of the signaling pathway for LPS-induced inflammation. Blocking LBP and CD14 binding might prevent LPS-induced inflammation. In previous studies, we obtained a peptide analog (MP12 for the LBP/CD14 binding site and showed that this peptide analog had anti-endotoxin activity. In this study, we used in vitro directed evolution for this peptide analog to improve its in vivo and in vitro anti-endotoxin activity. METHODS: We used error-prone PCR (ep-PCR and induced mutations in the C-terminus of LBP and attached the PCR products to T7 phages to establish a mutant phage display library. The positive clones that competed with LBP for CD14 binding was obtained by screening. We used both in vivo and in vitro experiments to compare the anti-endotoxin activities of a polypeptide designated P1 contained in a positive clone and MP12. RESULTS: 11 positive clones were obtained from among target phages. Sequencing showed that 9 positive clones had a threonine (T to methionine (M mutation in amino acid 287 of LBP. Compared to polypeptide MP12, polypeptide P1 significantly inhibited LPS-induced TNF-α expression and NF-κB activity in U937 cells (P<0.05. Compared to MP12, P1 significantly improved arterial oxygen pressure, an oxygenation index, and lung pathology scores in LPS-induced ARDS rats (P<0.05. CONCLUSION: By in vitro directed evolution of peptide analogs for the LBP/CD14 binding site, we established a new polypeptide (P1 with a threonine (T-to-methionine (M mutation in amino acid 287 of LBP. This polypeptide had high anti-endotoxin activity in vitro and in vivo, which suggested that amino acid 287 in the C-terminus of LBP may play an important role in LBP binding with CD14.

  7. Functional Toll-like receptor 4 expressed in lactotrophs mediates LPS-induced proliferation in experimental pituitary hyperplasia

    Energy Technology Data Exchange (ETDEWEB)

    Sabatino, María Eugenia; Sosa, Liliana del Valle; Petiti, Juan Pablo; Mukdsi, Jorge Humberto [Centro de Microscopía Electrónica, Instituto de Investigaciones en Ciencias de la Salud (INICSA-CONICET), Facultad de Ciencias Médicas, Universidad Nacional de Córdoba, Av. Enrique Barros y Enfermera Gordillo, Ciudad Universitaria, CP 5000, Córdoba (Argentina); Mascanfroni, Iván Darío; Pellizas, Claudia Gabriela [Centro de Investigaciones en Bioquímica Clínica e Inmunología (CIBICI-CONICET), Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, Av. Haya de la Torre y Medina Allende, Ciudad Universitaria, CP 5000, Córdoba (Argentina); Gutiérrez, Silvina; Torres, Alicia Inés [Centro de Microscopía Electrónica, Instituto de Investigaciones en Ciencias de la Salud (INICSA-CONICET), Facultad de Ciencias Médicas, Universidad Nacional de Córdoba, Av. Enrique Barros y Enfermera Gordillo, Ciudad Universitaria, CP 5000, Córdoba (Argentina); De Paul, Ana Lucía, E-mail: adepaul@cmefcm.uncor.edu [Centro de Microscopía Electrónica, Instituto de Investigaciones en Ciencias de la Salud (INICSA-CONICET), Facultad de Ciencias Médicas, Universidad Nacional de Córdoba, Av. Enrique Barros y Enfermera Gordillo, Ciudad Universitaria, CP 5000, Córdoba (Argentina)

    2013-11-15

    Toll like receptor 4 (TLR4) has been characterized for its ability to recognize bacterial endotoxin lipopolysaccharide (LPS). Considering that infections or inflammatory processes might contribute to the progression of pituitary tumors, we analyzed the TLR4 functional role by evaluating the LPS effect on lactotroph proliferation in primary cultures from experimental pituitary tumors, and examined the involvement of PI3K-Akt and NF-κB activation in this effect. In addition, the role of 17β-estradiol as a possible modulator of LPS-induced PRL cell proliferation was further investigated. In estrogen-induced hyperplasic pituitaries, LPS triggered lactotroph cell proliferation. However, endotoxin failed to increase the number of lactotrophs taking up BrdU in normal pituitaries. Moreover, incubation with anti-TLR4 antibody significantly reduced LPS-induced lactotroph proliferation, suggesting a functional role of this receptor. As a sign of TLR4 activation, an LPS challenge increased IL-6 release in normal and tumoral cells. By flow cytometry, TLR4 baseline expression was revealed at the plasma membrane of tumoral lactotrophs, without changes noted in the percentage of double PRL/TLR4 positive cells after LPS stimulus. Increases in TLR4 intracellular expression were detected as well as rises in CD14, p-Akt and NF-κB after an LPS challenge, as assessed by western blotting. The TLR4/PRL and PRL/NF-κB co-localization was also corroborated by immunofluorescence and the involvement of PI3K/Akt signaling in lactotroph proliferation and IL-6 release was revealed through the PI3K inhibitor Ly-294002. In addition, 17β-estradiol attenuated the LPS-evoked increase in tumoral lactotroph proliferation and IL-6 release. Collectively these results demonstrate the presence of functional TLR4 in lactotrophs from estrogen-induced hyperplasic pituitaries, which responded to the proliferative stimulation and IL-6 release induced by LPS through TLR4/CD14, with a contribution of the PI3K

  8. Effects of chitosan-nanoparticle-mediated tight junction opening on the oral absorption of endotoxins.

    Science.gov (United States)

    Sonaje, Kiran; Lin, Kun-Ju; Tseng, Michael T; Wey, Shiaw-Pyng; Su, Fang-Yi; Chuang, Er-Yuan; Hsu, Chia-Wei; Chen, Chiung-Tong; Sung, Hsing-Wen

    2011-11-01

    Recently, we reported a pH-responsive nanoparticle (NP) system shelled with chitosan (CS), which could effectively increase the oral absorption of insulin and produce a hypoglycemic effect, presumably due to the CS-mediated tight junction (TJ) opening. It has been often questioned whether CS can also enhance the absorption of endotoxins present in the small intestine. To address this concern, we studied the effect of CS NPs on the absorption of lipopolysaccharide (LPS), the most commonly found toxin in the gastrointestinal tract. To follow their biodistribution by the single-photon emission computed tomography/computed tomography, LPS and insulin were labeled with (99m)Tc-pertechnetate ((99m)Tc-LPS) and (123)iodine ((123)I-insulin), respectively. The (99m)Tc-LPS was ingested 1 h prior to the administration of the (123)I-insulin-loaded NPs to mimic the physiological conditions. The confocal and TEM micrographs show that the orally administered CS NPs were able to adhere and infiltrate through the mucus layer, approach the epithelial cells and mediate to open their TJs. The radioactivity associated with LPS was mainly restricted to the gastrointestinal tract, whereas (123)I-insulin started to appear in the urinary bladder at 3 h post administration. This observation indicates that the insulin-loaded in CS NPs can traverse across the intestinal epithelium and enter the systemic circulation, whereas LPS was unable to do so, probably because of the charge repulsion between the anionic LPS in the form of micelles and the negatively charged mucus layer. Our in vivo toxicity study further confirms that the enhancement of paracellular permeation by CS NPs did not promote the absorption of LPS. These results suggest that CS NPs can be used as a safe carrier for oral delivery of protein drugs. Copyright © 2011 Elsevier Ltd. All rights reserved.

  9. Characterization of endotoxins on orthopaedic fixation screws, using physicochemical surface analyses.

    Science.gov (United States)

    França, Rodrigo; Alfa, Michelle; Olson, Nancy; Yahia, L'Hocine; Sacher, Edward

    2017-02-01

    The objective of this study was to determine if surface analysis techniques could be used to detect endotoxin on stainless steel malleolus screws. New malleolus screws were compared to ones that had been coated in purified lipopolysaccharide (LPS) or Artificial Test Soil (ATS) containing lipopolysaccharide. X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), and time-of flight secondary ion mass spectrometry (TOF-SIMS) were used to assess the fixation screws surface. Organic material was visualized on the LPS and ATS-LPS inoculated screws but not on the new unsoiled screws. This was further supported by the peaks observed at masses between 40 and 100 D in TOF-SIMS spectra of the LPS and ATS-LPS inoculated screws. After deconvolution of N1s high resolution XPS spectra, the LPS inoculated screws showed amide groups whereas the ATS-LPS inoculated screws showed predominantly nitroso groups (C-NO). Our data demonstrate that surface analysis can be used to detect organic residuals present on fixation screws. The XPS data confirmed that LPS reacted predominantly with positively charged surface metallic ions (Fe and Cr), whereas proteins reacted with the surface oxide layer of fixation screws, forming C-NO groups. The application of these surface analysis techniques will be helpful in determining if the reprocessing of such items results in an accumulation of organic material that might lead to aseptic loosening, when implanted. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:240-247, 2017. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.

  10. Immunostimulatory capacity of dental casting alloys on endotoxin responsiveness.

    Science.gov (United States)

    Rachmawati, Dessy; von Blomberg, B Mary E; Kleverlaan, Cornelis J; Scheper, Rik J; van Hoogstraten, Ingrid M W

    2017-05-01

    Oral metal exposure has been associated with systemic and local adverse reactions, probably due to elemental release from the alloys. Although supraphysiological concentrations of salts from dentally applied metals can activate innate cells through TLR4 (Ni, Co, Pd) and TLR3 (Au), whether direct exposure to solid alloys can also trigger innate immune reactivity is still unknown. The purpose of this in vitro study was to determine whether dental cast alloy specimens can activate innate cells and influence their responsiveness to bacterial endotoxin. Human monocyte-derived dendritic cells (MoDC) and THP-1 cells were cultured on top of different alloy specimens (Ni-Cr, Co-Cr, Pd-Cu, Pd-Ag, Ti-6Al-4V, amalgam, gold, and stainless steel) or in alloy-exposed culture medium with or without endotoxin (lipopolysaccharide [LPS]; Escherichia coli 055:B5). Interleukin-8 (IL-8) production was used as the parameter for innate stimulation and evaluated by enzyme-linked immunosorbent assay after 24 hours of culture. The statistical significance of the effects of various casting alloys on the secretion of IL-8 was analyzed by using the nonparametric Wilcoxon rank sum test (α=.05). Dental cast alloys induced IL-8 production in MoDC and THP-1 cells, with Au and Pd-Cu providing the strongest stimulation. The alloy-exposed culture media tested contained sufficient stimulatory metal ions to induce detectable IL-8 production in THP-1 cells, except for the Ni-Cr and stainless steel exposed media. Au and Pd-Cu alloys were also most effective in potentiating LPS responsiveness as measured by IL-8 production. Using an in vitro culture system to expose MoDC and THP-1 cells to different alloy specimens this study showed that contact with the solid alloys, in particular when they contain Pd or Au, can trigger innate immune responses and augment responsiveness to bacterial endotoxin. Copyright © 2016 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All

  11. Compound list: LPS [Open TG-GATEs

    Lifescience Database Archive (English)

    Full Text Available LPS LPS 00A07 ftp://ftp.biosciencedbc.jp/archive/open-tggates/LATEST/Human/in_vitro/LPS.Human.in_vitro....Liver.zip ftp://ftp.biosciencedbc.jp/archive/open-tggates/LATEST/Rat/in_vitro/LPS.Rat.in_vitro

  12. Novel integrated and portable endotoxin detection system based on an electrochemical biosensor.

    Science.gov (United States)

    Zuzuarregui, Ana; Souto, David; Pérez-Lorenzo, Eva; Arizti, Fernando; Sánchez-Gómez, Susana; Martínez de Tejada, Guillermo; Brandenburg, Klaus; Arana, Sergio; Mujika, Maite

    2015-01-21

    This paper describes the design, implementation and validation of a sensitive and integral technology solution for endotoxin detection. The unified and portable platform is based on the electrochemical detection of endotoxins using a synthetic peptide immobilized on a thin-film biosensor. The work covers the fabrication of an optimized sensor, the biofunctionalization protocol and the design and implementation of the measuring and signalling elements (a microfluidic chamber and a portable potentiostat-galvanostat), framed ad hoc for this specific application. The use of thin-film technologies to fabricate the biosensing device and the application of simple immobilization and detection methods enable a rapid, easy and sensitive technique for in situ and real time LPS detection.

  13. Increased intestinal permeability correlates with sigmoid mucosa alpha-synuclein staining and endotoxin exposure markers in early Parkinson's disease.

    Directory of Open Access Journals (Sweden)

    Christopher B Forsyth

    Full Text Available Parkinson's disease (PD is the second most common neurodegenerative disorder of aging. The pathological hallmark of PD is neuronal inclusions termed Lewy bodies whose main component is alpha-synuclein protein. The finding of these Lewy bodies in the intestinal enteric nerves led to the hypothesis that the intestine might be an early site of PD disease in response to an environmental toxin or pathogen. One potential mechanism for environmental toxin(s and proinflammatory luminal products to gain access to mucosal neuronal tissue and promote oxidative stress is compromised intestinal barrier integrity. However, the role of intestinal permeability in PD has never been tested. We hypothesized that PD subjects might exhibit increased intestinal permeability to proinflammatory bacterial products in the intestine. To test our hypothesis we evaluated intestinal permeability in subjects newly diagnosed with PD and compared their values to healthy subjects. In addition, we obtained intestinal biopsies from both groups and used immunohistochemistry to assess bacterial translocation, nitrotyrosine (oxidative stress, and alpha-synuclein. We also evaluated serum markers of endotoxin exposure including LPS binding protein (LBP. Our data show that our PD subjects exhibit significantly greater intestinal permeability (gut leakiness than controls. In addition, this intestinal hyperpermeability significantly correlated with increased intestinal mucosa staining for E. coli bacteria, nitrotyrosine, and alpha-synuclein as well as serum LBP levels in PD subjects. These data represent not only the first demonstration of abnormal intestinal permeability in PD subjects but also the first correlation of increased intestinal permeability in PD with intestinal alpha-synuclein (the hallmark of PD, as well as staining for gram negative bacteria and tissue oxidative stress. Our study may thus shed new light on PD pathogenesis as well as provide a new method for earlier

  14. Effect of LPS treatment on tyrosine hydroxylase expression and Parkinson-like behaviors.

    Science.gov (United States)

    Girard-Joyal, Olivier; Ismail, Nafissa

    2017-03-01

    Puberty is a critical period of development during which the brain undergoes reorganizing and remodeling. Exposure to stress during this period is thought to interfere with normal brain development and increase susceptibility to mental illnesses. In female mice, pubertal exposure to lipopolysaccharide (LPS), a bacterial endotoxin, has been shown to alter sexual, anxiety-like, and depression-like behaviors and cognition in an enduring manner. However, the mechanisms underlying these effects remain unknown. The present study examined age and sex difference in tyrosine hydroxylase (TH) expression and dopamine-dependent and Parkinson-like behaviors following LPS treatment. The results show that LPS treatment during adulthood causes an enduring increase in TH expression in many of the brain regions examined. In contrast, there is no change in TH expression following LPS treatment during puberty. However, pubertal LPS treatment induces enduring behavioral deficits in tests of Parkinson-like behaviors, more so in male than in female mice. These results suggest that the low levels of TH following exposure to pubertal immune challenge may predispose mice to Parkinson-like behavior. These findings add to our understanding of stress and immune responses during puberty and their impact on mental health later in life. Copyright © 2016 Elsevier Inc. All rights reserved.

  15. Association of endotoxins and colon polyp: a case-control study.

    Science.gov (United States)

    Lee, Kang-Kon; Yum, Keun-Sang

    2012-09-01

    Endotoxins are known to be associated with the occurrence of various chronic diseases. This study was conducted to investigate the role of endotoxins in the pathogenesis of colon polyps through a case-control study. A total of 145 subjects (74 subjects in the polyp group and 71 subjects in the control group) had undergone a colonoscopy. Age, body mass index (BMI) and endotoxin levels were found to be significantly higher in the polyp group than in the control group. The endotoxin level was still significantly higher in the polyp group than in the control group, even after age and BMI had been adjusted (polyp group 0.108 ± 0.007 EU/mL, control group 0.049 ± 0.008 EU/mL, P endotoxin level significantly increased in accordance with the number of colon polyps (one-polyp group, 0.088 ± 0.059 EU/mL; two-polyp group, 0.097 ± 0.071 EU/mL; three-or-more-polyp group, 0.149 ± 0.223 EU/mL). The endotoxin levels also significantly increased in groups with tubular adenoma with high-grade dysplasia (hyperplastic polyp group, 0.109 ± 0.121 EU/mL; tubular adenoma with low grade dysplasia group, 0.103 ± 0.059 EU/mL; tubular adenoma with high grade dysplasia group, 2.915 ± 0.072 EU/mL). In conclusion, the serum level of endotoxins is quantitatively correlated with colon polyps.

  16. T4 Phage Tail Adhesin Gp12 Counteracts LPS-induced Inflammation In Vivo

    Directory of Open Access Journals (Sweden)

    Paulina Miernikiewicz

    2016-07-01

    Full Text Available Bacteriophages that infect Gram-negative bacteria often bind to the bacterial surface by interaction of specific proteins with lipopolysaccharide (LPS. Short tail fiber proteins (tail adhesin, gp12 mediate adsorption of T4-like bacteriophages to Escherichia coli, binding surface proteins or LPS. Produced as a recombined protein, gp12 retains its ability to bind LPS. Since LPS is able to exert a major impact on the immune response in animals and in humans, we have tested LPS-binding phage protein gp12 as a potential modulator of the LPS-induced immune response. We have produced tail adhesin gp12 in a bacterial expression system and confirmed its ability to form trimers and to bind lipopolysaccharide in vitro by dynamic light scattering. This product had no negative effect on mammalian cell proliferation in vitro. Further, no harmful effects of this protein were observed in mice. Thus, gp12 was used in combination with LPS in a murine model, and it decreased the inflammatory response to LPS in vivo, as assessed by serum levels of cytokines IL-1 alpha and IL-6 and by histopathological analysis of spleen, liver, kidney and lungs. Thus, in future studies gp12 may be considered as a potential tool for modulation and specifically for counteracting LPS-related physiological effects in vivo.

  17. Health effects due to endotoxin inhalation (review)

    Energy Technology Data Exchange (ETDEWEB)

    Liebers, V.; Raulf-Heimsoth, M.; Bruening, T. [Ruhr-University Bochum, BGFA, Research Institute of Occupational Medicine, German Social Accident Insurance, Bochum (Germany)

    2008-04-15

    Endotoxins are ubiquitous in the environment and represent important components of bioaerosols. High exposure occurs in rural environment and at several workplaces (e.g. waste collecting, textile industry etc.). Adverse effects on human health induced by inhalation of endotoxin are described in several studies. Up to now the endotoxin levels are mainly measured using the Limulus amoebocyte-lysate (LAL) assay. This assay is well established, but for a suitable characterization of bioaerosols more parameters are necessary. Additional information, e.g. concerning the pyrogenic activity of organic dust samples may be delivered by whole blood assay. Whereas on the one hand protection measures at workplaces are demanded to avoid lung function impairment due to endotoxin exposure, on the other hand a protective effect of exposure to microbial agents like endotoxins with regard to allergy development has been observed. On the cellular level toll-like receptor 4 (TLR4) and IL-1 receptor as well as surface molecules like CD14 have been shown to play a pivotal role in the endotoxin activation cascade. In this review we summarize the mechanism of endotoxin recognition and its manifold effects on human health. (orig.)

  18. Field Studies Measuring the aerosolization of Endotoxin ...

    Science.gov (United States)

    Endotoxin is a component of the cell walls of Gram-negative bacteria and is known to be present in biosolids. Endotoxins have been shown to be a potent stimulator of the innate immune response causing airway irritation and shortness of breath. Class B biosolids are routinely applied to agricultural lands in the US to enhance soil properties and can be used as an alternative to chemical fertilizers. This study investigated the aerosolized endotoxin produced during the land application of Class B biosolids from various wastewater treatment plants on agricultural land and a concrete surface at two sites in Colorado, USA. Aerosolized endotoxin was captured using HiVol sampler fitted with glass fiber filter, polycarbonate filter cassette (both open and closed), and BioSampler impinger air samplers. Endotoxins were also measured in the bulk biosolids to allow for correlating bulk biosolids concentrations with aerosol emission rates. Endotoxin concentrations in biosolids, impinger solutions, and filter extracts were determined using the kinetic Limulus amebocyte lysate assay. Aerosolized endotoxin concentration was detected from all sites with levels ranging from 0.5 to 642 EU/m3. The four types of sampling apparatus were compared and the HiVol and open-faced cassette samplers used produced higher TWA measurements (EU/m3) than the impinger and closed cassette samplers. Ambient wind speed at the sites was found to be the variable best describing the results wit

  19. Activation of innate immune genes in caprine blood leukocytes after systemic endotoxin challenge

    DEFF Research Database (Denmark)

    Salvesen, Øyvind; Reiten, Malin R; Heegaard, Peter M. H.

    2016-01-01

    , and blood samples were collected throughout the trial. Characteristic signs of acute sepsis, such as sickness behavior, fever, and leukopenia were observed within 1 h of LPS administration. A high-throughput longitudinal gene expression analysis of circulating leukocytes was performed, and genes associated...... in leukocytes in response to LPS supporting a link between the interferon system and defense against bacterial infections. The extrahepatic expression of APPs suggests that leukocytes contribute to synthesis of these proteins at the beginning of a systemic inflammation. Taken together, these findings provide...... insights into the dynamic regulation of innate immune genes, as well as raising new questions regarding the importance of ISGs and extrahepatic APPs in leukocytes after systemic endotoxin challenge....

  20. Endotoxins in urban air in Stockholm, Sweden

    Science.gov (United States)

    Nilsson, S.; Merritt, A. S.; Bellander, T.

    2011-01-01

    Endotoxins, i.e. components originating from the outer membrane in the cell wall of Gram-negative bacteria, activate the human immune system, which may result in airway symptoms such as shortness of breath and airway inflammation. Endotoxins are present in the environment, both outdoors and indoors, and stay airborne for a long time. In order to investigate the levels of endotoxins in urban air and the influence of traffic and meteorological factors, particles (PM 10 and PM 2.5) were collected at five sites in Stockholm, Sweden on four occasions per site between May and September 2009. Endotoxins were extracted from the filters and analysis was conducted with the Limulus Amebocyte Lysate (LAL)-assay. Endotoxins were present in urban air in Stockholm, albeit in low levels, and were similar to levels found in urban areas outside Sweden. To our knowledge, this is the northernmost location where endotoxins have been measured. The endotoxin levels found in PM 10 ranged from 0.020 to 0.107 EU m -3 with a geometric mean of 0.050 EU m -3 and the levels found in PM 2.5 ranged from 0.005 to 0.064 EU m -3 with a geometric mean of 0.015 EU m -3. No obvious effects of traffic or meteorological factors on endotoxin levels were observed, although a moderate correlation could be seen with soot. The small number of sampling sites is however a shortcoming of the present study. In future studies, more sites and sampling during all seasons would be preferable in order to get a better picture of the influence of different sources on endotoxin levels.

  1. THE SUSCEPTIBILITY OF MICE TO BACTERIAL ENDOTOXINS

    Science.gov (United States)

    Schaedler, Russell W.; Dubos, Rene J.

    1961-01-01

    Albino mice (Rockefeller NCS strain) raised and maintained free of ordinary bacterial pathogens, as well as of intestinal Escherichia coli and of Proteus bacilli, were found to be highly resistant to the lethal effect of bacterial endotoxins. When newborn mice of this NCS colony were nursed by foster mothers from another colony raised under ordinary conditions (SS colony from which the NCS colony was derived), they acquired the intestinal flora of the latter animals and became susceptible to the lethal effects of endotoxins. NCS adult mice could be rendered susceptible to the lethal effect of endotoxins by vaccination with heat killed Gram-negative bacilli. The susceptibility thus induced exhibited a certain degree of specificity for the bacterial strain used in vaccination. Although untreated NCS mice were resistant to the lethal effect of endotoxins, they proved exquisitively susceptible to the infection-enhancing effect of these materials. For example, 1 µg. or less of endotoxin was found sufficient to help establish a rapidly fatal septicemia with Staphylococcus aureus. Small amounts of endotoxin (1 µg. or less), administered alone, caused a marked but transient loss of weight. Vaccination with heat-killed Gram-negative bacilli or with killed BCG increased the resistance of NCS mice to the infection-enhancing effect of small amounts of endotoxin. This protective effect exhibited a certain degree of specificity for the bacterial strain from which the toxin used in the infection-enhancing test was derived. These various findings can be explained by assuming that the pathological effects of endotoxins involve at least two unrelated mechanisms; (a) a primary toxicity illustrated in this study by the loss of weight and enhancement of infection resulting from the injection of small doses of toxin; (b) an immunological reaction with lethal consequences which became manifest only in animals sensitized to the endotoxin by prior exposure to Gram-negative bacilli. PMID

  2. Tissue protein nitration and peripheral blood endotoxin activity are indicative of the severity of systemic organ compromise in naturally-occurring clinical cases of bacterial mastitis in Holstein dairy cows

    Science.gov (United States)

    The objective of this survey study was to determine a relationship between the intensity of tissue protein tyrosine nitration measured in samples of mammary gland, liver, pancreas and lung compared to estimated blood endotoxin (LPS) activity. Blood was collected from nine multiparous Holstein cows...

  3. Kinetics of Hydrothermal Inactivation of Endotoxins

    OpenAIRE

    Li, Lixiong; Wilbur, Chris L.; Mintz, Kathryn L.

    2011-01-01

    A kinetic model was established for the inactivation of endotoxins in water at temperatures ranging from 210°C to 270°C and a pressure of 6.2 × 106 Pa. Data were generated using a bench scale continuous-flow reactor system to process feed water spiked with endotoxin standard (Escherichia coli O113:H10). Product water samples were collected and quantified by the Limulus amebocyte lysate assay. At 250°C, 5-log endotoxin inactivation was achieved in about 1 s of exposure, followed by a lower ina...

  4. Endotoxin levels and contribution factors of endotoxins in resident, school, and office environments - A review

    Science.gov (United States)

    Salonen, Heidi; Duchaine, Caroline; Létourneau, Valérie; Mazaheri, Mandana; Laitinen, Sirpa; Clifford, Sam; Mikkola, Raimo; Lappalainen, Sanna; Reijula, Kari; Morawska, Lidia

    2016-10-01

    As endotoxin exposure has known effects on human health, it is important to know the generally existing levels of endotoxins as well as their contributing factors. This work reviews current knowledge on the endotoxin loads in settled floor dust, concentrations of endotoxins in indoor air, and different environmental factors potentially affecting endotoxin levels. The literature review consists of peer-reviewed manuscripts located using Google and PubMed, with search terms based on individual words and combinations. References from relevant articles have also been searched. Analysis of the data showed that in residential, school, and office environments, the mean endotoxin loads in settled floor dust varied between 660 and 107,000 EU/m2, 2180 and 48,000 EU/m2, and 2700 and 12,890 EU/m2, respectively. Correspondingly, the mean endotoxin concentrations in indoor air varied between 0.04 and 1610 EU/m3 in residences, and 0.07 and 9.30 EU/m3 in schools and offices. There is strong scientific evidence indicating that age of houses (or housing unit year category), cleaning, farm or rural living, flooring materials (the presence of carpets), number of occupants, the presence of dogs or cats indoors, and relative humidity affect endotoxin loads in settled floor dust. The presence of pets (especially dogs) was extremely strongly associated with endotoxin concentrations in indoor air. However, as reviewed articles show inconsistency, additional studies on these and other possible predicting factors are needed.

  5. Neutralization of endotoxin in vitro and in vivo by Bac7(1-35), a proline-rich antibacterial peptide.

    Science.gov (United States)

    Ghiselli, Roberto; Giacometti, Andrea; Cirioni, Oscar; Circo, Raffaella; Mocchegiani, Federico; Skerlavaj, Barbara; D'Amato, Giuseppina; Scalise, Giorgio; Zanetti, Margherita; Saba, Vittorio

    2003-06-01

    Lipopolysaccharides (LPS), or endotoxins, are structural components of gram-negative bacteria implicated in the pathogenesis of septic shock. In this study the antiendotoxin activity of Bac7(1-35), a synthetic peptide based on the sequence of a proline-rich antibacterial peptide from bovine neutrophils, was investigated in vitro and in an experimental rat model of gram-negative septic shock. The ability of Bac7(1-35) to bind LPS from Escherichia coli O111:B4 was determined using a sensitive Limulus chromogenic assay. In the in vivo study, adult male Wistar rats were given an intraperitoneal injection of 1 x 10(9) colony-forming units of E. coli ATCC 25922. All animals were randomized to receive intraperitoneally 1 mg/kg Bac7(1-35), or isotonic sodium chloride solution (control group C1), 60 mg/kg of piperacillin and 1 mg/kg polymyxin B, 1 mg/kg of polymyxin B plus 60 mg/kg of piperacillin, and 1 mg/kg of Bac7(1-35) plus 60 mg/kg of piperacillin. Each group included 15 animals. Bac7(1-35) was found to completely inhibit the LPS procoagulant activity at approximately 10 microM peptide concentration, as determined by in vitro LAL chromogenic assay. Treatment with Bac7(1-35) resulted in significant decrease in plasma endotoxin levels and lethality rates compared with saline injected control animals. No statistically significant differences were noted between Bac7(1-35) and polymyxin B in reducing all variables measured. These results provide evidence for the ability of Bac7(1-35) to effectively bind LPS and protect animals from lethal effects of this molecule, and point to its potential use for the treatment of endotoxin-induced septic shock.

  6. 21 CFR 866.3210 - Endotoxin assay.

    Science.gov (United States)

    2010-04-01

    ... DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3210 Endotoxin assay. (a... intended for use in conjunction with other laboratory findings and clinical assessment of the patient to...

  7. Endotoxins: relationship between structure, function, and activity.

    Science.gov (United States)

    Brandenburg, K; Schromm, A B; Gutsmann, T

    2010-01-01

    Endotoxins as amphiphilic components of the outer layer of the outer membrane of Gram-negative bacteria exert their immunostimulatory activity after release from bacterial cells. Thus, the characterization of the physicochemical properties of this glycolipid in physiological fluids is of utmost importance for an understanding of cell activation processes. Here, the essential physicochemical parameters describing endotoxins such as critical micellar concentration, acyl chain fluidity, intramolecular conformation, supramolecular structures, and size as well as morphology of the aggregates are discussed and assessed with respect to their importance for an understanding of the interaction mechanisms with immunorelevant cells. The reviewed data clearly indicate that knowledge of these parameters is essential for understanding the bioactivity of not only endotoxins, but also endotoxin-like amphiphiles.

  8. MODIFICATION OF HOST RESPONSES TO BACTERIAL ENDOTOXINS

    Science.gov (United States)

    Watson, Dennis W.; Kim, Yoon Berm

    1963-01-01

    Evidence is presented suggesting that the apparent non-specificity of pyrogenic tolerance observed with Gram-negative bacterial endotoxins is due to related antigenic determinants associated with the macromolecular toxins. This is based on results obtained in rabbits from pyrogenic cross-tolerance tests with selected endotoxins. In these tests, purified endotoxins from Escherichia coli (COO8) and Chromobacterium violaceum (CV) gave results one might expect with non-reciprocal cross-reacting antigens in classical immune systems. Additional evidence for an immune mechanism in tolerance is suggested by the highly significant anamnestic response observed. Lipid A, a toxic derivative of the purified COO8 endotoxin, failed to induce pyrogenic tolerance against the parent toxin. These results are explained by assuming that endotoxins have two interdependent activities associated with different portions of the macromolecule; one is assumed to be responsible for the primary toxicity, and the other is involved in secondary toxicity. The latter is dependent on the hypersensitive state of the host. Additional evidence for the role of hypersensitivity in secondary toxicity is based on the observation that adult rabbits are highly sensitive to the pyrogenic, lethal, and skin-reacting activities of endotoxin in contrast to young animals which are more resistant to all of these attributes of toxicity. In adults, the host responses to pyrogenicity, lethality, and skin reactivity could be partially inhibited by the early exposure of the animals to massive doses of endotoxin equivalent to a LD50. The pyrogenic tolerance shown in these animals was specific indicating that the inhibition of the hypersusceptibility to endotoxin involved an immunological mechanism. A mechanism of endotoxin tolerance is proposed and discussed based on the induction of specific antibodies capable of assisting the RES in the clearance and destruction of endotoxin. It is suggested that the present

  9. Erythropoietin augments the cytokine response to acute endotoxin-induced inflammation in humans

    DEFF Research Database (Denmark)

    Hojman, Pernille; Taudorf, Sarah; Lundby, Carsten

    2009-01-01

    Recent studies have shown that erythropoietin (EPO) offers protection against ischemia, hemorrhagic shock and systemic inflammation in many tissues and it has been suggested that EPO has anti-inflammatory effects. With the aim of investigating the potential acute anti-inflammatory effects of EPO ...... with endotoxin, the levels of TNF-alpha and IL-6 were enhanced by 5- and 40-fold, respectively, whereas the endotoxin-induced increase in IL-10 response was not influenced by EPO. In contrast to our hypothesis, we find that EPO augments the acute inflammatory effect.......Recent studies have shown that erythropoietin (EPO) offers protection against ischemia, hemorrhagic shock and systemic inflammation in many tissues and it has been suggested that EPO has anti-inflammatory effects. With the aim of investigating the potential acute anti-inflammatory effects of EPO...... in a human in vivo model of acute systemic low-grade inflammation, we measured circulating inflammatory mediators after intravenous administration of Escherichia coli endotoxin (LPS) bolus injection (0.1 ng/kg of body weight) in young healthy male subjects. The subjects were divided into three groups...

  10. Β-hydroxymyristic acid as a chemical marker to detect endotoxins in dialysis water.

    Science.gov (United States)

    Mishra, Rupesh K; Robert-Peillard, Fabien; Ravier, Sylvain; Coulomb, Bruno; Boudenne, Jean-Luc

    2015-02-01

    An analytical chemical method has been developed for determination of β-hydroxymyristic acid (β-HMA), a component of lipopolysaccharides (LPSs/endotoxins) in dialysis water. In our investigation, the β-HMA component was used as a chemical marker for endotoxin presence in dialysis water because it is available in the molecular subunit (lipid A) and responsible for toxicity. It is the most abundant saturated fatty acid in that subunit. The developed method is based on fluorescence derivatization with 4-nitro-7-piperazino-2,1,3-benzoxadiazole (NBD-PZ). A high-performance liquid chromatographic separation of the β-HMA derivative was achieved using an octadecyl silica column in gradient elution. A wide dynamic range of β-HMA was tested and a calibration curve was constructed with accuracy of 90% and variability of less than 10%. The limits of detection and quantification obtained were 2 and 5μM, respectively. The developed method was applied to detect endotoxins in dialysis water by alkaline hydrolysis of LPS using NaOH (0.25M) at 60°C for 2h. After hydrolysis, free acid was detected as its NBD-PZ derivative using high-performance liquid chromatography/mass spectrometry (HPLC/MS). Good recovery rates ranging from 98 to 105% were obtained for β-HMA in dialysis water. Copyright © 2014 Elsevier Inc. All rights reserved.

  11. [Effect of paeonol on LPS-induced rat vascular endothelial cell adhesion reaction].

    Science.gov (United States)

    Chen, Jun-Jun; Dai, Min; Chen, Peng

    2013-03-01

    To observe the effect of Paeonol (Pae) on lipopolysaccharide (LPS) induced rat mononuclear cells (MCs) adhesion to vascular endothelial cells (VECs) and provide basis foundation for inflammatary mechanisms of Pae against atherosclerosis. Rat vascular endothelial cells were isolated with tissue predigested adherent method. LPS was used as stimulator to induce VEC injury. Serum containing Pae obtained from healthy rats which were given Pae in intragastric. RP-HPLC method was used for detecting the concentration of Pae in serum. MTT assay was used to determine the protective effect of Pae on injured VECs. Rose Bengal Staining was used to detect the effect of Pae on LPS-induced MCs adhesion to VECs. LPS induced rat MCs adhesion to VECs. The effect was the strongest when the concentration was 10 ng/mL and incubated with VECs for 5 h. Pae in concentration of 2. 5,5 and 10 microg/mL and incubated for 24 h could effectively inhibit the adhesion and improve the survival rate of LPS injured VECs significantly. LPS can damage VECs. Pae could protect VECs from LPS injury via inhibiting MCs adhesion to VECs and improving the VEC survival rate.

  12. Endotoxins in cotton: washing effects and size distribution

    Energy Technology Data Exchange (ETDEWEB)

    Olenchock, S.A.; Mull, J.C.; Jones, W.G.

    1983-01-01

    Endotoxin contamination was measured in washed and unwashed cottons from three distinct growing areas, California, Mississippi, and Texas. The data show differences in endotoxin contamination based upon the geographic source of the cotton. It is also shown that washing bulk cotton before the carding process results in lower endotoxin in the cotton dust. Washing conditions can affect the endotoxin levels, and all size fractions of the airborne dust contain quantifiable endotoxin contamination. Endotoxin analyses provide a simple and reliable method for monitoring the cleanliness of cotton or airborne cotton dusts.

  13. Comparison of acute endotoxin-induced lesions in A/J and C57BL/6J mice.

    Science.gov (United States)

    O'Malley, J; Matesic, L E; Zink, M C; Strandberg, J D; Mooney, M L; De Maio, A; Reeves, R H

    1998-01-01

    Resistance to the action of endotoxin varies among inbred strains of mice, indicating that a component of this resistance has a genetic basis. Different responses to endotoxin that are characteristic of individual inbred strains represent phenotypes that can be used to genetically map the response modifier genes. This study compares the acute histologic lesions in 8-week-old male A/J and C57BL/6J (B6) mice injected intraperitoneally with endotoxin of E. coli O265:B6 (15 mg/kg). Animals of both strains exhibited splenitis, splenic lymphoid hyperplasia, splenic lymphoid necrosis, and sequestration of neutrophils in the pulmonary alveoli. The B6 mice showed increased margination of white blood cells to the pulmonary vascular endothelium relative to A/J mice. A large number of degenerating neutrophils was observed in the liver sinusoids of most B6 animals, while this lesion was much less severe in A/J mice. This difference was quantified, demonstrating a highly significant difference in neutrophil infiltration in B6 mice relative to A/J mice. Analysis of this phenotype in F1 mice demonstrates that major genes encoding the trait are not X-linked, imprinted, or maternally inherited and do not show the codominant inheritance expected if Lps(d) were primarily responsible. The distinctive, quantitative nature of these differences provides a useful assay for mapping genes that modify endotoxin responsiveness using the AXB and BXA recombinant inbred (RI) strains derived from A/J and B6 mice.

  14. Effects of subacute ruminal acidosis challenges on fermentation and endotoxins in the rumen and hindgut of dairy cows.

    Science.gov (United States)

    Li, S; Khafipour, E; Krause, D O; Kroeker, A; Rodriguez-Lecompte, J C; Gozho, G N; Plaizier, J C

    2012-01-01

    The effects of a grain-based subacute ruminal acidosis (SARA) challenge (GBSC) and an alfalfa-pellet SARA challenge (APSC) on fermentation and endotoxins in the rumen and in the cecum, as well as on endotoxins in peripheral blood, were determined. Six nonlactating Holstein cows with cannulas in the rumen and cecum were used in the study. A 3×3 Latin square arrangement of treatments with 4-wk experimental periods was adopted. During the first 3 wk of each experimental period, all cows received a diet containing 70% forages [dry matter (DM) basis]. In wk 4 of each period, cows received 1 of the following 3 diets: the 70% forage diet fed during wk 1 to 3 (control), a diet in which 34% of the dietary DM was replaced with grain pellets made of 50% ground wheat and 50% ground barely (GBSC), or a diet in which 37% of dietary DM was replaced with pellets of ground alfalfa (APSC). Rumen pH was monitored continuously using indwelling pH probes, and rumen fluid, blood, cecal digesta, and fecal grab samples were collected immediately before feed delivery at 0900 h and at 6 h after feed delivery on d 3 and 5 of wk 4. The time for which rumen pH was below 5.6 was 56.4, 225.2, and 298.8 min/d for the control, APSC, and GBSC treatments, respectively. Compared with the control, SARA challenges resulted in similar reductions in cecal digesta pH, which were 7.07, 6.86, and 6.79 for the control, APSC, and GBSC treatments, respectively. Compared with the control, only GBSC increased starch content in cecal digesta, which averaged 2.8, 2.6, and 7.4% of DM for the control, APSC, and GBSC, respectively. Free lipopolysaccharide endotoxin (LPS) concentration in rumen fluid increased from 10,405 endotoxin units (EU)/mL in the control treatment to 30,715 and 168,391 EU/mL in APSC and GBSC, respectively. Additionally, GBSC increased the LPS concentration from 16,508 to 118,522 EU/g in wet cecal digesta, and from 12,832 to 93,154 EU/g in wet feces. The APSC treatment did not affect LPS

  15. Cathelicidin peptide sheep myeloid antimicrobial peptide-29 prevents endotoxin-induced mortality in rat models of septic shock.

    Science.gov (United States)

    Giacometti, Andrea; Cirioni, Oscar; Ghiselli, Roberto; Mocchegiani, Federico; D'Amato, Giuseppina; Circo, Raffaella; Orlando, Fiorenza; Skerlavaj, Barbara; Silvestri, Carmela; Saba, Vittorio; Zanetti, Margherita; Scalise, Giorgio

    2004-01-15

    The present study was designed to investigate the antiendotoxin activity and therapeutic efficacy of sheep myeloid antimicrobial peptide (SMAP)-29, a cathelicidin-derived peptide. The in vitro ability of SMAP-29 to bind LPS from Escherichia coli 0111:B4 was determined using a sensitive limulus chromogenic assay. Two rat models of septic shock were performed: (1) rats were injected intraperitoneally with 1 mg E. coli 0111:B4 LPS and (2) intraabdominal sepsis was induced via cecal ligation and single puncture. All animals were randomized to receive parenterally isotonic sodium chloride solution, 1 mg/kg SMAP-29, 1 mg/kg polymyxin B or 20 mg/kg imipenem. The main outcome measures were: abdominal exudate and plasma bacterial growth, plasma endotoxin and tumor necrosis factor-alpha concentrations, and lethality. The in vitro study showed that SMAP-29 completely inhibited the LPS procoagulant activity at approximately 10 microM peptide concentration. The in vivo experiments showed that all compounds reduced the lethality when compared with control animals. SMAP-29 achieved a substantial decrease in endotoxin and tumor necrosis factor-alpha plasma concentrations when compared with imipenem and saline treatment and exhibited a slightly lower antimicrobial activity than imipenem. No statistically significant differences were noted between SMAP-29 and polymyxin B. SMAP-29, because of its double antiendotoxin and antimicrobial activities, could be an interesting compound for septic shock treatment.

  16. Airborne endotoxin in woodworking (joinery) shops.

    Science.gov (United States)

    Harper, Martin; Andrew, Michael E

    2006-01-01

    Symptoms such as shortness of breath and cough have been noted in woodworking facilities even where wood dust itself is well-controlled. Suspicion has fallen on other possible contaminants in the workplace atmosphere, including bacterial endotoxin. A few studies have indicated potentially high endotoxin exposure with exposure to fresh wood in sawmills and in the production of fiberboard and chipboard, but fewer studies have been carried out on exposure to endotoxin in dry wood work, for example in joineries. A study of the endotoxin content of airborne wood dust samples from US woodworking facilities is presented, from the re-analysis of samples which previously had been taken to establish mass collection relationships between the IOM sampler, the closed-face 37 mm plastic cassette (CFC) sampler and the Button sampler. Endotoxin was strongly correlated with total dust, but the endotoxin content of a few fresh wood samples was found to be up to ten times higher per unit of wood dust than for dried-wood samples, and this difference was significant. No long-term time-weighted average sample exceeded the recommended limit value of 50 EU m(-3) (EU, endotoxin units)used in the Netherlands, although a number of the IOM samples came close (seven samples or 44% exceeded 20 EU m(-3)) and one short-term (48 minute) sample registered a high value of 73 EU m(-3). The geometric mean concentration from the IOM samples (11 EU m(-3)) is within the range of geometric means found from Australian joineries (3.7-60, combined: 24 EU m(-3)). In contrast, the corresponding values from the CFC (3.6 EU m(-3)), and the Button sampler (2.1 EU m(-3)) were much lower and no samples exceeded 20 EU m(-3). Endotoxin is likely only to be a significant problem in working with dried woods when associated with very high dust levels, where the wood dust itself is likely to be a cause for concern. The results from the few samples in this study where fresh wood was being worked were similar to results

  17. Endotoxin-induced basal respiration alterations of renal HK-2 cells: A sign of pathologic metabolism down-regulation

    Energy Technology Data Exchange (ETDEWEB)

    Quoilin, C., E-mail: cquoilin@ulg.ac.be [Laboratory of Biomedical Spectroscopy, Department of Physics, University of Liege, 4000 Liege (Belgium); Mouithys-Mickalad, A. [Center of Oxygen Research and Development, Department of Chemistry, University of Liege, 4000 Liege (Belgium); Duranteau, J. [Department of Anaesthesia and Surgical ICU, CHU Bicetre, University Paris XI Sud, 94275 Le Kremlin Bicetre (France); Gallez, B. [Biomedical Magnetic Resonance Group, Louvain Drug Research Institute, Universite catholique de Louvain, 1200 Brussels (Belgium); Hoebeke, M. [Laboratory of Biomedical Spectroscopy, Department of Physics, University of Liege, 4000 Liege (Belgium)

    2012-06-29

    Highlights: Black-Right-Pointing-Pointer A HK-2 cells model of inflammation-induced acute kidney injury. Black-Right-Pointing-Pointer Two oximetry methods: high resolution respirometry and ESR spectroscopy. Black-Right-Pointing-Pointer Oxygen consumption rates of renal cells decrease when treated with LPS. Black-Right-Pointing-Pointer Cells do not recover normal respiration when the LPS treatment is removed. Black-Right-Pointing-Pointer This basal respiration alteration is a sign of pathologic metabolism down-regulation. -- Abstract: To study the mechanism of oxygen regulation in inflammation-induced acute kidney injury, we investigate the effects of a bacterial endotoxin (lipopolysaccharide, LPS) on the basal respiration of proximal tubular epithelial cells (HK-2) both by high-resolution respirometry and electron spin resonance spectroscopy. These two complementary methods have shown that HK-2 cells exhibit a decreased oxygen consumption rate when treated with LPS. Surprisingly, this cellular respiration alteration persists even after the stress factor was removed. We suggested that this irreversible decrease in renal oxygen consumption after LPS challenge is related to a pathologic metabolic down-regulation such as a lack of oxygen utilization by cells.

  18. Mechanism of Hbγ-35-induced an increase in the activation of the human immune system by endotoxins.

    Science.gov (United States)

    Heinbockel, Lena; Palacios-Chaves, Leyre; Alexander, Christian; Rietschel, Ernst; Behrends, Jochen; Correa, Wilmar; Fukuoka, Satoshi; Gutsmann, Thomas; Ulmer, Artur J; Brandenburg, Klaus

    2015-04-01

    Endotoxins (LPS) are highly potent immune stimulatory molecules and are mainly known for triggering Gram-negative sepsis. However, besides their toxic effects, this stimulatory function may be advantageous, for example when used as an adjuvant during vaccination. Thus, there is always a narrow range between the useful wake-up of the immune system and its overwhelming reaction, which can lead to diseases like sepsis. This raises the question of which conformational properties are responsible for making the LPS aggregates more or less potent. As described previously, the size, type and form of LPS aggregates play a major role in their immune stimulatory activity. In this study we investigate the role of these parameters. On the one hand, we use a peptide (Pep19-2.5; Aspidasept) that causes a change of the LPS aggregate structure into a less toxic state; on the other hand, we use a potent immune stimulating peptide (Hbγ-35), leading to higher toxicity. We have found opposing effects on LPS aggregate conformations allowing a better understanding of the processes of immune stimulation. © The Author(s) 2014 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.

  19. Endotoxin Binding by Sevelamer: Potential Impact on Nutritional Status

    Directory of Open Access Journals (Sweden)

    Natsuki Kubotera

    2013-01-01

    Full Text Available Patients on hemodialysis (HD have a high burden of chronic inflammation induced associated with multiple comorbidities including poor nutritional status. Endotoxin (ET is a Gram-negative bacterial cell wall component and a potent stimulus for innate immune system activation leading to the transcription of proinflammatory cytokines (e.g., IL-1, IL-6, and TNFα that adversely affect protein metabolism and nutrition. Several cross-sectional observational studies have found that elevated serum ET concentrations in hemodialysis patients are associated with lower serum albumin, higher proinflammatory cytokine, and C-reactive protein concentrations. Possible sources of ET in the systemic circulation are bacterial translocation from the gastrointestinal tract and iron supplementation, potentially leading to intestinal bacterial overgrowth. Sevelamer is a nonabsorbable hydrogel approved for use as a phosphate binder in HD patients. Reductions in serum ET concentrations in hemodialysis patients have been observed with sevelamer therapy in observational studies and the few published interventional studies. Reduction of ET concentrations was associated with concomitant reductions in TNFα, IL-6, and CRP and improvement in serum albumin in the majority of these small studies. Additional studies are needed to evaluate the potential effects of sevelamer treatment on nutritional status in chronic kidney disease (CKD patients with elevated ET.

  20. Anti-inflammatory effect of lycopene on endotoxin-induced uveitis in rats.

    Science.gov (United States)

    Göncü, Tuğba; Oğuz, Elif; Sezen, Hatice; Koçarslan, Sezen; Oğuz, Halit; Akal, Ali; Adıbelli, Fatih Mehmet; Çakmak, Sevim; Aksoy, Nurten

    2016-01-01

    We evaluated the efficacy of lycopene, a dietary carotenoid and potent antioxidant, against ocular inflammation and oxidative stress in an experimental uveitis model. Endotoxin-induced uveitis (EIU) was induced in Sprague-Dawley rats by a single subcutaneous injection of 200 μg lipopolysaccharide (LPS). Induction of EIU was preceded by daily intraperitoneal injection of 10 mg/kg lycopene for three consecutive days (Lycopene + LPS group) or equivolume vehicle (Vehicle + LPS group). A positive control group received 1 mg/kg dexamethasone pretreatment (DEX + LPS), and a negative control group received daily vehicle injection but no LPS (Vehicle Control). Twenty-four hours after LPS or final vehicle administration, eyes were enucleated, and aqueous humor was collected for measurement of the number of infiltrating cells, total protein concentration, and levels of nitric oxide (NO), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and oxidative stress markers. Inflammatory response severity was compared among groups clinically and histopathologically. Infiltrating cell number, total protein concentration, and NO, TNF-α, and IL-6 levels were significantly elevated in the aqueous humor of Vehicle + LPS group rats compared to Vehicle Controls. Compared to the Vehicle + LPS group, lycopene pretreatment significantly reduced aqueous humor concentrations of oxidative stress markers, NO (0.29 ± 0.1 μM vs. 0.19 ± 0.1 μM, p=0.003), TNF-α (71.0 ± 22.3 ng/ml vs. 50.1 ± 2.1 ng/ml, p=0.043), and IL-6 (121.6 ± 3.0 pg/ml vs. 111.1 ± 5.6 pg/ml, p=0.008). Inflammatory score was also reduced (2.0 ± 0.0 vs. 0.4 ± 0.5, p=0.001). Lycopene reduced the infiltrating cell count and protein concentration, but differences did not reach significance. Most lycopene effects were equivalent to dexamethasone. Lycopene may aid in the clinical management of uveitis by suppressing inflammation and oxidative stress.

  1. [Therapeutic effect of a natural squamosamide derivative FLZ on Parkinson's disease model mice induced by LPS plus MPTP].

    Science.gov (United States)

    Yu, Ling-Hong; Wei, Huai-Ling; Bao, Xiu-Qi; Zhang, Dan; Sun, Hua

    2013-10-01

    The aim of this study is to investigate the protective effect of N-[2-(4-hydroxyphenyl)ethyl]-2-(2, 5-dimethoxyphenyl)-3-(3-methoxy-4-hydroxyphenyl)acrylamide (FLZ), a novel synthetic squamosamide cyclic derivative, against Parkinson's disease (PD) model mice induced by the inflammatory bacterial endotoxin, lipopolysaccharides (LPS) and the neurotoxin 1-methy-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP). C57/BL mice were ip injected LPS (5 mg x kg(-1)) once. One week following the LPS injection, mice received a subcutaneous injection of MPTP (25 mg x kg(-1)) once daily for 2 days. Eight weeks later, FLZ (25, 50 and 75 mg x kg(-1)) was orally administered to mice once daily for 60 days. The motor ability of the mice was evaluated by rod climbing test and footprint test. The dopamine (DA) levels in mouse striatum were determined by high performance liquid chromatography system. The tyrosine hydroxylase (TH)-positive cells were showed by immunohistochemical analysis. FLZ treatment significantly improved motor dysfunction of mice challenged by LPS plus MPTP. The increase of TH-positive cell numbers and elevation of DA levels may be contributed to the beneficial effects of FLZ on motor behavior. This study showed FLZ has significant therapeutic effect on LPS plus MPTP induced chronic PD model, which indicates its potential as a new candidate drug to treat PD.

  2. Endotoxins in surgical instruments of hip arthroplasty

    Directory of Open Access Journals (Sweden)

    Vania Regina Goveia

    2016-06-01

    Full Text Available Abstract OBJECTIVE To investigate endotoxins in sterilized surgical instruments used in hip arthroplasties. METHOD A descriptive exploratory study conducted in a public teaching hospital. Six types of surgical instruments were selected, namely: acetabulum rasp, femoral rasp, femoral head remover, chisel box, flexible bone reamer and femoral head test. The selection was based on the analysis of the difficulty in removing bone and blood residues during cleaning. The sample was made up of 60 surgical instruments, which were tested for endotoxins in three different stages. The EndosafeTM Gel-Clot LAL (Limulus Amebocyte Lysate method was used. RESULT There was consistent gel formation with positive analysis in eight instruments, corresponding to 13.3%, being four femoral rasps and four bone reamers. CONCLUSION Endotoxins in quantity ≥0.125 UE/mL were detected in 13.3% of the instruments tested.

  3. Anti-inflammatory and ultrastructural effects of Turkish propolis in a rat model of endotoxin-induced uveitis.

    Science.gov (United States)

    Ertürküner, Salime Pelin; Yaprak Saraç, Elif; Göçmez, Semil Selcen; Ekmekçi, Hakan; Öztürk, Zeynep Banu; Seçkin, İsmail; Sever, Özkan; Keskinbora, Kadircan

    2016-01-01

    Experimental animal models of acute uveitis, an inflammatory eye disease, can be established via endotoxin-induced inflammation. Propolis, a natural substance collected by honeybees from buds and tree exudates, has antioxidant, antibacterial, antiviral, and anti-inflammatory effects. We investigated the effects of propolis, obtained from the Sakarya province of Turkey, on endotoxin-induced uveitis using immunohistochemical, ultrastructural, and biochemical approaches. Male Wistar albino rats (n = 6/group) received intraperitoneal (ip) lipopolysaccharide (LPS) endotoxin (150 μg/kg) followed by aqueous extract of propolis (50 mg/kg ip) or vehicle; two additional groups received either saline (control) or propolis only. After 24 h, aqueous humor (AH) was collected from both eyes of each animal for analysis of tumor necrosis factor-α (TNF-α) and hypoxia-inducible factor-1α (HIF-1α). Right eyeballs were paraffin-embedded for immunohistochemical staining of nuclear factor κB (NF-κB)/p65 and left eyeballs were araldite-embedded for ultrastructural analysis. Treatment of LPS-induced uveitis with propolis significantly reduced ciliary body NF-κB/p65 immunoreactivity and AH levels of HIF-1α and TNF-α. Ultrastructural analysis showed fewer vacuoles and reduced mitochondrial degeneration in the retinal pigment epithelium, as compared to the uveitis group. The intercellular spaces of the inner nuclear layer and outer limiting membrane were comparable with those of the control group; no polymorphonuclear cells or stasis was observed in intravascular or extravascular spaces. This is the first report demonstrating an anti-inflammatory effect of Turkish propolis in a rat model of LPS-induced acute uveitis, suggesting a therapeutic potential of propolis for the treatment of inflammatory ophthalmic diseases.

  4. Differential Induction of the Toll-Like Receptor 4-MyD88-Dependent and -Independent Signaling Pathways by Endotoxins

    Science.gov (United States)

    Zughaier, Susu M.; Zimmer, Shanta M.; Datta, Anup; Carlson, Russell W.; Stephens, David S.

    2005-01-01

    The biological response to endotoxin mediated through the Toll-like receptor 4 (TLR4)-MD-2 receptor complex is directly related to lipid A structure or configuration. Endotoxin structure may also influence activation of the MyD88-dependent and -independent signaling pathways of TLR4. To address this possibility, human macrophage-like cell lines (THP-1, U937, and MM6) or murine macrophage RAW 264.7 cells were stimulated with picomolar concentrations of highly purified endotoxins. Harvested supernatants from previously stimulated cells were also used to stimulate RAW 264.7 or 23ScCr (TLR4-deficient) macrophages (i.e., indirect induction). Neisseria meningitidis lipooligosaccharide (LOS) was a potent direct inducer of the MyD88-dependent pathway molecules tumor necrosis factor alpha (TNF-α), interleukin-1β (IL-1β), monocyte chemoattractant protein 1 (MCP-1), macrophage inflammatory protein 3α (MIP-3α), and the MyD88-independent molecules beta interferon (IFN-β), nitric oxide, and IFN-γ-inducible protein 10 (IP-10). Escherichia coli 55:B5 and Vibrio cholerae lipopolysaccharides (LPSs) at the same pmole/ml lipid A concentrations induced comparable levels of TNF-α, IL-1β, and MIP-3α, but significantly less IFN-β, nitric oxide, and IP-10. In contrast, LPS from Salmonella enterica serovars Minnesota and Typhimurium induced amounts of IFN-β, nitric oxide, and IP-10 similar to meningococcal LOS but much less TNF-α and MIP-3α in time course and dose-response experiments. No MyD88-dependent or -independent response to endotoxin was seen in TLR4-deficient cell lines (C3H/HeJ and 23ScCr) and response was restored in TLR4-MD-2-transfected human embryonic kidney 293 cells. Blocking the MyD88-dependent pathway by DNMyD88 resulted in significant reduction of TNF-α release but did not influence nitric oxide release. IFN-β polyclonal antibody and IFN-α/β receptor 1 antibody significantly reduced nitric oxide release. N. meningitidis endotoxin was a potent agonist of

  5. The bovine acute phase response to endotoxin and Gram-negative bacteria

    DEFF Research Database (Denmark)

    Jacobsen, Stine

    exposure to LPS were employed: 1) intravenous bolus injection of Escherichia coli (E. coli) LPS inducing systemic inflammation (Papers I-III and Reports I and II), and 2) intramammary inoculation with E. coli inducing infection and local inflammation in the mammary gland (Paper IV). Systemic inflammation...... and inflammation of the mammary gland Six cows were included in this study on the basis of their ability to clear intramammary infection induced by intramammary inoculation of E. coli (mild, intermediate and severe infection). Milk and plasma serum amyloid A (SAA) concentrations were measured sequentially (at 0, 6...... aimed at containing infections, eliminating bacteria and restoring homeostasis. However, during infections or disease complexes in which LPS and/or Gram-negative bacteria persist, disease and pathological changes may result from the prolonged inflammatory response. For example, protracted LPS...

  6. Dose-dependent sickness behavior, abortion and inflammation induced by systemic LPS injection in pregnant mice.

    Science.gov (United States)

    Toyama, Rovana Paludo; Xikota, João Carlos; Schwarzbold, Marcelo L; Frode, Tania Silvia; Buss, Ziliani da Silva; Nunes, Jean Costa; Funchal, Gabriela Di Giunta; Nunes, Fernanda Costa; Walz, Roger; Pires, Maria Marlene de Souza

    2015-03-01

    Clinical and experimental evidences indicate that intrauterine inflammation during pregnancy is associated to brain damage. The objective of this study is to determine the effects of lipopolysaccharide in temperature, cytokine production and sickness behavior of pregnant dams. A single i.p. injection of lipopolysaccharide (LPS) (50, 150 or 300 µg/kg) was administered on E18. Controls received isotonic saline. Body temperature was controlled before and 3 h after injections. Animals' behavior was assessed by the OF test 3 h following treatment. Animals were sacrificed for leukocyte, IL-1β and TNF-α determination. Placental tissue and abortion were also examined. LPS administration elicited hypothermia. Abortion was observed in LPS 150 and 300 µg/kg. Leukocyte levels were significantly lower with LPS 300 µg/kg than in controls. LPS induced dose-dependent impairment in animals' locomotion. IL-1β serum and amniotic fluid were higher than the saline, and TNF-α serum and amniotic fluid increased when compared to controls. Placental histopathologic abnormality was not found. LPS induces dose-dependent sickness behavior and hypothermia in pregnant mice. Our findings suggest that the presence of inflammation may be a causative factor for premature labor and that Escherichia coli antigens modify the concentration of pro-inflammatory agents in circulatory system and intra-uterine environment.

  7. Endotoxin induces a delayed loss of TH-IR neurons in substantia nigra and motor behavioral deficits.

    Science.gov (United States)

    Liu, Yuxin; Qin, Liya; Wilson, Belinda; Wu, Xuefei; Qian, Li; Granholm, Ann-Charlotte; Crews, Fulton T; Hong, Jau-Shyong

    2008-09-01

    We have previously reported that a single injection of endotoxin, lipopolysaccharide (LPS, 5mg/kg, i.p.), causes a delayed and progressive loss of TH-IR neurons in the substantia nigra (SN) in C57BL/six male mice. In this study, we determined sex differences and behavioral deficits accompanying the loss of TH-IR neurons in response to peripheral LPS injection. A single injection of LPS (5mg/kg, i.p.) failed to produce any loss of TH-IR neurons in the SN of female mice over a 12-month period. To determine if multiple-injections were required, female mice received five injections of LPS (5mg/kg, i.p.) at either weekly or monthly intervals. Behavioral motor ability and TH-IR neuronal loss were determined after the first injection of LPS. We found significant differences in both behavioral activities and neuronal loss between these two injection paradigms. Between 7 and 20 months after the first injection of LPS, progressive behavioral changes, measured by rotor-rod and open-field activities, and neuronal loss in SN were observed in monthly injected, but not in weekly injected mice. In addition, reduced rotor-rod ability in monthly injected mice were restored following treatment of l-dopa/carbidopa (30 mg/3mg/kg), i.p.). Approximately 40 and 50% loss of TH-IR neurons at 9 and 20 months, respectively, was observed after exposure to LPS, suggesting that the behavioral deficit is related to loss of dopamine function in the nigra-striatal pathway. More intense immuno-staining of alpha-synuclein and inflammatory markers were detected in brain sections exposed to LPS. In conclusion, these results show that multi-LPS monthly injections can induce a delayed and progressive loss of TH-IR neurons and motor deficits which resemble the progressive nature of Parkinson's disease. Further, the present study reveals a clear sex difference: female mice are more resistant to LPS than male mice. Repeated monthly LPS injections are required to cause both motor behavioral deficits and DA

  8. Protective effects of bilberry ( Vaccinium myrtillus L.) extract against endotoxin-induced uveitis in mice.

    Science.gov (United States)

    Yao, Nan; Lan, Fang; He, Rong-Rong; Kurihara, Hiroshi

    2010-04-28

    Endotoxin-induced uveitis (EIU), a useful animal model of ocular inflammation, is induced by injection of lipopolysacharide (LPS). These experiments showed that the nitric oxide (NO) level significantly increased in the whole eye homogenate of BALB/C mice 24 h after footpad injection of LPS at a dosage of 100 mg/mouse. However, the elevated NO level was significantly reduced by oral administration of bilberry extract (containing 42.04% anthocyanins) at dosages of 50, 100, and 200 mg/kg/day for 5 days before the LPS injection. In addition, bilberry extract decreased malondialdehyde (MDA) level and increased oxygen radical absorbance capacity (ORAC) level, glutathione (GSH) level, vitamin C level, and total superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities. Moreover, bilberry extract increased expression of copper/zinc superoxide dismutase (CuZnSOD), manganese superoxide dismutase (MnSOD), and GPx mRNA. Taken together, bilberry extract showed protective effects against EIU, whereas the effects of bilberry extract (100 and 200 mg/kg/day, 5 days) were dose-dependent. In conclusion, these results provide new evidence to elucidate the beneficial effects of bilberry extract on eye health.

  9. Development of Endotoxin Tolerance Does Not Influence the Response to a Challenge with the Mucosal Live-Attenuated Influenza Vaccine in Humans In Vivo

    Directory of Open Access Journals (Sweden)

    Rebecca M. Koch

    2017-12-01

    Full Text Available IntroductionThe effects of bacterial infections on the response to subsequent viral infections are largely unknown. This is important to elucidate to increase insight into the pathophysiology of bacterial and viral co-infections, and to assess whether bacterial infections may influence the course of viral infections.MethodsHealthy male subjects received either bacterial endotoxin [Escherichia coli-derived lipopolysaccharide (LPS, 2 ng/kg, n = 15] or placebo (n = 15 intravenously, followed by intranasal Fluenz (live-attenuated influenza vaccine 1 week later.ResultsLPS administration resulted in increased plasma cytokine levels and development of endotoxin tolerance in vivo and ex vivo, illustrated by attenuated cytokine production upon rechallenge with LPS. Following Fluenz administration, infectivity for the Fluenz A/B strains was similar between the LPS–Fluenz and placebo–Fluenz groups (13/15 subjects in both groups. Also, the Fluenz-induced increase in temperature and IL-6, G-CSF and IP-10 concentrations in nasal wash were similar between both groups.ConclusionWhile endotoxemia profoundly attenuates the immune response upon a second LPS challenge, it does not influence the Fluenz-induced immune response. These results suggest immune suppression after bacterial infection does not alter the response to a subsequent viral infection.

  10. Exposure to airborne endotoxin in Italian greenhouses: environmental analyses.

    Science.gov (United States)

    Gioffrè, Angela; Marramao, Antonella; DI Gesu, Ignazio; Samele, Pasquale; Paba, Emilia; Marcelloni, Anna; Chiominto, Alessandra; Iavicoli, Sergio

    2017-10-19

    The peculiar characteristics of the greenhouses as confined spaces, microclimate and poor air exchange with the outside environment, encourage the development of a large number of biological agents. Endotoxin, is probably a major causative agent of occupational health problems. The objective of this study was to measure the concentrations of airborne endotoxin in greenhouses with different cultures. The influence of microclimate was studied in correlation with endotoxin levels and type of cultured vegetables. The data indicate that workers employed greenhouses are exposed to low levels of inhalable endotoxins; endotoxin concentrations do not correlate with the temperature and relative humidity values. A strong correlation between the leaf size and endotoxin concentration was observed. The mean concentration of endotoxins in the air of greenhouses is relatively low, however, there could be peaks of exposure during harvesting and eradication of broadleaf plants.

  11. Allantoin as a solid phase adsorbent for removing endotoxins.

    Science.gov (United States)

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

    2013-10-04

    In this study we present a simple and robust method for removing endotoxins from protein solutions by using crystals of the small-molecule compound 2,5-dioxo-4-imidazolidinyl urea (allantoin) as a solid phase adsorbent. Allantoin crystalline powder is added to a protein solution at supersaturated concentrations, endotoxins bind and undissolved allantoin crystals with bound endotoxins are removed by filtration or centrifugation. This method removes an average of 99.98% endotoxin for 20 test proteins. The average protein recovery is ∼80%. Endotoxin binding is largely independent of pH, conductivity, reducing agent and various organic solvents. This is consistent with a hydrogen-bond based binding mechanism. Allantoin does not affect protein activity and stability, and the use of allantoin as a solid phase adsorbent provides better endotoxin removal than anion exchange, polymixin affinity and biological affinity methods for endotoxin clearance. Copyright © 2013 Elsevier B.V. All rights reserved.

  12. [Effect of inhalation of aerosolized unfractioned heparin on peri alveolar coagulability and inflammatory response in endotoxin induced acute lung injury rat model].

    Science.gov (United States)

    Wang, Zong-Yu; Yang, Ba-Xian; Zhu, Xi; Wu, Sheng-Nan

    2011-04-01

    To observe the local changes in alveoli in intravenous endotoxin induced acute lung injury (ALI) rat model after inhalation of aerosolized unfractioned heparin (UFH), and to observe its effects on coagulability, fibrinolysis and inflammatory response. Eighty seven male Wistar rats were divided into groups according to table of random number: sham, model, heparin therapy (HT) and heparin prophylaxis (HP). Endotoxin induced ALI model was reproduced by intravenous administration of lipopolysaccharide (LPS). Rats in HP and HT groups received aerosolized UFH before and after injection with LPS respectively, while rats in both sham and model groups inhaled aerosolized normal saline. Rats in each group were respectively sacrificed at 6, 12 and 24 hours after intravenous administration of LPS, and bronchoalveolar lavage fluid (BALF) was collected. Enzyme linked immunosorbent assay was used to measure the level of thrombin antithrombin (TAT), tissue type plasminogen activator (t-PA), urokinase type plasminogen activator (u-PA), plasminogen activator inhibitor 1 (PAI-1), tumor necrosis factor α (TNF-α), interleukin 6 (IL-6) in BALF. At 6 hours after injury, the level of TAT (μg/L) in model group (3.346±0.585) was highest, that in HT group (2.764±0.100) was higher, and that in HP group (2.564±0.216) was lowest in BALF (all Pinhalation of aerosolized UFH resulted in lowering local coagulability, alleviating fibrinolytic depression, improving fibrinolysis , and attenuating inflammation in endotoxin induced ALI rat model. More prominent results will be obtained when it was use as a prophylactic measure. The optimal time of usage is 6 hours after endotoxin injection.

  13. Kinetics of Hydrothermal Inactivation of Endotoxins

    Science.gov (United States)

    Li, Lixiong; Wilbur, Chris L.; Mintz, Kathryn L.

    2011-01-01

    A kinetic model was established for the inactivation of endotoxins in water at temperatures ranging from 210°C to 270°C and a pressure of 6.2 × 106 Pa. Data were generated using a bench scale continuous-flow reactor system to process feed water spiked with endotoxin standard (Escherichia coli O113:H10). Product water samples were collected and quantified by the Limulus amebocyte lysate assay. At 250°C, 5-log endotoxin inactivation was achieved in about 1 s of exposure, followed by a lower inactivation rate. This non-log-linear pattern is similar to reported trends in microbial survival curves. Predictions and parameters of several non-log-linear models are presented. In the fast-reaction zone (3- to 5-log reduction), the Arrhenius rate constant fits well at temperatures ranging from 120°C to 250°C on the basis of data from this work and the literature. Both biphasic and modified Weibull models are comparable to account for both the high and low rates of inactivation in terms of prediction accuracy and the number of parameters used. A unified representation of thermal resistance curves for a 3-log reduction and a 3 D value associated with endotoxin inactivation and microbial survival, respectively, is presented. PMID:21193667

  14. Endotoxin exposure assessment - measurement and characterization

    NARCIS (Netherlands)

    Spaan, S.|info:eu-repo/dai/nl/275162230

    2008-01-01

    Endotoxin, one of the specific agents in organic dust that cause health effects, is part of the cell wall of Gram-negative bacteria. Gram-negative bacteria are present on for instance the surfaces of plants and in animal feces, and their occurrence, growth and amplification is influenced by many

  15. Apical, but not basolateral, endotoxin preincubation protects alveolar epithelial cells against hydrogen peroxide-induced loss of barrier function: the role of nitric oxide synthesis.

    Science.gov (United States)

    Rose, Frank; Guthmann, Bernd; Tenenbaum, Tobias; Fink, Ludger; Ghofrani, Ardeschir; Weissmann, Norbert; König, Peter; Ermert, Leander; Dahlem, Gabriele; Haenze, Joerg; Kummer, Wolfgang; Seeger, Werner; Grimminger, Friedrich

    2002-08-01

    The influence of LPS preincubation on hydrogen peroxide (H(2)O(2))-induced loss of epithelial barrier function was investigated in rat alveolar epithelial type II cells (ATII). Both apical and basolateral H(2)O(2) administration caused a manyfold increase in transepithelial [(3)H]mannitol passage. Apical but not basolateral preincubation of ATII with LPS did not influence control barrier properties but fully abrogated the H(2)O(2)-induced leakage response. The effect of apical LPS was CD14 dependent and was accompanied by a strong up-regulation of NO synthase II mRNA and protein and NO release. Inhibition of NO by N(G)-monomethyl-L-arginine suppressed the LPS effect, whereas it was reproduced by exogenous application of gaseous NO or NO donor agents. Manipulation of the glutathione homeostasis (buthionine-(S,R)-sulfoximine) and the cGMP pathway (1H-(1,2,4)oxadiazolo[4,3-alpha]quinoxaline-1-one; zaprinast) did not interfere with the protective effect of LPS. Superoxide (O*(-)(2)) generation by ATII cells was reduced by exogenous NO and LPS preincubation. O*(-)(2) scavenging with exogenous superoxide dismutase, the intracellular superoxide dismutase analog Mn(III)tetrakis(4-benzoic acid) porphyrin, and the superoxide scavenger nitroblue tetrazolium and, in particular, hydroxyl radical scavenging with hydroxyl radical scavenger 1,3-dimethyl-thiourea inhibited the H(2)O(2)-induced epithelial leakage response. In conclusion, apical but not basolateral LPS preincubation of ATII cells provides strong protection against H(2)O(2)-induced transepithelial leakage, attributable to an up-regulation of epithelial NO synthesis. It is suggested that the LPS-induced NO formation is effective via interaction with reactive oxygen species, including superoxide and hydroxyl radicals. The polarized epithelial response to LPS may be part of the lung innate immune system, activated by inhaled endotoxin or under conditions of pneumonia.

  16. Efek ekstrak daun singkong (Manihot utilissima terhadap ekspresi COX-2 pada monosit yang dipapar LPS E.coli (The effect of Manihot utilissima extracts on COX-2 expression of monocytes induced by LPS E. coli

    Directory of Open Access Journals (Sweden)

    Zahara Meilawaty

    2013-12-01

    Full Text Available Background: Periodontal disease is a common and widespread disease in the community. Gram negative bacteria have a role inperiodontitis. These bacteria secrete a variety of products such as endotoxin lipopolysaccharide (LPS, which causes the occurrenceof inflammation or infection. The body defense responses are neutrophils and mononuclear cells (monocytes and macrophages. Inresponse to defense mechanism, the body will be expressed enzyme cyclooxygenase (COX which functions convert arachidonic acidto prostaglandins. Cassava leaf cells known to play a role in reducing inflammation, but the mechanism for inhibiting COX-2, is notknown. Purpose: The study was aimed to determine the effect of cassava leaf extract (Manihot utilissima on expression of enzyme COX-2 in monocytes which were exposed by LPS E. coli. Methods: This study was in vitro experimental studies with the design of posttestonly control group design. The sample was the cassava leaves extract (Manihot utilissima at concentration of 12.5 % and 25 %. Theexpression of COX-2 was determined by immunocytochemistry method. Isolated monocytes were incubated in cassava leaf extract, andthen exposed to LPS, after washing imunostaning procedure was performed using a monoclonal antibody (MAb anti-human COX-2.The research data was the number of monocytes that express COX-2. Results: Expression of COX-2 in the group cassava leaf extractwas higher than the group that induced by LPS E. coli only. Conclusion: Cassava leaf extract did not inhibit the expression of COX-2in monocytes which were exposed by LPS E. coli.Latar belakang: Penyakit periodontal merupakan penyakit umum dan tersebar luas di masyarakat. Bakteri yang banyak berperanpada periodontitis adalah Gram negatif. Bakteri ini mengeluarkan berbagai produk antara lain endotoksin lipopolisakarida (LPS yangmenyebabkan inflamasi atau infeksi. Respon pertahanan tubuh pertama adalah netrofil dan sel mononuklear (monosit dan makrofag.Pada respon

  17. Fractal-like kinetics of intracellular enzymatic reactions: a chemical framework of endotoxin tolerance and a possible non-specific contribution of macromolecular crowding to cross-tolerance.

    Science.gov (United States)

    Vasilescu, Catalin; Olteanu, Mircea; Flondor, Paul; Calin, George A

    2013-09-14

    The response to endotoxin (LPS), and subsequent signal transduction lead to the production of cytokines such as tumor necrosis factor-α (TNF-α) by innate immune cells. Cells or organisms pretreated with endotoxin enter into a transient state of hyporesponsiveness, referred to as endotoxin tolerance (ET) which represents a particular case of negative preconditioning. Despite recent progress in understanding the molecular basis of ET, there is no consensus yet on the primary mechanism responsible for ET and for the more complex cases of cross tolerance. In this study, we examined the consequences of the macromolecular crowding (MMC) and of fractal-like kinetics (FLK) of intracellular enzymatic reactions on the LPS signaling machinery. We hypothesized that this particular type of enzyme kinetics may explain the development of ET phenomenon. Our aim in the present study was to characterize the chemical kinetics framework in ET and determine whether fractal-like kinetics explains, at least in part, ET. We developed an ordinary differential equations (ODE) mathematical model that took into account the links between the MMC and the LPS signaling machinery leading to ET. We proposed that the intracellular fractal environment (MMC) contributes to ET and developed two mathematical models of enzyme kinetics: one based on Kopelman's fractal-like kinetics framework and the other based on Savageau's power law model. Kopelman's model provides a good image of the potential influence of a fractal intracellular environment (MMC) on ET. The Savageau power law model also partially explains ET. The computer simulations supported the hypothesis that MMC and FLK may play a role in ET. The model highlights the links between the organization of the intracellular environment, MMC and the LPS signaling machinery leading to ET. Our FLK-based model does not minimize the role of the numerous negative regulatory factors. It simply draws attention to the fact that macromolecular crowding can

  18. Edaravone protects endotoxin-induced liver injury by inhibiting apoptosis and reducing proinflammatory cytokines

    Energy Technology Data Exchange (ETDEWEB)

    Zong, L. [Second Military Medical University, Changhai Hospital, Department of Anesthesiology, Shanghai, China, Department of Anesthesiology, Changhai Hospital, Second Military Medical University, Shanghai (China); No. 82 Hospital of People' s Liberation Army, Department of Anesthesiology, Jiangsu, China, Department of Anesthesiology, No. 82 Hospital of People' s Liberation Army, Jiangsu (China); Yu, Q.H. [Second Military Medical University, Changhai Hospital, Department of Gastroenterology, Shanghai, China, Department of Gastroenterology, Changhai Hospital, Second Military Medical University, Shanghai (China); Du, Y.X. [No. 82 Hospital of People' s Liberation Army, Department of Anesthesiology, Jiangsu, China, Department of Anesthesiology, No. 82 Hospital of People' s Liberation Army, Jiangsu (China); Deng, X.M. [Second Military Medical University, Changhai Hospital, Department of Anesthesiology, Shanghai, China, Department of Anesthesiology, Changhai Hospital, Second Military Medical University, Shanghai (China)

    2014-03-03

    Studies have shown that edaravone may prevent liver injury. This study aimed to investigate the effects of edaravone on the liver injury induced by D-galactosamine (GalN) and lipopolysaccharide (LPS) in female BALB/c mice. Edaravone was injected into mice 30 min before and 4 h after GalN/LPS injection. The survival rate was determined within the first 24 h. Animals were killed 8 h after GalN/LPS injection, and liver injury was biochemically and histologically assessed. Hepatocyte apoptosis was measured by TUNEL staining; proinflammatory cytokines [tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6)] in the liver were assayed by ELISA; expression of caspase-8 and caspase-3 proteins was detected by Western blot assay; and caspase-3 activity was also determined. Results showed that GalN/LPS induced marked elevations in serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT). Edaravone significantly inhibited elevation of serum AST and ALT, accompanied by an improvement in histological findings. Edaravone lowered the levels of TNF-α and IL-6 and reduced the number of TUNEL-positive cells. In addition, 24 h after edaravone treatment, caspase-3 activity and mortality were reduced. Edaravone may effectively ameliorate GalN/LPS-induced liver injury in mice by reducing proinflammatory cytokines and inhibiting apoptosis.

  19. Improved detection limits of bacterial endotoxins using new type of planar interdigital sensors

    KAUST Repository

    Syaifudin, A. R Mohd

    2012-10-01

    New types of planar interdigital sensors were fabricated by photolithography and etching techniques on a Silicon/Silicon Dioxide (Si/SiO2) wafer (single side polished). The sensors were then coated with APTES (3-aminopropyltrietoxysilane) a cross linker used to bind Polymyxin B (PmB) molecules on electrodes surface. PmB is an antimicrobial peptide produced by the Gram-positive bacterium-Bacillus which has specific binding properties to Lipopolysaccharide (LPS). This paper will discuss the fabrication process, coating and immobilization procedures and analysis of sensors\\' performance based on Impedance Spectroscopy method. The sensor sensitivity was compared to standard ToxinSensor Chromogenic LAL Endotoxin Assay Kit for verification. © 2012 IEEE.

  20. Endotoxins in Environmental and Clinical Samples Assessed by GC-Tandem MS

    Science.gov (United States)

    Szponar, Bogumila

    Bacteria appeared on the Earth millions years before us and human evolution was triggered by the constant presence of pathogenic and symbiotic microorganisms in our surroundings. Interplay occurred between higher organism and microbial consortia residing in the host organs and on the epithelial surfaces; another natural space of bacteria-human interaction is the indoor environment where we spend the majority of our lifetime. Indoor microbial exposure affects our well-being and can result in respiratory symptoms, such as allergies and asthma, since both dead and live microorganisms and their cell constituents, including lipopolysaccharides (LPS, endotoxins), interact with our immune system. Thus, there is a demand for robust tools for qualitative and quantitative determination of the microbial communities that we are exposed to.

  1. Nutritional Supplement of Hatchery Eggshell Membrane Improves Poultry Performance and Provides Resistance against Endotoxin Stress.

    Directory of Open Access Journals (Sweden)

    S K Makkar

    Full Text Available Eggshells are significant part of hatchery waste which consist of calcium carbonate crust, membranes, and proteins and peptides of embryonic origins along with other entrapped contaminants including microbes. We hypothesized that using this product as a nutritional additive in poultry diet may confer better immunity to the chickens in the paradigm of mammalian milk that enhances immunity. Therefore, we investigated the effect of hatchery eggshell membranes (HESM as a short term feed supplement on growth performance and immunity of chickens under bacterial lipopolysaccharide (LPS challenged condition. Three studies were conducted to find the effect of HESM supplement on post hatch chickens. In the first study, the chickens were fed either a control diet or diets containing 0.5% whey protein or HESM as supplement and evaluated at 5 weeks of age using growth, hematology, clinical chemistry, plasma immunoglobulins, and corticosterone as variables. The second and third studies were done to compare the effects of LPS on control and HESM fed birds at 5 weeks of age following at 4 and 24 h of treatment where the HESM was also sterilized with ethanol to deplete bacterial factors. HESM supplement caused weight gain in 2 experiments and decreased blood corticosterone concentrations. While LPS caused a significant loss in body weight at 24 h following its administration, the HESM supplemented birds showed significantly less body weight loss compared with the control fed birds. The WBC, heterophil/lymphocyte ratio, and the levels of IgG were low in chickens fed diets with HESM supplement compared with control diet group. LPS challenge increased the expression of pro-inflammatory cytokine gene IL-6 but the HESM fed birds showed its effect curtailed, also, which also, favored the up-regulation of anti-inflammatory genes compared with control diet fed chickens. Post hatch supplementation of HESM appears to improve performance, modulate immunity, and increase

  2. Nutritional Supplement of Hatchery Eggshell Membrane Improves Poultry Performance and Provides Resistance against Endotoxin Stress.

    Science.gov (United States)

    Makkar, S K; Rath, N C; Packialakshmi, B; Zhou, Z Y; Huff, G R; Donoghue, A M

    2016-01-01

    Eggshells are significant part of hatchery waste which consist of calcium carbonate crust, membranes, and proteins and peptides of embryonic origins along with other entrapped contaminants including microbes. We hypothesized that using this product as a nutritional additive in poultry diet may confer better immunity to the chickens in the paradigm of mammalian milk that enhances immunity. Therefore, we investigated the effect of hatchery eggshell membranes (HESM) as a short term feed supplement on growth performance and immunity of chickens under bacterial lipopolysaccharide (LPS) challenged condition. Three studies were conducted to find the effect of HESM supplement on post hatch chickens. In the first study, the chickens were fed either a control diet or diets containing 0.5% whey protein or HESM as supplement and evaluated at 5 weeks of age using growth, hematology, clinical chemistry, plasma immunoglobulins, and corticosterone as variables. The second and third studies were done to compare the effects of LPS on control and HESM fed birds at 5 weeks of age following at 4 and 24 h of treatment where the HESM was also sterilized with ethanol to deplete bacterial factors. HESM supplement caused weight gain in 2 experiments and decreased blood corticosterone concentrations. While LPS caused a significant loss in body weight at 24 h following its administration, the HESM supplemented birds showed significantly less body weight loss compared with the control fed birds. The WBC, heterophil/lymphocyte ratio, and the levels of IgG were low in chickens fed diets with HESM supplement compared with control diet group. LPS challenge increased the expression of pro-inflammatory cytokine gene IL-6 but the HESM fed birds showed its effect curtailed, also, which also, favored the up-regulation of anti-inflammatory genes compared with control diet fed chickens. Post hatch supplementation of HESM appears to improve performance, modulate immunity, and increase resistance of

  3. Purification of LPS Binding Substances in Inflammatory Serum

    Science.gov (United States)

    1990-07-31

    fourth) utilizing skin structures as landmarks (Fig. 1). First degree burns involve the superficial epithelium only. They are dry, red, and painful to...burned hospitalized patients. 1982;22(l):11-14. 14. Spebar MJ, Pruitt BA. Candidiasis in the burned patient. J Trauma 1981;21(3):237-239. 15. Bruck HM

  4. Geographical variation and the determinants of domestic endotoxin levels in mattress dust in Europe

    NARCIS (Netherlands)

    Chen, C.M.; Thiering, E.; Doekes, G.|info:eu-repo/dai/nl/070079803; Zock, J.P.|info:eu-repo/dai/nl/095184309; Bakolis, I.; Norbäck, D.; Sunyer, J.; Villani, S.; Verlato, G.; Täubel, M.; Jarvis, D.

    2012-01-01

    Endotoxin exposures have manifold effects on human health. The geographical variation and determinants of domestic endotoxin levels in Europe have not yet been extensively described. To investigate the geographical variation and determinants of domestic endotoxin concentrations in mattress dust in

  5. Apigenin Alleviates Endotoxin-Induced Myocardial Toxicity by Modulating Inflammation, Oxidative Stress, and Autophagy

    Directory of Open Access Journals (Sweden)

    Fang Li

    2017-01-01

    Full Text Available Apigenin, a component in daily diets, demonstrates antioxidant and anti-inflammatory properties. Here, we intended to explore the mechanism of apigenin-mediated endotoxin-induced myocardial injury and its role in the interplay among inflammation, oxidative stress, and autophagy. In our lipopolysaccharide- (LPS- induced myocardial injury model, apigenin ameliorated cardiac injury (lactate dehydrogenase (LDH and creatine kinase (CK, cell death (TUNEL staining, DNA fragmentation, and PARP activity, and tissue damage (cardiac troponin I (cTnI and cardiac myosin light chain-1 (cMLC1 and improved cardiac function (ejection fraction (EF and end diastolic left ventricular inner dimension (LVID. Apigenin also alleviated endotoxin-induced myocardial injury by modulating oxidative stress (nitrotyrosine and protein carbonyl and inflammatory cytokines (TNF-α, IL-1β, MIP-1α, and MIP-2 along with their master regulator NFκB. Apigenin modulated redox homeostasis, and its anti-inflammatory role might be associated with its ability to control autophagy. Autophagy (determined by LAMP1, ATG5, and p62, its transcriptional regulator transcription factor EB (TFEB, and downstream target genes including vacuolar protein sorting-associated protein 11 (Vps11 and microtubule-associated proteins 1A/1B light chain 3B (Map1lc3 were modulated by apigenin. Thus, our study demonstrated that apigenin may lead to potential development of new target in sepsis treatment or other myocardial oxidative and/or inflammation-induced injuries.

  6. A System Dynamics Model to Predict the Human Monocyte Response to Endotoxins

    Directory of Open Access Journals (Sweden)

    Enrique Álvarez

    2017-08-01

    Full Text Available System dynamics is a powerful tool that allows modeling of complex and highly networked systems such as those found in the human immune system. We have developed a model that reproduces how the exposure of human monocytes to lipopolysaccharides (LPSs induces an inflammatory state characterized by high production of tumor necrosis factor alpha (TNFα, which is rapidly modulated to enter into a tolerant state, known as endotoxin tolerance (ET. The model contains two subsystems with a total of six states, seven flows, two auxiliary variables, and 14 parameters that interact through six differential and nine algebraic equations. The parameters were estimated and optimized to obtain a model that fits the experimental data obtained from human monocytes treated with various LPS doses. In contrast to publications on other animal models, stimulation of human monocytes with super-low-dose LPSs did not alter the response to a second LPSs challenge, neither inducing ET, nor enhancing the inflammatory response. Moreover, the model confirms the low production of TNFα and increased levels of C–C motif ligand 2 when monocytes exhibit a tolerant state similar to that of patients with sepsis. At present, the model can help us better understand the ET response and might offer new insights on sepsis diagnostics and prognosis by examining the monocyte response to endotoxins in patients with sepsis.

  7. The LPS-induced increase in circulating microparticles is not affected by vitamin C in humans.

    Science.gov (United States)

    Weisshaar, Stefan; Gouya, Ghazaleh; Nguyen, Daniel; Kapiotis, Stylianos; Wolzt, Michael

    2013-07-01

    Microparticles (MP) are considered to promote coagulation. This study aimed to characterize the time course of MP levels and the effect of high-dose vitamin C on MP formation during inflammation in an in vivo Escherichia coli endotoxin (LPS) model. Microparticle formation was studied in 14 male subjects in a cross-over trial who received either intravenous vitamin C at 320 mg/kg body weight (BW) or 480 mg/kg BW or saline solution in a random order on alternate trial days 3 h after intravenous exposure to LPS (2 ng/kg BW). Venous blood samples were taken before, 3 and 6 h after LPS. D-dimer, leucocyte count, C-reactive protein, plasma vitamin C and body temperature were assessed as inflammatory parameters. MP were detected using flow cytometric analysis and expressed in 10³ MP/mL plasma. Microparticles levels were decreased from baseline 848 units [range 431-1705] by 21% to 671 units [253-1586] at 3 h and increased by 32% to 1119 units [288-4443] at 6 h after LPS. This pattern was not influenced by administration of vitamin C, with a change from 730 units [399-1396] at baseline by an increase to 832 units [215-2168] at 3 h to 1055 units [350-4858] at 6 h. MP subpopulations followed similar dynamics. Alterations in inflammatory parameters were independent from vitamin C administration during endotoxemia. Microparticles are increased in acute systemic inflammation with inconsistent changes in MP subgroups in healthy subjects. Systemic vitamin C administration does not mitigate MP formation and D-dimer levels during acute systemic inflammation, suggesting that MP-induced coagulation activity is not affected by vitamin C. © 2013 Stichting European Society for Clinical Investigation Journal Foundation. Published by John Wiley & Sons Ltd.

  8. Inhibition of LPS and Plasmodium falciparum induced cytokine secretion by pentoxifylline and two analogues

    DEFF Research Database (Denmark)

    Jakobsen, P H; Koch, C; Bendtzen, K

    1997-01-01

    Pentoxifylline and the two analogues HWA138 and HWA448, at concentrations exceeding 60 micrograms/ml, inhibited malaria antigen or lipopolysaccharide (LPS) induced TNF-alpha and IL-1 alpha secretion, but not IL-6 secretion, from human peripheral blood mononuclear cells in vitro. HWA448 had lower...... in female CF1xBalb/c mice. The drugs were inhibitory at 0.5-1 mg per mouse when mixed with LPS, and 1 mg per mouse of the drugs was inhibitory when injected 1 h before LPS challenge. HWA448 had similar inhibitory activities in vivo compared to pentoxifylline and HWA138, possibly because of the longer serum...

  9. Recombinant immunotoxins with low endotoxins for clinical and animal studies.

    Science.gov (United States)

    Onda, Masanori

    2012-01-01

    Recombinant immunotoxin (RIT) contains the Fv portion of the antibody fused to the truncated form of toxin and are ongoing in clinical trials for cancer therapy. To obtain high yields of products, RITs are produced in Escherichia coli (E. coli). As the endotoxin came from E. coli cells and is harmful to animals, it is important to produce the RITs with low endotoxin. This section describes the protocols to produce RITs containing low level of endotoxins.

  10. Colistin Reduces LPS-Triggered Inflammation in a Human Sepsis Model In Vivo: A Randomized Controlled Trial.

    Science.gov (United States)

    Matzneller, P; Strommer, S; Drucker, C; Petroczi, K; Schörgenhofer, C; Lackner, E; Jilma, B; Zeitlinger, M

    2017-06-01

    The previously described anti-endotoxin effect of colistin has not been investigated in humans yet. We performed a randomized, double-blind, placebo-controlled crossover trial to determine the degree of colistin-driven modulation of inflammatory response in blood of lipopolysaccharide (LPS)-challenged healthy volunteers in a human endotoxemia model. After a single intravenous dose of 2.5 million IU colistin methanesulfonate, interleukin (IL)-6, IL-8, tumor necrosis factor alpha (TNF-α), and IL-1β concentrations as well as other biomarkers of inflammation such as C-reactive protein, differential leukocyte counts, and body temperature were measured up to 24 h postdose. Colistin significantly decreased the inflammatory cytokine response to LPS in blood of healthy volunteers. This effect was most evident for IL-6, IL-8, and TNF-α. This study is the first to confirm the anti-endotoxin effect of colistin in humans in vivo. Further studies might increase our knowledge on the interaction between colistin and the effectors of the immune system. © 2017 American Society for Clinical Pharmacology and Therapeutics.

  11. In vivo modulation of LPS induced leukotrienes generation and oxidative stress by sesame lignans.

    Science.gov (United States)

    Yashaswini, Puttaraju Srikantamurthy; Sadashivaiah, Bettadahalli; Ramaprasad, Talahalli Ravichandra; Singh, Sridevi Annapurna

    2017-03-01

    The role of inflammation and oxidative stress is critical during onset of metabolic disorders and this has been sufficiently established in literature. In the present study, we evaluated the effects of sesamol and sesamin, two important bioactive molecules present in sesame oil, on the generation of inflammatory and oxidative stress factors in LPS injected rats. Sesamol and sesamin lowered LPS induced expression of cPLA2 (61 and 56%), 5-LOX (44 and 51%), BLT-1(32 and 35%) and LTC4 synthase (49 and 50%), respectively, in liver homogenate. The diminished serum LTB4 (53 and 64%) and LTC4 (67 and 44%) levels in sesamol and sesamin administered groups, respectively, were found to be concurrent with the observed decrease in the expression of cPLA2 and 5-LOX. The serum levels of TNF-α (29 and 19%), MCP-1 (44 and 57%) and IL-1β (43 and 42%) were found to be reduced in sesamol and sesamin group, respectively, as given in parentheses, compared to LPS group. Sesamol and sesamin offered protection against LPS induced lipid peroxidation in both serum and liver. Sesamol, but not sesamin, significantly restored the loss of catalase and glutathione reductase activity due to LPS (P<.05). However, both sesamol and sesamin reverted SOD activities by 92 and 98%, respectively. Thus, oral supplementation of sesamol and sesamin beneficially modulated the inflammatory and oxidative stress markers, as observed in the present study, in LPS injected rats. Our report further advocates the potential use of sesamol and sesamin as an adjunct therapy wherein, inflammatory and oxidative stress is of major concern. Copyright © 2016 Elsevier Inc. All rights reserved.

  12. Endotoxin and cytokines in patients with gastrointestinal tract perforation

    Directory of Open Access Journals (Sweden)

    S. Endo

    1992-01-01

    Full Text Available Plasma levels of endotoxin and various cytokines were assessed in 70 patients with gastrointestinal tract perforation. Sepsis developed in 29 of them, and eight of these (27.6% had on admission endotoxin levels higher than 9.8 pg ml-1. The clinical outcome correlated with the level of tumour necrosis factor α (TNFα, rather than with the endotoxin level. The high interleukin 6 (IL-6 level was shown in septic patients and no correlation was observed between the IL-6 level and the clinical outcome. Plasma TNFα levels tended to change independently from endotoxin levels, suggesting that TNFα may have been locally produced in inflammatory lesions.

  13. Are Cockroaches an Important Source of Indoor Endotoxins?

    Science.gov (United States)

    Lai, Ka Man

    2017-01-18

    Endotoxins are common indoor biocontaminants. Their levels have been shown to link to many sources and factors. One of them is cockroach infestation but the role of cockroaches and contamination mechanisms are unclear. We hypothesized that not only is cockroach infestation a sign of poor hygiene, but it also contributes to indoor endotoxins via fecal contamination. In this study, different cockroach species were caught in homes. The endotoxin and allergen levels and their ratios in cockroach feces were determined. To estimate the amount of indoor endotoxins that originated from cockroaches, a new approach of using these new cockroach endotoxin and allergen ratios to compare with environmental data was employed. We found that Supella (S.) longipalpa, Periplaneta (P.) australasiae, and Blattella (B.) germanica were dominant in homes. On average, P. australasiae feces had a higher level but greater variation of endotoxins. B. germanica feces had the highest levels of allergens measured. Depending on environmental bacterial load and the type of cockroaches present, cockroach endotoxins in the environment may vary greatly. Cockroaches directly contribute to indoor endotoxins rather than just being a sign of poor hygiene. The type and extent of cockroach infestation should be taken into consideration when assessing and remediating indoor endotoxin contamination.

  14. Risks associated with endotoxins in feed additives produced by fermentation.

    Science.gov (United States)

    Wallace, R John; Gropp, Jürgen; Dierick, Noël; Costa, Lucio G; Martelli, Giovanna; Brantom, Paul G; Bampidis, Vasileios; Renshaw, Derek W; Leng, Lubomir

    2016-01-15

    Increasingly, feed additives for livestock, such as amino acids and vitamins, are being produced by Gram-negative bacteria, particularly Escherichia coli. The potential therefore exists for animals, consumers and workers to be exposed to possibly harmful amounts of endotoxin from these products. The aim of this review was to assess the extent of the risk from endotoxins in feed additives and to calculate how such risk can be assessed from the properties of the additive. Livestock are frequently exposed to a relatively high content of endotoxin in the diet: no additional hazard to livestock would be anticipated if the endotoxin concentration of the feed additive falls in the same range as feedstuffs. Consumer exposure will be unaffected by the consumption of food derived from animals receiving endotoxin-containing feed, because the small concentrations of endotoxin absorbed do not accumulate in edible tissues. In contrast, workers processing a dusty additive may be exposed to hazardous amounts of endotoxin even if the endotoxin concentration of the product is low. A calculation method is proposed to compare the potential risk to the worker, based on the dusting potential, the endotoxin concentration and technical guidance of the European Food Safety Authority, with national exposure limits.

  15. Quality-control analytical methods: endotoxins: essential testing for pyrogens in the compounding laboratory, part 3: a simplified endotoxin test method for compounded sterile preparations.

    Science.gov (United States)

    Cooper, James F

    2011-01-01

    The first two parts of the IJPC series on endotoxin testing explained the nature of pyrogenic contamination and described various Limulus amebocyte lysate methods for detecting and measuring endotoxin levels with the bacterial endotoxin test described in the United States Pharmacopeia. This third article in that series describes the endotoxin test that is simplest to permorm for pharmacists who prefer to conduct an endotoxin assa at the time of compounding in the pharmacy setting.

  16. Endotoxin/lipopolysaccharide activates NF-kappa B and enhances tumor cell adhesion and invasion through a beta 1 integrin-dependent mechanism.

    LENUS (Irish Health Repository)

    Wang, Jiang Huai

    2012-02-03

    Beta(1) integrins play a crucial role in supporting tumor cell attachment to and invasion into the extracellular matrix. Endotoxin\\/LPS introduced by surgery has been shown to enhance tumor metastasis in a murine model. Here we show the direct effect of LPS on tumor cell adhesion and invasion in extracellular matrix proteins through a beta(1) integrin-dependent pathway. The human colorectal tumor cell lines SW480 and SW620 constitutively expressed high levels of the beta(1) subunit, whereas various low levels of alpha(1), alpha(2), alpha(4), and alpha(6) expression were detected. SW480 and SW620 did not express membrane-bound CD14; however, LPS in the presence of soluble CD14 (sCD14) significantly up-regulated beta(1) integrin expression; enhanced tumor cell attachment to fibronectin, collagen I, and laminin; and strongly promoted tumor cell invasion through the Matrigel. Anti-beta(1) blocking mAbs (4B4 and 6S6) abrogated LPS- plus sCD14-induced tumor cell adhesion and invasion. Furthermore, LPS, when combined with sCD14, resulted in NF-kappaB activation in both SW480 and SW620 cells. Inhibition of the NF-kappaB pathway significantly attenuated LPS-induced up-regulation of beta(1) integrin expression and prevented tumor cell adhesion and invasion. These results provide direct evidence that although SW480 and SW620 cells do not express membrane-bound CD14, LPS in the presence of sCD14 can activate NF-kappaB, up-regulate beta(1) integrin expression, and subsequently promote tumor cell adhesion and invasion. Moreover, LPS-induced tumor cell attachment to and invasion through extracellular matrix proteins is beta(1) subunit-dependent.

  17. Organ-specific and differential requirement of TYK2 and IFNAR1 for LPS-induced iNOS expression in vivo.

    Science.gov (United States)

    Painz, Ronald; Walter, Ingrid; Kolbe, Thomas; Rigler, Doris; Vogl, Claus; Steinborn, Ralf; Rülicke, Thomas; Helmreich, Magdalena; Karaghiosoff, Marina; Müller, Mathias

    2007-01-01

    Lipopolysaccharide (LPS) is an integral structural component of the outer membrane of Gram-negative bacteria and the principal active agent in the pathogenesis of endotoxin shock. LPS is a potent inducer of a variety of cytokines and inflammatory agents that lead to a profound alteration of gene expression patterns in cells and organs. The gene coding for the inducible nitric oxide synthase (iNOS) is highly responsive to LPS in vitro and in vivo and accounts for the production of nitric oxide (NO). The Janus kinase (JAK) family member tyrosine kinase 2 (TYK2) is a constituent of the interferon (IFN) type I response pathway and an important effector in the progression of endotoxin shock. Macrophages deficient for IFNalphabeta receptor chain 1 (IFNAR1) or TYK2 were shown to have an impaired LPS-induced iNOS expression. Here we determined the contribution of IFNAR1 and TYK2 to iNOS expression in vivo in a lethal LPS challenge model. TYK2 and IFNAR1 were found to be crucial for the LPS-induced iNOS mRNA and protein expression in spleen and lung that could be attributed to the Mac3-positive population. In liver LPS-induced iNOS mRNA expression was only partially impaired in TYK2-deficient mice and was unimpaired in IFNAR1-deficient mice, indicating organ specificity. TYK2(-/-) and IFNAR1(-/-) mice also differ with respect to IFNgamma production upon LPS challenge in that TYK2(-/-) mice show a defect while IFNAR1(-/-) mice do not. Our data suggest that iNOS is induced through IFNAR1 and TYK2 in Mac3-positive cells which are the main source of iNOS in spleen and lung. The LPS-induced iNOS expression in liver is independent of IFNAR1 and partially dependent on TYK2, which is most likely due to the lack of IFNgamma production in the absence of TYK2.

  18. PATTERNS OF CYTOKINES, PLASMA ENDOTOXIN, PLASMINOGEN-ACTIVATOR INHIBITOR, AND ACUTE-PHASE PROTEINS DURING THE TREATMENT OF SEVERE SEPSIS IN HUMANS

    NARCIS (Netherlands)

    DOFFERHOFF, ASM; BOM, VJJ; DEVRIESHOSPERS, HG; VANINGEN, J; VANDERMEER, J; HAZENBERG, BPC; MULDER, POM; WEITS, J

    Objective: To study the patterns of plasma concentrations of endotoxin, tumor necrosis factor-alpha (TNF), interleukin-6 (IL-6), plasminogen activator inhibitor-1, C-reactive protein, and serum amyloid A during the treatment of human sepsis. Design: A prospective case series study. Setting: ICU of

  19. Combined Insulin Deficiency and Endotoxin Exposure Stimulate Lipid Mobilization and Alter Adipose Tissue Signaling in an Experimental Model of Ketoacidosis in Subjects with Type 1 Diabetes: A Randomized, Controlled, Crossover Trial

    DEFF Research Database (Denmark)

    Svart, Mads; Opstrup, Ulla Kampmann; Voss, Thomas

    2016-01-01

    and endotoxin administration(KET).During KET, serum TNF-α, cortisol, glucagon, and growth hormone levels increased; free fatty acids and 3-hydroxybutyrate concentrations and the rate of lipolysis rose markedly. Serum bicarbonate and pH decreased. Adipose tissue mRNA contents of comparative gene identification...

  20. Expression Profile of Cationic Amino Acid Transporters in Rats with Endotoxin-Induced Uveitis

    Directory of Open Access Journals (Sweden)

    Yung-Ray Hsu

    2016-01-01

    Full Text Available Purpose. The transcellular arginine transportation via cationic amino acid transporter (CAT is the rate-limiting step in nitric oxide (NO synthesis, which is crucial in intraocular inflammation. In this study, CAT isoforms and inducible nitric oxide synthase (iNOS expression was investigated in endotoxin-induced uveitis (EIU. Methods. EIU was induced in Lewis rats by lipopolysaccharide (LPS injection. In the treatment group, the rats were injected intraperitoneally with the proteasome inhibitor bortezomib before EIU induction. After 24 hours, leukocyte quantification, NO measurement of the aqueous humor, and histopathological examination were evaluated. The expression of CAT isoforms and iNOS was determined by reverse transcription-polymerase chain reaction, western blotting, and immunofluorescence staining. Nuclear factor-kappa B (NF-κB binding activity was evaluated by electrophoretic mobility shift assay. The mouse macrophage cell line RAW 264.7 was used to validate the in vivo findings. Results. LPS significantly stimulated iNOS, CAT-2A, and CAT-2B mRNA and protein expression but did not affect CAT-1 in EIU rats and RAW 264.7 cells. Bortezomib attenuated inflammation and inhibited iNOS, CAT-2A, and CAT-2B expression through NF-κB inhibition. Conclusions. CAT-2 and iNOS, but not CAT-1, are specifically involved in EIU. NF-κB is essential in the induction of CAT-2 and iNOS in EIU.

  1. The Aldosterone-Mineralocorticoid Receptor Pathway Exerts Anti-Inflammatory Effects in Endotoxin-Induced Uveitis

    Science.gov (United States)

    Ly, André; Leroux les Jardins, Guillaume; Goldenberg, Brigitte; Naud, Marie-Christine; Jonet, Laurent; Besson-Lescure, Bernadette; Jaisser, Frederic; Farman, Nicolette; De Kozak, Yvonne; Behar-Cohen, Francine

    2012-01-01

    We have previously shown that the eye is a mineralocorticoid-sensitive organ and we now question the role of mineralocorticoid receptor (MR) in ocular inflammation. The endotoxin-induced uveitis (EIU), a rat model of human intraocular inflammation, was induced by systemic administration of lipopolysaccharide (LPS). Evaluations were made 6 and 24 hours after intraocular injection of aldosterone (simultaneous to LPS injection). Three hours after onset of EIU, the MR and the glucocorticoid metabolizing enzyme 11-beta hydroxysteroid dehydrogenase type 2 (11β-HSD2) expression were down-regulated in iris/ciliary body and the corticosterone concentration was increased in aqueous humor, altering the normal MR/glucocorticoid receptor (GR) balance. At 24 hours, the GR expression was also decreased. In EIU, aldosterone reduced the intensity of clinical inflammation in a dose-dependent manner. The clinical benefit of aldosterone was abrogated in the presence of the MR antagonist (RU26752) and only partially with the GR antagonist (RU38486). Aldosterone reduced the release of inflammatory mediators (6 and 24 hours: TNF-α, IFN-γ, MIP-1α) in aqueous humor and the number of activated microglia/macrophages. Aldosterone partly prevented the uveitis-induced MR down-regulation. These results suggest that MR expression and activation in iris/ciliary body could protect the ocular structures against damages induced by EIU. PMID:23152847

  2. Amino acid supplementation is anabolic during the acute phase of endotoxin-induced inflammation: A human randomized crossover trial.

    Science.gov (United States)

    Rittig, N; Bach, E; Thomsen, H H; Johannsen, M; Jørgensen, J O; Richelsen, B; Jessen, N; Møller, N

    2016-04-01

    Inflammation is catabolic and causes muscle loss. It is unknown if amino acid supplementation reverses these effects during the acute phase of inflammation. The aim was to test whether amino acid supplementation counteracts endotoxin-induced catabolism. Eight young, healthy, lean males were investigated three times in randomized order: (i) normal conditions (Placebo), (ii) endotoxemia (LPS), and (iii) endotoxemia with amino acid supplementation (LPS + A). Protein kinetics were determined using phenylalanine, tyrosine, and urea tracers. Each study day consisted of a four-hour non-insulin stimulated period and a two-hour hyperinsulinemic euglycemic clamp period. Muscle biopsies were collected once each period. Endotoxin administration created a significant inflammatory response (cytokines, hormones, and vital parameters) without significant differences between LPS and LPS + A. Whole body protein breakdown was elevated during LPS compared with Placebo and LPS + A (p translation factor 4E-binding protein 1 (4EBP1) phosphorylation (p = 0.007) without activating AMPK or affecting insulin signaling through Akt. During insulin stimulation net muscle phenylalanine release and protein degradation were further reduced. Amino acid supplementation in the acute phase of inflammation reduces whole body and muscle protein loss, and this effect is associated with activation of mTOR and downstream signaling to protein synthesis through mTORC1, suggesting a therapeutic role for intravenous amino acids in inflammatory states. The Central Denmark Region Ethics Commitee (1-10-71-410-12) www.clinicaltrials.gov (identification number NCT01705782). Copyright © 2015 Elsevier Ltd and European Society for Clinical Nutrition and Metabolism. All rights reserved.

  3. A signal-on electrochemical aptasensor for ultrasensitive detection of endotoxin using three-way DNA junction-aided enzymatic recycling and graphene nanohybrid for amplification

    Science.gov (United States)

    Bai, Lijuan; Chai, Yaqin; Pu, Xiaoyun; Yuan, Ruo

    2014-02-01

    Endotoxin, also known as lipopolysaccharide (LPS), is able to induce a strong immune response on its internalization into mammalian cells. To date, aptamer-based biosensors for LPS detection have been rarely reported. This work describes a new signal-on electrochemical aptasensor for the ultrasensitive detection of LPS by combining the three-way DNA hybridization process and nanotechnology-based amplification. With the help of DNA1 (associated with the concentration of target LPS), the capture probe hybridizes with DNA1 and the assistant probe to open its hairpin structure and form a ternary ``Y'' junction structure. The DNA1 can be released from the structure in the presence of nicking endonuclease to initiate the next hybridization process. Then a great deal of cleaved capture probe produced in the cyclic process can bind with DNA2-nanocomposite, which contains the electroactive toluidine blue (Tb) with the amplification materials graphene (Gra) and gold nanoparticles (AuNPs). Thus, an enhanced electrochemical signal can be easily read out. With the cascade signal amplification, this newly designed protocol provides an ultrasensitive electrochemical detection of LPS down to the femtogram level (8.7 fg mL-1) with a linear range of 6 orders of magnitude (from 10 fg mL-1 to 50 ng mL-1). Moreover, the high sensitivity and specificity make this method versatile for the detection of other biomolecules by changing the corresponding sequences of the capture probe and the assistant probe.

  4. Efficacy of rifaximin on circulating endotoxins and cytokines in patients with nonalcoholic fatty liver disease.

    Science.gov (United States)

    Gangarapu, Venkatanarayana; Ince, Ali Tüzün; Baysal, Birol; Kayar, Yusuf; Kılıç, Ulkan; Gök, Özlem; Uysal, Ömer; Şenturk, Hakan

    2015-07-01

    Recent studies have suggested that endotoxin-induced cytokines play an important role in nonalcoholic fatty liver disease (NAFLD). Rifaximin is a nonabsorbable antibiotic that might act on Gram-negative bacteria, thereby inhibiting endotoxin proinflammatory cytokine production in patients with NAFLD. Our aim was to investigate the efficacy of rifaximin on NAFLD. Forty-two patients with biopsy-proven NAFLD [15 steatosis, 27 nonalcoholic steatohepatitis (NASH)] were included in this prospective, open-label, observational cohort study. BMI and serum aspartate aminotransferase, alanine aminotransferase (ALT), gamma glutamyl transferase, lipid profile, ferritin, C-reactive protein, glucose, insulin, homeostatic model assessment as well as endotoxin, serum Toll-like receptor 4 (TlR4), interleukin-1α (IL-1α), IL-6, IL-10, IL-12, and tumor necrosis factor-α (TNF-α) levels were measured before and after a 28-day administration of rifaximin (1200 mg/daily). Results were analyzed using nonparametric Wilcoxon signed-rank tests. A mild reduction in the mean BMI (32.3 ± 6.9 vs. 31.9 ± 6.8, P = 0.02) and a significant reduction in the endotoxin (0.9 ± 0.34 vs. 0.8 ± 0.13, P = 0.03) and IL-10 (4.08 ± 0.9 vs. 3.73 ± 0.7, P = 0.006) levels in the NASH group were noted. A significant reduction was observed in serum aspartate aminotransferase (50.4 ± 39 vs. 33 ± 14, P = 0.01), ALT (72 ± 48 vs. 45.2 ± 26.3, P = 0.0001), gamma glutamyl transferase (52 ± 33 vs. 41.2 ± 21.1, P = 0.02), LDL (137 ± 34 vs. 127 ± 27.5, P = 0.03), and ferritin (142 ± 214 vs. 89.3 ± 123, P = 0.0001) in the NASH group, but only in ALT (50.4 ± 26 vs. 35.5 ± 23.25, P = 0.01), and ferritin (73.6 ± 83 vs. 55 ± 76, P = 0.004) levels decreased significantly in the steatosis group. Treatment with rifaximin did not exert a significant effect on serum levels of TLR-4, IL-1, IL-6, IL-12, or TNF-α in either group. In NAFLD and especially in NASH, short-term administration of rifaximin appears to be

  5. Detection of Bacteroides fragilis endotoxin in amniotic fluid by counterimmunoelectrophoresis

    NARCIS (Netherlands)

    I. Beckmann (Ilse); K. de Graaff (K.); F. Meisel-Mikolajczyk; H.C.S. Wallenburg (Henk)

    1994-01-01

    textabstractThe ability of counter immunoelectrophoresis (CIE) to detectBacteroides fragilis endotoxin in amniotic fluid in small concentrations was evaluated. A method was developed which, in combination with ultrafiltration, permits detection ofB. fragilis endotoxin in amniotic fluid in a

  6. The redistribution of granulocytes following E. coli endotoxin induced sepsis

    DEFF Research Database (Denmark)

    Toft, P; Lillevang, S T; Tønnesen, Else Kirstine

    1994-01-01

    Infusion of endotoxin elicits granulocytopenia followed by increased numbers of granulocytes in peripheral blood. The purpose of this study was to investigate the redistribution and sequestration of granulocytes in the tissues following E. coli endotoxin induced sepsis. From 16 rabbits granulocytes...

  7. Evaluation of endotoxin retention by adsorptive-based filtration media.

    Science.gov (United States)

    O'Brien, Thomas P; Conway, Robert; Chen, Hsiao-Lin; Buckland, Kim

    2007-01-01

    Control of endotoxin contamination is an important issue in pharmaceutical and bioprocess manufacturing. Endotoxins can contaminate process intermediates used in pharmaceutical formulations, aqueous- and non-aqueous-based CIP fluids used in equipment and vial cleaning, and process fluids such as buffers used for chromatographic elution, diafiltration, and suspension of therapeutic protein-based drugs. A study was undertaken to evaluate the effectiveness of adsorptive-based depth and membrane filtration media in removing suspended endotoxin. The following variables were examined in order to determine their effects on endotoxin reduction: absorptive media type, residence time (flux), challenge solution pH, and interferences in endotoxin reduction as the result of challenge solution composition-water for injection, process buffer, and the presence of protein. The endotoxin removal capacities of the various media studied were also determined. The results of the study demonstrated differences in the effect on endotoxin removal of the variables evaluated. In addition, the results provide a strategy for conducting studies to select and validate an appropriate adsorptive filter media for control of endotoxin contamination.

  8. Eukaryotic elongation factor 2 controls TNF-α translation in LPS-induced hepatitis

    Science.gov (United States)

    González-Terán, Bárbara; Cortés, José R.; Manieri, Elisa; Matesanz, Nuria; Verdugo, ρngeles; Rodríguez, María E.; González-Rodríguez, ρgueda; Valverde, ρngela; Martín, Pilar; Davis, Roger J.; Sabio, Guadalupe

    2012-01-01

    Bacterial LPS (endotoxin) has been implicated in the pathogenesis of acute liver disease through its induction of the proinflammatory cytokine TNF-α. TNF-α is a key determinant of the outcome in a well-established mouse model of acute liver failure during septic shock. One possible mechanism for regulating TNF-α expression is through the control of protein elongation during translation, which would allow rapid cell adaptation to physiological changes. However, the regulation of translational elongation is poorly understood. We found that expression of p38γ/δ MAPK proteins is required for the elongation of nascent TNF-α protein in macrophages. The MKK3/6-p38γ/δ pathway mediated an inhibitory phosphorylation of eukaryotic elongation factor 2 (eEF2) kinase, which in turn promoted eEF2 activation (dephosphorylation) and subsequent TNF-α elongation. These results identify a new signaling pathway that regulates TNF-α production in LPS-induced liver damage and suggest potential cell-specific therapeutic targets for liver diseases in which TNF-α production is involved. PMID:23202732

  9. Removal of viable bacteria and endotoxins by Electro Deionization (EDI).

    Science.gov (United States)

    Harada, Norimitsu; Otomo, Teruo; Watabe, Tomoichi; Ase, Tomonobu; Takemura, Takuto; Sato, Toshio

    2011-09-01

    Viable bacteria and endotoxins in water sometimes cause problems for human health. Endotoxins are major components of the outer cell wall of gram-negative bacteria (lipopolysaccharides). In medical procedures, especially haemodialysis (HD) and related therapies (haemodiafiltration (HDF), haemofiltration (HF)), endotoxins in the water for haemodialysis can permeate through the haemodialysis membrane and cause microinflammation or various haemodialysis-related illnesses. To decrease such a biological risk, RO and UF membranes are generally used. Also, hot water disinfection or the chemical disinfection is regularly executed to kill bacteria which produce endotoxins. However, simple treatment methods and equipment may be able to decrease the biological risk more efficiently. In our experiments, we confirmed that viable bacteria and endotoxins were removed by Electro Deionization (EDI) technology and also clarified the desorption mechanisms.

  10. Endotoxins in tobacco smoke: shifting tobacco industry positions.

    Science.gov (United States)

    Barnes, Richard L; Glantz, Stanton A

    2007-10-01

    In the 1980s, the tobacco industry started a campaign to divert attention away from secondhand tobacco smoke (SHS) as a major source of indoor air pollution in workplaces by highlighting the roles of other indoor air pollutants. The industry, working through "third parties," highlighted endotoxins, naturally occurring substances that cause numerous inflammatory reactions in humans, as an alternative explanation to SHS as causing indoor air problems. In 1995, Hasday and colleagues were the first to present findings that cigarette smoke contains significant quantities of endotoxins. This discovery surprised tobacco industry scientists. The 1999 publication of the full Hasday et al. findings received only limited media attention but got the full attention of Philip Morris scientists concerned about a new public health issue and a new basis for regulation of workplace smoking by the U.S. Occupational Safety and Health Administration, which already regulated workplace endotoxin exposures from other sources. Philip Morris undertook an internal endotoxin research project to test the Hasday et al. findings and to determine if endotoxin-free cigarettes were possible. Although experiments were conducted to remove endotoxin from the tobacco, there is no evidence that they were successful. Following confirmation of SHS as an important source of endotoxins, the scientist promoting endotoxins as an important indoor air pollutant for the tobacco industry softened his position on the role of endotoxins as indoor pollutants. The presence of endotoxins in SHS provides an additional mechanism for the adverse effects of SHS that should be researched further, and the risk of exposure should be assessed.

  11. Endotoxin and CD14 in the progression of biliary atresia

    Directory of Open Access Journals (Sweden)

    Chen Ching-Mei

    2010-12-01

    Full Text Available Abstract Background Biliary atresia (BA is a typical cholestatic neonatal disease, characterized by obliteration of intra- and/or extra-hepatic bile ducts. However, the mechanisms contributing to the pathogenesis of BA remain uncertain. Because of decreased bile flow, infectious complications and damaging endotoxemia occur frequently in patients with BA. The aim of this study was to investigate endotoxin levels in patients with BA and the relation of these levels with the expression of the endotoxin receptor, CD14. Methods The plasma levels of endotoxin and soluble CD14 were measured with a pyrochrome Limulus amebocyte lysate assay and enzyme-linked immunosorbent assay in patients with early-stage BA when they received the Kasai procedure (KP, in patients who were jaundice-free post-KP and followed-up at the outpatient department, in patients with late-stage BA when they received liver transplantation, and in patients with choledochal cysts. The correlation of CD14 expression with endotoxin levels in rats following common bile duct ligation was investigated. Results The results demonstrated a significantly higher hepatic CD14 mRNA and soluble CD14 plasma levels in patients with early-stage BA relative to those with late-stage BA. However, plasma endotoxin levels were significantly higher in both the early and late stages of BA relative to controls. In rat model, the results demonstrated that both endotoxin and CD14 levels were significantly increased in liver tissues of rats following bile duct ligation. Conclusions The significant increase in plasma endotoxin and soluble CD14 levels during BA implies a possible involvement of endotoxin stimulated CD14 production by hepatocytes in the early stage of BA for removal of endotoxin; whereas, endotoxin signaling likely induced liver injury and impaired soluble CD14 synthesis in the late stages of BA.

  12. Effect of enteral versus parenteral nutrition on LPS-induced sepsis in a rat model.

    Science.gov (United States)

    Hagiwara, Satoshi; Iwasaka, Hideo; Matsumoto, Shigekiyo; Noguchi, Takayuki

    2008-04-01

    The purpose of the present study was to determine whether total enteral nutrition (TEN) or total parenteral nutrition (TPN) differ in their modulation of ghrelin production and cardiac dysfunction induced by lipopolysaccharide (LPS). Vascular catheters or gastrostomy tubes were surgically placed into rats who received isocaloric parenteral or enteral nutrition postoperatively. After 7 d, the rats were injected intravenously with LPS (2.5 mg/kg). Serum ghrelin levels were determined by enzyme-linked immunosorbent assay and myocardiac function was assessed via the Langendorff isolated heart technique. Before and after the administration of LPS, TEN was found to be more effective at increasing the plasma ghrelin levels than TPN. After LPS administration, left-ventricular developed pressure decreased in animals receiving TPN when compared with animals receiving TEN. Animals receiving TPN also had significant reductions in their maximal rates of increase (+dp/dt max) and decrease (-dp/dt max) in left ventricular pressure when compared with animals receiving TEN (unpaired t-test, P maintaining cardiac function during LPS-induced septic shock.

  13. Ilexgenin A, a novel pentacyclic triterpenoid extracted from Aquifoliaceae shows reduction of LPS-induced peritonitis in mice.

    Science.gov (United States)

    Sun, Weidong; Liu, Chang; Zhang, Yaqi; Qiu, Xia; Zhang, Li; Zhao, Hongxia; Rong, Yi; Sun, Yun

    2017-02-15

    Ilexgenin A (IA) is a novel pentacyclic triterpenoid, which extracted from leaves of Ilex hainanensis Merr. In the present study, we aim to explore anti-inflammatory activity of IA on LPS-induced peritonitis and its underlying molecular mechanism. The results determined that IA was capable of suppressing peritonitis in mice induced by intraperitoneal (i.p.) injection of lipopolysaccaride (LPS). Furthermore, the results showed that IA dramatically inhibited levels of inflammatory cells infiltration in peritoneal cavity and serum in LPS-induced mice peritonitis model. Besides, IA could dramatically inhibit levels of inflammatory cytokines (IL-1β, IL-6 and TNF-α) in peritoneal cavity in LPS-induced mice peritonitis model. In vitro study, the results showed that IA inhibited production of IL-1β, IL-6 and TNF-α at transcriptional and translational levels in RAW 264.7 cells induced by LPS. Furthermore, IA could suppress the LPS-induced activation of Akt and downstream degradation and phosphorylation of kappa B-α (IκB-α). Moreover, IA could significantly inhibit ERK 1/2 phosphorylation in RAW 264.7 cells induced by LPS. These results were concurrent with molecular docking which revealed ERK1/2 inhibition. These results demonstrated that IA might as an anti-inflammatory agent candidate for inflammatory disease therapy. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Influence of the ward colon tumor on the innate and endotoxin-induced inflammatory response of the rat.

    Science.gov (United States)

    Grossie, V B

    2001-01-01

    Cachexia, a depressed immune function, and an increased infection rate are recurring problems for cancer patients; the response of the host to an infection may also be increased. We have reported that the transplantable Ward colon tumor (WCT) enhanced the lethality of endotoxin (lipopolysaccharide [LPS]) to the host. The mechanism of this increased LPS lethality and the effect of the presence of the WCT on the host inflammatory response, however, have not been reported. The effect of a transplantable WCT on the innate and endotoxin-induced inflammatory response of rats was, therefore, investigated. The innate inflammatory response was investigated in two ways. First, the formation of a granuloma around a sterile string implanted subcutaneously for 6 days was determined. Second, the effect of tumor presence on the trafficking of leukocytes to a sponge implanted subcutaneously for 2 or 6 days was determined. The tumor decreased the foreign body granuloma formation around a sterile string. The presence of the WCT also significantly blunted the increase of lymphocytes that migrated to the sponge area at 6 days. There was no significant effect of the WCT, however, on the migration of neutrophils, monocytes, or eosinophils to the implanted sponge. The leukocyte distribution of the peripheral blood was not affected by the presence of the WCT or implantation of a sterile string or sponge. To determine the effect of the presence of the WCT on the response of the host to an infection, rats were given LPS (5 mg/kg, i.p.) and the alanine amino transferase (ALT) of plasma and inducible isoforms of nitric oxide synthase (NOS2) protein content of liver, spleen, and terminal ileum was determined. The LPS challenge resulted in an increase in plasma ALT concentrations and NOS2 protein content of liver and spleen, but not the terminal ileum of WCT rats. This elevation in WCT rats confirms that the enhanced LPS-related toxicity in WCT rats was related to an increased liver toxicity

  15. Vitamin D₂ from UVB light exposed mushrooms modulates immune response to LPS in rats.

    Science.gov (United States)

    Babu, Uma S; Balan, Kannan V; Garthoff, Larry H; Calvo, Mona S

    2014-02-01

    Poor vitamin D (vitD) status is linked to increased risk of infectious diseases, thus there is need for vitD-rich foods. UVB-exposed mushrooms synthesize vitD₂ but knowledge of bioavailability and function in immune response is lacking. One hundred rats were fed one of five diets--control, 20 IU vitD₃/day; no vitD₃/day; 5% unexposed mushroom, 2.4 IU vitD₂/day; 2.5% UVB mushroom, 300 IU vitD₂/day; and 5% UVB mushroom, 600 IU vitD₂/day--for 10 wk and challenged with either saline or the endotoxin LPS. Blood and tissues were collected at 3 h postchallenge. Plasma 25-hydroxyvitamin D (25OHD) levels from UVB-exposed mushroom fed rats were significantly elevated and associated with higher natural killer cell activity and reduced plasma inflammatory response to LPS compared to control diet fed rats. Microarray evaluation of rat spleens for changes in inflammatory gene expression showed significant upregulation of proinflammatory genes after LPS compared to saline controls in all groups. However, compared to control rats, upregulation of the proinflammatory genes was markedly reduced in the groups fed vitD₂-enriched mushrooms. Rats fed UVB-exposed mushrooms had significantly higher plasma total 25OHD levels that were associated with increased innate immune response and anti-inflammatory effects. Published 2013 by Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim. This article is a U.S. Government work and is in the public domain in the USA.

  16. LPS exposure increases maternal corticosterone levels, causes placental injury and increases IL-1Β levels in adult rat offspring: relevance to autism.

    Directory of Open Access Journals (Sweden)

    Thiago B Kirsten

    Full Text Available Maternal immune activation can induce neuropsychiatric disorders, such as autism and schizophrenia. Previous investigations by our group have shown that prenatal treatment of rats on gestation day 9.5 with lipopolysaccharide (LPS; 100 μg/kg, intraperitoneally, which mimics infections by gram-negative bacteria, induced autism-like behavior in male rats, including impaired communication and socialization and induced repetitive/restricted behavior. However, the behavior of female rats was unchanged. Little is known about how LPS-induced changes in the pregnant dam subsequently affect the developing fetus and the fetal immune system. The present study evaluated the hypothalamic-pituitary-adrenal (HPA axis activity, the placental tissue and the reproductive performance of pregnant Wistar rats exposed to LPS. In the adult offspring, we evaluated the HPA axis and pro-inflammatory cytokine levels with or without a LPS challenge. LPS exposure increased maternal serum corticosterone levels, injured placental tissue and led to higher post-implantation loss, resulting in fewer live fetuses. The HPA axis was not affected in adult offspring. However, prenatal LPS exposure increased IL-1β serum levels, revealing that prenatal LPS exposure modified the immune response to a LPS challenge in adulthood. Increased IL-1β levels have been reported in several autistic patients. Together with our previous studies, our model induced autistic-like behavioral and immune disturbances in childhood and adulthood, indicating that it is a robust rat model of autism.

  17. Intestinal radiation syndrome: sepsis and endotoxin

    Energy Technology Data Exchange (ETDEWEB)

    Geraci, J.P.; Jackson, K.L.; Mariano, M.S.

    1985-03-01

    Rats were whole-body irradiated with 8-MeV cyclotron-produced neutrons and /sup 137/Cs ..gamma.. rays to study the role of enteric bacteria and endotoxin in the intestinal radiation syndrome. Decrease in intestinal weight was used as an index of radiation-induced breakdown of the mucosa. Neutron and ..gamma..-ray doses that were sublethal for intestinal death resulted in a dose-dependent decrease in intestinal weight, reaching minimal values 2 to 3 days after exposure, followed by recovery within 5 days after irradiation. Neutron and photon doses that caused intestinal death resulted in greater mucosal breakdown with little or no evidence of mucosal recovery. The presence of fluid in the intestine and diarrhea, but not bacteremia or endotoxemia, were related to mucosal breakdown and recovery. Neither sepsis nor endotoxin could be detected in liver samples taken at autopsy from animals which died a short time earlier from intestinal injury. These results suggest that overt sepsis and endotoxemia do not play a significant role in the intestinal radiation syndrome.

  18. Effect of Escherichia coli endotoxin on Archachatina marginata haemolymph coagulation system

    OpenAIRE

    Musa Salawu; Oyelola Oloyede; Adenike Oladiji; Nasiru Muhammad; Musa Yakubu

    2010-01-01

    The effect of _E. coli_ endotoxin on the heamolymph coagulation response of _Archachatina marginata_ was studied. Heamocyte Lysate(HL), Haemocyte Lysate Supernatant(HLS) and Haemocyte Lysate Debris (HLD) were exposed to _Escherichia coli_ endotoxin. Controls were prepared with endotoxin-free water(<0.025 EU/ml). The differential protein coagulation was estimated in each mixture. Fractions of the haemolymph exposed to endotoxin produced higher protein coagulates than endotoxin-free fracti...

  19. Environmental Exposure to Endotoxin and Decreased Risk of Childhood Atopy

    Directory of Open Access Journals (Sweden)

    Tatsuo Sakamoto

    2005-01-01

    Full Text Available The hygiene hypothesis implies, in immunological terms, that microbial stimulation in early life skews the immune system towards the development of a T helper type 1 (Th1-type lymphocyte population, and away from a T helper type 2 (Th2-type development that is associated with atopy. The hygiene hypothesis originally theorized that increased infections protected against atopy. This theory has evolved since harmful infections may not be as critical as exposure to microbial burden because exposure to microbes can occur in the absence of infections. Despite substantial public hygiene measures in modern metropolitan communities, an atopy-protective effect of endotoxin in these metropolitan environments still occurs. This article is a review of clinical and experimental studies concerning the protective effect of endotoxin exposure on the susceptibility to atopic responses. The discussion includes : (1 factors influencing household endotoxin levels ; (2 recent developments in the potential use of endotoxin for the prevention of atopy and asthma ; (3 association of endotoxin and pets with atopic sensitization and diseases ; (4 animal studies on the protective effects of timing of endotoxin exposure on atopy and asthma and (5 animal studies on the protective effects of endotoxin dose on atopy and asthma.

  20. Endotoxin-Mediated Downregulation of Hepatic Drug Transporters in HIV-1 Transgenic Rats.

    Science.gov (United States)

    Ghoneim, Ragia H; Piquette-Miller, Micheline

    2016-05-01

    Altered expression of drug transporters and metabolic enzymes is known to occur in infection-induced inflammation. We hypothesize that in human immunodeficiency virus (HIV)-infected individuals, further alteration could occur as a result of augmented inflammation. The HIV-1 transgenic (Tg) rat is used to simulate HIV pathologies associated with the presence of HIV viral proteins. Therefore, the objective of this study was to examine the effect of endotoxin administration on the gene expression of drug transporters in the liver of HIV-Tg rats. Male and female HIV-Tg and wild-type (WT) littermates were injected with 5 mg/kg endotoxin or saline (n= 7-9/group). Eighteen hours later, rats were euthanized and tissues were collected. Quantitative real-time polymerase chain reaction and Western blot analysis were used to measure hepatic gene and protein expression, respectively, and enzyme-linked immunosorbent assay was used to measure serum cytokine levels. Although an augmented inflammatory response was seen in HIV-Tg rats, similar endotoxin- mediated downregulation of Abcb1a, Abcc2, Abcg2, Abcb11, Slco1a1, Slco1a2, Slco1b2, Slc10a1, Slc22a1, Cyp3a2, and Cyp3a9 gene expression was seen in the HIV-Tg and WT groups. A significantly greater endotoxin- mediated downregulation of Ent1/Slc29a1 was seen in female HIV-Tg rats. Basal expression of inflammatory mediators was not altered in the HIV-Tg rat; likewise, the basal expression of most transporters was not significantly different between HIV-Tg and WT rats. Our findings suggest that hepatobiliary clearances of endogenous and exogenous substrates are altered in the HIV-Tg rat after endotoxin exposure. This is of particular importance because HIV-infected individuals frequently present with bacterial or viral infections, which are a potential source for drug-disease interactions. Copyright © 2016 by The American Society for Pharmacology and Experimental Therapeutics.

  1. Systemic endotoxin increases L-tryptophan, 5-hydroxyindoleacetic acid, 3-hydroxykynurenine and quinolinic acid content of mouse cerebral cortex.

    Science.gov (United States)

    Heyes, M P; Quearry, B J; Markey, S P

    1989-07-03

    Systemic infections and injection of endotoxin are known to increase L-tryptophan release from skeletal muscle and increase systemic L-tryptophan catabolism through the kynurenine pathway. To investigate the effects of systemically administered endotoxin on brain L-tryptophan metabolites. C57BL6/6NCR mice were given an intraperitoneal injection of 10 micrograms of lipopolysaccharide from Salmonella abortus equii and samples of serum and cerebral cortex collected. After 9 h, serum L-tryptophan concentration was decreased by 51%. At 9 h and 24 h, increases in L-tryptophan metabolites in cerebral cortex were: L-tryptophan, 42% and 39%; 5-hydroxyindoleacetic acid, 38% and 67%; 3-hydroxykynurenine, 235% and 381%; and quinolinic acid, 76% and 306%. Cortical quinolinic acid concentration was still elevated at 48 h (88%) and 72 h (79%) after lipopolysaccharide. No significant changes in cortical serotonin concentrations were found at the time points examined. When L-tryptophan (0.37 mmol/kg) was administered systemically to either normal or lipopolysaccharide-treated mice, increases in cortical L-tryptophan, serotonin, 5-hydroxyindoleacetic acid and 3-hydroxykynurenine concentrations were largest in mice treated with both lipopolysaccharide and L-tryptophan. These results suggest that disturbances in L-tryptophan metabolism that follow systemic endotoxin administration extend to the central nervous system. The consequences of these changes in L-tryptophan metabolites remain to be determined.

  2. Determination of endotoxins in caries: association with pulpal pain.

    Science.gov (United States)

    Khabbaz, M G; Anastasiadis, P L; Sykaras, S N

    2000-03-01

    The aims of this study were: (i) to determine the presence or absence of endotoxins in the superficial and deep layers of carious lesions of symptomatic and asymptomatic teeth with vital pulps; (ii) to quantify the amount of endotoxin present; and (iii) to associate the presence of endotoxins with the acute pulpal pain derived from the irreversible pulpitis. Two specimens of carious dentine were taken under aseptic conditions from symptomatic teeth with irreversible pulpitis (n = 9) and asymptomatic teeth with reversible pulpitis (n = 11). The first specimen was taken from a layer of superficial caries and the second from a deeper one. Sound dentine was also collected from intact teeth without restoration and used as a noncarious control group (n = 4). During the patient sampling procedure an effort was made to collect an equal quantity of caries and sound dentine in all cases (approximately 6 mg). The extraction of endotoxins was performed using the Phenol-water method. The assay and quantitative determination of endotoxins was performed by the Quantitative Chromogenic test using Limulus Lysate. Data were analysed statistically using either independent or paired t-tests. The results indicated that endotoxins were present in the superficial and deep layers of caries of all symptomatic teeth with irreversible pulpitis (0.15078 and 0.12111 ng mL-1, respectively), with significantly greater amount (P reversible pulpitis (0.12091 and 0.07163 ng mL-1, respectively), with significantly greater amounts (P < 0.001) in the superficial compared to the deep layer. The results also demonstrated that significantly greater concentrations (P < 0.005) of endotoxins were present in the superficial carious layer of symptomatic compared with asymptomatic teeth (0.15078 and 0.12091 ng mL-1, respectively). Likewise, significantly greater amounts (P < 0.001) of endotoxins were present in the deep carious layer of symptomatic compared with asymptomatic teeth (0.12111 and 0.07163 ng mL-1

  3. Some metabolic effects of bacterial endotoxins in salmonid fishes

    Science.gov (United States)

    Wedemeyer, G.A.; Ross, A.J.; Smith, L.

    1968-01-01

    Coho salmon (Oncorhynchus kisutch) and rainbow trout (Salmo gairdneri) were highly resistant to endotoxins from both Escherichia coli and Aeromonas salmonicida (a fish pathogen) at 14 and 18 C.This resistance was investigated with liver tryptophan pyrrolase, liver glycogen depletion in vitro, and the arterial blood pressure as indicators. Liver glycogen depletion was accelerated by both endotoxins, but there was no significant cardiovascular response or effect on liver tryptophan pyrrolase activity. Since the cardiovascular effects of histamine were also limited, it was concluded that the metabolic effects of bacterial endotoxins in salmonids are qualitatively different from those of the higher vertebrates.

  4. Continuous blood purification treatment for endotoxin-induced acute respiratory distress syndrome.

    Science.gov (United States)

    Jiang, Y; Lin, R; Xu, Y; Zhang, S; Cui, K; Zhu, M; Li, A; Chen, C; Yang, J; Yang, W

    2017-02-16

    This study aimed to explore the effects of continuous blood purification (CBP) treatment in pigs affected with acute respiratory distress syndrome (ARDS). A total of 12 healthy male pigs, weighing 12±1.8 kg, were randomly and equally assigned to the control and experimental groups. The ARDS pig model was prepared by intravenous injections of endotoxin (20 µg/kg). The control group was given conventional supportive therapy, while the experimental group was given continuous veno-venous hemofiltration therapy. During the treatment process, the variations in dynamic lung compliance, oxygenation index, hemodynamics, and urine volume per hour at different times (Baseline, 0, 2, 4, and 6 h) were recorded. The levels of tumor necrosis factor (TNF-α), interleukin 6 (IL-6), and IL-10 in serum and bronchoalveolar lavage fluid (BALF) were measured using the enzyme-linked immunosorbent assay. The histomorphological changes of the lung, heart, and kidney were visualized using a light microscope. The nuclear factor κB p65 protein content of the heart, lung, and kidney tissues was also detected using western blot. The experimental group outperformed the control group in both respiratory and hemodynamic events. CBP treatment cleared TNF-α, IL-6, and IL-10 partially from serum and BALF. The pathological examination of the heart, lung, and kidney tissues revealed that the injury was less severe in the experimental group. CBP treatment can improve the organ functions of pigs affected with endotoxin-induced ARDS and protect these organs to some extent.

  5. Endotoxins in surgical instruments of hip arthroplasty.

    Science.gov (United States)

    Goveia, Vania Regina; Mendoza, Isabel Yovana Quispe; Guimarães, Gilberto Lima; Ercole, Flavia Falci; Couto, Bráulio Roberto Gonçalves Marinho; Leite, Edna Marilea Meireles; Stoianoff, Maria Aparecida Resende; Ferreira, José Antonio Guimarães

    2016-01-01

    To investigate endotoxins in sterilized surgical instruments used in hip arthroplasties. A descriptive exploratory study conducted in a public teaching hospital. Six types of surgical instruments were selected, namely: acetabulum rasp, femoral rasp, femoral head remover, chisel box, flexible bone reamer and femoral head test. The selection was based on the analysis of the difficulty in removing bone and blood residues during cleaning. The sample was made up of 60 surgical instruments, which were tested for endotoxins in three different stages. The EndosafeTM Gel-Clot LAL (Limulus Amebocyte Lysate method) was used. There was consistent gel formation with positive analysis in eight instruments, corresponding to 13.3%, being four femoral rasps and four bone reamers. Endotoxins in quantity ≥0.125 UE/mL were detected in 13.3% of the instruments tested. Investigar endotoxinas em instrumentais cirúrgicos esterilizados empregados em artroplastias do quadril. Estudo exploratório, descritivo, desenvolvido em um hospital público de ensino. Foram selecionados seis tipos de instrumentais, a saber: raspa acetabular, raspa femural, saca-cabeça de fêmur, formão box, fresa de fêmur e cabeça de prova de fêmur. A seleção foi feita a partir da análise da dificuldade para a remoção de resíduos de sangue e osso durante a limpeza. A amostra foi constituída por 60 instrumentais cirúrgicos, que foram testados para endotoxinas em três momentos distintos. Foi utilizado o método de gel-clot pelo Limulus Amebócito Lisado (LAL) Endosafe(tm). Houve formação de gel consistente com análise positiva em oito instrumentais, o que corresponde a 13,3%, sendo quatro raspas de fêmur e quatro fresas de fêmur. Foram detectadas endotoxinas em quantidade ≥0,125 UE/mL em 13,3% dos instrumentais testados.

  6. TRANSLOCATION OF BACTERIA AND ENDOTOXIN IN ORGAN DONORS

    NARCIS (Netherlands)

    van Goor, Harry; Rosman, C; Kooi, K; Wubbels, GH; Bleichrodt, RP

    1994-01-01

    Objective: To determine if bacterial translocation and endotoxin absorption occur in organ donors with an anatomically intact gastrointestinal tract. Design: Case series. Setting: Intensive care units in general and university hospitals. Patients: Twenty-one (multiple) organ donors. Intervention:

  7. Variability in endotoxin exposure levels and consequences for exposure assessment

    NARCIS (Netherlands)

    Spaan, S.; Schinkel, J.; Wouters, I.M.; Preller, L.; Tielemans, E.; Nij, E.T.; Heederik, D.

    2008-01-01

    Objectives: Workers in many industries are exposed to endotoxins, which may cause adverse health effects. In exposure assessment, information about exposure variability is essential. However, variability in exposure has rarely been investigated for biological agents and more specifically for

  8. Effect of hyperbaric air on endotoxin from Bacteroides fragilis strains

    Czech Academy of Sciences Publication Activity Database

    Chmelař, D.; Kašíková, A.; Martineková, P.; Hájek, M.; Rozložník, M.; Brabec, Marek; Janečková, J.; Vobejdová, J.; Čižnár, I.

    Online: 13 November 2017 (2018) ISSN 0015-5632 Institutional support: RVO:67985807 Keywords : endotoxin * hyperbaric condition Subject RIV: EE - Microbiology, Virology Impact factor: 1.521, year: 2016

  9. The endotoxin-induced increase of cytokines is followed by an increase of cortisol relative to dehydroepiandrosterone (DHEA) in healthy male subjects.

    Science.gov (United States)

    Straub, R H; Schuld, A; Mullington, J; Haack, M; Schölmerich, J; Pollmächer, T

    2002-11-01

    Dehydroepiandrosterone (DHEA) and DHEA sulphate (DHEAS) inhibit T-helper lymphocyte type 2 immune reactions and exert anti-inflammatory effects in some chronic inflammatory diseases. Both DHEA and, in particular, DHEAS levels are dramatically decreased in chronic inflammatory diseases whereas cortisol levels remain stable or are elevated. However, the time course of cortisol relative to DHEA production is not known. We tested whether administration of endotoxin to healthy male subjects can induce an early predominance of cortisol relative to DHEA and DHEAS. It is demonstrated that endotoxin induces a dose-dependent increase of cortisol in relation to DHEA (no effect at 0.2 ng endotoxin/kg body weight (b.w.), clear effect at 0.4 and 0.8 ng/kg b.w., pDHEAS (tested at 0.4 ng/kg b.w., P=0.014). The increase of cortisol relative to DHEA appears 4 h after endotoxin injection and 2 h after a strong increase of interleukin (IL)-6 relative to tumour necrosis factor (TNF). In addition, an increase of cortisol relative to 17OH-progesterone was observed. The ratio of serum IL-6/TNF was positively correlated with the ratio of serum cortisol/DHEA (R(Rank)=0.472, P=0.041) and serum cortisol/17OH-progesterone (R(Rank)=0.514, P=0.048). In conclusion, dissociation of cortisol relative to DHEA, DHEAS or 17OH-progesterone appears very early during a systemic inflammatory response which is associated with an increase of IL-6 relative to TNF. As in chronic inflammatory diseases, during an acute inflammatory response with endotoxin, these physiological hormone changes are probably necessary to achieve adequate cortisol levels at the expense of adrenal androgens.

  10. Elevated Levels and Different Repertoire Profile of Colostral Anti‐LPS Antibodies May Have a Significant Role in Compensating Newborn Immunity

    National Research Council Canada - National Science Library

    Nagao, A. T; Friedlander‐Del Nero, D; Arslanian, C; Carneiro‐Sampaio, M. M. S

    2001-01-01

    .... In the present study, we analyzed anti‐LPS O111 antibody isotypes in maternal serum and colostrum from mothers and in cord serum from their respective full‐term ( n  = 30) and preterm ( n  = 13) neonate infants...

  11. Metformin reduces the endotoxin-induced down-regulation of apolipoprotein E gene expression in macrophages

    Energy Technology Data Exchange (ETDEWEB)

    Stavri, Simona; Trusca, Violeta G.; Simionescu, Maya; Gafencu, Anca V., E-mail: anca.gafencu@icbp.ro

    2015-05-29

    The atheroprotective role of macrophage-derived apolipoprotein E (apoE) is well known. Our previous reports demonstrated that inflammatory stress down-regulates apoE expression in macrophages, aggravating atherogenesis. Metformin, extensively used as an anti-diabetic drug, has also anti-inflammatory properties, and thus confers vascular protection. In this study, we questioned whether metformin could have an effect on apoE expression in macrophages in normal conditions or under lipopolysaccharide (LPS)-induced stress. The results showed that metformin slightly increases the apoE expression only at high doses (5–10 mM). Low doses of metformin (1–3 mM) significantly reduce the LPS down-regulatory effect on apoE expression in macrophages. Our experiments demonstrated that LPS-induced NF-κB binds to the macrophage-specific distal regulatory element of apoE gene, namely to the multienhancer 2 (ME.2) and its 5′-deletion fragments. The NF-κB binding on ME.2 and apoE promoter has a down-regulatory effect. In addition, data revealed that metformin impairs NF-κB nuclear translocation, and thus, improves the apoE levels in macrophages under inflammatory stress. The positive effect of metformin in the inflammatory states, its clinical safety and low cost, make this drug a potential adjuvant in the therapeutic strategies for atherosclerosis. - Highlights: • High doses of metformin slightly increase apoE expression in macrophages. • Low doses of metformin up-regulate apoE gene in endotoxin-stressed macrophages. • Metformin reduces the negative effect of LPS on apoE expression by NF-κB inhibition.

  12. Human lipoproteins have divergent neutralizing effects on E. coli LPS, N. meningitidis LPS, and complete Gram-negative bacteria.

    Science.gov (United States)

    Sprong, Tom; Netea, Mihai G; van der Ley, Peter; Verver-Jansen, Trees J G; Jacobs, Liesbeth E H; Stalenhoef, Anton; van der Meer, Jos W M; van Deuren, Marcel

    2004-04-01

    The use of lipoproteins has been suggested as a treatment for Gram-negative sepsis because they inhibit lipopolysaccharide (LPS)-mediated cytokine production. However, little is known about the neutralizing effects of lipoproteins on cytokine production by meningococcal LPS or whole Gram-negative bacteria. We assessed the neutralizing effect of LDLs, HDLs, and VLDLs on LPS- or whole bacteria-induced cytokines in human mononuclear cells. A strong inhibition of Escherichia coli LPS-induced interleukin-1beta (IL-1beta), tumor necrosis factor-alpha, and IL-10 by LDL and HDL was seen, whereas VLDL had a less pronounced effect. In contrast, Neisseria meningitidis LPS, in similar concentrations, was neutralized much less effectively than E. coli LPS. Effective neutralization of meningococcal LPS required a longer interaction time, a lower concentration of LPS, or higher concentrations of lipoproteins. The difference in neutralization was independent of the saccharide tail, suggesting that the lipid A moiety accounted for the difference. Minimal neutralizing effects of the lipoproteins were observed on whole E. coli or N. meningitidis bacteria under all conditions tested. These results indicate that efficient neutralization of LPS depends on the type of LPS, but a sufficiently long interaction time, a low LPS concentration, or high lipoprotein concentration also inhibited cytokines by the less efficiently neutralized N. meningitidis LPS. Irrespective of these differences, whole bacteria showed no neutralization by lipoproteins.

  13. Protective effect of chlorpromazine on endotoxin toxicity and TNF production in glucocorticoid-sensitive and glucocorticoid-resistant models of endotoxic shock

    OpenAIRE

    1991-01-01

    The present study was designed to define the potential of chlorpromazine (CPZ) as a protective agent against lipopolysaccharide (LPS) toxicity in comparison with glucocorticoids, and to obtain initial correlations with its effects on the levels of tumor necrosis factor (TNF), a pivotal mediator of endotoxic shock. It was found that CPZ protects mice, normal or adrenalectomized, and guinea pigs against lethality of LPS, and inhibited TNF serum levels, like dexamethasone (DEX), a well-known inh...

  14. Peptidylprolyl cis/trans isomerase, NIMA-interacting 1 (PIN1) regulates pulmonary effects of endotoxin and tumor necrosis factor-α in mice.

    Science.gov (United States)

    Liu, Tongzheng; Schneider, Ryan A; Lee, Nam Y; Hoyt, Dale G

    2014-09-26

    Peptidylprolyl cis/trans isomerase, NIMA-interacting 1 (PIN1) modulates phospho-signaling by catalyzing rotation of the bond between a phosphorylated serine or threonine before proline in proteins. As depletion of PIN1 increased inflammatory protein expression in cultured endothelial cells treated with bacterial endotoxin (lipopolysaccharide, LPS) and interferon-γ, we hypothesized that PIN1 knockout would increase sensitivity to LPS-induced lung inflammation in mice. Mortality due to a high dose of LPS (30mg/kg) was greater in knockout than wildtype mice. Lung myeloperoxidase activity, reflecting neutrophils, was increased to a 35% higher level in PIN1 knockout mouse lung, as compared with wildtype, after treatment with a sublethal dose of 3mgLPS/kg, ip. Unexpectedly, plasma tumor necrosis factor-α (TNF) was approximately 50% less than in wildtype mice. Knockout mice, however, were more sensitive than wildtype to TNF-induced neutrophil accumulation. The neutrophil adhesion molecule, E-selectin, was also elevated in lungs of knockout mice treated with TNF, suggesting that PIN1 depletion increases endothelial sensitivity to TNF. Indeed, TNF induced more reactive oxygen species in cultured endothelial cells depleted of PIN1 with short hairpin RNA than in control cells. Collectively, the results indicate that while PIN1 normally facilitates TNF production in LPS-treated mice, it suppresses pulmonary and endothelial reactions to the cytokine. Tissue or cell-specific effects of PIN1 may affect the overall inflammatory response to LPS and other stimuli. Copyright © 2014 Elsevier Inc. All rights reserved.

  15. Endotoxins-the invisible companion in biomaterials research.

    Science.gov (United States)

    Lieder, Ramona; Petersen, Pétur Henry; Sigurjónsson, Ólafur Eysteinn

    2013-10-01

    Metal implants and polymeric devices for the application in the clinical treatment of orthopedic tissue injuries are increasingly coated with bioactive biomaterials derived from natural substances to induce desirable biological effects. Many metals and polymers used in biomaterials research show high affinity for endotoxins, which are abundant in the environment. Endotoxin contamination is indicated in the pathology of periodontitis and aseptic implant loosening, but may also affect the evaluation of a biomaterial's bioactivity by inducing strong inflammatory reactions. In this review, we discuss the high affinity of three commonly used implant biomaterials for endotoxins and how the contamination can affect the outcome of the orthopedic fixation. The chemical nature of bacterial endotoxins and some of the clinical health implications are described, as this knowledge is critically important to tackle the issues associated with the measurement and removal of endotoxins from medical devices. Commonly used methods for endotoxin testing and removal from natural substances are examined and the lack of standard guidelines for the in vitro evaluation of biomaterials is discussed.

  16. Salidroside attenuates LPS-induced pro-inflammatory cytokine responses and improves survival in murine endotoxemia.

    Science.gov (United States)

    Guan, Shuang; Feng, Haihua; Song, Bocui; Guo, Weixiao; Xiong, Ying; Huang, Guoren; Zhong, Weiting; Huo, Meixia; Chen, Na; Lu, Jing; Deng, Xuming

    2011-12-01

    Salidroside is a major component isolated from the Rhodiola rosea. In the present study, we investigated the anti-inflammatory effects of salidroside on cytokine production by lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages in vitro, and the results showed that salidroside reduced tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) secretions. This inspired us to further study the effects of salidroside in vivo. Salidroside significantly attenuated TNF-α, IL-1β and IL-6 productions in serum from mice challenged with LPS, and consistent with the results in vitro. In the murine model of endotoxemia, mice were treated with salidroside prior to or after LPS challenge. The results showed that salidroside significantly increased mouse survival. Further studies revealed that salidroside could downregulate LPS-induced nuclear transcription factor-қB (NF-қB) DNA-binding activation and ERK/MAPKs signal transduction pathways production in RAW 264.7 macrophages. These observations indicated that salidroside modulated early cytokine responses by blocking NF-қB and ERK/MAPKs activation, and thus, increased mouse survival. These effects of salidroside may be of potential usefulness in the treatment of inflammation-mediated endotoxemia. Copyright © 2011 Elsevier B.V. All rights reserved.

  17. PEGylated porcine glucagon-like peptide-2 improved the intestinal digestive function and prevented inflammation of weaning piglets challenged with LPS.

    Science.gov (United States)

    Qi, K K; Wu, J; Deng, B; Li, Y M; Xu, Z W

    2015-09-01

    This study was conducted to determine the effects on intestinal function, anti-inflammatory role and possible mechanism of polyethylene glycosylated (PEGylated) porcine glucagon-like peptide-2 (pGLP-2), a long-acting form of pGLP-2, in weaning piglets challenged with Escherichia coli lipopolysaccharide (LPS). We divided 18 weaned piglets on day 21 into three groups (control, LPS and LPS+PEG-pGLP-2; n=6). The piglets from the LPS+PEG-pGLP-2 group were injected with PEG-pGLP-2 at 10 nmol/kg BW from 5 to 7 days of the trials daily. On 8th day, the piglets in the LPS and LPS+PEG-pGLP-2 groups were intraperitoneally administered with 100 µg LPS/kg. The control group was administered with the same volume of saline solution. The piglets were then sacrificed on day 28. Afterwards, serum, duodenum, jejunum and ileum samples were collected for analysis of structural and functional endpoints. LPS+PEG-pGLP-2 treatment increased (Pdigestive function were associated with the release of GLP-2R mediator (keratinocyte growth factor) and the decrease in the expressions of intestinal pro-inflammatory cytokines.

  18. Anti-inflammatory activity of cinnamon water extract in vivo and in vitro LPS-induced models.

    Science.gov (United States)

    Hong, Joung-Woo; Yang, Ga-Eun; Kim, Yoon Bum; Eom, Seok Hyun; Lew, Jae-Hwan; Kang, Hee

    2012-11-28

    Cinnamon bark is one of the most popular herbal ingredients in traditional oriental medicine and possesses diverse pharmacological activities including anti-bacterial, anti-viral, and anti-cancer properties. The goal of this study is to investigate the in vivo and in vitro inhibitory effect of cinnamon water extract (CWE) on lipopolysaccharide (LPS)-induced tumor necrosis factor (TNF)-α and its underlying intracellular mechanisms. CWE was orally administrated to mice for 6 days prior to intraperitoneal injection of LPS. Serum levels of TNF-α and interleukin (IL)-6 were determined 1 hour after LPS stimulation. Peritoneal macrophages from thioglycollate-injected mice were isolated and assayed for viability, cytokine expression and signaling molecules upon LPS stimulation. CWE was further fractioned according to molecular size, and the levels of total polyphenols and biological activities of each fraction were measured. The oral administration of CWE to mice significantly decreased the serum levels of TNF-α and IL-6. CWE treatment in vitro decreased the mRNA expression of TNF-α. CWE blocked the LPS-induced degradation of IκBα as well as the activation of JNK, p38 and ERK1/2. Furthermore, size-based fractionation of CWE showed that the observed inhibitory effect of CWE in vitro occurred in the fraction containing the highest level of total polyphenols. Treatment with CWE decreased LPS-induced TNF-α in serum. In vitro inhibition of TNF-α gene by CWE may occur via the modulation of IκBα degradation and JNK, p38, and ERK1/2 activation. Our results also indicate that the observed anti-inflammatory action of CWE may originate from the presence of polyphenols.

  19. Anti-inflammatory activity of cinnamon water extract in vivo and in vitro LPS-induced models

    Directory of Open Access Journals (Sweden)

    Hong Joung-Woo

    2012-11-01

    Full Text Available Abstract Background Cinnamon bark is one of the most popular herbal ingredients in traditional oriental medicine and possesses diverse pharmacological activities including anti-bacterial, anti-viral, and anti-cancer properties. The goal of this study is to investigate the in vivo and in vitro inhibitory effect of cinnamon water extract (CWE on lipopolysaccharide (LPS-induced tumor necrosis factor (TNF-α and its underlying intracellular mechanisms. Methods CWE was orally administrated to mice for 6 days prior to intraperitoneal injection of LPS. Serum levels of TNF-α and interleukin (IL-6 were determined 1 hour after LPS stimulation. Peritoneal macrophages from thioglycollate-injected mice were isolated and assayed for viability, cytokine expression and signaling molecules upon LPS stimulation. CWE was further fractioned according to molecular size, and the levels of total polyphenols and biological activities of each fraction were measured. Results The oral administration of CWE to mice significantly decreased the serum levels of TNF-α and IL-6. CWE treatment in vitro decreased the mRNA expression of TNF-α. CWE blocked the LPS-induced degradation of IκBα as well as the activation of JNK, p38 and ERK1/2. Furthermore, size-based fractionation of CWE showed that the observed inhibitory effect of CWE in vitro occurred in the fraction containing the highest level of total polyphenols. Conclusions Treatment with CWE decreased LPS-induced TNF-α in serum. In vitro inhibition of TNF-α gene by CWE may occur via the modulation of IκBα degradation and JNK, p38, and ERK1/2 activation. Our results also indicate that the observed anti-inflammatory action of CWE may originate from the presence of polyphenols.

  20. Fractal-like kinetics of intracellular enzymatic reactions: a chemical framework of endotoxin tolerance and a possible non-specific contribution of macromolecular crowding to cross-tolerance

    Science.gov (United States)

    2013-01-01

    Background The response to endotoxin (LPS), and subsequent signal transduction lead to the production of cytokines such as tumor necrosis factor-α (TNF-α) by innate immune cells. Cells or organisms pretreated with endotoxin enter into a transient state of hyporesponsiveness, referred to as endotoxin tolerance (ET) which represents a particular case of negative preconditioning. Despite recent progress in understanding the molecular basis of ET, there is no consensus yet on the primary mechanism responsible for ET and for the more complex cases of cross tolerance. In this study, we examined the consequences of the macromolecular crowding (MMC) and of fractal-like kinetics (FLK) of intracellular enzymatic reactions on the LPS signaling machinery. We hypothesized that this particular type of enzyme kinetics may explain the development of ET phenomenon. Method Our aim in the present study was to characterize the chemical kinetics framework in ET and determine whether fractal-like kinetics explains, at least in part, ET. We developed an ordinary differential equations (ODE) mathematical model that took into account the links between the MMC and the LPS signaling machinery leading to ET. We proposed that the intracellular fractal environment (MMC) contributes to ET and developed two mathematical models of enzyme kinetics: one based on Kopelman’s fractal-like kinetics framework and the other based on Savageau’s power law model. Results Kopelman’s model provides a good image of the potential influence of a fractal intracellular environment (MMC) on ET. The Savageau power law model also partially explains ET. The computer simulations supported the hypothesis that MMC and FLK may play a role in ET. Conclusion The model highlights the links between the organization of the intracellular environment, MMC and the LPS signaling machinery leading to ET. Our FLK-based model does not minimize the role of the numerous negative regulatory factors. It simply draws attention to

  1. Pantoea agglomerans: a mysterious bacterium of evil and good. Part II. Deleterious effects: Dust-borne endotoxins and allergens – focus on grain dust, other agricultural dusts and wood dust

    Directory of Open Access Journals (Sweden)

    Jacek Dutkiewicz

    2015-12-01

    Full Text Available [i][/i][i]Pantoea agglomerans[/i], a Gram-negative bacterium developing in a variety of plants as epiphyte or endophyte is particularly common in grain and grain dust, and has been identified by an interdisciplinary group from Lublin, eastern Poland, as a causative agent of work-related diseases associated with exposure to grain dust and other agricultural dusts. The concentration of [i]P. agglomerans[/i] in grain as well as in the settled grain and flour dust was found to be high, ranging from 10[sup]4[/sup] –10[sup]8 [/sup] CFU/g, while in the air polluted with grain or flour dust it ranged from 10[sup]3[/sup] –10[sup]5[/sup] CFU/m[sup]3[/sup] and formed 73.2–96% of the total airborne Gram-negative bacteria. The concentration of [i]P. agglomerans[/i] was also relatively high in the air of the facilities processing herbs and other plant materials, while it was lower in animal farms and in wood processing facilities. [i]Pantoea agglomerans[/i] produces a biologically-potent endotoxin (cell wall lipopolysaccharide, LPS. The significant part of this endotoxin occurs in dusts in the form of virus-sized globular nanoparticles measuring 10–50 nm that could be described as the ‘endotoxin super-macromolecules’. A highly significant relationship was found (R=0.804, P=0.000927 between the concentration of the viable [i]P. agglomerans[/i] in the air of various agricultural and wood industry settings and the concentration of bacterial endotoxin in the air, as assessed by the Limulus test. Although this result may be interfered by the presence of endotoxin produced by other Gram-negative species, it unequivocally suggests the primary role of the [i]P. agglomerans[/i] endotoxin as an adverse agent in the agricultural working environment, causing toxic pneumonitis (ODTS. Numerous experiments by the inhalation exposure of animals to various extracts of [i]P. agglomerans[/i] strains isolated from grain dust, including endotoxin isolated with

  2. Pantoea agglomerans: a mysterious bacterium of evil and good. Part II--Deleterious effects: Dust-borne endotoxins and allergens--focus on grain dust, other agricultural dusts and wood dust.

    Science.gov (United States)

    Dutkiewicz, Jacek; Mackiewicz, Barbara; Lemieszek, Marta Kinga; Golec, Marcin; Skórska, Czesława; Góra-Florek, Anna; Milanowski, Janusz

    2016-01-01

    Pantoea agglomerans, a Gram-negative bacterium developing in a variety of plants as epiphyte or endophyte is particularly common in grain and grain dust, and has been identified by an interdisciplinary group from Lublin, eastern Poland, as a causative agent of work-related diseases associated with exposure to grain dust and other agricultural dusts. The concentration of P. agglomerans in grain as well as in the settled grain and flour dust was found to be high, ranging from 10(4)-10(8) CFU/g, while in the air polluted with grain or flour dust it ranged from 10(3)-10(5) CFU/m(3) and formed 73.2-96% of the total airborne Gram-negative bacteria. The concentration of P. agglomerans was also relatively high in the air of the facilities processing herbs and other plant materials, while it was lower in animal farms and in wood processing facilities. Pantoea agglomerans produces a biologically-potent endotoxin (cell wall lipopolysaccharide, LPS). The significant part of this endotoxin occurs in dusts in the form of virus-sized globular nanoparticles measuring 10-50 nm that could be described as the 'endotoxin super-macromolecules'. A highly significant relationship was found (R=0.804, P=0.000927) between the concentration of the viable P. agglomerans in the air of various agricultural and wood industry settings and the concentration of bacterial endotoxin in the air, as assessed by the Limulus test. Although this result may be interfered by the presence of endotoxin produced by other Gram-negative species, it unequivocally suggests the primary role of the P. agglomerans endotoxin as an adverse agent in the agricultural working environment, causing toxic pneumonitis (ODTS). Numerous experiments by the inhalation exposure of animals to various extracts of P. agglomerans strains isolated from grain dust, including endotoxin isolated with trichloroacetic acid (LPS-TCA), endotoxin nanoparticles isolated in sucrose gradient (VECN), and mixture of proteins and endotoxin obtained

  3. Endotoxin, ergosterol, fungal DNA and allergens in dust from schools in Johor Bahru, Malaysia- associations with asthma and respiratory infections in pupils.

    Directory of Open Access Journals (Sweden)

    Dan Norbäck

    Full Text Available There are few studies on associations between respiratory health and allergens, fungal and bacterial compounds in schools in tropical countries. The aim was to study associations between respiratory symptoms in pupils and ethnicity, chemical microbial markers, allergens and fungal DNA in settled dust in schools in Malaysia. Totally 462 pupils (96% from 8 randomly selected secondary schools in Johor Bahru, Malaysia, participated. Dust was vacuumed from 32 classrooms and analysed for levels of different types of endotoxin as 3-hydroxy fatty acids (3-OH, muramic acid, ergosterol, allergens and five fungal DNA sequences. Multiple logistic regression was applied. Totally 13.1% pupils reported doctor's diagnosed asthma, 10.3% wheeze and 21.1% pollen or pet allergy. Indian and Chinese children had less atopy and asthma than Malay. Carbon dioxide levels were low (380-690 ppm. No cat (Fel d1, dog (Can f 1 or horse allergens (Ecu cx were detected. The levels of Bloomia tropicalis (Blo t, house dust mite allergens (Der p 1, Der f 1, Der m 1 and cockroach allergens (Per a 1 and Bla g 1 were low. There were positive associations between levels of Aspergillus versicolor DNA and daytime breathlessness, between C14 3-OH and respiratory infections and between ergosterol and doctors diagnosed asthma. There were negative (protective associations between levels of C10 3-OH and wheeze, between C16 3-OH and day time and night time breathlessness, between cockroach allergens and doctors diagnosed asthma. Moreover there were negative associations between amount of fine dust, total endotoxin (LPS and respiratory infections. In conclusion, endotoxin at school seems to be mainly protective for respiratory illness but different types of endotoxin could have different effects. Fungal contamination measured as ergosterol and Aspergillus versicolor DNA can be risk factors for respiratory illness. The ethnical differences for atopy and asthma deserve further attention.

  4. Upregulation of ICAM-1 Expression on J774.2 Macrophages by Endotoxin Involves Activation of NF-κB but not Protein Tyrosine Kinase: Comparison to Induction of iNOS

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    Hartmut Ruetten

    1999-01-01

    Full Text Available This study compares the signal transduction pathway which leads to the upregulation of intercellular adhesion molecule-1 (ICAM-1 expression with that of the increase in the expression of inducible nitric oxide synthase (iNOS protein and activity caused by endotoxin in cultured J774.2 macrophages. Treatment of J774.2 cells with lipopolysaccharide E. coli (LPS induced a concentration-dependent increase in the expression of ICAM-1 on the cell surface within 4 h and an increase in iNOS protein and activity at 24 h. The upregulation of ICAM-1 expression on J774.2 macrophages caused by LPS was significantly inhibited by pretreatment of the cells with inhibitors of the activation of the nuclear transcription factor NF-κB, such as L-1-tosylamido-2-phenylethylchloromethyl ketone (TPCK, pyrrolidine dithiocarbamate (PDTC, rotenone or calpain inhibitor I, but not by the tyrosine kinase inhibitors, tyrphostin AG126 or genistein. In contrast, genistein or tyrphostin AG126 also prevented the induction of iNOS protein and activity in J774.2 macrophages elicited by LPS. Thus, the increase in the expression of ICAM-1 on J774.2 macrophages by endotoxin involves the activation of NFκB, but not of protein tyrosine kinase.

  5. Analysis of endotoxin and endothelin-1 levels in patients with type 1 hepatorenal syndrome

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    GAO Baoxiu

    2014-01-01

    Full Text Available ObjectiveTo analyze the clinical data, laboratory parameters, infection rate, and serum procalcitonin (PCT and ET-1 levels of patients with cirrhotic ascites and type 1 hepatorenal syndrome (HRS and to investigate the roles of endotoxin and ET-1 in the development of HRS. MethodsBetween January 2009 and October 2012, 56 inpatients with cirrhotic ascites and type 1 HRS (HRS group and 60 inpatients with cirrhotic ascites who had normal renal function (non-HRS group were included in the study. Their general data, causes of liver cirrhosis, infection rates and types, Child-Pugh classification, systemic inflammatory response syndrome (SIRS score, and mean arterial pressure (MAP were recorded; blood samples were collected to evaluate liver and renal function and measure serum electrolyte, PCT, and ET-1 levels. The clinical data and laboratory parameters were compared between the two groups. Categorical data were analyzed by chi-square test; comparison of normally distributed continuous data between the two groups was made by independent-samples t test, and comparison of non-normally distributed continuous data between the two groups was made by Wilcoxon rank sum test. ResultsThe infection rate of HRS group (75.0% was significantly higher than that of non-HRS group (28.4% (χ2=11.91, P<0.05. The PCT and ET-1 levels and SIRS score of HRS group [8.72 (3.14, 31.68 ng/L, 13.04±2.82 pg/ml, and 2.1±1.1] were significantly higher than those of non-HRS group [0.11 (0.04, 0.45 ng/L, 5.76±1.68 pg/ml, and 0.6±0.6] (P<0.05. In addition, the HRS group had significantly higher serum urea, creatine, cystatin C, and K levels than the non-HRS group (P<0.05, while the HRS group had significantly lower Na and Cl levels than the non-HRS group (P<0.05. There were no significant differences in ALT and AST levels between the two groups (P>005. ConclusionEndotoxin causes elevated expression of ET-1, and ET-1 induces renal perfusion deficiency by

  6. Autodisplay of the La/SSB protein on LPS-free E. coli for the diagnosis of Sjögren's syndrome.

    Science.gov (United States)

    Yoo, Gu; Dilkaute, Carina; Bong, Ji-Hong; Song, Hyun-Woo; Lee, Misu; Kang, Min-Jung; Jose, Joachim; Pyun, Jae-Chul

    2017-05-01

    The objective of this study was to present an immunoassay for the diagnosis of Sjögren's syndrome based on the autodisplayed La/SSB protein on the outer membrane of intact E. coli (strain UT-5600) and LPS-free E. coli (ClearColi™). As the first step, an autodisplay vector (pCK002) was transfected into intact E. coli and LPS-free E. coli for comparison of efficiency of autdisplay of La/SSB. The maximal level of La/SSB expression was estimated to be similar for LPS-free E. coli and intact E. coli at different optimal induction periods. Intact E. coli was found to grow twofold faster than LPS-free E. coli, and the maximal level of expression for LPS-free E. coli was obtained with a longer induction period. When the zeta potential was measured, both intact E. coli and LPS-free E. coli showed negative values, and the autodisplay of negatively charged La/SSB protein (pIE. coli and LPS-free E. coli resulted in a slight change in zeta potential values. E. coli with autodisplayed La/SSB protein was used for an immunoassay of anti-La/SSB antibodies for the diagnosis of Sjögren's syndrome. The surface of E. coli with the autodisplayed antigen was modified with rabbit serum and papain to prevent false positive signals because of nonspecific binding of unrelated antibodies from human serum. LPS-free E. coli with autodisplayed La/SSB protein yielded sensitivity and selectivity of 81.6% and 78.6%, respectively. The Bland-Altman test showed that the immunoassays based on LPS-free E. coli and intact E. coli with autodisplayed La/SSB protein were statistically equivalent to a clinical immunoassay for detection of anti-La/SSB antibodies (confidence coefficient 95%). Copyright © 2017 Elsevier Inc. All rights reserved.

  7. Relationships between metabolic changes and clinical signs in pregnant sheep given endotoxin.

    OpenAIRE

    Naylor, J M; Kronfeld, D. S.

    1986-01-01

    Groups of four pregnant ewes were allocated to the following feeding and intravenous endotoxin treatments: fed, Escherichia coli endotoxin (50 micrograms/kg X 75), fed, saline, fasted, E. coli endotoxin (50 micrograms/kg X 75) and fasted, saline. Endotoxin administration resulted in depression, fever, hypoglycemia, hypocalcemia and a reduction in nonesterified fatty acid and ketone body concentrations. Depression correlated best with body temperature (r = 0.76), fasted sheep showed smaller in...

  8. Exposure to airborne endotoxins among sewer workers: an exploratory study.

    Science.gov (United States)

    Duquenne, Philippe; Ambroise, Denis; Görner, Pierre; Clerc, Frédéric; Greff-Mirguet, Guylaine

    2014-04-01

    Exploratory bioaerosol sampling was performed in order to assess exposure to airborne endotoxins during sewer work. Personal samples were collected in underground sewer pipes using 37-mm closed-face cassettes containing fibreglass filters (CFC-FG method) or polycarbonate filters (CFC-PC method). Endotoxins were quantified using the limulus amoebocyte lysate assay. Concentrations of airborne endotoxins at sewer workplaces (16-420 EU m(-3)) were higher than those measured outside the sewer network (0.6-122 EU m(-3)). Sewer worker exposure to airborne endotoxins depended on the workplace and on the tasks. Exposure levels were the highest for tasks involving agitation of water and matter, especially for 'chamber cleanup' and 'pipes cleanup' with a high-pressure water jet. Airborne endotoxin levels at the workplace tended to be higher when CFC-FG was used as the sampling method rather than CFC-PC. The adjusted mean of the measured concentrations for CFC-PC represents 57% of the mean observed with CFC-FG. The number of samples collected in the descriptive study was too low for drawing definitive conclusions and further exposure investigations are needed. Therefore, our exploratory study provides new exposure data for the insufficiently documented sewer working environment and it would be useful for designing larger exposures studies.

  9. Serum sickness

    Science.gov (United States)

    Drug allergy - serum sickness; Allergic reaction - serum sickness; Allergy - serum sickness ... symptoms of serum sickness. Certain medicines (such as penicillin, cefaclor, and sulfa) can cause a similar reaction. ...

  10. SORPTION OF BACTERIAL-ENDOTOXIN AND RETENTION OF BACTERIA BY POSITIVELY CHARGED MEMBRANE FILTERS

    NARCIS (Netherlands)

    VANDOORNE, H

    1993-01-01

    Positively charged membrane filters (Pall, Posidyne(R)) were studied for their ability to retain both bacterial endotoxins and bacteria. Filters (0.2 mum pore size) were challenged with high (up to 5*10(5) EU) levels of bacterial endotoxin. When the endotoxin was dispersed in water, a more than

  11. Airborne endotoxin concentrations at a large open-lot dairy in southern Idaho

    Science.gov (United States)

    Endotoxins are derived from Gram-negative bacteria and are a potential respiratory health risk for animals and humans. To determine the potential for endotoxin transport from a large open lot dairy, airborne endotoxin concentrations were determined at an upwind location (background) and five downwi...

  12. 40 CFR 180.1107 - Delta endotoxin of Bacillus thuringiensis variety kurstaki encapsulated into killed Pseudomonas...

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Delta endotoxin of Bacillus... From Tolerances § 180.1107 Delta endotoxin of Bacillus thuringiensis variety kurstaki encapsulated into killed Pseudomonas fluorescens; exemption from the requirement of a tolerance. The delta endotoxin of...

  13. Endotoxin-induced antitumor activity in the mouse is highly potentiated by muramyl dipeptide

    NARCIS (Netherlands)

    Hofhuis, F.M.A.; Bloksma, N.; Willers, J.M.N.

    1984-01-01

    The ability of aqueous solutions of various endotoxin preparations, muramyl dipeptide (MDP) and combinations of endotoxin and MDP, to induce necrosis and regression of subcutaneous Meth A transplants in mice and their toxicity were studied. While intravenously injected toxic endotoxins, in contrast

  14. Effect of enteral versus parenteral nutrition on inflammation and cardiac function in a rat model of endotoxin-induced sepsis.

    Science.gov (United States)

    Hagiwara, Satoshi; Iwasaka, Hideo; Shingu, Chihiro; Noguchi, Takayuki

    2008-09-01

    The present study examined whether total enteral nutrition (TEN) and total parenteral nutrition (TPN) differentially modulates ghrelin, inflammatory mediator production, and cardiac dysfunction induced by LPS. Rats received isocaloric parenteral or enteral nutrition through implanted vascular catheters or gastrostomy tubes for 7 days. Enteral nutrition was provided in conventional (TEN-C) and immunonutrition (TEN-I) formulations. Subsequently, rats were injected i.v. with LPS. Serum ghrelin, TNF-alpha, and high mobility group box 1 levels were determined by enzyme-linked immunosorbent assay. Myocardiac function was also assessed via the Langendorff isolated heart technique. The TEN-C increased plasma ghrelin and inhibited inflammatory mediators both before and after LPS administration when compared with TPN or TEN-I. Furthermore, animals receiving TPN and TEN-I had significantly lower left ventricular developed pressure but increased pressure development during isovolumetric contraction (dP/dt(max)) and relaxation (dP/dt(min)) when compared with animals receiving TEN-C after LPS-induced shock (P maintaining cardiac function during LPS-induced septic shock.

  15. Placental transfer of IgG antibodies specific to Klebsiella and Pseudomonas LPS and to group B Streptococcus in twin pregnancies.

    Science.gov (United States)

    Stach, S C L; Brizot, M L; Liao, A W; Palmeira, P; Francisco, R P V; Carneiro-Sampaio, M M S; Zugaib, M

    2015-02-01

    Group B Streptococcus (GBS), Klebsiella spp. and Pseudomonas spp. are important aetiological agents of neonatal infections in Brazil. There is a lack of data in the literature regarding the specific transport of immunoglobulin G (IgG) against these pathogens in multiple pregnancies. Maternal (n = 55) and umbilical cord (n = 110) blood samples were prospectively collected at birth from 55 twin pregnancies. The factors associated with cord levels and transfer ratios of IgG against GBS, Klebsiella and Pseudomonas were examined. The IgG umbilical cord serum levels specific to GBS, Klebsiella LPS and Pseudomonas LPS were significantly associated with maternal-specific IgG concentrations and the presence of diabetes. The anti-Klebsiella IgG cord serum concentrations were also related to birthweight and the presence of hypertension. The transfer ratios against GBS and Pseudomonas LPS were associated with maternal-specific IgG concentrations. The transfer ratios for GBS and Pseudomonas LPS were associated with gestational age at delivery and the presence of diabetes, respectively. None of the examined parameters were related to Klebsiella LPS transfer ratios. We conclude that in twin pregnancies, specific maternal IgG serum concentrations and diabetes were the parameters associated with umbilical cord serum IgG concentrations reactive with the three pathogens investigated. All the other parameters investigated showed different associations with neonatal-specific IgG levels according to the antigen studied. There was no uniformity of the investigated parameters regarding association with placental IgG transfer ratios against the GBS, Pseudomonas LPS and Klebsiella LPS. © 2014 John Wiley & Sons Ltd.

  16. Protective effect of chlorpromazine on endotoxin toxicity and TNF production in glucocorticoid-sensitive and glucocorticoid-resistant models of endotoxic shock

    Science.gov (United States)

    1991-01-01

    The present study was designed to define the potential of chlorpromazine (CPZ) as a protective agent against lipopolysaccharide (LPS) toxicity in comparison with glucocorticoids, and to obtain initial correlations with its effects on the levels of tumor necrosis factor (TNF), a pivotal mediator of endotoxic shock. It was found that CPZ protects mice, normal or adrenalectomized, and guinea pigs against lethality of LPS, and inhibited TNF serum levels, like dexamethasone (DEX), a well-known inhibitor of TNF synthesis. CPZ protected against LPS lethality when administered 30 minutes (min) before, simultaneously, or up to 10 min after LPS and was ineffective when given 30 min after LPS, paralleling the inhibitory effect on TNF production. In another experimental model, where mice were sensitized to LPS toxicity by actinomycin D, CPZ significantly inhibited LPS lethality and hepatotoxicity, whereas under these conditions DEX was inactive. These experiments indicate that CPZ has a protective action in both glucocorticoid-sensitive and -resistant models of endotoxic shock. PMID:2033366

  17. Astragaloside IV Inhibits NF-κB Activation and Inflammatory Gene Expression in LPS-Treated Mice

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    Wei-Jian Zhang

    2015-01-01

    Full Text Available In this study we investigated the role of astragaloside IV (AS-IV, one of the major active constituents purified from the Chinese medicinal herb Astragalus membranaceus, in LPS-induced acute inflammatory responses in mice in vivo and examined possible underlying mechanisms. Mice were assigned to four groups: vehicle-treated control animals; AS-IV-treated animals (10 mg/kg b.w. AS-IV daily i.p. injection for 6 days; LPS-treated animals; and AS-IV plus LPS-treated animals. We found that AS-IV treatment significantly inhibited LPS-induced increases in serum levels of MCP-1 and TNF by 82% and 49%, respectively. AS-IV also inhibited LPS-induced upregulation of inflammatory gene expression in different organs. Lung mRNA levels of cellular adhesion molecules, MCP-1, TNFα, IL-6, and TLR4 were significantly attenuated, and lung neutrophil infiltration and activation were strongly inhibited, as reflected by decreased myeloperoxidase content, when the mice were pretreated with AS-IV. Similar results were observed in heart, aorta, kidney, and liver. Furthermore, AS-IV significantly suppressed LPS-induced NF-κB and AP-1 DNA-binding activities in lung and heart. In conclusion, our data provide new in vivo evidence that AS-IV effectively inhibits LPS-induced acute inflammatory responses by modulating NF-κB and AP-1 signaling pathways. Our results suggest that AS-IV may be useful for the prevention or treatment of inflammatory diseases.

  18. Determination of endotoxins in sugar with the Limulus test.

    Science.gov (United States)

    Haskå, G; Nystrand, R

    1979-01-01

    The Limulus amebocyte lysate test has been used for determination of pyrogens in sugar of different qualities. All the samples of domestic white sugar and beet raw sugar produced in Sweden during 1976 had a very low content of endotoxins, less than 10 ng/g of sugar. Imported cane raw sugar was, however, highly contaminated. The highest value obtained corresponds to about 100 mg of Escherichia coli endotoxin per g of raw sugar. Such crude sugar cannot, even after refining, be used for medical purposes. Instead, Swedish beet sugar is used as the raw material for production of invert sugar solutions for parenteral administration. The amount of endotoxin in this sugar is less than 1 ng/g. PMID:526016

  19. Endotoxin, ergosterol, muramic acid and fungal DNA in dust from schools in Johor Bahru, Malaysia — Associations with rhinitis and sick building syndrome (SBS) in junior high school students

    Energy Technology Data Exchange (ETDEWEB)

    Norbäck, Dan, E-mail: dan.norback@medsci.uu.se [Department of Medical Science, Occupational and Environmental Medicine, Uppsala University Hospital, Uppsala University, Uppsala (Sweden); Hashim, Jamal Hisham [United Nations University—International Institute for Global Health, Kuala Lumpur (Malaysia); Department of Community Health, National University of Malaysia, Kuala Lumpur (Malaysia); Markowicz, Pawel [Division of Medical Microbiology, Department of Laboratory Medicine, University of Lund, Lund (Sweden); Cai, Gui-Hong [Department of Medical Science, Occupational and Environmental Medicine, Uppsala University Hospital, Uppsala University, Uppsala (Sweden); Hashim, Zailina [Department of Environmental and Occupational Health, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor (Malaysia); Ali, Faridah [Primary Care Unit, Johor State Health Department, Johor Bahru (Malaysia); Larsson, Lennart [Division of Medical Microbiology, Department of Laboratory Medicine, University of Lund, Lund (Sweden)

    2016-03-01

    This paper studied associations between ocular symptoms, rhinitis, throat and dermal symptoms, headache and fatigue in students by ethnicity and in relation to exposure to chemical microbial markers and fungal DNA in vacuumed dust in schools in Malaysia. A total of 462 students from 8 randomly selected secondary schools in Johor Bahru, Malaysia, participated (96% response rate). Dust was vacuumed from 32 classrooms and analysed for levels of five types of endotoxin as 3-hydroxy fatty acids (C10, C12, C14, C16 and C18 3-OH), muramic acid, ergosterol and five sequences of fungal DNA. Multiple logistic regression was applied. Totally 11.9% reported weekly ocular symptoms, 18.8% rhinitis, 15.6% throat and 11.1% dermal symptoms, 20.6% headache and 22.1% tiredness. Totally 21.1% reported pollen or furry pet allergy (atopy) and 22.0% parental asthma or allergy. Chinese students had less headache than Malay and Indian had less rhinitis and less tiredness than Malay. Parental asthma/allergy was a risk factor for ocular (odds ratio = 3.79) and rhinitis symptoms (OR = 3.48). Atopy was a risk factor for throat symptoms (OR = 2.66), headache (OR = 2.13) and tiredness (OR = 2.02). There were positive associations between amount of fine dust in the dust samples and ocular symptoms (p < 0.001) and rhinitis (p = 0.006). There were positive associations between C14 3-OH and rhinitis (p < 0.001) and between C18 3-OH and dermal symptoms (p = 0.007). There were negative (protective) associations between levels of total endotoxin (LPS) (p = 0.004) and levels of ergosterol (p = 0.03) and rhinitis and between C12 3-OH and throat symptoms (p = 0.004). In conclusion, the amount of fine dust in the classroom was associated with rhinitis and other SBS symptoms and improved cleaning of the schools is important. Endotoxin in the school dust seems to be mainly protective for rhinitis and throat symptoms but different types of endotoxin could have different effects. The ethnic differences in

  20. The anti-inflammatory and antioxidant effects of melatonin on LPS-stimulated bovine mammary epithelial cells.

    Directory of Open Access Journals (Sweden)

    Guang-Min Yu

    Full Text Available Mastitis is the most prevalent disease in dairy cattle worldwide and not only causes huge economic losses in the dairy industry but also threatens public health. To evaluate the therapeutic potential of melatonin in mastitis, we examined the ability of melatonin to protect bovine mammary epithelial cells (bMECs from the harmful effects of lipopolysaccharide (LPS. We found that melatonin inhibited the LPS-binding protein-CD14-TLR4 signaling pathway in bMECs, which had opposing effects on pro-inflammatory and anti-inflammatory mediators. Melatonin decreased LPS-induced expression of pro-inflammatory cytokines, chemokines, and positive acute-phase proteins (APPs, including tumor necrosis factor-α, interleukin (IL-1β, IL-6, granulocyte-monocyte colony-stimulating factor, chemokine CC motif ligand (CCL2, CCL5, serum amyloid A, haptoglobin, C-reactive protein, ceruloplasmin, and α-1 antitrypsin, and increased expression of the anti-inflammatory cytokine IL-1Ra and the negative APP fibrinogen. In addition, melatonin increased dityrosine levels but suppressed nitrite levels by upregulating the expression of Nrf2 and heme oxygenase-1 in the Nrf2 antioxidant defense pathway. Finally, melatonin administration increased the viability of LPS-stimulated bMECs. These results suggest that melatonin protects bMECs from LPS-induced inflammatory and oxidant stress damage and provide evidence that melatonin might have therapeutic utility in mastitis.

  1. The anti-inflammatory and antioxidant effects of melatonin on LPS-stimulated bovine mammary epithelial cells.

    Science.gov (United States)

    Yu, Guang-Min; Kubota, Hirokazu; Okita, Miki; Maeda, Teruo

    2017-01-01

    Mastitis is the most prevalent disease in dairy cattle worldwide and not only causes huge economic losses in the dairy industry but also threatens public health. To evaluate the therapeutic potential of melatonin in mastitis, we examined the ability of melatonin to protect bovine mammary epithelial cells (bMECs) from the harmful effects of lipopolysaccharide (LPS). We found that melatonin inhibited the LPS-binding protein-CD14-TLR4 signaling pathway in bMECs, which had opposing effects on pro-inflammatory and anti-inflammatory mediators. Melatonin decreased LPS-induced expression of pro-inflammatory cytokines, chemokines, and positive acute-phase proteins (APPs), including tumor necrosis factor-α, interleukin (IL)-1β, IL-6, granulocyte-monocyte colony-stimulating factor, chemokine CC motif ligand (CCL)2, CCL5, serum amyloid A, haptoglobin, C-reactive protein, ceruloplasmin, and α-1 antitrypsin, and increased expression of the anti-inflammatory cytokine IL-1Ra and the negative APP fibrinogen. In addition, melatonin increased dityrosine levels but suppressed nitrite levels by upregulating the expression of Nrf2 and heme oxygenase-1 in the Nrf2 antioxidant defense pathway. Finally, melatonin administration increased the viability of LPS-stimulated bMECs. These results suggest that melatonin protects bMECs from LPS-induced inflammatory and oxidant stress damage and provide evidence that melatonin might have therapeutic utility in mastitis.

  2. Personal endotoxin exposure in a panel study of school children with asthma

    Directory of Open Access Journals (Sweden)

    Tjoa Thomas

    2011-08-01

    Full Text Available Abstract Background Endotoxin exposure has been associated with asthma exacerbations and increased asthma prevalence. However, there is little data regarding personal exposure to endotoxin in children at risk, or the relation of personal endotoxin exposure to residential or ambient airborne endotoxin. The relation between personal endotoxin and personal air pollution exposures is also unknown. Methods We characterized personal endotoxin exposures in 45 school children with asthma ages 9-18 years using 376 repeated measurements from a PM2.5 active personal exposure monitor. We also assayed endotoxin in PM2.5 samples collected from ambient regional sites (N = 97 days and from a subset of 12 indoor and outdoor subject home sites (N = 109 and 111 days, respectively in Riverside and Whittier, California. Endotoxin was measured using the Limulus Amoebocyte Lysate kinetic chromogenic assay. At the same time, we measured personal, home and ambient exposure to PM2.5 mass, elemental carbon (EC, and organic carbon (OC. To assess exposure relations we used both rank correlations and mixed linear regression models, adjusted for personal temperature and relative humidity. Results We found small positive correlations of personal endotoxin with personal PM2.5 EC and OC, but not personal PM2.5 mass or stationary site air pollutant measurements. Outdoor home, indoor home and ambient endotoxin were moderately to strongly correlated with each other. However, in mixed models, personal endotoxin was not associated with indoor home or outdoor home endotoxin, but was associated with ambient endotoxin. Dog and cat ownership were significantly associated with increased personal but not indoor endotoxin. Conclusions Daily fixed site measurements of endotoxin in the home environment may not predict daily personal exposure, although a larger sample size may be needed to assess this. This conclusion is relevant to short-term exposures involved in the acute exacerbation of

  3. Metallothionein in mice reduces intestinal zinc loss during acute endotoxin inflammation, but not during starvation or dietary zinc restriction.

    Science.gov (United States)

    Philcox, J C; Sturkenboom, M; Coyle, P; Rofe, A M

    2000-08-01

    Normal metallothionein [(MT)+/+] and MT-null (MT-/-) mice were used to examine the influence of MT on Zn retention and the metabolic consequences of 2 d food deprivation, with and without inflammation induced by intraperitoneal injection of bacterial endotoxin lipopolysaccharide (LPS). LPS reduced fecal Zn concentration in MT+/+ mice from 5.9 +/- 0.2 micromol/g on d 1 to 2.2 +/- 0.2 micromol/g on d 2, but not in MT-/- mice, 5.9 +/- 0.2 and 5.7 +/- 0. 5 micromol/g, respectively. MT+/+ mice fed an 8 mg Zn/kg diet and injected with LPS excreted 40% less Zn over 2 d than their MT-/- counterparts. Starvation for 2 d did not lower fecal Zn concentration in either genotype, although in MT+/+ mice, urinary Zn excretion was reduced from 12.7 +/- 1.3 nmol on d 1 to 5.9 +/- 1.8 nmol on d 2 and plasma Zn concentration was lowered to 9.8 +/- 0.4 micromol/L. Zn was not reduced in urine or plasma of MT-/- mice, with respective values of 10.8 +/- 2.0 nmol on d 1, 9.3 +/- 2.9 nmol on d 2 and 13.0 +/- 1.0 micromol/L. LPS injection resulted in much higher total liver Zn (677 +/- 27 nmol) and MT (106 +/- 2 nmol Cd bound/g) than starvation (Zn = 405 +/- 21, MT = 9 +/- 3) in MT+/+ mice after 2 d, but did not further reduce urinary Zn. LPS-injected MT-/- mice had no rise in liver Zn or fall in plasma and urine Zn. MT-/- mice fed a Zn-deficient (0.8 mg Zn/kg) diet lost 10% of body weight over 25 d compared with no loss in MT+/+ mice. Despite this, MT-/- mice excreted no more Zn via the gut than did MT+/+ mice. In summary, MT inhibits intestinal Zn loss when highly expressed. When uninduced, typically during Zn deficiency, MT appears to conserve Zn and body mass by reducing only urinary and other nonintestinal Zn losses.

  4. Continuous blood purification treatment for endotoxin-induced acute respiratory distress syndrome

    Directory of Open Access Journals (Sweden)

    Y. Jiang

    Full Text Available This study aimed to explore the effects of continuous blood purification (CBP treatment in pigs affected with acute respiratory distress syndrome (ARDS. A total of 12 healthy male pigs, weighing 12±1.8 kg, were randomly and equally assigned to the control and experimental groups. The ARDS pig model was prepared by intravenous injections of endotoxin (20 µg/kg. The control group was given conventional supportive therapy, while the experimental group was given continuous veno-venous hemofiltration therapy. During the treatment process, the variations in dynamic lung compliance, oxygenation index, hemodynamics, and urine volume per hour at different times (Baseline, 0, 2, 4, and 6 h were recorded. The levels of tumor necrosis factor (TNF-α, interleukin 6 (IL-6, and IL-10 in serum and bronchoalveolar lavage fluid (BALF were measured using the enzyme-linked immunosorbent assay. The histomorphological changes of the lung, heart, and kidney were visualized using a light microscope. The nuclear factor κB p65 protein content of the heart, lung, and kidney tissues was also detected using western blot. The experimental group outperformed the control group in both respiratory and hemodynamic events. CBP treatment cleared TNF-α, IL-6, and IL-10 partially from serum and BALF. The pathological examination of the heart, lung, and kidney tissues revealed that the injury was less severe in the experimental group. CBP treatment can improve the organ functions of pigs affected with endotoxin-induced ARDS and protect these organs to some extent.

  5. Role of LPS-elicited signaling in triggering gastric mucosal inflammatory responses to H. pylori: modulatory effect of ghrelin.

    Science.gov (United States)

    Slomiany, B L; Slomiany, A

    2017-08-01

    Infection with Helicobacter pylori is a primary culprit in the etiology of gastric disease, and its cell-wall lipopolysaccharide (LPS) is recognized as a potent endotoxin responsible for triggering a pattern of the mucosal inflammatory responses. The engagement by the LPS of gastric mucosal Toll-like receptor 4 (TLR4) leads to initiation of signal transduction events characterized by the activation of mitogen-activated protein kinase (MAPK) cascade, induction of phosphoinositide-specific phospholipase C (PLC)/protein kinase C (PKC)/phosphatidylinositol 3-kinase (PI3K) pathway, and up-regulation in Src/Akt. These signaling events in turn exert their influence over H. pylori-elicited excessive generation of NO and PGE2 caused by the disturbances in nitric oxide synthase and cyclooxygenase isozyme systems, increase in epidermal growth factor receptor transactivation, and the induction in matrix metalloproteinase-9 (MMP-9) release. Interestingly, the extent of gastric mucosal inflammatory response to H. pylori is influenced by a peptide hormone, ghrelin, the action of which relays on the growth hormone secretagogue receptor type 1a (GHS-R1a)-mediated mobilization of G-protein dependent transduction pathways. Yet, the signals triggered by TLR-4 activation as well as those arising through GHS-R1a stimulation converge at MAPK and PLC/PKC/PI3K pathways that form a key integration node for proinflammatory signals generated by H. pylori LPS as well as for those involved in modulation of inflammation by ghrelin. Hence, therapeutic targeting these signals' convergence and integration node could provide a novel and attractive opportunities for developing more effective treatments of H. pylori-related gastric disease.

  6. THE ROLE OF ENDOTOXIN IN DEVELOPMENT OF SUPPURATIVE-SEPTIC DISEASES AND METHODS OF ENDOTOXIN LEVEL DETERMINATION IN BLOOD

    Directory of Open Access Journals (Sweden)

    E. V. Rusanova

    2013-01-01

    Full Text Available The severity and duration of suppurative inflammation of different locations is directly dependent on the level of lipopolysaccharides (endotoxin in blood. Detection of this value and its dynamics during therapy enables determination of the treatment tactics and its correction. Examination of endotoxin level in biological fluids is a necessary diagnostic component in the therapeutic-and-prophylactic institutions. Today, highly sensitive methods such as LAL-test in different modifications and MAP-test (method of activated particles are widely practised.

  7. Mechanism for Prenatal LPS-Induced DA Neuron Loss

    Science.gov (United States)

    2005-03-01

    under way. Despite these problems, we feel we have made excellent progress. Sol LPS Speciric progress: 1. Prenatal LPS exposure reduces BrdU positive...W.G., Chen, I., Bachus, S.E., Lewitt , P.A., 2003. plasma TNF-alpha levels following intracerebroventricular injection of In situ hybridization for

  8. [Review on characteristics and detecting assay of bacterial endotoxin contamination in water environment].

    Science.gov (United States)

    Zhang, Can; Liu, Wen-Jun; Zhang, Ming-Lu; Tian, Fang; Yang, Yi; An, Dai-Zhi

    2014-04-01

    Endotoxins, also known as lipopolysaccharide complexes, are anchored in the outer membrane cell wall of most Gram-negative bacteria and some cyanobacteria. They are continuously released to environment during cell decay. Being common pyrogens and highly immunogenic molecules, endotoxins are related to many human diseases. Due to the tolerances and thermo-stability of endotoxin molecules, they were hard to be removed by common methods. The health risk caused by the endotoxin contamination in drinking water and water environment by various exposure pathways have attracted more and more attention in recent years. In this paper, the physical and chemical properties, biological activities and detection assay of the endotoxin contamination were reviewed, and interfere factors of the main assay, the LAL/TAL (Limulus amebocyte lysate/Tachypleus amebocyte lysate) assay, for detecting endotoxin in water sample were investigated, and the development tendency of the endotoxin detection assay was analyzed.

  9. Quantification of endotoxins in infected root canals and acute apical abscess exudates: monitoring the effectiveness of root canal procedures in the reduction of endotoxins.

    Science.gov (United States)

    Sousa, Ezilmara L R; Martinho, Frederico C; Nascimento, Gustavo G; Leite, Fabio R M; Gomes, Brenda P F A

    2014-02-01

    This clinical study was conducted to measure the endotoxin levels in infected root canals (RCs) and exudates related to acute apical abscesses (AAAs). In addition, the effectiveness of RC procedures in reducing the endotoxin levels in RCs was monitored. Paired samples of infected RCs and exudates from AAAs were collected from 10 subjects by using paper points. RCs samples were collected before (RCS1) and after chemomechanical preparation (CMP) (RCS2), after 17% EDTA (RCS3), and after 30 days of intracanal medication (Ca[OH]2 + chlorhexidine) (RCS4). A turbidimetric kinetic limulus amebocyte lysate assay was used for the measurement of endotoxins. Endotoxins were detected in 100% of the baseline samples of AAAs and RCs (RCS1) with median values of 175 EU/mL and 41.5 EU/mL, respectively (P endotoxins were reduced to a median value of 0.54 EU/mL (P endotoxin levels (median= 0.37 EU/mL) (P = .07). However, intracanal medication for 30 days (RCS4) reduced endotoxins to median values of 0.03 EU/mL (P endotoxins found in AAAs and RCs collected from the same tooth. Moreover, the effectiveness of CMP in reducing the endotoxin levels from RCs in acute endodontic infection was improved by the use of RC medication. Copyright © 2014 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  10. Plant Polyphenols and Exendin-4 Prevent Hyperactivity and TNF-α Release in LPS-Treated In vitro Neuron/Astrocyte/Microglial Networks

    Directory of Open Access Journals (Sweden)

    Francesca Gullo

    2017-09-01

    Full Text Available Increasing evidence supports a decisive role for neuroinflammation in the neurodegenerative process of several central nervous system (CNS disorders. Microglia are essential mediators of neuroinflammation and can regulate a broad spectrum of cellular responses by releasing reactive oxygen intermediates, nitric oxide, proteases, excitatory amino acids, and cytokines. We have recently shown that also in ex-vivo cortical networks of neurons, astrocytes and microglia, an increased level of tumor necrosis factor-alpha (TNF-α was detected a few hours after exposure to the bacterial endotoxin lipopolysaccharide (LPS. Simultaneously, an atypical “seizure-like” neuronal network activity was recorded by multi-electrode array (MEA electrophysiology. These effects were prevented by minocycline, an established anti-inflammatory antibiotic. We show here that the same inhibitory effect against LPS-induced neuroinflammation is exerted also by natural plant compounds, polyphenols, such as curcumin (CU, curcuma longa, crocin (CR, saffron, and resveratrol (RE, grape, as well as by the glucagon like peptide-1 receptor (GLP-1R agonist exendin-4 (EX-4. The drugs tested also caused per-se early transient (variable changes of network activity. Since it has been reported that LPS-induced neuroinflammation causes rearrangements of glutamate transporters in astrocytes and microglia, we suggest that neural activity could be putatively increased by an imbalance of glial glutamate transporter activity, leading to prolonged synaptic glutamatergic dysregulation.

  11. Effect of Bacterial Endotoxins on Superovulated Mouse Embryos In Vivo: Is CSF-1 Involved in Endotoxin-Induced Pregnancy Loss?

    Directory of Open Access Journals (Sweden)

    Yogesh Kumar Jaiswal

    2006-01-01

    Full Text Available Mammalian embryonic development is regulated by several cytokines and growth factors from embryonic or maternal origins. Since CSF-1 plays important role in embryonic development and implantation, we investigated its role in gram-negative bacterial LPS-induced implantation failure. The effect of LPS on normal (nonsuperovulated and superovulated in vivo-produced embryos was assessed by signs of morphological degeneration. A significantly similar number of morphologically degenerated embryos recovered from both nonsuperovulated and superovulated LPS treated animals on day 2.5 of pregnancy onwards were morphologically and developmentally abnormal as compared to their respective controls (P < .001. Normal CSF-1 expression level and pattern were also altered through the preimplantation period in the mouse embryos and uterine horns after LPS treatment. This deviation from the normal pattern and level of CSF-1 expression in the preimplantation embryos and uterine tissues suggest a role for CSF-1 in LPS-induced implantation failure.

  12. Streptomycetes in house dust: associations with housing characteristics and endotoxin

    Science.gov (United States)

    In addition to mold, indoor bioaerosols also contain bacterial components that may have implications for human health. Endotoxin is a cell wall component in Gram-negative bacteria present at varying levels indoors that has been found to have respiratory health implications. Stre...

  13. Effect of nitric oxide scavengers, carboxy-PTIO on endotoxin ...

    African Journals Online (AJOL)

    Research evidence shows that sepsis-associated vascular relaxation is mediated by nitric oxide. Nitric oxide formation is stimulated by endotoxin, cytokines such as Tumor necrosis factor, and Interleukines. The stimulation is due to the activation of an inducible nitric oxide synthase, which transforms an amino acid ...

  14. Dephosphorylation of endotoxin by alkaline phosphatase in vivo

    NARCIS (Netherlands)

    Poelstra, Klaas; Bakker, W.W; Klok, P.A; Kamps, J.AAM; Hardonk, M.J; Meijer, D.K F

    1997-01-01

    Natural substrates for alkaline phosphatase (AP) are at present not identified despite extensive investigations. Difficulties in imagining a possible physiological function involve its extremely high pH optimum for the usual exogenous substrates and its localization as an ecto-enzyme. As endotoxin

  15. Sporulation and delta-endotoxin synthesis by Bacillus thuringiensis.

    Science.gov (United States)

    Aronson, A

    2002-03-01

    Bacillus thuringiensis is distinguished from the very closely related Bacillus cereus and Bacillus anthracis by the presence of several plasmid-encoded delta-endotoxin genes. These delta-endotoxins, synthesized as protoxins, are produced in large quantities during sporulation and are packaged into intracellular inclusions. Ingestion of the inclusions by insect larvae leads to protoxin solubilization and conversion to toxins each specific for one of several orders of insects. The toxins form cation-selective channels in the membrane of cells lining the larval midgut with subsequent lethality. In most cases, delta-endotoxin synthesis and sporulation are closely coupled. The latter process in B. thuringiensis is probably virtually identical to that in Bacillus subtilis with the additional use of mother cell sporulation forms of RNA polymerase for the synthesis of the delta-endotoxins. There are other more subtle plasmid-encoded functions or plasmid interactions related to regulating protoxin synthesis. Consideration of both plasmid and chromosomal genes is thus critical for defining this organism.

  16. Role of vasoactive amines in the antitumor activity of endotoxin

    NARCIS (Netherlands)

    Hofhuis, F.M.A.; Bloksma, N.; Kuper, C.F.; Willers, J.M.N.

    1984-01-01

    To estimate a possible role of vasoamines in the antitumor action of endotoxin, effects of isoproterenol, serotonin and adrenaline on subcutaneosly transplanted murine Meth A sarcoma and the capacity of these agents to elicit antitumor factors were studied. Macroscopically all agents induced tumor

  17. PRESENCE OR ABSENCE OF ENDOTOXIN AND ANTI-TNF-oz

    African Journals Online (AJOL)

    Short communication. TEST OF ENDOTHELIAL MONOLAYER INTEGRITY IN THE. PRESENCE OR ABSENCE OF ENDOTOXIN AND ANTI-TNF-oz. MONOCLONAL ... vessels using digestive enzymes such as collagen and tripsin, and grow them in culture flasks ... permeability for macromolecules such as plasma proteins.

  18. Optimizing the extraction, storage, and analysis of airborne endotoxins

    Science.gov (United States)

    While the Limulus amebocyte lysate (LAL) assay is part of most procedures to assess airborne endotoxin exposure, there is no universally agreed upon standard procedure. The purpose of this study was to fill in additional knowledge gaps with respect to the extraction, storage, and analysis of endotox...

  19. Endotoxin exposure and atopic sensitization in adult pig farmers

    NARCIS (Netherlands)

    Portengen, L.; Preller, L.; Tielen, M.; Doekes, G.; Heederik, D.

    2005-01-01

    Background: Recent studies have reported a low prevalence of atopic sensitization and respiratory allergy in children growing up on farms. Objectives: We sought to evaluate the dose-response relationship between endotoxin and atopic sensitization in adult farmers and to assess the effect on

  20. A selective sorbent for removing bacterial endotoxins from blood

    Science.gov (United States)

    Morozov, A. S.; Kopitsyna, M. N.; Bessonov, I. V.; Karelina, N. V.; Nuzhdina, A. V.; Sarkisov, I. Yu.; Pavlova, L. A.; Tsyurupa, M. P.; Blinnikova, Z. K.; Davankov, V. A.

    2016-12-01

    Synthetic ligands carrying a positive charge and capable of selective binding of bacterial endotoxins are covalently immobilized on surfaces of domestic hemosorbent Styrosorb-514 based on hypercrosslinked polystyrene. It is shown that the resulting sorbent aimed at treating sepsis exceeds imported specific hemosorbent in Toraymyxin™ columns in removing lipopolysaccharides, and can be used in domestically-produced Desepta columns.

  1. [The importance of endotoxin producing bacterias for practical purposes

    Science.gov (United States)

    Schimmel, Dietrich

    1994-01-01

    Lipopolysaccharides (endotoxin) cause according to resorption out of the intestinal tract or aerogenic inhalation or by a septic infection clinical signs. The clinical reactions are praeshock symptoms, acute forms of shock and death. The experimental intratracheally administration of lipopolysaccharides into calves caused pneumonic lesions without bacterial experimental infection.

  2. Immunological studies on albino rats against crude endotoxins of ...

    African Journals Online (AJOL)

    The genus Aeromonas comprises a group of organisms widely distributed in the environment, isolated from water, shallow tube wells, dug wells, piped water supply from sewage, children and adults with diarrhoea, and associated with disease in animals and man. The present work studied the effect of crude endotoxin ...

  3. Human lipoproteins have divergent neutralizing effects on E. coli LPS, N. meningitidis LPS, and complete Gram-negative bacteria.

    NARCIS (Netherlands)

    Sprong, T.; Netea, M.G.; Ley, P. van der; Verver-Jansen, T.J.G.; Jacobs, L.E.; Stalenhoef, A.F.H.; Meer, J.W.M. van der; Deuren, M. van

    2004-01-01

    The use of lipoproteins has been suggested as a treatment for Gram-negative sepsis because they inhibit lipopolysaccharide (LPS)-mediated cytokine production. However, little is known about the neutralizing effects of lipoproteins on cytokine production by meningococcal LPS or whole Gram-negative

  4. The Deep-Sea Polyextremophile Halobacteroides lacunaris TB21 Rough-Type LPS: Structure and Inhibitory Activity towards Toxic LPS

    Science.gov (United States)

    Di Lorenzo, Flaviana; Palmigiano, Angelo; Paciello, Ida; Pallach, Mateusz; Garozzo, Domenico; Bernardini, Maria-Lina; La Cono, Violetta; Yakimov, Michail M.; Molinaro, Antonio; Silipo, Alba

    2017-01-01

    The structural characterization of the lipopolysaccharide (LPS) from extremophiles has important implications in several biomedical and therapeutic applications. The polyextremophile Gram-negative bacterium Halobacteroides lacunaris TB21, isolated from one of the most extreme habitats on our planet, the deep-sea hypersaline anoxic basin Thetis, represents a fascinating microorganism to investigate in terms of its LPS component. Here we report the elucidation of the full structure of the R-type LPS isolated from H. lacunaris TB21 that was attained through a multi-technique approach comprising chemical analyses, NMR spectroscopy, and Matrix-Assisted Laser Desorption Ionization (MALDI) mass spectrometry. Furthermore, cellular immunology studies were executed on the pure R-LPS revealing a very interesting effect on human innate immunity as an inhibitor of the toxic Escherichia coli LPS. PMID:28653982

  5. Research on Protective Effect and Mechanism of Idazoxan on lps Attacked Acute Hepatic Injury

    Science.gov (United States)

    Zhu, Junyu; Ying, Shangqi; Kang, Wenyuan; Huang, Wenjuan; Liang, Huaping

    2018-01-01

    Objective: To observe the protection effect of Idazoxan (IDA) on LPS induced acute hepatic injury, and to explore its action mechanism. Methods: 60 adult C57BL/6 mice were divided into a control group (20 mice, intraperitoneal injection of phosphate buffer), a model group (20 mice, intraperitoneal injection of LPS 10 mg/kg) and a agmatine group (20 mice, intraperitoneal injection of LPS 10 mg/kg and agmatine 200 mg/kg) according to random number table method. Blood and liver tissue were collected for preparation of tissue homogenate. Enzyme-linked immunosorbent assay (ELISA) was adopted for detecting tumor necrosis factor-α (TNF-α) and interleukin (IL- 1β and IL - 6) contents in the serum and liver tissue at 24h after molding. Automatic biochemical analyzer is used for determining alanine transaminase (ALT), aspartate aminotransferase (AST) and lactate dehydrogenase (LDH) level at 24h after molding; The liver tissue pathology changes were observed at 24h after molding. Macrophage RAW264.7 cells were stimulated by 10 μg/mL LPS and with or without IDA (100 μmol/L). 2’, 7’-dichlorofluoresce in diacetate (DCFH-DA) was used as a fluorescent probe for detection of intracellular reactive oxygen species (ROS) level; qRT - PCR method was used for detecting antioxidant enzymes HO-1 and NQO-1 mRNA expression level at 2h, 4h and 8 h. Results: mice in the model group suffered from depression, curling and food water forbidding at 6h after molding. Mice in the Idazoxan group have obviously better spirit and activity than that of model group. The serum ALT, AST and LDH level of LPS attacked acute hepatic injury mice can be effectively alleviated after Idazoxan treatment. The expression of proinflammatory factor TNF-α and IL-6 in the liver can be reduced. The liver showed obvious pathological changes at 24 h after injection, such as liver cell swelling, necrosis, congestion, inflammatory cell infiltration, etc.; The liver cell injury was prominently alleviated in IDA

  6. Endotoxin contamination and control in surface water sources and a drinking water treatment plant in Beijing, China.

    Science.gov (United States)

    Can, Zhang; Wenjun, Liu; Wen, Sun; Minglu, Zhang; Lingjia, Qian; Cuiping, Li; Fang, Tian

    2013-07-01

    In this paper, endotoxin contamination was determined in treated water following each unit of a drinking water treatment plant (WTP) in Beijing, China and its source water (SW) from a long water diversion channel (Shijiazhuang-Beijing) originating from four reservoirs in Hebei province, China. The total-endotoxin activities in SW ranged from 21 to 41 EU/ml at five selected cross sections of the diversion channel. The total-endotoxin in raw water of the WTP ranged from 11 to 16 EU/ml due to dilution and pretreatment during water transportation from Tuancheng Lake to the WTP, and finished water of the WTP ranged from 4 to 10 EU/ml, showing a 49% decrease following the full-scale treatment process at the WTP. Compared with the 31% removal of free-endotoxin, the WTP removed up to 71% of bound-endotoxin in raw water. The traditional treatment processes (coagulation, sedimentation and filtration) in the WTP removed substantial amounts of total-endotoxin (up to 63%), while endotoxin activities increased after granular activated carbon (GAC) adsorption and chlorination. The total-endotoxin in the actual water was composed of free-endotoxin and bound-endotoxin (endotoxin aggregates, bacteria-bound endotoxins and particle-attached endotoxins). The endotoxin aggregates, bacteria-bound endotoxins and particle-attached endotoxins co-exist as suspended particles in water, and only the bacteria-bound endotoxins were correlated with bacterial cells suspended in water. The particle distribution of endotoxin aggregates in ultrapure water was also tested and the results showed that the majority (64-89%) of endotoxin aggregates had diameters contamination and control in treated water following each unit of the WTP processes and its SW from reservoirs are discussed and compared with regard to bacterial cell counts and particle characteristics, which were dependent, to a certain extent, on different flow rates and turbulence of the water environments. Copyright © 2013 Elsevier Ltd. All

  7. Airborne endotoxin concentrations at a large open-lot dairy in southern idaho.

    Science.gov (United States)

    Dungan, Robert S; Leytem, April B

    2009-01-01

    Endotoxins are derived from gram-negative bacteria and are a potential respiratory health risk for animals and humans. To determine the potential for endotoxin transport from a large open-lot dairy, total airborne endotoxin concentrations were determined at an upwind location (background) and five downwind locations on three separate days. The downwind locations were situated at of the edge of the lot, 200 and 1390 m downwind from the lot, and downwind from a manure composting area and wastewater holding pond. When the wind was predominantly from the west, the average endotoxin concentration at the upwind location was 24 endotoxin units (EU) m(-3), whereas at the edge of the lot on the downwind side it was 259 EU m(-3). At 200 and 1390 m downwind from the edge of the lot, the average endotoxin concentrations were 168 and 49 EU m(-3), respectively. Average airborne endotoxin concentrations downwind from the composting site (36 EU m(-3)) and wastewater holding pond (89 EU m(-3)) and 1390 m from the edge of the lot were not significantly different from the upwind location. There were no significant correlations between ambient weather data collected and endotoxin concentrations over the experimental period. The downwind data show that the airborne endotoxin concentrations decreased exponentially with distance from the lot edge. Decreasing an individual's proximity to the dairy should lower their risk of airborne endotoxin exposure and associated health effects.

  8. Endotoxin inactivation via steam-heat treatment in dilute simethicone emulsions used in biopharmaceutical processes.

    Science.gov (United States)

    Britt, Keith A; Galvin, Jeffrey; Gammell, Patrick; Nti-Gyabaah, Joseph; Boras, George; Kolwyck, David; Ramirez, José G; Presente, Esther; Naugle, Gregory

    2014-01-01

    Simethicone emulsion is used to regulate foaming in cell culture operations in biopharmaceutical processes. It is also a potential source of endotoxin contamination. The inactivation of endotoxins in dilute simethicone emulsions was assessed as a function of time at different steam temperatures using a Limulus amebocyte lysate kinetic chromogenic technique. Endotoxin inactivation from steam-heat treatment was fit to a four-parameter double exponential decay model, which indicated that endotoxin inactivation was biphasic, consisting of fast and slow regimes. In the fast regime, temperature-related effects were dominant. Transitioning into the slow regime, the observed temperature dependence diminished, and concentration-related effects became increasingly significant. The change in the Gibbs free energy moving through the transition state indicated that a large energy barrier must be overcome for endotoxin inactivation to occur. The corresponding Arrhenius pre-exponential factor was >10(12) s(-1) suggesting that endotoxins in aqueous solution exist as aggregates. The disorder associated with the endotoxin inactivation reaction pathway was assessed via the change in entropy moving through the transition state. This quantity was positive indicating that endotoxin inactivation may result from hydrolysis of individual endotoxin molecules, which perturbs the conformation of endotoxin aggregates, thereby modulating the biological activity observed. Steam-heat treatment decreased endotoxin levels by 1-2 logarithm (log) reduction (LRV), which may be practically relevant depending on incoming raw material endotoxin levels. Antifoam efficiency and cell culture performance were negligibly impacted following steam-heat treatment. The results from this study show that steam-heat treatment is a viable endotoxin control strategy that can be implemented to support large-scale biopharmaceutical manufacturing. © 2014 American Institute of Chemical Engineers.

  9. Innate immune defenses exhibit circadian rhythmicity and differential temporal sensitivity to a bacterial endotoxin in Nile tilapia (Oreochromis niloticus)

    DEFF Research Database (Denmark)

    Lazado, Carlo Cabacang; Skov, Peter Vilhelm; Pedersen, Per Bovbjerg

    2016-01-01

    exposed at ZT15. Taken together, this study shows that several key components of humoral immunity in tilapia exhibit circadian rhythms and adapt to photoperiodic changes. Further, results of the bacterial endotoxin challenge suggest that responsiveness of serum humoral factors to a biological insult......:24D (DD), for 20 days to characterize the rhythms of humoral immunity. Serum alkaline phosphatase (ALP), lysozyme (LYZ), peroxidase (PER) and protease (PRO) exhibited significant rhythmicity under LD but not in DD. No significant rhythms were observed in esterase (ESA) and anti-protease (ANTI......) in both photoperiod conditions. Fish reared under LD were subsequently subjected to DD while the group previously under DD was exposed to LD, and this carried on for 3 days before another set of samples was collected. Results revealed that the rhythms of LYZ, PER and PRO but not ALP persisted when...

  10. [The idiosyncratic hepatotoxicity of Polygonum multiflorum based on endotoxin model].

    Science.gov (United States)

    Li, Chun-yu; Li, Xiao-fei; Tu, Can; Li, Na; Ma, Zhi-jie; Pang, Jing-yao; Jia, Ge-liu-chang; Cui, He-rong; You, Yun; Song, Hai-bo; Du, Xiao-xi; Zhao, Yan-ling; Wang, Jia-bo; Xiao, Xiao-he

    2015-01-01

    The liver injury induced by Polygonum multiflorum Thunb. (PM) was investigated based on idiosyncratic hepatotoxicity model co-treated with lipopolysaccharide (LPS) at a non-hepatotoxic dose. Sprague-Dawley (SD) rats were intragastrically administered with three doses (18.9, 37.8, 75.6 g crude drug per kg body weight) of 50% alcohol extracts of PM alone or co-treated with non-toxic dose of LPS (2.8 mg·kg(-1)) via tail vein injection. The plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities were assayed and the isolated livers were evaluated for histopathological changes. The dose-toxicity relationships of single treatment of PM or co-treatment of LPS were investigated comparatively to elucidate the idiosyncratic hepatotoxicity of PM. The results showed that no significant alterations of plasma ALT and AST activities were observed in the groups of solo-administration of LPS (2.8 mg·kg(-1), i.v.) or different dosage (18.9, 37.8 and 75.6 g·kg(-1), i.g.) of PM, compared to normal control group (P > 0.05); while significant elevations were observed in the co-administration groups of PM and LPS. Treatment with LPS alone caused slight infiltration of inflammatory cells in portal area but no evident hepatocytes injury. Co-treatment with LPS and PM (75.6 g·kg(-1), i.g.) caused hepatocyte focal necrosis, loss of central vein intima and a large number of inflammatory cell infiltration in portal areas. When further reduce the dosage of PM, significant increases of plasma ALT and AST activities (P < 0.05) were still observed in co-administration groups of LPS and PM (1.08 or 2.16 g·kg(-1)), but not in LPS or PM solo-administration groups. Nevertheless, the co-treatment of low dosage of PM (0.54 g·kg(-1)) with LPS did not induce any alteration of plasma ALT and AST. In conclusion, intragastric administration with 75.6 g·kg(-1) of PM did not induce liver injury in normal rats model; while the 2 folds of clinical equivalent dose of PM (1.08 g

  11. Interleukin-23 mediates the pathogenesis of LPS/GalN-induced liver injury in mice.

    Science.gov (United States)

    Bao, Suxia; Zhao, Qiang; Zheng, Jianming; Li, Ning; Huang, Chong; Chen, Mingquan; Cheng, Qi; Zhu, Mengqi; Yu, Kangkang; Liu, Chenghai; Shi, Guangfeng

    2017-05-01

    Interleukin-23 (IL-23) is required for T helper 17 (Th17) cell responses and IL-17 production in hepatitis B virus infection. A previous study showed that the IL-23/IL-17 axis aggravates immune injury in patients with chronic hepatitis B virus infection. However, the role of IL-23 in acute liver injury remains unclear. The purpose of this study was to determine the role of the inflammatory cytokine IL-23 in lipopolysaccharide/d-galactosamine (LPS/GalN)-induced acute liver injury in mice. Serum IL-23 from patients with chronic hepatitis B virus (CHB), acute-on-chronic liver failure (ACLF) and healthy individuals who served as healthy controls (HCs) was measured by ELISA. An IL-23p19 neutralizing antibody or an IL-23p40 neutralizing antibody was administered intravenously at the time of challenge with LPS (10μg/kg) and GalN (400mg/kg) in C57BL/6 mice. Hepatic pathology and the expression of Th17-related cytokines, including IL-17 and TNF-α; neutrophil chemoattractants, including Cxcl1, Cxcl2, Cxcl9, and Cxcl10; and the stabilization factor Csf3 were assessed in liver tissue. Serum IL-23 was significantly upregulated in ACLF patients compared with CHB patients and HCs (Pliver tissue histopathology and significant reductions in the expression of Th17-related inflammatory cytokine, including IL-17 and TNF-α; neutrophil chemoattractants, including Cxcl1, Cxcl2, Cxcl9, and Cxcl10; and stabilization factors Csf3 within the liver tissue compared with LPS/GalN mice (Pliver injury in mice. IL-23 neutralizing antibodies attenuated liver injury by reducing the expression of Th17-related inflammatory cytokines, neutrophil chemoattractants and stabilization factors within the liver tissue, which indicated that IL-23 likely functions upstream of Th17-related cytokine and chemokine expression to recruit inflammatory cells into the liver. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. Endotoxin removal by radio frequency gas plasma (glow discharge)

    Science.gov (United States)

    Poon, Angela

    2011-12-01

    Contaminants remaining on implantable medical devices, even following sterilization, include dangerous fever-causing residues of the outer lipopolysaccharide-rich membranes of Gram-negative bacteria such as the common gut microorganism E. coli. The conventional method for endotoxin removal is by Food & Drug Administration (FDA)-recommended dry-heat depyrogenation at 250°C for at least 45 minutes, an excessively time-consuming high-temperature technique not suitable for low-melting or heat-distortable biomaterials. This investigation evaluated the mechanism by which E. coli endotoxin contamination can be eliminated from surfaces during ambient temperature single 3-minute to cumulative 15-minute exposures to radio-frequency glow discharge (RFGD)-generated residual room air plasmas activated at 0.1-0.2 torr in a 35MHz electrodeless chamber. The main analytical technique for retained pyrogenic bio-activity was the Kinetic Chromogenic Limulus Amebocyte Lysate (LAL) Assay, sufficiently sensitive to document compliance with FDA-required Endotoxin Unit (EU) titers less than 20 EU per medical device by optical detection of enzymatic color development corresponding to LAL Assay of sterile water extracts from both glass and Ge specimens. The Ge prism MAIR-IR measurements were repeated after employing 3-minute RFGD treatments sequentially for more than 10 cycles to observe removal of deposited matter that correlated with diminished EU titers. The results showed that 5 cycles, for a total exposure time of 15 minutes to low-temperature gas plasma, was sufficient to reduce endotoxin titers to below 0.05 EU/ml, and correlated with concurrent reduction of major endotoxin reference standard absorption bands at 3391 cm-1, 2887 cm-1, 1646 cm -1 1342 cm-1, and 1103 cm-1 to less than 0.05 Absorbance Units. Band depletion varied from 15% to 40% per 3-minute cycle of RFGD exposure, based on peak-to-peak analyses. In some cases, 100% of all applied biomass was removed within 5 sequential

  13. Exposure to airborne microorganisms and endotoxin in herb processing plants.

    Science.gov (United States)

    Dutkiewicz, J; Krysińska-Traczyk, E; Skórska, C; Sitkowska, J; Prazmo, Z; Golec, M

    2001-01-01

    Microbiological air sampling was performed in two herb processing plants located in eastern Poland. Air samples for determination of the levels of bacteria, fungi, dust and endotoxin were collected at 14 sites during cleaning, cutting, grinding, sieving, sorting and packing of 11 kinds of herbs (nettle, caraway, birch, celandine, marjoram, mint, peppermint, sage, St. John's wort, calamus, yarrow), used for production of medications, cosmetics and spices. It was found that processing of herbs was associated with a very high pollution of the air with bacteria, fungi, dust and endotoxin. The numbers of microorganisms (bacteria and fungi) in the air of herb processing plants ranged within 40.6-627.4 x 10(3) cfu/m3 (mean +/- S.D = 231.4 +/- 181.0 x 10(3) cfu/m3). The greatest concentrations were noted at the initial stages of production cycle, during cleaning, cutting and grinding of herbs. The numbers of airborne microorganisms were also significantly (pair of herb processing plants were mesophilic bacteria, among which endospore-forming bacilli (Bacillus spp.) and actinomycetes of the species Streptomyces albus were most numerous. Among Gram-negative bacteria, the most common was endotoxin-producing species Alcaligenes faecalis. Altogether, 37 species or genera of bacteria and 23 species or genera of fungi were identified in the air of herb processing plants, of these, 11 and 10 species or genera respectively were reported as having allergenic and/or immunotoxic properties. The concentrations of dust and bacterial endotoxin in the air of herb processing plants were large with extremely high levels at some sampling sites. The concentrations of airborne dust ranged within 3.2-946.0 mg/m3 (median 18.1 mg/m3), exceeding at 13 out of 14 sampling sites the Polish OEL value of 4 mg/m3. The concentrations of airborne endotoxin ranged within 0.2-2681.0 microg/m3 (median 16.0 microg/m3), exceeding at all sampling sites the suggested OEL value of 0.1 microg/m3. In conclusion

  14. Potentiation of LPS-Induced Apoptotic Cell Death in Human Hepatoma HepG2 Cells by Aspirin via ROS and Mitochondrial Dysfunction: Protection by N-Acetyl Cysteine.

    Directory of Open Access Journals (Sweden)

    Haider Raza

    Full Text Available Cytotoxicity and inflammation-associated toxic responses have been observed to be induced by bacterial lipopolysaccharides (LPS in vitro and in vivo respectively. Use of nonsteroidal anti-inflammatory drugs (NSAIDs, such as aspirin, has been reported to be beneficial in inflammation-associated diseases like cancer, diabetes and cardiovascular disorders. Their precise molecular mechanisms, however, are not clearly understood. Our previous studies on aspirin treated HepG2 cells strongly suggest cell cycle arrest and induction of apoptosis associated with mitochondrial dysfunction. In the present study, we have further demonstrated that HepG2 cells treated with LPS alone or in combination with aspirin induces subcellular toxic responses which are accompanied by increase in reactive oxygen species (ROS production, oxidative stress, mitochondrial respiratory dysfunction and apoptosis. The LPS/Aspirin induced toxicity was attenuated by pre-treatment of cells with N-acetyl cysteine (NAC. Alterations in oxidative stress and glutathione-dependent redox-homeostasis were more pronounced in mitochondria compared to extra- mitochondrial cellular compartments. Pre-treatment of HepG2 cells with NAC exhibited a selective protection in redox homeostasis and mitochondrial dysfunction. Our results suggest that the altered redox metabolism, oxidative stress and mitochondrial function in HepG2 cells play a critical role in LPS/aspirin-induced cytotoxicity. These results may help in better understanding the pharmacological, toxicological and therapeutic properties of NSAIDs in cancer cells exposed to bacterial endotoxins.

  15. Citrulline decreases hepatic endotoxin-induced injury in fructose-induced non-alcoholic liver disease: an ex vivo study in the isolated perfused rat liver.

    Science.gov (United States)

    Ouelaa, Wassila; Jegatheesan, Prasanthi; M'bouyou-Boungou, Japhète; Vicente, Christelle; Nakib, Samir; Nubret, Esther; De Bandt, Jean-Pascal

    2017-06-01

    Steatosis can sensitise the liver to various challenges and favour the development of non-alcoholic fatty liver disease (NAFLD). In this context, fructose feeding promotes endotoxin translocation from the gut, contributing to disease progression via an inflammatory process. Citrulline is protective against fructose-induced NAFLD; we hypothesised that this property might be related to its anti-inflammatory and antioxidative action against endotoxin-induced hepatic injuries. This hypothesis was evaluated in a model of perfused liver isolated from NAFLD rats. Male Sprague-Dawley rats (n 30) were fed either a standard rodent chow or a 60 % fructose diet alone, or supplemented with citrulline (1 g/kg per d) for 4 weeks. After an evaluation of their metabolic status, fasted rats received an intraperitoneal injection of lipopolysaccharide (LPS) (2·5 mg/kg). After 1 h, the livers were isolated and perfused for 1 h to study liver function and metabolism, inflammation and oxidative status. In vivo, citrulline significantly decreased dyslipidaemia induced by a high-fructose diet and insulin resistance. In the isolated perfused rat livers, endotoxaemia resulted in higher cytolysis (alanine aminotransferase release) and higher inflammation (Toll-like receptor 4) in livers of fructose-fed rats, and it was prevented by citrulline supplementation. Oxidative stress and antioxidative defences were similar in all three groups. Amino acid exchanges and metabolism (ammonia and urea release) were only slightly different between the three groups. In this context of mild steatosis, our results suggest that fructose-induced NAFLD leads to an increased hepatic sensitivity to LPS-induced inflammation. Citrulline-induced restriction of the inflammatory process may thus contribute to the prevention of NAFLD.

  16. Gelsolin Inhibits the Inflammatory Process Induced by LPS

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    Ying Cheng

    2017-01-01

    Full Text Available Background/Aims: Endotoxemia is a life-threatening situation that signifies a key challenge in the field of intensive care medicine. Proinflammatory mediators produced by macrophages play a key role in endotoxemia. Gelsolin (GSN is involved in the process of inflammation. Methods: IL-6 and TNF-α in the supernatant were measured with an ELISA kit. NO production was assessed by measurement of nitrite concentration with the Griess assay. si-RNA directed against GSN (si-GSN was transfected by Lipofectamine. Results: LPS decreased the levels of GSN. Recombinant GSN inhibited the cytokines induced by LPS and rescued mice from LPS-induced death, and si-GSN increased death in the LPS-pretreated mice. Conclusion: GSN exhibited a protective role in endotoxemia.

  17. Endotoxin Contamination in Nanomaterials Leads to the Misinterpretation of Immunosafety Results

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    Yang Li

    2017-05-01

    Full Text Available Given the presence of engineered nanomaterials in consumers’ products and their application in nanomedicine, nanosafety assessment is becoming increasingly important. In particular, immunosafety aspects are being actively investigated. In nanomaterial immunosafety testing strategies, it is important to consider that nanomaterials and nanoparticles are very easy to become contaminated with endotoxin, which is a widespread contaminant coming from the Gram-negative bacterial cell membrane. Because of the potent inflammatory activity of endotoxin, contaminated nanomaterials can show inflammatory/toxic effects due to endotoxin, which may mask or misidentify the real biological effects (or lack thereof of nanomaterials. Therefore, before running immunosafety assays, either in vitro or in vivo, the presence of endotoxin in nanomaterials must be evaluated. This calls for using appropriate assays with proper controls, because many nanomaterials interfere at various levels with the commercially available endotoxin detection methods. This also underlines the need to develop robust and bespoke strategies for endotoxin evaluation in nanomaterials.

  18. Alliin, a Garlic (Allium sativum Compound, Prevents LPS-Induced Inflammation in 3T3-L1 Adipocytes

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    Saray Quintero-Fabián

    2013-01-01

    Full Text Available Garlic (Allium sativum L. has been used to alleviate a variety of health problems due to its high content of organosulfur compounds and antioxidant activity. The main active component is alliin (S-allyl cysteine sulfoxide, a potent antioxidant with cardioprotective and neuroprotective actions. In addition, it helps to decrease serum levels of glucose, insulin, triglycerides, and uric acid, as well as insulin resistance, and reduces cytokine levels. However its potential anti-inflammatory effect is unknown. We examined the effects of alliin in lipopolysaccharide- (LPS- stimulated 3T3-L1 adipocytes by RT-PCR, Western blot, and microarrays analysis of 22,000 genes. Incubation of cells for 24 h with 100 μmol/L alliin prevented the increase in the expression of proinflammatory genes, IL-6, MCP-1, and Egr-1 in 3T3-L1 adipocytes exposed to 100 ng/mL LPS for 1 h. Interestingly, the phosphorylation of ERK1/2, which is involved in LPS-induced inflammation in adipocytes, was decreased following alliin treatment. Furthermore, the gene expression profile by microarrays evidentiate an upregulation of genes involved in immune response and downregulation of genes related with cancer. The present results have shown that alliin is able to suppress the LPS inflammatory signals by generating an anti-inflammatory gene expression profile and by modifying adipocyte metabolic profile.

  19. Riluzole partially rescues age-associated, but not LPS-induced, loss of glutamate transporters and spatial memory.

    Science.gov (United States)

    Brothers, Holly M; Bardou, Isabelle; Hopp, Sarah C; Kaercher, Roxanne M; Corona, Angela W; Fenn, Ashley M; Godbout, Jonathan P; Wenk, Gary L

    2013-12-01

    Impaired memory may result from synaptic glutamatergic dysregulation related to chronic neuroinflammation. GLT1 is the primary excitatory amino acid transporter responsible for regulating extracellular glutamate levels in the hippocampus. We tested the hypothesis that if impaired spatial memory results from increased extracellular glutamate due to age or experimentally induced chronic neuroinflammation in the hippocampus, then pharmacological augmentation of the glutamate transporter GLT1 will attenuate deficits in a hippocampal-dependent spatial memory task. The profile of inflammation-related genes and proteins associated with normal aging, or chronic neuroinflammation experimentally-induced via a four-week LPS infusion into the IV(th) ventricle, were correlated with performance in the Morris water maze following treatment with Riluzole, a drug that can enhance glutamate clearance by increasing GLT1 expression. Age-associated inflammation was qualitatively different from LPS-induced neuro-inflammation in young rats. LPS produced a pro-inflammatory phenotype characterized by increased IL-1ß expression in the hippocampus, whereas aging was not associated with a strong central pro-inflammatory response but with a mixed peripheral immune phenotype. Riluzole attenuated the spatial memory impairment, the elevation of serum cytokines and the decrease in GLT1 gene expression in Aged rats, but had no effect on young rats infused with LPS. Our findings highlight the therapeutic potential of reducing glutamatergic function upon memory impairment in neurodegenerative diseases associated with aging.

  20. High exposure to endotoxin in farming is associated with less new-onset pollen sensitisation

    DEFF Research Database (Denmark)

    Elholm, Grethe; Schlünssen, Vivi; Doekes, Gert

    2017-01-01

    in relation to cumulative endotoxin exposure during follow-up, considering early life farm exposure. RESULTS: Endotoxin exposure during follow-up was significantly associated with less new onset of specifically grass and birch pollen sensitisation. For the highest versus lowest quartile of cumulative...... was not associated with significant changes in cat dander and house dust mite sensitisation. CONCLUSIONS: High exposure to endotoxin during young adulthood appears to protect against new onset of pollen sensitisation, independent of childhood farm exposure....

  1. Sensitisation to common allergens and respiratory symptoms in endotoxin exposed workers: a pooled analysis

    DEFF Research Database (Denmark)

    Basinas, Ioannis; Schlünssen, Vivi; Heederik, Dick

    2012-01-01

    Objective To test the hypotheses that current endotoxin exposure is inversely associated with allergic sensitisation and positively associated with non-allergic respiratory diseases in four occupationally exposed populations using a standardised analytical approach. Methods Data were pooled from....... Heterogeneity was primarily present for biofuel workers. Conclusions Occupational endotoxin exposure has a protective effect on allergic sensitisation and hay fever but increases the risk for organic dust toxic syndrome and chronic bronchitis. Endotoxin's protective effects are most clearly observed among...

  2. Endotoxins in baled cottons and airborne dusts in textile mills in the People's Republic of China.

    Science.gov (United States)

    Olenchock, S A; Christiani, D C; Mull, J C; Ye, T T; Lu, P L

    1983-01-01

    Bulk cotton samples and airborne vertical elutriated cotton dusts were obtained from textile mills in Shanghai, People's Republic of China. Analysis of endotoxin contents revealed that baled cottons which were grown in different countries varied in endotoxin contamination. The two textile mills, which operated at similar overall airborne dust levels, differed markedly in the levels of airborne endotoxins. The data suggest that the biological activity or "toxicity" of airborne cotton dusts may not be correlated directly with gravimetric dust levels. PMID:6639029

  3. Histamine Liberation by Cotton Dust Extracts: Evidence Against its Causation by Bacterial Endotoxins

    Science.gov (United States)

    Antweiler, H.

    1961-01-01

    The recently stated hypothesis that histamine liberation by cotton dust extracts is caused by endotoxins from gram-negative bacteria contained in cotton was studied experimentally in various animals. It was shown clearly that such amounts of endotoxin as were found in cotton dust did not give rise to histamine liberation which was provoked by extracts from cotton card-room dusts. Further arguments are given which testify against the endotoxin hypothesis. PMID:13683810

  4. Bioactive and total endotoxins in atmospheric aerosols in the Pearl River Delta region, China

    Science.gov (United States)

    Cheng, Jessica Y. W.; Hui, Esther L. C.; Lau, Arthur P. S.

    2012-02-01

    Endotoxin, a toxic and pyrogenic substance in gram-negative bacteria in atmospheric aerosols was measured over a period of one year at Nansha, Guangzhou and Hong Kong in the Pearl River Delta region, China. Atmospheric aerosols were collected by high-volume samplers. The bioactive endotoxin levels in the samples were determined using the Limulus Amebocyte Lysate (LAL) assay after extraction with pyrogen-free water while the total endotoxin levels were measured by quantifying the biomarker, 3-hydroxy fatty acids (3-OHFAs) with GC-MS. Results showed that there was no significant difference (0.19 endotoxin level in PM 10 among sites (average concentrations ranged from 0.34 to 0.39 EU m -3). However, Hong Kong showed a significantly lower ( p endotoxin level in PM 10 (average of 17.4 ng m -3) compared with Nansha's 29.4 ng m -3 and Guangzhou's 32.7 ng m -3. The bioactive endotoxins were found to be associated with the coarse mode (PM 2.5-10) of the particulates of natural origins while the total endotoxins were associated more with the fine mode (PM 2.5) of the particulates of anthropogenic origins. When normalized with particulate mass, the endotoxin loading is much higher in summer as a result of the increased growth of the bacteria when climatic conditions are favorable. The chemically determined total endotoxins were 3-4 orders of magnitude higher than the bioactive endotoxins quantified using the LAL assay. Correlation analyses between the bioactive endotoxins and 3-OHFAs with different carbon length were analyzed. Results showed that the correlations detected vary among sites and particulate sizes. Although no generalization between the total and bioactive endotoxins can be drawn from the study, the levels reported in this study suggests that the discrepancies between the two measurement approaches, and the bioactive potential of 3-OHFAs with individual carbon chains deserve further investigation.

  5. Epinephrine Enhances the Response of Macrophages under LPS Stimulation

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    Jianyun Zhou

    2014-01-01

    Full Text Available Trauma associated with infection may directly trigger a neuroendocrine reaction in vivo while the hormone epinephrine is known to mediate immune responses to inflammation after injury. However, the role of epinephrine during the earliest stage of trauma still remains unclear. We therefore explored the role of epinephrine on activated macrophages under LPS stimulation in vitro as well as the mechanisms underlying its effect. Dose- and time-dependent effects of epinephrine on macrophage immune function were assessed after LPS activation. We also employed CD14 siRNA interference to investigate whether CD14 played a role in the mechanism underlying the effect of epinephrine on LPS-induced macrophage responses. Our results showed that epinephrine pretreatment (10 ng/mL significantly promoted immune responses from LPS stimulated macrophages, including phagocytic rate, phagocytic index, TNFα/IL-1β/IL-10 secretion, and CD14 expression (P < 0.05. Moreover, TNFα/IL-1β/IL-10 levels attained their peak value 1 hour after incubation with 10 ng/mL epinephrine (P < 0.05, and CD14 siRNA transfection dramatically decreased phagocytosis and cytokine secretion by LPS-activated macrophages (P < 0.05. We therefore conclude that 10 ng/mL epinephrine enhances immune responses from macrophages under LPS stimulation and that the underlying mechanism may relate to CD14 upregulation on the surface of macrophages.

  6. Immunomodulatory effects of honey cannot be distinguished from endotoxin

    DEFF Research Database (Denmark)

    Timm, Michael; Bartelt, Stine; Hansen, Erik Wind

    2008-01-01

    In recent years, the use of honey has re-emerged as a remedy for wound treatment. Effects of honey have been related to the presence of an unidentified component that induces release of inflammatory cytokines from monocytic cells. The present study was intended to further characterize the reported...... in vitro effects of honey. Our results show that natural honeys induce interleukin-6 release from Mono Mac 6 cells as well as release of reactive oxygen species from all-trans retinoic acid (ATRA) differentiated HL-60 cells. The natural honeys contained substantial amounts of endotoxin, and the responses...... observed in the cell based assays were similar to the responses induced by endotoxin alone. In addition, we determined that the immunomodulatory component present in the natural honeys was retained in the ultra filtrated fraction with a molecular weight greater than 20 kDa. The component was resistant...

  7. Endotoxin-like activity associated with Lyme disease Borrelia.

    Science.gov (United States)

    Fumarola, D; Munno, I; Marcuccio, C; Miragliotta, G

    1986-12-01

    The newly recognized spirochete, Borrelia burgdorferi, the causative agent of Lyme Disease, has been examined for endotoxin-like activities as measured by the standard Farmacopea Ufficiale della Republica Italiana rabbit fever test and the Limulus amoebocyte lysate assay. The suspension of heat-killed microorganism caused a febrile response at a dose of 1 X 10(8) bacteria pro kilo. Similar results were obtained in the Limulus assay where the heat-killed spirochetes stimulated formation of solid clot until the concentration of 1 X 10(5) per ml. Both in pyrogen test and in Limulus assay heat-killed Escherichia coli exhibited a higher degree of potency. These results show that LD-Borrelia possess endotoxin-like activities which could help in understanding the pathogenesis of the clinical symptomatology of the disease.

  8. Occupational exposure levels of bioaerosol components are associated with serum levels of the acute phase protein Serum Amyloid A in greenhouse workers

    DEFF Research Database (Denmark)

    Madsen, Anne Mette; Thilsing, Trine; Bælum, Jesper

    2016-01-01

    to elevated levels of bioaerosols. The objective of this study is to assess whether greenhouse workers personal exposure to bioaerosol components was associated with serum levels of the acute phase proteins Serum Amyloid A (SAA) and C-reactive protein (CRP). METHODS: SAA and CRP levels were determined...... in serum sampled repeatedly from 33 greenhouse workers. Blood was drawn repeatedly on Mondays and Thursdays during work weeks. Acute phase protein levels were compared to levels in a comparison group of 42 people and related to individual exposure levels to endotoxin, dust, bacteria, fungi and β......-glucan. RESULTS: Serum levels of SAA and CRP were not significantly different in greenhouse workers and a reference group, or on the two work days. In a mixed model, SAA levels were positively associated with endotoxin exposure levels (p = 0.0007). Results for fungi were not clear. CRP levels were positively...

  9. Respiratory health effects of exposure to low levels of airborne endotoxin - a systematic review.

    Science.gov (United States)

    Farokhi, Azadèh; Heederik, Dick; Smit, Lidwien A M

    2018-02-08

    Elevated endotoxin levels have been measured in ambient air around livestock farms, which is a cause of concern for neighbouring residents. There is clear evidence that occupational exposure to high concentrations of airborne endotoxin causes respiratory inflammation, respiratory symptoms and lung function decline. However, health effects of exposure to low levels of endotoxin are less well described. The aim of this systematic review is to summarize published associations between exposure to relatively low levels of airborne endotoxin and respiratory health endpoints. Studies investigating respiratory effects of measured or modelled exposure to low levels of airborne endotoxin (average respiratory symptoms and lung function. However, considerable heterogeneity existed in the outcomes of the included studies and no overall estimate could be provided by meta-analysis to quantify the possible relationship. Instead, a best evidence synthesis was performed among studies examining the exposure-response relationship between endotoxin and respiratory outcomes. Significant exposure-response relationships between endotoxin and symptoms and FEV1 were shown in several studies, with no conflicting findings in the studies included in the best evidence synthesis. Significantly different effects of endotoxin exposure were also seen in vulnerable subgroups (atopics and patients with broncho-obstructive disease) and smokers. Respiratory health effects of exposure to low levels of airborne endotoxin (respiratory health effects, especially in vulnerable subgroups of the population.

  10. Changes in serum levels of lipopolysaccharides and CD26 in patients with Crohn's disease

    Science.gov (United States)

    Kotze, Paulo Gustavo; Martinez, Carlos Augusto Real; Camargo, Michel Gardere; Guadagnini, Dioze; Calixto, Antonio Ramos; Vasques, Ana Carolina Junqueira; Ayrizono, Maria de Lourdes Setsuko; Geloneze, Bruno; Pareja, José Carlos; Saad, Mario José; Coy, Claudio Saddy Rodrigues

    2017-01-01

    Background/Aims Lipopolysaccharide (LPS) is a molecule formed by lipids and polysaccharides and is the major cell wall component of gram-negative bacteria. High LPS levels are known to block CD26 expression by activating Toll-like receptor 4. The aim of this study was to correlate the serum levels of LPS and CD26 in Crohn's disease (CD) patients with serum levels of C-reactive protein (CRP), interleukins, CD activity index, and tumor necrosis factor-α (TNF-α). Methods Serum samples were collected from 27 individuals (10 with active CD, 10 with inactive CD, and 7 controls) and the levels of LPS, CD26, TNF-α, interleukin-1β (IL-1β), IL-6, IL-17, and CRP were determined by enzyme-linked immunosorbent assay. The levels of LPS and CD26 were then tested for correlation with TNF-α, IL-1β, IL-6, IL-17, and CRP. Results Serum levels of LPS were significantly elevated in the active CD group (P=0.003). Levels of IL-1β (P=0.002), IL-6 (P=0.003), and IL-17 (P<0.001) were lower in the CD groups. Serum TNF-α levels were increased in the active CD group. The CRP levels were elevated in the CD groups when compared to controls (P<0.001). The CD26 levels were lower in the CD groups than in the control group (P<0.001). Among the variables analyzed, there was a correlation between LPS and CRP (r=−0.53, P=0.016) in the CD groups. Conclusions Individuals with CD exhibited higher serum levels of LPS varying from a 2- to 6-fold increase depending on disease activity, when compared with healthy controls. CD26 levels were lower in the CD groups. Both LPS and CD26 correlated with disease severity and serve as potential CD biomarkers. PMID:28670232

  11. Changes in serum levels of lipopolysaccharides and CD26 in patients with Crohn's disease

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    Daniéla Oliveira Magro

    2017-07-01

    Full Text Available Background/Aims: Lipopolysaccharide (LPS is a molecule formed by lipids and polysaccharides and is the major cell wall component of gram-negative bacteria. High LPS levels are known to block CD26 expression by activating Toll-like receptor 4. The aim of this study was to correlate the serum levels of LPS and CD26 in Crohn's disease (CD patients with serum levels of C-reactive protein (CRP, interleukins, CD activity index, and tumor necrosis factor-α (TNF-α.Methods: Serum samples were collected from 27 individuals (10 with active CD, 10 with inactive CD, and 7 controls and the levels of LPS, CD26, TNF-α, interleukin-1β (IL-1β, IL-6, IL-17, and CRP were determined by enzyme-linked immunosorbent assay. The levels of LPS and CD26 were then tested for correlation with TNF-α, IL-1β, IL-6, IL-17, and CRP.Results: Serum levels of LPS were significantly elevated in the active CD group (P=0.003. Levels of IL-1β (P=0.002, IL-6 (P=0.003, and IL-17 (P<0.001 were lower in the CD groups. Serum TNF-α levels were increased in the active CD group. The CRP levels were elevated in the CD groups when compared to controls (P<0.001. The CD26 levels were lower in the CD groups than in the control group (P<0.001. Among the variables analyzed, there was a correlation between LPS and CRP (r=−0.53, P=0.016 in the CD groups.Conclusions: Individuals with CD exhibited higher serum levels of LPS varying from a 2- to 6-fold increase depending on disease activity, when compared with healthy controls. CD26 levels were lower in the CD groups. Both LPS and CD26 correlated with disease severity and serve as potential CD biomarkers.

  12. Screening extemporaneously compounded intraspinal injections with the bacterial endotoxins test.

    Science.gov (United States)

    Cooper, James F; Thoma, Laura A

    2002-12-15

    Development of a validated bacterial endotoxins test (BET) method for screening extemporaneously compounded intraspinal injections was studied. In accordance with compendial requirements, interference studies were conducted in injectable-grade intraspinal medications using reagents licensed by the Food and Drug Administration. Positive controls were inoculated into a series of decreasing concentrations of each drug (range, 0.05-4 mg/mL) and tested by gel-clot and kinetic-turbidimetric BET methods to determine valid non-interfering concentrations. A simplified BET procedure was used for preparing drug samples, inoculating positive controls, conducting the test, and interpreting the results. The most robust test concentrations were baclofen, bupivacaine, clonidine, and morphine and baclofen, bupivacaine or clonidine. The mixtures were compatible when morphine was diluted to a concentration of < or = 0.25 mg/mL. The substantial dilution required to overcome test inhibition did not compromise the tests sensitivity when compared to the endotoxin limit for intraspinal infusions. Validated BET procedures for intraspinal drugs were sufficiently sensitive to assure the absence of endotoxin contamination.

  13. 1,25-Dihydroxyvitamin D3 promotes a sustained LPS-induced NF-κB-dependent expression of CD55 in human monocytic THP-1 cells.

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    Michael G Izban

    Full Text Available The vitamin D3 system imposes immunosuppressive effects on monocytic cells, in part, by inhibiting NF-κB-dependent expression of proinflammatory mediators. CD55, a cell surface complement regulatory protein that promotes protective and anti-inflammatory properties, is reportedly an NF-κB target gene transiently induced in monocytic cells by the bacterial endotoxin LPS. CD55 is elevated on white cells in women experiencing preterm labor (a pathophysiology commonly associated with bacterial infection and failure to maintain CD55 was associated with subsequent preterm delivery. We examined the influence of vitamin D3 signaling on LPS-induced expression of CD55 in human monocytic THP-1 cells using quantitative PCR, immunoblot, immunohistochemistry, and NF-κB activation pathway inhibitors. Non-NF-κB targets CD14 and CD11b, which modulate bacterial surveillance and eradication, respectively, were also examined. LPS produced a rapid transient 1.6-fold increase in CD55 mRNA. 1,25-D3 alone did not affect CD55 mRNA expression within the first 48 h. However, in 1,25-D3 pretreated cells, LPS produced a >4-fold immediate and sustained increase in CD55 mRNA and protein expression, which was blocked by NF-κB inhibitors. Our results unexpectedly suggest that vitamin D3 signaling may promote an anti-inflammatory response through an NF-κB-dependent increase in CD55 expression. As expected, LPS or 1,25-D3 alone led to sustained increases in CD14 and CD11b expression. In 1,25-D3 pretreated cells, LPS differentially regulated protein expression - CD14 (21-fold increase and CD11b (a transient 2-fold decrease - principally at the posttranscriptional level. The coordinated temporal expression of CD55, CD14 and CD11b would contribute to an anti-inflammatory response by providing protection against complement-mediated cell lysis during pathogen recognition and eradication. Overall, the vitamin D3 system may play a role coordinating an anti-inflammatory response

  14. aged black garlic exerts anti-inflammatory effects by decreasing no and proinflammatory cytokine production with less cytoxicity in LPS-stimulated raw 264.7 macrophages and LPS-induced septicemia mice.

    Science.gov (United States)

    Kim, Min Jee; Yoo, Yung Choon; Kim, Hyun Jung; Shin, Suk Kyung; Sohn, Eun Jeong; Min, A Young; Sung, Nak Yun; Kim, Mee Ree

    2014-10-01

    In this study, the anti-inflammatory and antisepticemic activities of a water extract of aged black garlic (AGE), which is not pungent, were compared with those of raw garlic extract (RGE). The methyl thiazolyl tetrazolium (MTT) assay showed that AGE was not toxic up to 1000 μg/mL and was at least four times less cytotoxic than RGE. AGE significantly suppressed the production of nitric oxide (NO), tumor-necrosis factor-α (TNF-α), and prostaglandin (PG)-E2 in a dose-dependent manner in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Furthermore, the inhibitory effect of AGE on LPS-induced inflammation was confirmed by downregulation of inducible NO synthase and TNF-α mRNA expression, as well as cyclooxygenase-2 protein expression. The anti-inflammatory activities of AGE were similar to those of RGE at nontoxic concentrations up to 250 μg/mL. Signal transduction pathway studies further indicated that both garlic extracts inhibited activation of mitogen-activated protein kinase and nuclear factor-κB induced by LPS stimulation. Treatment with both AGE and RGE in an in vivo experiment of LPS-induced endotoxemia significantly reduced the level of TNF-α and interleukin-6 in serum and completely protected against LPS-induced lethal shock in C57BL/6 mice. The results suggest that AGE is a more promising nutraceutical or medicinal agent to prevent or cure inflammation-related diseases for safety aspects compared with RGE.

  15. Effect of continuous veno-venous hemodiafiltration on endotoxin-induced acute lung injury of the piglets.

    Science.gov (United States)

    Lu, Guo-ping; Gong, Jing-yu; Lu, Zhu-jin; Zhang, Lin-en; Kissoon, Niranjan

    2011-03-01

    To evaluate the effect of continuous veno-venous hemodialysis filtration (CVVHDF) on cardiopulmonary function and clearance of inflammatory mediators in piglets with endotoxin-induced acute lung injury. A randomized controlled trial. An animal laboratory in a tertiary care pediatric center. : Eighteen piglets, weighing 6-8 kg. The piglets were anesthetized, ventilated, and received an intravenous infusion of 150 μg/kg of endotoxin. They were then randomly divided into three groups: control group (n = 6) received Ringer's lactate solution; the heparin group (n = 6) received heparin infusion plus Ringer's lactate solution; and the CVVHDF group (n = 6) received CVVHDF plus heparin infusion and Ringer's lactate solution. Parameters measured simultaneously were: heart rate, mean arterial blood pressure, central venous pressure, pulse contour cardiac index, cardiac function index, left ventricular contractile index, and systemic vascular resistance index; extra vascular lung water index, respiratory rate, dynamic pulmonary compliance, airway resistance, Pao2/Fio2 ratio, serum tumor necrosis factor-α, and soluble tumor necrosis factor receptor. Lung tissue was obtained for pathologic lung injury scoring and wet/dry weight ratio. Endotoxin challenge decreased oxygenation and pulmonary mechanics, suppressed cardiac function, increased extravascular lung water, and elevated serum inflammatory mediators (tumor necrosis factor-α and soluble tumor necrosis factor receptor). After CVVHDF, pulmonary function and oxygenation improved (Pao2/Fio2, 291.5 ± 75.9 vs. 217.2 ± 45.4, respectively, p < .05); arterial blood pressure and cardiac function were restored (pulse contour cardiac index, 3.95 ± 0.52 L/min/m(2) vs. 2.69 ± 0.49 L/min/m(2), respectively, p < .05); extravascular lung water decreased, and serum inflammatory markers also decreased. Lung injury score improved and wet/dry weight ratio decreased. Early CVVHDF has a beneficial effect on acute lung injury in piglets

  16. Effect of Tris-acetate buffer on endotoxin removal from human-like collagen used biomaterials

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Huizhi; Fan, Daidi, E-mail: fandaidi@nwu.edu.cn; Deng, Jianjun; Zhu, Chenghui; Hui, Junfeng; Ma, Xiaoxuan

    2014-09-01

    Protein preparation, which has active ingredients designated for the use of biomaterials and therapeutical protein, is obtained by genetic engineering, but products of genetic engineering are often contaminated by endotoxins. Because endotoxin is a ubiquitous and potent proinflammatory agent, endotoxin removal or depletion from protein is essential for researching any biomaterials. In this study, we have used Tris-acetate (TA) buffer of neutral pH value to evaluate endotoxins absorbed on the Pierce high-capacity endotoxin removal resin. The effects of TA buffer on pH, ionic strength, incubation time as well as human-like collagen (HLC) concentration on eliminating endotoxins are investigated. In the present experiments, we design an optimal method for TA buffer to remove endotoxin from recombinant collagen and use a chromogenic tachypleus amebocyte lysate (TAL) test kit to measure the endotoxin level of HLC. The present results show that, the endotoxins of HLC is dropped to 8.3 EU/ml at 25 mM TA buffer (pH 7.8) with 150 mM NaCl when setting incubation time at 6 h, and HLC recovery is about 96%. Under this experimental condition, it is proved to exhibit high efficiencies of both endotoxin removal and collagen recovery. The structure of treated HLC was explored by Transmission Electron Microscopy (TEM), demonstrating that the property and structure of HLC treated by TA buffer are maintained. Compared to the most widely used endotoxin removal method, Triton X-114 extraction, using TA buffer can obtain the non-toxic HLC without extra treatment for removing the toxic substances in Triton X-114. In addition, the present study aims at establishing a foundation for further work in laboratory animal science and providing a foundation for medical grade biomaterials. - Graphical abstract: The processes of endotoxins adsorbed from HLC. - Highlights: • TA buffer is a mild buffer system for endotoxins removal of HLC. • TA buffer may facilitate endotoxins adsorbed on the

  17. Are cats and dogs the major source of endotoxin in homes?

    Science.gov (United States)

    Ownby, D R; Peterson, E L; Wegienka, G; Woodcroft, K J; Nicholas, C; Zoratti, E; Johnson, C C

    2013-06-01

    Previous studies have suggested that exposure to cats and dogs during early childhood reduces the risk of allergic disease, possibly by increasing home endotoxin exposure. This study asked the question of whether cats and dogs are the dominant influence on dust endotoxin concentrations in homes after considering other variables reportedly associated with endotoxin. The presence of cats or dogs in homes, household and home characteristics, and dust endotoxin concentrations from 5 locations were assessed in 966 urban and suburban homes. Whether considered together as pets or as cats and dogs separately, the presence of cats and dogs significantly contributed to living room and bedroom floor endotoxin concentrations, but not to bed endotoxin concentrations. However, the two variables consistently related to endotoxin in all home sites were the home occupant density (occupants/room) and cleanliness of the home. Our data suggest that reducing occupant density and improving home cleanliness would reduce home endotoxin concentrations more than removing pet cats or dogs from the home. Many studies have shown that early childhood exposure to indoor cats or dogs is associated with a reduced risk of later allergic disease and asthma. An important question is whether alteration in allergic risk associated with cat and dog exposure results from increased endotoxin exposure or from some other associated exposure. Our findings show that cats and dogs are not the dominant source of endotoxin in homes; rather, the density of human occupation and poor cleaning contribute more consistently to higher home endotoxin concentrations especially in the beds. © 2012 John Wiley & Sons A/S. Published by Blackwell Publishing Ltd.

  18. A synthetic cationic antimicrobial peptide inhibits inflammatory response and the NLRP3 inflammasome by neutralizing LPS and ATP.

    Directory of Open Access Journals (Sweden)

    Lan-Hui Li

    Full Text Available Antimicrobial peptides (AMPs are one of the most important defense mechanisms against bacterial infections in insects, plants, non-mammalian vertebrates, and mammals. In the present study, a class of synthetic AMPs was evaluated for anti-inflammatory activity. One cationic AMP, GW-A2, demonstrated the ability to inhibit the expression levels of nitric oxide (NO, inducible NO synthase (iNOS, cyclooxygenase-2 (COX-2, tumor necrosis factor-α (TNF-α and interleukin-6 (IL-6 in lipopolysaccharide (LPS-activated macrophages. GW-A2 reduced LPS-induced increases in the phosphorylation of mitogen-activated protein kinase and protein kinase C-α/δ and the activation of NF-κB. GW-A2 also inhibited NLRP3 inflammasome activation induced by LPS and ATP. Furthermore, in the mice injected with LPS, GW-A2 reduced (1 the concentration of IL-1β, IL-6 and TNF-α in the serum; (2 the concentration of TNF-α in the peritoneal lavage; (3 the expression levels of iNOS, COX-2 and NLRP3 in the liver and lung; (4 the infiltration of polymorphonuclear neutrophils in the liver and lung. The underlying mechanisms for the anti-inflammatory activity of GW-A2 were found to be partially due to LPS and ATP neutralization. These results provide insights into how GW-A2 inhibits inflammation and the NLRP3 inflammasome and provide a foundation for the design of rational therapeutics for inflammation-related diseases.

  19. Serum amyloid P component bound to gram-negative bacteria prevents lipopolysaccharide-mediated classical pathway complement activation

    NARCIS (Netherlands)

    de Haas, CJC; van Leeuwen, EMM; van Bommel, T; Verhoef, J; van Kessel, KPM; van Strijp, JAG

    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

  20. Pantoea agglomerans: a marvelous bacterium of evil and good.Part I. Deleterious effects: Dust-borne endotoxins and allergens - focus on cotton dust.

    Science.gov (United States)

    Dutkiewicz, Jacek; Mackiewicz, Barbara; Lemieszek, Marta Kinga; Golec, Marcin; Milanowski, Janusz

    2015-01-01

    The ubiquitous Gram-negative bacterium Pantoea agglomerans (synonyms: Enterobacter agglomerans, Erwinia herbicola) is known both as an epiphytic microbe developing on the surface of plants and as an endophytic organism living inside the plants. The bacterium occurs also abundantly in plant and animal products, in the body of arthropods and other animals, in water, soil, dust and air, and occasionally in humans. From the human viewpoint, the role of this organism is ambiguous, both deleterious and beneficial: on one side it causes disorders in people exposed to inhalation of organic dusts and diseases of crops, and on the other side it produces substances effective in the treatment of cancer and other diseases of humans and animals, suppresses the development of various plant pathogens, promotes plant growth, and appears as a potentially efficient biofertilizer and bioremediator. P. agglomerans was identified as a predominant bacterium on cotton plant grown all over the world, usually as an epiphyte, rarely as pathogen. It is particularly numerous on cotton bract after senescence. During processing of cotton in mills, bacteria and their products are released with cotton dust into air and are inhaled by workers, causing respiratory and general disorders, usually defined as byssinosis. The most adverse substance is endotoxin, a heteropolymer macromolecule present in the outermost part of the cell wall, consisting of lipopolysaccharide (LPS) as a major constituent, phospholipids and protein. The numerous experiments carried out in last quarter of XXth century on laboratory animals and human volunteers supported a convincing evidence that the inhaled endotoxin produced by P. agglomerans causes numerous pathologic effects similar to those elicited by cotton dust, such as influx of free lung cells into airways and activation of alveolar macrophages which secrete mediators (prostaglandins, platelet-activating factor, interleukin-1, tumor necrosis factor) that cause

  1. [Study on effect and mechanism of cinnabaris and realgar in promoting awake of endotoxin-induced brain injury rat applied with Angong Niuhuang Wan].

    Science.gov (United States)

    Zhu, Kun-Jie; Sun, Jian-Ning

    2014-10-01

    To explore the effect and mechanism of cinnabaris and realgar in promoting awake effect of endotoxin- induced brain injury rat applied with Angong Niuhuang Wan. Normal rats implanted cortical electrode in advance were divided into 6 groups: control, model, the Angong Niuhuang Wan (AGNH, 0.4, 0.2 g · kg(-1)), the Angong Niuhuang Wan without cinnabaris and realgar (QZX-AGNH, 0.32, 0.16 g · kg(-1)). Rats in the control and model groups were given distilled water. After three days of intragastric administration, the brain injury model was injected with endotoxin through tail vein. Then trace electro-corticogram (EcoG) 1-6 h after LPS injection, and compare the power and relative power of beta (β) and delta-waves (δ) at 6 h of these groups. The content of acetylcholine (Ach) and the affinity of M-receptor (M-R) in cortex and brainstem were detected by alkaline hydroxylamine colorimetric method and radioactive ligand binding assay, respectively. AGNH (0.4, 0.2 g · kg(-1)) could increase the power and relative power of β and AGNH (0.4 g · kg(-1)) showed better action on brain electrical activation. QZX-AGNH showed weak effect on it. AGNH (0.4 g · kg(-1)) could increase the affinity of M-R in cortex and the content of Ach in brainstem. The action of QZX-AGNH was not obvious. In endotoxin-induced brain injury rats, AGNH can raise the cholinergic system function of cortex, and strengthen the uplink of cortex activation of brainstem cholinergic system, improve the level of cortical activity and enhance the activation of EcoG to promote the body's awakening. QZX-AGNH show weak effect. Cinnabaris and realgar play an important role in promoting awake effect in endotoxin-induced brain injury applied with Angong Niuhuang Wan. The mechanism may be related to cortical and brainstem cholinergic system function.

  2. Pantoea agglomerans: a marvelous bacterium of evil and good.Part I. Deleterious effects: Dust-borne endotoxins and allergens – focus on cotton dust

    Directory of Open Access Journals (Sweden)

    Jacek Dutkiewicz

    2015-12-01

    Full Text Available The ubiquitous Gram-negative bacterium Pantoea agglomerans (synonyms: Enterobacter agglomerans, Erwinia herbicola is known both as an epiphytic microbe developing on the surface of plants and as an endophytic organism living inside the plants. The bacterium occurs also abundantly in plant and animal products, in the body of arthropods and other animals, in water, soil, dust and air, and occasionally in humans. From the human viewpoint, the role of this organism is ambiguous, both deleterious and beneficial: on one side it causes disorders in people exposed to inhalation of organic dusts and diseases of crops, and on the other side it produces substances effective in the treatment of cancer and other diseases of humans and animals, suppresses the development of various plant pathogens, promotes plant growth, and appears as a potentially efficient biofertilizer and bioremediator. P. agglomerans was identified as a predominant bacterium on cotton plant grown all over the world, usually as an epiphyte, rarely as pathogen. It is particularly numerous on cotton bract after senescence. During processing of cotton in mills, bacteria and their products are released with cotton dust into air and are inhaled by workers, causing respiratory and general disorders, usually defined as byssinosis. The most adverse substance is endotoxin, a heteropolymer macromolecule present in the outermost part of the cell wall, consisting of lipopolysaccharide (LPS as a major constituent, phospholipids and protein. The numerous experiments carried out in last quarter of XXth century on laboratory animals and human volunteers supported a convincing evidence that the inhaled endotoxin produced by P. agglomerans causes numerous pathologic effects similar to those elicited by cotton dust, such as influx of free lung cells into airways and activation of alveolar macrophages which secrete mediators (prostaglandins, platelet-activating factor, interleukin-1, tumor necrosis factor

  3. 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, therec....html) (.csml) Show Function of lipopolysaccharide (LPS)-binding protein (LBP) and CD14, thereceptor for LPS.../LBP complexes: a short review. PubmedID 1373512 Title Function of lipopolysaccharide

  4. AMPK activation ameliorates D-GalN/LPS-induced acute liver failure by upregulating Foxo3A to induce autophagy.

    Science.gov (United States)

    Liu, Yan-Min; Lv, Jun; Zeng, Qing-Lei; Shen, Shen; Xing, Ji-Yuan; Zhang, Ying-Ying; Zhang, Zhi-Hao; Yu, Zu-Jiang

    2017-09-15

    Acute liver failure (ALF) is an uncommon but serious disease still carrying a high mortality. This study aimed to investigate the mechanism of AMPK on D-GalN/LPS-induced ALF. In this study, we utilized intraperitoneal injection of D-GalN/LPS to induce ALF model, and analyzed the expression of AMPK, inflammatory cytokines (TNF-α, IL-1β and IL-6), Foxo3A and autophagy-related genes (Atg-5, Beclin-1, Atg-7) by real-time quantitative polymerase chain reaction (RT-PCR) in liver tissue. We also examined the level of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum of ALF mice. AMPK activation and inhibition of autophagy were induced by AICAR and 3-MA, respectively. Silence and overexpression of Foxo3A were performed by si-Foxo3A and pcDNA-Foxo3A, respectively. Lastly, the BMDM-conditioned medium (BMDM-CM) derived from BMDMs treated with AICAR and LPS were used to explore the effect of AMPK and Foxo3A on hepatocytes. The expression of AMPK was decreased in liver tissue and the level of ALT and AST were increased in serum of D-GalN/LPS-induced ALF mice. AMPK activation ameliorated ALF by inhibiting inflammation (downregulated TNF-α, IL-1β and IL-6 expression), activating autophagy (increased Atg-5, Beclin-1 and Atg-7 expression) and upregulating Foxo3A expression. Silence of Foxo3A decreased AMPK-activated autophagy, but overexpressing Foxo3A attenuated liver failure by activating autophagy. In addition, AMPK activation alleviated liver failure in vitro. Thus, AMPK/Foxo3A/autophagy pathway may be an effective treatment approach to ameliorate ALF. Copyright © 2017 Elsevier Inc. All rights reserved.

  5. Cardiorespiratory control and cytokine profile in response to heat stress, hypoxia, and lipopolysaccharide (LPS) exposure during early neonatal period.

    Science.gov (United States)

    McDonald, Fiona B; Chandrasekharan, Kumaran; Wilson, Richard J A; Hasan, Shabih U

    2016-02-01

    Sudden infant death syndrome (SIDS) is one of the most common causes of postneonatal infant mortality in the developed world. An insufficient cardiorespiratory response to multiple environmental stressors (such as prone sleeping positioning, overwrapping, and infection), during a critical period of development in a vulnerable infant, may result in SIDS. However, the effect of multiple risk factors on cardiorespiratory responses has rarely been tested experimentally. Therefore, this study aimed to quantify the independent and possible interactive effects of infection, hyperthermia, and hypoxia on cardiorespiratory control in rats during the neonatal period. We hypothesized that lipopolysaccharide (LPS) administration will negatively impact cardiorespiratory responses to increased ambient temperature and hypoxia in neonatal rats. Sprague-Dawley neonatal rat pups were studied at postnatal day 6-8. Rats were examined at an ambient temperature of 33°C or 38°C. Within each group, rats were allocated to control, saline, or LPS (200 μg/kg) treatments. Cardiorespiratory and thermal responses were recorded and analyzed before, during, and after a hypoxic exposure (10% O2). Serum samples were taken at the end of each experiment to measure cytokine concentrations. LPS significantly increased cytokine concentrations (such as TNFα, IL-1β, MCP-1, and IL-10) compared to control. Our results do not support a three-way interaction between experimental factors on cardiorespiratory control. However, independently, heat stress decreased minute ventilation during normoxia and increased the hypoxic ventilatory response. Furthermore, LPS decreased hypoxia-induced tachycardia. Herein, we provide an extensive serum cytokine profile under various experimental conditions and new evidence that neonatal cardiorespiratory responses are adversely affected by dual interactions of environmental stress factors. © 2016 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on

  6. Mice lacking Tbk1 activity exhibit immune cell infiltrates in multiple tissues and increased susceptibility to LPS-induced lethality.

    Science.gov (United States)

    Marchlik, Erica; Thakker, Paresh; Carlson, Thaddeus; Jiang, Zhaozhao; Ryan, Mark; Marusic, Suzana; Goutagny, Nadege; Kuang, Wen; Askew, G Roger; Roberts, Victoria; Benoit, Stephen; Zhou, Tianhui; Ling, Vincent; Pfeifer, Richard; Stedman, Nancy; Fitzgerald, Katherine A; Lin, Lih-Ling; Hall, J Perry

    2010-12-01

    TBK1 is critical for immunity against microbial pathogens that activate TLR4- and TLR3-dependent signaling pathways. To address the role of TBK1 in inflammation, mice were generated that harbor two copies of a mutant Tbk1 allele. This Tbk1(Δ) allele encodes a truncated Tbk1(Δ) protein that is catalytically inactive and expressed at very low levels. Upon LPS stimulation, macrophages from Tbk1(Δ/Δ) mice produce normal levels of proinflammatory cytokines (e.g., TNF-α), but IFN-β and RANTES expression and IRF3 DNA-binding activity are ablated. Three-month-old Tbk1(Δ/Δ) mice exhibit mononuclear and granulomatous cell infiltrates in multiple organs and inflammatory cell infiltrates in their skin, and they harbor a 2-fold greater amount of circulating monocytes than their Tbk1(+/+) and Tbk1(+/Δ) littermates. Skin from 2-week-old Tbk1(Δ/Δ) mice is characterized by reactive changes, including hyperkeratosis, hyperplasia, necrosis, inflammatory cell infiltrates, and edema. In response to LPS challenge, 3-month-old Tbk1(Δ/Δ) mice die more quickly and in greater numbers than their Tbk1(+/+) and Tbk1(+/Δ) counterparts. This lethality is accompanied by an overproduction of several proinflammatory cytokines in the serum of Tbk1(Δ/Δ) mice, including TNF-α, GM-CSF, IL-6, and KC. This overproduction of serum cytokines in Tbk1(Δ/Δ) mice following LPS challenge and their increased susceptibility to LPS-induced lethality may result from the reactions of their larger circulating monocyte compartment and their greater numbers of extravasated immune cells.

  7. Enhancement of systemic and sputum granulocyte response to inhaled endotoxin in people with the GSTM1 null genotype

    Science.gov (United States)

    To determine if the GSTM1 null genotype is a risk factor for increased inflammatory response to inhaled endotoxin. Methods 35 volunteers who had undergone inhalation challenge with a 20 000 endotoxin unit dose of Clinical Center Reference Endotoxin (CCRE) were genotyped for the G...

  8. DMPD: Gram-negative endotoxin: an extraordinary lipid with profound effects oneukaryotic signal transduction. [Dynamic Macrophage Pathway CSML Database

    Lifescience Database Archive (English)

    Full Text Available 1916089 Gram-negative endotoxin: an extraordinary lipid with profound effects oneuk...ep;5(12):2652-60. (.png) (.svg) (.html) (.csml) Show Gram-negative endotoxin: an extraordinary lipid with profound effect...tive endotoxin: an extraordinary lipid with profound effects oneukaryotic signal transduction. Authors Raetz

  9. Ambient Endotoxin Concentrations and Assessment of Offsite Transport at Open-Lot and Open-Freestall Dairies.

    Science.gov (United States)

    Endotoxins are derived from gram-negative bacteria and are a potent inducer of inflammatory reactions in the respiratory tract when inhaled. To assess daily fluctuations of airborne endotoxin and their potential for transport from dairies, endotoxin concentrations were monitored over an 8-h period a...

  10. Comparative Study of Plasma Endotoxin with Procalcitonin Levels in Diagnosis of Bacteremia in Intensive Care Unit Patients

    Directory of Open Access Journals (Sweden)

    Tao Wang

    2016-01-01

    Conclusions: Plasma endotoxin was more related to GN than to Gram-positive bacteremia, and that endotoxin level was species dependent, but PCT level remained relatively more stable within the GN bacteria caused bacteremia. Both GN and positive bacteria caused bacteremia in the ICU patients in different regions of China. And PCT is a more valuable biomarker than endotoxin in the diagnosis of bacteremia.

  11. Effect of Escherichia coli endotoxin on Archachatina marginata hemolymph coagulation system.

    Science.gov (United States)

    Salawu, Musa O; Oloyede, Oyelola B; Oladiji, Adenike T; Muhammad, Nasiru O; Yakubu, Musa T

    2011-10-01

    Archachatina marginata Swainson (Achatinidae) is found in Nigeria, West Africa. Its hemolymph is applied as a disinfectant to blades and fresh cuts of circumcision in Yorubaland. The hemolymph is also used in traditional medicine practice. Investigation into its anti-endotoxin response is being studied for the first time. This study determined whether endotoxin causes measurable and concentration-dependent protein coagulation in the separate hemolymph fractions and in hemocyte lysate (HL)/plasma mixtures. Endotoxin was prepared by inoculating 5% w/v dextrose with locally isolated Escherichia coli cells and incubated for 48 h before sterilization. Pyrogenicity was determined by rabbit test method and use the of LAL kit. Hemolymph fractions were exposed to endotoxin while controls were exposed to endotoxin-free water (0.025 EU/ml). HL/plasma (1:1 v/v) was exposed to varied endotoxin concentrations. Data indicated significantly higher protein coagulates induced by endotoxin in all the hemolymph fractions (P marginata. This finding may be employed in detection and quantification of endotoxin in future.

  12. Experimental design and Bayesian networks for enhancement of delta-endotoxin production by Bacillus thuringiensis.

    Science.gov (United States)

    Ennouri, Karim; Ayed, Rayda Ben; Hassen, Hanen Ben; Mazzarello, Maura; Ottaviani, Ennio

    2015-12-01

    Bacillus thuringiensis (Bt) is a Gram-positive bacterium. The entomopathogenic activity of Bt is related to the existence of the crystal consisting of protoxins, also called delta-endotoxins. In order to optimize and explain the production of delta-endotoxins of Bacillus thuringiensis kurstaki, we studied seven medium components: soybean meal, starch, KH₂PO₄, K₂HPO₄, FeSO₄, MnSO₄, and MgSO₄and their relationships with the concentration of delta-endotoxins using an experimental design (Plackett-Burman design) and Bayesian networks modelling. The effects of the ingredients of the culture medium on delta-endotoxins production were estimated. The developed model showed that different medium components are important for the Bacillus thuringiensis fermentation. The most important factors influenced the production of delta-endotoxins are FeSO₄, K2HPO₄, starch and soybean meal. Indeed, it was found that soybean meal, K₂HPO₄, KH₂PO₄and starch also showed positive effect on the delta-endotoxins production. However, FeSO4 and MnSO4 expressed opposite effect. The developed model, based on Bayesian techniques, can automatically learn emerging models in data to serve in the prediction of delta-endotoxins concentrations. The constructed model in the present study implies that experimental design (Plackett-Burman design) joined with Bayesian networks method could be used for identification of effect variables on delta-endotoxins variation.

  13. Personal exposure to dust and endotoxin in Robusta and Arabica coffee processing factories in Tanzania.

    Science.gov (United States)

    Sakwari, Gloria; Mamuya, Simon H D; Bråtveit, Magne; Larsson, Lennart; Pehrson, Christina; Moen, Bente E

    2013-03-01

    Endotoxin exposure associated with organic dust exposure has been studied in several industries. Coffee cherries that are dried directly after harvest may differ in dust and endotoxin emissions to those that are peeled and washed before drying. The aim of this study was to measure personal total dust and endotoxin levels and to evaluate their determinants of exposure in coffee processing factories. Using Sidekick Casella pumps at a flow rate of 2l/min, total dust levels were measured in the workers' breathing zone throughout the shift. Endotoxin was analyzed using the kinetic chromogenic Limulus amebocyte lysate assay. Separate linear mixed-effects models were used to evaluate exposure determinants for dust and endotoxin. Total dust and endotoxin exposure were significantly higher in Robusta than in Arabica coffee factories (geometric mean 3.41 mg/m(3) and 10 800 EU/m(3) versus 2.10 mg/m(3) and 1400 EU/m(3), respectively). Dry pre-processed coffee and differences in work tasks explained 30% of the total variance for total dust and 71% of the variance for endotoxin exposure. High exposure in Robusta processing is associated with the dry pre-processing method used after harvest. Dust and endotoxin exposure is high, in particular when processing dry pre-processed coffee. Minimization of dust emissions and use of efficient dust exhaust systems are important to prevent the development of respiratory system impairment in workers.

  14. Effect of endotoxin tolerance on drug hepatotoxicity: amelioration of taurolithocholate cholestasis in the perfused rat liver.

    Science.gov (United States)

    Utili, R; Adinolfi, L E; Gaeta, G B; Tripodi, M F; Alvaro, D

    1987-01-01

    Induction of endotoxin tolerance may cause resistance not only to endotoxin itself but also to the hepatotoxic effects of other membrane-active agents. To further study this effect, we tested whether endotoxin tolerance could ameliorate the adverse effects of taurolithocholate (TLCA) which causes cholestasis by altering liver plasma membrane organization. Isolated perfused rat livers from endotoxin-tolerant rats had a lower basal bile flow than control livers. However, a bolus addition of TLCA at 3 X 10(-5) or 5 X 10(-5) M in the perfusate caused a marked and prolonged decrease of bile flow in controls, but only a transient and significantly less pronounced diminution of bile flow in endotoxin-tolerant livers. Likewise, TLCA caused a significantly lower alteration of hepatocyte membrane permeability, as measured by sucrose permeability studies, in endotoxin-tolerant livers than in controls. Analysis of bile acid composition of bile from endotoxin-tolerant livers demonstrated that they excreted greater amounts of total bile acids, in particular TLCA and taurocholate, than controls. These results demonstrated a protective effect of endotoxin-tolerance against TLCA toxicity which may result from an altered interaction of TLCA with liver membranes and an increased clearance of TLCA.

  15. An Endotoxin Tolerance Signature Predicts Sepsis and Organ Dysfunction at Initial Clinical Presentation

    Directory of Open Access Journals (Sweden)

    Olga M. Pena

    2014-11-01

    Interpretation: Our data support an updated model of sepsis pathogenesis in which endotoxin tolerance-mediated immune dysfunction (cellular reprogramming is present throughout the clinical course of disease and related to disease severity. Thus endotoxin tolerance might offer new insights guiding the development of new therapies and diagnostics for early sepsis.

  16. A comparative study of different strategies for removal of endotoxins from bacteriophage preparations.

    Science.gov (United States)

    Van Belleghem, Jonas D; Merabishvili, Maya; Vergauwen, Bjorn; Lavigne, Rob; Vaneechoutte, Mario

    2017-01-01

    Bacterial endotoxins have high immunogenicity. Phage biology studies as well as therapeutic phage applications necessitate highly purified phage particles. In this study, we compared combinations of seven different endotoxin removal strategies and validated their endotoxin removal efficacy for five different phages (i.e. four Pseudomonas aeruginosa phages and one Staphylococcus aureus phage). These purification strategies included Endotrap HD column purification and/or CsCl density centrifugation in combination with Endotrap purification, followed by organic solvent (1-octanol), detergent (Triton X-100), enzymatic inactivation of the endotoxin using alkaline phosphatase and CIM monolytic anion exchange chromatography. We show that CsCl density purification of the P. aeruginosa phages, at an initial concentration of 1012-1013pfu/ml, led to the strongest reduction of endotoxins, with an endotoxin removal efficacy of up to 99%, whereas additional purification methods did not result in a complete removal of endotoxins from the phage preparations and only yielded an additional endotoxin removal efficacy of 23 to 99%, sometimes accompanied with strong losses in phage titer. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Development of an in vitro Endotoxin Test for Monoolein–Water ...

    African Journals Online (AJOL)

    Erah

    vitro endotoxin test (LAL test) for water- insoluble gel products. Since our research group is currently developing monoolein–water liquid crystalline gels of gentamycin for use as bioresorbable implants for the treatment of chronic osteomyelitis [10-12], we set out to develop an in vitro test method for endotoxin in this.

  18. Single session of Nd:YAG laser intracanal irradiation neutralizes endotoxin in dental root dentin

    Science.gov (United States)

    Archilla, José R. F.; Moreira, Maria S. N. A.; Miyagi, Sueli P. H.; Bombana, Antônio C.; Gutknecht, Norbert; Marques, Márcia M.

    2012-11-01

    Endotoxins released in the dental root by Gram-negative microorganisms can be neutralized by calcium hydroxide, when this medication is applied inside the root canal for at least seven days. However, several clinical situations demand faster root canal decontamination. Thus, for faster endotoxin neutralization, endodontists are seeking additional treatments. The in vitro study tested whether or not intracanal Nd:YAG laser irradiation would be able to neutralize endotoxin within the human dental root canal in a single session. Twenty-four human teeth with one root were mounted between two chambers. After conventional endodontic treatment, root canals were contaminated with Escherichia coli endotoxin. Then they were irradiated or not (controls) in contact mode with an Nd:YAG laser (1.5 W, 15 Hz, 100 mJ and pulse fluency of 124 J/cm2). The endotoxin activity was measured using the limulus lysate technique and data were statistically compared (p≤0.05). The concentration of active endotoxin measured in the negative control group was significantly lower than that of the positive control group (p=0.04). The concentrations of endotoxin in both irradiated groups were significantly lower than that of the positive control group (p=0.027) and similar to that of negative control group (p=0.20). A single session of intracanal Nd:YAG laser irradiation is able to neutralize endotoxin in the dental root tissues.

  19. ENDOTOXIN RELEASE AND TUMOR-NECROSIS-FACTOR FORMATION DURING CARDIOPULMONARY BYPASS

    NARCIS (Netherlands)

    JANSEN, NJG; VANOEVEREN, W; GU, YJ; VANVLIET, MH; EIJSMAN, L; WILDEVUUR, CRH

    1992-01-01

    Endotoxin, when released into the systemic circulation during cardiopulmonary bypass (CPB), might induce activation of plasmatic systems and blood cells during CPB, in addition to a material-dependent blood activation during CPB. However, the role of endotoxin in the development of this so-called

  20. Endotoxin testing of proteins for parenteral administration using the Mono Mac 6 assay

    DEFF Research Database (Denmark)

    Moesby, Lise; Hansen, E W; Christensen, J D

    2000-01-01

    Pharmaceutical products containing proteins cause problems in testing for endotoxin and pyrogens. Many proteins interfere with the LAL test and the proteins are immunogenic in rabbits. The monocytic cell line Mono Mac 6 is an alternative assay for detection of endotoxin and other pyrogens....

  1. Systemic anti-tumor necrosis factor antibody treatment exacerbates endotoxin-induced uveitis in the rat

    NARCIS (Netherlands)

    de Vos, A. F.; van Haren, M. A.; Verhagen, C.; Hoekzema, R.; Kijlstra, A.

    1995-01-01

    Tumor necrosis factor is released in the circulation and aqueous humor during endotoxin-induced uveitis, and induces acute uveitis when injected intraocularly in rats. To elucidate the role of tumor necrosis factor in the development of endotoxin-induced uveitis we analysed the effect of

  2. Effect of extraction and assay media on analysis of airborne endotoxin

    NARCIS (Netherlands)

    Spaan, S.; Doekes, G.; Heederik, D.; Thorne, P.S.; Wouters, I.M.

    2008-01-01

    The measurement of airborne endotoxins is thus far not standardized. Earlier studies reported higher endotoxin yields when Tween 20 was added to the media used for filter extraction and in the Limulus amebocyte lysate (LAL) assay. This study compared four common media and assessed the effects of

  3. Prenatal exposure to a pro-inflammatory stimulus causes delays in the development of the innate immune response to LPS in the offspring.

    Science.gov (United States)

    Hodyl, Nicolette A; Krivanek, Klara M; Lawrence, Elizabeth; Clifton, Vicki L; Hodgson, Deborah M

    2007-10-01

    Growing evidence suggests that maternal health during the prenatal period is a critical determinant of adult immuno-competence. This study aimed to characterise the innate immune response to bacterial exposure in rat offspring following maternal exposure to a pro-inflammatory stimulus. The offspring's innate immune responses were investigated at four developmental timepoints in the rat by determination of immune cell subtypes and TNF-alpha and IL-1beta response to in-vivo LPS exposure. The pre-weaned offspring of exposed dams demonstrated no immune response to the LPS challenge, whereas control offspring responded with a typical elevation in cytokine levels. In pubescence no differences were observed between the responses of the control and exposed offspring. In adulthood and senescence, offspring of endotoxin treated dams had significantly less monocytes in circulation than control offspring and differential sex effects were only evident in these older animals. The developmental profile of immune functioning following prenatal immune activation has not previously been demonstrated. This study highlights the prenatal period as one of importance in determining later immune function.

  4. Endotoxemia por lipopolissacarídeo de Escherichia coli, em eqüinos: efeitos de antiinflamatórios nas concentrações sérica e peritoneal do fator de necrose tumoral alfa (TNF-alfa Escherichia coli lipopolisacharide (LPS induced endotoxemia in horses: effects of anti-inflammatory drugs on seric and peritoneal tumor necrosis factor alpha (TNF-alpha concentrations

    Directory of Open Access Journals (Sweden)

    R.C. Campebell

    2007-08-01

    . Control group horses received 9% sodium chloride, IV. SD group animals were orally administrated 2.2mg/kg sodium diclofenac and DM horses received 1.1mg/kg dexamethazone, IV, 30 and 60 minutes before endotoxin infusion, respectively. TNF-alpha concentration was measured in serum and peritoneal fluid by toxicity assay using L929 lineage cells at 0, 1¼, 3 and 6 hours after LPS injection. Ninety minutes after endotoxemy induction, it was verified a significant increase of serum TNF-a concentration in horses from control group in relation to the basal values as well as results of horses from SD and DM groups. In peritoneal fluid, the measured concentrations were lower than those from TNF-a standard curve and difference among the groups was not verified (P>0.05.

  5. Inhibition of Neuroinflammation in LPS-Activated Microglia by Cryptolepine

    Directory of Open Access Journals (Sweden)

    Olumayokun A. Olajide

    2013-01-01

    Full Text Available Cryptolepine, an indoloquinoline alkaloid in Cryptolepis sanguinolenta, has anti-inflammatory property. In this study, we aimed to evaluate the effects of cryptolepine on lipopolysaccharide (LPS- induced neuroinflammation in rat microglia and its potential mechanisms. Microglial activation was induced by stimulation with LPS, and the effects of cryptolepine pretreatment on microglial activation and production of proinflammatory mediators, PGE2/COX-2, microsomal prostaglandin E2 synthase and nitric oxide/iNOS were investigated. We further elucidated the role of Nuclear Factor-kappa B (NF-κB and the mitogen-activated protein kinases in the antiinflammatory actions of cryptolepine in LPS-stimulated microglia. Our results showed that cryptolepine significantly inhibited LPS-induced production of tumour necrosis factor-alpha (TNFα, interleukin-6 (IL-6, interleukin-1beta (IL-1β, nitric oxide, and PGE2. Protein and mRNA levels of COX-2 and iNOS were also attenuated by cryptolepine. Further experiments on intracellular signalling mechanisms show that IκB-independent inhibition of NF-κB nuclear translocation contributes to the anti-neuroinflammatory actions of cryptolepine. Results also show that cryptolepine inhibited LPS-induced p38 and MAPKAPK2 phosphorylation in the microglia. Cell viability experiments revealed that cryptolepine (2.5 and 5 μM did not produce cytotoxicity in microglia. Taken together, our results suggest that cryptolepine inhibits LPS-induced microglial inflammation by partial targeting of NF-κB signalling and attenuation of p38/MAPKAPK2.

  6. A new method for concentration analysis of bacterial endotoxins in perfluorocarbon

    Science.gov (United States)

    Chen, Dan-Dan; Feng, Xiao-Ming; Wang, Chun-Ren; Huang, Qing-Quan; Yang, Zhao-Peng; Meng, Qing-Yuan

    2014-12-01

    This communication demonstrates the feasibility of the gel-clot method for the analysis of bacterial endotoxins in water extracts of perfluorocarbon which is a water insoluble liquid medical device. Perfluorocarbon (10 mL) was shaken with 10mL water for 15 min at 2000 r/min and the endotoxin present was extracted to the aqueous phase without interference inhibition/enhancement of the product and the recovery of endotoxin added to perfluorocarbon was determined. A validation study confirmed that endotoxins presented in perfluorocarbon pass over into the aqueous phase at concentrations of 20, 10 and 5 EU/mL with recoveries from 86.8% to 96.8%. Therefore, the gel-clot test is suitable for detecting bacterial endotoxins in perfluorocarbon which is a water insoluble medical device.

  7. Short time bacterial endotoxins test for positron emission tomography by means of positively charged filters

    Energy Technology Data Exchange (ETDEWEB)

    Nakazawa, Nobuhiro; Wakita, Kazuo [Nishijin Hospital, Kyoto (Japan); Mineura, Katsuyoshi [Kyoto Prefectural Univ. of Medicine (Japan)] (and others)

    2002-11-01

    Positron emission tomography (PET) radiotracers have very short physical half-lives. It is hard to complete a bacterial endotoxins test prior to release from medical institutes. For endotoxin quantitative determination, limulus amebocyte lysate (LAL) reagent and kinetic-turbidimetry system were previously developed. We investigated the possibility of a short time test by means of positively charged filters. As a result of this study, the effects of positively charged filters on endotoxin removal were over 99.5% for [{sup 18}F]FDG and [{sup 18}F]NaF, which were contaminated with the indicated concentration of endotoxin. Combining this filter and the kinetic-turbidimetric method, it was possible to complete a bacterial endotoxins test in 5 min prior to the patient's administration. This test should be required prior to release for PET radiopharmaceutical quality control. It has been suggested that this combination is a good method for this purpose. (author)

  8. Insoluble glycogen, a metabolizable internal adsorbent, decreases the lethality of endotoxin shock in rats

    Directory of Open Access Journals (Sweden)

    S. Sipka

    1997-01-01

    Full Text Available Insoluble glycogen is an enzymatically modified form of naturally occurring soluble glycogen with a great adsorbing capacity. It can be metabolized by phagocytes to glucose. In this study we used insoluble glycogen intravenously in the experimental endotoxin shock of rats. Wistar male rats were sensitized to endotoxin by Pb acetate. The survival of rats were compared in groups of animals endotoxin shock treated and non-treated with insoluble glycogen. Furthermore, we have determined in vitro the binding capacity of insoluble glycogen for endotoxin, tumour necrosis factor alpha, interleukin-1 and secretable phospholipase A2. Use of 10 mg/kg dose of insoluble glycogen could completely prevent the lethality of shock induced by LD50 quantity of endotoxin in rats. All animals treated survived. Insoluble glycogen is a form of ‘metabolizable internal adsorbents’. It can potentially be used for treatment of septic shock.

  9. The Protective Effect of Apamin on LPS/Fat-Induced Atherosclerotic Mice

    Directory of Open Access Journals (Sweden)

    Soo-Jung Kim

    2012-01-01

    Full Text Available Apamin, a peptide component of bee venom (BV, has anti-inflammatory properties. However, the molecular mechanisms by which apamin prevents atherosclerosis are not fully understood. We examined the effect of apamin on atherosclerotic mice. Atherosclerotic mice received intraperitoneal (ip injections of lipopolysaccharide (LPS, 2 mg/kg to induce atherosclerotic change and were fed an atherogenic diet for 12 weeks. Apamin (0.05 mg/kg was administered by ip injection. LPS-induced THP-1-derived macrophage inflammation treated with apamin reduced expression of tumor necrosis factor (TNF-α, vascular cell adhesion molecule (VCAM-1, and intracellular cell adhesion molecule (ICAM-1, as well as the nuclear factor kappa B (NF-κB signaling pathway. Apamin decreased the formation of atherosclerotic lesions as assessed by hematoxylin and elastic staining. Treatment with apamin reduced lipids, Ca2+ levels, and TNF-α in the serum from atherosclerotic mice. Further, apamin significantly attenuated expression of VCAM-1, ICAM-1, TGF-β1, and fibronectin in the descending aorta from atherosclerotic mice. These results indicate that apamin plays an important role in monocyte/macrophage inflammatory processing and may be of potential value for preventing atherosclerosis.

  10. In vivo relevance of polymorphic Interleukin 8 promoter haplotype for the systemic immune response to LPS in Holstein-Friesian calves.

    Science.gov (United States)

    Stojkovic, Bojan; McLoughlin, Rachel M; Meade, Kieran G

    2016-12-01

    Interleukin 8, also known as CXC chemokine ligand 8 (CXCL8), is a critical chemokine in the recruitment of leukocytes to sites of infection and is a potent mediator of inflammation. We previously discovered 29 polymorphic sites in the promoter region of the bovine Interleukin 8 gene, which segregate into two distinct haplotypes, denoted IL8-h1 and IL8-h2. Population genetic analysis of these two haplotypes showed significant inter-breed differences in haplotype frequency, which is suggestive of selection acting at this locus. Furthermore functional characterisation identified that IL8-h2 was more active in mammary epithelial cells stimulated with the bacterial endotoxin, LPS. However, the in vivo relevance of these functional differences in the IL8 gene has not been ascertained. Therefore, in the current study, we tested the hypothesis that IL8 haplotype would result in variation in the systemic immune response to LPS challenge in Holstein-Friesian (HF) calves. A Taqman assay was designed to genotype both Jersey and HF calves, from which 20 healthy HF calves (representing IL8-h1 and IL8-h2) from the same farm were subjected to LPS stimulation via jugular venepuncture (100ng/kg). Systemic immune profiling was subsequently performed up to 216h post-challenge. Haematological analysis showed perturbations in leukocyte populations of cells but only the lymphocyte response was significantly different between IL8-h1 and IL8-h2. IL8 expression levels were significantly different between haplogroups, at both the gene expression and protein levels (P<0.05). Circulating neutrophils were subsequently purified from each haplogroup to measure potential haplotype specific effects on neutrophil migration and bacterial killing but no significant differences were detected, which is likely due to the low circulating levels of IL8. We conclude that IL8 haplotype significantly affects IL8 expression profile in response to bacterial endotoxin in vivo, and the significant increase in IL8

  11. LPS-Stimulated Whole Blood Cytokine Production Is Not Related to Disease Behavior in Patients with Quiescent Crohn's Disease.

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    Mark M T J Broekman

    Full Text Available Crohn's disease (CD is a chronic inflammatory disease in which cytokines play a pivotal role in the induction and maintenance of inflammation. Innate cytokine production is genetically determined and varies largely between individuals; this might impact the severity of inflammation. We aimed to assess whether ex-vivo endotoxin-stimulated levels of cytokines could be associated with disease phenotype.Patients with quiescent CD (Harvey-Bradshaw Index ≤ 4 and negative inflammation markers who were not using immunomodulating drugs or biologicals were eligible. Historical disease characteristics (localization, behavior, number of bowel resections, drug history, extra-intestinal symptoms were extracted from medical records. We measured cytokine levels (tumor necrosis factor (TNF-α, interleukin (IL-1β, IL-6 and IL-10 in supernatants of lipopolysaccharide (LPS -stimulated whole blood cultures and correlated these with disease characteristics and age- and sex-matched healthy controls. In addition, we analyzed whether single nucleotide polymorphisms (SNPs in the promoter region of the TNF-α gene were related to TNF-α levels.We included 75 patients with CD and 24 healthy controls. Six patients were excluded because of increased inflammation markers resulting in a total of 69 patients. The mean age (SD of patients with CD was 51.2 (12.3 years with a mean (SD disease duration of 24.1 (11.5 years. Disease localization, peri-anal involvement and behavior were not related to LPS-stimulated TNF-α, IL-1β, IL-6 or IL-10 levels. In addition, combination of localization with behavior to differentiate mild from severe disease type showed no significant differences. TNF-α levels were higher in patients with CD (428 pg/ml IQR [267-468] compared to healthy controls (459 pg/ml IQR [364-570], p=0.02. We found no associations between SNPs in the promoter region and TNF-α levels.In this study, innate cytokine production of TNF-α, IL-1β, IL-6 and IL-10 was not

  12. LPS-Stimulated Whole Blood Cytokine Production Is Not Related to Disease Behavior in Patients with Quiescent Crohn's Disease.

    Science.gov (United States)

    Broekman, Mark M T J; Roelofs, Hennie M J; Hoentjen, Frank; Wiegertjes, Renske; Stoel, Nicole; Joosten, Leo A; de Jong, Dirk J; Wanten, Geert J A

    2015-01-01

    Crohn's disease (CD) is a chronic inflammatory disease in which cytokines play a pivotal role in the induction and maintenance of inflammation. Innate cytokine production is genetically determined and varies largely between individuals; this might impact the severity of inflammation. We aimed to assess whether ex-vivo endotoxin-stimulated levels of cytokines could be associated with disease phenotype. Patients with quiescent CD (Harvey-Bradshaw Index ≤ 4 and negative inflammation markers) who were not using immunomodulating drugs or biologicals were eligible. Historical disease characteristics (localization, behavior, number of bowel resections, drug history, extra-intestinal symptoms) were extracted from medical records. We measured cytokine levels (tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6 and IL-10) in supernatants of lipopolysaccharide (LPS) -stimulated whole blood cultures and correlated these with disease characteristics and age- and sex-matched healthy controls. In addition, we analyzed whether single nucleotide polymorphisms (SNPs) in the promoter region of the TNF-α gene were related to TNF-α levels. We included 75 patients with CD and 24 healthy controls. Six patients were excluded because of increased inflammation markers resulting in a total of 69 patients. The mean age (SD) of patients with CD was 51.2 (12.3) years with a mean (SD) disease duration of 24.1 (11.5) years. Disease localization, peri-anal involvement and behavior were not related to LPS-stimulated TNF-α, IL-1β, IL-6 or IL-10 levels. In addition, combination of localization with behavior to differentiate mild from severe disease type showed no significant differences. TNF-α levels were higher in patients with CD (428 pg/ml IQR [267-468]) compared to healthy controls (459 pg/ml IQR [364-570], p=0.02). We found no associations between SNPs in the promoter region and TNF-α levels. In this study, innate cytokine production of TNF-α, IL-1β, IL-6 and IL-10 was not

  13. Evaluation of a low-cost electrostatic dust fall collector for indoor air endotoxin exposure assessment.

    Science.gov (United States)

    Noss, Ilka; Wouters, Inge M; Visser, Maaike; Heederik, Dick J J; Thorne, Peter S; Brunekreef, Bert; Doekes, Gert

    2008-09-01

    Exposure to endotoxin in home environments has become a key issue in asthma and allergy research. Most studies have analyzed floor or mattress dust endotoxin, but its validity as a proxy for airborne exposure is unknown, while active airborne dust sampling is not feasible in large-scale population studies because of logistic and financial limitations. We therefore developed and evaluated a simple passive airborne dust collection method for airborne endotoxin exposure assessment. We explored an electrostatic dust fall collector (EDC), consisting of a 42- by 29.6-cm-sized folder with four electrostatic cloths exposed to the air. The EDC was tested during two 14-day periods in seven nonfarm and nine farm homes and in farm stables. In parallel, active airborne dust sampling was performed with Harvard impactors and floor dust collected by vacuuming, using nylon sampling socks. The endotoxin levels could be measured in all EDC cloth extracts. The levels (in EU/m(2)) between EDCs used simultaneously or in different sampling periods in the same home correlated strongly (r > 0.8). EDC endotoxin also correlated moderately to strongly (r = 0.6 to 0.8) with the endotoxin measured by active airborne dust sampling and living room floor dust sampling and-in farm homes-with the endotoxin captured by the EDC in stables. In contrast, endotoxin levels measured by floor dust sampling showed only a poor correlation with the levels measured by active airborne dust sampling. We therefore conclude that measuring endotoxin levels with the EDC is a valid measure of average airborne endotoxin exposure, while reproducibility over time is at least equivalent to that of reservoir dust analyses.

  14. Evaluation of a Low-Cost Electrostatic Dust Fall Collector for Indoor Air Endotoxin Exposure Assessment▿

    Science.gov (United States)

    Noss, Ilka; Wouters, Inge M.; Visser, Maaike; Heederik, Dick J. J.; Thorne, Peter S.; Brunekreef, Bert; Doekes, Gert

    2008-01-01

    Exposure to endotoxin in home environments has become a key issue in asthma and allergy research. Most studies have analyzed floor or mattress dust endotoxin, but its validity as a proxy for airborne exposure is unknown, while active airborne dust sampling is not feasible in large-scale population studies because of logistic and financial limitations. We therefore developed and evaluated a simple passive airborne dust collection method for airborne endotoxin exposure assessment. We explored an electrostatic dust fall collector (EDC), consisting of a 42- by 29.6-cm-sized folder with four electrostatic cloths exposed to the air. The EDC was tested during two 14-day periods in seven nonfarm and nine farm homes and in farm stables. In parallel, active airborne dust sampling was performed with Harvard impactors and floor dust collected by vacuuming, using nylon sampling socks. The endotoxin levels could be measured in all EDC cloth extracts. The levels (in EU/m2) between EDCs used simultaneously or in different sampling periods in the same home correlated strongly (r > 0.8). EDC endotoxin also correlated moderately to strongly (r = 0.6 to 0.8) with the endotoxin measured by active airborne dust sampling and living room floor dust sampling and—in farm homes—with the endotoxin captured by the EDC in stables. In contrast, endotoxin levels measured by floor dust sampling showed only a poor correlation with the levels measured by active airborne dust sampling. We therefore conclude that measuring endotoxin levels with the EDC is a valid measure of average airborne endotoxin exposure, while reproducibility over time is at least equivalent to that of reservoir dust analyses. PMID:18676704

  15. An Exploratory Investigation of Endotoxin Levels in Novice Long Distance Triathletes, and the Effects of a Multi-Strain Probiotic/Prebiotic, Antioxidant Intervention.

    Science.gov (United States)

    Roberts, Justin D; Suckling, Craig A; Peedle, Georgia Y; Murphy, Joseph A; Dawkins, Tony G; Roberts, Michael G

    2016-11-17

    Gastrointestinal (GI) ischemia during exercise is associated with luminal permeability and increased systemic lipopolysaccharides (LPS). This study aimed to assess the impact of a multistrain pro/prebiotic/antioxidant intervention on endotoxin unit levels and GI permeability in recreational athletes. Thirty healthy participants (25 males, 5 females) were randomly assigned either a multistrain pro/prebiotic/antioxidant (LAB⁴ANTI; 30 billion CFU·day(-1) containing 10 billion CFU·day(-1)Lactobacillus acidophilus CUL-60 (NCIMB 30157), 10 billion CFU·day(-1)Lactobacillus acidophillus CUL-21 (NCIMB 30156), 9.5 billion CFU·day(-1)Bifidobacterium bifidum CUL-20 (NCIMB 30172) and 0.5 billion CFU·day(-1)Bifidobacterium animalis subspecies lactis CUL-34 (NCIMB 30153)/55.8 mg·day(-1) fructooligosaccharides/ 400 mg·day(-1) α-lipoic acid, 600 mg·day(-1)N-acetyl-carnitine); matched pro/prebiotic (LAB⁴) or placebo (PL) for 12 weeks preceding a long-distance triathlon. Plasma endotoxin units (via Limulus amebocyte lysate chromogenic quantification) and GI permeability (via 5 h urinary lactulose (L): mannitol (M) recovery) were assessed at baseline, pre-race and six days post-race. Endotoxin unit levels were not significantly different between groups at baseline (LAB⁴ANTI: 8.20 ± 1.60 pg·mL(-1); LAB⁴: 8.92 ± 1.20 pg·mL(-1); PL: 9.72 ± 2.42 pg·mL(-1)). The use of a 12-week LAB⁴ANTI intervention significantly reduced endotoxin units both pre-race (4.37 ± 0.51 pg·mL(-1)) and six days post-race (5.18 ± 0.57 pg·mL(-1); p = 0.03, ηp² = 0.35), but only six days post-race with LAB⁴ (5.01 ± 0.28 pg·mL(-1); p = 0.01, ηp² = 0.43). In contrast, endotoxin units remained unchanged with PL. L:M significantly increased from 0.01 ± 0.01 at baseline to 0.06 ± 0.01 with PL only (p = 0.004, ηp² = 0.51). Mean race times (h:min:s) were not statistically different between groups despite faster times with both pro/prebiotoic groups (LAB⁴ANTI: 13:17:07 ± 0

  16. An Exploratory Investigation of Endotoxin Levels in Novice Long Distance Triathletes, and the Effects of a Multi-Strain Probiotic/Prebiotic, Antioxidant Intervention

    Directory of Open Access Journals (Sweden)

    Justin D. Roberts

    2016-11-01

    Full Text Available Gastrointestinal (GI ischemia during exercise is associated with luminal permeability and increased systemic lipopolysaccharides (LPS. This study aimed to assess the impact of a multistrain pro/prebiotic/antioxidant intervention on endotoxin unit levels and GI permeability in recreational athletes. Thirty healthy participants (25 males, 5 females were randomly assigned either a multistrain pro/prebiotic/antioxidant (LAB4ANTI; 30 billion CFU·day−1 containing 10 billion CFU·day−1 Lactobacillus acidophilus CUL-60 (NCIMB 30157, 10 billion CFU·day−1 Lactobacillus acidophillus CUL-21 (NCIMB 30156, 9.5 billion CFU·day−1 Bifidobacterium bifidum CUL-20 (NCIMB 30172 and 0.5 billion CFU·day−1 Bifidobacterium animalis subspecies lactis CUL-34 (NCIMB 30153/55.8 mg·day−1 fructooligosaccharides/ 400 mg·day−1 α-lipoic acid, 600 mg·day−1 N-acetyl-carnitine; matched pro/prebiotic (LAB4 or placebo (PL for 12 weeks preceding a long-distance triathlon. Plasma endotoxin units (via Limulus amebocyte lysate chromogenic quantification and GI permeability (via 5 h urinary lactulose (L: mannitol (M recovery were assessed at baseline, pre-race and six days post-race. Endotoxin unit levels were not significantly different between groups at baseline (LAB4ANTI: 8.20 ± 1.60 pg·mL−1; LAB4: 8.92 ± 1.20 pg·mL−1; PL: 9.72 ± 2.42 pg·mL−1. The use of a 12-week LAB4ANTI intervention significantly reduced endotoxin units both pre-race (4.37 ± 0.51 pg·mL−1 and six days post-race (5.18 ± 0.57 pg·mL−1; p = 0.03, ηp2 = 0.35, but only six days post-race with LAB4 (5.01 ± 0.28 pg·mL−1; p = 0.01, ηp2 = 0.43. In contrast, endotoxin units remained unchanged with PL. L:M significantly increased from 0.01 ± 0.01 at baseline to 0.06 ± 0.01 with PL only (p = 0.004, ηp2 = 0.51. Mean race times (h:min:s were not statistically different between groups despite faster times with both pro/prebiotoic groups (LAB4ANTI: 13:17:07 ± 0:34:48; LAB4: 12

  17. Increased IgA responses to the LPS of commensal bacteria is associated with inflammation and activation of cell-mediated immunity in chronic fatigue syndrome.

    Science.gov (United States)

    Maes, Michael; Twisk, Frank N M; Kubera, Marta; Ringel, Karl; Leunis, Jean-Claude; Geffard, Michel

    2012-02-01

    Myalgic Encephalomyelitis/Chronic Fatigue Syndrome (ME/CFS) is accompanied by a) systemic IgA/IgM responses against the lipopolysaccharides (LPS) of commensal bacteria; b) inflammation, e.g. increased plasma interleukin-(IL)1 and tumor necrosis factor (TNF)α; and c) activation of cell-mediated immunity (CMI), as demonstrated by increased neopterin. To study the relationships between the IgA/IgM responses to the LPS of microbiota, inflammation, CMI and the symptoms of ME/CFS we measured the IgA/IgM responses to the LPS of 6 different enterobacteria, serum IL-1, TNFα, neopterin, and elastase in 128 patients with ME/CFS and chronic fatigue (CF). Severity of symptoms was assessed by the Fibromyalgia and Chronic Fatigue Syndrome (FF) Rating Scale. Serum IL-1, TNFα, neopterin and elastase are significantly higher in patients with ME/CFS than in CF patients. There are significant and positive associations between the IgA responses to LPS and serum IL-1, TNFα, neopterin and elastase. Patients with an abnormally high IgA response show increased serum IL-1, TNFα and neopterin levels, and higher ratings on irritable bowel syndrome (IBS) than subjects with a normal IgA response. Serum IL-1, TNFα and neopterin are significantly related to fatigue, a flu-like malaise, autonomic symptoms, neurocognitive disorders, sadness and irritability. The findings show that increased IgA responses to commensal bacteria in ME/CFS are associated with inflammation and CMI activation, which are associated with symptom severity. It is concluded that increased translocation of commensal bacteria may be responsible for the disease activity in some ME/CFS patients. Copyright © 2011 Elsevier B.V. All rights reserved.

  18. Development of methods for recovering endotoxins from surfaces and from air in production environment of injectable drugs.

    Science.gov (United States)

    Frucci, Fiammetta; Carducci, Annalaura; Davini, Giulia; Macchia, Marco; Bertini, Simone; Vecchiani, Sandra; Ceccanti, Stefano

    2017-08-17

    The aim of this work is to develop and validate methods for quantify endotoxins on surfaces and in the air of manufacturing environment of injectable drugs, in order to use them for evaluate the quality of the process and the risk for the products processed therein. The method for recovering endotoxins from surfaces was a direct method which provides sampling surfaces by swabbing and extraction of endotoxins from the swabs with an appropriate diluent; while the method for airborne endotoxins provides an air active sampling on a glass fiber filter (GFF) and endotoxins extraction with an appropriate diluent. Copyright © 2017, Parenteral Drug Association.

  19. Influence of the apical enlargement size on the endotoxin level reduction of dental root canals

    Directory of Open Access Journals (Sweden)

    Ariane Cassia Salustiano Marinho

    2012-12-01

    Full Text Available Gram-negative bacteria play an essential role in endodontic infections because they have virulence factors such as endotoxin. Due to its potential cytotoxic activity, special attention has been given to the removal/neutralization of this endotoxin in the root canal system. OBJECTIVE: The aim of this study was to evaluate the influence of the apical enlargement size (AES by using rotary instruments on the endotoxin level reduction of dental root canals. MATERIAL AND METHODS: Forty root canals of the mandibular premolar teeth were used. Escherichia coli endotoxin (055: B55 was inoculated into thirty root canals. Ten teeth served as the negative control group. After the incubation period, the first endotoxin samples were collected from the root canals with a sterile/apyrogenic paper point for the analysis of the endotoxin units (EU/mL present before instrumentation (S1. Specimen instrumentation was performed with the Mtwo® rotary system in the sequence 10/.04, 15/.05, 20/.06, 25/.06, 30/.05, 35/.04 and 40/.04. To monitor the effectiveness of increasing apical enlargement on endotoxin removal, the second endotoxin samples were collected from all the root canals after instrumentation with the following instruments: #25/.06- (S2; #30/.05- (S3; # 35/.04- (S4; and #40/.04- (S5. Limulus amebocyte lysate (LAL was used to quantify the levels of endotoxin. The results were statistically compared by using repeated measures of ANOVA with post hoc Tukey testing. RESULTS: Increasing levels of endotoxin removal was achieved by large sized apical enlargement: S2 (AES #25/.06- 89.2%, S3 (AES #30/.05- 95.9%, S4 (AES #35/.04- 97.8% and S5 (AES #40/.04- 98.2%. Substantial reduction of endotoxin content was obtained in S4 and S5 compared to S2 (p<0.05, however, the root canal preparation was not able to eliminate the endotoxin. CONCLUSIONS: Under the conditions of this study, it was concluded that the reduction of endotoxin levels of the dental root canals could be

  20. Punicalagin inhibits neuroinflammation in LPS-activated rat primary microglia.

    Science.gov (United States)

    Olajide, Olumayokun A; Kumar, Asit; Velagapudi, Ravikanth; Okorji, Uchechukwu P; Fiebich, Bernd L

    2014-09-01

    In this study, the effects of punicalagin on neuroinflammation in LPS-activated microglia were investigated. The ability of punicalagin to reduce the production of TNF-α, IL-6 and prostaglandin E2 was measured in culture medium using enzyme immunoassay. TNF-α and IL-6 gene expression in mouse hippocampal slices was measured with PCR. cyclooxygenase-2 and microsomal prostaglandin E synthase 1 protein and mRNA were evaluated with Western blotting and PCR, respectively. Further experiments to investigate effects of punicalagin on protein expressions of inflammatory targets were also determined with Western blotting. Pretreatment of rat primary microglia with punicalagin (5-40 μM) prior to LPS (10 ng/mL) stimulation produced a significant (p Punicalagin completely abolished TNF-α and IL-6 gene expression in LPS-stimulated hippocampal slices. Protein and mRNA expressions of cyclooxygenase-2 and microsomal prostaglandin E synthase 1 were also reduced by punicalagin pretreatment. Results show that punicalagin interferes with NF-κB signalling through attenuation of NF-κB-driven luciferase expression, as well as inhibition of IκB phosphorylation and nuclear translocation of p65 subunit in the microglia. These results suggest that punicalagin inhibits neuroinflammation in LPS-activated microglia through interference with NF-κB signalling, suggesting its potential as a nutritional preventive strategy in neurodegenerative disorders. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Blood purification for critical care medicine: endotoxin adsorption.

    Science.gov (United States)

    Kodama, M; Hanasawa, K; Tani, T

    1997-08-01

    Many kinds of blood purifying technologies have been applied to the treatment of critically ill patients since 1979 when plasma exchange with hollow-fiber membranes was developed. These technologies have been applied not only to the removal of toxic substances, but also to the treatment of objective diseases and the removal of the factors relating to the associated inflammation. This article summarizes these methods and their efficacies for critically ill patients, especially those with severe sepsis. Attempts have been made to remove endotoxin, the main cause of sepsis, from the circulation using polymyxin B immobilized fiber, charcoal hemoperfusion, and plasma or whole blood exchange. Attempts have also been made to remove proinflammatory cytokines, eicosanoides, and coagulative factors from the circulation in the human body. Continuous hemofiltration or hemodiafiltration is the representative technology. The efficacy of these methods has been established, but several issues remain unresolved. All methods of the treatment of severe sepsis are discussed with reference to treatment indications, efficacy, and outcome parameters. In particular, the clinical results of endotoxin removal with polymyxin B immobilized fiber are summarized in this article.

  2. Detection of microorganisms, endotoxins and aluminum in mobile dialysis services

    Directory of Open Access Journals (Sweden)

    Priscila Rodrigues de Jesus

    2017-11-01

    Full Text Available Hemodialysis water and dialysates are fundamental in the treatment of kidney disease. During just one hemodialysis session, 120 liters of dialysate are consumed. Thus, it is essential that the parameters of chemical and microbiological quality of the fluids used in dialysis systems are carefully observed. In this study, water samples were collected at 12 hospitals in the state of Rio de Janeiro. The samples were obtained at three points of fluid reservoirs: pre-, post-osmosis and dialysis solution. After collection, colony forming units (CFU, total coliforms and Escherichia coli 100 mL-1 were quantified. Later, isolated colonies and endotoxin content were identified by biochemical assays. Data about total aluminum levels per sample (mg L-1 were also obtained. Samples of all mobile dialysis services and points of collection were contaminated above the levels set out by national laws, in particular by Pseudomonas aeruginosa. Endotoxin levels were also above the recommended by current legislation (> 0.25 EU mL-1. Only three samples contained detectable levels of aluminum, which were found to be above the recommended values for the corresponding resolution (0.01 mg L-1. Finally, there were no observable amounts of total coliforms and E. coli 100 mL-1 sample. The data from this study are an important step forward in the standardization and control of chemical/microbiological quality of mobile dialysis services.

  3. Nonabsorbable Antibiotics Reduce Bacterial and Endotoxin Translocation in Hepatectomised Rats

    Directory of Open Access Journals (Sweden)

    S. K. Kakkos

    1997-01-01

    Full Text Available There is increasing evidence that septic complications, occurring after major hepatectomies, may be caused by gram negative bacteria, translocating from the gut. We investigated in rats, the effect of extended hepatectomy on the structure and morphology of the intestinal mucosa as well as on the translocation of intestinal bacteria and endotoxins. We also examined the effect of nonabsorbable antibiotics on reducing the intestinal flora and consequently the phenomenon of translocation by administering neomycin sulphate and cefazoline. Hepatectomy was found to increase translocation, while administration of nonabsorbable antibiotics decreased it significantly. In addition, hepatectomy increased the aerobic cecal bacterial population, which normalised in the group receiving antibiotics. Among the histological parameters evaluated, villus height demonstrated a significant reduction after hepatectomy, while the number of villi per cm and the number of mitoses per crypt, remained unchanged. Our results indicate that administration of nonabsorbable antibiotics presents a positive effect on bacterial and endotoxin translocation after extended hepatectomy, and this may be related to reduction of colonic bacterial load as an intraluminal effect of antibiotics.

  4. Measurement of endotoxins in bioaerosols at workplace: a critical review of literature and a standardization issue.

    Science.gov (United States)

    Duquenne, Philippe; Marchand, Geneviève; Duchaine, Caroline

    2013-03-01

    Endotoxins are lipopolysaccharides found in the outer membrane of most Gram-negative bacteria and cyanobacteria. Worker exposure to endotoxins has been shown in a number of work situations and is associated with both respiratory and systemic pathologies. The lack of an occupational exposure limit is mainly due to the absence of a standard protocol at the international level for sampling and analyzing airborne endotoxins. The bibliographic review in this article takes an exhaustive look at the current knowledge on measuring airborne endotoxins. It shows that, despite several reference documents at the international level, the methods used to measure endotoxin exposure differ considerably from one laboratory to another. Standardization is necessary to reduce interlaboratory variability and, ultimately, to improve the use of interstudy data. The bibliographic review presents the current status of standardization for airborne endotoxin measurement methods in the workplace and summarizes areas for further research. This article is both a reference document for all operators wishing to use such methods and a working document to build international consensus around the measurement of airborne endotoxins.

  5. Quantification of the Influence of Endotoxins on the Mechanics of Adult and Neonatal Red Blood Cells.

    Science.gov (United States)

    Ito, Hiroaki; Kuss, Navina; Rapp, Bastian E; Ichikawa, Masatoshi; Gutsmann, Thomas; Brandenburg, Klaus; Pöschl, Johannes M B; Tanaka, Motomu

    2015-06-25

    In this study, we physically modeled the influence of endotoxin-induced sepsis symptoms on human red blood cells (RBCs) by quantifying the impact of endotoxins on the cell mechanics by the analysis of Fourier-transformed mean square amplitude of shape fluctuation, called flicker spectroscopy. With the aid of a microfluidic diffusion chamber, we noninvasively determined principal mechanical parameters of human RBCs in the absence and presence of endotoxins for individual RBCs for the first time. Because of the elongation of saccharide chain length of endotoxins, we found an increase in the morphological transition from discocytes to echinocytes, and monotonic changes in the mechanical parameters. Since septic shocks often cause lethal risks of neonates, we measured the mechanical parameters of neonatal RBCs, and compared them to those of adult RBCs. The quantitative comparison reveals that neonatal RBCs are more susceptible to the effect of endotoxins than adult RBCs. Furthermore, coincubation with the antiseptic peptide P19-2.5 (Aspidasept) with endotoxin results in a slight suppression of the impact of the endotoxin. The strategy proposed in our study can potentially be applied for the quantitative diagnosis of RBCs based on mechanical readouts.

  6. In silico designed nanoMIP based optical sensor for endotoxins monitoring.

    Science.gov (United States)

    Abdin, M J; Altintas, Z; Tothill, I E

    2015-05-15

    Molecular modelling was used to select specific monomers suitable for the design of molecularly imprinted polymers (MIPs) with high affinity towards endotoxins. MIPs were synthesised using solid-phase photopolymerisation with endotoxins from Escherichia coli 0111:B4 as the template. This technique also allowed the endotoxin template to be reused successfully. Particle size of ~190-220 nm was achieved with low polydispersity index, which confirms the quality of the produced MIPs. For the development of the optical sensor, SPR-2 biosensor system was used by functionalising the gold sensor chip with the MIP nanoparticles using EDC/NHS coupling procedure. The affinity based-endotoxin assay can detect endotoxins in the concentration range of 15.6-500 ng mL(-1). MIP surfaces were regenerated showing stability of the method for subsequent analysis and dissociation constants were calculated as 3.24-5.24×10(-8) M. The developed SPR sensor with the novel endotoxins nanoMIP showed the potential of the technology for endotoxins capture, detection and risk management and also the importance of computational modelling to design the artificial affinity ligands. Copyright © 2014 Elsevier B.V. All rights reserved.

  7. Biologically Active Endotoxins from Salmonella Mutants Deficient in O- and R-Polysaccharides and Heptose

    Science.gov (United States)

    Kim, Yoon Berm; Watson, Dennis W.

    1967-01-01

    Well-characterized Salmonella mutants formerly used in biosynthetic studies of lipopolysaccharides were used to study the toxic portion of the complex endotoxin. Endotoxins prepared from wild types and their mutants were tested for their biological activities, including pyrogenicity, lethality, and immunogenicity. There was little difference either in the endotoxin yields or in the toxicities between endotoxins from the wild-type and O-antigen deficient mutants. Endotoxin containing mostly lipid A and keto-deoxyoctonate (KDO) prepared from the mutant deficient in both O- and R-antigens and the backbone sugar, heptose, was biologically active. Possibly because of the difference in solubility in water, the yield of endotoxin from the heptoseless mutant was about 10% of the wild type. There was complete reciprocal cross-immunity between all endotoxins tested. These observations suggest that the common toxic moiety is not present in the O- and R-polysaccharides or the backbone sugar heptose, but rather is associated with the lipid portion of the molecule which includes mostly lipid A and KDO. PMID:4862190

  8. Inhibition of endotoxin effects on cultured human middle ear epithelium by bactericidal permeability-increasing protein.

    Science.gov (United States)

    Nell, M J; Albers-Op 't Hof, B M; Koerten, H K; Grote, J J

    2000-09-01

    Endotoxin can induce morphologic changes to middle ear epithelium, which can disturb the mucociliary clearance system (MCS) and lead to otitis media with effusion (OME). The bactericidal/permeability-increasing (BPI) protein is a major component of neutrophil granules and binds with high affinity to endotoxin. In this study, the capacity to inhibit the effects of endotoxin by rBPI21, a recombinant amino-terminal analog derived from BPI, was investigated on cultured human middle ear epithelium using light microscopy and scanning- and transmission electron microscopy. Human middle ear epithelium was air-exposed cultured on a collagenous underlayer with different additions of endotoxin and rBPI21 to the culture medium. The tissue specimens were inspected after 4 weeks for the number of ciliated and secretory cells, thickness of the mucosal layer, and cell size. The morphologic changes induced by endotoxin were increased thickness of the mucosal layer and increased number of secretory cells. These changes were significantly diminished or even absent when endotoxin was added with rBPI21 to the culture medium. rBPI21 can inhibit morphologic changes in the middle ear epithelium due to endotoxin. Hence, the authors believe that rBPI21 can be a new therapeutic agent in the treatment of OME.

  9. Silybin attenuates LPS-induced lung injury in mice by inhibiting NF-κB signaling and NLRP3 activation.

    Science.gov (United States)

    Zhang, Bo; Wang, Bing; Cao, Shuhua; Wang, Yongqiang; Wu, Di

    2017-05-01

    Silybin is one of the main flavonoids produced by milk thistle, which has been used in the treatment of liver diseases. In this study, we examined the protective effects and possible mechanisms of action of silybin in lipopolysaccharide (LPS)‑induced lung injury and inflammation. Pre-treatment of mice with silybin significantly inhibited LPS-induced airway inflammatory cell recruitment, including macrophages, T cells and neutrophils. The production of cytokines, such as interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in bronchoalveolar fluid and serum was also decreased following treatment with silybin. Elevated cytokine mRNA levels induced by LPS in lung tissue were all suppressed by silybin and lung histological alterations were also improved. In addition, experiments using cells indicated that silybin significantly decreased the mRNA levels and secretion of IL-1β and TNF-α in THP-1 cells. Moreover, the mechanisms responsible for these effects were attributed to the inhibitory effect of silybin on nuclear factor-κB (NF-κB) signaling and NLR family pyrin domain containing 3 (NLRP3) inflammasome activation. The data form our study thus support the utility of silybin as a potential medicine for the treatment of acute lung injury‑associated inflammation and pathological changes. Silybin exerts protective effects against lung injury by regulating NF-κB signaling and the NLRP3 inflammasome activation.

  10. Racial differences in the association of CD14 polymorphisms with serum total IgE levels and allergen skin test reactivity

    Directory of Open Access Journals (Sweden)

    Wang ZY

    2013-06-01

    disease. For subjects with CD14 1188 G alleles, the association with atopic disease was stronger in Caucasians compared to African Americans.Keywords: total serum immunoglobulin E, IgE, skin prick test, SPT, CD14-159T, single nucleotide polymorphism, SNP, lipopolysaccharide, LPS, endotoxin

  11. House Dust Endotoxin Levels Are Associated with Adult Asthma in a U.S. Farming Population.

    Science.gov (United States)

    Carnes, Megan Ulmer; Hoppin, Jane A; Metwali, Nervana; Wyss, Annah B; Hankinson, John L; O'Connell, Elizabeth Long; Richards, Marie; Long, Stuart; Freeman, Laura E Beane; Sandler, Dale P; Henneberger, Paul K; Barker-Cummings, Christie; Umbach, David M; Thorne, Peter S; London, Stephanie J

    2017-03-01

    Endotoxin initiates a proinflammatory response from the innate immune system. Studies in children suggest that endotoxin exposure from house dust may be an important risk factor for asthma, but few studies have been conducted in adult populations. To investigate the association of house dust endotoxin levels with asthma and related phenotypes (wheeze, atopy, and pulmonary function) in a large U.S. farming population. Dust was collected from the bedrooms (n = 2,485) of participants enrolled in a case-control study of current asthma (927 cases) nested within the Agricultural Health Study. Dust endotoxin was measured by Limulus amebocyte lysate assay. Outcomes were measured by questionnaire, spirometry, and blood draw. We evaluated associations using linear and logistic regression. Endotoxin was significantly associated with current asthma (odds ratio [OR], 1.30; 95% confidence interval [CI], 1.14-1.47), and this relationship was modified by early-life farm exposure (born on a farm: OR, 1.18; 95% CI, 1.02-1.37; not born on a farm: OR, 1.67; 95% CI, 1.26-2.20; Interaction P = 0.05). Significant positive associations were seen with both atopic and nonatopic asthma. Endotoxin was not related to either atopy or wheeze. Higher endotoxin was related to lower FEV1/FVC in asthma cases only (Interaction P = 0.01). For asthma, there was suggestive evidence of a gene-by-environment interaction for the CD14 variant rs2569190 (Interaction P = 0.16) but not for the TLR4 variants rs4986790 and rs4986791. House dust endotoxin was associated with current atopic and nonatopic asthma in a U.S. farming population. The degree of the association with asthma depended on early-life farm exposures. Furthermore, endotoxin was associated with lower pulmonary function in patients with asthma.

  12. The molecular adsorption-type endotoxin detection system using immobilized ɛ-polylysine

    Science.gov (United States)

    Ooe, Katsutoshi; Tsuji, Akihito; Nishishita, Naoki; Hirano, Yoshiaki

    2007-12-01

    Hemodialysis for chronic renal failure is the most popular treatment method with artificial organs. However, hemodialysis patients must continue the treatment throughout their life, the results of long term extracorporeal dialysis, those patients develop the various complications and diseases, for example, dialysis amyloidosis etc. Dialysis amyloidosis is one of the refractory complications, and endotoxin is thought to be the most likely cause of it, recently. Endotoxin is one of the major cell wall components of gram-negative bacteria, and it has various biological activities. In addition, it is known that a mount of endotoxin exists in living environment, and medicine is often contaminated with endotoxin. When contaminated dialyzing fluids are used to hemodialysis, above-mentioned dialysis amyloidosis is developed. Therefore, it is important that the detection and removal of endotoxin from dialyzing fluids. Until now, the measurement methods using Limulus Amebosyte Lysate (LAL) reagent were carried out as the tests for the presence of endtoxin. However, these methods include several different varieties of measurement techniques. The following are examples of them, gelatinization method, turbidimetric assay method, colorimetric assay method and fluoroscopic method. However, these techniques needed 30-60 minutes for the measurement. From these facts, they are not able to use as a "real-time endotoxin detector". The detection of endotoxin has needed to carry out immediately, for that reason, a new detection method is desired. In this research, we focused attention to adsorption reaction between ɛ-polylysine and endotoxin. ɛ-polylysine has the structure of straight chain molecule composed by 25-30 residues made by lysine, and it is used as an antimicrobial agent, moreover, cellulose beads with immobilized ɛ-polylysine is used as the barrier filter for endotoxin removal. The endotoxin is adsorbed to immobilized ɛ-polylysine, as the result of this reaction, the

  13. Effect of plasmapheresis on the immune system in endotoxin-induced sepsis

    DEFF Research Database (Denmark)

    Toft, P; Schmidt, R; Broechner, A C

    2008-01-01

    BACKGROUND: It has been proposed that plasmapheresis is most effective when applied early in Gram-negative sepsis. We therefore studied the effect of early plasmapheresis on immunity in experimental Escherichia coli endotoxin-induced sepsis. METHODS: 20 pigs received 30 microg/kg of E. coli...... endotoxin. 40 min later, half of the pigs were treated with plasmapheresis which lasted 4 h. The adhesion molecules, the oxidative burst, the number of neutrophils in blood and lungs, and cytokines were measured. RESULTS: Infusion of endotoxin was associated with activation of adhesion molecules increased...... infusion, it only temporarily attenuated a part of the activated cell-mediated immunity....

  14. Human Milk Shows Immunological Advantages Over Organic Milk Samples For Infants in the Presence of Lipopolysaccharide (LPS in 3D Energy Maps Using an Organic Nanobiomimetic Memristor/Memcapacitor

    Directory of Open Access Journals (Sweden)

    S-H. DUH

    2016-08-01

    Full Text Available Human milk is well known for its immunological advantages of protection and support for healthy early childhood cognitive development and prevention of chronic diseases over cow milk for infants. However, little is known about how the immunological advantages are linked to reduce Pathological High Frequency Oscillation (pHFO regarding neural synapse net energy outcomes when lipopolysaccharide (LPS attacks at a clinical concentration range compared with that in cow milk in a 3D energy map. We developed a nanostructure biomimetic memristor/memcapacitor device with a dual function of chronoamperometric (CA sensing/voltage sensing for the direct quantitative evaluation of immunological advantages between human milk and organic cow milk for infants in the presence of wide LPS concentration ranges; those ranges were between 5.0 pg/mL to 500 ng/mL and from 50 ng/mL to 1 µg/mL for both a CA and a voltage method, respectively. The Detection of Limit (DOL results are as follows: 3.73×10-18 g LPS vs. 1.2×10-16 g LPS in 40 µL milk samples using the 3.11×10-7cm3 voltage sensor and the 0.031cm2 CA sensor, respectively, under antibody-free and reagent-free conditions. The 3D energy map results show that cow milk is ten-times more prone to E. Coli attack, and the positive link was revealed that Pathological High Frequency Oscillation (pHFO formations occurred over the studied LPS concentration range from 50 ng/mL up to 1000 ng/mL from Rapid Eye Movement (REM sleep frequency, fast gamma frequency to Sharp Wave-Ripple Complexes (SPW- R frequency. There had no pHFO with human milk samples at Slow Wave Sleeping (SWS, REM and SPW- R frequencies. The microbiota in the human milk samples successfully overcame the endotoxin attack from E. coli bacteria, however the pHFO only occurred at fast gamma frequency linked with the LPS level ≥ 500 ng/mL. Organic milk samples show an order of magnitude lower synapse energy density compared with human milk at SWS for with

  15. Progesterone is essential for protecting against LPS-induced pregnancy loss. LIF as a potential mediator of the anti-inflammatory effect of progesterone.

    Directory of Open Access Journals (Sweden)

    Julieta Aisemberg

    Full Text Available Lipopolysaccharide (LPS administration to mice on day 7 of gestation led to 100% embryonic resorption after 24 h. In this model, nitric oxide is fundamental for the resorption process. Progesterone may be responsible, at least in part, for a Th2 switch in the feto-maternal interface, inducing active immune tolerance against fetal antigens. Th2 cells promote the development of T cells, producing leukemia inhibitory factor (LIF, which seems to be important due to its immunomodulatory action during early pregnancy. Our aim was to evaluate the involvement of progesterone in the mechanism of LPS-induced embryonic resorption, and whether LIF can mediate hormonal action. Using in vivo and in vitro models, we provide evidence that circulating progesterone is an important component of the process by which infection causes embryonic resorption in mice. Also, LIF seems to be a mediator of the progesterone effect under inflammatory conditions. We found that serum progesterone fell to very low levels after 24 h of LPS exposure. Moreover, progesterone supplementation prevented embryonic resorption and LPS-induced increase of uterine nitric oxide levels in vivo. Results show that LPS diminished the expression of the nuclear progesterone receptor in the uterus after 6 and 12 h of treatment. We investigated the expression of LIF in uterine tissue from pregnant mice and found that progesterone up-regulates LIF mRNA expression in vitro. We observed that LIF was able to modulate the levels of nitric oxide induced by LPS in vitro, suggesting that it could be a potential mediator of the inflammatory action of progesterone. Our observations support the view that progesterone plays a critical role in a successful pregnancy as an anti-inflammatory agent, and that it could have possible therapeutic applications in the prevention of early reproductive failure associated with inflammatory disorders.

  16. DMPD: LPS, TLR4 and infectious disease diversity. [Dynamic Macrophage Pathway CSML Database

    Lifescience Database Archive (English)

    Full Text Available 15608698 LPS, TLR4 and infectious disease diversity. Miller SI, Ernst RK, Bader MW.... Nat Rev Microbiol. 2005 Jan;3(1):36-46. (.png) (.svg) (.html) (.csml) Show LPS, TLR4 and infectious disease... diversity. PubmedID 15608698 Title LPS, TLR4 and infectious disease diversity. Authors Miller SI, Ernst RK,

  17. On the role and fate of LPS-dephosphorylating activity in the rat liver

    NARCIS (Netherlands)

    Tuin, A; Huizinga -van der Vlag, Ali; van Loenen - Weemaes, Anne-miek; Meijer, DKF; Poelstra, K

    Gut-derived lipopolysaccharide (LPS) plays a role in the pathogenesis of liver diseases like fibrosis. The enzyme alkaline phosphatase (AP) is present in, among others, the intestinal wall and liver and has been previously shown to dephosphorylate LPS. Therefore, we investigated the effect of LPS on

  18. DMPD: The Lps locus: genetic regulation of host responses to bacteriallipopolysaccharide. [Dynamic Macrophage Pathway CSML Database

    Lifescience Database Archive (English)

    Full Text Available 10669111 The Lps locus: genetic regulation of host responses to bacteriallipopolysa...ccharide. Qureshi ST, Gros P, Malo D. Inflamm Res. 1999 Dec;48(12):613-20. (.png) (.svg) (.html) (.csml) Show The Lps locus: genetic... regulation of host responses to bacteriallipopolysaccharide. PubmedID 10669111 Title The Lps locus: genetic

  19. DMPD: LPS-binding proteins and receptors. [Dynamic Macrophage Pathway CSML Database

    Lifescience Database Archive (English)

    Full Text Available 9665271 LPS-binding proteins and receptors. Fenton MJ, Golenbock DT. J Leukoc Biol.... 1998 Jul;64(1):25-32. (.png) (.svg) (.html) (.csml) Show LPS-binding proteins and receptors. PubmedID 9665271 Title LPS-binding prot...eins and receptors. Authors Fenton MJ, Golenbock DT. Publication J Leukoc Biol. 199

  20. DMPD: LPS induction of gene expression in human monocytes. [Dynamic Macrophage Pathway CSML Database

    Lifescience Database Archive (English)

    Full Text Available 11257452 LPS induction of gene expression in human monocytes. Guha M, Mackman N. Ce...ll Signal. 2001 Feb;13(2):85-94. (.png) (.svg) (.html) (.csml) Show LPS induction of gene expression in human... monocytes. PubmedID 11257452 Title LPS induction of gene expression in human monocytes. Authors Guha M, Ma

  1. Comparison of result judgment algorithm of test for interfering factors in the bacterial endotoxins test among Chinese, Japanese, European, American, and Indian pharmacopeias.

    Science.gov (United States)

    Pei, Yusheng; Cai, Tong; Gao, Hua; Tan, Dejiang; Zhang, Yuchen; Zhang, Guolai

    2014-01-01

    The bacterial endotoxins test (BET) is a method used to detect or quantify endotoxins (lipo-polysaccharide, LPS) and is widely used in the quality control of parenteral medicines/vaccines and clinical dialysis fluid. It is also used in the diagnosis of endotoxemia and in detection of environment air quality control. Although BET has been adopted by most pharmacopoeias, result judgment algorithms (RJAs) of the test for interfering factors in the BET still differ between certain pharmacopoeias. We have evaluated RJAs of the test for interfering factors for the revision of BET described in the Chinese Pharmacopoeia 2010 (CHP2010). Original data from 1 748 samples were judged by RJAs of the Chinese Pharmacopoeia 2010, the Japanese Pharmacopoeia 2011 (JP2011), the European Pharmacopoeia 7.0 (EP7.0), the United States Pharmacopoeia 36 (USP36), and the Indian Pharmacopoeia 2010 (IP2010), respectively. A SAS software package was used in the statistical analysis. The results using CHP2010 and USP36, JP2011, EP7.0, and IP2010 had no significant difference (P = 0.7740). The results using CHP2010 of 1 748 samples showed that 132 samples (7.6%) required an additional step; nevertheless there was no such requirement when using the other pharmacopeias. The kappa value of two RJAs (CHP2010 and EP7.0) was 0.6900 (0.6297-0.7504) indicating that the CHP2010 and other pharmacopoeias have good consistency. The results using CHP2010 and USP36, JP2011, EP7.0, and IP2010 have different characteristics. CHP2010 method shows a good performance in Specificity, mistake diagnostic rate, agreement rate, predictive value for suspicious rate, and predictive value for passed rate. The CHP2010 method only had disadvantages in sensitivity compared with other pharmacopeias. We suggest that the Chinese pharmacopoeia interference test be revised in accordance with the USP36, JP2011, EP7.0, and IP2010 judgment model.

  2. Ambient endotoxin in PM10and association with inflammatory activity, air pollutants, and meteorology, in Chitwan, Nepal.

    Science.gov (United States)

    Mahapatra, Parth Sarathi; Jain, Sumeet; Shrestha, Sujan; Senapati, Shantibhusan; Puppala, Siva Praveen

    2018-03-15

    Endotoxin associated with ambient PM (particulate matter) has been linked to adverse respiratory symptoms, but there have been few studies of ambient endotoxin and its association with co-pollutants and inflammation. Our aim was to measure endotoxin associated with ambient PM 10 (particulate matter with aerodynamic diametermeteorology, co-pollutants, and inflammatory activity. PM 10 concentrations were recorded and filter paper samples were collected using E-samplers; PM 1, PM 2.5 , black carbon (BC), methane (CH 4 ), and carbon monoxide (CO) were also measured. The Limulus amebocyte lysate (LAL) assay was used for endotoxin quantification and the nuclear factor kappa B (NFκB) activation assay to assess inflammatory activity. The mean concentration of PM 10 at the different locations ranged from 136 to 189μg/m 3 , and of endotoxin from 0.29 to 0.53EU/m 3 . Pollutant presence was positively correlated with endotoxin. Apart from relative humidity, meteorological variations had no significant impact on endotoxin concentration. NF-κB activity was negatively correlated with endotoxin concentration. To the best of our knowledge, this study provides the first measurements of ambient endotoxin associated with PM 10 in Nepal. Endotoxin and co-pollutants were positively associated indicating a similar source. Endotoxin was negatively correlated with inflammatory activity as a result of a time-limited forest fire event during the sampling period. Studies of co-pollutants suggested that the higher levels of endotoxin related to biomass burning were accompanied by increased levels of anti-inflammatory agents, which suppressed the endotoxin inflammatory effect. Copyright © 2017. Published by Elsevier B.V.

  3. Air-sampled Filter Analysis for Endotoxins and DNA Content.

    Science.gov (United States)

    Lang-Yona, Naama; Mazar, Yinon; Pardo, Michal; Rudich, Yinon

    2016-03-07

    Outdoor aerosol research commonly uses particulate matter sampled on filters. This procedure enables various characterizations of the collected particles to be performed in parallel. The purpose of the method presented here is to obtain a highly accurate and reliable analysis of the endotoxin and DNA content of bio-aerosols extracted from filters. The extraction of high molecular weight organic molecules, such as lipopolysaccharides, from sampled filters involves shaking the sample in a pyrogen-free water-based medium. The subsequent analysis is based on an enzymatic reaction that can be detected using a turbidimetric measurement. As a result of the high organic content on the sampled filters, the extraction of DNA from the samples is performed using a commercial DNA extraction kit that was originally designed for soils and modified to improve the DNA yield. The detection and quantification of specific microbial species using quantitative polymerase chain reaction (q-PCR) analysis are described and compared with other available methods.

  4. Ginger extract inhibits LPS induced macrophage activation and function

    Directory of Open Access Journals (Sweden)

    Bruch David

    2008-01-01

    Full Text Available Abstract Background Macrophages play a dual role in host defence. They act as the first line of defence by mounting an inflammatory response to antigen exposure and also act as antigen presenting cells and initiate the adaptive immune response. They are also the primary infiltrating cells at the site of inflammation. Inhibition of macrophage activation is one of the possible approaches towards modulating inflammation. Both conventional and alternative approaches are being studied in this regard. Ginger, an herbal product with broad anti inflammatory actions, is used as an alternative medicine in a number of inflammatory conditions like rheumatic disorders. In the present study we examined the effect of ginger extract on macrophage activation in the presence of LPS stimulation. Methods Murine peritoneal macrophages were stimulated by LPS in presence or absence of ginger extract and production of proinflammatory cytokines and chemokines were observed. We also studied the effect of ginger extract on the LPS induced expression of MHC II, B7.1, B7.2 and CD40 molecules. We also studied the antigen presenting function of ginger extract treated macrophages by primary mixed lymphocyte reaction. Results We observed that ginger extract inhibited IL-12, TNF-α, IL-1β (pro inflammatory cytokines and RANTES, MCP-1 (pro inflammatory chemokines production in LPS stimulated macrophages. Ginger extract also down regulated the expression of B7.1, B7.2 and MHC class II molecules. In addition ginger extract negatively affected the antigen presenting function of macrophages and we observed a significant reduction in T cell proliferation in response to allostimulation, when ginger extract treated macrophages were used as APCs. A significant decrease in IFN-γ and IL-2 production by T cells in response to allostimulation was also observed. Conclusion In conclusion ginger extract inhibits macrophage activation and APC function and indirectly inhibits T cell activation.

  5. Experimental study of 『PERSICAE SEMEN』 on the blood injected by Endotoxin in rats

    Directory of Open Access Journals (Sweden)

    Chang-Keun

    2005-06-01

    Full Text Available This study was performed to investigate the effects of 「Persicae Semen」(PS on the blood injected by Endotoxin in rats. The blood was induced by Endotoxin injection into the caudal vein of rats and PS group taken a measurement of RBC, Hb, Hct, Platelet, WBC, ESR, CRP. The results were obtained as follows: 1. RBC, Hb, Hct, Platelet, WBC were increased with statistical significance at PS group as compared with those of the control group. 2. ESR, CRP were decreased with statistical significance at PS group as compared with those of the control group. It is concluded that PS group has significant effects on the blood injected by Endotoxin in rats. Therefore, PS group seems to be applicable to the diseases related to Endotoxin in clinics.

  6. The emerging role of microRNA in regulation of endotoxin tolerance.

    LENUS (Irish Health Repository)

    Quinn, Edel M

    2012-05-01

    Endotoxin tolerance is a phenomenon where cells show reduced responsiveness toward repeated endotoxin stimulation. Regulation of tolerance occurs at multiple levels of the cell signaling cascade, and many of these levels are potentially regulated by miRNA, which are a class of small RNA that bind to mRNA to down-regulate gene expression at the post-transcriptional level. Roles have been identified for miR-146a, miR-221, miR-579, miR-125b, miR-155, let-7e, and miR-98 in regulating the TLR4 signaling pathway during the development of endotoxin tolerance at receptor, signaling pathway, and gene transcription and translational levels. miRNA represent exciting, new potential targets in attempts to exogenously modulate development of endotoxin tolerance.

  7. Effect of saccharides on growth, sporulation rate and δ-endotoxin ...

    African Journals Online (AJOL)

    sucrose, lactose and maltose) as carbon sources on growth and sporulation rate of Bacillus thuringiensis MPK13 as well as δ-endotoxin production was investigated using 400 ml shake flasks culture. Substantially high growth and sporulation ...

  8. Isolation and sequencing of the cryIC-like delta endotoxin gene from ...

    African Journals Online (AJOL)

    Isolation and sequencing of the cryIC-like delta endotoxin gene from Egyptian strains of Bacillus thuringiensis toxic against Lepidoptera. ME Elkashef, AM Heikal, MEDM Solliman, MI Kobeasy, HA El-Shemy ...

  9. The Role of Bacterial Endotoxins in Occupational Diseases Caused by Inhaling Vegetable Dusts

    Science.gov (United States)

    Pernis, B.; Vigliani, E. C.; Cavagna, C.; Finulli, M.

    1961-01-01

    A large group of occupational diseases connected with the inhalation of various vegetable dusts, especially in the textile industry, have certain main symptoms in common such as fever, coughing, dyspnoea, and general malaise. In most cases the symptoms are more prominent on Mondays or on resuming work after one or more days of interruption. The symptomatology of these diseases and the Monday effect leads to the hypothesis that they are due to the inhalation of the endotoxins of gram-negative bacteria that contaminate the various vegetable materials, the Monday effect being connected with the phenomenon of tolerance to the endotoxins. Support for this view came from the demonstration of the constant presence of endotoxins in cotton dusts in textile mills and from the study of the effects of the inhalation of purified endotoxins in rabbits and man. Images PMID:13734460

  10. Prematurity does not markedly affect intestinal sensitivity to endotoxins and feeding in pigs.

    Science.gov (United States)

    Bering, Stine B; Bai, Shiping; Zhang, Keying; Sangild, Per T

    2012-08-01

    Preterm neonates show enhanced sensitivity to nutrient maldigestion and bacteria-mediated gut inflammatory disorders, such as necrotising enterocolitis (NEC). We hypothesised that preterm birth increases the sensitivity of intestinal nutrient absorption to endotoxins and that feeding after birth reduces this response. Hence, we investigated the postnatal development of nutrient digestive and absorptive capacity in the preterm and term pig intestine, and its responsiveness to endotoxins. Pigs were delivered by caesarean section at preterm (n 20) or term (n 17) gestation, and the small intestine was collected at birth or after 2 d of colostrum feeding, followed by ex vivo stimulation with lipopolysaccharide endotoxins and mixed gut contents collected from pigs with NEC. Brush border enzyme activities were reduced in newborn preterm v. term pigs (39-45 % reduction, P endotoxins could be less important, at least in pigs.

  11. Plasma endotoxin activity in Eastern grey kangaroos (Macropus giganteus) with lumpy jaw disease

    National Research Council Canada - National Science Library

    SOTOHIRA, Yukari; SUZUKI, Kazuyuki; OTSUKA, Marina; TSUCHIYA, Masakazu; SHIMAMORI, Toshio; NISHI, Yasunobu; TSUKANO, Kenji; ASAKAWA, Mitsuhiko

    2017-01-01

    ...)”, is one of the most significant causes of illness and death in captive macropods. The aim of the present study was to evaluate the relationship between the severity of LJD and plasma endotoxin activity in kangaroos...

  12. Transepithelial activation of human leukocytes by probiotics and commensal bacteria: role of Enterobacteriaceae-type endotoxin

    DEFF Research Database (Denmark)

    Bäuerlein, A.; Ackermann, S.; Parlesak, Alexandr

    2009-01-01

    The goal of the current study was to clarify whether commercially available probiotics induce greater trans-epithelial activation of human leukocytes than do commensal, food-derived and pathogenic bacteria and to identify the compounds responsible for this activation. Eleven different bacterial...... strains, and some of their pathogen-associated molecular patterns, were incubated apically on a confluent layer of intestinal epithelial cells (Caco-2), which were basolaterally co-cultured with human mononuclear leukocytes. Only Gram-negative bacteria having Enterobacteriaceae-type endotoxin (commensal...... fermenters (Lactobacillus spp.) did not stimulate leukocytes transepithelially. Endotoxin from E. coli and Salmonella enterica Typhimurium induced comparable transepithelial stimulation of leukocytes, but not endotoxin from B. vulgatus or lipoteichoic acid from E. faecalis. Endotoxin-binding agents...

  13. Transepithelial activation of human leukocytes by probiotics and commensal bacteria: Role of Enterobacteriaceae-type endotoxin

    DEFF Research Database (Denmark)

    Baeuerlein, Annette; Ackermann, Stefanie; Parlesak, Alexandr

    2009-01-01

    The goal of the current study was to clarify whether commercially available probiotics induce greater trans-epithelial activation of human leukocytes than do commensal, food-derived and pathogenic bacteria and to identify the compounds responsible for this activation. Eleven different bacterial...... strains, and some of their pathogen-associated molecular patterns, were incubated apically on a confluent layer of intestinal epithelial cells (Caco-2), which were basolaterally co-cultured with human mononuclear leukocytes. Only Gram-negative bacteria having Enterobacteriaceae-type endotoxin (commensal...... fermenters (Lactobacillus spp.) did not stimulate leukocytes transepithelially. Endotoxin from E. coli and Salmonella enterica Typhimurium induced comparable transepithelial stimulation of leukocytes, but not endotoxin from B. vulgatus or lipoteichoic acid from E. faecalis. Endotoxin-binding agents...

  14. Alfalfa pellet-induced subacute ruminal acidosis in dairy cows increases bacterial endotoxin in the rumen without causing inflammation.

    Science.gov (United States)

    Khafipour, E; Krause, D O; Plaizier, J C

    2009-04-01

    A study was conducted to determine if subacute ruminal acidosis (SARA) induced by feeding alfalfa pellets results in increases in free bacterial lipopolysaccharide (LPS) in rumen fluid and peripheral blood, and acute phase proteins in plasma, and to determine the effect of alfalfa pellet-induced SARA on feed intake, rumen fermentation characteristics, milk production and composition, and blood metabolites. Eight lactating Holstein cows, 4 of which were ruminally cannulated, were used in a 6-wk experiment and were fed once daily at 0900 h. During wk 1, cows received a diet containing 50% of DM as concentrate and 50% of DM chopped alfalfa hay. Between wk 2 and wk 6, alfalfa hay was gradually replaced with alfalfa pellets at the rate of 8% per week to reduce rumen pH. Rumen pH was monitored continuously in the ruminally cannulated cows using indwelling pH probes. Rumen fluid and peripheral blood were sampled 15 min before feed delivery and at 6 h after feed delivery. Based on adopted threshold of SARA of at least 180 min/d below pH 5.6, SARA was induced from wk 3 onwards. Replacing 40% of alfalfa hay with alfalfa pellets quadratically increased the DMI from 18.1 kg/d in wk 1 to 23.4 kg/d in wk 6. This replacement linearly decreased milk yield (32.7 vs. 35.9 kg/d) and milk fat percentage and yield (2.32 vs. 3.22%, and 0.77 vs. 1.14 kg/d, respectively), but increased milk protein percentage and yield (3.80 vs. 3.04%, and 1.23 vs. 1.07 kg/d, respectively). This gradual replacement also linearly increased the daily averages of total volatile fatty acids (90 to 121.9 mM), acetate (53.9 to 66.8 mM), propionate (21.5 to 39.6 mM), and osmolality (277.7 to 293.8 mmol/kg) in the rumen and decreased the acetate to propionate ratio from 2.62 to 1.73. Replacing alfalfa hay with alfalfa pellets linearly increased blood lactate from 1.00 mM in wk 1 to a peak of 3.46 mM in wk 5. Induction of SARA in this study increased free rumen LPS concentration from 42,122 endotoxin unit (EU

  15. Residential indoor and outdoor coarse particles and associated endotoxin exposures

    Science.gov (United States)

    Wheeler, Amanda J.; Dobbin, Nina A.; Lyrette, Ninon; Wallace, Lance; Foto, Mark; Mallick, Ranjeeta; Kearney, Jill; Van Ryswyk, Keith; Gilbert, Nicolas L.; Harrison, Ian; Rispler, Kathleen; Héroux, Marie-Eve

    2011-12-01

    There is a growing body of evidence demonstrating that coarse particles (PM 10-2.5) have detrimental impacts upon health, especially for respiratory effects. There are limited data available for indoor residential exposures. Some data exist regarding the composition of this PM size fraction with emphasis on crustal elements and biological components. This study includes data from 146 homes sampled in Regina, Saskatchewan (SK) where 5-day integrated concurrent monitoring of indoor and outdoor coarse particles was conducted during the winter and summer of 2007. The coarse particle filters were subsequently analysed for endotoxin content to determine the contribution of this compound. Winter indoor geometric mean concentrations of coarse particles exceeded outdoor concentrations (3.73 μg m -3 vs 2.49 μg m -3; paired t-test p < 0.0001); however the reverse was found in summer (4.34 μg m -3 vs 8.82 μg m -3; paired t-test p < 0.0001). Linear regression indicated that winter predictors of indoor coarse particles were outdoor coarse particles, ventilation and presence of at least two or more occupants. During the summer, increased use of central air conditioning was associated with reduced coarse particles, while smoking and the presence of two or more occupants resulted in increased coarse particles. Endotoxin concentrations (EU μg -1) were lower indoors than outdoors in both seasons. Spatial variability of ambient coarse particles was assessed to determine the suitability of using a single monitoring station within a city to estimate exposure. The coefficients of variation between homes sampled simultaneously and the central monitoring station were calculated (median COV in summer = 15% and winter = 24%) and showed significant variability by week, especially during the summer months, suggesting a single site may be insufficient for characterizing exposure. Future studies should consider daily measurements per home to understand shorter term exposures and day to day

  16. Monocytes from cystic fibrosis patients are locked in an LPS tolerance state: down-regulation of TREM-1 as putative underlying mechanism.

    Directory of Open Access Journals (Sweden)

    Carlos del Fresno

    Full Text Available Cystic Fibrosis (CF is an inherited pleiotropic disease that results from abnormalities in the gene that codes for the chloride channel, Cystic Fibrosis Transmembrane Conductance Regulator (CFTR. CF patients are frequently colonized by several pathogens, but the mechanisms that allow colonization in spite of apparently functional immune systems are incompletely understood. In this paper we show that blood peripheral monocytes isolated from CF patients are found in an endotoxin tolerance state, yet this is not due to a deficient TLR activation. On the other hand, levels of the amplifier of inflammatory responses, TREM-1 (Triggering Receptor Expressed on Myeloid cells, are notably down-regulated in monocytes from patients, in comparison to those extracted from healthy volunteers. Furthermore, the soluble form of TREM-1 (sTREM-1 was not detected in the sera of patients. Additionally, and in strict contrast to patients who suffer from Chronic Obstructive Pulmonary Disease (COPD, CF monocytes challenged ex vivo with LPS neither up-regulated membrane-anchored TREM-1 nor sTREM-1. Finally, similar levels of PGE(2 expression and p65 translocation into the nucleus were found in both patients and healthy volunteers, thus suggesting that TREM-1 regulation is neither controlled by PGE(2 levels nor by p65 activation in this case. However, PU.1 translocation into the nucleus was significantly higher in CF monocytes than in controls, suggesting a role for this transcription factor in the control of TREM-1 expression. We conclude that down-regulation of TREM-1 expression in cystic fibrosis patients is at least partly responsible for the endotoxin tolerance state in which their monocytes are locked.

  17. LPS auto-calibration algorithm with predetermination of optimal zones.

    Science.gov (United States)

    Ruiz, Francisco Daniel; Ureña, Jesús; García, Juan C; Jiménez, Ana; Hernández, Alvaro; García, Juan J

    2011-01-01

    Accurate coordinates for active beacons placed in the environment are required in local positioning systems (LPS). These coordinates and the distances (or differences of distances) measured between the beacons and the mobile node to be localized are inputs to most trilateration algorithms. As a first approximation, such coordinates are obtained by means of manual measurements (a time-consuming and non-flexible method), or by using a calibration algorithm (i.e., automatic determination of beacon coordinates from ad hoc measurements). This paper presents a method to calibrate the beacons' positions in a LPS using a mobile receiver. The method has been developed for both, spherical and hyperbolic trilateration. The location of only three test points must be known a priori, while the position of the other test points can be unknown. Furthermore, the paper describes a procedure to estimate the optimal positions, or approximate areas in the coverage zone, where the test-points necessary to calibrate the ultrasonic LPS should be placed. Simulation and experimental results show the improvement achieved when these optimal test-points are used instead of randomly selected ones.

  18. LPS Auto-Calibration Algorithm with Predetermination of Optimal Zones

    Directory of Open Access Journals (Sweden)

    Juan C. García

    2011-10-01

    Full Text Available Accurate coordinates for active beacons placed in the environment are required in Local Positioning Systems (LPS. These coordinates and the distances (or differences of distances measured between the beacons and the mobile node to be localized are inputs to most trilateration algorithms. As a first approximation, such coordinates are obtained by means of manual measurements (a time-consuming and non-flexible method, or by using a calibration algorithm (i.e., automatic determination of beacon coordinates from ad hoc measurements. This paper presents a method to calibrate the beacons’ positions in a LPS using a mobile receiver. The method has been developed for both, spherical and hyperbolic trilateration. The location of only three test points must be known a priori, while the position of the other test points can be unknown. Furthermore, the paper describes a procedure to estimate the optimal positions, or approximate areas in the coverage zone, where the test-points necessary to calibrate the ultrasonic LPS should be placed. Simulation and experimental results show the improvement achieved when these optimal test-points are used instead of randomly selected ones.

  19. Endotoxin concentration in poultry houses for laying hens kept in cages or in alternative housing systems

    OpenAIRE

    2011-01-01

    Abstract 1. Endotoxins as components of organic dust may have adverse effects on the respiratory health of workers in poultry buildings. The move towards more welfare-friendly housing systems for layers may increase worker exposure to air contaminants due to the use of litter. 2. The endotoxin concentrations in the inhalable fraction of airborne dust (below 100 ?m) from cage and alternative system henhouses (on-floor, free range and aviaries) were compared under both experiment...

  20. Expression of Bacillus thuringiensis delta-endotoxin genes during vegetative growth.

    OpenAIRE

    Mettus, A M; Macaluso, A

    1990-01-01

    Bacillus thuringiensis delta-endotoxin (crystal protein) genes are normally expressed only during sporulation. It is possible to produce crystal protein during vegetative growth by placing B. thuringiensis crystal protein genes downstream of a strong vegetative promoter. By removing a possible transcriptional terminator of the tetracycline resistance gene of pBC16 and inserting a multiple cloning site, delta-endotoxin genes can be cloned downstream from the tetracycline resistance gene promot...

  1. The release of prostanoids during the acute pulmonary response to E. coli endotoxin in anaesthetized cats.

    OpenAIRE

    Coker, S. J.; Hughes, B.; Parratt, J. R.; Rodger, I. W.; Zeitlin, I. J.

    1983-01-01

    1 The administration of E. coli endotoxin (2 mg/kg i.v.) to anaesthetized cats results in a characteristic acute pulmonary response. This consists of increases in pulmonary artery pressure and airways resistance and a reduction in lung compliance. 2 Plasma concentrations of prostaglandin E2 (PGE2), PGF2 alpha, thromboxane B2 and 6-keto PGF1 alpha were measured by radioimmunoassay in aortic and pulmonary arterial blood samples before, during and after the acute pulmonary response to endotoxin....

  2. A non-chromatographic method for the removal of endotoxins from bacteriophages.

    Science.gov (United States)

    Branston, Steven D; Wright, Jason; Keshavarz-Moore, Eli

    2015-08-01

    The Ff filamentous bacteriophages show potential as a new class of therapeutics, displaying utility in materials science as well as pharmaceutical applications. These phages are produced by the infection of E. coli, a Gram-negative bacterium which unavoidably sheds endotoxins into the extracellular space during growth. Since endotoxin molecules are highly immunoreactive, separation from the phage product is of critical importance, particularly those developed for human therapeutic use. The properties of M13, one of the Ff group, present a purification challenge chiefly because the standard scalable method for endotoxin removal from proteins-anion exchange chromatography-is not applicable due to pI similarity between the particles. This article examines the potential of polyethylene glycol (PEG)-NaCl precipitation as a scalable method for the separation of endotoxins from phage M13. Precipitation of M13 by 2% (w/v) PEG 6 000, 500 mM NaCl reduced endotoxin contamination of the phage product by 88%, but additional precipitation rounds did not maintain this proportional decrease. Dynamic light scattering was subsequently used to determine the effectiveness of a detergent to disassociate endotoxin molecules from M13. As a result, PEG-NaCl precipitation was supplemented with up to 2% (v/v) Triton X-100 to improve separation. A 5.7 log10 reduction in endotoxin concentration was achieved over three rounds of precipitation whilst retaining over 97% of the phage. This method compares favorably with the well-known ATPS (Triton X-114) technique for endotoxin removal from protein solutions. © 2015 Wiley Periodicals, Inc.

  3. Case Report. The Use of Endotoxin Adsorption in Extracorporeal Blood Purification Techniques. A Case Report

    Directory of Open Access Journals (Sweden)

    Hadade Adina N.

    2017-04-01

    Full Text Available Sepsis and septic shock are major healthcare problems, resulting in high morbidity and mortality. The Surviving Sepsis Campaign (SSC, which standardised the approach to sepsis, was recently updated. Strategies to decrease the systemic inflammatory response have been proposed to modulate organ dysfunctions. Endotoxin, derived from the membrane of Gram-negative bacteria, is considered a major factor in the pathogenesis of sepsis. Endotoxin adsorption, if effective, has the potential to reduce the biological cascade of Gram-negative sepsis.

  4. Removal of endotoxin from deionized water using micromachined silicon nanopore membranes

    Science.gov (United States)

    Smith, Ross A.; Goldman, Ken; Fissell, William H.; Fleischman, Aaron J.; Zorman, Christian A.; Roy, Shuvo

    2011-05-01

    Endotoxins are lipopolysaccharide components of the cell membrane of Gram-negative bacteria that trigger the body's innate immune system and can cause shock and death. Water for medical therapy, including parenteral and dialysate solutions, must be free of endotoxin. This purity is challenging to achieve as many Gram-negative bacteria are endemic in the environment, and can thrive in harsh, nutrient-poor conditions. Current methods for removing endotoxin include distillation and reverse osmosis, both of which are resource intensive processes. Membranes that present an absolute barrier to macromolecular passage may be capable of delivering pure water for biomedical applications. In this work, endotoxin has been filtered from aqueous solutions using silicon nanopore membranes (SNMs) with monodisperse pore size distributions. SNMs with critical pore sizes between 26 and 49 nm were challenged with solutions of deionized water spiked with endotoxin and with Pseudomonas cepacia. The filtrate produced by the SNM from Pseudomonas-contaminated water had <1.0 endotoxin unit (EU) ml-1, which meets standards for dialysate purity. This approach suggests a technique for single-step cleanup of heavily contaminated water that may be suitable for field or clinical use.

  5. Modeling human health risks of airborne endotoxin in homes during the winter and summer seasons

    Energy Technology Data Exchange (ETDEWEB)

    Liao, Vivian Hsiu-Chuan, E-mail: vivianliao@ntu.edu.tw [Department of Bioenvironmental Systems Engineering, National Taiwan University, Taipei 106, Taiwan (China); Chio, Chia-Pin; Chou, Wei-Chun; Ju, Yun-Ru; Liao, Chung-Min [Department of Bioenvironmental Systems Engineering, National Taiwan University, Taipei 106, Taiwan (China)

    2010-03-01

    Endotoxin, a component of gram-negative bacterial cell walls, is a pro-inflammatory agent that induces local and systemic inflammatory responses in normal subjects which can contribute to the risk of developing asthma and chronic obstructive lung diseases. A probabilistic approach linking models of exposure, internal dosimetry, and health effects was carried out to quantitatively assess the potential inhalation risk of airborne endotoxin in homes during the winter and summer seasons. Combining empirical data and modeling results, we show that the half-maximum effect of the endotoxin dose (ED50) was estimated to be 707.9 (95% confidence interval (CI): 308.8-1287.0) endotoxin units (EU) for body temperature change, 481.8 (95% CI: 333.2-630.3) EU for elevation of neutrophils, and 1174.5 (95% CI: 816.0-1532.9) EU for elevation of the cytokine, interleukin-6. Our study also suggests that airborne endotoxin in homes may pose potential risks, and a higher risk for elevation of neutrophils and cytokine interleukin-6 appeared in winter season than in summer. Our study offers a risk-management framework for discussion of future studies of human respiratory exposure to airborne endotoxin.

  6. Innate immune dysfunction in the neonatal rat following prenatal endotoxin exposure.

    Science.gov (United States)

    Hodyl, Nicolette A; Krivanek, Klara M; Clifton, Vicki L; Hodgson, Deborah M

    2008-11-15

    The efficacy of the neonatal innate immune system to respond to bacterial exposure following maternal infection is of great interest, as the neonatal period is one of relative immune immaturity, and is associated with high rates of morbidity and mortality. This study aimed to determine the response to an in-vivo endotoxin challenge in the neonatal period following prenatal exposure to bacterial endotoxin. Pregnant Fischer 344 dams received either endotoxin or the vehicle on gestational days 16, 18 and 20 (term=23 days). The neonatal (5 day) offspring were then exposed to an endotoxin challenge; blood was collected at baseline or at 4 h for analysis of blood cell counts, corticosterone, TNF alpha and IL-1 beta, levels. The neonatal rat pups responded to the challenge with significantly reduced corticosterone, TNF alpha and IL-1 beta levels compared to controls (p<0.003). Monocyte, neutrophil and eosinophil counts were also significantly reduced in the prenatal endotoxin offspring compared to controls (p<0.02). While the immune system is functionally immature in the neonatal period, these results suggest that prenatal infection may further reduce the capacity of the innate neonatal immune system to respond to endotoxin, leaving offspring more vulnerable to pathogenic invasion in neonatal life.

  7. Comparison of Different Irrigants in the Removal of Endotoxins and Cultivable Microorganisms from Infected Root Canals.

    Science.gov (United States)

    Valera, Marcia Carneiro; Cardoso, Flávia Goulart da Rosa; Chung, Adriana; Xavier, Ana Cláudia Carvalho; Figueiredo, Mariana Diehl; Martinho, Frederico Canato; Palo, Renato Miotto

    2015-01-01

    This study was conducted to compare the effectiveness of different irrigants used to remove endotoxins and cultivable microorganisms during endodontic therapy. Forty root canals were contaminated and divided into groups according to the irrigant: 2% NaOCl + surfactant, 2% CHX, 2.5% NaOCl, and pyrogen-free saline solution (control). Samples were collected after root canal contamination (S1), after instrumentation (S2), and 7 days after instrumentation (S3). Microorganisms and endotoxins were recovered from 100% of the contaminated root canals (S1). At S2, 2% NaOCl + surfactant, 2% CHX, and 2.5% NaOCl were able to completely eliminate cultivable microorganisms. At S3, both 2% CHX and 2.5% NaOCl were effective in preventing C. albicans and E. coli regrowth, but E. faecalis was still detected. No microorganism species was recovered from root canals instrumented with 2% NaOCl + surfactant. At S2, a higher percentage value of endotoxin reduction was found for 2% NaOCl + surfactant (99.3%) compared to 2% CHX (98.9%) and 2.5% NaOCl (97.18%) (p endotoxins. All irrigants tested were effective in reducing microorganisms and endotoxins from root canals. Moreover, 2% NaOCl + surfactant was the most effective irrigant against endotoxins and regrowth of microorganisms.

  8. Structural changes in the rat middle ear mucosa due to endotoxin and eustachian tube obstruction.

    Science.gov (United States)

    Nell, M J; Grote, J J

    1999-01-01

    The middle ears of 48 rats were used to examine the effects of endotoxin injection, eustachian tube obstruction or a combination of eustachian tube obstruction and endotoxin injection. Animals were killed after 1, 2, 4, or 12 weeks and the middle ears processed for light and scanning electron microscopy. Compared to the normal middle ear mucosa, the epithelial layer was more pseudostratified, cuboidal or cylindrical after endotoxin injection or obstruction of the eustachian tube. In the early phase, numerous ciliated cells occurred in areas originally almost devoid of these cells. At 3 months, degeneration of ciliated cells was observed. The combination of eustachian tube obstruction and endotoxin injection also induced a more pseudostratified, cuboidal or cylindrical epithelium with an increased number of goblet cells. However, an early decrease occurred in the number of ciliated cells in the tympanic orifice of the eustachian tube. Furthermore, inflammatory cells, mainly PMNs, macrophages and lymphocytes, invaded the subepithelial layer after eustachian tube obstruction and endotoxin injection. These structural changes resulted in an impairment of the mucociliary transport system for clearance of the middle ear cavity. For this reason we believe that both endotoxin and eustachian tube obstruction or dysfunction play an important role in inducing persistent mucosal changes in the middle ear cavity, thereby prolonging otitis media with effusion.

  9. Experimental Study on Inactivation of Bacterial Endotoxin by Using Dielectric Barrier Discharge

    Science.gov (United States)

    Shi, Xingmin; Li, Yaxi; Zhang, Guanjun; Ma, Yue; Shao, Xianjun

    2011-12-01

    The low-temperature plasma (LTP) generated by dielectric barrier discharge (DBD) was used to sterilize the E.coli endotoxin, which is usually difficult to kill by traditional methods. Three different concentrations of bacterial endotoxin (1 EU/mL, 0.5 EU/mL and 0.25 EU/mL) were treated by LTP for different time (20 s, 40 s and 60 s). Tachypleus amebocyte lysate (TAL) method was employed to detect the concentration variation of bacterial endotoxin before and after the plasma treatment, and endotoxic shock mice model was used to evaluate the inactivation effects of LTP on endotoxin for further study. Experimental results demonstrated that, DBD plasma can inactivate the bacterial endotoxin quickly and effectively, and when the LTP treatment time was increased, the concentrations of bacterial endotoxin decreased gradually (after 60 s plasma treatment, its inactivation effect was beyond the Chinese pharmacopoeia standard), and the average survival time of mice gradually extended. The possible inactivation mechanisms are proposed to be related to reactive oxygen species (ROSs).

  10. Procalcitonin neutralizes bacterial LPS and reduces LPS-induced cytokine release in human peripheral blood mononuclear cells.

    Science.gov (United States)

    Matera, Giovanni; Quirino, Angela; Giancotti, Aida; Pulicari, Maria Concetta; Rametti, Linda; Rodríguez, Maria Luz; Liberto, Maria Carla; Focà, Alfredo

    2012-05-08

    Procalcitonin (PCT) is a polypeptide with several cationic aminoacids in its chemical structure and it is a well known marker of sepsis. It is now emerging that PCT might exhibit some anti-inflammatory effects. The present study, based on the evaluation of the in vitro interaction between PCT and bacterial lipopolisaccharide (LPS), reports new data supporting the interesting and potentially useful anti-inflammatory activity of PCT. PCT significantly decreased (p cytokine release were studied in human peripheral blood mononuclear cells (PBMC). When LPS was pre-incubated for 30 minutes with different concentrations of PCT, the release of interleukin-10 (IL-10) and tumor necrosis factor alpha (TNFα) by PBMC decreased in a concentration-dependent manner after 24 hours for IL-10 and 4 hours for TNFα. The release of monocyte chemotactic protein-1 (MCP-1) exhibited a drastic reduction at 4 hours for all the PCT concentrations assessed, whereas such decrease was concentration-dependent after 24 hours. This study provides the first evidence of the capability of PCT to directly neutralize bacterial LPS, thus leading to a reduction of its major inflammatory mediators.

  11. Enhanced innate immune responsiveness and intolerance to intestinal endotoxins in human biliary epithelial cells contributes to chronic cholangitis.

    Science.gov (United States)

    Mueller, Tobias; Beutler, Claudia; Picó, Almudena Hurtado; Shibolet, Oren; Pratt, Daniel S; Pascher, Andreas; Neuhaus, Peter; Wiedenmann, Bertram; Berg, Thomas; Podolsky, Daniel K

    2011-11-01

    Pattern recognition receptors (PRRs) orchestrate the innate immune defence in human biliary epithelial cells (BECs). Tight control of PRR signalling provides tolerance to physiological amounts of intestinal endotoxins in human bile to avoid constant innate immune activation in BECs. We wanted to determine whether inappropriate innate immune responses to intestinal endotoxins contribute to the development and perpetuation of chronic biliary inflammation. We examined PRR-mediated innate immune responses and protective endotoxin tolerance in primary BECs isolated from patients with primary sclerosing cholangitis (PSC), alcoholic liver disease and patients without chronic liver disease. Expression studies comprised northern blots, RT-PCR, Western blots and immunocytochemistry. Functional studies comprised immuno-precipitation Western blots, FACS for endotoxin uptake, and NF-κB activation assays and ELISA for secreted IL-8 and tumour necrosis factor (TNF)-α. Primary BECs from explanted PSC livers showed reversibly increased TLR and NOD protein expression and activation of the MyD88/IRAK signalling complex. Consecutively, PSC BECs exhibited inappropriate innate immune responses to endotoxins and did not develop immune tolerance after repeated endotoxin exposures. This endotoxin hyper-responsiveness was probably because of the stimulatory effect of abundantly expressed IFN-γ and TNF-α in PSC livers, which stimulated TLR4-mediated endotoxin signalling in BECs, leading to increased TLR4-mediated endotoxin incorporation and impaired inactivation of the TLR4 signalling cascade. As TNF-α inhibition partly restored protective innate immune tolerance, endogenous TNF-α secretion probably contributed to inappropriate endotoxin responses in BECs. Inappropriate innate immune responses to intestinal endotoxins and subsequent endotoxin intolerance because of enhanced PRR signalling in BECs probably contribute to chronic cholangitis. © 2011 John Wiley & Sons A/S.

  12. Escin Increases the Survival Rate of LPS-Induced Septic Mice Through Inhibition of HMGB1 Release from Macrophages

    Directory of Open Access Journals (Sweden)

    Yajun Cheng

    2015-07-01

    Full Text Available Background: Previous studies have described the effects of Escin on improving the survival rate of endotoxemic animals. The purpose of this study was to explore the molecular mechanisms of this potentially beneficial treatment. Methods: First, the survival rate of endotoxemic mice was monitored for up to 2 weeks after Escin pretreatment, Escin post-treatment, or Escin post-treatment + rHMGB1. The effects of Escin on the release of pro-inflammatory cytokines such as TNF-a, IL-1ß, IL-6 and HMGB1 in the serum of endotoxemic mice and LPS-induced macrophages were evaluated by ELISA. Furthermore, the mRNA and protein levels of HMGB1 in LPS-induced macrophages were measured by qRT-PCR and Western blot, respectively. Additionally, the release of pro-inflammatory cytokines such as TNF-a, IL-1ß, IL-6 was evaluated by ELISA in rHMGB1-induced macrophages. Finally, the protein levels and the activity of NF-κB in macrophages were checked by Western blot and ELISA, respectively. Results: Both pretreatment and post-treatment with Escin could improve the survival rate of endotoxemic mice, while exogenous rHMGB1 reversed this effect. In addition, Escin decreased the level of the pro-inflammatory cytokines TNF-a, IL-1ß, IL-6 and HMGB1 in endotoxemic mice and in LPS-induced macrophages. Escin could also inhibit the mRNA levels and activity of HMGB1. The release of the pro-inflammatory cytokines TNF-a, IL-1ß, IL-6 could be suppressed in rHMGB1-induced macrophages by Escin. Finally, Escin could suppress the activation of NF-κB in LPS-induced macrophages. Conclusion: Escin could improve the survival of mice with LPS-induced endotoxemia. This effect maybe meditated by reducing the release of HMGB1, resulting in the suppression of the release of pro-inflammatory cytokines.

  13. Leakage of Microbial Endotoxin through the Implant-Abutment Interface in Oral Implants: An In Vitro Study

    Directory of Open Access Journals (Sweden)

    Rhoodie Garrana

    2016-01-01

    Full Text Available Background. Endotoxin initiates osteoclastic activity resulting in bone loss. Endotoxin leakage through implant abutment connections negatively influences peri-implant bone levels. Objectives. (i To determine if endotoxin can traverse different implant-abutment connection (IAC designs; (ii to quantify the amount of endotoxins traversing the IAC; (iii to compare the in vitro comportments of different IACs. Materials and Methods. Twenty-seven IACs were inoculated with E. coli endotoxin. Six of the twenty-seven IACs were external connections from one system (Southern Implants and the remaining twenty-one IACs were made up of seven internal IAC types from four different implant companies (Straumann, Ankylos, and Neodent, Southern Implants. Results. Of the 27 IACs tested, all 6 external IACs leaked measurable amounts of endotoxin. Of the remaining 21 internal IACs, 9 IACs did not show measurable leakage whilst the remaining 12 IACs leaked varying amounts. The mean log endotoxin level was significantly higher for the external compared to internal types (p=0.015. Conclusion. Within the parameters of this study, we can conclude that endotoxin leakage is dependent on the design of the IAC. Straumann Synocta, Straumann Cross-fit, and Ankylos displayed the best performances of all IACs tested with undetectable leakage after 7 days. Each of these IACs incorporated a morse-like component in their design. Speculation still exists over the impact of IAC endotoxin leakage on peri-implant tissues in vivo; hence, further investigations are required to further explore this.

  14. Leakage of Microbial Endotoxin through the Implant-Abutment Interface in Oral Implants: An In Vitro Study.

    Science.gov (United States)

    Garrana, Rhoodie; Mohangi, Govindrau; Malo, Paulo; Nobre, Miguel

    2016-01-01

    Background . Endotoxin initiates osteoclastic activity resulting in bone loss. Endotoxin leakage through implant abutment connections negatively influences peri-implant bone levels. Objectives . (i) To determine if endotoxin can traverse different implant-abutment connection (IAC) designs; (ii) to quantify the amount of endotoxins traversing the IAC; (iii) to compare the in vitro comportments of different IACs. Materials and Methods . Twenty-seven IACs were inoculated with E. coli endotoxin. Six of the twenty-seven IACs were external connections from one system (Southern Implants) and the remaining twenty-one IACs were made up of seven internal IAC types from four different implant companies (Straumann, Ankylos, and Neodent, Southern Implants). Results . Of the 27 IACs tested, all 6 external IACs leaked measurable amounts of endotoxin. Of the remaining 21 internal IACs, 9 IACs did not show measurable leakage whilst the remaining 12 IACs leaked varying amounts. The mean log endotoxin level was significantly higher for the external compared to internal types ( p = 0.015). Conclusion . Within the parameters of this study, we can conclude that endotoxin leakage is dependent on the design of the IAC. Straumann Synocta, Straumann Cross-fit, and Ankylos displayed the best performances of all IACs tested with undetectable leakage after 7 days. Each of these IACs incorporated a morse-like component in their design. Speculation still exists over the impact of IAC endotoxin leakage on peri-implant tissues in vivo; hence, further investigations are required to further explore this.

  15. Dose response of endotoxin on hepatocyte and muscle mitochondrial respiration in vitro.

    Science.gov (United States)

    Jeger, Victor; Brandt, Sebastian; Porta, Francesca; Jakob, Stephan M; Takala, Jukka; Djafarzadeh, Siamak

    2015-01-01

    Results on mitochondrial dysfunction in sepsis are controversial. We aimed to assess effects of LPS at wide dose and time ranges on hepatocytes and isolated skeletal muscle mitochondria. Human hepatocellular carcinoma cells (HepG2) were exposed to placebo or LPS (0.1, 1, and 10 μg/mL) for 4, 8, 16, and 24 hours and primary human hepatocytes to 1 μg/mL LPS or placebo (4, 8, and 16 hours). Mitochondria from porcine skeletal muscle samples were exposed to increasing doses of LPS (0.1-100 μg/mg) for 2 and 4 hours. Respiration rates of intact and permeabilized cells and isolated mitochondria were measured by high-resolution respirometry. In HepG2 cells, LPS reduced mitochondrial membrane potential and cellular ATP content but did not modify basal respiration. Stimulated complex II respiration was reduced time-dependently using 1 μg/mL LPS. In primary human hepatocytes, stimulated mitochondrial complex II respiration was reduced time-dependently using 1 μg/mL LPS. In isolated porcine skeletal muscle mitochondria, stimulated respiration decreased at high doses (50 and 100 μg/mL LPS). LPS reduced cellular ATP content of HepG2 cells, most likely as a result of the induced decrease in membrane potential. LPS decreased cellular and isolated mitochondrial respiration in a time-dependent, dose-dependent and complex-dependent manner.

  16. Effects of immune challenge on concentrations of serum insulin-like growth factor-I and growth performance in pigs.

    OpenAIRE

    Hevener, W; Routh, P A; Almond, G W

    1999-01-01

    This study was designed to determine the long-term effects of repeated endotoxin treatment or immunization against human serum albumin on concentrations of serum insulin-like growth factor-I (IGF-I) and other indicators of growth performance in growing pigs. Thirty gilts (38.5 +/- 0.9 kg) were randomly assigned to 5 treatment groups (n = 6 animals/group): 1) lipopolysaccharide injections, 2) lipopolysaccharide pair-fed, 3) human serum albumin immunization, 4) human serum albumin pair-fed, and...

  17. Endotoxin-Induced Inflammation Suppresses the Effect of Melatonin on the Release of LH from the Ovine Pars Tuberalis Explants—Ex Vivo Study

    Directory of Open Access Journals (Sweden)

    Karolina Wojtulewicz

    2017-11-01

    Full Text Available The secretion of the hormone melatonin reliably reflects environmental light conditions. Among numerous actions, in seasonal breeders, melatonin may regulate the secretion of the gonadotropins acting via its corresponding receptors occurring in the Pars Tuberalis (PT. However, it was previously found that the secretory activity of the pituitary may be dependent on the immune status of the animal. Therefore, this study was designed to determine the role of melatonin in the modulation of luteinizing hormone (LH secretion from the PT explants collected from saline- and endotoxin-treated ewes in the follicular phase of the oestrous cycle. Twelve Blackhead ewes were sacrificed 3 h after injection with lipopolysaccharide (LPS; 400 ng/kg or saline, and the PTs were collected. Each PT was cut into 4 explants, which were then divided into 4 groups: I, incubated with ‘pure’ medium 199; II, treated with gonadotropin-releasing hormone (GnRH (100 pg/mL; III, treated with melatonin (10 nmol/mL; and IV, incubated with GnRH and melatonin. Melatonin reduced (p < 0.05 GnRH-induced secretion of LH only in the PT from saline-treated ewes. Explants collected from LPS-treated ewes were characterized by lower (p < 0.05 GnRH-dependent response in LH release. It was also found that inflammation reduced the gene expression of the GnRH receptor and the MT1 melatonin receptors in the PT. Therefore, it was shown that inflammation affects the melatonin action on LH secretion from the PT, which may be one of the mechanisms via which immune/inflammatory challenges disturb reproduction processes in animals.

  18. Effects in man and rabbits of inhalation of cotton dust or extracts and purified endotoxins1

    Science.gov (United States)

    Cavagna, G.; Foá, V.; Vigliani, E. C.

    1969-01-01

    Cavagna, G., Foá V., and Vigliani, E. C. (1969).Brit. J. industr. Med.,26, 314-321. Effects in man and rabbits of inhalation of cotton dust or extracts and purified endotoxins. The incidence of byssinosis in workers in a cotton card-room, where the airborne concentration of bacterial endotoxins was 7·2 μg./m.3, was 32%; and 47% in a hemp card-room where the endotoxin concentration was 8·7 μg./m.3; no cases were observed among workers exposed to only traces of endotoxins. The effect of the inhalation by aerosol of purified Escherichia coli endotoxin on F.E.V.1·0 and F.V.C. was studied in normal subjects and in patients with chronic bronchitis. A significant reduction in F.E.V.1·0 lasting more than 6 hours was observed in two out of eight normal subjects, in one out of three subjects with chronic bronchitis inhaling 80 μg. endotoxin, and in one out of four subjects with chronic bronchitis inhaling 40 μg. endotoxin. These results show that the inhalation of bacterial endotoxin can produce, in some individuals, changes in F.E.V.1·0 similar to those experienced on Mondays by some card-room workers. A study of the mechanism of pathogenesis of inhaled bacterial endotoxins was carried out on rabbits subjected for 20 weeks to aerosols of purified E. coli endotoxin (20 μg./day) and cotton extract (2 mg./day). This treatment produced patterns of bronchitis: i.e., a increase in the respiratory tract fluid with increased protein content and characteristic histopathological changes. The bronchitis occurred after the appearance of cross-reacting circulating antibodies against E. coli endotoxin and cotton extract. These antibodies were detected with the haemagglutination tests after the first three weeks of treatment, and in subsequent weeks reached progressively higher titres, up to a maximum of 1:512. A challenging aerosol of 0·1 mg. E. coli endotoxin in two rabbits and 10 mg. cotton extract in another two of the rabbits treated as above produced a marked increase in

  19. Autotaxin and Endotoxin-Induced Acute Lung Injury.

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    Marios-Angelos Mouratis

    Full Text Available Acute Lung Injury (ALI is a life-threatening, diffuse heterogeneous lung injury characterized by acute onset, pulmonary edema and respiratory failure. Lipopolysaccharide (LPS is a common cause of both direct and indirect lung injury and when administered to a mouse induces a lung phenotype exhibiting some of the clinical characteristics of human ALI. Here, we report that LPS inhalation in mice results in increased bronchoalveolar lavage fluid (BALF levels of Autotaxin (ATX, Enpp2, a lysophospholipase D largely responsible for the conversion of lysophosphatidylcholine (LPC to lysophosphatidic acid (LPA in biological fluids and chronically inflamed sites. In agreement, gradual increases were also detected in BALF LPA levels, following inflammation and pulmonary edema. However, genetic or pharmacologic targeting of ATX had minor effects in ALI severity, suggesting no major involvement of the ATX/LPA axis in acute inflammation. Moreover, systemic, chronic exposure to increased ATX/LPA levels was shown to predispose to and/or to promote acute inflammation and ALI unlike chronic inflammatory pathophysiological situations, further suggesting a differential involvement of the ATX/LPA axis in acute versus chronic pulmonary inflammation.

  20. Autotaxin and Endotoxin-Induced Acute Lung Injury

    Science.gov (United States)

    Oikonomou, Nikos; Katsifa, Aggeliki; Prestwich, Glenn D.; Kaffe, Eleanna; Aidinis, Vassilis

    2015-01-01

    Acute Lung Injury (ALI) is a life-threatening, diffuse heterogeneous lung injury characterized by acute onset, pulmonary edema and respiratory failure. Lipopolysaccharide (LPS) is a common cause of both direct and indirect lung injury and when administered to a mouse induces a lung phenotype exhibiting some of the clinical characteristics of human ALI. Here, we report that LPS inhalation in mice results in increased bronchoalveolar lavage fluid (BALF) levels of Autotaxin (ATX, Enpp2), a lysophospholipase D largely responsible for the conversion of lysophosphatidylcholine (LPC) to lysophosphatidic acid (LPA) in biological fluids and chronically inflamed sites. In agreement, gradual increases were also detected in BALF LPA levels, following inflammation and pulmonary edema. However, genetic or pharmacologic targeting of ATX had minor effects in ALI severity, suggesting no major involvement of the ATX/LPA axis in acute inflammation. Moreover, systemic, chronic exposure to increased ATX/LPA levels was shown to predispose to and/or to promote acute inflammation and ALI unlike chronic inflammatory pathophysiological situations, further suggesting a differential involvement of the ATX/LPA axis in acute versus chronic pulmonary inflammation. PMID:26196781

  1. Does the Reciproc file remove root canal bacteria and endotoxins as effectively as multifile rotary systems?

    Science.gov (United States)

    Marinho, A C S; Martinho, F C; Gonçalves, L M; Rabang, H R C; Gomes, B P F A

    2015-06-01

    To evaluate the effectiveness of Reciproc for the removal of cultivable bacteria and endotoxins from root canals in comparison with multifile rotary systems. The root canals of forty human single-rooted mandibular pre-molars were contaminated with an Escherichia coli suspension for 21 days and randomly assigned to four groups according to the instrumentation system: GI - Reciproc (VDW); GII - Mtwo (VDW); GIII - ProTaper Universal (Dentsply Maillefer); and GIV -FKG Race(™) (FKG Dentaire) (n = 10 per group). Bacterial and endotoxin samples were taken with a sterile/apyrogenic paper point before (s1) and after instrumentation (s2). Culture techniques determined the colony-forming units (CFU) and the Limulus Amebocyte Lysate assay was used for endotoxin quantification. Results were submitted to paired t-test and anova. At s1, bacteria and endotoxins were recovered in 100% of the root canals investigated (40/40). After instrumentation, all systems were associated with a highly significant reduction of the bacterial load and endotoxin levels, respectively: GI - Reciproc (99.34% and 91.69%); GII - Mtwo (99.86% and 83.11%); GIII - ProTaper (99.93% and 78.56%) and GIV - FKG Race(™) (99.99% and 82.52%) (P endotoxin removal (P > 0.01). The reciprocating single file, Reciproc, was as effective as the multifile rotary systems for the removal of bacteria and endotoxins from root canals. © 2014 International Endodontic Journal. Published by John Wiley & Sons Ltd.

  2. Cardiorenal Syndrome Type 5 in Sepsis: Role of Endotoxin in Cell Death Pathways and Inflammation

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    Grazia Maria Virzì

    2016-12-01

    Full Text Available Background/Aims: Cardiorenal Syndrome Type 5 (CRS Type 5 is characterized by concomitant cardiac and renal dysfunction in the setting of different systemic disorders, such as sepsis. In this study, we investigated the possible relationship between endotoxin levels, renal cell death and inflammation in septic patients with CRS Type 5. Methods: We enrolled 11 patients with CRS Type 5. CRS Type 5 was defined according to the current classification system. AKI was defined by Acute Kidney Injury Network (AKIN criteria. Acute cardiac dysfunction was documented by echocardiography as acute left and/or right ventricular dysfunction leading to decreased ejection fraction. Endotoxin activity was measured by the Endotoxin Activity Assay (EAA. Plasma from CRS Type 5 patients was incubated with renal tubular cells (RTCs and cell death levels were evaluated. Plasma cytokines levels were measured as well. Results: Accordingly to EAA levels, patients were divided into two groups: 45.4% of patients had low endotoxin activity level (negative EAA, while 54.5% of patients showed high endotoxin activity (positive EAA. RTCs incubated with plasma from EAA positive patients showed significantly higher apoptosis levels and higher caspase-3 activation compared to cells incubated with plasma from EAA negative patients, and a significant positive correlation was observed between EAA levels and RTC apoptosis levels. Furthermore, IL-6 and IFN-γ levels were significantly higher in CRS Type 5 patients with positive EAA. Conclusion: Our data suggest a possible relationship between endotoxin levels and renal cell death in septic patients with CRS Type 5. Furthermore, this study highlights the presence of renal apoptosis, the immune deregulation and the strong inflammation in CRS Type 5 patients, especially in those with high endotoxin activity.

  3. Dialysate bacterial endotoxin as a prognostic indicator of peritoneal dialysis related peritonitis.

    Science.gov (United States)

    Szeto, Cheuk-Chun; Lai, Ka-Bik; Chow, Kai-Ming; Kwan, Bonnie Ching-Ha; Law, Man-Ching; Pang, Wing-Fai; Ma, Terry King-Wing; Leung, Chi-Bon; Li, Philip Kam-Tao

    2016-12-01

    Peritonitis is the major complication of peritoneal dialysis (PD). The aim of our