WorldWideScience

Sample records for bacterial endotoxin challenge

  1. 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.

  2. DMPD: Targeting bacterial endotoxin: two sides of a coin. [Dynamic Macrophage Pathway CSML Database

    Lifescience Database Archive (English)

    Full Text Available 17405910 Targeting bacterial endotoxin: two sides of a coin. Bosshart H, Heinzelman...n M. Ann N Y Acad Sci. 2007 Jan;1096:1-17. (.png) (.svg) (.html) (.csml) Show Targeting bacterial endotoxin:... two sides of a coin. PubmedID 17405910 Title Targeting bacterial endotoxin: two sides of a coin. Authors Bo

  3. Cotton bacterial endotoxin assessed by electron microscopy.

    OpenAIRE

    Helander, I; Lounatmaa, K.

    1981-01-01

    A piece of bale cotton was incubated in nutrient broth. Electron microscopic inspection of the cotton and the broth showed Gram-negative bacteria with long flagella, loosely attached to the cotton fibres. Large amounts of endotoxin liberating from these bacteria were visible in the growth medium.

  4. 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.

  5. Impact of endotoxin challenge in obese pigs.

    Science.gov (United States)

    Duburcq, Thibault; Hubert, Thomas; Saint-Léger, Pierre; Mangalaboyi, Jacques; Favory, Raphael; Gmyr, Valery; Quintane, Laurence; Tailleux, Anne; Staels, Bart; Tournoys, Antoine; Pattou, François; Jourdain, Mercé

    2014-06-01

    Studies exploring the influence of obesity on septic shock remain limited and controversial. Pigs were chosen as a clinically relevant species, resembling to humans in various functions. We hypothesize obesity may impair porcine acute endotoxic shock. Four groups of five "Yucatan" minipigs were studied: lean and obese control groups, lean lipopolysaccharide (LPS) group receiving Escherichia coli endotoxin (LPS) and obese LPS group receiving the same endotoxin dose. We measured hemodynamic and oxygenation parameters, skin microvascular blood flow at rest and during reactive hyperemia, von Willebrand factor, tumor necrosis factor α, and interleukin 6. All measurements were performed at baseline and at 30, 60, 90, 150, and 300 min. Results were given as median with 25th to 75th interquartile range. Control groups remained stable during the study period. In LPS groups, administration of endotoxin resulted in a typical hypokinetic shock. In obese LPS group at 300 min, we observed a significant impairment of cardiac index (1.2 [1.06-1.45] vs. 1.7 [1.57-1.97] L/min per m, P = 0.008) compared with the lean LPS group; moreover, pulmonary hypertension (mean arterial pressure: 42 [39-47] vs. 32 [28-34] mmHg, P = 0.008), hypoxemia (partial pressure of oxygen: 216 [178-262] vs. 325 [285-414] mmHg, P = 0.02), and lactate levels (5.8 [4.2-6.8] vs. 3.9 [2.2-5.5] mmol/L, P = 0.04) were significantly higher compared with the lean LPS group. Throughout the study, rest flow and peak flow during reactive hyperemia were more decreased in the obese LPS group. Compared with the lean LPS group, tumor necrosis factor α levels at 60 min (269 [178-428] vs. 126 [105-166] ng/mL, P = 0.03) and interleukin 6 levels at 300 min (101 [61-142] vs. 52 [36-64] ng/mL, P = 0.03) were significantly higher in the obese LPS group. In our model of endotoxic shock, obese pigs developed a more severe hemodynamic failure with pronounced microcirculatory dysfunction and proinflammatory response. PMID:24569508

  6. Experimental Study on Inactivation of Bacterial Endotoxin by Using Dielectric Barrier Discharge%Experimental Study on Inactivation of Bacterial Endotoxin by Using Dielectric Barrier Discharge

    Institute of Scientific and Technical Information of China (English)

    石兴民; 李娅西; 张冠军; 马跃; 邵先军

    2011-01-01

    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 af- ter 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).

  7. Concentration, physical state, and purity of bacterial endotoxin affect its detoxification by ionizing radiation

    Energy Technology Data Exchange (ETDEWEB)

    Csako, G.; Tsai, C.M.; Hochstein, H.D.; Elin, R.J.

    1986-11-01

    Increasing concentrations of a highly purified bacterial lipopolysaccharide preparation, the U.S. Reference Standard Endotoxin, were exposed to increasing doses of ionizing radiation from a 60Co source. At identical radiation doses both the structural change and Limulus amebocyte lysate (LAL) reactivity were progressively smaller with increasing concentrations of the lipopolysaccharide in an aqueous medium. Under the experimental conditions used, there was a linear relationship between the endotoxin concentration and radiation dose for the structural changes. In contrast to endotoxin in aqueous medium, endotoxin irradiated in its dry state showed no decrease in LAL reactivity and rabbit pyrogenicity. Endotoxin exposed to radiation in water in the presence of albumin showed a much smaller decrease in LAL and pyrogenic activities than expected. The results show that the concentration, physical state, and purity of endotoxin influence its structural and functional alteration by ionizing radiation.

  8. Short time bacterial endotoxins test for positron emission tomography by means of positively charged filters

    International Nuclear Information System (INIS)

    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 [18F]FDG and [18F]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)

  9. 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.

  10. 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)

  11. Effect of cisapride on intestinal bacterial and endotoxin translocation in cirrhosis

    Institute of Scientific and Technical Information of China (English)

    Shun-Cai Zhang; Wei Wang; Wei-Ying Ren; Bo-Ming He; Kang Zhou; Wu-Nan Zhu

    2003-01-01

    AIM: To investigate the effects of cisapride on intestinal bacterial overgrowth (IBO), bacterial and endotoxin translocation, intestinal transit and permeability in cirrhotic rats.METHODS: All animals were assessed with variables including bacterial and endotoxin translocation, intestinal bacterial overgrowth, intestinal transit and permeability.Bacterial translocation (BT) was assessed by bacterial culture of MLN, liver and spleen, IBO by a jejunal bacterial count of the specific organism, intestinal permeability by determination of the 24-hour urinary 99mTc-DTPA excretion and intestinal transit by measurement of the distribution of 51Cr in the intestine.RESULTS: Bacterial translocation (BT) and IBO was found in 48 % and 80 % cirrhotic rats respectively and none in control rats. Urinary excretion of 99mTc-DTPA in cirrhotic rats with BT (22.2±7.8) was greater than these without BT (10.5±2.9). Intestinal transit (geometric center ratio) was significantly delayed in cirrhotic rats (0.31±0.06) and further more delayed in cirrhotic rats with BT (0.24±0.06) than these without BT (0.38±0.11). Cirrhotic rats with IBO had significantly higher rates of intestinal bacterial and endotoxin translocation, slower intestinal transit time and higher intestinal permeability than those without IBO. It was also found that BT was closely associated with IBO and the injury of intestinal barrier. Compared with the placebo group,cisapride-treated rats had lower rates of bacterial/endotoxin translocation and IBO, which was closely associated with increased intestinal transit and improved intestinal permeability by cisapride.CONCLUSION: These results indicate that endotoxin and bacterial translocation in cirrhotic rats may be attributed to IBO and increased intestinal permeability. Cisapride that accelerates intestinal transit and improve intestinal permeability might be helpful in preventing intestinal bacterial and endotoxin translocation.

  12. Effects of immune supplementation and immune challenge on bacterial assemblages in the avian cloaca

    NARCIS (Netherlands)

    Matson, Kevin D.; Versteegh, Maaike A.; van der Velde, Marco; Tieleman, B. Irene

    2015-01-01

    Relationships between avian physiology and bacterial assemblages in the cloaca are poorly understood. We used molecular techniques to analyze cloacal swabs from pigeons that were subjected to two immunological manipulations: lysozyme supplementation and endotoxin challenge. From the swabs, we derive

  13. Alcohol, intestinal bacterial growth, intestinal permeability to endotoxin, and medical consequences: summary of a symposium.

    Science.gov (United States)

    Purohit, Vishnudutt; Bode, J Christian; Bode, Christiane; Brenner, David A; Choudhry, Mashkoor A; Hamilton, Frank; Kang, Y James; Keshavarzian, Ali; Rao, Radhakrishna; Sartor, R Balfour; Swanson, Christine; Turner, Jerrold R

    2008-08-01

    This report is a summary of the symposium on Alcohol, Intestinal Bacterial Growth, Intestinal Permeability to Endotoxin, and Medical Consequences, organized by National Institute on Alcohol Abuse and Alcoholism, Office of Dietary Supplements, and National Institute of Diabetes and Digestive and Kidney Diseases of National Institutes of Health in Rockville, Maryland, October 11, 2006. Alcohol exposure can promote the growth of Gram-negative bacteria in the intestine, which may result in accumulation of endotoxin. In addition, alcohol metabolism by Gram-negative bacteria and intestinal epithelial cells can result in accumulation of acetaldehyde, which in turn can increase intestinal permeability to endotoxin by increasing tyrosine phosphorylation of tight junction and adherens junction proteins. Alcohol-induced generation of nitric oxide may also contribute to increased permeability to endotoxin by reacting with tubulin, which may cause damage to microtubule cytoskeleton and subsequent disruption of intestinal barrier function. Increased intestinal permeability can lead to increased transfer of endotoxin from the intestine to the liver and general circulation where endotoxin may trigger inflammatory changes in the liver and other organs. Alcohol may also increase intestinal permeability to peptidoglycan, which can initiate inflammatory response in liver and other organs. In addition, acute alcohol exposure may potentiate the effect of burn injury on intestinal bacterial growth and permeability. Decreasing the number of Gram-negative bacteria in the intestine can result in decreased production of endotoxin as well as acetaldehyde which is expected to decrease intestinal permeability to endotoxin. In addition, intestinal permeability may be preserved by administering epidermal growth factor, l-glutamine, oats supplementation, or zinc, thereby preventing the transfer of endotoxin to the general circulation. Thus reducing the number of intestinal Gram-negative bacteria

  14. Determination of bacterial endotoxin (pyrogen) in radiopharmaceuticals by the gel clot method. Validation

    International Nuclear Information System (INIS)

    Before the Limulus amebocyte lysate (LAL) test, the only available means of pirogenicity testing for parenteral drugs and medical devices was the United States Pharmacopoeia (USP) rabbit pyrogen test. Especially for radiopharmaceuticals, the LAL assay is the elective way to determine bacterial endotoxin. The aim of this work was to validate the gel clot method for some radiopharmaceuticals without measurable interference. The FDA's LALTest guideline defines interference as a condition that causes a significant difference between the endpoints of a positive water control and positive product control series using a standard endotoxin. Experiments were performed in accordance to the USP bacterial endotoxins test in the 131I- m-iodobenzylguanidine; the radioisotopes Gallium-67 and Thallium-201; the lyophilized reagents DTPA, Phytate, GHA, HSA and Colloidal Tin. The Maximum Valid Dilution (MVD) was calculated for each product based upon the clinical dose of the material and a twofold serial dilution below the MVD was performed in duplicate to detect interferences. The labeled sensitivity of the used LAL reagent was 0.125 EU mL-1 (Endotoxin Units per milliliter). For validation, a dilution series was performed, a twofold dilution of control standard endotoxin (CSE) from 0.5 to 0.03 EU mL-1, to confirm the labeled sensitivity of the LAL reagent being tested in sterile and non pyrogenic water, in quadruplicate. The same dilution series was performed with the CSE and the product in the 1:100 dilution factor, in three consecutive batches of each radiopharmaceutical. The products 131I-m-iodobenzylguanidine, Gallium-67, Thallium-201, DTPA, HSA and Colloidal Tin were found compatible with the LAL test at a 1:100 dilution factor. Phytate and GHA showed some interference in the gel clot test. Other techniques to determine endotoxins as the chromogenic (color development) and the turbidimetric test (turbidity development), were also assessed to get valuable quantitative and

  15. Effects of lactulose on intestinal endotoxin and bacterial translocation in cirrhotic rats

    Institute of Scientific and Technical Information of China (English)

    张顺财; 王唯; 任卫英; 戴茜; 贺伯明; 周康

    2003-01-01

    Objective To investigate the effects of lactulose on intestinal bacterial overgrowth (IBO), bacterial translocation (BT), intestinal transit and permeability in cirrhotic rats. Methods BT in all animals was assessed by bacterial culture of mesenteric lymph node (MLN), liver and spleen, and IBO was assessed by a jejunal bacterial count of the specific organism. Intestinal permeability was determined by the 24-hour urinary 99mTc-diethylenetriaminepentaacetate (99mTc-DTPA) excretion, and intestinal transit was determined by measuring the distribution of 51 Cr in the intestine. Results BT and IBO were found in 48% and 80% of the cirrhotic rats, respectively, while not in the control rats. Cirrhotic rats with IBO had significantly higher levels of intestinal endotoxin higher rates of bacterial translocation, shorter intestinal transit time and higher intestinal permeability than those without IBO. It was also found that BT was closely associated with IBO and injury of the intestinal barrier. Compared with the placebo group, lactulose-treated rats had lower rates of BT and IBO, which was closely associated with increased intestinal transit and improved intestinal permeability by lactulose. Conclusions Our study indicate that endotoxin and bacterial translocation in cirrhotic rats may attribute to IBO and increased intestinal permeability. Lactulose that accelerates intestinal transit and improves intestinal permeability might be helpful in preventing intestinal bacterial and endotoxin translocation.

  16. Control of dialysis water by microbial contamination and bacterial endotoxin tests

    Directory of Open Access Journals (Sweden)

    Fesem BAŞARI

    2012-01-01

    Full Text Available Objective: The aim of this study was to evaluate the microbial contamination and bacterial endotoxin levels in dialysis water samples sent to the Water and Food Microbiology Laboratory of the Adana Hygiene Institute Refik Saydam Hygiene Center, and to assess factors that might influence the results of these tests.Method: Two hundred fourty five dialysis water samples sent to the laboratory between January 2009 and December 2010 were tested; all of them for microbial contamination and 198 of them for bacterial endotoxin levels. Microbial contamination was evaluated by using the Plate Count Agar (PCA method while the Limulus Amoebocyte Lysate (LAL assay was used for the detection of bacterial endotoxins. The results were evaluated according to the criteria of the “Directives on the Water Purification Systems” and “the European Pharmacopoeia”. To compare the annual differences, the chi-square test was used for statistical evaluation and results with p0.05in 2010. On the other hand, in 26.7% of the dialysiswater samples the levels of bacterial endotoxinswas higher than 0.25 İU/ml in 2009, this percentagedropped to 16.5% in 2010 (p>0.05. During 2009-2010,245 microbial contamination tests were performedand in 20 (8.2% of the samples the TAMC values werehigher than 100 CFU/ml, while in 43 (21.7% out of198 of the samples the bacterial endotoxin levelswere higher than 0.25 İU/ml. Out of 20 in 43 samples,a bacterial endotoxin levels were >0.25 İU/ml werealso found as microbiologically contaminated ( >100CFU/ml, while in remaining 23 samples the level was<100 CFU/ml. There were no statistically significantdifferences in the percentages of positives found in2009 and 2010.Conclusion: The results of our study indicatethat the detection of endotoxins is a complementaryfactor to microbial contamination in the bacteriologicalmonitoring of dialysis waters. We recommend thatlevel of endotoxin should be taken into accountwhen the bacterial contamination

  17. Detection and Identification System of Bacteria and Bacterial Endotoxin Based on Raman Spectroscopy

    Directory of Open Access Journals (Sweden)

    Muhammad Elsayeh

    2016-03-01

    Full Text Available Sepsis is a global health problem that causes risk of death. In the developing world, about 60 to 80 % of death cases are caused by Sepsis. Rapid methods for detecting its causes, represent one of the major factors that may reduce Sepsis risks. Such methods can provide microbial detection and identification which is critical to determine the right treatment for the patient. Microbial and Pyrogen detection is important for quality control system to ensure the absence of pathogens and Pyrogens in the manufacturing of both medical and food products. Raman spectroscopes represent a q uick and accurate identification and detection method, for bacteria and bacterial endotoxin, which this plays an important role in delivering high quality biomedical products using the power of Raman spectroscopy. It is a rapid method for chemical structure detection that can be used in identifying and classifying bacteria and bacterial endotoxin. Such a method acts as a solution for time and cost effective quality control procedures. This work presents an automatic system based on Raman spectroscopy to detect and identify bacteria and bacterial endotoxin. It uses the frequency properties of Raman scattering through the interaction between organic materials and electromagnetic waves. The scattered intensities are measured and wave number converted into frequency, then the cepstral coefficients are extracted for both the detection and identification. The methodology depends on normalization of Fourier transformed cepstral signal to extract their classification features. Experiments’ results proved effective identification and detection of bacteria and bacterial endotoxin even with concentrations as low as 0.0003 Endotoxin unit (EU/ml and 1 Colony Forming Unit (CFU/ml using signal processing based enhancement technique.

  18. Invited review: Role of bacterial endotoxins in the etiopathogenesis of periparturient diseases of transition dairy cows.

    Science.gov (United States)

    Eckel, Emily F; Ametaj, Burim N

    2016-08-01

    The dairy industry continues to suffer severe economic losses due to the increased disease incidence cows experience during the transition period. It has long been the classical view that the major contributing factor to the development of these periparturient diseases is the considerable increase in nutritional demands for milk production. This classical view, however, fails to account for the substantial correlation between both metabolic and infectious diseases and the detrimental effects that can occur with the provision of high-energy diets to support these nutritional demands. Currently, increasing evidence implicates bacterial endotoxins in the etiopathology of most periparturient diseases. Bacterial endotoxins are components of the outer cell wall of gram-negative and gram-positive bacteria that are highly immunostimulatory and can trigger proinflammatory immune responses. The ability of endotoxins to translocate from the mucosal tissues, including the gastrointestinal tract, mammary gland, and uterus, into the systemic circulation has been observed. Once they have entered the circulation, endotoxins potentially contribute to disease either directly, through eliciting an inflammatory response, or indirectly through other factors such as the overreaction of the natural protective mechanisms of the host. Although the evidence implicating a role of endotoxins in the pathogenesis of transition diseases continues to grow, our current knowledge of the host response to mucosal endotoxin exposure and pathogenic mechanisms remain largely unknown. Developing our understanding of the connection between endotoxemia and dairy cattle disease holds significant potential for the future development of preventative measures that could benefit the productivity of the dairy industry as well as animal welfare. PMID:27209132

  19. Innate immune defenses exhibit circadian rhythmicity and differential temporal sensitivity to a bacterial endotoxin in Nile tilapia (Oreochromis niloticus).

    Science.gov (United States)

    Lazado, Carlo C; Skov, Peter Vilhelm; Pedersen, Per Bovbjerg

    2016-08-01

    was administered at ZT3. On the other hand, plasma melatonin was significantly affected by LPS injection but only when 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 is likely mediated by the time of day, highlighting the importance of circadian rhythm in the immunological functions of fish.

  20. Innate immune defenses exhibit circadian rhythmicity and differential temporal sensitivity to a bacterial endotoxin in Nile tilapia (Oreochromis niloticus).

    Science.gov (United States)

    Lazado, Carlo C; Skov, Peter Vilhelm; Pedersen, Per Bovbjerg

    2016-08-01

    was administered at ZT3. On the other hand, plasma melatonin was significantly affected by LPS injection but only when 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 is likely mediated by the time of day, highlighting the importance of circadian rhythm in the immunological functions of fish. PMID:27346154

  1. 脑复康注射液细菌内毒素检查法的探讨%Srudy on the Bacterial Endotoxin Determination of Ciclofalina Injection

    Institute of Scientific and Technical Information of China (English)

    王小虹; 刘卫卫; 王艶萍; 修锐

    2002-01-01

    Objective To establish the bacterial endotoxin detemination of ciclofalina injection. Methods The cxperiment was implemented according to the request of bacterial endotoxin determination of metronidazole collected by the 2ndbook, CHINA Pharmacopiea, 2000. Results The injection had no interference to the test when it was diluted for 1 times. Thedetecting results of the rabit test was compared with that of the bacterial endotoxin test. Conclusion Amebocye tysele methodcan be used as daily hopsital pyogen test of ciclofalina injection.

  2. 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

  3. 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

    The present study investigated the daily dynamics of humoral immune defenses and the temporal influence in the sensitivity of these responses to a bacterial endotoxin in Nile tilapia (Oreochromis niloticus). The first experiment subjected the fish to two photoperiod conditions, 12L:12D (LD) and 0L...

  4. Exposure to bacterial endotoxin generates a distinct strain of α-synuclein fibril.

    Science.gov (United States)

    Kim, Changyoun; Lv, Guohua; Lee, Jun Sung; Jung, Byung Chul; Masuda-Suzukake, Masami; Hong, Chul-Suk; Valera, Elvira; Lee, He-Jin; Paik, Seung R; Hasegawa, Masato; Masliah, Eliezer; Eliezer, David; Lee, Seung-Jae

    2016-01-01

    A single amyloidogenic protein is implicated in multiple neurological diseases and capable of generating a number of aggregate "strains" with distinct structures. Among the amyloidogenic proteins, α-synuclein generates multiple patterns of proteinopathies in a group of diseases, such as Parkinson disease (PD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA). However, the link between specific conformations and distinct pathologies, the key concept of the strain hypothesis, remains elusive. Here we show that in the presence of bacterial endotoxin, lipopolysaccharide (LPS), α-synuclein generated a self-renewable, structurally distinct fibril strain that consistently induced specific patterns of synucleinopathies in mice. These results suggest that amyloid fibrils with self-renewable structures cause distinct types of proteinopathies despite the identical primary structure and that exposure to exogenous pathogens may contribute to the diversity of synucleinopathies. PMID:27488222

  5. Addressing endotoxin issues in bioengineered heparin.

    Science.gov (United States)

    Suwan, Jiraporn; Torelli, Amanda; Onishi, Akihiro; Dordick, Jonathan S; Linhardt, Robert J

    2012-01-01

    Heparin is a widely used clinical anticoagulant that is prepared from pig intestine. A contamination of heparin in 2008 has led to a reexamination of animal-derived pharmaceuticals. A bioengineered heparin prepared by bacterial fermentation and chemical and enzymatic processing is currently under development. This study examines the challenges of reducing or removing endotoxins associated with this process that are necessary to proceed with preclinical in vivo evaluation of bioengineered heparin. The current process is assessed for endotoxin levels, and strategies are examined for endotoxin removal from polysaccharides and enzymes involved in this process. PMID:23586950

  6. Application of quartz tuning forks for detection of endotoxins and Gram-negative bacterial cells by monitoring of Limulus Amebocyte Lysate coagulation.

    Science.gov (United States)

    Chałupniak, Andrzej; Waszczuk, Karol; Hałubek-Głuchowska, Katarzyna; Piasecki, Tomasz; Gotszalk, Teodor; Rybka, Jacek

    2014-08-15

    Endotoxins, pyrogens of bacterial origin, are a significant threat in many areas of life. Currently, the test most commonly used for endotoxin level determination is LAL (Limulus Amebocyte Lysate) assay. This paper presents application of commercially available low-frequency piezoelectric tuning forks (QTFs) for endotoxin detection. Measurement of the decrease in the QTF oscillation amplitude provides information about the viscosity changes, occurring in the tested sample upon addition of LAL. That method was used to determine the concentrations of endotoxins and bacterial cells (E. coli O157:H19). The relevance of the obtained results was confirmed using a commercially available colorimetric LAL assay. The constructed system can detect bacterial endotoxins in the range of 0.001-5EU/ml and bacterial cells in the range of 10(2)-10(7)CFU/ml. The presented technique requires very simple sample preparation and the sensor response is obtained using compact, portable readout electronics. The single test cost is low compared to commercial endotoxin assays and other novel systems based on micromechanical sensors. PMID:24632139

  7. Establishment of Bacterial Endotoxins Test of Xiyanping Injection%喜炎平注射液细菌内毒素检查法的建立

    Institute of Scientific and Technical Information of China (English)

    吴寒寅; 孟德胜

    2012-01-01

    目的 建立喜炎平注射液细菌内毒素检查法.方法 按2010年版《中国药典(二部)》附录细菌内毒素检查法,用不同厂家的鲎试剂对3个批号的喜炎平注射液分别进行干扰试验和细菌内毒素检查.结果 喜炎平注射液稀释到40倍(质量浓度为1.25 g/L)时对细菌内毒素检查法无干扰,细菌内毒素限值L=5 EU/mL,按照拟订标准,3批样品细菌内毒素检查均符合规定.结论 所建立的鲎试剂法可用于喜炎平注射液的细菌内毒素检查.%Objective To establish the method of bacterial endotoxin test of Xiyanping injection. Methods Based on the bacterial endo-toxin test in the appendix of the Chinese Pharmacopeia(edition 2010,part 2),the interference test and the bacterial endotoxin test were conducted on the samples from 3 batches of Xiyanping injection by using tachypleus amebocyte lysate(TAL) from different manufacturers. Results Xiyanping injection solution with dilution to 40 times (mass concentration 1. 25 g/L) had no interference on the bacterial endotoxin test. The limit value of the bacterial endotoxin was 5 EU/mL. According to the established standard, the bacterial endotoxin test in the 3 batches of samples conformed to the stipulation. Conclusion The established TAL method may be applicable for the bacterial endotoxin test of Xiyanping injection.

  8. 叔丁醇细菌内毒素检查方法的建立%Establishment of Bacterial Endotoxins Test for Tertiary Butyl Alcohol

    Institute of Scientific and Technical Information of China (English)

    于海洲; 都婧; 于风平

    2015-01-01

    目的:建立叔丁醇细菌内毒素检查方法。方法:采用2个不同厂家的鲎试剂和3批样品,进行预干扰试验和干扰试验。结果:供试品溶液对鲎试剂与细菌内毒素之间的凝集反应无干扰;样品的细菌内毒素限值确定为0.25 EU·ml-1。结论:所建立的方法可行,可用于叔丁醇的细菌内毒素检查。%Objective:To establish a method for the detection of bacterial endotoxins in tertiary butyl alcohol. Methods:The in-terference experiment and bacterial endotoxins test for 3 batches of the products from 2 companies were carried out with tachypleus ame-bocyte lysate. Results:There was no interference in the bacterial endotoxins test. The limit of bacterial endotoxins was 0. 25 EU/ml. Conclusion:The established method of bacterial endotoxins test is feasible for tertiary butyl alcohol.

  9. Early weaning alters the acute-phase reaction to an endotoxin challenge in beef calves.

    Science.gov (United States)

    Carroll, J A; Arthington, J D; Chase, C C

    2009-12-01

    Previous research indicates that early weaning before shipment can reduce transportation-induced increases in acute-phase proteins (APP) and can increase feedlot performance in beef calves. These data suggest that the combination of weaning and transport stress may compromise the immune system of calves, thus hindering subsequent performance and health. Therefore, our objective was to determine if the innate immune response of early weaned calves (EW; 80 d of age) differed from normal-weaned calves (NW; 250 d of age) in response to an endotoxin challenge. Eighteen Brahman x Angus calves (8 and 10 EW and NW, respectively; 233 +/- 5 kg of BW) were used. Calves were maintained on pasture with supplement and then moved into individual pens for 1 wk of acclimation before the start of the study. Calves were fitted with an indwelling jugular catheter 1 d before LPS challenge (0 h; 1.0 microg/kg of BW, intravenously). Blood samples were collected at 30-min intervals from -2 to 8 h. Serum samples were stored at -80 degrees C until analyzed for cortisol, tumor necrosis factor-alpha (TNF), IL-1 beta, IL-6, interferon-gamma (IFN), ceruloplasmin, and haptoglobin. Whereas LPS increased serum cortisol (P or= 0.15) was observed. A weaning age x time interaction (P x time interaction (P challenge compared with that of NW calves. Additionally, the differential IFN response indicates that the immune system of EW calves may be more effective at recognizing and eliminating endotoxin. These data suggest that an altered innate immune system may be one of the factors responsible for the improved feedlot performance previously reported in EW calves. PMID:19717781

  10. Microbiological assessment of house and imported bottled water by comparison of bacterial endotoxin concentration, heterotrophic plate count, and fecal coliform count.

    Science.gov (United States)

    Reyes, Mayra I; Pérez, Cynthia M; Negrón, Edna L

    2008-03-01

    Consumers increasingly use bottled water and home water treatment systems to avoid direct tap water. According to the International Bottled Water Association (IBWA), an industry trade group, 5 billion gallons of bottled water were consumed by North Americans in 2001. The principal aim of this study was to assess the microbial quality of in-house and imported bottled water for human consumption, by measurement and comparison of the concentration of bacterial endotoxin and standard cultivable methods of indicator microorganisms, specifically, heterotrophic and fecal coliform plate counts. A total of 21 brands of commercial bottled water, consisting of 10 imported and 11 in-house brands, selected at random from 96 brands that are consumed in Puerto Rico, were tested at three different time intervals. The Standard Limulus Amebocyte Lysate test, gel clot method, was used to measure the endotoxin concentrations. The minimum endotoxin concentration in 63 water samples was less than 0.0625 EU/mL, while the maximum was 32 EU/mL. The minimum bacterial count showed no growth, while the maximum was 7,500 CFU/mL. Bacterial isolates like P. fluorescens, Corynebacterium sp. J-K, S. paucimobilis, P. versicularis, A. baumannii, P. chlororaphis, F. indologenes, A. faecalis and P. cepacia were identified. Repeated measures analysis of variance demonstrated that endotoxin concentration did not change over time, while there was a statistically significant (p count over time. In addition, multiple linear regression analysis demonstrated that a unit change in the concentration of endotoxin across time was associated with a significant (p count. This analysis evidenced a significant time effect in the average log bacteriological cell count. Although bacterial growth was not detected in some water samples, endotoxin was present. Measurement of Gram-negative bacterial endotoxins is one of the methods that have been suggested as a rapid way of determining bacteriological water quality.

  11. Lung responses to secondary endotoxin challenge in rats exposed to pig barn air

    Directory of Open Access Journals (Sweden)

    Townsend Hugh GG

    2008-10-01

    Full Text Available Abstract Background Swine barn air contains endotoxin and many other noxious agents. Single or multiple exposures to pig barn air induces lung inflammation and loss of lung function. However, we do not know the effect of exposure to pig barn air on inflammatory response in the lungs following a secondary infection. Therefore, we tested a hypothesis that single or multiple exposures to barn air will result in exaggerated lung inflammation in response to a secondary insult with Escherichia coli LPS (E. coli LPS. Methods We exposed Sprague-Dawley rats to ambient (N = 12 or swine barn air (N = 24 for one or five days and then half (N = 6/group of these rats received intravenous E. coli LPS challenge, observed for six hours and then euthanized to collect lung tissues for histology, immunohistochemistry and ELISA to assess lung inflammation. Results Compared to controls, histological signs of lung inflammation were evident in barn exposed rat lungs. Rats exposed to barn air for one or five days and challenged with E. coli LPS showed increased recruitment of granulocytes compared to those exposed only to the barn. Control, one and five day barn exposed rats that were challenged with E. coli LPS showed higher levels of IL-1β in the lungs compared to respective groups not challenged with E. coli LPS. The levels of TNF-α in the lungs did not differ among any of the groups. Control rats without E. coli LPS challenge showed higher levels of TGF-β2 compared to controls challenged with E. coli LPS. Conclusion These results show that lungs of rats exposed to pig barn air retain the ability to respond to E. coli LPS challenge.

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

    Directory of Open Access Journals (Sweden)

    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

  13. 注射用丹参多酚酸盐细菌内毒素检查%Feasibility Study on Bacterial Endotoxin Test of Salvianolate for Injection

    Institute of Scientific and Technical Information of China (English)

    汪伟

    2012-01-01

    Objective: To establish a method for the bacterial endotoxin test of salvianolate for injection. Method: The bacterial endotoxin test was carried out according to the requirements in the appendix XI E of Chinese Pharmacopeia( edition 2010, Vol II ). Result: The salvianolate for injection at the concentration not greater than 0. 8 mg ? Ml-1 showed no interference on tachypiens amebocyte lysate teat. Conclusion: Bacterial endotoxin test ( gel process ) is applicable in the bacterial endotoxin limit test of salvianolate for injection.%目的:建立注射用丹参多酚酸盐的细菌内毒素检查方法.方法:按2010年版(二部)附录ⅪE细菌内毒素检查法进行操作.结果:注射用丹参多酚酸盐稀释至0.8 mg·ml-1及以下浓度对鲎试剂干扰试验无干扰作用.结论:注射用丹参多酚酸盐可采用细菌内毒素检查法(凝胶法)进行细菌内毒素的限量检查.

  14. 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.

  15. Study on Testing for Bacterial Endotoxin in Ibuprofen%布洛芬细菌内毒素检查法研究

    Institute of Scientific and Technical Information of China (English)

    张德波

    2012-01-01

    OBJECTIVE To establish a method of examining bacterial endotoxin for ibuprofen. METHODS To determine the reasonable limit of bacterial endotoxin by Chinese Pharmacopoeia 2010; and study on testing for bacterial endotoxin to determine the noninterference concentration for ibuprofen. RESULTS According the dose of clinical medication to determine the most suitable limit of bacterial endotoxin in preparations for ibuprofen was 0.20 EU·mg‐1 and uesd the TAL of two manufacturers to test, and it was proved that the maximum noninterference concentration was 0.625 mg·mL‐1 and the best testing concentration was no more than 0.625 mg·mL‐1. CONCLUSION It was showed that the method is suitable for the ibuprofen of bacterial endotoxin test.%目的 建立布洛芬细菌内毒素检查方法.方法 按中国药典2010年版二部,确定合理的布洛芬内毒素限值,并研究布洛芬对细菌内毒素与鲎试剂反应的干扰情况,以确定其最大不干扰试验浓度.结果 根据临床实际应用情况,确定布洛芬的内毒素限值为L=0.20 EU·mg-1,使用2个厂家生产的鲎试剂试验,结果 本品配制成≤0.625 mg·mL-1浓度对反应无干扰作用.结论 细菌内毒素检查法可用于布洛芬细菌内毒素的检查.

  16. Effects of coupled dose and rhythm manipulation of plasma cortisol levels on leukocyte transcriptional response to endotoxin challenge in humans.

    Science.gov (United States)

    Kamisoglu, Kubra; Sleight, Kirsten; Nguyen, Tung T; Calvano, Steve E; Coyle, Susette M; Corbett, Siobhan A; Androulakis, Ioannis P

    2014-10-01

    Severe traumas are associated with hypercortisolemia due to both disruption of cortisol secretion rhythm and increase in its total concentration. Understanding the effects of altered cortisol levels and rhythms on immune function is of great clinical interest, to prevent conditions such as sepsis from complicating the recovery. This in vivo study assesses the responses of circulating leukocytes to coupled dose and rhythm manipulation of cortisol, preceding an immune challenge induced by endotoxin administration. Through continuous infusion, plasma cortisol concentration was increased to and kept constant at a level associated with major physiologic stress. In response, transcriptional programming of leukocytes was altered to display a priming response before endotoxin exposure. Enhanced expression of a number of receptors and signaling proteins, as well as lowered protein translation and mitochondrial function indicated a sensitization against potential infectious threats. Despite these changes, response to endotoxin followed very similar patterns in both cortisol and saline pre-treated groups except one cluster including probe sets associated with major players regulating inflammatory response. In sum, altered dose and rhythm of plasma cortisol levels engendered priming of circulating leukocytes when preceded an immune challenge. This transcriptional program change associated with stimulated surveillance function and suppressed energy-intensive processes, emphasized permissive actions of cortisol on immune function.

  17. Endotoxin, capsule, and bacterial attachment contribute to Neisseria meningitidis resistance to the human antimicrobial peptide LL-37.

    Science.gov (United States)

    Jones, Allison; Geörg, Miriam; Maudsdotter, Lisa; Jonsson, Ann-Beth

    2009-06-01

    Pathogenic bacteria have evolved numerous mechanisms to evade the human immune system and have developed widespread resistance to traditional antibiotics. We studied the human pathogen Neisseria meningitidis and present evidence of novel mechanisms of resistance to the human antimicrobial peptide LL-37. We found that bacteria attached to host epithelial cells are resistant to 10 microM LL-37 whereas bacteria in solution or attached to plastic are killed, indicating that the cell microenvironment protects bacteria. The bacterial endotoxin lipooligosaccharide and the polysaccharide capsule contribute to LL-37 resistance, probably by preventing LL-37 from reaching the bacterial membrane, as more LL-37 reaches the bacterial membrane on both lipooligosaccharide-deficient and capsule-deficient mutants whereas both mutants are also more susceptible to LL-37 killing than the wild-type strain. N. meningitidis bacteria respond to sublethal doses of LL-37 and upregulate two of their capsule genes, siaC and siaD, which further results in upregulation of capsule biosynthesis.

  18. Bacterial challenge of NISSHO ultrafilter ETF 609: results of in vitro testing.

    Science.gov (United States)

    Krautzig, S; Lonnemann, G; Shaldon, S; Koch, K M

    1996-07-01

    In hemodialysis, a certain degree of bacterial contamination on the dialysate side is a regular finding. Concern has been growing that this contamination may lead to a chronic inflammatory response in the patient. Ultrafiltration of dialysate can be used to reduce bacterial content and levels of cytokine-inducing substances upstream of the patient's dialyzer. The aim of this study was to test in vitro the rejection capacity of a polysulfone hollow-fiber ultrafilter (ETF 609, NISSHO Co., Osaka, Japan) challenged with bacterial filtrates derived from Pseudomonas aeruginosa PA103. Results showed a reduction of interleukin-1 beta-inducing activity (measured on peripheral blood mononuclear cells) from 5,035 +/- 394 pg/ml prefilter to nondetectable levels postfilter and endotoxin levels (limulus amebocyte lysate assay) of 4,167 +/- 1,079 versus 12 +/- 2 pg/ml, respectively. In conclusion, ultrafiltration of dialysate with the polysulfone ultrafilter ETF 609 leads to a potent reduction of cytokine-inducing activity.

  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. Determination of bacterial endotoxin (pyrogen) in radiopharmaceuticals by the gel clot method. Validation; Determinacao de endotoxina bacteriana (pirogenio) em radiofarmacos pelo metodo de formacao de gel. Validacao

    Energy Technology Data Exchange (ETDEWEB)

    Fukumori, Neuza Taeko Okasaki

    2008-07-01

    Before the Limulus amebocyte lysate (LAL) test, the only available means of pirogenicity testing for parenteral drugs and medical devices was the United States Pharmacopoeia (USP) rabbit pyrogen test. Especially for radiopharmaceuticals, the LAL assay is the elective way to determine bacterial endotoxin. The aim of this work was to validate the gel clot method for some radiopharmaceuticals without measurable interference. The FDA's LALTest guideline defines interference as a condition that causes a significant difference between the endpoints of a positive water control and positive product control series using a standard endotoxin. Experiments were performed in accordance to the USP bacterial endotoxins test in the {sup 131}I- m-iodobenzylguanidine; the radioisotopes Gallium-67 and Thallium-201; the lyophilized reagents DTPA, Phytate, GHA, HSA and Colloidal Tin. The Maximum Valid Dilution (MVD) was calculated for each product based upon the clinical dose of the material and a twofold serial dilution below the MVD was performed in duplicate to detect interferences. The labeled sensitivity of the used LAL reagent was 0.125 EU mL{sup -1} (Endotoxin Units per milliliter). For validation, a dilution series was performed, a twofold dilution of control standard endotoxin (CSE) from 0.5 to 0.03 EU mL{sup -1}, to confirm the labeled sensitivity of the LAL reagent being tested in sterile and non pyrogenic water, in quadruplicate. The same dilution series was performed with the CSE and the product in the 1:100 dilution factor, in three consecutive batches of each radiopharmaceutical. The products {sup 131}I-m-iodobenzylguanidine, Gallium-67, Thallium-201, DTPA, HSA and Colloidal Tin were found compatible with the LAL test at a 1:100 dilution factor. Phytate and GHA showed some interference in the gel clot test. Other techniques to determine endotoxins as the chromogenic (color development) and the turbidimetric test (turbidity development), were also assessed to get

  1. Lung responses to secondary endotoxin challenge in rats exposed to pig barn air

    OpenAIRE

    Townsend Hugh GG; Aulakh Gurpreet K; Keet Taryn; Charavaryamath Chandrashekhar; Singh Baljit

    2008-01-01

    Abstract Background Swine barn air contains endotoxin and many other noxious agents. Single or multiple exposures to pig barn air induces lung inflammation and loss of lung function. However, we do not know the effect of exposure to pig barn air on inflammatory response in the lungs following a secondary infection. Therefore, we tested a hypothesis that single or multiple exposures to barn air will result in exaggerated lung inflammation in response to a secondary insult with Escherichia coli...

  2. TRPA1 channels mediate acute neurogenic inflammation and pain produced by bacterial endotoxins

    Science.gov (United States)

    Meseguer, Victor; Alpizar, Yeranddy A.; Luis, Enoch; Tajada, Sendoa; Denlinger, Bristol; Fajardo, Otto; Manenschijn, Jan-Albert; Fernández-Peña, Carlos; Talavera, Arturo; Kichko, Tatiana; Navia, Belén; Sánchez, Alicia; Señarís, Rosa; Reeh, Peter; Pérez-García, María Teresa; López-López, José Ramón; Voets, Thomas; Belmonte, Carlos; Talavera, Karel; Viana, Félix

    2014-01-01

    Gram-negative bacterial infections are accompanied by inflammation and somatic or visceral pain. These symptoms are generally attributed to sensitization of nociceptors by inflammatory mediators released by immune cells. Nociceptor sensitization during inflammation occurs through activation of the Toll-like receptor 4 (TLR4) signalling pathway by lipopolysaccharide (LPS), a toxic by-product of bacterial lysis. Here we show that LPS exerts fast, membrane delimited, excitatory actions via TRPA1, a transient receptor potential cation channel that is critical for transducing environmental irritant stimuli into nociceptor activity. Moreover, we find that pain and acute vascular reactions, including neurogenic inflammation (CGRP release) caused by LPS are primarily dependent on TRPA1 channel activation in nociceptive sensory neurons, and develop independently of TLR4 activation. The identification of TRPA1 as a molecular determinant of direct LPS effects on nociceptors offers new insights into the pathogenesis of pain and neurovascular responses during bacterial infections and opens novel avenues for their treatment.

  3. A rapid kinetic chromogenic method for quantification of bacterial endotoxins in lyophilized reagents for labeling with {sup 99m}Tc radiopharmaceuticals

    Energy Technology Data Exchange (ETDEWEB)

    Fukumori, Neuza T.O.; Campos, Domingos G.; Silva, Laercio; Fernandes, Adriana V.; Mengatti, Jair; Silva, Constancia P.G.; Matsuda, Margareth M.N. [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)

    2009-07-01

    A rapid quantitative kinetic chromogenic test in an automated Portable Test System (PTS) has been developed for determination of bacterial endotoxins in water, in-process and end-products using the Limulus amebocyte lysate (LAL). The aim of this work was to validate the method for lyophilized reagents for labeling with {sup 99m}Tc radiopharmaceuticals with no interfering factors. Experiments were performed in three consecutive batches of the lyophilized reagents Methylenediphosphonic Acid (MDP) and Pyrophosphate (PYRO) produced at IPEN-CNEN/ SP using the PTS from Endosafe, Inc.{sup TM}, Charleston, SC. The Maximum Valid Dilution (MVD) was calculated to establish the extent of dilution to avoid interfering test conditions (MVD=500). Better results were obtained above 1:20 dilution factor for MDP and 1:100 for PYRO. The parameters of coefficient correlation (R) -0.980, RPPC between 50 - 200% and coefficient variation (CV) of the samples less than 25% were satisfied and the endotoxin concentration was lower than the lowest concentration of the standard curve (0.05 EU mL{sup -1}), therefore less than the established limit in pharmacopoeias. The PTS is a rapid, simple and accurate technique using the quantitative kinetic chromogenic method for bacterial endotoxin determination. For this reason, it is very practical in the radiopharmaceutical area and it trends to be the method of choice for the pyrogen test. For MDP and PYRO, the validation was successfully performed. (author)

  4. Study on Determination of Bacterial Endotoxin of Ozagrel Injection%奥扎格雷注射液细菌内毒素检查法的研究

    Institute of Scientific and Technical Information of China (English)

    2013-01-01

      目的用细菌内毒素检查法检测奥扎格雷注射液中的热原,减少临床热原反应的发生。方法按照《中国药典》2010版(二部)附录细菌内毒素检查法和细菌内毒素检查指导原则,进行干扰实验,测定注奥扎格雷注射液细菌内毒素限定值。结果本品稀释16倍时对测定无干扰,限值为2.5 EU/mL。结论细菌内毒素法简便、快速、灵敏,能有效检测奥扎格雷注射液中热原。%Objective To detect the limulus in ozagrel injection with a bacterial endotoxin test. Method The interference test was performed according to the bacterial endotoxin method and its guiding principle assigned in "Chinese Pharmacopoeia 2010 version".Results The ozagrel injection showed no interference after diluted 16 times with a limit of 2.5 EU/mL. Conclusion Bacterinal Endotoxin method is simple, rapid, sensitive, and can effectively detect the pyrogen of ozagrel injection .

  5. Detection of bacterial endotoxin in Xingnaojing injection with Tachypleus Amebocyte lysate%鲎试剂检查醒脑静注射液细菌内毒素的探讨

    Institute of Scientific and Technical Information of China (English)

    叶树林; 王晓蕾

    2011-01-01

    Objective To study the method for the detection of bacterial endotoxin in Xingnaojing injection with Tachypleus Amebocyte lysate(TAL). Methods Experiments were performed according to the bacterial endotoxin tests covered in Chinese Pharmacopeia 2005(part 2). Results Xingnaojing injection did not interfere with its gel reaction to the bacterial endotoxin when it was diluted 10-fold. Conclusion Bacterial endotoxin test can be used to detect bacterial endotoxin in Xingnaojing injection.%目的 探讨用鲎试剂检查醒脑静注射液细菌内毒素方法.方法 根据2005年版Ⅱ部收载的细菌内毒素检查法的要求进行实验.结果 醒脑静注射液经10倍稀释时不干扰鲎试剂与细菌内毒素的凝胶反应.结论 细菌内毒素检查法适用于检测醒脑静注射液的内毒素.

  6. Rapid removal of bacterial endotoxin and natural organic matter in water by dielectric barrier discharge plasma: Efficiency and toxicity assessment.

    Science.gov (United States)

    Zhang, Can; Fang, Zhendong; Liu, Wenjun; Tian, Fang; Bai, Miao

    2016-11-15

    Low-temperature plasma was used to control bacteria, endotoxins and natural organic matter (NOM) in water by a dielectric barrier discharge (DBD) device. Results indicate that DBD plasma has an obvious inactivation effect on various bacteria in water. The degree of inactivation from difficult to easy is as follows: Bacillus subtilis>Escherichia coli>Staphylococcus aureus. Activated ultrapure water treated using DBD plasma exhibited a sustained sterilization effect, but this sterilization effect decreased gradually after 1h. The total-endotoxin (free-endotoxin and bound-endotoxin) released by Escherichia coli during inactivation, as well as artificially simulated endotoxin in a control solution, was significantly controlled by DBD plasma. Both the metabolites that appeared after inactivation of microorganisms by plasma treatment, and the NOM in filtration effluent of a water treatment plant were well removed by DBD plasma if the treatment duration was sufficiently long. However, the acute toxicity increased significantly, and persisted for at least 2h, indicating that some long-life active substances were generated during the DBD process. Therefore, the removal of bacteria, endotoxins or NOM does not mean a safe water is produced. It is also important to eliminate the toxicity and byproducts produced during water treatment for the continuous promotion and industrial application of DBD plasma. PMID:27388420

  7. Study on bacterial endotoxins test of disodium cantharidinate and vitamin B_6 injection%斑蝥酸钠维生素B_6注射液细菌内毒素检查法研究

    Institute of Scientific and Technical Information of China (English)

    王贵英; 唐继坤; 潘正兴

    2009-01-01

    Objective:To establish a method for determination of the bacterial endotoxin test in disodium cantharidinate and vitamin B_6 injection.Method:Inhibition and bacterial endotoxin test was performed with amebcyte lysate manufactured by different companies with different specifications according to the method of bacterial endotoxin test approved by Chinese Pharmacopoeia 2005 edition.Results:The original solution had non-interference with the bacterial endotoxin test.The results of bacteria endotoxin test in six batchs all accorded with the standard of the quality control.Conclusion:The results suggested that bacteria endotoxin test can be used as an alternative method for the rabbit pyrogen test for disodium cantharidinate and vitamin B_6 injection.The limits of bacterial endotoxin was 6 EU·mg~(-1).%目的:建立斑蝥酸钠维生素B6注射液的细菌内毒素检查方法.方法:按中国药典2005年版二部附录XIE细菌内毒素检查法进行,用小同厂家不同规格的鲎试剂对小同批号的斑蝥酸钠维牛素B_6注射液进行了干扰试验和细菌内毒素检查.结果:本品原液对细菌内毒素检查无干扰作用.按拟定标准检验,该品种6批样品细菌内毒素检查结果均符合规定.结论:本品可用细菌内毒素检查法代替家兔热原检查法,其细菌内毒素的限值(L)为6 EU·mL~(-1).

  8. Content Determination of Bacterial Endotoxin in Azithromycin by Kinetic Turbidimetric Limulus Method%动态浊度法测定阿奇霉素中细菌内毒素含量

    Institute of Scientific and Technical Information of China (English)

    张娟; 祝清芬; 范治云; 魏霞; 史国生

    2014-01-01

    目的:建立测定阿奇霉素中细菌内毒素含量的方法。方法采用动态浊度法,通过测定供试液中外加内毒素的回收率进行干扰试验,确定样品检测质量浓度线性范围,并定量测定样品中的细菌内毒素。结果内毒素检测质量浓度线性范围为0.03125~2.0 EU/mL ( r=-0.9989);样品在2.5 g/L及以下质量浓度时,对试验无干扰,细菌内毒素回收率在50%~200%范围内,样品中内毒素的含量可定量测定。结论动态浊度法可用于阿奇霉素中细菌内毒素的定量检测。%Objective To establish a method for determining the content of bacterial endotoxin in azithromycin. Methods The kinetic turbidimetric limulus method was adopted to generate the standard curve of bacterial endotoxin. The interference test was performed by determining the recovery rate of additional bacterial endotoxin in the test sample solution. Furthermore the bacterial endotoxin content in samples was quantitatively determined. Results The linear range of bacterial endotoxin detection concentration was 0. 031 52-2. 0 EU/mL ( r= -0. 998 9);the sample under the concentration ≤2. 5 g/L had no interference to test. The recovery rate of bacterial endotoxin was within the range of 50% -200%. The bacterial endotoxin content in the sample could be quantitatively determined. Conclusion The kinetic turbidimetric limulus method can be used to the quantitative determination of bacterial endotoxin in azithromycin.

  9. Endotoxin dosage in sepsis

    Directory of Open Access Journals (Sweden)

    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.

  10. 枸橼酸钠注射液细菌内毒素检查方法的考察%Studies of Bacterial Endotoxins Test of Sodium Citrate Injection

    Institute of Scientific and Technical Information of China (English)

    何进; 史国兵; 高军; 陈金旺

    2011-01-01

    Objective To establish a method of bacterial endotoxin test of sodium citrate injection by gel clot.Methods The method was based on the bacterial endotoxin test in the appendix of Chp 2005.A series of tests were performed, such as the sensitivity test of tachypleus amebocyte lysate, first interference test and interference tests.Results No interference was found after the sodium citrate injection was diluted 6 times in the bacterial endotoxin interference factors test.Conclusion The present method can replace the pyrogen test with rabbits for examination of bacterial endotoxin of sodium citrate injection.%目的 建立枸橼酸钠注射液细菌内毒素检查方法.方法 按2005年版附录细菌内毒素检查法,分别进行了鲎试剂灵敏度复核试验、细菌内毒素干扰试验.结果 枸橼酸钠注射液稀释6倍后,不干扰鲎试剂与细菌内毒素的凝胶反应.结论 可用细菌内毒素检查法替代家兔法检查枸橼酸钠注射液中的细菌内毒素.

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

    OpenAIRE

    Mario Roberto Leonardo; Raquel Assed Bezerra da Silva; Sada Assed; Paulo Nelson-Filho

    2004-01-01

    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 viru...

  12. Endotoxin detection--from limulus amebocyte lysate to recombinant factor C.

    Science.gov (United States)

    Ding, Jeak Ling; Ho, Bow

    2010-01-01

    Gram negative bacterial endotoxin is a biological pyrogen that causes fever when introduced intravenously. The endotoxin, also known as lipopolysaccharide (LPS), is found in the outer membrane of Gram-negative bacteria. During Gram-negative sepsis, endotoxin stimulates host macrophages to release inflammatory cytokines. However, excessive inflammation causes multiple organ failure and death. Endotoxins, which are ubiquitous pathogenic molecules, are a bane to the pharmaceutical industry and healthcare community. Thus early and sensitive detection of endotoxin is crucial to prevent endotoxaemia. The limulus amebocyte lysate (LAL) has been widely used for ~30 years for the detection of endotoxin in the quality assurance of injectable drugs and medical devices. The LAL constitutes a cascade of serine proteases which are triggered by trace levels of endotoxin, culminating in a gel clot at the end of the reaction. The Factor C, which normally exists as a zymogen, is the primer of this coagulation cascade. In vivo, Factor C is the perfect biosensor, which alerts the horseshoe crab of the presence of a Gram-negative invader. The hemostatic end-point entraps the invader, killing it and limiting further infection. However, as an in vitro endotoxin detection tool, variations in the sensitivity and specificity of LAL to endotoxin, and the dwindling supply of horseshoe crabs are posing increasing challenges to the biotechnology industry. This has necessitated the innovation of an alternative test for endotoxin. Thus, Factor C became the obvious, albeit tricky target for the recombinant technology effort. This chapter documents the backwater of mining the natural blood lysate of the endangered species to the monumental effort of genetic engineering, to produce recombinant Factor C (rFC). The rFC is a 132 kDa molecule, which was produced as a proenzyme inducible by the presence of trace levels of endotoxin. The rFC forms the basis of the "PyroGene" kit, which is a novel micro

  13. 注射用维生素C中细菌内毒素的定量检测%Study on quantitative determination of bacterial endotoxins in vitamin C for injection

    Institute of Scientific and Technical Information of China (English)

    汤艳群; 肖贵南

    2014-01-01

    Objective To establish a method for quantitative determination of bacterial endotoxins in vitamin C for injection. Methods Kinetic turbidimetric limulus test was applied to detect the bacterial endotoxins in vitamin C for injection quantitatively, and the result was compared with that of gel-clot method. Results Vitamin C for injection obviously interfered with the test for bacterial endotoxins when diluted to the concentration of 8 mg·mL-1,but not at 4 mg·mL-1 or below;the content of bacterial endotoxins in all samples tested were less than 0.02 EU·mg -1 ,which were in accordance with the result of gel-clot method. Conclusion Kinetic turbidimetric limulus test could be applied to determine bacterial endotoxins in vitamin C for injection.%目的:建立定量检测注射用维生素C中细菌内毒素的方法。方法采用动态浊度法鲎试验对样品中的细菌内毒素进行检测,并与凝胶法测定结果相比较。结果注射用维生素C在稀释至8 mg·mL-1时对细菌内毒素检查有明显的干扰作用,而在4 mg · mL-1或以下质量浓度时则无干扰作用;所测样品中细菌内毒素含量均低于0.02 EU·mg-1,与凝胶法测定结果一致。结论动态浊度法鲎试验可应用于定量检测注射用维生素C中的细菌内毒素。

  14. 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. PMID:23934306

  15. Involvement of serotonergic pathways in mediating the neuronal activity and genetic transcription of neuroendocrine corticotropin-releasing factor in the brain of systemically endotoxin-challenged rats

    Energy Technology Data Exchange (ETDEWEB)

    Laflamme, N.; Feuvrier, E.; Richard, D.; Rivest, S. [Laboratory of Molecular Endocrinology, CHUL Research Center and Department of Anatomy and Physiology, Laval University, 2705 boul. Laurier, Ste-Foy Quebec (Canada)

    1999-01-01

    The present study investigated the effect of serotonin depletion on the neuronal activity and transcription of corticotropin-releasing factor in the rat brain during the acute-phase response. Conscious male rats received an intraperitoneal (i.p.) injection with the immune activator lipopolysaccaride (25 {mu}g/100 g body wt) after being treated for three consecutive days with para-chlorophenylalanine (30 mg/100 g/day). This irreversible inhibitor of tryptophane-5-hydroxylase decreased hypothalamic serotonin levels by 96%. One, 3 and 6 h after a single i.p. injection of lipopolysaccharide or vehicle solution, rats were killed and their brains cut in 30-{mu}m coronal sections. Messenger RNAs encoding c-fos, nerve-growth factor inducible-B gene, corticotropin-releasing factor and the heteronuclear RNA encoding corticotropin-releasing factor primary transcript were assayed by in situ hybridization using {sup 35}S-labeled riboprobes, whereas Fos-immunoreactive nuclei were labeled by immunocytochemistry. Lipopolysaccharide induced a wide neuronal activation indicated by the expression of both immediate-early gene transcripts and Fos protein in numerous structures of the brain. The signal for both immediate-early gene transcripts was low to moderate 1 h after lipopolysaccharide administration, maximal at 3 h and decline at 6 h post-injection, whereas at that time, Fos-immunoreactive nuclei were still detected in most of the c-fos messenger RNA-positive structures. Interestingly, the strong and widespread induction of both immediate-early gene transcripts was almost totally inhibited by para-chlorophenylalanine treatment; in the hypothalamic paraventricular nucleus for example, c-fos messenger RNA signal and the number of Fos-immunoreactive positive cells were reduced by 80 and 48%, respectively, in serotonin-depleted rats treated with the bacterial endotoxin. This blunted neuronal response was also associated with an attenuated stimulation of neuroendocrine corticotropin

  16. Quantitative method for the Measurement of bacterial endotoxin in Hirudo injection%动态浊度法检测水蛭注射液中细菌内毒素

    Institute of Scientific and Technical Information of China (English)

    肖伟

    2012-01-01

      Objective to develop an assay for determination of bacterial endotoxin in Hirudo injection and compare it with pyrogens test. Methods detection was carried out according to the bacterial endotoxin test procedure and guiding principle for interfere factors test in chinese Pharmacopeia,2005 edition. Rsults Hirudo injection did not interfere with limulus agent in 200 diluted concentration,it can get the same result which meets Pharmacopeia,2005 edition using the two methods. Conclusion turbidimetric-Kinetic method for the Measurement of bacterial endotoxin could be used in in Hirudo injection.%  目的建立动态浊度法定量检测水蛭注射液中的细菌内毒素。方法按中国药典2010版二部收载的细菌内毒素检查方法及其指导原则进行干扰试验,并与热源检查法比较。结果当样品作200倍稀释时用动态浊度法进行测试未见干扰作用,与热源法测试结果一致,均符合规定。结论该样品可以用动态浊度法进行定量检测水蛭注射液中的内毒素。

  17. 盐酸艾司洛尔注射液细菌内毒素检查法的实验研究%The Experiment of Bacterial Endotoxin Test for Esmolol Hydrochloride Injection

    Institute of Scientific and Technical Information of China (English)

    周修森

    2011-01-01

    Objective To establish the bacterial endotoxins determination method for esmolol hydrochloride injection.Methods According to the method in the appendix of the Chinese Pharmacopoeia(edition 2010), the bacterial endotoxin test was carried out. Results The maximum non-disturb concentration of esmolol hydrochloride is 1.2mg·mL-1.Conclusion The method of bacterial endotoxin test is feasible with the limit of 0.20 EU·mg-1 for esmolol hydrochlo ride injection.%目的 建立盐酸艾司洛尔注射液的细菌内毒素检查法.方法 按2010年版细菌内毒素检查法相关规定进行试验.结果 本品最大不干扰浓度以盐酸艾司洛尔计为1.2mg·mL-1.结论 所建立的盐酸艾司洛尔注射液的细菌内毒素检查方法具有可行性,其内毒素限值可定为0.20EU·mg-1.

  18. Determination of Bacterial Endotoxin in Xueshuantong Injecta by Kinetic Nephelometry%动态浊度法测定血栓通注射液细菌内毒素含量

    Institute of Scientific and Technical Information of China (English)

    巴力吉尼玛

    2012-01-01

    Objective:To establish a kinetic nephelimetry method for the determination of bacterial endotoxin in Xueshuantong Injecta. Methods:In accordance with the Bacterial Endotoxin test in China Pharmacopoeia (2010,Volume Ⅱ )the samples were diluted respectively from 25 to 200 folds.And then the interference for Limulus test was eliminated effectively. Results and Conclusion:The kinetic nephelometric assay can be used for quantitative determination of bacterial endotoxin in Xueshantong Injecta.%目的:建立血栓通注射液中细菌内毒素定量分析方法.方法:参照《中国药典》2010年版二部细菌内毒素检查法进行试验,将样品进行25~200倍稀释可以有效地消除其对鲎试验的干扰.结果与结论:用动态浊度法定量测定血栓通注射液中的细菌内毒素是可行的.

  19. Comparison of result judgment algorithm of test for interfering factors in the bacterial endotoxins test among Chinese, Japanese,European, American, and Indian pharmacopeias

    Institute of Scientific and Technical Information of China (English)

    Pei Yusheng; Cai Tong; Gao Hua; Tan Dejiang; Zhang Yuchen; Zhang Guolai

    2014-01-01

    Background 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).Methods 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.Results 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.Conclusions 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.

  20. SAGM红细胞保存液细菌内毒素检查法可行性研究%Study on the Feasibility of Bacterial Endotoxin Test for SAGM Erythrocyte Preserve Fluid

    Institute of Scientific and Technical Information of China (English)

    刘元书

    2001-01-01

    OBJECTIVE: To study the feasibility of bacterial endotoxin testfor SAGM erythrocyte preserve fluid as a substitute for pyrogen test.METHODS: Interference test was carried on endotoxin detection of SAGM erythrocyte preserve fluid with test agent of limulus(TAL).RESULTS: There was no interference action of twofold diluted SAGM fluid on limulus test.CONCLUSION: It is feasible to use 0.25EU/ml TAL in bacterial endotoxin test.%目的:研究SAGM红细胞保存液细菌内毒素检查法的可行性,以替代热原检查法。方法:按《中国药典》(二部)2000年版规定,用不同厂家生产的鲎试剂(TAL)对SAGM红细胞保存液进行细菌内毒素检查法干扰试验。结果:SAGM红细胞保存液经2倍稀释后对鲎试剂反应无干扰作用。结论:选用灵敏度为0.25EU/ml的鲎试剂进行细菌内毒素检查是可行的。

  1. pH值影响细菌内毒素测定的实验室研究%Effects of various pH in testing bacterial endotoxins

    Institute of Scientific and Technical Information of China (English)

    高国政; 颜锦

    1998-01-01

    OBJECTIVE: To study the effect of samples having different pH values in the bacterial endotoxins test (BET). METHODS: The BET was conducted as an assay of the concentration of endotoxins by using kinetic-turbidimetric technique. Samples having different pH values but containing same concentration reference standard endotoxin (RSE) were tested by TAL/LAL reagents. The average result on the standard curve was calculated and the mean recovery observed. RESULTS: The reagents had three highly sensitivities in pH 5.20, pH 7.83/pH 6.23 and pH 10.55. After adjustment to pH 6 to 8, the mean recovery was 76.5%~115.9% or 85.8%~99.8%. When the sample pH value was less than 3 or more than 12, the test was unsuitable for the inhibition test, and the mean recovery was less than 56.8%. CONCLUSION: It is necessary to adjust the pH of the solution to be tested in the BET, adjust the sample to pH (6.5~7.5) when using TAL reagents, and adjust to pH (6.2±0.1) or pH(6.7~7.8) when using LAL reagents.%目的:研究样品pH对细菌内毒素测定的影响。方法:应用动态浊度法定量测定样品中细菌内毒素的含量;由细菌内毒素参考标准品(RSE)制成两种浓度的稀释液,在14个pH量级上,与国内外两种鲎试剂(TAL,LAL)反应;观察每组平均回收率。结果:pH梯度上,两种鲎试剂回收曲线都呈现三个反应高峰,分别为pH 5.20,pH 7.83/pH 6.23和pH 10.55。样品pH在6~8间实验结果稳定,4λ阳性对照的平均回收率为76.5%~115.9%和85.8%~99.8%;当样品pH值≤3或≥12时,细菌内毒素试验受到抑制,平均回收率≤56.8%。结论:细菌内毒素试验中应该调节检品pH;使用TAL时,检品pH应预调在(6.5~7.5)为好;使用LAL时,检品pH应预调在(6.2±0.1)或(6.7~7.8)为好。

  2. Detection of Bacterial Endotoxins in Propacetamol Hydrochloride for Injection by Tachypleus Amebocyte Lysate Test%鲎试剂法检测注射用盐酸丙帕他莫中细菌内毒素

    Institute of Scientific and Technical Information of China (English)

    罗丽萍; 郭成希; 黄砚青

    2011-01-01

    Objective: To establish a method for detection of bacterial endotoxins in propacetainol hydrochloride for injection. Method: The experiments were carried out according to the bacterial endotoxins test specified in appendix XI E,volume two,Chinese pharmacopoeia 2010 edition. Result: Samples with the concentration of 5.0 mg·ml-1 had inhibitive effect on the bacterial endntoxins test,however the interference was eliminated when the samples were diluted to the concentration of 2. 5 mg·ml-1 ,detected hy tarhypleus ameboeyte lysates with the sensitivity of 0. 25 EU·ml-1. Conclusion: Tachypleus amebocyte lysate test is feasible for detecting bacterial endotoxins in propacetamol hydrochloride for injection.%目的:考察鲎试剂方法检测注射用盐酸丙帕他莫中细菌内毒素的可行性.方法:参照《中国药典》2010年版(二部)附录ⅪE细菌内毒素检查法对注射用盐酸丙帕他莫进行细菌内毒素检测.结果:质量浓度为5.0 mg·ml-1的样品溶液对凝集反应有抑制作用,质量浓度稀释至2.5 mg·ml-1时,用灵敏度λ为0.25 EU· ml-的鲎试剂进行试验,干扰作用消失.结论:注射用盐酸丙帕他莫可用鲎试剂方法进行细菌内毒素检测.

  3. Mediated effect of endotoxin and lead upon hepatic metabolism

    International Nuclear Information System (INIS)

    A test was made of the possibility that gram-negative bacterial cell wall lipopolysaccharides acted directly on key glucoregulatory enzymes in rat liver cytosol to cause the characteristic hypoglycemia of severe endotoxemia. Fasted male rats were sensitized to endotoxin by the simultaneous intravenous injection of lead acetate. The minimum systemic dosage of endotoxin necessary to perturb the normal pattern of hepatic glycolytic intermediates was determined by serial testing with diminishing dosages of endotoxin. The hepatocyte concentration of endotoxin was then calculated from this minimum dosage by use of literature data on the fraction of endotoxin delivered to liver cells after a systemic intravenous injection of radiochromium labeled lipopolysaccharides. Accepting a molecular weight of 118,000 daltons for the smallest endotoxin monomer capable of evoking a physiologic response, the molar amount of endotoxin present in 1 gram of hepatocytes was readily calculated. The concentration of glucoregulatory enzymes in parenchymal cells was then estimated from other literature sources. It was found that the amount of endotoxin in the hepatocytes was insufficient to combine directly with even 1 per cent of the quantity of a single key glucoregulatory enzyme in liver parenchyma. Since a one to one stoichiometric reaction between endotoxin and enzyme could not occur in the liver cytosol, a direct interaction mechanism between agonist and biocatalyst can be ruled out. It is concluded that bacterial endotoxin must act on hepatic glucoregulation by an indirect mechanism presumably based upon the release and operation of mediators

  4. Elevation in body temperature to fever range enhances and prolongs subsequent responsiveness of macrophages to endotoxin challenge.

    Directory of Open Access Journals (Sweden)

    Chen-Ting Lee

    Full Text Available Macrophages are often considered the sentries in innate immunity, sounding early immunological alarms, a function which speeds the response to infection. Compared to the large volume of studies on regulation of macrophage function by pathogens or cytokines, relatively little attention has been devoted to the role of physical parameters such as temperature. Given that temperature is elevated during fever, a long-recognized cardinal feature of inflammation, it is possible that macrophage function is responsive to thermal signals. To explore this idea, we used LPS to model an aseptic endotoxin-induced inflammatory response in BALB/c mice and found that raising mouse body temperature by mild external heat treatment significantly enhances subsequent LPS-induced release of TNF-α into the peritoneal fluid. It also reprograms macrophages, resulting in sustained subsequent responsiveness to LPS, i.e., this treatment reduces "endotoxin tolerance" in vitro and in vivo. At the molecular level, elevating body temperature of mice results in a increase in LPS-induced downstream signaling including enhanced phosphorylation of IKK and IκB, NF-κB nuclear translocation and binding to the TNF-α promoter in macrophages upon secondary stimulation. Mild heat treatment also induces expression of HSP70 and use of HSP70 inhibitors (KNK437 or Pifithrin-µ largely abrogates the ability of the thermal treatment to enhance TNF-α, suggesting that the induction of HSP70 is important for mediation of thermal effects on macrophage function. Collectively, these results support the idea that there has been integration between the evolution of body temperature regulation and macrophage function that could help to explain the known survival benefits of fever in organisms following infection.

  5. Effects of Endotoxin and Psychological Stress on Redox Physiology, Immunity and Feather Corticosterone in Greenfinches.

    Directory of Open Access Journals (Sweden)

    Richard Meitern

    Full Text Available Assessment of costs accompanying activation of immune system and related neuroendocrine pathways is essential for understanding the selective forces operating on these systems. Here we attempted to detect such costs in terms of disruption to redox balance and interference between different immune system components in captive wild-caught greenfinches (Carduelis chloris. Study birds were subjected to an endotoxin-induced inflammatory challenge and temporary exposure to a psychological stressor (an image of a predator in a 2*2 factorial experiment. Injection of bacterial endotoxin resulted in up-regulation of two markers of antioxidant protection - erythrocyte glutathione, and plasma oxygen radical absorbance (OXY. These findings suggest that inflammatory responses alter redox homeostasis. However, no effect on markers of oxidative damage to proteins or DNA in erythrocytes could be detected. We found no evidence that the endotoxin injection interfered with antibody production against Brucella abortus antigen or the intensity of chronic coccidiosis. The hypothesis of within-immune system trade-offs as a cost of immunity was thus not supported in our model system. We showed for the first time that administration of endotoxin can reduce the level of corticosterone deposited into feathers. This finding suggests a down-regulation of the corticosterone secretion cascade due to an endotoxin-induced immune response, a phenomenon that has not been reported previously. Exposure to the predator image did not affect any of the measured physiological parameters.

  6. Do calcium-mediated cellular signalling pathways, prostaglandin E2 (PGE2), estrogen or progesterone receptor antagonists, or bacterial endotoxins affect bovine placental function in vitro?

    Science.gov (United States)

    Weems, Y S; Randel, R D; Carstens, G E; Welsh, T H; Weems, C W

    2004-04-01

    .05). Concentrations of PGE2 in media at 4 and 8 h were lower (P or = 0.05). PGF2alpha was increased (P < or = 0.05) by RU-486 at 8h and no other treatment affected PGF2alpha at 4 or 8 h (P < or = 0.05). In conclusion, modulators of cellular calcium signalling pathways given alone do not affect bovine placental progesterone secretion at the days studied and progesterone receptor-mediated events appear to suppress placental progesterone, PGF2alpha, and PGE2 secretion in cattle. In addition, PGE2 does not appear to regulate bovine placental progesterone secretion when the corpus luteum is functional and bacterial endotoxin does not appear to affect bovine placental secretion of PGF2alpha or PGE2. PMID:15287156

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

    Science.gov (United States)

    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 ( 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 ( heifers following an endotoxin challenge and affected feedlot performance when incorporated in receiving diets in replacement of corn. Future studies will need to address strategies to increase DMI or explore levels of DCP less than 10% in the diet of newly received heifer calves. PMID:26641189

  8. Establishment of Bacterial Endotoxin Test of Iodine [131 I]Actuximab Injection%碘[131I]爱克妥昔单抗注射液细菌内毒素检查方法建立

    Institute of Scientific and Technical Information of China (English)

    孙祥敏; 张云; 张先; 杨薇薇; 蔡海燕

    2015-01-01

    为建立碘[131 I]爱克妥昔单抗注射液的细菌内毒素检查方法,采用《中国药典》二部细菌内毒素检查法进行实验。结果表明,将供试品稀释10倍、20倍、40倍进行细菌内毒素检查均存在干扰;对供试品稀释80倍进行检查或采用抗增液复溶鲎试剂,再将供试品稀释20倍进行检查,均无干扰。供试品的细菌内毒素检查可采用将供试品至少稀释80倍进行检验的方法;也可采用抗增液复溶鲎试剂,将供试品稀释至少20倍进行检查。结果表明,直接稀释法为产品检验的首选方法。%To establish a method for the bacterial endotoxin test of Iodine [1 3 1 I]Actux-imab Injection,The experiment was carried out according to the appendix ⅪE in Vol.Ⅱof the 2010 version of Chinese Pharmacopoeia.The results demonstrated that the prepa-ration interfered with the test at 10 times,20 times and 40 times dilution,and the inter-ference can be eliminated after the sample was diluted to 80 times or the sample was diluted to 20 times with the endotoxin-specific buffer dissolving the Tachypleus Amebo-cyte Lysate.Two methods can be used to test the bacterial endotoxin of Iodine [1 3 1 I] Actuximab Injection.One is the sample should be diluted to atleast 80 times,the other is the sample was diluted to 20 times with the endotoxin-specific buffer dissolving the Tachypleus Amebocyte Lysate.The dilution method is the preferred method for the quality control.

  9. 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: No

  10. 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 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.

  11. Parasitic infection protects wasp larvae against a bacterial challenge.

    Science.gov (United States)

    Manfredini, Fabio; Beani, Laura; Taormina, Mauro; Vannini, Laura

    2010-09-01

    Host antibacterial defense after Strepsiptera parasitization is a complex and rather unexplored topic. The way how these parasites interact with bacteria invading into the host insect during an infection is completely unknown. In the present study we demonstrate that larvae of the paper wasp Polistes dominulus are more efficient at eliminating bacteria when they are parasitized by the strepsipteran insect Xenos vesparum. We looked at the expression levels of the antimicrobial peptide defensin and we screened for the activity of other hemolymph components by using a zone of inhibition assay. Transcription of defensin is triggered by parasitization, but also by mechanical injury (aseptic injection). Inhibitory activity in vitro against the Gram positive bacterium Staphylococcus aureus is not influenced by the presence of the parasite in the wasp or by a previous immune challenge, suggesting a constitutive power of killing this bacterium by wasp hemolymph. Our results suggest either direct involvement of the parasite or that defensin and further immune components not investigated in this paper, for example other antimicrobial peptides, could play a role in fighting off bacterial infections in Polistes. PMID:20546915

  12. Methods of Endotoxin Detection.

    Science.gov (United States)

    Su, Wenqiong; Ding, Xianting

    2015-08-01

    Endotoxin, present in the outer membrane of all gram-negative bacteria, can pose serious risks to human health, from irreversible shock to death. Therefore, it is essential to develop sensitive, accurate, and rapid methods for its detection. The rabbit pyrogen test is the first standard technique for endotoxin detection and, nowadays, has been replaced by the Limulus Amoebocyte Lysate test, which is the most popular detection technique for endotoxin. With in-depth understanding of endotoxin, biosensors based on endotoxin-sensing components are promising alternatives to pursue in developing low-cost, easy-operation, and fast-response endotoxin detection techniques. This article summarizes the recent advances of endotoxin detection methods with a particular emphasis on optical and electrochemical biosensors based on various sensing elements ranging from nature biomolecules to artificial materials. As the research and technological revolution continues, the highly integrated and miniaturized commercial devices for sensitively and reliably detecting endotoxin will provide a wide range of applications in people's daily life. PMID:25720597

  13. Endotoxins in the prostatic secretions of chronic prostatitis patients

    Institute of Scientific and Technical Information of China (English)

    Yu-Ping Dai; Xiang-Zhou Sun; Ke-Li Zheng

    2005-01-01

    Aim: To evaluate the clinical significance of the quantitative determinations of endotoxins in the expressed prostatic secretions (EPS) of chronic prostatitis (CP) patients. Methods: The EPS of 45 patients with CP and 15 normal volunteers were obtained for microscopic examination, bacterial culture and endotoxin determination. The level of endotoxins was determined by the Limulus-amebocyte-lysate test with chromogenic substrate. Results: Patients (P>0.05), type Ⅱ/type Ⅲa vs. Normal controls P < 0.05)]. Conclusion: CP patients have elevated levels of endotoxins in the EPS, which suggests that inflammation is a feature of this disease. EPS endotoxin determination is not only helpful in diagnostic confirmation, but also in evaluating the response to treatment in CP patients.

  14. Effect of the dilution factor on {sup 18}FDG and Na{sup 18}F samples for bacterial endotoxin test using PTS (portable test system)

    Energy Technology Data Exchange (ETDEWEB)

    Silveira, Marina B.; Costa, Flavia M.; Ferreira, Soraya Z., E-mail: mbs@cdtn.b [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN-MG), Belo Horizonte, MG (Brazil). Unidade de Pesquisa e Producao de Radiofarmacos

    2011-07-01

    {sup 18}FDG and Na{sup 18}F are radiopharmaceuticals produced as sterile solutions suitable for intravenous administration, which must contain no more than 175 EV/V. The most commonly used approach to detect endotoxins is the gelclot technique that requires 60 minutes for results. For radiopharmaceuticals containing short-life radionuclides, such as {sup 18}F, there is an increasing interest for faster quality control methods. FDA licensed the Endosafe, PTS, a kinetic chromogenic endotoxin detection system that takes about 15 minutes for results. As other techniques, PTS test is susceptible to interferences which can be solved by product dilution. The aim of this study was to establish the best dilution of {sup 18}FDG and Na{sup 18}F for PTS analysis. Two different dilution factors for {sup 18}FDG and 1:10 for Na{sup 18}F were essayed: 1:10 and 1:100. {sup 18}FDG and Na{sup 18} solutions were prepared by the addition of LAL reagent water. Considering the assay acceptance criteria, the best dilution factor was 1:100 for {sup 18}FDG and 1:10 for Na{sup 18}F. The recovery of the product positive control was 98-12% for {sup 18}FDG 1:100 and 104-120% for Na{sup 18}F 1:10, which were, in both cases, within the specification (50-200%) and very close to 100%. Results obtained with these dilution studies were important to establish the most appropriate and non-interfering dilution factor for {sup 18}FDG and Na{sup 18}F routine endotoxin test. (author)

  15. Effect of the dilution factor on 18FDG and Na18F samples for bacterial endotoxin test using PTS (portable test system)

    International Nuclear Information System (INIS)

    18FDG and Na18F are radiopharmaceuticals produced as sterile solutions suitable for intravenous administration, which must contain no more than 175 EV/V. The most commonly used approach to detect endotoxins is the gelclot technique that requires 60 minutes for results. For radiopharmaceuticals containing short-life radionuclides, such as 18F, there is an increasing interest for faster quality control methods. FDA licensed the Endosafe, PTS, a kinetic chromogenic endotoxin detection system that takes about 15 minutes for results. As other techniques, PTS test is susceptible to interferences which can be solved by product dilution. The aim of this study was to establish the best dilution of 18FDG and Na18F for PTS analysis. Two different dilution factors for 18FDG and 1:10 for Na18F were essayed: 1:10 and 1:100. 18FDG and Na18 solutions were prepared by the addition of LAL reagent water. Considering the assay acceptance criteria, the best dilution factor was 1:100 for 18FDG and 1:10 for Na18F. The recovery of the product positive control was 98-12% for 18FDG 1:100 and 104-120% for Na18F 1:10, which were, in both cases, within the specification (50-200%) and very close to 100%. Results obtained with these dilution studies were important to establish the most appropriate and non-interfering dilution factor for 18FDG and Na18F routine endotoxin test. (author)

  16. Extraction of Bacterial RNA from Soil: Challenges and Solutions

    OpenAIRE

    Wang, Yong; Hayatsu, Masahito; Fujii, Takeshi

    2012-01-01

    Detection of bacterial gene expression in soil emerged in the early 1990s and provided information on bacterial responses in their original soil environments. As a key procedure in the detection, extraction of bacterial RNA from soil has attracted much interest, and many methods of soil RNA extraction have been reported in the past 20 years. In addition to various RT-PCR-based technologies, new technologies for gene expression analysis, such as microarrays and high-throughput sequencing techn...

  17. Application of bacterial endotoxin test to the safety evaluation of clinical use of hydrocolloid dressing%细菌内毒素检查法在水胶体敷料临床使用安全性评价中的应用

    Institute of Scientific and Technical Information of China (English)

    许雷鸣; 顾倩; 武谷; 堵伟锋; 李翔; 林园园

    2014-01-01

    目的:建立水胶体敷料的细菌内毒素检查方法,为评价临床使用的安全性提供依据。方法按水胶体敷料表面积每1 cm2加入4 mL细菌内毒素检查用水,37℃浸提2 h,用2个不同厂家的鲎试剂进行干扰试验研究并采用凝胶半定量试验测定产品中的细菌内毒素含量。结果根据临床实际应用情况,确定水胶体敷料的内毒素限值L=0.5 EU·cm-2;在本实验条件下,可用灵敏度为0.125 EU·mL-1及0.125 EU·mL-1以上的鲎试剂检测水胶体敷料中的细菌内毒素。结论本试验建立的细菌内毒素检查方法可用于水胶体敷料的细菌内毒素检查,保障其临床使用的安全性。%Objective To establish a bacterial endotoxin test method for hydrocolloid dressing and to provide the experimental basis forthe safety evaluation of clinical use.Methods Each 1 cm2 total surface area of sample was added with 4 mL water for bacterial endo-toxin test and was extracted by heating at 37℃ for 2 hours.Tachypleus amebocyte lysate from two manufacturers were used for interfer-ence test and semi-quantitative test was used in the endotoxin assay of hydrocolloid dressing.Results According to the dose of clinicalapplication,the most suitable limit of bacterial endotoxin for hydrocolloid dressing was 0.5 EU·cm -2 .Under this experimental condi-tion,it was effective that bacterial endotoxin was tested by tachypleus amebocyte lysate with the sensitivity of 0.125 EU·mL -1 or high-er sensitivity.Conclusions This method can be used in bacterial endotoxin test of hydrocolloid dressing and can be applied to securethe safety of clinical use.

  18. Transfer of immunoglobulins through the mammary endothelium and epithelium and in the local lymph node of cows during the initial response after intramammary challenge with E. coli endotoxin

    Directory of Open Access Journals (Sweden)

    Lun Shichun

    2008-07-01

    Full Text Available Abstract Background The first hours after antigen stimulation, interactions occur influencing the outcome of the immunological reaction. Immunoglobulins originate in blood and/or are locally synthesized. The transfer of Ig isotypes (Igs in the udder has been studied previously but without the possibility to distinguish between the endothelium and the epithelium. The purpose of this study was to map the Ig transfer through each barrier, separately, and Ig transfer in the local lymph nodes of the bovine udder during the initial innate immune response. Methods The content of IgG1, IgG2, IgM, IgA and albumin (BSA was examined in peripheral/afferent mammary lymph and lymph leaving the supramammary lymph nodes, and in blood and milk before (0 h and during 4 hours after intramammary challenge with Esherichia coli endotoxin in 5 cows. Results Igs increased most rapidly in afferent lymph resulting in higher concentrations than in efferent lymph at postinfusion hour (PIH 2, contrary to before challenge. Ig concentrations in milk were lower than in lymph; except for IgA at 0 h; and they increased more slowly. Afferent lymph:serum and efferent lymph:serum concentration ratios (CR of Igs were similar to those of BSA but slightly lower. Milk:afferent lymph (M:A CRs of each Ig, except for IgG2, showed strikingly different pattern than those of BSA. The M:A CR of IgG1, IgM and IgA were higher than that of BSA before challenge and the CR of IgA and IgG1 remained higher also thereafter. At PIH 2 there was a drop in Ig CRs, except for IgG2, in contrast to the BSA CR which gradually increased. The M:A CR of IgM and Ig A decreased from 0 h to PIH 4, in spite of increasing permeability. Conclusion The transfer of Igs through the endothelium appeared to be merely a result of diffusion although their large molecular size may hamper the diffusion. The transfer through the epithelium and the Ig concentrations in milk seemed more influenced by selective mechanisms and

  19. Comparative effects of dietary nucleoside-nucleotide mixture and its components on endotoxin induced bacterial translocation and small intestinal injury in protein deficient mice.

    OpenAIRE

    Adjei, A A; Yamauchi, K.; Chan, Y. C.; Konishi, M; Yamamoto, S.

    1996-01-01

    BACKGROUND--Nucleoside-nucleotide mixture has been shown to improve gut morphology and reduce the incidence of bacterial translocation in protein deficient mice. AIMS--To compare the reparative effect of nucleoside-nucleotide mixture and their individual components on maintenance of gut integrity and bacterial translocation based on their differential metabolism and utilisation. METHODS--ICR (CD-1) mice were randomised into eight groups of 10 animals each and fed 20% casein diet (control), pr...

  20. Biosensor of endotoxin and sepsis

    Science.gov (United States)

    Shao, Yang; Wang, Xiang; Wu, Xi; Gao, Wei; He, Qing-hua; Cai, Shaoxi

    2001-09-01

    To investigate the relation between biosensor of endotoxin and endotoxin of plasma in sepsis. Method: biosensor of endotoxin was designed with technology of quartz crystal microbalance bioaffinity sensor ligand of endotoxin were immobilized by protein A conjugate. When a sample soliton of plasma containing endotoxin 0.01, 0.03, 0.06, 0.1, 0.5, 1.0Eu, treated with perchloric acid and injected into slot of quartz crystal surface respectively, the ligand was released from the surface of quartz crystal to form a more stable complex with endotoxin in solution. The endotoxin concentration corresponded to the weight change on the crystal surface, and caused change of frequency that occurred when desorbed. The result was biosensor of endotoxin might detect endotoxin of plasma in sepsis, measurements range between 0.05Eu and 0.5Eu in the stop flow mode, measurement range between 0.1Eu and 1Eu in the flow mode. The sensor of endotoxin could detect the endotoxin of plasm rapidly, and use for detection sepsis in clinically.

  1. Hacking into bacterial biofilms: a new therapeutic challenge.

    Science.gov (United States)

    Bordi, Christophe; de Bentzmann, Sophie

    2011-06-13

    Microbiologists have extensively worked during the past decade on a particular phase of the bacterial cell cycle known as biofilm, in which single-celled individuals gather together to form a sedentary but dynamic community within a complex structure, displaying spatial and functional heterogeneity. In response to the perception of environmental signals by sensing systems, appropriate responses are triggered, leading to biofilm formation. This process involves various molecular systems that enable bacteria to identify appropriate surfaces on which to anchor themselves, to stick to those surfaces and to each other, to construct multicellular communities several hundreds of micrometers thick, and to detach from the community. The biofilm microbial community is a unique, highly competitive, and crowded environment facilitating microevolutionary processes and horizontal gene transfer between distantly related microorganisms. It is governed by social rules, based on the production and use of "public" goods, with actors and recipients. Biofilms constitute a unique shield against external aggressions, including drug treatment and immune reactions. Biofilm-associated infections in humans have therefore generated major problems for the diagnosis and treatment of diseases. Improvements in our understanding of biofilms have led to innovative research designed to interfere with this process.

  2. 乙型脑炎减毒活疫苗细菌内毒素检查方法及质量标准的建立%Establishment of the inspection methods and quality criteria for bacterial endotoxin of live-attenuated vaccine of encephalitis B virus

    Institute of Scientific and Technical Information of China (English)

    李红芳; 陈继军; 王建军; 王伟; 杨博涵; 李守丽; 邹勇

    2009-01-01

    目的 建立乙型脑炎减毒活疫苗细菌内毒素检测的方法及质量标准.方法 参照(三部)中细菌内毒素检查法检测,并与家兔热原实验结果进行对比.结果 使用凝胶限量法检测乙型脑炎减毒活疫苗细菌内毒素,限值为40 EU/mL,即将样品稀释至160倍,在该稀释度下样品对试验没有干扰.结论 该方法简便、灵敏、结果可靠,可用于乙型脑炎减毒活疫凿中间产品的细菌内毒素检查.%Objective To establish the inspection methods and quality criteria for bacterial en- dotoxin of live-attenuated vaccine of encephalitis B virus. Methods The inspecting method for bacterial endotoxin from Pharmacopoeia of the People's Republic of China (3 ed) was employed and the results were compared with the results of rabbit pyrogen test. Results Bacterial endo- toxin of live-attenuated vaccine of encephalitis B virus was detected by gel-clot limit test,the detection limit was 40 EU/mL,namely,160-fold dilution of the sample. In this dilution,no inter- ference of samples to the endotoxin test was observed. Conclusion The method is convenient, sensitive and reliable and may be used for bacterial endotoxin inspection of intermediate products of live-attenuated vaccine of encephalitis B virus.

  3. 凝胶法和动态浊度法在b型流感嗜血杆菌结合疫苗细菌内毒素控制中的应用比较%Comparison of Gel-clot Test and Kinetic Turbidinetric Method Application in the Control of Bacterial Endotoxin in Hib

    Institute of Scientific and Technical Information of China (English)

    马平; 马国荣; 吴菊寿; 齐娟; 杜送田

    2013-01-01

    Objective To detect the content of bacterial endotoxin in the intermediate, final bulk and finishied products of haemophilus type B conjugate vaccine with Kinetic turbidinetric method and gel clot test for the control of pyrogen and improving the quality of the products .Meanwhile compare with two methods. Method According to the Methods of Detection of Bacterial Endotoxin in the AppendixⅫE of The Pharmacopoeia of the People’s Republic of China of the year 2010 edition.Results The detection limit for acterial endotoxin of the intermediate,final bulk and finishied products of Hib defined 25EU/µg、5 EU/µg、25 EU/dose,in accordance with the provisions. Conclusion The results of the tests indicate that application of kinetic turbidinetric method and gel clot test for detection of Haemophilus influenza type B conjugates vaccine intermediates,final bulk and finished products in accordance with the results of the content of bacterial endotoxin,quantitative Kinetic turbidinetric method is a good guidance for endotoxin content of internal control index.%目的:在b型流感嗜血杆菌结合疫苗生产过程中采用凝胶法和动态浊度法检测中间品、原液和成品中细菌内毒素含量,以此控制热原,提高制品质量,并对两种方法进行比较。方法采用《中华人民共和国药典》(2010年版)三部附录ⅫE细菌内毒素检查法中的凝胶法和动态浊度法。结果 Hib中间品、原液及成品经凝胶法和动态浊度法测定,内毒素含量均<25EU/µg、5EU/µg、25EU/剂,符合规定。结论各项检定结果表明:应用动态浊度法和凝胶法检测b型流感嗜血杆菌结合疫苗中间品、原液和成品细菌内毒素含量结果相符,动态浊度法定量检测对于内毒素含量内控指标有很好的指导意义。

  4. Estimativa da incerteza em ensaio de detecção de endotoxina bacteriana pelo método de gelificação Estimation of uncertainty in the detection of bacterial endotoxin by gel-clot method

    Directory of Open Access Journals (Sweden)

    Felipe Rebello Lourenço

    2005-12-01

    Full Text Available Desde a publicação da ISO 17025:1999, o interesse em métodos para estimativa da incerteza em ensaios qualitativos, do tipo "passa/não passa", têm ganho grande importância. Uma forma de estimar e informar a incerteza deste tipo de ensaio é o uso das probabilidades de respostas-falsas, particularmente falsos-positivos e falsos-negativos, determinados a partir do teorema de Bayes. O objetivo deste artigo é estabelecer um método para a estimativa de incerteza em ensaios de detecção de endotoxina bacteriana pelo método in vitro Limulus Amebocyte Lysate (LAL. Considerando a confirmação da sensibilidade do LAL e a validação do teste, a probabilidade de uma resposta falsa corresponde à soma da probabilidade dos resultados falso-negativos e falso-positivos. A partir dos resultados obtidos foi verificado que a etapa da confirmação da sensibilidade do LAL contribui para a incerteza de forma mais significativa (67,6% que a etapa de validação do teste (32,4%. Através de um procedimento simples, descrito neste artigo, e de dados obtidos a partir da confirmação da sensibilidade do LAL e validação do teste para um produto em questão é possível obter uma estimativa de incerteza razoável para o ensaio de detecção de endotoxinas bacterianas pelo método de gelificação.Since the publication of ISO 17025:1999, the interest in methods for estimation of the uncertainty in qualitative analysis, such as 'pass/fail', have became more important. The usual form of estimating and informing the uncertainty in this kind of analysis is the use of false-response rates, particularly false-positive and false-negative, determinated from Bayes theorem. The aim of this paper is establish a method for estimation of the uncertainty in the detection of bacterial endotoxins by in vitro Limulus Amebocyte Lysate (LAL test. Considering the confirmation of LAL sensitivity and the validation of the test, the probability of a false-response corresponds to the

  5. [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.

  6. 动态浊度法检测乳酸左氧氟沙星氯化钠注射液中的细菌内毒素%Determination of Bacterial Endotoxin for Levofloxacin Lactate/Sodium Chloride Injection by Kinetic Turbidimetric Limulus Test

    Institute of Scientific and Technical Information of China (English)

    李荣; 吴玲

    2012-01-01

    OBJECTIVE: To apply kinetic turbidimetric limulus test for quantitative determination of bacterial endotoxin for Le-vofloxacin lactate/sodium chloride injection, compared with the gel-clot. METHODS: According to bacterial endotoxin test included in Chinese Pharmacopoeia (2010 Edition) , interference test was conducted to determine maximum diluent fold. RESULTS: When the sample was 8-fold diluted, the recovery rate of kinetic turbidimetric limulus was high relatively (about 100%) without interference. The method and gel-clot were consistent and compliance, and both of them were up to the standard. CONCLUSION: Kinetic turbidimetric limulus test can effectively measure bacterial endotoxin for Levofloxacin lactate/sodium chloride injection.%目的:应用动态浊度法定量测定乳酸左氧氟沙星氯化钠注射液中的细菌内毒素,并与凝胶法比较.方法:按《中国药典》2010年版二部收载的细菌内毒素检查方法及其指导原则进行干扰试验,确定样品最大稀释倍数.结果:当样品作8倍稀释时用动态浊度法的回收率值较高(100%左右),未见干扰作用;与凝胶法检测结果一致,均符合规定.结论:动态浊度法可以有效测定乳酸左氧氟沙星氯化钠注射液中的细菌内毒素含量.

  7. Rapid diagnosis of gram negative pneumonia by assay of endotoxin in bronchoalveolar lavage fluid.

    Science.gov (United States)

    Pugin, J; Auckenthaler, R; Delaspre, O; van Gessel, E; Suter, P M

    1992-01-01

    BACKGROUND: Diagnosis of ventilator associated pneumonia can be made by quantitative cultures of bronchoalveolar lavage fluid or of protected specimen brushings, though cultures require 24-48 hours to provide results. In 80% of cases aerobic Gram negative bacteria are the cause. METHODS: A rapid diagnostic method of assessing the endotoxin content of lavage fluid by Limulus assay is described. Forty samples of lavage fluid were obtained from patients with multiple trauma requiring mechanical ventilation for a prolonged period. Pneumonia was diagnosed on the basis of clinical, radiological, and bacteriological findings, including quantitative cultures of lavage fluid. RESULTS: A relation was observed between the concentration of endotoxin in lavage fluid and the quantity of Gram negative bacteria. The median endotoxin content of lavage fluid in Gram negative bacterial pneumonia was 15 endotoxin units (EU)/ml; the range observed in individual patients was 6 to > 150 EU/ml. In patients with pneumonia due to Gram positive cocci and in non-infected patients the median endotoxin level was 0.17 (range < or = 0.06 to 2) EU/ml. An endotoxin level greater than or equal to 6 EU/ml distinguished patients with Gram negative bacterial pneumonia from colonised patients and from those with pneumonia due to Gram positive cocci. CONCLUSION: The measurement of endotoxin in lavage fluid is a rapid (less than two hours) and accurate diagnostic method. It should allow specific and early treatment of Gram negative bacterial pneumonia. PMID:1412100

  8. ENDOTOXIN ADSORPTION IN SEPSIS IN PATIENTS AFTER RENAL TRANSPLANTATION AND GRAFT FUNCTION

    Directory of Open Access Journals (Sweden)

    A. V. Vatazin

    2012-01-01

    Full Text Available Objective: to evaluate the influence of endotoxin adsorption on graft function and mortality. Methods: we analy- zed the results of treatment of 90 patients with bacterial complications after renal transplantation and compared them with retrospective group of patients, who didn’t undergo endotoxin adsorption. Results: the use of endo- toxin adsorption led to significant improvement of graft function and intraorganic microcirculation in compa- rison with retrospective group, treated without endotoxin adsorption. Survival was higher in the intervention group. However, endotoxin adsorption did not affect patient survival when multiple organ failure developed. Conclusion: when infectious complications developed, endotoxin adsorption is able to improve the graft function and recipient survival significantly. The earliest application of this procedure is recommended. 

  9. 动态浊度法定量测定黄芪注射液中细菌内毒素的含量%Detection of Bacterial Endotoxin in Huangqi Injection by the Method of Kinetic Turbidimetric Limulus Test

    Institute of Scientific and Technical Information of China (English)

    卢文斌

    2012-01-01

    OBJECTIVE To detect the quantity of endotoxin in huangqi injection by the method of kinetic turbidimetric limulus test. METHODS The kinetic turbidimetric limulus test was applied to detect the quantity of endotoxin in several samples. The standard endotoxin working curve was set up and the interference test was studied to range the effective concentrations of sample. RESULTS Dilution ratio of Huangqi injection at 1 : 5 show no inference in the test. The endotoxin recovery rate was more than 50% and less than 200%. The endotoxin level in samples could be quantified exactly, and the results were accorded with the results obtained by gel-clot technique assay. CONCLUSION The kinetic turbidimetric limulus test can be used to quantitate the endotoxin level in huangqi injection.%目的 应用动态浊度法尝试验定量测定黄芪注射液中的细菌内毒素含量.方法 通过干扰试验确定样品检测浓度,并建立标准曲线对不同的样品进行细菌内毒素的定量测定.结果 样品用BET水制成1∶5稀释液,对鲎试验检查无干扰,细菌内毒素回收率在50% ~ 200%之间.样品中的内毒素含量可定量测定,检测结果与凝胶法检测的结果有一定的相关性.结论 动态浊度法可用于黄芪注射液中细菌内毒素定量检测.

  10. The Effect of Residual Endotoxin Contamination on the Neuroinflammatory Response to Sterilized Intracortical Microelectrodes.

    Science.gov (United States)

    Ravikumar, Madhumitha; Hageman, Daniel J; Tomaszewski, William H; Chandra, Gabriella M; Skousen, John L; Capadona, Jeffrey R

    2014-05-01

    A major limitation to the use of microelectrode technologies in both research and clinical applications is our inability to consistently record high quality neural signals. There is increasing evidence that recording instability is linked, in part, to neuroinflammation. A number of factors including extravasated blood products and macrophage released soluble factors are believed to mediate neuroinflammation and the resulting recording instability. However, the roles of other inflammatory stimuli, such as residual endotoxin contamination, are poorly understood. Therefore, to determine the effect of endotoxin contamination we examined the brain tissue response of C57/BL6 mice to non-functional microelectrodes with a range of endotoxin levels. Endotoxin contamination on the sterilized microelectrodes was measured using a limulus amebocyte lysate test following FDA guidelines. Microelectrodes sterilized by autoclave, dry heat, or ethylene oxide gas, resulted in variable levels of residual endotoxins of 0.55 EU/mL, 0.22 EU/mL, and 0.11 EU/mL, respectively. Histological evaluation at two weeks showed a direct correlation between microglia/macrophage activation and endotoxin levels. Interestingly, astrogliosis, neuronal loss, and blood brain barrier dysfunction demonstrated a threshold-dependent response to bacterial endotoxins. However, at sixteen weeks, no histological differences were detected, regardless of initial endotoxin levels. Therefore, our results demonstrate that endotoxin contamination, within the range examined, contributes to initial but not chronic microelectrode associated neuroinflammation. Our results suggest that minimizing residual endotoxins may impact early recording quality. To this end, endotoxins should be considered as a potent stimulant to the neuroinflammatory response to implanted intracortical microelectrodes.

  11. 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...

  12. Microfiltration, Nano-filtration and Reverse Osmosis for the Removal of Toxins (LPS Endotoxins) from Wastewater

    OpenAIRE

    Mokhtar, Guizani; Naoyuki, Funamizu

    2012-01-01

    Lipopolysaccharide (LPS) endotoxin, a bacterial byproduct abundantly present in wastewater, is more and more representing a major concern in wastewater treatment sector for the potential health risk it represents. It is, therefore, more urgent than before to protect consumers from contaminating their fresh potable water reserves with LPS endotoxin through aquifer replenishment using reclaimed wastewater or by supplying reclaimed wastewater as potable water. Membrane treatment is an alternativ...

  13. Detection of free endotoxin in cerebrospinal fluid by the Limulus lysate test.

    OpenAIRE

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

    1984-01-01

    We used a rabbit model of Escherichia coli meningitis to study the basis for positive Limulus lysate tests in infected cerebrospinal fluid. The results indicated that positive Limulus tests are due to endotoxins in cerebrospinal fluid and not to leukocyte proteases or other possible activators of the Limulus clotting system. The results also suggest that bacteria-free endotoxin may be present in localized gram-negative bacterial infections.

  14. Comparative Resistance of AH26 and a New Sealer Prototype to a Bacterial Challenge

    Directory of Open Access Journals (Sweden)

    Derek Duggan

    2012-01-01

    Full Text Available Objective. This study compared the leakage resistance of a New Sealer Prototype (NSP with a traditional sealer (AH 26 in Resilon-filled roots subjected to a bacterial challenge. Study Design. 41 roots were instrumented to ISO size 50 apically. Group 1 (=20 contained Resilon and AH 26 sealer and roots in group 2 (=21 contained Resilon and NSP. Roots were embedded in a dual-chamber model with the upper chamber containing Streptococcus mutans inoculum. Evidence of bacterial penetration was observed for 1 month. Fisher's Test was used to analyze the data. Results. 8 of 20 roots (40% in the AH 26 group demonstrated leakage whereas 3 of 21 roots (14% in the NSP group leaked. The difference in leakage rates was not statistically significant (=0.053. Conclusion. The traditional sealer (AH 26 demonstrated increased leakage rates compared to the New Sealer Prototype (NSP, but the difference did not reach statistical significance in this study.

  15. Bacterial Glycosyltransferases: Challenges and opportunities of a highly diverse enzyme class toward tailoring natural products

    Directory of Open Access Journals (Sweden)

    Jochen eSchmid

    2016-02-01

    Full Text Available The enzyme subclass of glycosyltransferases (EC 2.4 currently comprises 97 families as specified by CAZy classification. One of their important roles is in the biosynthesis of disaccharides, oligosaccharides and polysaccharides by catalyzing the transfer of sugar moieties from activated donor molecules to other sugar molecules. In addition glycosyltransferases also catalyze the transfer of sugar moieties onto aglycons, which is of great relevance for the synthesis of many high value natural products. Bacterial glycosyltransferases show a higher sequence similarity in comparison to mammalian ones. Even when most glycosyltransferases are poorly explored, state of the art technologies, such as protein engineering, domain swapping or computational analysis strongly enhance our understanding and utilization of these very promising classes of proteins. This perspective article will focus on bacterial glycosyltransferases, especially on classification, screening and engineering strategies to alter substrate specificity. The future development in these fields as well as obstacles and challenges will be highlighted and discussed.

  16. The mucosal expression signatures of g-type lysozyme in turbot (Scophthalmus maximus) following bacterial challenge.

    Science.gov (United States)

    Gao, Chengbin; Fu, Qiang; Zhou, Shun; Song, Lin; Ren, Yichao; Dong, Xiaoyu; Su, Baofeng; Li, Chao

    2016-07-01

    The mucosal surfaces constitute the first line of host defense against infection, and also serve as the dynamic interfaces that simultaneously mediate a diverse array of critical physiological processes, while in constantly contact with a wide range of pathogens. The lysozymes are considered as key components for innate immune response to pathogen infection with their strong antibacterial activities. But their activities in mucosal immune responses were always overlooked, especially for g-type lysozymes, whose expression patterns in mucosal tissues following bacterial challenge are still limited. Towards to this end, here, we characterized the g-type lysozymes, Lyg1 and Lyg2 in turbot, and determined their expression patterns in mucosal barriers following different bacterial infection. The phylogenetic analysis revealed the turbot g-type lysozyme genes showed the closest relationship to Cynoglossus semilaevis. The two lysozyme genes showed different expression patterns following challenge. Lyg2 was significantly up-regulated in mucosal tissues following Vibrio anguillarum and Streptococcus iniae challenge, while Lyg1 showed a general trend of down-regulation. The significant mucosal expression signatures of g-type lysozyme genes indicated their key roles to prevent pathogen attachment and entry in the first line of host defense system. Further functional studies should be carried out to better characterize the availability of utilization of g-type lysozyme to increase the disease resistance in the mucosal surfaces and facilitate the disease resistant breeding selection. PMID:27189917

  17. Fiber-optic Fourier transform infrared (FO-FTIR) spectroscopy for detecting endotoxin contamination in ophthalmic viscosurgical devices (OVDS) (Conference Presentation)

    Science.gov (United States)

    Hassan, Moinuddin; Ilev, Ilko

    2016-03-01

    Ophthalmic Viscosurgical Devices (OVDs) in clinical setting are a major health risk factor for potential endotoxin contamination in the eye, due to their extensive applications in cataract surgery for space creation, stabilization and protection of intraocular tissue and intraocular lens (IOL) during implantation. Endotoxin contamination of OVDs is implicated in toxic anterior syndrome (TASS), a severe complication of cataract surgery that leads to intraocular damage and even blindness. Current standard methods for endotoxin contamination detection utilize rabbit assay or Limulus amoebocyte lysate (LAL) assays. These endotoxin detection strategies are extremely difficult for gel-like type devices such as OVDs. To overcome the endotoxin detection limitations in OVDs, we have developed an alternative optical detection methodology for label-free and real-time sensing of bacterial endotoxin in OVDs, based on fiber-optic Fourier transform infrared (FO-FTIR) transmission spectrometry in the mid-IR spectral range from 2.5 micron to 12 micron. Endotoxin contaminated OVD test samples were prepared by serial dilutions of endotoxins on OVDs. The major results of this study revealed two salient spectral peak shifts (in the regions 2925 to 2890 cm^-1 and 1125 to 1100 cm^-1), which are associated with endotoxin in OVDs. In addition, FO-FTIR experimental results processed using a multivariate analysis confirmed the observed specific peak shifts associated with endotoxin contamination in OVDs. Thus, employing the FO-FTIR sensing methodology integrated with a multivariate analysis could potentially be used as an alternative endotoxin detection technique in OVD.

  18. New generation ceramic membranes have the potential of removing endotoxins from dialysis water and dialysate.

    Science.gov (United States)

    Czermak, P; Ebrahimi, M; Catapano, G

    2005-07-01

    Poor water properties, use of concentrated bicarbonate, and biofilm growth in pipes and storage tanks often cause dialysis water and dialysate contamination with bacteria and endotoxins. High-flux dialysis with bicarbonate may favor endotoxin transfer from the dialysate into the blood exposing patients to serious short-and long-term side effects. Ultrafiltration across hydrophobic synthetic membranes effectively removes endotoxins from dialysis water by combined filtration and adsorption. However, repeated sterilization worsens the membrane separation properties,and limits their use. Ceramic membranes are generally more resistant to harsh operating conditions than polymeric membranes, and may represent an alternative for endotoxin removal. Previously, we proved that the ceramic membranes commercially available at that time were not retentive enough to ensure production of endotoxin-free dialysis water. In this paper, we investigated the endotoxin removal capacity of new generation commercial ceramic membranes with nominal molecular weight cut-off down to 1,000. In dead-end filtration, all investigated membranes produced water meeting, the European standards, or close to,when challenged with low endotoxin concentrations, but only one membrane type succeeded at high endotoxin concentrations. In cross-flow filtration, none produced water meeting the European standard. Moreover, sterilization and rinsing procedures altered the separation properties of two out of three membrane types. PMID:16049903

  19. 动态浊度法鲎试验用于检测人凝血因子Ⅷ制品中细菌内毒素的研究%Determination of bacterial endotoxin in human coagulation factor Ⅷ by kinetic turbidimetric limulus test

    Institute of Scientific and Technical Information of China (English)

    盛凤仙; 熊可强; 王惠之; 张幼捷; 高莉萍

    2012-01-01

    目的 应用动态浊度法定量测定人凝血因子Ⅷ制品中细菌内毒素含量.方法 通过对样品中定量添加标准内毒素的干扰试验,检测其回收率应在50%~200%范围.结果 将样品进行1→100稀释可以有效地消除其对鲎试验的干扰.结论 应用动态浊度法定量测定人凝血因子Ⅷ制品中细菌内毒素含量是可行的,与热原检查法结果一致.%Objective To establish a method for determination of bacterial endotoxin in human coagulation factor Ⅷ by kinetic turbidimetric limulus test. Methods Based on interference experiment, the endotoxin was added in the samples, whose recovery should be within the range 50% - 200% . Results The interference from limulus test was eliminated by diluting the Human coagulation factor Ⅷ (1—100 solution). Conclusions Turbidimetric-kinetic method was feasible, and had the same result with the rabbit-pyretogen test.

  20. The Antagonistic Action of Heat-Clearing and Detoxifying Chinese Drugs on Endotoxins

    Institute of Scientific and Technical Information of China (English)

    张霞; 崔乃杰; 王家泰

    2001-01-01

    @@In the recent decade, interest in treatment and prevention of many critical, severe and acute diseases caused by bacterial endotoxins has been aroused along with the advance of the knowledge on the nature of the endotoxin and the conditions involved. In abroad, attention has been mainly payed to raising antisera and monocolonal antibodies against the endotoxin and the induced mediators. However, the allergic reactions and the cost are still the problems. Till now, there is no drug that can antagonize endotoxin with high effectiveness and low toxicity. Clinical treatments are still confined in inhibiting or killing the pathogen, and correcting the internal environmental disturbance. Being less toxic and rich in resources with low cost and less side-effects, screening of effective Chinese drugs for antagonizing endotoxin is of important and practical significance. Endotoxin belongs to the category of toxic evils, or more precisely, the heat toxin in TCM. Therefore the application of heat-clearing and detoxifying Chinese drugs to antagonizing endotoxins is consistent with the theory of TCM. Some achievements in this field are reported as follows.

  1. Prolastin, a pharmaceutical preparation of purified human α1-antitrypsin, blocks endotoxin-mediated cytokine release

    Directory of Open Access Journals (Sweden)

    Westin Ulla

    2005-01-01

    Full Text Available Abstract Background α1-antitrypsin (AAT serves primarily as an inhibitor of the elastin degrading proteases, neutrophil elastase and proteinase 3. There is ample clinical evidence that inherited severe AAT deficiency predisposes to chronic obstructive pulmonary disease. Augmentation therapy for AAT deficiency has been available for many years, but to date no sufficient data exist to demonstrate its efficacy. There is increasing evidence that AAT is able to exert effects other than protease inhibition. We investigated whether Prolastin, a preparation of purified pooled human AAT used for augmentation therapy, exhibits anti-bacterial effects. Methods Human monocytes and neutrophils were isolated from buffy coats or whole peripheral blood by the Ficoll-Hypaque procedure. Cells were stimulated with lipopolysaccharide (LPS or zymosan, either alone or in combination with Prolastin, native AAT or polymerised AAT for 18 h, and analysed to determine the release of TNFα, IL-1β and IL-8. At 2-week intervals, seven subjects were submitted to a nasal challenge with sterile saline, LPS (25 μg and LPS-Prolastin combination. The concentration of IL-8 was analysed in nasal lavages performed before, and 2, 6 and 24 h after the challenge. Results In vitro, Prolastin showed a concentration-dependent (0.5 to 16 mg/ml inhibition of endotoxin-stimulated TNFα and IL-1β release from monocytes and IL-8 release from neutrophils. At 8 and 16 mg/ml the inhibitory effects of Prolastin appeared to be maximal for neutrophil IL-8 release (5.3-fold, p Conclusion Our data demonstrate for the first time that Prolastin inhibits bacterial endotoxin-induced pro-inflammatory responses in vitro and in vivo, and provide scientific bases to explore new Prolastin-based therapies for individuals with inherited AAT deficiency, but also for other clinical conditions.

  2. Chasing a ghost?--Issues with the determination of circulating levels of endotoxin in human blood.

    Science.gov (United States)

    Gnauck, Anne; Lentle, Roger Graham; Kruger, Marlena Cathorina

    2016-01-01

    Reliable quantification of bacterial products such as endotoxin is important for the diagnosis of Gram-negative infection and for the monitoring of its treatment. Further, it is important to identify patients with persistent subclinical level of bacterial products in their systemic circulation as data from animal studies also suggest this may be correlated with the onset of metabolic syndrome. In this review, we first aim to describe the principles of the Limulus amoebocyte lysate (LAL) test, an assay that is used to quantify endotoxin, and the various shortcomings that must be addressed before it can become a reliable means of quantifying endotoxin in samples derived from blood. We then review published data regarding endotoxin levels in healthy subjects and those with sepsis, inflammatory bowel disease, liver disorders and metabolic disorders such as obesity and diabetes. We also review the evidence regarding influence of macronutrients in augmenting the levels of systemic endotoxin. The results of this review show that reported mean levels of endotoxin in the systemic circulation of healthy humans and of those with various clinical disorders vary over a wide range. Further, this review shows that a significant proportion of this variation can be related to the method that was used to prepare plasma and serum samples prior to assay and its ability to reduce the effect of various blood borne factors that interfere with the LAL assay. PMID:26732012

  3. 21 CFR 866.3210 - Endotoxin assay.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Endotoxin assay. 866.3210 Section 866.3210 Food... DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Serological Reagents § 866.3210 Endotoxin assay. (a) Identification. An endotoxin assay is a device that uses serological techniques in whole blood. The device...

  4. Predictors of airborne endotoxin in the home.

    OpenAIRE

    Park, J. H.; Spiegelman, D L; Gold, D R; Burge, H A; Milton, D K

    2001-01-01

    We identified home characteristics associated with the level of airborne endotoxin in 111 Boston-area homes enrolled in a cohort study of home exposures and childhood asthma, and we developed a predictive model to estimate airborne endotoxin. We measured endotoxin in family-room air and in dust from the baby's bed, family room, bedroom, and kitchen floor. Level of airborne endotoxin was weakly correlated (r < 0.3) with level of endotoxin in each of the four types of dust samples and was signi...

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

    Directory of Open Access Journals (Sweden)

    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

  6. VERTICAL HEREDITY VS. HORIZONTAL GENE TRANSFER: A CHALLENGE TO BACTERIAL CLASSIFICATION

    Institute of Scientific and Technical Information of China (English)

    HAO Bailin; QI Ji

    2003-01-01

    The diversity and classification of microbes has been a long-standing issue. Molecular phylogeny of the prokaryotes based on comparison of the 16S rRNA sequences of the small ribosomal subunit has led to a reasonable tree of life in the late 1970s. However, the availability of more and more complete bacterial genomes has brought about complications instead of refinement of the tree. In particular, it turns out that different choice of genes may tell different history. This might be caused by possible horizontal gene transfer (HGT) among species. There is an urgent need to develop phylogenetic methods that make use of whole genome data. We describe a new approach in molecular phylogeny, namely, tree construction based on K-tuple frequency analysis of the genomic sequences. Putting aside the technicalities, we emphasize the transition from randomness to determinism when the string length K increases and try to comment on the challenge mentioned in the title.

  7. Architecture and signal transduction mechanism of the bacterial chemosensory array: progress, controversies, and challenges.

    Science.gov (United States)

    Falke, Joseph J; Piasta, Kene N

    2014-12-01

    Recent research has deepened our understanding of the ancient, conserved chemosensory array that detects small molecule attractants and repellents, and directs the chemotaxis of bacterial and archaeal cells towards an optimal chemical environment. Here we review advances towards a molecular description of the ultrastable lattice architecture and ultrasensitive signal transduction mechanism of the chemosensory array, as well as controversies and challenges requiring further research. Ultimately, a full molecular understanding of array structure and on-off switching will foster (i) the design of novel therapies that block pathogenic wound seeking and infection, (ii) the development of highly specific, sensitive, stable biosensors, and (iii) the elucidation of general functional principles shared by receptor patches in all branches of life.

  8. Prenatal immune challenge alters the hypothalamic-pituitary-adrenocortical axis in adult rats.

    OpenAIRE

    Reul, J M; Stec, I; Wiegers, G J; Labeur, M S; Linthorst, A C; Arzt, E; Holsboer, F

    1994-01-01

    We investigated whether non-abortive maternal infections would compromise fetal brain development and alter hypothalamic-pituitary-adrenocortical (HPA) axis functioning when adult. To study putative teratogenic effects of a T cell-mediated immune response versus an endotoxic challenge, 10-d-pregnant rats received a single intraperitoneal injection of 5 x 10(8) human red blood cells (HRBC) or gram-negative bacterial endotoxin (Escherichia coli LPS: 30 micrograms/kg). The adult male progeny (3 ...

  9. Single-dose tumor necrosis factor protection against endotoxin-induced shock and tissue injury in rats.

    OpenAIRE

    Alexander, H R; Doherty, G M; Block, M. I.; Kragel, P J; Jensen, J C; Langstein, H N; Walker, E; Norton, J A

    1991-01-01

    Tumor necrosis factor (TNF), a macrophage product released in response to endotoxin and other stimuli, has been shown to be a central mediator of endotoxin or septic shock. However, its highly conserved and wide-ranging physiological effects suggest that it may also be an essential cytokine in the host defense against acute bacterial infection or sepsis. A single nontoxic dose of human recombinant TNF administered intravenously 24 h prior to a lethal infusion of Escherichia coli lipopolysacch...

  10. Structural and functional features of self-assembling protein nanoparticles produced in endotoxin-free Escherichia coli

    OpenAIRE

    Rueda, Fabián; Céspedes, María Virtudes; Sánchez-Chardi, Alejandro; Seras-Franzoso, Joaquin; Pesarrodona, Mireia; Ferrer-Miralles, Neus; Vázquez, Esther; Rinas, Ursula; Unzueta, Ugutz; Mamat, Uwe; Mangues, Ramón; García-Fruitós, Elena; Villaverde, Antonio

    2016-01-01

    Background Production of recombinant drugs in process-friendly endotoxin-free bacterial factories targets to a lessened complexity of the purification process combined with minimized biological hazards during product application. The development of nanostructured recombinant materials in innovative nanomedical activities expands such a need beyond plain functional polypeptides to complex protein assemblies. While Escherichia coli has been recently modified for the production of endotoxin-free...

  11. 凝胶法和动态浊度法检测注射用重组人睫状神经营养因子中细菌内毒素含量%Development of bacterial endotoxin test for recombinant human ciliary neurotrophic factor for injection

    Institute of Scientific and Technical Information of China (English)

    杨军; 应莲芳; 蒋琳; 高雪峰

    2013-01-01

    Objective To develop a method in detection of bacteria endotoxin for recombinant human ciliary neurotrophic factor for injection with the gel-clot test and the kinetic turbidimetric test for the control of the quality of the drug and pre-vention of pyroten reaction in clinical practice .Methods the experiment was conducted according to BET in China Phar-macopoeia 2010 , third section .Results The protein concentration was 100 μg/mL, the detection limit for acterial endo-toxin as defined 10 EU/mL.With the gel-clot test and the kinetic turbidimetric test , results of the tested samples were vali-dated.Conclusion Both the gel-clot test and the kinetic turbidimetric test are suitable for bacterial endotoxin test of re-combinant human ciliary neurotrophic factor for injection .%目的采用凝胶法和动态浊度法检测注射用重组人睫状神经营养因子( CNTF )中细菌内毒素含量,控制制品质量,消除临床热原反应的发生。方法供试品参照《中华人民共和国药典》(2010年版)三部附录ⅫE细菌内毒素检查法中的凝胶法和动态浊度法要求进行检查。结果注射用重组人睫状神经营养因子溶液在100μg/mL质量浓度下,确定内毒素限值为10 EU/mL。凝胶法和动态浊度法检查后,注射用重组人睫状神经营养因子内毒素含量均符合规定。结论凝胶法和动态浊度法可用于注射用重组人睫状神经营养因子的细菌内毒素检查。

  12. Modeling human health risks of airborne endotoxin in homes during the winter and summer seasons

    International Nuclear Information System (INIS)

    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.

  13. 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.

  14. Chemokine-Releasing Microparticles Improve Bacterial Clearance and Survival of Anthrax Spore-Challenged Mice

    Science.gov (United States)

    Espina, Virginia; Liotta, Lance A.; Popov, Serguei G.

    2016-01-01

    In this study the hydrogel microparticles (MPs) were used to enhance migration of neutrophils in order to improve outcome of anthrax infection in a mouse model. Two MP formulations were tested. In the first one the polyacrylamide gel MPs were chemically coupled with Cibacron Blue (CB) affinity bait. In the second one the bait molecules within the MPs were additionally loaded with neutrophil-attracting chemokines (CKs), human CXCL8 and mouse CCL3. A non-covalent interaction of the bait with the CKs provided their gradual release after administration of the MPs to the host. Mice were challenged into footpads with Bacillus anthracis Sterne spores and given a dose of MPs a few hours before and/or after the spores. Pre-treatment with a single dose of CK-releasing MPs without any additional intervention was able to induce influx of neutrophils to the site of spore inoculation and regional lymph nodes correlating with reduced bacterial burden and decreased inflammatory response in footpads. On average, in two independent experiments, up to 53% of mice survived over 13 days. All control spore-challenged but MP-untreated mice died. The CB-coupled particles were also found to improve survival likely due to the capacity to stimulate release of endogenous CKs, but were less potent at decreasing the inflammatory host response than the CK-releasing MPs. The CK post-treatment did not improve survival compared to the untreated mice which died within 4 to 6 days with a strong inflammation of footpads, indicating quick dissemination of spores though the lymphatics after challenge. This is the first report on the enhanced innate host resistance to anthrax in response to CKs delivered and/or endogenously induced by the MPs. PMID:27632537

  15. Gender Difference in Bacteria Endotoxin-Induced Inflammatory and Anorexic Responses.

    Science.gov (United States)

    Kuo, Shiu-Ming

    2016-01-01

    Inflammation-related anorexic response has been observed in systemic diseases as well as in localized infection and is an important issue in patient care. We tested the hypothesis that upon the same endotoxin exposure, males have more severe inflammatory responses and thus suffer from more negative effect on appetite. Ten-week old male and female mice were compared in their plasma levels of pro-inflammatory cytokines after a body weight-based i.p. injection of bacterial endotoxin lipopolysaccharide. Male mice consistently showed significantly higher levels of IL6 and TNFα than female mice. The difference was observed starting at 3 hours after the systemic endotoxin exposure. It was independent of the level of endotoxin dosage and of the genotype of the anti-inflammatory cytokine, IL10. Interestingly, endotoxin-injected male mice also had significantly higher plasma IL10 levels compared to the female mice. Pre-puberty young mice showed no gender differences in the plasma levels of IL6, TNFα and IL10. Their cytokine levels were mostly between that of the adult males and females. Consistent with the higher inflammatory response in male mice, the endotoxin exposure also led to significantly more appetite loss in male mice at a range of doses in two strains of mice. Saline injection in the absence of endotoxin affected neither the cytokine levels nor the appetite. Although a direct mechanistic link between inflammation parameters and appetite was not addressed here, the results support that male gender could be a risk factor for higher pro-inflammatory cytokines and anorexic response after the endotoxin exposure. PMID:27631979

  16. The effect of refrigeration and mixing on detection of endotoxin in parenteral drugs using the Limulus Amebocyte Lysate (LAL) test.

    Science.gov (United States)

    Guilfoyle, D E; Yager, J F; Carito, S L

    1989-01-01

    Prior to testing for the presence of bacterial endotoxin, parenteral products are handled and stored in a variety of ways. Two incidents, detected by the U.S. Food and Drug Administration, revealed that differences in product handling and storage may have played a role in causing analytical discrepancies in the testing of identical samples. The testing procedure was the USP Bacterial Endotoxin test using Limulus Amebocyte Lysate (LAL) reagent. Consequently, an evaluation was made at the two principal factors that contributed to the suspected analytical anomaly. The factors were sample storage and the degree of agitation prior to sample analysis. Additional variables such as bacterial growth medium and adsorption potential of endotoxin by rubber stoppers were also evaluated. It was found that neither the medium employed to grow the E. coli endotoxin nor the storage temperature of the spiked solutions were problematic. However, it was shown that 20-40% of the spiked endotoxin was lost due to non-agitation of solution in vials in which the solution was in contact with the rubber stoppers. A suggested remedy for this problem is to store intact product containers in an upright position and to establish a uniform mixing procedure prior to endotoxin assay.

  17. 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.

  18. Phosphatidylcholine Reverses Ethanol-Induced Increase in Transepithelial Endotoxin Permeability and Abolishes Transepithelial Leukocyte Activation

    DEFF Research Database (Denmark)

    Mitzscherling, Katja; Volynets, Valentina; Parlesak, Alexandr

    2009-01-01

    Chronic alcohol abuse increases both intestinal bacterial overgrowth and intestinal permeability to macromolecules. Intestinal permeability of endotoxin, a component of the outer cell membrane of Gram-negative bacteria, plays a crucial role in the development of alcohol-induced liver disease (ALD...

  19. ADSORBENTS USED IN THE CLEARANCE OF ENDOTOXIN

    Institute of Scientific and Technical Information of China (English)

    YU Mei; LIU Tao; Hou Guanghui; YUAN Zhi

    2003-01-01

    A series of modified poly (methyl methacrylate, PMMA) resins were prepared and compared their adsorption abilities to endotoxin. The results showed that adsorbents, which were grafted with tertiary amine and long spacing arms, had the best adsorption capacities and good blood compatibility, It is hopeful to be used as adsorbent in hemoperfusion for clinical clearance of endotoxin. The influence of original concentration of endotoxin on adsorption and the adsorption mechanism were also investigated.

  20. Endotoxin removal by charge-modified filters.

    OpenAIRE

    Gerba, C P; Hou, K

    1985-01-01

    The effects of positively charged nylon and depth (cellulose-diatomaceous earth) filters on endotoxin removal from various solutions were evaluated. The charged filter media removed significant amounts of Escherichia coli and natural endotoxin from tap water, distilled water, sugars, and NaCl solutions; no significant removal of endotoxin was observed with negatively charged filter media. The extent of removal was influenced by pH, the presence of salts, and organic matter. Such media may be ...

  1. Low dose endotoxin priming is accountable for coagulation abnormalities and organ damage observed in the Shwartzman reaction. A comparison between a single-dose endotoxemia model and a double-hit endotoxin-induced Shwartzman reaction

    Directory of Open Access Journals (Sweden)

    Cate Hugo

    2006-08-01

    Full Text Available Abstract The clinical response of sepsis to a systemic inflammatory infection may be complicated by disseminated intravascular coagulation or DIC. In order to experimentally study the syndrome of DIC, we aimed for a severe sepsis model complicated by disseminated coagulation. Most -simplified- experimental models describing coagulation abnormalities as a consequence of sepsis are based on single dose endotoxemia. The so called-Shwartzman reaction contrarily, is elicited by a low dose endotoxin priming followed by an LPS challenge and is characterized by pathological manifestations that represent the syndrome of DIC. In order to investigate whether the Shwartzman reaction is superior to a single endotoxin challenge as a model for sepsis-induced DIC and to determine what the pathological effect is of an encounter of low endotoxin prior to an LPS challenge, we undertook the present study. In this study we demonstrate that low-dose endotoxin priming prior to an LPS challenge in the Shwartzman reaction is accountable for micro-vascular thrombosis in lung and liver and subsequent (multi- organ failure, not observed after a single-dose endotoxin challenge, which indicates that the Shwartzman reaction is well suited-model to study sepsis-induced DIC adversities. Remarkably, only minor differences in the innate immune response were established between the single-dose endotoxin challenge and the Shwartzman reaction.

  2. Nitric oxide production in celomocytes of the earthworm Eisenia hortensis following bacterial challenge

    Directory of Open Access Journals (Sweden)

    SR Cook

    2015-02-01

    Full Text Available In this in vitro investigation, nitric oxide (NO production was induced within celomocytes of the earthworm Eisenia hortensis following microbial challenge. Celomocytes were pre-loaded with the fluorescent indicator 4-amino-5-methylamino-2’, 7’-difluorofluorescein diacetate (DAF-FM DA in order to detect the presence of intracellular nitric oxide subsequent to a 16 h incubation with chemically-fixed soil bacteria including Bacillus megaterium, Arthrobacter globiformis, Pseudomonas stutzeri, and Azotobacter chroococcum at a range of multiplicities of infection (MOIs. Flow cytometric analysis measuring increases in relative fluorescence intensity (RFI, which is directly proportional to the amount of intracellular NO produced, permitted determination of statistical significance (p < 0.05 of exposed celomocytes compared to baseline controls. Significant increases in NO were detected reproducibly in celomocytes treated with all bacterial species used. The most prominent results were observed after exposure to Gram positive B. megaterium and A. globiformis where 100 % of earthworms tested exhibited statistically significant increases of RFI at MOIs of 100:1 and 500:1, respectively. Furthermore, significant decreases in NO production in bacteria-stimulated earthworm celomocytes incubated with the NOS inhibitor aminoguanidine hydrochloride were observed. These results demonstrate microbial induction of NO synthesis in earthworms and provide evidence of an antimicrobial role of NO in the innate immune system.

  3. 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 sterile water extracts of each device. The main analytical technique for identification of chemical compositions, amounts, and changes during sequential reference Endotoxin additions and subsequent RFGD-treatment removals from infrared (IR)-transparent germanium (Ge) prisms was Multiple Attenuated Internal Reflection (MAIR) infrared spectroscopy sensitive to even monolayer amounts of retained bio-contaminant. KimaxRTM 60 mm x 15 mm and 50mm x 15mm laboratory glass dishes and germanium internal reflection prisms were inoculated with E. coli bacterial endotoxin water suspensions at increments of 0.005, 0.05, 0.5, and 5 EU, and characterized by MAIR-IR spectroscopy of the dried residues on the Ge prisms and LAL Assay of sterile water extracts from both glass and Ge specimens. The Ge prism MAIR-IR measurements

  4. Pubertal immune challenge blocks the ability of estradiol to enhance performance on cognitive tasks in adult female mice

    OpenAIRE

    Ismail, Nafissa; Blaustein, Jeffrey D.

    2012-01-01

    Puberty is a period characterized by brain reorganization that contributes to the development of neural and behavioral responses to gonadal steroids. Previously, we have shown that a single injection of the bacterial endotoxin, lipopolysaccharide (LPS; 1.5mg/kg IP), during the pubertal period (around 6 weeks old) in mice decreases sexual receptivity in response to estradiol and progesterone in adulthood. These findings suggest that pubertal immune challenge has an enduring effect of decreasin...

  5. Evaluation of Endotoxin in Culture Medium for Human in vitro Fertilization

    Institute of Scientific and Technical Information of China (English)

    Jing LI; Wei-jie ZHU; Wen-hong ZHANG

    2005-01-01

    Objective To determine whether the presence of bacterial endotoxin in the commercial culture media utilized for human in vitro fertilization (IVF), and evaluate the difference in detecting endotoxin in culture medium between the human sperm motility assay and the 2-cell mouse embryo assay.Methods Thirty-six batches of culture media commonly used in IVF laboratories from 3 manufacturers were determined for the presence ofendotoxin before using the medium for the assisted reproductive programs (group A). After being used, 25 specimens among above media were also tested (group B). The chromogenic limulus amoebocyte lysate (LAL) test was used for quantification the content of endotoxin. In addition, the human sperm motility assay was compared with the 2-cell mouse embryo assay to evaluate the difference in detecting endotoxin in culture medium.Results Endotoxin was not detected in group A. However, 2 samples were positive in group B. Sperm did not show significant change in motility in group A during 24 h of incubation when compared with the control (P>0.05). However, in group A the 2-cell embryo development to blastocyst was suppressed in 3 batches of media.Conclusions Regular screening of each batch of culture medium should be performed if possible although there was no evidence of endotoxin contamination in commercially prepared pre-tested media. Culture environment should be stringently controlled in case the medium is polluted. The sensitivity of the sperm motility assay was lower than that of the mouse embryo assay for detecting low levels of endotoxin or toxic compounds in the medium.

  6. Comparative Study of Plasma Endotoxin with Procalcitonin Levels in Diagnosis of Bacteremia in Intensive Care Unit Patients

    Science.gov (United States)

    Wang, Tao; Cui, Yun-Liang; Lin, Zhao-Fen; Chen, De-Chang

    2016-01-01

    Background: Both procalcitonin (PCT) and plasma endotoxin levels cannot be solely used for a definite diagnosis of bacteremia or sepsis, and there has been few study comparing the values of the two biomarkers for the diagnosis of bacteremia. The aim of this study was to identify bacteria causing bacteremia and evaluate the role of the two biomarkers in the diagnosis of bacteremia in Intensive Care Unit (ICU). Methods: The medical records of 420 patients in ICU were retrospectively reviewed. Patients (n = 241) who met the inclusion criteria were subjected to blood culture (BC) for the analysis of the endotoxin or PCT levels. The exclusion criteria included the presence of infection with human immunodeficiency virus and/or AIDS, neutropenia without sepsis, pregnancy, treatment with immunosuppressive therapies, or blood diseases such as hematological tumors. Patients’ BC episodes were divided into BC negative, Gram-negative (GN) bacteria, Gram-positive bacteria, and fungi groups. The PCT and plasma endotoxin levels were compared in the different groups. Results: A total of 241 patients with 505 episodes of BC were analyzed. The GN bacteria group showed higher levels of PCT and endotoxin than the BC negative, Gram-positive bacteria, and fungi groups. GN bacteremia was more prevalent than Gram-positive bacteremia. The GN bacteremia caused by non-Enterobacteriaceae infection presented higher endotoxin level than that by Enterobacteriaceae, but no significant difference in PCT levels was observed between the two groups. The plasma endotoxin significantly differed among different groups and was bacterial species dependent. 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

  7. Comparative Study of Plasma Endotoxin with Procalcitonin Levels in Diagnosis of Bacteremia in Intensive Care Unit Patients

    Institute of Scientific and Technical Information of China (English)

    Tao Wang; Yun-Liang Cui; Zhao-Fen Lin; De-Chang Chen

    2016-01-01

    Background: Both procalcitonin (PCT) and plasma endotoxin levels cannot be solely used for a definite diagnosis ofbacteremia or sepsis, and there has been few study comparing the values of the two biomarkers for the diagnosis of bacteremia.The aim of this study was to identify bacteria causing bacteremia and evaluate the role of the two biomarkers in the diagnosis ofbacteremia in Intensive Care Unit (ICU).Methods: The medical records of 420 patients in ICU were retrospectively reviewed.Patients (n =241) who met the inclusion criteria were subjected to blood culture (BC) for the analysis of the endotoxin or PCT levels.The exclusion criteria included the presence of infection with human immunodeficiency virus and/or AIDS, neutropenia without sepsis, pregnancy, treatment with immunosuppressive therapies, or blood diseases such as hematological tumors.Patients' BC episodes were divided into BC negative, Gram-negative (GN) bacteria, Gram-positive bacteria, and fungi groups.The PCT and plasma endotoxin levels were compared in the different groups.Results: A total of 241 patients with 505 episodes of BC were analyzed.The GN bacteria group showed higher levels of PCT and endotoxin than the BC negative, Gram-positive bacteria, and fungi groups.GN bacteremia was more prevalent than Gram-positive bacteremia.The GN bacteremia caused by non-Enterobacteriaceae infection presented higher endotoxin level than that by Enterobacteriaceae, but no significant difference in PCT levels was observed between the two groups.The plasma endotoxin significantly differed among different groups and was bacterial species dependent.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

  8. A probabilistic approach to quantitatively assess the inhalation risk for airborne endotoxin in cotton textile workers

    International Nuclear Information System (INIS)

    Endotoxin, a component of gram-negative bacterial cell walls, is a proinflammatory 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 were carried out to quantitatively assess the potential inhalation risk of airborne endotoxin for workers in cotton textile plants. Combining empirical data and modeling results, we show that the half-maximum effects of the endotoxin dose (ED50) were estimated to be 3.3 x 105 (95% confidence interval (CI): 1.9-14.7 x 105) endotoxin units (EU) for the blood C-reactive protein (CRP) concentration, 1.1 x 105 (95% CI: 0.6-1.7 x 105) EU for the blood polymorphonuclear neutrophil (PMN) count, and 1.5 x 105 (95% CI: 0.4-2.5 x 105) EU for the sputum PMN count. Our study offers a risk-management framework for discussing future establishment of limits for respiratory exposure to airborne endotoxin for workers in cotton textile plants.

  9. A probabilistic approach to quantitatively assess the inhalation risk for airborne endotoxin in cotton textile workers

    Energy Technology Data Exchange (ETDEWEB)

    Liao, Vivian Hsiu-Chuan, E-mail: vivianliao@ntu.edu.tw [Department of Bioenvironmental Systems Engineering, National Taiwan University, 1 Roosevelt Road, Sec. 4, Taipei 106, Taiwan (China); Chou, Wei-Chun; Chio, Chia-Pin; Ju, Yun-Ru; Liao, Chung-Min [Department of Bioenvironmental Systems Engineering, National Taiwan University, 1 Roosevelt Road, Sec. 4, Taipei 106, Taiwan (China)

    2010-05-15

    Endotoxin, a component of gram-negative bacterial cell walls, is a proinflammatory 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 were carried out to quantitatively assess the potential inhalation risk of airborne endotoxin for workers in cotton textile plants. Combining empirical data and modeling results, we show that the half-maximum effects of the endotoxin dose (ED50) were estimated to be 3.3 x 10{sup 5} (95% confidence interval (CI): 1.9-14.7 x 10{sup 5}) endotoxin units (EU) for the blood C-reactive protein (CRP) concentration, 1.1 x 10{sup 5} (95% CI: 0.6-1.7 x 10{sup 5}) EU for the blood polymorphonuclear neutrophil (PMN) count, and 1.5 x 10{sup 5} (95% CI: 0.4-2.5 x 10{sup 5}) EU for the sputum PMN count. Our study offers a risk-management framework for discussing future establishment of limits for respiratory exposure to airborne endotoxin for workers in cotton textile plants.

  10. 显色基质法检测乳酸环丙沙星氯化钠注射液的细菌内毒素%Bacterial endotoxin test of Ciprofloxacin Lactate and Sodium Chloride Injection by chromogenic substrate limulus amebocytelysate

    Institute of Scientific and Technical Information of China (English)

    曾云胜; 张玲莉; 彭燕

    2015-01-01

    Objective To compare bacterial endotoxin test of Ciprofloxacin Lactate and Sodium Chloride Injection by chromogenic substrate limulus amebocytelysate method and sol-gel method. Methods Sample dilution method was used to study of chromogenic substrate limulus reagent on different batches of Ciprofloxacin Lactate and Sodium Chloride Injection, compared with interfering test of sol-gel limulus reagent sample. Results The standard curve correlation co-efficient of chromogenic substrate limulus amebocytelysate was above 0.98. Ciprofloxacin Lactate and Sodium Chloride Injection was not interfere with the concentration of 0.5 mg/mL, which was obviously higher than that of gel colt method (0.125 mg/mL). Conclusion Compared with sol-gel method, chromogenic substrate method has the characteristic of sen-sitive, rapid, quantitative and good repeatability, can be used for content determination on endotoxin of Ciprofloxacin Lactate and Sodium Chloride Injection.%目的:比较显色基质法与凝胶法检测乳酸环丙沙星氯化钠注射液的细菌内毒素的效果。方法采用样品稀释法研究显色基质鲎试剂对不同批号乳酸环丙沙星氯化钠注射液的干扰,并与凝胶法鲎试剂对样品的干扰试验比较。结果显色基质法标准曲线的线性相关系数≥0.98,乳酸环丙沙星氯化钠注射液在0.5 mg/mL浓度时无干扰作用,明显高于凝胶法无干扰浓度(0.125 mg/mL)。结论与凝胶法比较,显色基质法有灵敏、快速、能定量、重复性好的特点,可用于乳酸环丙沙星氯化钠注射液内毒素含量的测定。

  11. Molecular characterization of two novel molecular chaperones in bacterial-challenged Apostichopus japonicus.

    Science.gov (United States)

    Wang, Haihong; Shao, Yina; Zhang, Weiwei; Li, Chenghua; Lv, Zhimeng; Jin, Chunhua

    2015-10-01

    Molecular chaperones of 78 kDa glucose-regulated protein (GRP78) and protein disulfide isomerase (PDI) are involved in protein folding and assembly in the endoplasmic reticulum (ER). Increasing evidences also suggest that these two molecules play an important role in immune response. In the present study, we cloned and characterized GRP78 and PDI genes from Apostichopus japonicus by RNA-seq and RACE approaches (designated as AjGRP78 and AjPDI, respectively). The AjGRP78 cDNA was of 2355bp including an open reading frame (ORF) of 2013 bp encoding a protein of 670 amino acids with three heat shock protein 70 (HSP70) family signatures. AjGRP78 contained a 23-amino acid signal peptide at the N-terminus and a HDEL motif at the C-terminus, which supported the location of the protein in the ER. The full length cDNA of AjPDI was of 1893 bp with a 5' untranslated region (UTR) of 153 bp, a 3' UTR of 228 bp and an ORF of 1512 bp encoding a protein of 503 amino acids. A 17-amino acid signal peptide, two thioredoxin domains with two active sites of CGHC, and KDEL retention signal were totally conserved in the deduced amino acid of AjPDI. Phylogenic analysis and multiple alignments have shown that both genes shared remarkably higher degree of structural conservation and sequence identities with other counterparts from invertebrates and vertebrates, further supporting that the two proteins were novel members of molecular chaperone family. Spatial expression analysis revealed that AjGRP78 mRNA transcripts were dominantly expressed in the tentacle, while AjPDI mRNA levels were abundant in the muscle, intestine and respiratory trees. For Vibrio splendidus challenged sea cucumber, the peak expression of AjGRP78 and AjPDI mRNAs in coelomocytes were detected at 24h with 1.73-fold increase and at 6h with 1.83-fold increase compared with the control group, respectively. Similarly, a significant increase in the relative mRNA levels of AjGRP78 and AjPDI was also identified in 1 μg mL(-1

  12. 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.)

  13. 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.

  14. Endotoxin Induces Fibrosis in Vascular Endothelial Cells through a Mechanism Dependent on Transient Receptor Protein Melastatin 7 Activity

    Science.gov (United States)

    Echeverría, Cesar; Montorfano, Ignacio; Hermosilla, Tamara; Armisén, Ricardo; Velásquez, Luis A.; Cabello-Verrugio, Claudio; Varela, Diego; Simon, Felipe

    2014-01-01

    The pathogenesis of systemic inflammatory diseases, including endotoxemia-derived sepsis syndrome, is characterized by endothelial dysfunction. It has been demonstrated that the endotoxin lipopolysaccharide (LPS) induces the conversion of endothelial cells (ECs) into activated fibroblasts through endothelial­to­mesenchymal transition mechanism. Fibrogenesis is highly dependent on intracellular Ca2+ concentration increases through the participation of calcium channels. However, the specific molecular identity of the calcium channel that mediates the Ca2+ influx during endotoxin-induced endothelial fibrosis is still unknown. Transient receptor potential melastatin 7 (TRPM7) is a calcium channel that is expressed in many cell types, including ECs. TRPM7 is involved in a number of crucial processes such as the conversion of fibroblasts into activated fibroblasts, or myofibroblasts, being responsible for the development of several characteristics of them. However, the role of the TRPM7 ion channel in endotoxin-induced endothelial fibrosis is unknown. Thus, our aim was to study whether the TRPM7 calcium channel participates in endotoxin-induced endothelial fibrosis. Using primary cultures of ECs, we demonstrated that TRPM7 is a crucial protein involved in endotoxin-induced endothelial fibrosis. Suppression of TRPM7 expression protected ECs from the fibrogenic process stimulated by endotoxin. Downregulation of TRPM7 prevented the endotoxin-induced endothelial markers decrease and fibrotic genes increase in ECs. In addition, TRPM7 downregulation abolished the endotoxin-induced increase in ECM proteins in ECs. Furthermore, we showed that intracellular Ca2+ levels were greatly increased upon LPS challenge in a mechanism dependent on TRPM7 expression. These results demonstrate that TRPM7 is a key protein involved in the mechanism underlying endotoxin-induced endothelial fibrosis. PMID:24710004

  15. PREVALENCE OF BACTERIAL RESISTANCE IN HOSPITALIZED CIRRHOTIC PATIENTS IN SOUTHERN BRAZIL: A NEW CHALLENGE

    Directory of Open Access Journals (Sweden)

    Ane Micheli COSTABEBER

    2016-01-01

    Full Text Available Background & Aims: An increased frequency of infections by multiresistant bacteria has been described in hospitalized patients. The aim of this study was to evaluate the bacterial resistance profile in cirrhotic patients. Methods: This is a retrospective observational study. We assessed the antimicrobial susceptibility of 5,839 bacterial isolates from patients with and without cirrhosis. Regarding the multidrug resistance, we evaluated 4,505 bacterial isolates from 2,180 patients. Results: Two hundred and fifty-one patients had cirrhosis (mean age 57.6 ± 11 years; 61.8% were male, 47.8% of cases associated with hepatitis C virus. Of the isolates of patients with and without cirrhosis, 174/464 (37.5% and 1,783/4,041 (44.1% were multiresistant, respectively (p = 0.007. E. coli was the most common multiresistant bacteria in both groups. Approximately 20% of E. coli and Klebsiella sp. isolates were ESBL-producers and 44% of S. aureus isolates were methicillin-resistant in cirrhotic patients. In cirrhotic patients admitted to the emergency department, hospital ward, and intensive care unit, 28.3%, 50% and 40% had multiresistant isolates, respectively. In patients with and without cirrhosis, 36.2% and 33.5% of isolates were resistant to third-generation cephalosporins, respectively. Conclusions: The empirical treatment of infections in hospitalized patients using broad-spectrum antibiotics should consider the observed pattern of bacterial resistance.

  16. PREVALENCE OF BACTERIAL RESISTANCE IN HOSPITALIZED CIRRHOTIC PATIENTS IN SOUTHERN BRAZIL: A NEW CHALLENGE

    Science.gov (United States)

    COSTABEBER, Ane Micheli; de MATTOS, Angelo Alves; SUKIENNIK, Teresa Cristina Teixeira

    2016-01-01

    Background & Aims: An increased frequency of infections by multiresistant bacteria has been described in hospitalized patients. The aim of this study was to evaluate the bacterial resistance profile in cirrhotic patients. Methods: This is a retrospective observational study. We assessed the antimicrobial susceptibility of 5,839 bacterial isolates from patients with and without cirrhosis. Regarding the multidrug resistance, we evaluated 4,505 bacterial isolates from 2,180 patients. Results: Two hundred and fifty-one patients had cirrhosis (mean age 57.6 ± 11 years; 61.8% were male, 47.8% of cases associated with hepatitis C virus). Of the isolates of patients with and without cirrhosis, 174/464 (37.5%) and 1,783/4,041 (44.1%) were multiresistant, respectively (p = 0.007). E. coli was the most common multiresistant bacteria in both groups. Approximately 20% of E. coli and Klebsiella sp. isolates were ESBL-producers and 44% of S. aureus isolates were methicillin-resistant in cirrhotic patients. In cirrhotic patients admitted to the emergency department, hospital ward, and intensive care unit, 28.3%, 50% and 40% had multiresistant isolates, respectively. In patients with and without cirrhosis, 36.2% and 33.5% of isolates were resistant to third-generation cephalosporins, respectively. Conclusions: The empirical treatment of infections in hospitalized patients using broad-spectrum antibiotics should consider the observed pattern of bacterial resistance. PMID:27253738

  17. Biophysical characterization of the interaction of high-density lipoprotein (HDL) with endotoxins.

    Science.gov (United States)

    Brandenburg, Klaus; Jürgens, Gudrun; Andrä, Jörg; Lindner, Buko; Koch, Michel H J; Blume, Alfred; Garidel, Patrick

    2002-12-01

    The interaction of bacterial endotoxins [lipopolysaccharide (LPS) and the 'endotoxic principle' lipid A], with high-density lipoprotein (HDL) from serum was investigated with a variety of physical techniques and biological assays. HDL exhibited an increase in the gel to liquid crystalline phase transition temperature Tc and a rigidification of the acyl chains of the endotoxins as measured by Fourier-transform infrared spectroscopy and differential scanning calorimetry. The functional groups of the endotoxins interacting with HDL are the phosphates and the diglucosamine backbone. The finding of phosphates as target groups is in accordance to measurements of the electrophoretic mobility showing that the zeta potential decreases from -50 to -60 mV to -20 mV at binding saturation. The importance of the sugar backbone as further target structure is in accordance with the remaining negative potential and competition experiments with polymyxin B (PMB) and phase transition data of the system PMB/dephosphorylated LPS. Furthermore, endotoxin binding to HDL influences the secondary structure of the latter manifesting in a change from a mixed alpha-helical/beta-sheet structure to a predominantly alpha-helical structure. The aggregate structure of the lipid A moiety of the endotoxins as determined by small-angle X-ray scattering shows a change of a unilamellar/inverted cubic into a multilamellar structure in the presence of HDL. Fluorescence resonance energy transfer data indicate an intercalation of pure HDL, and of [LPS]-[HDL] complexes into phospholipid liposomes. Furthermore, HDL may enhance the lipopolysaccharide-binding protein-induced intercalation of LPS into phospholipid liposomes. Parallel to these observations, the LPS-induced cytokine production of human mononuclear cells and the reactivity in the Limulus test are strongly reduced by the addition of HDL. These data allow to develop a model of the [endotoxin]/[HDL] interaction. PMID:12444987

  18. Dynamics of Antagonistic Potency of Rhodobacter capsulatus PG Lipopolysaccharide against Endotoxin-Induced Effects.

    Science.gov (United States)

    Kabanov, D S; Serov, D A; Zubova, S V; Grachev, S V; Prokhorenko, I R

    2016-03-01

    The dynamics of antagonistic potency of lipopolysaccharide (LPS) isolated from Rhodobacter capsulatus PG on the synthesis of proinflammatory (TNF-α, IL-1β, IL-8, IL-6, IFN-γ) and antiinflammatory (IL-10, IL-1Ra) cytokines induced by highly stimulatory endotoxins from Escherichia coli or Salmonella enterica have been studied. Using human whole blood, we have shown that R. capsulatus PG LPS inhibited most pronouncedly the endotoxin-induced synthesis of TNF-α, IL-1β, IL-8, and IL-6 during the first 6 h after endotoxin challenge. Similarly, the endotoxin-induced release of IFN-γ was abolished by R. capsulatus PG LPS as well (24 h). In contrast to the above-mentioned cytokines, the relatively weak antagonistic activity of R. capsulatus PG LPS against endotoxin-triggered production of IL-6 and IL-8 was revealed. Since R. capsulatus PG LPS displays more potent antagonistic activity against deleterious effects of S. enterica LPS than those of E. coli LPS in the cases of such cytokines as IL-1β (6 and 24 h), IL-6 and IL-8 (4 h), we conclude that the effectiveness of protective action of antagonist is mostly determined by the primary lipid A structure of the employed agonist.

  19. Polaprezinc Protects Mice against Endotoxin Shock

    OpenAIRE

    Ohata, Shuzo; Moriyama, Chihiro; Yamashita, Atsushi; Nishida, Tadashi; Kusumoto, Chiaki; Mochida, Shinsuke; Minami, Yukari; Nakada, Junya; Shomori, Kohei; Inagaki, Yoshimi; Ohta, Yoshiji; Matsura, Tatsuya

    2010-01-01

    Polaprezinc (PZ), a chelate compound consisting of zinc and l-carnosine (Car), is an anti-ulcer drug developed in Japan. In the present study, we investigated whether PZ suppresses mortality, pulmonary inflammation, and plasma nitric oxide (NO) and tumor necrosis factor (TNF)-α levels in endotoxin shock mice after peritoneal injection of lipopolysaccharide (LPS), and how PZ protects against LPS-induced endotoxin shock. PZ pretreatment inhibited the decrease in the survival rate of mice after ...

  20. 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

  1. Determination of bacterial endotoxin content of diphtheria toxin mutant CRM 197 by kinetic chromogenic assay%动态显色法检测白喉毒素突变体CRM197的细菌内毒素含量

    Institute of Scientific and Technical Information of China (English)

    邓杰; 张珂; 袁涛; 李春阳

    2013-01-01

    目的 建立检测白喉毒素突变体CRM197细菌内毒素(bacterial endotoxin)含量的动态显色法(kinetic chromogenic assay,KCA).方法 按照《中国药典》三部(2010版)要求,用细菌内毒素检查用水(bacterial endotoxin,BET)稀释细菌内毒素工作标准品制备细菌内毒素标准曲线系列溶液,各浓度均设3个平行孔,分别与动态显色鲎试剂(kinetic chromogenic analysis tachypleus amebocyte lysate,KCA TAL)反应,绘制标准曲线,验证标准曲线的可靠性,确定最佳线性范围、测定范围及最低检测限(limit of quantitation,LOQ),验证方法的精密性和准确性,并进行初步应用.结果 标准曲线的回归方程为:y=-0.263x+2.741,R2=0.997,相关系数的绝对值|r|≥0.980,阴性对照的反应时间大于标准曲线最低点的反应时间,各复孔的变异系数(CV)<10%;内毒素浓度在0.02~2.50EU/ml时,线性关系良好,LOQ为0.02 EU/ml;3个浓度(2.50、0.50、0.02 EU/ml)标准内毒素检测结果的CV均<5%,加入高、中、低3个浓度(2.50、0.50、0.02 EU/ml)标准内毒素的3批供试品检测结果的CV均<10%,回收率在95% ~ 143%之间;采用建立的方法检测10批次CRM197样品的细菌内毒素含量,回收率在77%~118%之间,其中8批样品的内毒素含量合格.结论 动态显色法检测CRM197中内毒素的含量精密性和准确性良好,能快速、定量检测样品中的内毒素含量,抗干扰能力强,可用于CRM197研制过程中的质量控制.

  2. Spontaneous bacterial peritonitis: The clinical challenge ofa leaky gut and a cirrhotic liver

    Institute of Scientific and Technical Information of China (English)

    Philipp Lutz; Hans Dieter Nischalke; Christian P Strassburg; Ulrich Spengler

    2015-01-01

    Spontaneous bacterial peritonitis (SBP) is a frequent,life-threatening bacterial infection in patients withliver cirrhosis and ascites. Portal hypertension leadsto increased bacterial translocation from the intestine.Failure to eliminate invading pathogens due to immunedefects associated with advanced liver disease on thebackground of genetic predisposition may result in SBP.The efficacy of antibiotic treatment and prophylaxis hasdeclined due to the spread of multi-resistant bacteria.Patients with nosocomial SBP and with prior antibiotictreatment are at a particularly high risk for infectionwith resistant bacteria. Therefore, it is important toadapt empirical treatment to these risk factors and tothe local resistance profile. Rifaximin, an oral, nonabsorbableantibiotic, has been proposed to preventSBP, but may be useful only in a subset of patients.Since novel antibiotic classes are lacking, we have todevelop prophylactic strategies which do not inducebacterial resistance. Farnesoid X receptor agonistsmay be a candidate, but so far, clinical studies are notavailable. New diagnostic tests which can be carriedout quickly at the patient's site and provide additionalprognostic information would be helpful. Furthermore,we need tools to predict antibiotic resistance in orderto tailor first-line antibiotic treatment of spontaneousbacterial peritonitis to the individual patient and toreduce mortality.

  3. A cohabitation challenge to compare the efficacies of vaccines for bacterial kidney disease (BKD) in chinook salmon Oncorhynchus tshawytscha

    Science.gov (United States)

    Alcorn, S.; Murray, A.L.; Pascho, R.J.; Varney, J.

    2005-01-01

    The relative efficacies of 1 commercial and 5 experimental vaccines for bacterial kidney disease (BKD) were compared through a cohabitation waterborne challenge. Groups of juvenile chinook salmon Oncorhynchus tshawytscha were vaccinated with one of the following: (1) killed Renibacterium salmoninarum ATCC 33209 (Rs 33209) cells; (2) killed Rs 33209 cells which had been heated to 37??C for 48 h, a process that destroys the p57 protein; (3) killed R. salmoninarum MT239 (Rs MT239) cells; (4) heated Rs MT239 cells; (5) a recombinant version of the p57 protein (r-p57) emulsified in Freund's incomplete adjuvant (FIA); (6) the commercial BKD vaccine Renogen; (7) phosphate-buffered saline (PBS) emulsified with an equal volume of FIA; or (8) PBS alone. Following injection, each fish was marked with a subcutaneous fluorescent latex tag denoting its treatment group and the vaccinated fish were combined into sham and disease challenge tanks. Two weeks after these fish were vaccinated, separate groups of fish were injected with either PBS or live R. salmoninarum GL64 and were placed inside coated-wire mesh cylinders (liveboxes) in the sham and disease challenge tanks, respectively. Mortalities in both tanks were recorded for 285 d. Any mortalities among the livebox fish were replaced with an appropriate cohort (infected with R. salmoninarum or healthy) fish. None of the bacterins evaluated in this study induced protective immunity against the R. salmoninarum shed from the infected livebox fish. The percentage survival within the test groups in the R. salmoninarum challenge tank ranged from 59% (heated Rs MT239 bacterin) to 81 % (PBS emulsified with FIA). There were no differences in the percentage survival among the PBS-, PBS/FIA-, r-p57-and Renogen-injected groups. There also were no differences in survival among the bacterin groups, regardless of whether the bacterial cells had been heated or left untreated prior to injection. ?? Inter-Research 2005.

  4. Isolation of Endotoxin Eliminating Lactic Acid Bacteria and a Property of Endotoxin Eliminating Protein.

    Science.gov (United States)

    Kondo, Ayaka; Asami, Kyoko; Suda, Yoshihito; Shimoyamada, Makoto; Kanauchi, Makoto

    2016-06-01

    Recently, many scholars have reported lactic acid bacteria (LAB) functions, such as anticancer activity and anti-inflammatory activity for intestines. To decrease inflammatory substances such as endotoxins, LAB consumed safely with meals were isolated from food and food ingredients. First, LAB were isolated as 168 strains of bacillus LAB (49 strain) and coccus LAB (119 strains) from food ingredients and fermented foods such as rice, rice bran, malt, grains, miso soy paste, and some pickles. Their LAB (168 strains) were cultivated in medium containing endotoxin from Escherichia coli O18 LPS at 15 and 30 °C for 64 h to identify endotoxin-eliminating LAB. Consequently, the AK-23 strain was screened as an endotoxin-eliminating LAB strain. The strain decreased endotoxin in YP medium without sugar at 30 °C for 64 h until 9% of endotoxin. The strain was identified as Pediococcus pentosaceus according to morphological characteristics such as its cell shape, physiological characteristics related to its fermentation type, assimilation of sugars, pH tolerance, optimum growth temperature, and molecular biological characteristics as its homology to 16S rRNA. To investigate the location of the endotoxin-eliminating substance, 4 fractions were separated from AK-23 cells as extracellular, cell wall digestion, cytoplasm, and cell membrane fractions. The endotoxin-decreasing substance, located on a cell wall, was identified as a 217 kDa protein. PMID:27096744

  5. Challenges of studying viral aerosol metagenomics and communities in comparison with bacterial and fungal aerosols.

    Science.gov (United States)

    Prussin, Aaron J; Marr, Linsey C; Bibby, Kyle J

    2014-08-01

    Despite the obvious importance of viral transmission and ecology to medicine, epidemiology, ecology, agriculture, and microbiology, the study of viral bioaerosols and community structure has remained a vastly underexplored area, due to both unresolved technical challenges and unrecognized importance. High-throughput, culture-independent techniques such as viral metagenomics are beginning to revolutionize the study of viral ecology. With recent developments in viral metagenomics, characterization of viral bioaerosol communities provides an opportunity for high-impact future research. However, there remain significant challenges for the study of viral bioaerosols compared with viruses in other matrices, such as water, the human gut, and soil. Collecting enough biomass is essential for successful metagenomic analysis, but this is a challenge with viral bioaerosols. Herein, we provide a perspective on the importance of studying viral bioaerosols, the challenges of studying viral community structure, and the potential opportunities for improvements in methods to study viruses in indoor and outdoor air.

  6. 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.

  7. Evolution of bacterial meningitis diagnosis in Sao Paulo State-Brazil and future challenges

    Directory of Open Access Journals (Sweden)

    Maristela Marques Salgado

    2013-09-01

    Full Text Available Bacterial meningitis (BM is a severe disease and still represents a serious public health problem with high rates of morbidity and mortality. The most common cases of BM around the world, mainly in Brazil, have been caused by Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae type b. Bacterial culture is the gold-standard technique for BM confirmation, but approximately 50% of suspected cases are not culture-confirmed, due to problems related to improper transportation and seeding or previous antibiotic treatment. Immunological methods present low sensitivity and have possibility of cross-reactions. Real time PCR (qPCR is a molecular technique and has been successful used for BM diagnosis at Instituto Adolfo Lutz in São Paulo State, Brazil, since 2007. The incorporation of qPCR in the Public Health surveillance routine in our state resulted in diminishing 50% of undetermined BM cases. Our efforts are focused on qPCR implementation in the BM diagnostic routine throughout Brazil.

  8. Cytokine release from human peripheral blood leucocytes incubated with endotoxin with and without prior infection with influenza virus

    DEFF Research Database (Denmark)

    Banner, Jytte; Smith, H; Sweet, C

    1993-01-01

    Previous work with a neonatal ferret model for human SIDS had indicated that inflammation caused by a combination of influenza virus and bacterial endotoxin may be a cause of human SIDS. To determine whether cytokines may be involved in this inflammatory response, levels of interleukin (IL)-1 bet...... determined using bioassays rather than ELISA assays, the pattern of increased yields from cells incubated with virus and endotoxin compared with either alone was still evident. The possible importance of these observations for SIDS victims is discussed....

  9. 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. PMID:20434900

  10. Comparison of several control standard endotoxins to the National Reference Standard Endotoxin--an HIMA collaborative study.

    OpenAIRE

    Pearson, F C; Weary, M E; Sargent, H E; Novitsky, T J; Lin, H.; Lindsay, G.; Berzofsky, R N; Lane, A L; Wilson, J. D.; Cooper, J. F.

    1985-01-01

    A collaborative study, initiated under the auspices of the Health Industry Manufacturers Association (HIMA), was designed to establish the relationship of Escherichia coli O55:B5 endotoxin (the control standard endotoxin of HIMA and the Food and Drug Administration's Office of Medical Devices) to the U.S. National Reference Standard Endotoxin and to two internationally used control standard endotoxins. By using two Limulus amoebocyte lysate test systems, it was established that the E. coli O5...

  11. Genome expression analysis of Anopheles gambiae: Responses to injury, bacterial challenge, and malaria infection

    OpenAIRE

    Dimopoulos, George; Christophides, George K.; Meister, Stephan; SCHULTZ, JÖRG; White, Kevin P.; Barillas-Mury, Carolina; Kafatos, Fotis C.

    2002-01-01

    The complex gene expression responses of Anopheles gambiae to microbial and malaria challenges, injury, and oxidative stress (in the mosquito and/or a cultured cell line) were surveyed by using cDNA microarrays constructed from an EST-clone collection. The expression profiles were broadly subdivided into induced and down-regulated gene clusters. Gram+ and Gram− bacteria and microbial elicitors up-regulated a diverse set of genes, many belonging to the immunity class, and the response to malar...

  12. Effects of severe acute pancreatitis on gut bacteria/endotoxin translocation in pigs

    Institute of Scientific and Technical Information of China (English)

    TU Wei-feng; XI Wen-bin; DAI Jian-qiang; HE Jing; LIU Zhong-hua

    2005-01-01

    Objective: To set up a swine model of severe acute pancreatitis(SAP) and to observe its relationship with the gut-originated bacteria/endotoxin translocation. Methods: Forty pigs weighing 17-22 kg were randomly divided into SAP group (n= 34) and sham-SAP group(n= 6). By injecting 1 ml/kg of com bined solution of 5% sodium taurocholate and 8 000-10 000 benzoyl arginine ethyl ester (BAEE) units trypsin per milliliter into pancreas via pancreatic duct, SAP was induced under anesthesia. Endotoxin samples from vena cava were determined by chromogenic limulus amebocyte lysate (LAL) technique. Both portal and central vena blood samples were collected before and 72 h after the induction of SAP and cultured for both aerobic and anaerobic bacterial growth. Animals were sacrificed at the end of experiments by injecting 20 ml of 10% KC1 intravenously and tissue specimens of mesenteriolum and mesocolon lymph nodes, lung, pulmonary portal lymph nods and pancreas were taken immediately after animal death, and homogenized for bacteriological studies. Results: Systemic plasma endotoxin levels increased rapidly 6 h post induction of SAP(PIS) with a peak at 48 h PIS (P<0.01). The magnitude of bacterial translocation in both portal and systemic blood and remote systemic organs as well were recovered PIS. Conclusion:(1)A swine model of SAP was established; (2)The early endotoxemia PIS seamed probable originated from gut endotoxin translocation; (3)The magnitude of bacterial translocation in both portal and systemic blood and the remote systemic organs as well were recovered at 72h PIS.

  13. 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.

  14. Small intestinal bacterial overgrowth in nonalcoholic steatohepatitis: association with toll-like receptor 4 expression and plasma levels of interleukin 8.

    LENUS (Irish Health Repository)

    Shanab, Ahmed Abu

    2011-05-01

    Experimental and clinical studies suggest an association between small intestinal bacterial overgrowth (SIBO) and nonalcoholic steatohepatitis (NASH). Liver injury and fibrosis could be related to exposure to bacterial products of intestinal origin and, most notably, endotoxin, including lipopolysaccharide (LPS).

  15. Exposure to bacterial signals does not alter pea aphids' survival upon a second challenge or investment in production of winged offspring.

    Directory of Open Access Journals (Sweden)

    Bas ter Braak

    Full Text Available Pea aphids have an obligate nutritional symbiosis with the bacteria Buchneraaphidicola and frequently also harbor one or more facultative symbionts. Aphids are also susceptible to bacterial pathogen infections, and it has been suggested that aphids have a limited immune response towards such pathogen infections compared to other, more well-studied insects. However, aphids do possess at least some of the genes known to be involved in bacterial immune responses in other insects, and immune-competent hemocytes. One possibility is that immune priming with microbial elicitors could stimulate immune protection against subsequent bacterial infections, as has been observed in several other insect systems. To address this hypothesis we challenged aphids with bacterial immune elicitors twenty-four hours prior to live bacterial pathogen infections and then compared their survival rates to aphids that were not pre-exposed to bacterial signals. Using two aphid genotypes, we found no evidence for immune protection conferred by immune priming during infections with either Serratia marcescens or with Escherichia coli. Immune priming was not altered by the presence of facultative, beneficial symbionts in the aphids. In the absence of inducible immune protection, aphids may allocate energy towards other defense traits, including production of offspring with wings that could escape deteriorating conditions. To test this, we monitored the ratio of winged to unwinged offspring produced by adult mothers of a single clone that had been exposed to bacterial immune elicitors, to live E. coli infections or to no challenge. We found no correlation between immune challenge and winged offspring production, suggesting that this mechanism of defense, which functions upon exposure to fungal pathogens, is not central to aphid responses to bacterial infections.

  16. 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.

  17. Measurement Uncertainty of Chromogenic LAL Assays: Reaction Time and Proportion of Endotoxin and LAL Reagent Affect Release of p-Nitroaniline.

    Science.gov (United States)

    Ostronoff, Celina Silva; Lourenço, Felipe Rebello

    2015-01-01

    Limulus Amebocyte Lysate (LAL) assays are widely used for detection and quantification of bacterial endotoxins in pharmaceuticals and medical devices. However, there are only a few studies on the measurement uncertainty of LAL assays. The aim of this work was to identify and quantify the main sources of measurement uncertainty for end point and kinetic-chromogenic LAL assays. Response surface methodology was used to study how the release of p-nitroaniline (pNA) is affected by reaction time and proportion of endotoxin and LAL reagent in end point and kinetic-chromogenic LAL assays, respectively. Increased release of pNA was observed when reaction time was increased. In addition, if different volumes of sample (or endotoxin standard) and LAL reagent are used, the pNA release rate will be affected. These results may be due to the increased interaction between the bacterial endotoxin and LAL-activated enzyme. Final measurement uncertainties (95% confidence interval) were 90-120% and 90-127% of bacterial endotoxin content for end point and kinetic-chromogenic assays, respectively. These values are reasonable for the scope of the method and allow the application of these measurement uncertainties in routine analysis of pharmaceuticals and medical devices.

  18. Genome expression analysis of Anopheles gambiae: responses to injury, bacterial challenge, and malaria infection.

    Science.gov (United States)

    Dimopoulos, George; Christophides, George K; Meister, Stephan; Schultz, Jörg; White, Kevin P; Barillas-Mury, Carolina; Kafatos, Fotis C

    2002-06-25

    The complex gene expression responses of Anopheles gambiae to microbial and malaria challenges, injury, and oxidative stress (in the mosquito and/or a cultured cell line) were surveyed by using cDNA microarrays constructed from an EST-clone collection. The expression profiles were broadly subdivided into induced and down-regulated gene clusters. Gram+ and Gram- bacteria and microbial elicitors up-regulated a diverse set of genes, many belonging to the immunity class, and the response to malaria partially overlapped with this response. Oxidative stress activated a distinctive set of genes, mainly implicated in oxidoreductive processes. Injury up- and down-regulated gene clusters also were distinctive, prominently implicating glycolysis-related genes and citric acid cycle/oxidative phosphorylation/redox-mitochondrial functions, respectively. Cross-comparison of in vivo and in vitro responses indicated the existence of tightly coregulated gene groups that may correspond to gene pathways. PMID:12077297

  19. Peptides with dual mode of action: Killing bacteria and preventing endotoxin-induced sepsis.

    Science.gov (United States)

    Brandenburg, Klaus; Heinbockel, Lena; Correa, Wilmar; Lohner, Karl

    2016-05-01

    Bacterial infections, with the most severe form being sepsis, can often not be treated adequately leading to high morbidity and lethality of infected patients in critical care units. In particular, the increase in resistant bacterial strains and the lack of new antibiotics are main reasons for the worsening of the current situation, As a new approach, the use of antimicrobial peptides (AMPs) seems to be promising, combining the ability of broad-spectrum bactericidal activity and low potential of induction of resistance. Peptides based on natural defense proteins or polypeptides such as lactoferrin, Limulus anti-lipopolysaccharide factor (LALF), cathelicidins, and granulysins are candidates due to their high affinity to bacteria and to their pathogenicity factors, in first line lipopolysaccharide (LPS, endotoxin) of Gram-negative origin. In this review, we discuss literature with the focus on the use of AMPs from natural sources and their variants as antibacterial as well as anti-endotoxin (anti-inflammatory) drugs. Considerable progress has been made by the design of new AMPs for acting efficiently against the LPS-induced inflammation reaction in vitro as well as in vivo (mouse) models of sepsis. Furthermore, the data indicate that efficient antibacterial compounds are not necessarily equally efficient as anti-endotoxin drugs and vice versa. The most important reason for this may be the different molecular geometry of LPS in bacteria and in free form. This article is part of a Special Issue entitled: Antimicrobial peptides edited by Karl Lohner and Kai Hilpert. PMID:26801369

  20. Identification and expression analysis of TLR2 in mucosal tissues of turbot (Scophthalmus maximus L.) following bacterial challenge.

    Science.gov (United States)

    Liu, Fengqiao; Su, Baofeng; Gao, Chengbin; Zhou, Shun; Song, Lin; Tan, Fenghua; Dong, Xiaoyu; Ren, Yichao; Li, Chao

    2016-08-01

    The pathogen recognition receptors (PRRs), which can recognize the conserved pathogen-associated molecular patterns (PAMPs) of the bacteria, play key roles in the mucosal surfaces for pathogen recognition and activation of immune signaling pathways. However, our understanding of the PRRs and their activities in mucosal surfaces in the critical early time points during pathogen infection is still limited. Towards to this end, here, we sought to identify the Toll-like receptor 2 (TLR2) in turbot as well as its expression profiles in mucosal barriers following bacterial infection in the early time points. The full-length TLR2 transcript consists of open reading frame (ORF) of 2451 bp encoding the putative peptide of 816 amino acids. The phylogenetic analysis revealed the turbot TLR2 showed the closest relationship to Paralichthys olivaceus. The TLR2 mRNA expression could be detected in all examined tissues, with the most abundant expression level in liver, and the lowest expression level in skin. In addition, TLR2 showed different expression patterns following Vibrio anguillarum and Streptococcus iniae infection, but was up-regulated following both challenge, especially post S. iniae challenge. Characterization of TLR2 will probably contribute to understanding of a number of infectious diseases and broaden the knowledge of interactions between host and pathogen, which will eventually help in the development of novel intervention strategies for farming turbot. PMID:27368539

  1. Decreased mortality of lake michigan chinook salmon after bacterial kidney disease challenge: Evidence for pathogen-driven selection?

    Science.gov (United States)

    Purcell, M.K.; Murray, A.L.; Elz, A.; Park, L.K.; Marcquenski, S.V.; Winton, J.R.; Alcorn, S.W.; Pascho, R.J.; Elliott, D.G.

    2008-01-01

    In the late 1960s, Chinook salmon Oncorhynchus tshawytscha from the Green River, Washington, were successfully introduced into Lake Michigan. During spring from1988 to 1992, large fish die-offs affecting Chinook salmon occurred in the lake. Multiple ecological factors probably contributed to the severity of the fish kills, but the only disease agent found regularly was Renibacterium salmoninarum, the causative agent of bacterial kidney disease. in this study, survival after challenge by R. salmoninarum was compared between two Chinook salmon stocks: a Lake Michigan stock from Wisconsin (WI) and the progenitor stock from the Green River. We found that the WI stock had significantly greater survival than the Green River stock. Next, the WI and Green River stocks were exposed to the marine pathogen Listonella anguillarum (formerly Vibrio anguillarum), one of the causative agents of vibriosis; survival after this challenge was significantly poorer for the WI stock than for the Green River stock. A close genetic relationship between the Green River and WI stocks was confirmed by analyzing 13 microsatellite loci. These results collectively suggest that disease susceptibility of Lake Michigan Chinook salmon has diverged from that of the source population, possibly in response to pathogen-driven selection. ?? Copyright by the American Fisheries Society 2008.

  2. Association of plasma endotoxin, inflammatory cytokines and risk of colorectal adenomas

    International Nuclear Information System (INIS)

    Recent studies suggest that bacterial endotoxins may be associated with various chronic diseases, including colorectal adenomas and cancer. Given the evidence linking inflammation and colorectal cancer, we sought to determine if plasma endotoxin concentrations are associated with indicators of systemic or local inflammation and colorectal adenomas. This cross-sectional study consisted of participants who underwent screening colonoscopies and included adenoma cases (n=138) and non-adenoma controls (n=324). Plasma concentrations of endotoxin were measured with Limulus Amebocyte Lysate (LAL) assay. We quantified concentrations of inflammatory cytokines, interleukin-4 (IL-4), IL-6, IL-8, IL-10, IL-12, tumor necrosis factor-alpha (TNF-α), and interferon-γ (IFN-γ) in plasma by ELISA and mRNA expression levels in rectal mucosal biopsies by quantitative RT-PCR. Interleukin-17 was evaluated only in the rectal mucosa. Compared to subjects with low plasma endotoxin concentrations, those with higher concentrations were more likely to have adenomas (OR 1.4, 95% CI 1.0-2.1). Among subjects with adenomas, those with villous histology were more likely to have higher endotoxin concentrations (5.4 vs. 4.1EU/mL, p=0.05) and lower plasma IFN-γ (0 vs. 1.64 pg/mL, p=0.02) compared to those with only tubular adenomas. Cases showed a trend of having higher plasma TNF-α levels than controls (p=0.06), but none of the other plasma or rectal mucosal cytokine levels differed between cases and controls. Elevated mucosal IL-12 levels were associated with having multiple adenomas (p=0.04). Higher concentrations of plasma endotoxin predicted increased plasma IL-12 levels (OR 1.5, 95% CI 1.0-2.2) and rectal mucosal IL-12 (OR 1.9, 95% CI 1.0-3.7) and IL-17 gene expression (OR 2.2, 95% CI 1.0-4.6). These findings suggest that interactions between elevated plasma endotoxin concentrations and inflammatory cytokines may be relevant to the development of colorectal adenomas

  3. miR-200a-3p regulates TLR1 expression in bacterial challenged miiuy croaker.

    Science.gov (United States)

    Wang, Yanjin; Xu, Guoliang; Han, Jingjing; Xu, Tianjun

    2016-10-01

    MicroRNAs (miRNAs) are highly conserved, small non-coding RNAs which post-transcriptionally regulate various biological processes by repressing mRNA translation or degradating mRNA. It has been demonstrated that miRNAs play crucial roles in regulating the immune system. In this study, we explored the potential roles of miR-200a-3p in regulating TLR signaling pathway in miiuy croaker. Bioinformatics analysis showed that miiuy croaker TLR1 (mmiTLR1) was a putative target of miR-200a-3p. Negative expression profiles in spleen of Vibrio anguillarum challenged miiuy croaker and in lipopolysaccharide (LPS) stimulated miiuy croaker leukocytes further validated the prediction. Luciferase reporter assays showed that the dual-luciferase reporter fused to the 3'UTR of wild type mmiTLR1 cotransfected with miR-200a-3p mimics exhibited a reduction in luciferase activity compared with the controls. All of the present data provide direct evidence that miR-200a-3p is involved in TLR1 expression modulation in miiuy croaker, which will offer a basis for better understanding of miRNA regulation in fish TLR signaling pathways. PMID:27288848

  4. Endotoxin exposure assessment - measurement and characterization

    NARCIS (Netherlands)

    Spaan, S.

    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 fac

  5. Hypertonic saline enhances host response to bacterial challenge by augmenting receptor-independent neutrophil intracellular superoxide formation.

    LENUS (Irish Health Repository)

    Shields, Conor J

    2012-02-03

    OBJECTIVE: This study sought to determine whether hypertonic saline (HTS) infusion modulates the host response to bacterial challenge. METHODS: Sepsis was induced in 30 Balb-C mice by intraperitoneal injection of Escherichia coli (5 x 107 organisms per animal). In 10 mice, resuscitation was performed at 0 and 24 hours with a 4 mL\\/kg bolus of HTS (7.5% NaCl), 10 animals received 4 mL\\/kg of normal saline (0.9% NaCl), and the remaining animals received 30 mL\\/kg of normal saline. Samples of blood, spleen, and lung were cultured at 8 and 36 hours. Polymorphonucleocytes were incubated in isotonic or hypertonic medium before culture with E. coli. Phagocytosis was assessed by flow cytometry, whereas intracellular bacterial killing was measured after inhibition of phagocytosis with cytochalasin B. Intracellular formation of free radicals was assessed by the molecular probe CM-H(2)DCFDA. Mitogen-activated protein (MAP) kinase p38 and ERK-1 phosphorylation, and nuclear factor kappa B (NFkappaB) activation were determined. Data are represented as means (SEM), and an analysis of variance test was performed to gauge statistical significance. RESULTS: Significantly reduced bacterial culture was observed in the animals resuscitated with HTS when compared with their NS counterparts, in blood (51.8 +\\/- 4.3 vs. 82.0 +\\/- 3.3 and 78.4 +\\/- 4.8, P = 0.005), lung (40.0 +\\/- 4.1 vs. 93.2 +\\/- 2.1 and 80.9 +\\/- 4.7, P = 0.002), and spleen (56.4 +\\/- 3.8 vs. 85.4 +\\/- 4.2 and 90.1 +\\/- 5.9, P = 0.05). Intracellular killing of bacteria increased markedly (P = 0.026) and superoxide generation was enhanced upon exposure to HTS (775.78 +\\/- 23.6 vs. 696.57 +\\/- 42.2, P = 0.017) despite inhibition of MAP kinase and NFkappaB activation. CONCLUSIONS: HTS significantly enhances intracellular killing of bacteria while attenuating receptor-mediated activation of proinflammatory cascades.

  6. Role of endotoxin in pathogenesis of hepatocellular carcinoma in mice%内毒素在小鼠肝癌发生过程中的作用

    Institute of Scientific and Technical Information of China (English)

    韩殿冰; 曾本华; 葛成果

    2015-01-01

    Objective To explore the role of endotoxin in HCC induced by the DEN .Methods Four groups of male mice were fed with DEN.Group A is the germ free mice , group B is also the germ free mice injected by endotoxin in abdominal cavity .group C is normal bacte-rial mice, group D is also normal bacterial mice injected by anti -endotoxin IgY in abdominal cavity .Results The highest level of endotoxin in B group and C group had the highest incidence of hepatocellular carcinoma , the lowest level of endotoxin in A group and D group , the incidence of HCC is lowest .Endotoxin levels and the incidence of HCC was positively related .Conclusion Endotoxin is the root cause of liver cancer .%目的 探讨内毒素在二乙基亚硝铵( DEN)诱发小鼠肝癌过程中的作用. 方法 四组雄性小鼠均喂食致癌物DEN液, A组为无菌小鼠,B组无菌小鼠腹腔注射内毒素,C组为普通小鼠,D组普通小鼠腹腔注射抗内毒素IgY. 观察肝癌发生率和血清内毒素水平. 结果 内毒素水平最高的B组和C组肝癌发生率最高,内毒素水平最低的A组和D组肝癌发生率也最低,内毒素水平和肝癌发生率呈显著正相关. 结论 细菌的代谢产物内毒素是引起肝癌的根本原因.

  7. ROLE OF ENDOGENOUS CARBON MONOXIDE IN ENDOTOXIN SHOCK

    Institute of Scientific and Technical Information of China (English)

    史源; 李华强; 潘捷; 覃世文; 潘凤; 蒋东波; 沈际皋

    2000-01-01

    Objective. To study the role of endogenous carbon monoxide (CO) in endotoxin shock. Methods. The changes of CO levels and the effects of zinc protoporphyrin IX (ZnPP), an inhibitor of hemeoxygenase (HO), in endotoxin shock and the efficacy of heroin, an inducer of HO were investigated. Results. The plasma CO levels were found to be significantly increased during the comse of endotoxin shock. Injection of ZnPP was shown to abrogate the endotoxin-induced hypotension and metabolic derangements markedly. Administration of hemin to healthy rabbits revealed the hypotension and metabolic derangements similar to the animsls given endotoxin. Conclusion. CO is a newly found endogenously produced mediator which may play an important role in the pathogenesis of endotoxin shock.

  8. Ultrasensitive detection of endotoxins using computationally designed nanoMIPs

    OpenAIRE

    Altintas, Zeynep; Abdin, Mohammed J.; Tothill, Alexander M.; Karim, Kal; Tothill, Ibtisam E.

    2016-01-01

    Novel molecularly imprinted polymer nanoparticles (nanoMIPs) were designed for endotoxin from Escherichia coli 0111:B4, using computational modeling. The screening process based on binding energy between endotoxin and each monomer was performed with 21 commonly used monomers, resulting in the selection of itaconic acid, methacrylic acid and acrylamide as functional monomers due to their strong binding interaction with the endotoxin template. The nanoMIPs were successfully synthesized with fun...

  9. 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.

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

    International Nuclear Information System (INIS)

    Highlights: ► A HK-2 cells model of inflammation-induced acute kidney injury. ► Two oximetry methods: high resolution respirometry and ESR spectroscopy. ► Oxygen consumption rates of renal cells decrease when treated with LPS. ► Cells do not recover normal respiration when the LPS treatment is removed. ► 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.

  11. 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.

  12. Effect of endotoxin on radiation syndrome - a review

    International Nuclear Information System (INIS)

    A review is given of studies on the effect of endotoxin on postirradiation syndrome. Recent knowledge is summed up on the compositions of endotoxins and on their general biological effects. Endotoxins of different bacteria are discussed such as are utilized for favourably affecting the postirradiation syndrome. For each bacterium the classification is shown in the microbe system. This, however, is not standard in the literature. General assessment is made of studies published so far on radiation protection of organisms with endotoxins. (B.S.)

  13. Bronchial hyperreactivity, increased endotoxin lethality and melanocytic tumorigenesis in transgenic mice overexpressing platelet-activating factor receptor.

    OpenAIRE

    Ishii, S.; Nagase, T; Tashiro, F; Ikuta, K. (Koichi); Sato, S.; Waga, I.; Kume, K.; Miyazaki, J; Shimizu, T

    1997-01-01

    Although platelet-activating factor (PAF) has been shown to exert pleiotropic effects on isolated cells or tissues, controversy still exists as to whether it plays significant pathophysiological roles in vivo. To answer this question, we established transgenic mice over-expressing a guinea-pig PAF receptor (PAFR). The transgenic mice showed a bronchial hyperreactivity to methacholine and an increased mortality when exposed to bacterial endotoxin. An aberrant melanogenesis and proliferative ab...

  14. Endotoxina e câncer Endotoxin and cancer

    Directory of Open Access Journals (Sweden)

    Jessica I. Lundin

    2010-09-01

    Full Text Available A exposição à endotoxina, componente de paredes celulares bacterianas gram-negativas, é muito comum em plantas industriais e no meio ambiente. Ambientes de alta exposição incluem fazendas de criação de animais, instalações têxteis de algodão e moinhos. Neste artigo, revemos estudos experimentais, epidemiológicos e ensaios clínicos sobre a hipótese de que a endotoxina previne o câncer. Desde os anos 70, estudos epidemiológicos em têxteis de algodão e outros grupos ocupacionais expostos à endotoxina demonstram redução no risco de câncer de pulmão. Pesquisa experimental de toxicologia animal e ensaios terapêuticos limitados em pacientes com câncer dão suporte para um potencial anticarcinogênico. Os mecanismos biológicos anticarcinogênicos de base ainda não são completamente compreendidos, mas acredita-se que incluem recrutamento e ativação de células imunológicas e mediadores pró-inflamatórios (ex.: fator de necrose tumoral α e interleucina-1 e - 6. Devido ao estágio atual de conhecimento, seria prematuro recomendar a endotoxina como agente quimiopreventivo. Porém, pesquisas epidemiológicas e experimentais que esclareçam relações de dosagem-efeito e exposição-relações temporais podem trazer benefícios para a saúde pública e a biomedicina básica.Exposure to endotoxin, a component of gram-negative bacterial cell walls, is widespread in many industrial settings and in the ambient environment. Heavy-exposure environments include livestock farms, cotton textile facilities, and saw mills. In this article, we review epidemiologic, clinical trial, and experimental studies pertinent to the hypothesis that endotoxin prevents cancer. Since the 1970s, epidemiologic studies of cotton textile and other endotoxin-exposed occupational groups have consistently demonstrated reduced lung cancer risks. Experimental animal toxicology research and some limited therapeutic trials in cancer patients offer additional

  15. Polaprezinc Protects Mice against Endotoxin Shock.

    Science.gov (United States)

    Ohata, Shuzo; Moriyama, Chihiro; Yamashita, Atsushi; Nishida, Tadashi; Kusumoto, Chiaki; Mochida, Shinsuke; Minami, Yukari; Nakada, Junya; Shomori, Kohei; Inagaki, Yoshimi; Ohta, Yoshiji; Matsura, Tatsuya

    2010-05-01

    Polaprezinc (PZ), a chelate compound consisting of zinc and l-carnosine (Car), is an anti-ulcer drug developed in Japan. In the present study, we investigated whether PZ suppresses mortality, pulmonary inflammation, and plasma nitric oxide (NO) and tumor necrosis factor (TNF)-alpha levels in endotoxin shock mice after peritoneal injection of lipopolysaccharide (LPS), and how PZ protects against LPS-induced endotoxin shock. PZ pretreatment inhibited the decrease in the survival rate of mice after LPS injection. PZ inhibited the increases in plasma NO as well as TNF-alpha after LPS. Compatibly, PZ suppressed LPS-induced inducible NO synthase mRNA transcription in the mouse lungs. PZ also improved LPS-induced lung injury. However, PZ did not enhance the induction of heat shock protein (HSP) 70 in the mouse lungs after LPS. Pretreatment of RAW264 cells with PZ suppressed the production of NO and TNF-alpha after LPS addition. This inhibition likely resulted from the inhibitory effect of PZ on LPS-mediated nuclear factor-kappaB (NF-kappaB) activation. Zinc sulfate, but not Car, suppressed NO production after LPS. These results indicate that PZ, in particular its zinc subcomponent, inhibits LPS-induced endotoxin shock via the inhibition of NF-kappaB activation and subsequent induction of proinflammatory products such as NO and TNF-alpha, but not HSP induction. PMID:20490319

  16. Endotoxin and cancer chemo-prevention.

    Science.gov (United States)

    Mastrangelo, Giuseppe; Fadda, Emanuela; Cegolon, Luca

    2013-10-01

    Reduced rates of lung cancer have been observed in several occupational groups exposed to high levels of organic dusts contaminated by endotoxin. The underlying anti-neoplastic mechanism of endotoxin may be an increased secretion of endogenous anti-neoplastic mediators and activation of the toll-like receptors (TLR). A detoxified endotoxin derivative, Monophosphoryl Lipid A (MPL(®)) is marketed in Europe since 1999 as part of the adjuvant systems in allergy vaccines for treatment of allergic rhino-conjunctivitis and allergic asthma. Over 200,000 patients have used them to date (nearly 70% in Germany). Since detailed exposure (MPL(®) dose and timing of administration) and individual data are potentially available, an observational follow-up study could be conducted in Germany to investigate the protective effect of MPL(®) against cancer, comparing cancer incidence in two groups of patients with allergic rhinitis: those treated with allergoids plus MPL(®) and those treated with a vaccine including the same allergoids but not MPL(®). The protective effect of MPL(®) could be quantified in ever and never smokers. If this proposed observational study provides evidence of protective effects, MPL(®) could be immediately used as a chemo-preventive agent since it is already in use as adjuvant in human vaccines against cancer.

  17. Effects of grain, fructose, and histidine feeding on endotoxin and oxidative stress measures in dairy heifers.

    Science.gov (United States)

    Golder, H M; Lean, I J; Rabiee, A R; King, R; Celi, P

    2013-01-01

    Ruminal endotoxin and plasma oxidative stress biomarker concentrations were studied in dairy heifers challenged with grain, fructose, and histidine in a partial factorial study. Holstein-Friesian heifers [n=30; average body weight (BW) of 359.3±47.3 kg] were randomly allocated to 5 treatment groups: (1) control (no grain); (2) grain [crushed triticale at 1.2% of BW dry matter intake (DMI)]; (3) grain (0.8% of BW DMI) + fructose (0.4% of BW DMI); (4) grain (1.2% of BW DMI) + histidine (6g/head); and (5) grain (0.8% of BW DMI) + fructose (0.4% of BW DMI) + histidine (6 g/head). Rumen samples were collected by stomach tube 5, 65, 115, 165, and 215 min after diet consumption and blood samples at 5 and 215 min after consumption. Rumen fluid was analyzed for endotoxin concentrations. Plasma was analyzed for concentrations of the following oxidative stress biomarkers: reactive oxygen metabolites (dROM), biological antioxidant potential (BAP), advanced oxidation protein products, and ceruloplasmin, and activity of glutathione peroxidase. Dietary treatment had no effect on concentrations of endotoxin or oxidative stress biomarkers. We observed no interactions of treatment by time. Ruminal concentrations of endotoxin decreased during the sampling period from 1.12×10(5) ± 0.06 to 0.92×10(5) endotoxin units/mL ± 0.05 (5 and 215 min after diet consumption, respectively). Concentrations of dROM and the oxidative stress index (dROM/BAP × 100) increased over the sampling period, from 108.7 to 123.5 Carratelli units (Carr U), and from 4.1 to 4.8, respectively. Ceruloplasmin concentrations markedly declined 5 min after the consumption of diets, from 190 to 90 mg/L over the 215-min sampling period. Overall, a single feeding challenge for dairy cattle with grain, fructose, and histidine, and combinations thereof, may not be sufficient to induce marked changes in endotoxin or oxidative stress biomarker concentrations.

  18. Effects of grain, fructose, and histidine feeding on endotoxin and oxidative stress measures in dairy heifers.

    Science.gov (United States)

    Golder, H M; Lean, I J; Rabiee, A R; King, R; Celi, P

    2013-01-01

    Ruminal endotoxin and plasma oxidative stress biomarker concentrations were studied in dairy heifers challenged with grain, fructose, and histidine in a partial factorial study. Holstein-Friesian heifers [n=30; average body weight (BW) of 359.3±47.3 kg] were randomly allocated to 5 treatment groups: (1) control (no grain); (2) grain [crushed triticale at 1.2% of BW dry matter intake (DMI)]; (3) grain (0.8% of BW DMI) + fructose (0.4% of BW DMI); (4) grain (1.2% of BW DMI) + histidine (6g/head); and (5) grain (0.8% of BW DMI) + fructose (0.4% of BW DMI) + histidine (6 g/head). Rumen samples were collected by stomach tube 5, 65, 115, 165, and 215 min after diet consumption and blood samples at 5 and 215 min after consumption. Rumen fluid was analyzed for endotoxin concentrations. Plasma was analyzed for concentrations of the following oxidative stress biomarkers: reactive oxygen metabolites (dROM), biological antioxidant potential (BAP), advanced oxidation protein products, and ceruloplasmin, and activity of glutathione peroxidase. Dietary treatment had no effect on concentrations of endotoxin or oxidative stress biomarkers. We observed no interactions of treatment by time. Ruminal concentrations of endotoxin decreased during the sampling period from 1.12×10(5) ± 0.06 to 0.92×10(5) endotoxin units/mL ± 0.05 (5 and 215 min after diet consumption, respectively). Concentrations of dROM and the oxidative stress index (dROM/BAP × 100) increased over the sampling period, from 108.7 to 123.5 Carratelli units (Carr U), and from 4.1 to 4.8, respectively. Ceruloplasmin concentrations markedly declined 5 min after the consumption of diets, from 190 to 90 mg/L over the 215-min sampling period. Overall, a single feeding challenge for dairy cattle with grain, fructose, and histidine, and combinations thereof, may not be sufficient to induce marked changes in endotoxin or oxidative stress biomarker concentrations. PMID:24119801

  19. A true theranostic approach to medicine: towards tandem sensor detection and removal of endotoxin in blood.

    Science.gov (United States)

    Thompson, Michael; Blaszykowski, Christophe; Sheikh, Sonia; Romaschin, Alexander

    2015-05-15

    Sepsis is one of the leading causes of death around the world. The condition occurs when a local infection overcomes the host natural defense mechanism and suddenly spreads into the circulatory system, triggering a vigorous, self-injurious inflammatory host response. The pathogenesis of sepsis is relatively well known, one of the most potent immuno-activator being bacterial lipopolysaccharide (LPS) - also known as 'endotoxin'. Tests exist to detect endotoxin in bodily fluids, but are expensive, not necessarily user-friendly and require reporter molecules. In addition, the situation for safe and effective anti-endotoxin therapy is problematical. At the present time, endotoxin removal through cartridge hemoperfusion is one of the better alternatives to combat sepsis. The capability to both measure endotoxemia levels and offer an adapted response treatment in a timely manner is crucial for better management and improved prognosis, but is currently unavailable. In this context, we describe herein preliminary research towards the development of an alternative LPS biosensor and an innovative LPS neutralization cartridge to be eventually combined in an all-integrated configuration for the theranostic, personalized treatment of blood endotoxemia/sepsis. LPS detection is performed in a real-time and label-free manner in full human blood plasma, using ultra-high frequency acoustic wave sensing in combination with ultrathin, oligoethylene glycol-based mixed surface chemistry imposed on piezoelectric quartz discs. Biosensing platforms are functionalized with polymyxin B (PMB), a cyclic peptide antibiotic with high affinity for LPS. Analogous surface modification is used on glass beads for the therapeutic cartridge component of the combined strategy. Incubation of LPS-spiked whole blood with PMB-bead chemistry resulted in a significant decrease in the production of pro-inflammatory TNF-α cytokine. LPS neutralization is discussed in relation to the perturbation of its

  20. Endotoxin elicits ambivalent social behaviors

    OpenAIRE

    Yee, Jason R.; Prendergast, Brian J.

    2011-01-01

    The acute phase response to infection is reliably accompanied by decreases in social investigation; however, social behavior is commonly assayed in inescapable environments using unfamiliar social stimuli. In this experiment, male Wistar rats were raised from weaning with 2 familiar, same-sex conspecifics. In adulthood, rats were implanted with radiotelemetry devices that permitted localization in space, and were challenged with LPS treatments (150 mg/kg, i.p.) in a novel, semi-natural arena ...

  1. 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...

  2. 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 concentr

  3. 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...

  4. Determination of endotoxin through an aptamer-based impedance biosensor.

    Science.gov (United States)

    Su, Wenqiong; Lin, Meng; Lee, Hyuck; Cho, MiSuk; Choe, Woo-Seok; Lee, Youngkwan

    2012-02-15

    Lipopolysaccharide (LPS) often referred to endotoxin is an undesirable impurity frequently entrained with various recombinant protein therapeutics and plasmid DNA (pDNA) vaccines of bacterial origin. The inherent toxicities (e.g. fever, hypotension, shock and death) of LPS render its early and sensitive detection essential for several biological assays and/or parenteral administrations of biotherapeutics. In this study, an electrochemical biosensor using an LPS specific single stranded DNA (ssDNA) aptamer as a probe was developed. Amine-terminated aptamer exhibiting high affinity (K(d)=11.9 nM) to LPS was immobilized on a gold electrode using 3-mercaptopropionic acid (MPA) as a linker. Each step of the modification process was characterized by cyclic voltammetry (CV) and electrochemical impendence spectroscopy (EIS). A good linear relationship of the changes in the charge-transfer resistance (ΔR(et)) and the logarithmic value of LPS concentration was demonstrated in a broad dynamic detection range of 0.001-1 ng/ml. Furthermore, the aptasensor showed a high selectivity to LPS despite the presence of pDNA, RNA and bovine serum albumin (BSA) and could be regenerated in low pH condition, offering a promising option for detecting LPS often present in a complex milieu. PMID:22182428

  5. 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.

  6. Co-expression of heat shock protein (HSP) 40 and HSP70 in Pinctada martensii response to thermal, low salinity and bacterial challenges.

    Science.gov (United States)

    Li, Jun; Zhang, Yuehuan; Liu, Ying; Zhang, Yang; Xiao, Shu; Yu, Ziniu

    2016-01-01

    Heat shock protein (HSP) 40 proteins are a family of molecular chaperones that bind to HSP70 through their J-domain and regulate the function of HSP70 by stimulating its adenosine triphosphatase activity. In the present study, a HSP40 homolog named PmHSP40 was cloned from the hemocytes of pearl oyster Pinctada martensii using EST and rapid amplification of cDNA ends (RACE) techniques. The full-length cDNA of PmHSP40 was 1251 bp in length, which included a 5' untranslated region (UTR) of 75 bp, an open reading frame (ORF) of a 663 bp, and a 3' UTR of 513 bp. The deduced amino acid sequence of PmHSP40 contains a J domain in the N-terminus. In response to thermal and low salinity stress challenges, the expression of PmHSP40 in hemocytes and the gill were inducible in a time-dependent manner. After bacterial challenge, PmHSP40 transcripts in hemocytes increased and peaked at 6 h post injection. In the gill, PmHSP40 expression increased, similar to expression in hemocytes; however, transcript expression of PmHSP40 was significantly up-regulated at 12 h post injection. Furthermore, the transcripts of PmHSP70 showed similar kinetics as that of PmHSP40, with highest induction during thermal, low salinity stress and bacterial challenges. Altogether these results demonstrate that PmHSP40 is an inducible protein under thermal, low salinity and bacterial challenges, suggesting its involvement in both environmental and biological stresses, and in the innate immunity of the pearl oyster. PMID:26679110

  7. Development of an anti-endotoxin vaccine for sepsis.

    Science.gov (United States)

    Cross, Alan S

    2010-01-01

    Gram-negative bacterial lipopolysaccharide (LPS, endotoxin) is an important initiator of sepsis, a clinical syndrome that is a leading cause of death in intensive care units. Vaccines directed against core LPS structures that are widely conserved among Gram-negative bacteria (GNB) have been developed for the treatment and/or prevention of sepsis. Killed whole bacterial vaccines (E. coli O111:B4, J5 [Rc chemotype] mutant and S. minnesota, Re chemotype) protected mice against experimental sepsis. Human J5 immune antisera reduced the mortality from GNB sepsis in a large controlled clinical trial; however, subsequent clinical studies with antiendotoxin antibodies did not demonstrate protective efficacy in sepsis. Multiple clinical studies have since demonstrated a correlation between the level of circulating antibodies to LPS core and morbidity and mortality in different clinical settings. We therefore developed a subunit vaccine by combining detoxified J5 LPS (J5 dLPS) with the outer membrane protein (OMP) from group B N. meningitidis. This vaccine was highly efficacious in experimental models of sepsis and progressed to phase 1 clinical trial. While well-tolerated, this vaccine induced only 3-4-fold increases in anti-J5 dLPS antibody. Addition of the TLR9 agonist, oligodeoxynucleotide with a CpG motif, as adjuvant to the vaccine increased antibody levels in mice and the vaccine/CpG combination will progress to phase 1 human study. Additional vaccines in which the core glycolipid was either conjugated to carrier protein or incorporated into liposomes have been developed, but have not progressed to clinical trial. Should an antiendotoxin vaccine become available, a new immunization strategy directed towards distinct populations at risk will be required. PMID:20593272

  8. Sleep deprivation attenuates endotoxin-induced cytokine gene expression independent of day length and circulating cortisol in male Siberian hamsters (Phodopus sungorus).

    Science.gov (United States)

    Ashley, Noah T; Walton, James C; Haim, Achikam; Zhang, Ning; Prince, Laura A; Fruchey, Allison M; Lieberman, Rebecca A; Weil, Zachary M; Magalang, Ulysses J; Nelson, Randy J

    2013-07-15

    Sleep is restorative, whereas reduced sleep leads to negative health outcomes, such as increased susceptibility to disease. Sleep deprivation tends to attenuate inflammatory responses triggered by infection or exposure to endotoxin, such as bacterial lipopolysaccharide (LPS). Previous studies have demonstrated that Siberian hamsters (Phodopus sungorus), photoperiodic rodents, attenuate LPS-induced fever, sickness behavior and upstream pro-inflammatory gene expression when adapted to short day lengths. Here, we tested whether manipulation of photoperiod alters the suppressive effects of sleep deprivation upon cytokine gene expression after LPS challenge. Male Siberian hamsters were adapted to long (16 h:8 h light:dark) or short (8 h:16 h light:dark) photoperiods for >10 weeks, and were deprived of sleep for 24 h using the multiple platform method or remained in their home cage. Hamsters received an intraperitoneal injection of LPS or saline (control) 18 h after starting the protocol, and were killed 6 h later. LPS increased liver and hypothalamic interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF) gene expression compared with vehicle. Among LPS-challenged hamsters, sleep deprivation reduced IL-1 mRNA levels in liver and hypothalamus, but not TNF. IL-1 attenuation was independent of circulating baseline cortisol, which did not increase after sleep deprivation. Conversely, photoperiod altered baseline cortisol, but not pro-inflammatory gene expression in sleep-deprived hamsters. These results suggest that neither photoperiod nor glucocorticoids influence the suppressive effect of sleep deprivation upon LPS-induced inflammation.

  9. Intestinal radiation syndrome: sepsis and endotoxin

    International Nuclear Information System (INIS)

    Rats were whole-body irradiated with 8-MeV cyclotron-produced neutrons and 137Cs γ 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 γ-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

  10. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  12. Endotoxin Elimination in Patients with Septic Shock: An Observation Study.

    Science.gov (United States)

    Adamik, Barbara; Zielinski, Stanislaw; Smiechowicz, Jakub; Kübler, Andrzej

    2015-12-01

    To evaluate the effectiveness of endotoxin elimination with an adsorption column in patients with septic shock and endotoxemia. The elimination therapy was guided by a new bedside method of measuring endotoxin activity (EA). Intensive care unit (ICU) patients with septic shock and suspected Gram-negative infection were consecutively added to the study group within the first 24 h. Endotoxin elimination was performed using hemoperfusion with the Alteco LPS Adsorber. The primary endpoint was improvement in organ function within the first 24 h of treatment. A secondary objective was to assess the usefulness of a new method of measuring EA to help guide endotoxin elimination therapy. Out of 64 patients 18 had a high baseline EA [0.70 EA units (0.66-0.77)]. Those patients had endotoxin elimination treatment in addition to conventional medical therapy. At 24 h after endotoxin elimination, the EA had decreased to 0.56 EA units (0.43-0.77), (p = 0.005); MAP increased from 69 (62-80) to 80 mm Hg (68-88), (p = 0.002), and noradrenaline use decreased from 0.28 (0.15-0.80) to 0.1 μg/kg/min (0.00-0.70) at the same time (p = 0.04). The SOFA score had decreased from 11 (9-15) to 9 (7-14) points 24 h after endotoxin elimination (p = 0.01) with a median delta SOFA -2 points. Endotoxin elimination did not have a significant effect on the ICU length of stay or ICU mortality. Effective endotoxin elimination resulted in a significant improvement in hemodynamic parameters and of organ function. The application of the EA assay was useful for the bedside monitoring of endotoxemia in critically ill ICU patients.

  13. Elevated endotoxin levels in non-alcoholic fatty liver disease

    Directory of Open Access Journals (Sweden)

    Kumar Sudhesh

    2010-03-01

    Full Text Available Abstract Background Emerging data indicate that gut-derived endotoxin may contribute to low-grade systemic inflammation in insulin resistant states. This study aimed to examine the importance of serum endotoxin and inflammatory markers in non-alcoholic fatty liver disease (NAFLD patients, with and without type 2 diabetes mellitus (T2DM, and to explore the effect of treatment with a lipase inhibitor, Orlistat, on their inflammatory status. Methods Fasted serum from 155 patients with biopsy proven NAFLD and 23 control subjects were analysed for endotoxin, soluble CD14 (sCD14, soluble tumour necrosis factor receptor II (sTNFRII and various metabolic parameters. A subgroup of NAFLD patients were re-assessed 6 and 12 months after treatment with diet alone (n = 6 or diet plus Orlistat (n = 8. Results Endotoxin levels were significantly higher in patients with NAFLD compared with controls (NAFLD: 10.6(7.8, 14.8 EU/mL; controls: 3.9(3.2, 5.2 EU/mL, p Sub-cohort treatment with Orlistat in patients with NAFLD showed significant decreases in ALT (p = 0.006, weight (p = 0.005 and endotoxin (p = 0.004 compared with the NAFLD, non-Orlistat treated control cohort at 6 and 12 months post therapy, respectively. Conclusions Endotoxin levels were considerably increased in NAFLD patients, with marked increases noted in early stage fibrosis compared with controls. These results suggest elevated endotoxin may serve as an early indicator of potential liver damage, perhaps negating the need for invasive liver biopsy. As endotoxin may promote insulin resistance and inflammation, interventions aimed at reducing endotoxin levels in NAFLD patients may prove beneficial in reducing inflammatory burden.

  14. A matrix of cholesterol crystals, but not cholesterol alone, primes human monocytes/macrophages for excessive endotoxin-induced production of tumor necrosis factor-alpha. Role in atherosclerotic inflammation?

    DEFF Research Database (Denmark)

    Bendtzen, Klaus; Christensen, Ole; Nielsen, Claus Henrik;

    2014-01-01

    When exposed to small amounts of bacterial endotoxin, matrices of cholesterol crystals, but not cholesterol itself, primed human monocytes/macrophages to a highly augmented (>10-fold) production of inflammatory tumor necrosis factor-α. Priming also sensitized the cells, as 10- to 100-fold lower...... levels of endotoxin were needed for TNF-α production equivalent to that of unprimed cells. The pro-inflammatory effect was selective as endotoxin-induced production of other pro-inflammatory cytokines was unaffected while production of anti-inflammatory interleukin-10 was diminished. These findings...... suggest that cholesterol matrix formation may play a pathogenic role in atherosclerotic inflammation, and they indicate a mechanism by which bacteria and/or bacterial products may play a role in processes leading to arteriosclerosis....

  15. 动态浊度法测定红花注射液中细菌内毒素的含量%Content Determination of BacteriaI Endotoxin in Honghua Injection by Ki-netic Turbidimetric LimuIus Method

    Institute of Scientific and Technical Information of China (English)

    李长根

    2015-01-01

    目的:建立动态浊度法测定红花注射液中细菌内毒素的含量。方法采用动态浊度法,通过测定供试液中外加内毒素的回收率进行干扰试验,确定样品无干扰浓度,并定量样品中细菌内毒素。结果内毒素检查质量浓度线性范围为0.03125~2.0EU・ mL -1(r=0.9966),样品在稀释10倍以上对试验无干扰,细菌内毒素回收率在50%~200%范围内,样品中内毒素可定量测定。结论动态浊度法可用于红花注射液中细菌内毒素的含量检测。%OBJECTIVE To establish a method for determining the content of bacterial endotoxin in Honghua Injection.METHODS Using Kinetic Turbidimetric Limulus Method, sample was confirmed without interference, and bacterial endotoxin in the sample was quantitated by interference test with measuring the endotoxin additional in test liquid.RESULTS The linear range of bacterial endotoxin detection concentration was 0.31~2.0 EU・ mL-1 ( r=0.9981 );the sample under Diluted 10 times had no interference to test.The recovery rate of bacterial endotoxin was within the range of 50%~200%.The bacterial endotoxin content in the sample could be quantitatively deter-mined.CONCLUSION The kinetic turbidimetric limulus method can be used to the quantitative determination of bacterial endotoxin in Honghua Injection.

  16. Allergen and endotoxin exposure in a companion animal hospital.

    OpenAIRE

    Samadi, S.; Heederik, D.J.J.; Krop, E. J. M.; Jamshidifard, A.R.; Willemse, T; Wouters, I.M.

    2010-01-01

    BACKGROUND: Exposure to allergens, both in general and occupational environments, is known to result in sensitisation and exacerbation of allergic diseases, while endotoxin exposure might protect against allergic diseases. This may be important for veterinarians and co-workers. However, exposure levels are mostly unknown. OBJECTIVE: We investigated the allergen and endotoxin exposure levels of veterinary medicine students and workers in a companion animal hospital. METHODS: Airborne and surfa...

  17. Endotoxin in Size-Separated Metal Working Fluid Aerosol Particles.

    Science.gov (United States)

    Dahlman-Höglund, Anna; Lindgren, Åsa; Mattsby-Baltzer, Inger

    2016-08-01

    Patients with airway symptoms working in metal working industries are increasing, despite efforts to improve the environmental air surrounding the machines. Our aim was to analyse the amount of endotoxin in size-separated airborne particles of metal working fluid (MWF) aerosol, by using the personal sampler Sioutas cascade impactor, to compare filter types, and to compare the concentration of airborne endotoxin to that of the corresponding MWFs. In a pilot field study, aerosols were collected in two separate machine halls on totally 10 occasions, using glass fibre and polytetrafluoroethylene (PTFE) filters in parallel at each station. Airborne endotoxin was distributed over all size fractions. While a major part was found in the largest size fraction (72%, 2.5-10 µm), up to 8% of the airborne endotoxin was detected in the smallest size fraction (efficiency of the filter types, a significantly higher median endotoxin level was found with glass fibres filters collecting the largest particle-size fraction (1.2-fold) and with PTFE filters collecting the smallest ones (5-fold). The levels of endotoxin in the size-separated airborne particle fractions correlated to those of the MWFs supporting the aerosol-generating machines. Our study indicates that a significant part of inhalable aerosols of MWFs consists of endotoxin-containing particles below the size of intact bacteria, and thus small enough to readily reach the deepest part of the lung. Combined with other chemical irritants of the MWF, exposure to MWF aerosols containing endotoxin pose a risk to respiratory health problems. PMID:27268595

  18. Evolution of endotoxin induced acute lung injury in the rat.

    OpenAIRE

    Domenici-Lombardo, L.; C. Adembri; Consalvo, M.; Forzini, R.; Meucci, M.; Romagnoli, P; Novelli, G.P.

    1995-01-01

    To clarify the evolution of acute lung injury induced by endotoxin, the progression of lung damage in 26 rats submitted to intratracheal instillation of 5 mg/kg body weight endotoxin was examined by blood gas analysis, computerized tomography, light and electron microscopy. Hypoxaemia, hypercapnia, acidosis and inhomogeneous bilateral infiltrates developed gradually within 48 hours. Monocytes appeared within blood capillaries and the instertitium by 12 hours after treatment, then migrated int...

  19. Ultrasensitive detection of endotoxins using computationally designed nanoMIPs.

    Science.gov (United States)

    Altintas, Zeynep; Abdin, Mohammed J; Tothill, Alexander M; Karim, Kal; Tothill, Ibtisam E

    2016-09-01

    Novel molecularly imprinted polymer nanoparticles (nanoMIPs) were designed for endotoxin from Escherichia coli 0111:B4, using computational modeling. The screening process based on binding energy between endotoxin and each monomer was performed with 21 commonly used monomers, resulting in the selection of itaconic acid, methacrylic acid and acrylamide as functional monomers due to their strong binding interaction with the endotoxin template. The nanoMIPs were successfully synthesized with functional groups on the outer surface to aid in the immobilization onto sensor surface. The solid phase photopolymerization approach used for the synthesis of nanoMIPs ranging from 200 to 235 nm in diameter. The limit of detection and KD were significantly improved when endotoxin samples were prepared using a novel triethylamine method. This improved the efficiency of gold nanoparticle functionalization by targeting the subunits of the endotoxin. Compared to the vancomycin MIP control, the endotoxin MIPs displayed outstanding affinity and selectivity towards the endotoxin with KD values in the range of 4.4-5.3 × 10(-10) M, with limits of detection of 0.44 ± 0.02 ng mL(-1) as determined by surface plasmon resonance (SPR) sensor when itaconic acid was used as the functional monomer. The MIP surface can be regenerated >30 times without significant loss of binding activity making this approach highly cost effective for expensive analyte templates. The combination of molecular modeling and solid phase synthesis enabled the successful synthesis of nanoMIPs capable of recognition and ultrasensitive detection of endotoxins using the highly sensitive SPR biosensor with triethylamine method. PMID:27543033

  20. Characterization and expression analysis of a peptidoglycan recognition protein gene, SmPGRP2 in mucosal tissues of turbot (Scophthalmus maximus L.) following bacterial challenge.

    Science.gov (United States)

    Zhang, Linan; Gao, Chengbin; Liu, Fengqiao; Song, Lin; Su, Baofeng; Li, Chao

    2016-09-01

    Peptidoglycan recognition receptor proteins (PGRPs), a group of pattern recognition receptors (PRRs), can recognize peptidoglycan (PGN) of the bacteria cell wall and play an important role in host immune defense against pathogen infection. They are highly structurally conserved through evolution, but with different function in innate immunity between invertebrates and vertebrates. In teleost fish, several PGRPs have been characterized recently. They have both amidase activity and bactericidal activity and are involved in indirectly killing bacteria and regulating multiple signaling pathways. However, the knowledge of PGRPs in mucosal immunity of teleost fish is still limited. In this study, we identified a PGRPs gene (SmPGRP2) of turbot and investigated its expression patterns in mucosal tissues after challenge with Gram-positive bacteria Streptococcus iniae and Gram-negative bacteria Vibrio anguillarum. Phylogenetic analysis showed the strongest relationship of turbot PGRP to halibut, which was consistent with their phylogenetic relationships. In addition, SmPGRP2 was ubiquitously expressed in turbot tissues, and constitutive expression levels were higher in classical immune tissues (including liver, spleen, and head-kidney) than mucosal tissues (intestine, gill and skin). After bacterial challenge, the expression of SmPGRP2 was induced and showed a general trend of up-regulation in mucosal tissues, except in intestine following V. anguillarum infection. These different expression patterns varied depending on both pathogen and tissue type, suggesting its distinct roles in the host immune response to bacterial pathogen. PMID:27461422

  1. Facile biofunctionalization of silver nanoparticles for enhanced antibacterial properties, endotoxin removal, and biofilm control

    Directory of Open Access Journals (Sweden)

    Lambadi PR

    2015-03-01

    Full Text Available Paramesh Ramulu Lambadi,1,* Tarun Kumar Sharma,1,* Piyush Kumar,1 Priyanka Vasnani,2 Sitaramanjaneya Mouli Thalluri,2 Neha Bisht,1 Ranjana Pathania,1,2 Naveen Kumar Navani1,21Department of Biotechnology, 2Centre of Nanotechnology, Indian Institute of Technology, Roorkee, Uttarakhand, India*These authors contributed equally to this workAbstract: Infectious diseases cause a huge burden on healthcare systems worldwide. Pathogenic bacteria establish infection by developing antibiotic resistance and modulating the host’s immune system, whereas opportunistic pathogens like Pseudomonas aeruginosa adapt to adverse conditions owing to their ability to form biofilms. In the present study, silver nanoparticles were biofunctionalized with polymyxin B, an antibacterial peptide using a facile method. The biofunctionalized nanoparticles (polymyxin B-capped silver nanoparticles, PBSNPs were assessed for antibacterial activity against multiple drug-resistant clinical strain Vibrio fluvialis and nosocomial pathogen P. aeruginosa. The results of antibacterial assay revealed that PBSNPs had an approximately 3-fold higher effect than the citrate-capped nanoparticles (CSNPs. Morphological damage to the cell membrane was followed by scanning electron microscopy, testifying PBSNPs to be more potent in controlling the bacterial growth as compared with CSNPs. The bactericidal effect of PBSNPs was further confirmed by Live/Dead staining assays. Apart from the antibacterial activity, the biofunctionalized nanoparticles were found to resist biofilm formation. Electroplating of PBSNPs onto stainless steel surgical blades retained the antibacterial activity against P. aeruginosa. Further, the affinity of polymyxin for endotoxin was exploited for its removal using PBSNPs. It was found that the prepared nanoparticles removed 97% of the endotoxin from the solution. Such multifarious uses of metal nanoparticles are an attractive means of enhancing the potency of antimicrobial

  2. Pavlovian conditioning of endotoxin-tolerance in rats.

    Science.gov (United States)

    Oberbeck, Reiner; Kromm, Alexander; Exton, Michael S; Schade, Ullrich; Schedlowski, Manfred

    2003-02-01

    The most fascinating example of the bi-directional interaction between the central nervous system (CNS) and immune system is the behavioral conditioning of immune functions. We therefore investigated the behavioral conditioning of lipopolysaccharide (LPS)-induced endotoxin tolerance using the taste aversion paradigm. The conditioned stimulus (CS) saccharin was paired with the unconditioned stimulus (UCS) LPS over a five (CONDl) or four (COND2) days learning trial. Controls received drinking water with (SHAM) or without (UNT) LPS. Endotoxin tolerance was tested by determination of LPS-induced tumor necrosis factor (TNF)-alpha release. After the avoidance of the induced endotoxin-tolerance the CS saccharin was re-presented in all experimental groups. A the end of the re-exposure period a complete endotoxin tolerance was noticed in the CONDl- and COND2-group. In contrast, no effect of saccharin administration was observed in the SHAM- or UNT-group. Our data demonstrate for the first time the behavioral conditioning of endotoxin tolerance. Furthermore, these results contribute new aspects to the mechanisms underlying the development and modulation of endotoxin tolerance.

  3. Inflammatory responses in epithelia: endotoxin-induced IL-6 secretion and iNOS/NO production are differentially regulated in mouse mammary epithelial cells

    OpenAIRE

    2010-01-01

    Background IL-6 is a pro-inflammatory cytokine that signals via binding to a soluble or membrane bound receptor, while nitric oxide (NO), an oxidative stress molecule, diffuses through the cell membrane without a receptor. Both mediators signal through different mechanisms, yet they are dependent on NFκB. We proposed that both mediators are co-induced and co-regulated in inflamed mammary epithelial cells. Methods SCp2 mammary epithelial cells were treated with bacterial endotoxin (ET) for dif...

  4. The 3-hydroxy fatty acids as biomarkers for quantification and characterization of endotoxins and Gram-negative bacteria in atmospheric aerosols in Hong Kong

    Science.gov (United States)

    Lee, Alex K. Y.; Chan, Chak K.; Fang, Ming; Lau, Arthur P. S.

    Endotoxins from Gram-negative bacteria have received much attention because they could elicit strong pro-inflammatory responses in the human respiratory tract. In this study, 3-hydroxy fatty acids (3-OH FAs) with carbon chain lengths from 10 to 18 (C10-C18) were employed as biomarkers to quantify and characterize the endotoxins and Gram-negative bacterial community in atmospheric aerosols. Gas chromatography-mass spectrometry (GC-MS) was utilized for quantification of this biomarker in fine (PM 2.5) and coarse (PM 2.5-10) particulates collected by high volume samplers simultaneously at a rural and an urban site in Hong Kong. The geometric mean concentrations of the endotoxins were 5.5 and 1.35 ng m -3 in fine and coarse particulates at the rural site, respectively. At the urban site, the corresponding concentrations were 9.4 and 2.80 ng m -3 in fine and coarse particulates, respectively. It is found that 70-80% of the total endotoxins are associated with the fine particulates. Significant higher endotoxin levels at the urban site were observed throughout the 8-month study period. This could possibly relate to the heavier human activities in the urban areas. The distribution patterns of the 3-OH FAs with respect to carbon number are similar between the rural and urban sites regardless of particle sizes. The C10 and C16 were predominant and accounted for about 40-50% of the total 3-OH FAs. Furthermore, the odd carbon chain length 3-OH FAs constituted a non-negligible fraction (15-25%) of the total 3-OH FAs. The biologically active endotoxins estimated as the sum of C12 and C14 portions in this study ranged from 0.6-3.7 and 1.9-4.8 ng m -3 at the rural and urban sites, respectively. Applying the biomarker-to-microbial mass conversion factors, the dry mass loading of the Gram-negative bacteria are in the order of 10-10 2 ng m -3 in atmospheric aerosol. This study also demonstrates that the biomarker (3-OH FAs) approach yields much more quantitative information such as

  5. Experimental study on the role of endotoxin in the development of hepatopulmonary syndrome

    Institute of Scientific and Technical Information of China (English)

    Hui-Ying Zhang; De-Wu Han; Xin-Guo Wang; Yuan-Chang Zhao; Xin Zhou; Hai-Zhen Zhao

    2005-01-01

    AIM: To evaluate the role of intestinal endotoxemia in the genesis of hepatopulmonary syndrome.METHODS: A rat model of cirrhosis was prepared with the method of compound factors. At the end of the eighth week, rats with cirrhosis were treated with 300 μg LPS/100 g body weight, and 1 g/rat of glycine about four h prior to LPS. After three h of LPS treatment, blood and tissues were collected for various measurements. Kupffer cells were isolated from male Wistar rats and cultured, and divided into five groups. Supernatant was harvested at 3 h after treatment with LPS for measurement of tumor necrosis factor-alpha (TNF-α).RESULTS: Our results showed that in rats with cirrhosis, zslowed and deepened breath with occasional pause was.PaO2, PaCO2 and standard bicarbonate (SB) in arterial blood were decreased. Arterial O2 and actual bicarbonate (AB) were markedly decreased. There was a close correlation between decreased O2 and endotoxin. Metabolic acidosis accompanying respiratory alkalosis was the primary type of acid-base imbalance. The alveolar-arterial oxygen gradient was sharply widened. Massive accumulation of giant macrophages in the alveolar spaces and its wall and widened alveolar wall architecture were observed. The number of bacterial translocations in mesenteric lymph nodes increased. The ratio of TC99M-MAA brain-over-lung radioactivity rose. Endotoxin, and TNF-α, endothelin-1 (ET-1), nitric oxide (NO) in plasma and ET-1, carbon monoxide (CO) in lung homogenates increased. After administration of a given dosage of LPS in rats with cirrhosis, various pathological parameters worsened.Plasma level of endotoxin was related to TNF-α, ET-1, NO in plasma and ET-1, NO, CO in lung homogenates. TNF-α level was related to ET-1 and NO in plasma and lung homogenates and CO in lung homogenate as well. The level of TNF-α increased after infusion of LPS into culture supernatant of Kupffer cells in vitro. However, TNF-α significantly decreased after pretreatment with

  6. Endotoxin increases pulmonary vascular protein permeability in the dog

    International Nuclear Information System (INIS)

    Endotoxin increases pulmonary vascular permeability consistently in some species but fails to reliably cause injury in the dog. We wondered whether this phenomenon depended on the method of injury assessment, as others have relied on edema measurement; we quantified injury by monitoring the rate of extravascular protein accumulation. /sup 113m/In-labeled protein and /sup 99m/Tc-labeled erythrocytes were injected into anesthetized dogs and monitored by an externally placed lung probe. A protein leak index, the rate of extravascular protein accumulation, was derived from the rate of increase in lung protein counts corrected for changes in intravascular protein activity. After administration of Salmonella enteriditis endotoxin (4 micrograms/kg), the protein leak index was elevated 2.5-fold (41.1 +/- 4.6 X 10(-4) min-1) compared with control (16.0 +/- 2.8 X 10(-4) min-1). In contrast, wet-to-dry weight ratios failed to increase after endotoxin (4.6 +/- 0.8 vs. control values of 4.2 +/- 0.5 g/g dry bloodless lung). However, we observed that endotoxin increased lung dry weight (per unit body weight), which may have attenuated the change in wet-to-dry weight ratios. To determine whether low microvascular pressures following endotoxin attenuated edema formation, we increased pulmonary arterial wedge pressures in five dogs by saline infusion, which caused an increase in wet-to-dry weight ratios following endotoxin but no change in the five controls. We conclude that low dose endotoxin causes pulmonary vascular protein leak in the dog while edema formation is minimal or absent

  7. DETERMINATION OF ENDOTOXIN IN SMALL-VOLUME PARENTERALS (SVPs: A COMPARISON OF LAL GEL-CLOT METHOD WITH U.S.P. RABBIT PYROGEN TEST

    Directory of Open Access Journals (Sweden)

    Morteza Rafiee-Tehrani M.H. Djamshidi

    1994-08-01

    Full Text Available The simplicity and sensitivity of the Limulus Amebocyte Lysate (LAL gel-clot method in detection of endotoxin in Small Volume Parenterals (SVPs were studied and compared with rabbit pyrogen test."nAll formulations were artificially spiked with 0.25, 0.5 and 5EU/ml of Escherichia coli 0111:B4 endotoxin (Iambda=0.5EU/ml. The Threshold Pyrogenic Dose (TPD for E.coli 0111 :B4 endotoxin was found to be more than lng per kg of body weight (0.5EU/ml. However, the sensitivity of LAL test was found to be 0.25 Eu/ml for water for injection and 0.5 EU/ml for other formulations. The observed inhibitory effect of electrolyte containing solutions was deleted by some degree of dilution prior to test. However, none of the samples showed inhibitory at the Maximum Valid Dilution (MVD. The U.S.P. rabbit pyrogen test was insignificant for all the solutions tested. These results suggest, LAL gel-clot test to be useful for determination of bacterial endotoxin as final release test in SVPs. A comparison of the pyrogenicity assays also indicates, that LAL is more sensitive and simple and less time-consuming than the conventional rabbit test.

  8. Are Cats and Dogs the Major Source of Endotoxin in Homes?

    OpenAIRE

    Ownby, Dennis R.; Peterson, Edward L.; Wegienka, Ganesa; Woodcroft, Kimberley J.; Nicholas, Charlotte; Zoratti, Edward; Johnson, Christine C.

    2013-01-01

    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 wer...

  9. Effects of Postoperative Enteral Immune-enhancing Diet on Plasma Endotoxin Level, Plasma Endotoxin Inactivation Capacity and Clinical Outcome

    Institute of Scientific and Technical Information of China (English)

    YAO Guoxiang; XUE Xinbo; LU Xingpei; WANG Jianming; QIAN Jiaqin

    2005-01-01

    This study examined the postoperative plasma endotoxin level, plasma endotoxin inactivation capacity and clinical outcome after administration of an enteral diet supplemented with glutamine, arginine and ω-3-fatty acid in patients undergoing gastrointestinal operations on an prospective, randomized and double-blind design. 40 patients undergoing gastrointestinal operations were randomized into two groups, with each having 20 patients. One group received standard enteral nutrition and the other was fed the formulation supplemented with glutamine, arginine and ω-3-fatty acid. The two groups were isonitrogenous. The infusion was started from day 1 after surgery and continued for 7 days. Blood samples were collected on the morning of day 1 before operation and on the morning of 1, 4 and 7 day(s) after operation and analyzed for plasma endotoxin level and endotoxin inactivation capacity (EIC). Our study found no differences between the two groups on plasma endotoxin level. After surgery a rapid reduction in plasma endotoxin inactivation capacity was observed in both groups, a significant recovery of the plasma endotoxin inactivation capacity was observed on morning of day 4 after surgery in the study group (0.12±0.02 EU/mL and 0. 078±0.022 EU/mL respectively, P<0.01). Shortened hospital stay was observed in the experimental group (11.7±2.0 days in the control group and 10.6±1.2 days in the experimental group respectively, P=0.03). It is concluded that perioperative parenteral nutrition supplemented with glutamine, arginine and ω-3-fatty acid ameliorated postoperative immunodepression but without direct effect on endotoxemia.

  10. Harnessing aptamers for electrochemical detection of endotoxin.

    Science.gov (United States)

    Kim, Sung-Eun; Su, Wenqiong; Cho, MiSuk; Lee, Youngkwan; Choe, Woo-Seok

    2012-05-01

    Lipopolysaccharide (LPS), also known as endotoxin, triggers a fatal septic shock; therefore, fast and accurate detection of LPS from a complex milieu is of primary importance. Several LPS affinity binders have been reported so far but few of them have proved their efficacy in developing electrochemical sensors capable of selectively detecting LPS from crude biological liquors. In this study, we identified 10 different single-stranded DNA aptamers showing specific affinity to LPS with dissociation constants (K(d)) in the nanomolar range using a NECEEM-based non-SELEX method. Based on the sequence and secondary structure analysis of the LPS binding aptamers, an aptamer exhibiting the highest affinity to LPS (i.e., B2) was selected to construct an impedance biosensor on a gold surface. The developed electrochemical aptasensor showed excellent sensitivity and specificity in the linear detection range from 0.01 to 1 ng/mL of LPS with significantly reduced detection time compared with the traditional Limulus amoebocyte lysate (LAL) assay. PMID:22370280

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

    NARCIS (Netherlands)

    Chen, C.M.; Thiering, E.; Doekes, G.; Zock, J.P.; 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 E

  12. Respiratory effects of endotoxin exposure : Individual susceptibility and gene-environment interactions

    NARCIS (Netherlands)

    Smit, L.A.

    2008-01-01

    Endotoxins are cell wall components of Gram-negative bacteria that occur commonly on plants and plant products and as gut commensals. A large variability in airborne endotoxin exposure levels has been measured in a range of agricultural and other occupational environments. Inhalation of endotoxins m

  13. 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...

  14. 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...... Escherichia coli K12, probiotic E. coli Nissle, EPEC) induced basolateral production of TNF-alpha, IFN-gamma, IL 6, 8, and 10. Gram-positive probiotics (Lactobacillus spp. and Bifidobacterium spp.) had virtually no effect. In addition, commensals (Enterococcus faecalis, Bacteroides vulgatus) and food...... (polymyxin, colistin) completely abrogated transepithelial activation of leukocytes. Enterobacteriaceae-type endotoxin is a crucial factor in transepithelial stimulation of leukocytes, regardless of whether it is produced by probiotics or other bacteria. Hence, transepithelial stimulation ofleukocytes...

  15. 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...... Escherichia coli K12, probiotic E. coli Nissle, EPEC) induced basolateral production of TNF-alpha, IFN-gamma, IL 6, 8, and 10. Gram-positive probiotics (Lactobacillus spp. and Bifidobacterium spp.) had virtually no effect. In addition, commensals (Enterococcus faecalis, Bacteroides vulgatus) and food...... (polymyxin, colistin) completely abrogated transepithelial activation of leukocytes. Enterobacteriaceae-type endotoxin is a crucial factor in transepithelial stimulation of leukocytes, regardless of whether it is produced by probiotics or other bacteria. Hence, transepithelial stimulation of leukocytes...

  16. Resistance of genetically different common carp, Cyprinus carpio L., families against experimental bacterial challenge with Aeromonas hydrophila.

    Science.gov (United States)

    Jeney, G; Ardó, L; Rónyai, A; Bercsényi, M; Jeney, Z

    2011-01-01

    The objective of this study was to determine the differences in disease resistance against artificial infection with Aeromonas hydrophila between genetically different common carp families. Four strains differing in their origin and breeding history were selected from the live gene bank of common carp maintained at the Research Institute for Fisheries, Aquaculture and Irrigation (HAKI, Szarvas, Hungary) to establish families with wide genetic background: Szarvas 15 (15), an inbred mirror line; Tata (T) scaly noble carp; Duna (D), a Hungarian wild carp and Amur (A), an East Asian wild carp. A diallele mating structure was used to allow the assessment of genetic variation within and between the tested 96 families for a variety of traits. The existing technologies of fertilization and incubation of carp eggs, as well as larval and fingerling rearing had been modified because of the large number of baseline populations. Two challenge trials of the 96 families of carp with Aeromonas hydrophila were done. The 10 most resistant and 10 most susceptible families to A. hydrophila were identified from these two challenges. The crosses that produced the most resistant families were mainly those having parents from Tata and Szarvas 15 domesticated strains, while the most susceptible families were from the wild strains Duna and Amur.

  17. Immune response, stress resistance and bacterial challenge in juvenile rainbow trouts Oncorhynchus mykiss fed diets containing chitosan-oligosaccharides

    Directory of Open Access Journals (Sweden)

    Lin LUO, Xuefeng CAI, Chuan HE, Min XUE, Xiufeng WU , Haining CAO

    2009-12-01

    Full Text Available Effects of dietary supplementation of chitosan-oligosaccharides (COS on the growth performance, immune response, stress resistance, and disease resistance of juvenile rainbow trout Oncorhynchus mykiss were studied. Four experimental diets containing 0, 20, 40, or 60 mg/kg COS (COS0, COS20, COS40, and COS60, respectively were fed to juvenile rainbow trout (initial weight = 5.2 ± 0.3 g for 8 weeks. By the end of the feeding trial, representative groups of fish from each dietary treatment were challenged with stressor (30 sec air exposure and pathogen exposure (intraperitoneal injection with Aeromonas hydrophila. Results showed that supplementation of COS in diets did not affect production performance and body composition of rainbow trout. However, fish fed the COS40 diet demonstrated improved phagocytic activities, respiratory burst activities and decreased serum cortisol level. Additionally, survival following A. hydrophila challenge was significant higher among fish fed the COS-supplemented feeds, although there was no difference based on the level of supplementation. The present study suggests that COS can be used as an immuno-stimulant in rainbow trout feeds [Current Zoology 55 (6: 416– 422, 2009].

  18. Cloning and expression analysis of a ubiquitin gene (UbL40) in the haemocytes of Crassostrea hongkongensis under bacterial challenge

    Institute of Scientific and Technical Information of China (English)

    FU Dingkun; ZHANG Yang; YU Ziniu

    2011-01-01

    Ubiquitin, a highly conserved stress-related protein, is assigned multiple functions, such as DNA processing, protein degradation, and ribosome synthesis. The Crassostrea hongkongensis ubiquitin gene (designated ChUbL40) was cloned by a combination of suppressive subtractive hybridization (SSH) and rapid amplification of cDNA ends (RACE). The full-length cDNA of ChUbL40 is 496 bp in length, consisting of a 5' untranslated region (UTR) of 34 bp, a 3'-UTR of 75 bp and an open reading frame of 387 bp encoding a ubiquitin fusion protein of 128 amino acids. Analysis of the amino acid sequence of ChUbL40 reveals that UbL40 is highly conservative during evolution. The expression patterns of ChUbL40 gene in various tissues were examined by real-time PCR. The expression level of ChUbL40 in haemocytes is down-regulated at 4 h and gradually returned to its original level from 6 h to 24 h after Vibrio alginolyticus challenge. Our results suggest that ChUbL40 is ubiquitously expressed and plays an important role in immune defense against bacterial challenge.

  19. Size-fractionated PM10 monitoring in relation to the contribution of endotoxins in different polluted areas

    Science.gov (United States)

    Traversi, D.; Alessandria, L.; Schilirò, T.; Gilli, G.

    2011-07-01

    Particulate pollution is an environmental concern that is widespread and difficult to resolve. Recently various regulatory improvements around the world have been agreed upon to tackle this problem, especially as related to the fine fraction of particulates, which more closely correlates to human health effects than other fractions. The size-fractionation of inhalable particles and their organic composition represent a new area of research that has been poorly explored thus far. Endotoxins are a type of natural organic compound that can be found in particulate matter. They are correlated with Gram-negative bacterial contamination. Health outcomes associated with exposure to these toxins are not specific and often overlap with the health effects of PM (Particulate Matter) exposure, including asthma, bronchitis, acute respiratory distress syndrome and organic dust toxic syndrome. Very little information is available on the endotoxin distribution in different PM10 size fractions. This study examined PM10 size fractions and their endotoxin content. Sampling was conducted at five different locations: one urban, two rural and two rural sites that were highly influenced by large-scale farm animal production facilities. For each location, six different PM10 fractions were evaluated. PM10 sub-fractions were categorised as follows: PM 10-7.2 (1.15-31.30 μg m -3); PM 7.2-3.0 (1.86-30.73 μg m -3); PM 3.0-1.5 (1.74-13.90 μg m -3); PM 1.5-0.95 (0.24-10.57 μg m -3); PM 0.95-0.49 (1.22-14.33 μg m -3) and PM <0.49 (13.15-85.49 μg m -3). The ranges of endotoxin levels determined were: PM 10-7.2 (0.051-5.401 endotoxin units (EU) m -3); PM 7.2-3.0 (0.123-7.801 EU m -3); PM 3.0-1.5 (0.057-1.635 EU m -3); PM 1.5-0.95 (0.040-2.477 EU m -3); PM 0.95-0.49 (0.007-3.159 EU m -3) and PM <0.49 (0.039-3.975 EU m -3). Our results indicated consistency of the PM1 fraction at all of the sites and the predominant presence of endotoxins in the coarse fraction. The observed abatement of the PM

  20. 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

  1. Intramammary Immunization of Pregnant Mice with Staphylococcal Protein A Reduces the Post-Challenge Mammary Gland Bacterial Load but Not Pathology.

    Directory of Open Access Journals (Sweden)

    Jully Gogoi-Tiwari

    Full Text Available Protein A, encoded by the spa gene, is one of the major immune evading MSCRAMM of S. aureus, demonstrated to be prevalent in a significant percentage of clinical bovine mastitis isolates in Australia. Given its' reported significance in biofilm formation and the superior performance of S. aureus biofilm versus planktonic vaccine in the mouse mastitis model, it was of interest to determine the immunogenicity and protective potential of Protein A as a potential vaccine candidate against bovine mastitis using the mouse mastitis model. Pregnant Balb/c mice were immunised with Protein A emulsified in an alum-based adjuvant by subcutaneous (s/c or intramammary (i/mam routes. While humoral immune response of mice post-immunization were determined using indirect ELISA, cell-mediated immune response was assessed by estimation of interferon-gamma (IFN-γ produced by protein A-stimulated splenocyte supernatants. Protective potential of Protein A against experimental mastitis was determined by challenge of immunized versus sham-vaccinated mice by i/mam route, based upon manifestation of clinical symptoms, total bacterial load and histopathological damage to mammary glands. Significantly (p<0.05 higher levels of IgG1 isotype were produced in mice immunized by the s/c route. In contrast, significantly higher levels of the antibody isotype IgG2a were produced in mice immunized by the i/mam route (p<0.05. There was significant reduction (p<0.05 in bacterial loads of the mammary glands of mice immunized by Protein A regardless of the route of immunization, with medium level of clinical symptoms observed up to day 3 post-challenge. However, Protein A vaccine failed to protect immunized mice post-challenge with biofilm producing encapsulated S. aureus via i/mam route, regardless of the route of immunization, as measured by the level of mammary tissue damage. It was concluded that, Protein A in its' native state was apparently not a suitable candidate for inclusion

  2. α1-ANTITRYPSIN ATTENUATES ENDOTOXIN-INDUCED ACUTE LUNG INJURY IN RABBITS

    Institute of Scientific and Technical Information of China (English)

    揭志军; 蔡映云; 杨文兰; 金美玲; 朱威; 祝慈芳

    2003-01-01

    Objective To investigate whether pretreatment with α1-AT can attenuate acute lung injury (ALI) in rabbits induced with endotoxin. Methods Thirty-two New Zealand rabbits were randomly assigned to four groups(n=8):1.Infusion of endotoxin(Lipopolysaccharide,LPS 500μg/kg)without α1-AT (group LPS).2.Infusion α1-AT 120mg/kg at 15min before challenge with LPS(group LAV).3.Infusion of α1-AT 120mg/kg(group AAT).4 Infusion of saline 4ml/kg as control (group NS).Arterial blood gases,peripheral leukocyte counts and airway pressure were recorded every 1h.Physiologic intrapulmonary shunting (Qs/Qt) was measured every 4h.After 8h the bloods were collected for measurement of plasma concentration and activity of α1-AT.Then bronchoalveolar lavage fluid (BALF)was collected for measurement of concentrations of total protein (TP),interleukin-8(IL-8),tumor necrosis factor(TNF-α),the activities of elastase-like and α1-AT,total phospholipids(TPL) and disaturated phosphatidylcholine (DSPC).In addition,the wet-to-dry lung weight ratio(W/D) was measured. Results After infusion of endotoxin,it was observed that PaO2,peripheral luekocyte counts,total respiratory compliance progressively decreased and Ppeak and Qs/Qt increased comparing with the baseline values.In contrast to group NS,the increased plasma concentration but reduced activity of α1-AT was found in group LPS.In the BALF,the activity of α1-AT,TPL,DSPC/TPL were lower,but the concentrations of albumin,IL-8,TNF-α,and the activity of NE were higher.The ratio of W/D also increased.The pretreatment of α1-AT attenuated the deterioration of oxygenation,the reduction of compliance and the deterioration of other physiological,biochemical parameters mentioned above. Conclusion Pretreatment with α1-AT could attenuate endotoxin-induced lung injury in rabbits.Those beneficial effects of α1-AT might be due in part to the inhibitory effect on neutrophil elastase.

  3. Metal ions potentiate microglia responsiveness to endotoxin.

    Science.gov (United States)

    Rachmawati, Dessy; Peferoen, Laura A N; Vogel, Daphne Y S; Alsalem, Inás W A; Amor, Sandra; Bontkes, Hetty J; von Blomberg, B Mary E; Scheper, Rik J; van Hoogstraten, Ingrid M W

    2016-02-15

    Oral metal exposure has been associated with diverse adverse reactions, including neurotoxicity. We showed previously that dentally applied metals activate dendritic cells (MoDC) via TLR4 (Ni, Co, Pd) and TLR3 (Au). It is still unknown whether the low levels of dental metals reaching the brain can trigger local innate cells or prime them to become more responsive. Here we tested whether dentally applied metals (Cr, Fe, Co, Ni, Cu, Zn, Au, Hg) activate primary human microglia in vitro and, as a model, monocytic THP-1-cells, in high non-toxic as well as near-physiological concentrations. In addition the effects of 'near-physiological' metal exposure on endotoxin (LPS) responsiveness of these cells were evaluated. IL-8 and IL-6 production after 24h was used as read out. In high, non-toxic concentrations all transition metals except Cr induced IL-8 and IL-6 production in microglia, with Ni and Co providing the strongest stimulation. When using near-physiological doses (up to 10× the normal plasma concentration), only Zn and Cu induced significant IL-8 production. Of note, the latter metals also markedly potentiated LPS responsiveness of microglia and THP-1 cells. In conclusion, transition metals activate microglia similar to MoDCs. In near-physiological concentrations Zn and Cu are the most effective mediators of innate immune activation. A clear synergism between innate responses to Zn/Cu and LPS was observed, shedding new light on the possible relation between oral metal exposure and neurotoxicity.

  4. Programmed cell death 4 in bacterially-challenged Apostichopus japonicus: Molecular cloning, expression analysis and functional characterization.

    Science.gov (United States)

    Lv, Zhimeng; Li, Chenghua; Shao, Yina; Zhang, Weiwei; Wang, Zhenhui; Wang, Haihong

    2016-07-01

    Programmed cell death 4 (PDCD4) plays a crucial role in modulating cellular signals, mainly via TOLL cascades during the immune response. In the present study, a novel PDCD4 homologue gene (denoted as AjPDCD4) was cloned from the sea cucumber Apostichopus japonicus using RACE. The full-length AjPDCD4 cDNA comprised a 366bp 5'-UTR, a 418bp 3'-UTR, and a 1353bp open reading frame encoding a 450 amino acid residue protein with two typical MA3 domains. Phylogenetic analysis revealed that AjPDCD4 belonged to the invertebrate PDCD4 family. Spatial expression analysis indicated that AjPDCD4 mRNA transcripts are expressed at a high level in the tentacles and at a low level in muscle compared with coelomocytes. Vibrio splendidus challenge and LPS exposure could both significantly down-regulate AjPDCD4 mRNA expression. More importantly, we found that ultraviolet (UV)-induced ROS production and DNA damage were greatly repressed in AjPDCD4-knockdown coelomocytes. Meanwhile, the expression levels of the NF-kappa B homologue, p105, were synchronously up-regulated in the same conditions. All of these results indicated that AjPDCD4 is involved in modulating DNA damage and ROS production in sea cucumber, perhaps by affecting the TLR pathway. PMID:27262523

  5. Cattle Immunized with a Recombinant Subunit Vaccine Formulation Exhibits a Trend towards Protection against Histophilus somni Bacterial Challenge

    Science.gov (United States)

    Madampage, Claudia Avis; Wilson, Don; Townsend, Hugh; Crockford, Gordon; Rawlyk, Neil; Dent, Donna; Evans, Brock; Van Donkersgoed, Joyce; Dorin, Craig; Potter, Andrew

    2016-01-01

    Histophilosis, a mucosal and septicemic infection of cattle is caused by the Gram negative pathogen Histophilus somni (H. somni). As existing vaccines against H. somni infection have shown to be of limited efficacy, we used a reverse vaccinology approach to identify new vaccine candidates. Three groups (B, C, D) of cattle were immunized with subunit vaccines and a control group (group A) was vaccinated with adjuvant alone. All four groups were challenged with H. somni. The results demonstrate that there was no significant difference in clinical signs, joint lesions, weight change or rectal temperature between any of the vaccinated groups (B,C,D) vs the control group A. However, the trend to protection was greatest for group C vaccinates. The group C vaccine was a pool of six recombinant proteins. Serum antibody responses determined using ELISA showed significantly higher titers for group C, with P values ranging from < 0.0148 to < 0.0002, than group A. Even though serum antibody titers in group B (5 out of 6 antigens) and group D were significantly higher compared to group A, they exerted less of a trend towards protection. In conclusion, the vaccine used in group C exhibits a trend towards protective immunity in cattle and would be a good candidate for further analysis to determine which proteins were responsible for the trend towards protection. PMID:27501390

  6. Bacteria endotoxin test of Vitamin B Co. Injection%复合维生素B注射液细菌内毒素检查法的研究

    Institute of Scientific and Technical Information of China (English)

    刘骅; 武谷

    2011-01-01

    Aim To establish the bacterial endotoxin test for Vitamin B Co. Injection. Methods The experiment was carried out according to the appendix of Chinese Pharmacopeia in 2005 Edition (Vol Ⅱ ). Results The reactions with two Tachypleus Amebocyte Lysate manufacturers were not disturbed in minimus diluted multiple for Vitamin B Co. Injection,which was one ratio sixteen. The endotoxin limit of Vitamin B Co. Injection was 5 EU · ml -1. Conclusion It is feasible to control quality of Vitamin B Co. Injection by bacterial endotoxin test.%目的 建立复合维生素B注射液的细菌内毒素检查方法.方法 根据2005年版二部附录细菌内毒素检查法,采用2个不同生产厂家的鲎试剂对复合维生素B注射液进行研究.结果 复合维生素B注射液最小不干扰稀释倍数为1: 16,细菌内毒素限值为5 EU·ml-1.结论 采用细菌内毒素检查法控制复合维生素B注射液的质量是可行的.

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

    Directory of Open Access Journals (Sweden)

    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.

  8. Neurons and astroglia govern microglial endotoxin tolerance through macrophage colony-stimulating factor receptor-mediated ERK1/2 signals.

    Science.gov (United States)

    Chu, Chun-Hsien; Wang, Shijun; Li, Chia-Ling; Chen, Shih-Heng; Hu, Chih-Fen; Chung, Yi-Lun; Chen, Shiou-Lan; Wang, Qingshan; Lu, Ru-Band; Gao, Hui-Ming; Hong, Jau-Shyong

    2016-07-01

    Endotoxin tolerance (ET) is a reduced responsiveness of innate immune cells like macrophages/monocytes to an endotoxin challenge following a previous encounter with the endotoxin. Although ET in peripheral systems has been well studied, little is known about ET in the brain. The present study showed that brain immune cells, microglia, being different from peripheral macrophages, displayed non-cell autonomous mechanisms in ET formation. Specifically, neurons and astroglia were indispensable for microglial ET. Macrophage colony-stimulating factor (M-CSF) secreted from these non-immune cells was essential for governing microglial ET. Neutralization of M-CSF deprived the neuron-glia conditioned medium of its ability to enable microglia to form ET when microglia encountered two lipopolysaccharide (LPS) treatments. Recombinant M-CSF protein rendered enriched microglia refractory to the second LPS challenge leading to microglial ET. Activation of microglial M-CSF receptor (M-CSFR; also known as CSF1R) and the downstream ERK1/2 signals was responsible for M-CSF-mediated microglial ET. Endotoxin-tolerant microglia in neuron-glia cultures displayed M2-like polarized phenotypes, as shown by upregulation of M2 marker Arg-1, elevated production of anti-inflammatory cytokine interleukin 10, and decreased secretion of pro-inflammatory mediators (tumor necrosis factor α, nitric oxide, prostaglandin E2 and interleukin 1β). Endotoxin-tolerant microglia protected neurons against LPS-elicited inflammatory insults, as shown by reduced neuronal damages in LPS pre-treatment group compared with the group without LPS pre-treatment. Moreover, while neurons and astroglia became injured during chronic neuroinflammation, microglia failed to form ET. Thus, this study identified a distinct non-cell autonomous mechanism of microglial ET. Interactions of M-CSF secreted by neurons and astroglia with microglial M-CSFR programed microglial ET. Loss of microglial ET could be an important

  9. Role of intestinal bifidobacteria in the pathogenesis of gut-derived bacteria/endotoxin translocation and the effect of bifidobacterial supplement on gut barrier function following burns

    Institute of Scientific and Technical Information of China (English)

    Wang Zhongtang; Yao Yongming; Xiao Guangxia; Sheng Zhiyong

    2003-01-01

    Early multiple organ dysfunction syndrome appears to be facilitated with bacterial translocation in severely burn injury, yet the mechanisms of bacterial translocation remains in dispute. The aim of this study was to investigate the potential role of intestinal bifidobacteria in the pathogenesis of gut-derived bacteria/endotoxin translocation following burns and the effects of bifidobacterial supplement on gut barrier. Methods: Wistar rats were randomly divided into burn group (Burn, n=60),sham burn group (SB, n=10) in experiment Ⅰ , and burn + saline group (BS, n=30), burn + bifidobacteria group (BB, n=30), and sham-burn + saline group (SS, n= 10) in experiment Ⅱ. Animals in BB group were fed bifidobacterial preparation (5 × 109 CFU/ml) after burns, 1.5ml,twice daily. Animals in BS and SS were fed saline. Samples were taken on days 1, 3, and 5 in burn groups, and on day 3 in sham-burn groups. The incidence of bacteria/endotoxin translocation and counts of Bifidobacterium, Fungi and Escherichia coli in gut mucosa were determined with standard methods. The levels of sIgA in mucus of small intestine were measured by RIA. The positive sIgA expression in lamina propria and ileum mucosal injury was evaluated light microscopically by blinded examiners. Results: Our results showed that the incidence of bacterial translocation was increased after burns, which was accompanied by significant decrease in number of bifidobacteria but significant increase in E. coli and fungi in gut mucosa, and elevation of levels of plasma endotoxin and IL-6 (P<0. 001).The incidence of bacterial translocation was markedly reduced after 3- and 5-day supplementation of bifidobacteria compared with control group (P<0.05). The counts of mucosal bifidobacteria were increased by 4- to 40-fold,while E. coli and fungi were decreased by 2- to 30-fold and 10- to 150-fold, respectively, after bifidobacterial supplementation in contrast to control group. The damage of mucosa tended to be less

  10. Bacteria Endotoxin Test of Vitamin B6 for Injection%注射用维生素B6细菌内毒素检查法的研究

    Institute of Scientific and Technical Information of China (English)

    刘骅

    2012-01-01

    OBJECTIVE To establish the bacterial endotoxin test of vitamin B6 for injection. METHODS The experiment was carried out according to the appendix of ChP 2010 Vol Ⅱ. tachypleus amabocyte lysate from two manufacturers were used to test the bacterial endotoxin. RESULTS The endotoxin limit of vitamin B6 for injection was 0.30 EU·mg‐1. The reactions were not disturbed in 0.208 mg·mL‐1 for vitamin B6, which was the maximum nondisrurbing concentration. It was effective that endotoxin was tested by amebocyte lysate agent with the sensitivity of 0.06 EU·mL ‐1 or much higher. CONCLUSION ft is feasible to control quality of vitamin B6 for injection by bacterial endotoxin test.%目的 建立注射用维生素B6的细菌内毒素检查方法.方法 根据中国药典2010年版二部附录细菌内毒素检查法,采用2个不同厂家的鲎试剂对注射用维生素B6进行研究.结果 注射用维生素B6细菌内毒素检查限值为0.30 EU·mg-1,其最大不干扰浓度为0.208 mg·mL-1,用灵敏度为0.06 EU·mL-1或更高灵敏度的鲎试剂可进行检验.结论 细菌内毒素检查法可用于注射用维生素B6中内毒素的检查.

  11. 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. PMID:27470947

  12. Endotoxin levels in homes and classrooms of Dutch school children and respiratory health.

    Science.gov (United States)

    Jacobs, José H; Krop, Esmeralda J M; de Wind, Siegfried; Spithoven, Jack; Heederik, Dick J J

    2013-08-01

    Several studies describe indoor pollutant exposure in homes and to a lesser extent in schools. Population studies that include both environments are sparse. This study aims to assess endotoxin levels in primary schools and homes of children. Endotoxin was also studied in relation to asthma and sensitisation. 10 schools with (index) and without (reference) dampness were selected, based on reports and inspections. Cases and controls were selected from 169 homes based on the presence or absence of asthma-like symptoms of children. Classroom and bedroom airborne settled dust was sampled using electrostatic dust fall collectors. Average endotoxin levels in schools ranged from 2178 to 6914 endotoxin units (EU)·m(-2) per week compared with 462-1285 EU·m(-2) per week in homes. After mutual adjustment for home and school endotoxin, school endotoxin was positively associated with nonatopic asthma (OR 1.11, 95% CI 0.97-1.27), while no associations with endotoxin were found at home. The high endotoxin levels in schools compared with homes indicate that exposure at school can contribute considerably to environmental endotoxin exposure of children and teachers. Our results also suggest that endotoxin in schools may be associated with nonatopic asthmatic symptoms in pupils, although the results require reproduction because of the modest sample size. PMID:23100494

  13. Biophysical characterization of the interaction of Limulus polyphemus endotoxin neutralizing protein with lipopolysaccharide.

    Science.gov (United States)

    Andrä, Jörg; Garidel, Patrick; Majerle, Andreja; Jerala, Roman; Ridge, Richard; Paus, Erik; Novitsky, Tom; Koch, Michel H J; Brandenburg, Klaus

    2004-05-01

    Endotoxin-neutralizing protein (ENP) of the horseshoe crab is one of the most potent neutralizers of endotoxins [bacterial lipopolysaccharide (LPS)]. Here, we report on the interaction of LPS with recombinant ENP using a variety of physical and biological techniques. In biological assays (Limulus amebocyte lysate and tumour necrosis factor-alpha induction in human mononuclear cells), ENP causes a strong reduction of the immunostimulatory ability of LPS in a dose-dependent manner. Concomitantly, the accessible negative surface charges of LPS and lipid A (zeta potential) are neutralized and even converted into positive values. The gel to liquid crystalline phase transitions of LPS and lipid A shift to higher temperatures indicative of a rigidification of the acyl chains, however, the only slight enhancement of the transition enthalpy indicates that the hydrophobic moiety is not strongly disturbed. The aggregate structure of lipid A is converted from a cubic into a multilamellar phase upon ENP binding, whereas the secondary structure of ENP does not change due to the interaction with LPS. ENP contains a hydrophobic binding site to which the dye 1-anilino-8-sulfonic acid binds at a K(d) of 19 micro m, which is displaced by LPS. Because lipopolysaccharide-binding protein (LBP) is not able to bind to LPS when ENP and LPS are preincubated, tight binding of ENP to LPS can be deduced with a K(d) in the low nonomolar range. Importantly, ENP is able to incorporate by itself into target phospholipid liposomes, and is also able to mediate the intercalation of LPS into the liposomes thus acting as a transport protein in a manner similar to LBP. Thus, LPS-ENP complexes might enter target membranes of immunocompetent cells, but are not able to activate due to the ability of ENP to change LPS aggregates from an active into an inactive form. PMID:15128313

  14. Determination of water quality variables, endotoxin concentration, and Enterobacteriaceae concentration and identification in southern High Plains dairy lagoons.

    Science.gov (United States)

    Purdy, C W; Clark, R N; Straus, D C

    2010-04-01

    The objectives of this study were to determine the concentration of endotoxin, determine 20 water quality variables, and identify and enumerate fungal and bacterial pathogens from United States southern High Plains dairy lagoons and control lakes during summer and winter. Water samples were collected in triplicate from the north, south, east, and west quadrants of each body of water. The mean (+/- SEM) winter dairy lagoon endotoxin concentration was significantly higher (9,678+/-1,834 ng/mL) than the summer concentration (3,220+/-810 ng/mL). The mean endotoxin concentration of the 2 control lakes (summer: 58.1+/-8.8 ng/mL; winter: 38.6+/-4.2 ng/mL) was significantly less than that of the dairy lagoons. Two hundred-one Salmonella enterica spp. isolates were identified, 7 serovars were recovered from the dairy lagoons, and 259 Salmonella ssp. were identified from 5 other dairy locations (milk barn, ditch effluent, settling basin, feed alley pad flush, and center pivots). Twenty-eight Salmonella spp. were identified from center pivot water. Escherichia coli O157:H7 pathogens were isolated from other dairy locations but not from lagoons. Neither Salmonella spp. nor E. coli O157:H7 were identified from control lakes. Enterobacteriaceae opportunistic pathogens were isolated from both dairies and control lakes. Important mesophilic and thermophilic catabolic (to manure biosolids) fungal isolates were identified from dairy effluent locations, but no thermophilic fungal isolates were cultured from the control lakes. Adequate curing of green forage following center pivot irrigation is important to kill lagoon water enteric pathogens, even though the lagoon water is mixed with fresh water. Recirculating lagoon water to flush the feed alley pad, where cows stand while eating, to remove manure and using lagoon water to abate dairy dust in loafing pens and unimproved dairy roads is inconsistent with good environmental practice management. PMID:20338428

  15. Bacterial translocation in D-galactosamine-treated rats in a burn model

    NARCIS (Netherlands)

    Dijkstra, HM; Manson, WL; Blaauw, B; Klasen, HJ; deSmet, B

    1996-01-01

    Bacteria and endotoxins can pass through the gut barrier under certain conditions. This process of bacterial translocation (BT) may occur after thermal injury in animals and is thought to play a role in the pathogenesis of septic complications in severely burned patients. The current study was perfo

  16. Personal Exposure to Dust and Endotoxin in Robusta and Arabica Coffee Processing Factories in Tanzania

    OpenAIRE

    Sakwari, Gloria; Mamuya, Simon H D; Bråtveit, Magne; Larsson, Lennart; Pehrson, Christina; Moen, Bente E

    2013-01-01

    Introduction: 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.Methods: Using Sidekick Casella pumps at a flow rate of 2l/min, total ...

  17. 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...

  18. Masking of endotoxin in surfactant samples: Effects on Limulus-based detection systems.

    Science.gov (United States)

    Reich, Johannes; Lang, Pierre; Grallert, Holger; Motschmann, Hubert

    2016-09-01

    Over the last few decades Limulus Amebocyte Lysate (LAL) has been the most sensitive method for the detection of endotoxins (Lipopolysaccharides) and is well accepted in a broad field of applications. Recently, Low Endotoxin Recovery (LER) in biopharmaceutical drug products has been noticed, whereby the detection of potential endotoxin contaminations is not ensured. Notably, most of these drug products contain surfactants, which can have crucial effects on the detectability of endotoxin. In order to analyze the driving forces of LER, endotoxin detection in samples containing nonionic surfactants in various buffer systems was investigated. The results show that the process of LER is kinetically controlled and temperature-dependent. Furthermore, only the simultaneous presence of nonionic surfactants and components capable of forming metal complexes resulted in LER. In addition, capacity experiments show that even hazardous amounts of endotoxin can remain undetectable within such formulation compositions. In conclusion, the LER phenomenon is caused by endotoxin masking and not by test interference. In this process, the supramolecular structure of endotoxin is altered and exhibits only a limited susceptibility in binding to the Factor C of Limulus-based detection systems. We propose a two-step mechanism of endotoxin masking by complex forming agents and nonionic surfactants. PMID:27464990

  19. Effects of Endotoxin on Liver Smac Apoptosis Channel

    Institute of Scientific and Technical Information of China (English)

    Miao CHEN; Jian ZI-IOU; Hui LI; Anqun CHEN; Zhengang ZHANG; Deying TIAN

    2008-01-01

    To study the effect of endotoxin on liver apoptosis, LO2 liver ceils were cultured and passaged in vitro, and then stimulated by endotoxin at 10 mg/mL for 4, 8, 16 and 24 h respectively. Liver apoptosis was flow cytometrically and fluorescently detected. Immunohistochemistry was used to detect the delivery of smac and caspase9. The delivery of liver cell smac and the activity of caspase3 were measured by caspase3 assay kit. The hepatic failure models of rats were established by using D-galactosamine. The blood serum and liver tissues were collected for the detection of the liver function, the level of endotoxin and the activity of caspase3 by using chromogenic substrate limulus amebocyte lysate method (LAL) and caspase3 active assay kit. The expression of smac and caspase9 in liver ceils was detected by Western blotting. With in vitro study, the LO2 cells stimulated by LPS condensed into conglobation and formed apoptotic bodies. After those cells were stained.by hoechst,the apoptotic cells displayed blue color under the fluorescent microscope. The apoptosis rate was increased over time and the apoptosis was mainly of advanced stage. Meanwhile, the rate of smac delivery and activity of caspase9 and caspase3 were increased on LO2 cell membrane. In vivo, hepatic failure and obvious endotoxemia were induced by injection of more than 200 mg/kg D-GaIN. Hepatic mitochondria smac was reduced with dosage of D-GaIN and, on the contrary, the activity of caspase3 was increased. D-GaIN at 200 mg/kg increased Caspase9 while D-GaIN at 300 mg/kg decreased caspase9. Mitochondria signal channel plays an important role in the endotoxin-induced apoptosis of hepatic cells by promoting the release of smac from mitochondria to cytoplasm and activating caspase9 and caspase3 in its low-level channel.

  20. 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. PMID:14563656

  1. 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 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. PMID:25225707

  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. 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

  4. Liver Cirrhosis and Intestinal Bacterial Translocation

    Institute of Scientific and Technical Information of China (English)

    2014-01-01

    Intestinal barrier dysfunction, facilitating translocation of bacteria and bacterial products, plays an important role in the pathophysiology of liver cirrhosis and its complications. Intestinal defense system including microbial barrier, immunologic barrier, mechanical barrier, chemical barrier, plays an important role in the maintenance of intestinal function. Under normal circumstances, the intestinal barrier can prevent intestinal bacteria through the intestinal wall from spreading to the body. Severe infection, trauma, shock, cirrhosis, malnutrition, immune suppression conditions, intestinal bacteria and endotoxin translocation, can lead to multiple organ dysfunction. The intestinal microlfora is not only involved in the digestion of nutrients, but also in local immunity, forming a barrier against pathogenic microorganisms. The derangement of the gut microlfora may lead to microbial translocation, deifned as the passage of viable microorganisms or bacterial products from the intestinal lumen to the mesenteric lymph nodes and other extraintestinal sites. In patients with cirrhosis, primary and intestinal lfora imbalance, intestinal bacterial overgrowth, intestinal mucosal barrier dysfunction, endotoxemia is associated with weakened immunity.

  5. 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.

  6. Airborne Endotoxin from Indoor and Outdoor Environments:Effect of Sample Dilution on the Kinetic Limulus Amebocyte Lysate (LAL) Assay

    Science.gov (United States)

    Airborne endotoxin in occupational environments are a potential respiratory hazard to individuals. In this study, total and inhalable airborne endotoxin samples were collected via filtration from inside animal housing units and downwind from agricultural production sites and a wastewater treatment ...

  7. Effects of in vivo 'priming' on endotoxin-induced hypotension and tissue injury. The role of PAF and tumor necrosis factor.

    OpenAIRE

    Sun, X. M.; Hsueh, W; Torre-Amione, G.

    1990-01-01

    Exogenously administered tumor necrosis factor-alpha (TNF) and bacterial endotoxin (LPS) induce shock and tissue injury. Here, the authors studied the effect of endogenous TNF on LPS-induced hypotension and tissue injury and investigated the role of PAF in these responses. Rats were primed with intraperitoneal injection of zymosan 24 hours before, or Bacillus Calmette-Guérin (BCG) 12 to 15 days before intravenous injection of low dose (0.5 mg/kg) LPS. It was found that nonprimed animals showe...

  8. Effect of Tris-acetate buffer on endotoxin removal from human-like collagen used biomaterials

    International Nuclear Information System (INIS)

    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

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

    Energy Technology Data Exchange (ETDEWEB)

    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.

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

    International Nuclear Information System (INIS)

    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

  11. Bacterial Hydrodynamics

    Science.gov (United States)

    Lauga, Eric

    2016-01-01

    Bacteria predate plants and animals by billions of years. Today, they are the world's smallest cells, yet they represent the bulk of the world's biomass and the main reservoir of nutrients for higher organisms. Most bacteria can move on their own, and the majority of motile bacteria are able to swim in viscous fluids using slender helical appendages called flagella. Low-Reynolds number hydrodynamics is at the heart of the ability of flagella to generate propulsion at the micrometer scale. In fact, fluid dynamic forces impact many aspects of bacteriology, ranging from the ability of cells to reorient and search their surroundings to their interactions within mechanically and chemically complex environments. Using hydrodynamics as an organizing framework, I review the biomechanics of bacterial motility and look ahead to future challenges.

  12. Bacterial hydrodynamics

    CERN Document Server

    Lauga, Eric

    2015-01-01

    Bacteria predate plants and animals by billions of years. Today, they are the world's smallest cells yet they represent the bulk of the world's biomass, and the main reservoir of nutrients for higher organisms. Most bacteria can move on their own, and the majority of motile bacteria are able to swim in viscous fluids using slender helical appendages called flagella. Low-Reynolds-number hydrodynamics is at the heart of the ability of flagella to generate propulsion at the micron scale. In fact, fluid dynamic forces impact many aspects of bacteriology, ranging from the ability of cells to reorient and search their surroundings to their interactions within mechanically and chemically-complex environments. Using hydrodynamics as an organizing framework, we review the biomechanics of bacterial motility and look ahead to future challenges.

  13. 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 effects...tive endotoxin: an extraordinary lipid with profound effects oneukaryotic signal transduction. Authors Raetz

  14. 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.

  15. 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....

  16. Relationship between chicken cellular immunity and endotoxin levels in dust from chicken housing environments.

    Science.gov (United States)

    Roque, Katharine; Shin, Kyung-Min; Jo, Ji-Hoon; Kim, Hyoung-Ah; Heo, Yong

    2015-01-01

    Hazardous biochemical agents in animal husbandry indoor environments are known to promote the occurrence of various illnesses among workers and animals. The relationship between endotoxin levels in dust collected from chicken farms and various immunological markers was investigated. Peripheral blood was obtained from 20 broiler chickens and 20 laying hens from four different chicken farms in Korea. Concentrations of total or respirable dust in the inside the chicken farm buildings were measured using a polyvinyl chloride membrane filter and mini volume sampler. Endotoxin levels in the dust were determined by the Limulus Amebocyte Lysate Kinetic method. Interferon-γ production by peripheral blood mononuclear cells stimulated with concanavalin A was significantly lower in broilers or layers from the farms with higher endotoxin concentrations than the chickens from the farms with lower endotoxin levels. An opposite pattern was observed for plasma cortisol concentrations with higher cortisol levels found in chickens from the farms with higher endotoxin levels. When peripheral lymphocytes were examined, the percentage of CD3(-)Ia(+) B cells was lower in layers from farms with higher endotoxin levels than those from locations with lower endotoxin levels. Overall, these results suggest a probable negative association between dust endotoxin levels and cell-mediated immunity in chickens.

  17. Some immuno-suppressive trends: Effects Of Endotoxin on Camels (Camelus dromedarius)

    International Nuclear Information System (INIS)

    Intravenous administration of a bolus of 0.05 micron g/kg body weight endotoxin lipopolysaccharide (LPS) to camels has resulted in leukopenia, lymphopenia, neutropenia, proteinaemia, albunaemia , globulinaemia and reduced lysozomal enzyme activity. It is suggested that endotoxin at the dose of 0.05micron g/kg body weight may produce immunosuppressive effects in camels. (author)

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

    Energy Technology Data Exchange (ETDEWEB)

    Layman, D.L.; Diedrich, D.L.

    1987-06-01

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

  19. 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 Twe

  20. Electrochemical endotoxin sensors based on TLR4/MD-2 complexes immobilized on gold electrodes.

    Science.gov (United States)

    Yeo, Tae Yun; Choi, Ji Suk; Lee, Byung Kook; Kim, Beob Soo; Yoon, Hwa In; Lee, Hyeong Yun; Cho, Yong Woo

    2011-10-15

    Even low concentrations of endotoxins can be life-threatening. As such, continuous effort has been directed toward the development of sensitive and specific endotoxin detection systems. In this paper, we report the design and fabrication of a new electrochemical endotoxin sensor based on a human recombinant toll-like receptor 4 (rhTLR4) and myeloid differentiation-2 (MD-2) complex. The rhTLR4/MD-2 complex, which specifically binds to endotoxin, was immobilized on gold electrodes through a self-assembled monolayer (SAM) technique involving the use of dithiobis(succinimidyl undecanoate) (DSU). The surface topography of the electrodes at each fabrication stage was characterized with a nanosurface profiler and atomic force microscope (AFM). The electrochemical signals generated from interactions between the rhTLR4/MD-2 complex and the endotoxin were characterized by cyclic voltammetry (CV) and differential pulse voltammetry (DPV). A linear relationship between the peak current and endotoxin concentration was obtained in the range of 0.0005 to 5 EU/mL with a correlation coefficient (R(2)) of 0.978. The estimated limit of detection (LOD) was fairly low, 0.0002 EU/mL. The rhTLR4/MD-2 based sensors exhibited no current responses to dipalmitoylphosphatidylcholine (DPPC) bearing two lipid chains, which is structurally similar to endotoxin, indicating the high specificity of the sensors to endotoxin. PMID:21816600

  1. [Development of a piezoelectric sensor for detection of low endotoxin concentration].

    Science.gov (United States)

    Xiong, Xingliang; Wang, Xiang; Cai, Shaoxi

    2005-10-01

    Endotoxin is an important factor which can lead to endotoxemia and complication. Accurate detection of its concentration is very useful for the diagnosis and treatment of these diseases. A piezoelectric biosensor for detecting endotoxin was developed, which was based on liquid damping effect of quartz crystal resonator. The test results showed that the maximal frequency shift of sensor is linearly dependent on the logarithm value of concentration of endotoxin (0.1 pg/m - 10 ng/ml). The time which d (deltaf)/dt(maax) appeared in frequency shift curve was also linearly dependent on the logarithm value of concentration of endotoxin (0.01 pg/ml - 10 ng/ml). The detection time was shortened and the minimal limit of detection was decreased using the second method. Thus the proposed sensor is much simpler, more precise and has more lower limit of detecting detection of endotoxin when compared with the conventional methods. PMID:16294749

  2. Anthrapyrazolone analogues intercept inflammatory JNK signals to moderate endotoxin induced septic shock

    Science.gov (United States)

    Prasad, Karothu Durga; Trinath, Jamma; Biswas, Ansuman; Sekar, Kanagaraj; Balaji, Kithiganahalli N.; Guru Row, Tayur N.

    2014-11-01

    Severe sepsis or septic shock is one of the rising causes for mortality worldwide representing nearly 10% of intensive care unit admissions. Susceptibility to sepsis is identified to be mediated by innate pattern recognition receptors and responsive signaling pathways of the host. The c-Jun N-terminal Kinase (JNK)-mediated signaling events play critical role in bacterial infection triggered multi-organ failure, cardiac dysfunction and mortality. In the context of kinase specificities, an extensive library of anthrapyrazolone analogues has been investigated for the selective inhibition of c-JNK and thereby to gain control over the inflammation associated risks. In our comprehensive biochemical characterization, it is observed that alkyl and halogen substitution on the periphery of anthrapyrazolone increases the binding potency of the inhibitors specifically towards JNK. Further, it is demonstrated that hydrophobic and hydrophilic interactions generated by these small molecules effectively block endotoxin-induced inflammatory genes expression in in vitro and septic shock in vivo, in a mouse model, with remarkable efficacies. Altogether, the obtained results rationalize the significance of the diversity oriented synthesis of small molecules for selective inhibition of JNK and their potential in the treatment of severe sepsis.

  3. Short-term dynamics of indoor and outdoor endotoxin exposure: Case of Santiago, Chile, 2012.

    Science.gov (United States)

    Barraza, Francisco; Jorquera, Héctor; Heyer, Johanna; Palma, Wilfredo; Edwards, Ana María; Muñoz, Marcelo; Valdivia, Gonzalo; Montoya, Lupita D

    2016-01-01

    Indoor and outdoor endotoxin in PM2.5 was measured for the very first time in Santiago, Chile, in spring 2012. Average endotoxin concentrations were 0.099 and 0.094 [EU/m(3)] for indoor (N=44) and outdoor (N=41) samples, respectively; the indoor-outdoor correlation (log-transformed concentrations) was low: R=-0.06, 95% CI: (-0.35 to 0.24), likely owing to outdoor spatial variability. A linear regression model explained 68% of variability in outdoor endotoxins, using as predictors elemental carbon (a proxy of traffic emissions), chlorine (a tracer of marine air masses reaching the city) and relative humidity (a modulator of surface emissions of dust, vegetation and garbage debris). In this study, for the first time a potential source contribution function (PSCF) was applied to outdoor endotoxin measurements. Wind trajectory analysis identified upwind agricultural sources as contributors to the short-term, outdoor endotoxin variability. Our results confirm an association between combustion particles from traffic and outdoor endotoxin concentrations. For indoor endotoxins, a predictive model was developed but it only explained 44% of endotoxin variability; the significant predictors were tracers of indoor PM2.5 dust (Si, Ca), number of external windows and number of hours with internal doors open. Results suggest that short-term indoor endotoxin variability may be driven by household dust/garbage production and handling. This would explain the modest predictive performance of published models that use answers to household surveys as predictors. One feasible alternative is to increase the sampling period so that household features would arise as significant predictors of long-term airborne endotoxin levels. PMID:27065310

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

    Directory of Open Access Journals (Sweden)

    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.

  5. Endotoxin exposure and changes in short-term pulmonary function among sewage workers

    Directory of Open Access Journals (Sweden)

    Marcin Cyprowski

    2015-10-01

    Full Text Available Objectives: The inhaled endotoxin is considered as a causative factor in the process of acute bronchial obstruction, which can be measured by a decrease in forced expiratory volume in 1 s (FEV1. The aim of this study was to assess endotoxin exposure among sewage treatment plant workers (STPW and its effect on across-shift changes in respiratory airflow. Material and Methods: A group of 78 STPW from a large sewage treatment plant was studied. Inhalable dust for endotoxin assessment was collected using personal aerosol samplers. Endotoxin was assayed with the kinetic, chromogenic Limulus amebocyte lysate test. Across-shift spirometric measurements were performed on Mondays, after 2-days absence from work, with the use of portable spirometer. The forced vital capacity (FVC, and FEV1 parameters were analyzed. Multifactor regression modeling was performed to determine parameters significantly associated with endotoxin exposure. Results: The concentration of inhalable dust and endotoxin ranged from 0.01–1.38 mg/m3 and 0.68–214 endotoxin units per cubic meter of air (EU/m3, respectively. Endotoxins were characterized with the skewed distribution (arithmetic mean (AM = 38.8 EU/m3, geometric mean (GM = 15.4 EU/m3, geometric standard deviation (GSD = 4.21. Through the use of multifactor analysis, which excluded the main confounders (inhalable dust and smoking habit it was found that, despite low levels of endotoxin, it had significant impact on the observed across-shift decline in FEV1 (p = 0.044. For this parameter, the regression slope was additionally calculated (r = –0.017, p = 0.071. Conclusions: Relatively low levels of endotoxin among sewage treatment plant workers may cause small, but significant across-shift declines in FEV1. The observed relationship was independent of organic dust concentrations and smoking habit. The respiratory protection should be provided for STPW.

  6. Detection of endotoxin using a photonic crystal nanolaser

    Energy Technology Data Exchange (ETDEWEB)

    Takahashi, Daichi; Hachuda, Shoji; Watanabe, Takumi; Nishijima, Yoshiaki; Baba, Toshihiko, E-mail: baba@ynu.ac.jp [Department of Electrical and Computer Engineering, Yokohama National University, 79-5 Tokiwadai, Hodogaya-ku, Yokohama 240-8501 (Japan)

    2015-03-30

    Fast and reliable detection of endotoxin (ET) in medical equipment and pharmaceutical products is an essential precursor to clinical treatment. In this study, we demonstrate the use of shifts in wavelength of photonic crystal nanolasers for sensing the Limulus amebocyte lysate reaction, which is a standard method for detecting ET. From working curves of wavelength shift vs ET concentration, whose correlation factors were as high as 98%, we detected a required concentration of 0.001 EU/ml within 33 min and detected a low concentration of 0.0001 EU/ml.

  7. 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

  8. Airborne peptidoglycans as a supporting indicator of bacterial contamination in a metal processing plant

    Directory of Open Access Journals (Sweden)

    Marcin Cyprowski

    2016-06-01

    Full Text Available Objectives: The aim of this study was to assess exposure to airborne endotoxins and peptidoglycans (PGs as well as possibility of using PGs as a surrogate measure of bacterial exposure in workplaces in a metal processing plant. Material and Methods: Personal dosimetry (N = 11 was used to obtain data on concentrations of viable bacteria, total number of bioaerosol particles, endotoxins and peptidoglycans. To investigate the size distributions of aerosol particles responsible for transport of endotoxins and PGs, air samples (N = 5 were additionally collected using the 8-stage cascade impactor. Endotoxins and PGs were assayed with the Limulus amebocyte lysate (LAL test and a kinetic version of the silkworm larvae plasma (SLP test, respectively. Results: Median concentrations of airborne PGs (14.6 ng/m3, endotoxins (0.2 ng/m3, viable bacteria (1.16×103 CFU/m3 and the total number of bioaerosol particles (1.81×106 cells/m3 were determined. Qualitative analysis revealed presence of 19 bacterial species belonging to 14 genera. The calculations showed strong, significant correlations (p < 0.05 between endotoxins, viable bacteria (r = 0.75 and the total number of bioaerosol particle concentrations (r = 0.76 as well as between PGs and the total number of bioaerosol particle concentrations (r = 0.72. Size distribution analysis showed that the highest concentrations of bacterial aerosols occurred in the range of 2.1–3.3 μm. In the case of endotoxins, an increase of concentrations in 2 ranges of aerodynamic diameters: 1.1–3.3 μm and 5.8–9 μm was shown. For PGs there was a visible gradual increase of their concentrations in the range 2.1–9 μm. Conclusions: Peptidoglycans can be treated as a supporting indicator of bacterial contamination in metal processing plants, particularly when an assessment of an immunotoxic potential of microbiological hazards needs to be performed. However, to be extrapolated to other occupational and non

  9. ENDOTOXIN ADSORPTION IN TREATMENT OF SELECTIVE SEPTIC COMPLICATIONS IN PATIENTS WITH UROLOGICAL DISEASES AFTER RENAL TRANSPLANTATION

    Directory of Open Access Journals (Sweden)

    M. Krstic

    2014-01-01

    Full Text Available Aim. Treatment of urosepsis in patients after renal transplantation is very difficult. Currently selective endotoxin adsorption is a perspective method of treatment. Works devoted to the study of the effectiveness of this method in patients with renal transplant are extremely rare. Materials and methods. 94 recipients were included in study: 54 in prospective main group and 40 – in retrospective comparison group. For each patient of the main group we performed 2 sessions of selective endotoxin adsorption. Patients of comparison group were not trea- ted with sorption of endotoxin. We investigated the dynamics of APACHE II score, the concentration of IL-6, IL-8, TNF-alpha, procalcitonin and endotoxin. Results. As a result of endotoxin adsorption there was noted more pronounced decrease in the APACHE II score in patients of the main group, than in the comparison group. There was a decrease in activity of the systemic inflammatory response, and that was reflected in the decrease in the concentration of proinflammatory cytokines and procalcitonin in patients of the main group. Application of endotoxin adsorption resulted in a significant increase in survival in patients of the main group. Conclusion. Sorption of endotoxin is very effective and safe procedure that affects the pathogenesis of sepsis. Application of this procedure can significantly improve the condition of patients and survival. 

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

    Directory of Open Access Journals (Sweden)

    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.

  11. Selective detection of endotoxin using an impedance aptasensor with electrochemically deposited gold nanoparticles.

    Science.gov (United States)

    Su, Wenqiong; Kim, Sung-Eun; Cho, MiSuk; Nam, Jae-Do; Choe, Woo-Seok; Lee, Youngkwan

    2013-01-01

    Using a single-stranded DNA (ssDNA) aptamer exhibiting high binding affinity (Kd = 12 nM) to endotoxin as a probe, an impedance sensor where aptamer-conjugated gold nanoparticles (AuNPs) were electrochemically deposited on a gold electrode was fabricated and its performance in regard to endotoxin detection assessed. AuNPs have been employed widely as biosensors because of their unique physical and chemical properties. In order to maximize the performance of the impedance aptasensor on endotoxin detection, some critical factors affecting aptamer conjugation to AuNPs and target recognition ability (i.e. concentrations of aptamer coupled with AuNPs, pH, ion strength and cation effect at the time of aptamer-endotoxin interaction) were optimized. Electrochemical impendence spectroscopy, cyclic voltametry, atomic force microscope, scanning electron microscope and quartz crystal microbalance were employed to characterize all the modification/detection procedures during the sensor fabrication. The developed aptasensor showed a broad linear dynamic detection range (0.01-10.24 ng/ml) with a very low detection limit for endotoxin (0.005 ng/ml), despite the presence of several biomolecules (e.g. plasmid DNA, RNA, serum albumin, Glc and sucrose) known to interfere with other endotoxin assays. The demonstrated aptasensor required a detection time of only 10 min, providing a simple and fast analytical method to specifically detect endotoxin from complex biological liqors. PMID:23165992

  12. 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. PMID:25155060

  13. Dietary oil composition differentially modulates intestinal endotoxin transport and postprandial endotoxemia

    Directory of Open Access Journals (Sweden)

    Mani Venkatesh

    2013-01-01

    Full Text Available Abstract Background Intestinal derived endotoxin and the subsequent endotoxemia can be considered major predisposing factors for diseases such as atherosclerosis, sepsis, obesity and diabetes. Dietary fat has been shown to increase postprandial endotoxemia. Therefore, the aim of this study was to assess the effects of different dietary oils on intestinal endotoxin transport and postprandial endotoxemia using swine as a model. We hypothesized that oils rich in saturated fatty acids (SFA would augment, while oils rich in n-3 polyunsaturated fatty acids (PUFA would attenuate intestinal endotoxin transport and circulating concentrations. Methods Postprandial endotoxemia was measured in twenty four pigs following a porridge meal made with either water (Control, fish oil (FO, vegetable oil (VO or coconut oil (CO. Blood was collected at 0, 1, 2, 3 and 5 hours postprandial and measured for endotoxin. Furthermore, ex vivo ileum endotoxin transport was assessed using modified Ussing chambers and intestines were treated with either no oil or 12.5% (v/v VO, FO, cod liver oil (CLO, CO or olive oil (OO. Ex vivo mucosal to serosal endotoxin transport permeability (Papp was then measured by the addition of fluorescent labeled-lipopolysaccharide. Results Postprandial serum endotoxin concentrations were increased after a meal rich in saturated fatty acids and decreased with higher n-3 PUFA intake. Compared to the no oil control, fish oil and CLO which are rich in n-3 fatty acids reduced ex vivo endotoxin Papp by 50% (P  Conclusion Overall, these results indicate that saturated and n-3 PUFA differentially regulate intestinal epithelial endotoxin transport. This may be associated with fatty acid regulation of intestinal membrane lipid raft mediated permeability.

  14. Astrocytes Produce IL-19 in Response to Bacterial Challenge and are Sensitive to the Immunosuppressive Effects of this IL-10 Family Member

    Science.gov (United States)

    Cooley, Ian D.; Chauhan, Vinita S.; Donneyz, Miguel A.; Marriott, Ian

    2014-01-01

    There is growing appreciation that resident glial cells can initiate and/or regulate inflammation following trauma or infection in the central nervous system (CNS). We have previously demonstrated the ability of microglia and astrocytes to respond to bacterial pathogens or their products by rapid production of inflammatory mediators, followed by the production of the immunosuppressive cytokine interleukin (IL)210. IL-19, another member of the IL-10 family of cytokines, has been studied in the context of a number of inflammatory conditions in the periphery and is known to modulate immune cell activity. In the present study, we demonstrate the constitutive and/or inducible expression of IL-19 and its cognate receptor subunits, IL-19Rα and IL-19Rβ (also known as IL-20R1 and IL-20R2, and IL-20RA and IL-20RB), in mouse brain tissue, and by primary murine and human astrocytes. We also provide evidence for the presence of a novel truncated IL-19Rα transcript variant in mouse brain tissue, but not glial cells, that shows reduced expression following bacterial infection. Importantly, IL-19R functionality in GLIA is indicated by the ability of IL-19 to regulate signaling component expression in these cells. Furthermore, while IL-19 itself had no effect on glial cytokine production, IL-19 treatment of bacterially infected or Toll-like receptor ligand stimulated astrocytes significantly attenuated pro-inflammatory cytokine production. The bacterially induced production of IL-19 by these resident CNS cells, the constitutive expression of its cognate receptor subunits, and the immunomodulatory effects of this cytokine, suggest a novel mechanism by which astrocytes can regulate CNS inflammation. PMID:24677051

  15. Interactions between Autophagy and Bacterial Toxins: Targets for Therapy?

    Science.gov (United States)

    Mathieu, Jacques

    2015-08-04

    Autophagy is a physiological process involved in defense mechanisms for clearing intracellular bacteria. The autophagic pathway is finely regulated and bacterial toxins interact with this process in a complex manner. Bacterial toxins also interact significantly with many biochemical processes. Evaluations of the effects of bacterial toxins, such as endotoxins, pore-forming toxins and adenylate cyclases, on autophagy could support the development of new strategies for counteracting bacterial pathogenicity. Treatment strategies could focus on drugs that enhance autophagic processes to improve the clearance of intracellular bacteria. However, further in vivo studies are required to decipher the upregulation of autophagy and potential side effects limiting such approaches. The capacity of autophagy activation strategies to improve the outcome of antibiotic treatment should be investigated in the future.

  16. Interactions between Autophagy and Bacterial Toxins: Targets for Therapy?

    Directory of Open Access Journals (Sweden)

    Jacques Mathieu

    2015-08-01

    Full Text Available Autophagy is a physiological process involved in defense mechanisms for clearing intracellular bacteria. The autophagic pathway is finely regulated and bacterial toxins interact with this process in a complex manner. Bacterial toxins also interact significantly with many biochemical processes. Evaluations of the effects of bacterial toxins, such as endotoxins, pore-forming toxins and adenylate cyclases, on autophagy could support the development of new strategies for counteracting bacterial pathogenicity. Treatment strategies could focus on drugs that enhance autophagic processes to improve the clearance of intracellular bacteria. However, further in vivo studies are required to decipher the upregulation of autophagy and potential side effects limiting such approaches. The capacity of autophagy activation strategies to improve the outcome of antibiotic treatment should be investigated in the future.

  17. Research on securing no bacteria and nonfeverish property for disposable medical appliances. Inactivation of endotoxin by Co-60 {gamma} ray

    Energy Technology Data Exchange (ETDEWEB)

    Hosobuchi, Kazunari [Tokyo Metropolitan Isotope Research Center (Japan); Tanamoto, Kenichi; Haijima, Yuji

    1996-11-01

    The contamination by fever-causing endotoxin has become a large problem in medical treatment field. In the industry manufacturing disposable medical appliances, the method of manufacturing endotoxin-free products is an important subject, and the development of the methods of inactivating and eliminating efficiently endotoxin is desired. As a part of this development, the possibility of inactivating endotoxin with Co-60 {gamma} ray was examined. The sample was the endotoxin originated from E.Coli R3 F653 strain. For the irradiation, the Co-60 {gamma} ray irradiation apparatus of 185 T-Bq in National Institute of Hygienic Sciences was used. The measurement of the activity of endotoxin was carried out by limulus test synthetic substrate method. The activity value of the endotoxin in aqueous solution decreased logarithmically with the increasing irradiation dose, and this decreasing tendency was not affected by the initial concentration of the endotoxin. The experiment of recovering freezing-dried endotoxin from a vial is described. The results of inactivating the endotoxin in dry system by {gamma} ray are reported. (K.I.)

  18. Exposure to household endotoxin and total and allergen-specific IgE in the US population

    International Nuclear Information System (INIS)

    Background: Although endotoxin has strong pro-inflammatory properties, endotoxin-allergy relationship in adults and children have been inconsistent. Objectives: We investigated the association between household endotoxin levels and total immunoglobulin E (IgE) or specific IgE in the US general population, classified into three age ranges: children/adolescent, adults, and older adults. Methods: We analyzed the 2005–2006 National Health and Nutrition Examination Surveys. A total of 5220 participants for whom serum IgE and household endotoxin data were available was included in the analyses. Results: Exposure to endotoxin reduced the risk for allergic sensitization, especially in specific IgE to plants (OR in Quartile 3 = 0.58; 95% CI = 0.44–0.76) and pets (OR in Quartile 3 = 0.62; 95% CI = 0.41–0.92), for children/adolescents. In contrast, the risk among adults and older adults increased with increasing endotoxin levels. Conclusions: Our findings suggest that the effect of endotoxin on allergic reaction is likely to depend on age. - Highlights: • Findings regarding the endotoxin-allergy relationship in adults and children are inconsistent. • We investigated the association of endotoxin with total and specific IgE in US population. • The association between endotoxin levels and allergic markers is likely to depend on age. • Exposure to endotoxin reduced the risk for allergic sensitization for children/adolescents. • The risk among adults and older adults increased with increasing endotoxin levels. - Exposure to endotoxin reduced the risk for allergic sensitization for children/adolescents, but decreased the risk among adults and older

  19. The effect of endotoxin administration on the secretory dynamics of oxytocin in follicular phase mares: relationship to stress axis hormones.

    Science.gov (United States)

    Alexander, S L; Irvine, C H G

    2002-07-01

    The primary aim of this study was to define the secretory dynamics of oxytocin and vasopressin in pituitary venous effluent from ambulatory horses during acute endotoxaemia, a stimulus that may release both hormones. Our secondary aim was to investigate the role of oxytocin in regulating adrenocorticotropic hormone (ACTH) secretion by comparing oxytocin, vasopressin, corticotropin-releasing hormone (CRH) and ACTH secretory profiles during endotoxaemia and by monitoring the ACTH response to oxytocin administration. Pituitary venous blood was collected nonsurgically continuously and divided into 1-min segments from eight follicular phase mares. Four mares were sampled for 30 min before and 3.5 h after receiving an i.v. infusion of bacterial endotoxin (TOX). Four control mares were sampled for 2.5 h without infusion of TOX. Another three follicular phase mares were given 5 U of oxytocin to replicate the peak response to TOX and pituitary blood collected every 1 min for 10 min before and 15 min after injection. Endotoxin raised the secretion rates of all hormones measured. All hormones were released episodically throughout the experiment, with TOX increasing the amplitude of peaks in each hormone. Peaks in oxytocin and vasopressin were coincident in each treated mare. Similarly, ACTH peaks were coincident with peaks of oxytocin and vasopressin in each treated mare, and with peaks of CRH in three mares. However, oxytocin administration did not affect ACTH secretion. We conclude that during endotoxaemia in horses: (i) oxytocin and vasopressin are secreted synchronously; (ii) oxytocin is unlikely to be acting as an ACTH secretagogue since inducing peak oxytocin concentrations observed during TOX does not raise ACTH; and therefore (iii) the close relationship between oxytocin and ACTH secretion is circumstantial and due to the fact that oxytocin secretion is concurrent with that of vasopressin, a proven ACTH secretagogue in horses. PMID:12121490

  20. 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.

  1. 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.

  2. Altered hepatic vasopressin and alpha 1-adrenergic receptors after chronic endotoxin infusion

    Energy Technology Data Exchange (ETDEWEB)

    Roth, B.L.; Spitzer, J.A.

    1987-05-01

    Sepsis and septic shock are complicated by a number of hemodynamic and metabolic aberrations. These include catecholamine refractoriness and altered glucose metabolism. Recently, a nonshock rat model of continuous endotoxin infusion via an implanted osmotic pump was developed that reproduces some of the metabolic and cardiovascular findings of human sepsis. By using this model, we have found a decreased number of hepatic plasma membrane alpha 1-adrenergic and (Arg8)vasopressin receptors in rats continuously infused with endotoxin. There was a significant decrease in (/sup 3/H)prazosin (35 +/- 7%) and (/sup 3/H) (Arg8)vasopressin (43 +/- 8%) receptors after 30 h of continuous endotoxin infusion with no change in affinity. The ability of norepinephrine to form the high-affinity complex with alpha 1-adrenergic receptors was not altered after chronic endotoxin infusion. The results are consistent with the concept that alterations in receptor number might underlie certain of the metabolic consequences of chronic sepsis.

  3. Sensitisation to common allergens and respiratory symptoms in endotoxin exposed workers

    DEFF Research Database (Denmark)

    Basinas, Ioannis; Schlünssen, Vivi; Heederik, Dick;

    2012-01-01

    four epidemiological studies including 3883 Dutch and Danish employees in veterinary medicine, agriculture and power plants using biofuel. Endotoxin exposure was estimated by quantitative job-exposure matrices specific for the study populations. Dose–response relationships between exposure, Ig...

  4. Neural substrate of cold-seeking behavior in endotoxin shock.

    Directory of Open Access Journals (Sweden)

    Maria C Almeida

    Full Text Available Systemic inflammation is a leading cause of hospital death. Mild systemic inflammation is accompanied by warmth-seeking behavior (and fever, whereas severe inflammation is associated with cold-seeking behavior (and hypothermia. Both behaviors are adaptive. Which brain structures mediate which behavior is unknown. The involvement of hypothalamic structures, namely, the preoptic area (POA, paraventricular nucleus (PVH, or dorsomedial nucleus (DMH, in thermoregulatory behaviors associated with endotoxin (lipopolysaccharide [LPS]-induced systemic inflammation was studied in rats. The rats were allowed to select their thermal environment by freely moving in a thermogradient apparatus. A low intravenous dose of Escherichia coli LPS (10 microg/kg caused warmth-seeking behavior, whereas a high, shock-inducing dose (5,000 microg/kg caused cold-seeking behavior. Bilateral electrocoagulation of the PVH or DMH, but not of the POA, prevented this cold-seeking response. Lesioning the DMH with ibotenic acid, an excitotoxin that destroys neuronal bodies but spares fibers of passage, also prevented LPS-induced cold-seeking behavior; lesioning the PVH with ibotenate did not affect it. Lesion of no structure affected cold-seeking behavior induced by heat exposure or by pharmacological stimulation of the transient receptor potential (TRP vanilloid-1 channel ("warmth receptor". Nor did any lesion affect warmth-seeking behavior induced by a low dose of LPS, cold exposure, or pharmacological stimulation of the TRP melastatin-8 ("cold receptor". We conclude that LPS-induced cold-seeking response is mediated by neuronal bodies located in the DMH and neural fibers passing through the PVH. These are the first two landmarks on the map of the circuitry of cold-seeking behavior associated with endotoxin shock.

  5. Endotoxin concentration in poultry houses for laying hens kept in cages or in alternative housing systems

    OpenAIRE

    Huneau-Salaün, Adeline; Le Bouquin, Sophie; Bex-Capelle, Valérie; Huonnic, Didier; Balaine, Loïc; Guillam, Marie-Thérèse; Squizani, Fabrice; Ségala, Claire; Michel, Virginie

    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...

  6. Influence of various dust sampling and extraction methods on the measurement of airborne endotoxin.

    OpenAIRE

    Douwes, J; Versloot, P.; Hollander, A; Heederik, D; Doekes, G.

    1995-01-01

    The influence of various filter types and extraction conditions on the quantitation of airborne endotoxin with the Limulus amebocyte lysate test was studied by using airborne dusts sampled in a potato processing plant. Samples were collected with an apparatus designed to provide parallel samples. Data from the parallel-sampling experiment were statistically evaluated by using analysis of variance. In addition, the influence of storage conditions on the detectable endotoxin concentration was i...

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

    OpenAIRE

    Hagman R; Kindahl H; Lagerstedt A-S

    2006-01-01

    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,...

  8. Role of Thromboxane and Prostacyclin in Pulmonary Vasomotor Changes after Endotoxin in Dogs

    OpenAIRE

    Hales, C. A.; Sonne, L.; Peterson, M; Kong, D; Miller, M; Watkins, W. David

    1981-01-01

    Cyclooxygenase inhibitors prevent the pulmonary vasomotor changes in response to low-dose endotoxin. We, therefore, explored the role of two highly vasoactive prostanoids, thromboxane A2, a vasoconstrictor, and prostacyclin, a vasodilator, in the transient pulmonary vasoconstriction and subsequent loss of alveolar hypoxis vasoconstriction (AHPV) that follows endotoxin. AHPV was tested in the dog with a double-lumened endotracheal tube allowing ventilation of one lung with nitrogen as a hypoxi...

  9. Removal of Endotoxin from Human Serum Albumin Solutions by Hydrophobic and Cationic Charged Membrane

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    A novel matrix of macropore cellulose membrane was prepared by chemical graft, and immobilized the cationic charged groups as affinity ligands. The prepared membrane can be used for the removal of endotoxin from human serum albumin (HSA) solutions. With a cartridge of 20 sheets affinity membrane of 47 mm diameter, the endotoxin level in HSA solution can be reduced to 0.027 eu/mL. Recovery of HSA was over 95%.

  10. Exposure to inhalable dust and endotoxin among Danish livestock farmers: results from the SUS cohort study

    OpenAIRE

    Basinas, I.; Sigsgaard, T.; Heederik, D; Takai, H.; Omland, O; Andersen, N. T.; Wouters, I.M.; Bonlokke, J.H.; Kromhout, H; V. Schlünssen

    2012-01-01

    Studies on personal dust and endotoxin concentrations among animal farmers have been either small or limited to a few sectors in their investigations. The present study aimed to provide comparable information on the levels and variability of exposure to personal dust and endotoxin in different types of animal farmers. 507 personal inhalable dust samples were collected from 327 farmers employed in 54 pig, 26 dairy, 3 poultry, and 3 mink farms in Denmark. Measurements in pig and dairy farmers w...

  11. Comparison of Different Irrigants in the Removal of Endotoxins and Cultivable Microorganisms from Infected Root Canals

    OpenAIRE

    Marcia Carneiro Valera; Flávia Goulart da Rosa Cardoso; Adriana Chung; Ana Cláudia Carvalho Xavier; Mariana Diehl Figueiredo; Frederico Canato Martinho; Renato Miotto Palo

    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 r...

  12. Morphological characteristics of free radical processes in endotoxin damage of liver

    Directory of Open Access Journals (Sweden)

    Babanin A.A.

    2009-01-01

    Full Text Available One of the most important problems of the modern medicine is the damage of liver due to alcoholic disease. Our studies showed some features of morphological changes of liver tissues and level of biochemical markers of oxidative stress, due to experimental endotoxin intoxication. Investigations were done with the aim of modeling the endotoxin damage to the liver of rats. Investigation was carried out on 40 nonlinear rats, divided into 2 groups with 20 rats in each group. The animals of the first group were given for 7 days with intraperitoneal injections of Escherichia coli endotoxin (Sigma USA DO-235, dosage 0,1 mg/kg of body weight. Animals of the second group were used as a control. Character and level of liver damage was studied with the help of morphological methods, including electron and light microscopy. In studying of oxidative stress biochemical methods were used to define conjugated diens and dienketones, spontaneous oxidative modifications of proteins. The following results were revealed: 1 level and depth of morphological changes under endotoxin intoxication correlated with dynamic of content of products of lipid (conjugated diens and dienketones, malonic aldehyde and protein. The tendency was shown to more significant increase of oxidative modification of proteins in blood serum. It corroborated the data about primary damage of proteins under the effect of free radicals. 2 Under the influence of endotoxins of intestinal microflora, severe discirculatory changes, fat and hydropic dystrophy of hepatocytes with symptoms of toxic damage of their nucleus were found, meanwhile increased functional activity of sinusoidal cells was observed. These changes happened due to direct effects of the endotoxin, and also as a result of oxidative stress. Powerful inductor of these changes was endotoxin. Obtained results showed, that endotoxin is the main factor, which leads to worsening of vital activity of the liver.

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    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.

  15. Residual endotoxin contaminations in recombinant proteins are sufficient to activate human CD1c+ dendritic cells.

    Directory of Open Access Journals (Sweden)

    Harald Schwarz

    Full Text Available Many commercially available recombinant proteins are produced in Escherichia coli, and most suppliers guarantee contamination levels of less than 1 endotoxin unit (EU. When we analysed commercially available proteins for their endotoxin content, we found contamination levels in the same range as generally stated in the data sheets, but also some that were higher. To analyse whether these low levels of contamination have an effect on immune cells, we stimulated the monocytic cell line THP-1, primary human monocytes, in vitro differentiated human monocyte-derived dendritic cells, and primary human CD1c+ dendritic cells (DCs with very low concentrations of lipopolysaccharide (LPS; ranging from 0.002-2 ng/ml. We show that CD1c+ DCs especially can be activated by minimal amounts of LPS, equivalent to the levels of endotoxin contamination we detected in some commercially available proteins. Notably, the enhanced endotoxin sensitivity of CD1c+ DCs was closely correlated with high CD14 expression levels observed in CD1c+ DCs that had been maintained in cell culture medium for 24 hours. When working with cells that are particularly sensitive to LPS, even low endotoxin contamination may generate erroneous data. We therefore recommend that recombinant proteins be thoroughly screened for endotoxin contamination using the limulus amebocyte lysate test, fluorescence-based assays, or a luciferase based NF-κB reporter assay involving highly LPS-sensitive cells overexpressing TLR4, MD-2 and CD14.

  16. A MyD88-dependent IFNγR-CCR2 signaling circuit is required for mobilization of monocytes and host defense against systemic bacterial challenge

    Institute of Scientific and Technical Information of China (English)

    Eric M Pietras; Lloyd S Miller; Carl T Johnson; Ryan M O'Connell; Paul W Dempsey; Genhong Cheng

    2011-01-01

    Monocytes are mobilized to sites of infection via interaction between the chemokine MCP-1 and its receptor, CCR2, at which point they differentiate into macrophages that mediate potent antimicrobial effects. In this study, we investigated the mechanisms by which monocytes are mobilized in response to systemic challenge with the intracellular bacterium Francisella tularensis. We found that mice deficient in MyD88, interferon-γ (IFNγ)R or CCR2 all had defects in the expansion of splenic monocyte populations upon F. tularensis challenge, and in control of F. tularensis infection. Interestingly, MyD88-deficient mice were defective in production of IFNγ, and IFNγR deficient mice exhibited defective production of MCP-1, the ligand for CCR2. Transplantation of IFNγR-deficient bone marrow (BM) into wild-type mice further suggested that mobilization of monocytes in response to F. tularensis challenge required IFNγR expression on BM-derived cells. These studies define a critical host defense circuit wherein MyD88-dependent IFNγ production signals via IFNγR expressed on BM-derived cells, resulting in MCP-1 production and activation of CCR2-dependent mobilization of monocytes in the innate immune response to systemic F. tularensis challenge.

  17. Effects of a dietary yeast extract on hematological parameters, heterophil function, and bacterial clearance in turkey poults challenged with Escherichia coli and subjected to transport stress

    Science.gov (United States)

    There is a need to develop nutritional methods for controlling pathogens in poultry production. A standardized yeast extract supplement, Alphamune™ (YE), was added to turkey poult diets. Male poults were challenged by air sac injection with 60 cfu of E. coli at 1 week of age. At 3 weeks of age chal...

  18. Detection of endotoxins in radiopharmaceutical preparations. II. Comparison of the sensitivity of methods using the rabbit and the Limulus amoebocyte lysate for the detection of endotoxins

    Energy Technology Data Exchange (ETDEWEB)

    Bruneau, J.; Cohen, Y.; Merlin, L.; Peysson, S.

    1986-01-01

    The rise of the rabbit internal temperature after i.v. injection of an endotoxin solution is proportional to concentration. Gelation of Limulus amoebocyte, when in presence of an endotoxin solution, is also related to concentration. We compared the sensitivity of these two methods. With our experimental procedure, the rabbit is sensitive to a 0.40 ng/mL solution and the Limulus amoebocyte lysate to a 0.14 ng/mL solution. The rabbit sensitivity increase is related to the per kilogramme injected volume, whereas sensitivity is not related to the volume to check in the case of the lysate.

  19. Dose-dependent effects of endotoxin on neurobehavioral functions in humans.

    Directory of Open Access Journals (Sweden)

    Jan-Sebastian Grigoleit

    Full Text Available Clinical and experimental evidence document that inflammation and increased peripheral cytokine levels are associated with depression-like symptoms and neuropsychological disturbances in humans. However, it remains unclear whether and to what extent cognitive functions like memory and attention are affected by and related to the dose of the inflammatory stimulus. Thus, in a cross-over, double-blind, experimental approach, healthy male volunteers were administered with either placebo or bacterial lipopolysaccharide (LPS at doses of 0.4 (n = 18 or 0.8 ng/kg of body weight (n = 16. Pro- and anti-inflammatory cytokines, norephinephrine and cortisol concentrations were analyzed before and 1, 1.75, 3, 4, 6, and 24 h after injection. In addition, changes in mood and anxiety levels were determined together with working memory (n-back task and long term memory performance (recall of emotional and neutral pictures of the International Affective Picture System. Endotoxin administration caused a profound transient physiological response with dose-related elevations in body temperature and heart rate, increases in plasma interleukin (IL-6, IL-10, tumor necrosis factor (TNF-α and IL-1 receptor antagonist (IL-1ra, salivary and plasma cortisol, and plasma norepinephrine. These changes were accompanied by dose-related decreased mood and increased anxiety levels. LPS administration did not affect accuracy in working memory performance but improved reaction time in the high-dose LPS condition compared to the control conditon. In contrast, long-term memory performance was impaired selectively for emotional stimuli after administration of the lower but not of the higher dose of LPS. These data suggest the existence of at least two counter-acting mechanisms, one promoting and one inhibiting cognitive performance during acute systemic inflammation.

  20. Relationship between endotoxin and prostaglandin (PGE2 and PGFM) concentrations and ovarian function in dairy cows with puerperal endometritis.

    Science.gov (United States)

    Mateus, L; Lopes da Costa, L; Diniz, P; Ziecik, A J

    2003-04-15

    Blood concentrations of progesterone, 13,14-dihydro-15-keto-prostaglandin F2alpha (PGFM) and endotoxin, and uterine fluid concentrations of prostaglandin E(2) (PGE(2)), PGFM and endotoxin were evaluated in 14 dairy cows with puerperal endometritis (mild (n=6) and heavy (n=8)). Endotoxin was measured using a quantitative kinetic assay. Cows with heavy endometritis had significantly higher concentrations of plasma PGFM (P<0.01) and uterine fluid PGE(2) and endotoxin (P<0.05) than cows with mild endometritis. Concentrations of PGFM in plasma and uterine fluid, of PGFM and PGE(2), and PGE(2) and endotoxin in uterine fluid were positively and significantly (P<0.05) correlated. The presence of endotoxin in plasma was detected in one out of six mild and in eight out of eight heavy endometritis cows. Peak plasma endotoxin concentrations (0.08-9.14 endotoxin units/ml (EU/ml) were observed between 1 and 12 days postpartum (pp) and thereafter amounts generally remained below 0.1 EU/ml (last day of detection: Day 27 pp). Abnormal ovarian function was observed in six cows (four with prolonged anoestrus and two with long luteal phase after the first postpartum ovulation). Plasma endotoxin concentrations were detected in the anoestric cows. The results suggest that: (i) concentrations of uterine fluid endotoxin and PGE(2) and of plasma PGFM are related to the degree of endometritis; (ii) absorption of endotoxin from the uterus to the bloodstream occurs, mainly in heavy endometritis cows; and (iii) there is a relationship between uterine infection, endotoxin production and resumption of pp ovarian activity. PMID:12586488

  1. Evaluation on Sensitivity of the Human Sperm Motility Assay for Detecting Endotoxin in Culture Medium

    Institute of Scientific and Technical Information of China (English)

    Wei-jie ZHU; Jing LI; Wen-hong ZHANG; Kang-shou YAO

    2003-01-01

    Objective To investigate the sensitivity of the human sperm motility assay for detecting endotoxin in culture medium Materials & Methods Motile sperm were separated and exposed to different concentrations of endotoxin (0.5 ng/mL, 1 ng/mL, 10 ng/mL, 1 000 ng/mL, 10 000 ng/mL, and 50 000 ng/mL), and sperm motility was determined after incubation. Effects of endotoxin on sperm motility in media without albumin were also examined. In addition, at the same concentrations of endotoxin (0.5 ng/mL, 1 ng/mL, and 10 ng/mL), the sensitivity of the human sperm motility assay was compared to those of 1-cell and 2-cell mouse embryo bioassays.Results At levels of 0.5 ng/mL~1 000 ng/mL endotoxin in media with 2 mg/mL albumin, sperm did not show significant change in motility during 24 h of incubation when compared with the control (P>0.05). However, the sperm motility was significantly inhibited at endotoxin dosages of 10 000 and 50 000 ng/mL. In the absence of albumin supplementation, at endotoxin levels of 50 000 ng/mL, and 1 000 ng/mL, there was a marked decrease in sperm motility compared with the control after 2 h or 8 h of incubation, respectively (P<0.01). In media containing 0.5 ng/mL and 1 ng/mL endotoxin, 1-cell and 2-cell mouse embryos had significantly reduced developmental rates in all developmental stages, and at the level of 10 ng/mL, the development of the embryos was arrested.Conclusion The human sperm motility assay could detect high levels of endotoxin in culture medium but its sensitivity to endotoxin would be inferior to that of the 1-cell or 2-cell mouse embryo bioassay. In the absence of albumin supplementation, the sensitivity of the sperm motility assay could be improved.

  2. A practical approach for the validation of sterility, endotoxin and potency testing of bone marrow mononucleated cells used in cardiac regeneration in compliance with good manufacturing practice

    Directory of Open Access Journals (Sweden)

    Gola Mauro

    2009-09-01

    Full Text Available Abstract Background Main scope of the EU and FDA regulations is to establish a classification criterion for advanced therapy medicinal products (ATMP. Regulations require that ATMPs must be prepared under good manufacturing practice (GMP. We have validated a commercial system for the determination of bacterial endotoxins in compliance with EU Pharmacopoeia 2.6.14, the sterility testing in compliance with EU Pharmacopoeia 2.6.1 and a potency assay in an ATMP constituted of mononucleated cells used in cardiac regeneration. Methods For the potency assay, cells were placed in the upper part of a modified Boyden chamber containing Endocult Basal Medium with supplements and transmigrated cells were scored. The invasion index was expressed as the ratio between the numbers of invading cells relative to cell migration through a control insert membrane. For endotoxins, we used a commercially available system based on the kinetic chromogenic LAL-test. Validation of sterility was performed by direct inoculation of TSB and FTM media with the cell product following Eu Ph 2.6.1 guideline. Results and discussion The calculated MVD and endotoxin limit were 780× and 39 EU/ml respectively. The 1:10 and 1:100 dilutions were selected for the validation. For sterility, all the FTM cultures were positive after 3 days. For TSB cultures, Mycetes and B. subtilis were positive after 5 and 3 days respectively. The detection limit was 1-10 colonies. A total of four invasion assay were performed: the calculated invasion index was 28.89 ± 16.82% (mean ± SD. Conclusion We have validated a strategy for endotoxin, sterility and potency testing in an ATMP used in cardiac regeneration. Unlike pharmaceutical products, many stem-cell-based products may originate in hospitals where personnel are unfamiliar with the applicable regulations. As new ATMPs are developed, the regulatory framework is likely to evolve. Meanwhile, existing regulations provide an appropriate structure for

  3. 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

    accumulation of platelets in pulmonary capillaries initiating an acute and chronic inflammation resulting in endothelial cell damage and extravasation of cells and fluids into the lung interstitium. These changes cause bronchoconstriction, the decrement of lung function expressed as reduction of forced expiratory volume in one second (FEV1) and/or diffusion capacity, increase in the airway hyperreactivity and subjective symptoms such as fever, airway irritation and chest tightness. The conclusions from these experiments, performed mostly 2-3 decades ago, did not loose their actuality until recently as so far no other cotton dust component was identified as a more important work-related hazard than bacterial endotoxin. Though also other microbial and plant constituents are considered as potential causative agents of byssinosis, the endotoxin produced by Pantoea agglomerans and other Gram-negative bacteria present in cotton dust is still regarded as a major cause of this mysterious disease. PMID:26706959

  4. 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

    that cause accumulation of platelets in pulmonary capillaries initiating an acute and chronic inflammation resulting in endothelial cell damage and extravasation of cells and fluids into the lung interstitium. These changes cause bronchoconstriction, the decrement of lung function expressed as reduction of forced expiratory volume in one second (FEV1 and/or diffusion capacity, increase in the airway hyperreactivity and subjective symptoms such as fever, airway irritation and chest tightness. The conclusions from these experiments, performed mostly 2-3 decades ago, did not loose their actuality until recently as so far no other cotton dust component was identified as a more important work-related hazard than bacterial endotoxin. Though also other microbial and plant constituents are considered as potential causative agents of byssinosis, the endotoxin produced by Pantoea agglomerans and other Gram-negative bacteria present in cotton dust is still regarded as a major cause of this mysterious disease.

  5. Responding to the challenge of untreatable gonorrhea: ETX0914, a first-in-class agent with a distinct mechanism-of-action against bacterial Type II topoisomerases.

    Science.gov (United States)

    Basarab, Gregory S; Kern, Gunther H; McNulty, John; Mueller, John P; Lawrence, Kenneth; Vishwanathan, Karthick; Alm, Richard A; Barvian, Kevin; Doig, Peter; Galullo, Vincent; Gardner, Humphrey; Gowravaram, Madhusudhan; Huband, Michael; Kimzey, Amy; Morningstar, Marshall; Kutschke, Amy; Lahiri, Sushmita D; Perros, Manos; Singh, Renu; Schuck, Virna J A; Tommasi, Ruben; Walkup, Grant; Newman, Joseph V

    2015-01-01

    With the diminishing effectiveness of current antibacterial therapies, it is critically important to discover agents that operate by a mechanism that circumvents existing resistance. ETX0914, the first of a new class of antibacterial agent targeted for the treatment of gonorrhea, operates by a novel mode-of-inhibition against bacterial type II topoisomerases. Incorporating an oxazolidinone on the scaffold mitigated toxicological issues often seen with topoisomerase inhibitors. Organisms resistant to other topoisomerase inhibitors were not cross-resistant with ETX0914 nor were spontaneous resistant mutants to ETX0914 cross-resistant with other topoisomerase inhibitor classes, including the widely used fluoroquinolone class. Preclinical evaluation of ETX0914 pharmacokinetics and pharmacodynamics showed distribution into vascular tissues and efficacy in a murine Staphylococcus aureus infection model that served as a surrogate for predicting efficacious exposures for the treatment of Neisseria gonorrhoeae infections. A wide safety margin to the efficacious exposure in toxicological evaluations supported progression to Phase 1. Dosing ETX0914 in human volunteers showed sufficient exposure and minimal adverse effects to expect a highly efficacious anti-gonorrhea therapy. PMID:26168713

  6. Bacterial modulation of the lung response to coal mine dust

    Energy Technology Data Exchange (ETDEWEB)

    Cullen, R.T.; Slight, J.; MacLaren, W.M. [Institute of Occupational Medicine, Edinburgh (United Kingdom)

    1997-12-31

    A study is reported which examined, in an animal model, the effects of bacteria (Corynebacterium parvum) and a bacterial product, lipopolysaccharide (endotoxin), on the pulmonary response to instilled coal mine dust. Dust from the air of a British mine producing a low rank coal was instilled into the lungs of SPF male Wistar rats. The study concluded that infection, both in the lung and at remote sites, could modulate the activity of the alveolar macrophage, a key cell in dust pathogenesis because of its ability to imitate and regulate inflammation and fibrogenesis. 17 refs., 4 figs.

  7. IMPACT OF OBESITY ON ENDOTOXIN-INDUCED DISSEMINATED INTRAVASCULAR COAGULATION.

    Science.gov (United States)

    Duburcq, Thibault; Tournoys, Antoine; Gnemmi, Viviane; Hubert, Thomas; Gmyr, Valery; Pattou, François; Jourdain, Mercé

    2015-10-01

    An early activation of coagulation and fibrinolysis occurs during sepsis, leading to the syndrome of disseminated intravascular coagulation (DIC). Obesity has been demonstrated to be a hypercoagulable and hypofibrinolytic state, but its impact on DIC has never been studied. In this study, we aimed to determine if obesity impairs DIC in an acute endotoxic shock model using minipigs. This was a prospective, comparative, and experimental ancillary study approved by the Animal Ethics Committee. Pigs were chosen as a clinically relevant species, resembling humans in coagulation reactions. Four groups of five "Yucatan" minipigs were studied: lean and obese control groups, a lean lipopolysaccharide (LPS) group receiving Escherichia coli endotoxin (LPS), and an obese LPS group receiving the same endotoxin dose. We measured standard coagulation parameters (prothrombin time [PT], platelet count, and fibrinogen levels), thrombin-antithrombin complexes, tissue-type plasminogen activator, and plasminogen activator inhibitor-1. All measurements were performed at baseline and 30, 60, 90, 150, and 300 min. Results were given as median with interquartile ranges. At baseline, platelet count (477 [428 - 532] G/L vs. 381 [307 - 442] G/L; P = 0.005) and fibrinogen levels (4.6 [3.8 - 5.2] g/L vs. 2 [1.8 - 2.9] g/L; P coagulation parameters (PT, platelet count, and fibrinogen levels) and the increase in thrombin-antithrombin complexes (581 [382 - 1,057] μg/mL vs. 247 [125 - 369] μg/mL at 150 min; P = 0.03) were significantly more important in the obese LPS group compared with those in the lean LPS group. Concerning the fibrinolytic reaction, we found a slightly more elevated increase of plasminogen activator inhibitor-1 in the obese LPS group at 300 min (481 [365 - 617] ng/mL vs. 355 [209 - 660] ng/mL; P = 0.66). In our model of endotoxic shock, obese pigs developed a more severe DIC with a more severe procoagulant response.

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

    Directory of Open Access Journals (Sweden)

    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.

  9. MAJOR LYMPHOCYTE SUBPOPULATIONS IN PATIENTS WITH SYSTEMIC LUPUS ERYTHEMATOSUS AND THEIR ASSOCIATIONS WITH CELLULAR AND HUMORAL ANTI-ENDOTOXIN IMMUNITY

    Directory of Open Access Journals (Sweden)

    D. V. Shaduro

    2015-01-01

    Full Text Available At the present time, systemic lupus erythematosus (SLE takes the leading place among systemic autoimmune disorders. Despite considerable progress in understanding basic pathogenesis of this disease, many subtle mechanisms of progressive inflammation in SLE are still unknown. It has been discovered that the persistent self-maintenance factors of autoimmune inflammation could be represented by lipopolysaccharides or endotoxins of Gram-negative intestinal bacteria. The objective of this study was to assess the levels of major lymphocyte subpopulations, and their probable relation to specific anti-endotoxin antibodies and endotoxin-neutralizing receptors of granulocytes and monocytes in peripheral blood of SLE patients. The study involved forty-eight patients with SLE. The levels of lymphocyte subpopulations, expression of monocyte and granulocyte anti-endotoxin receptors, amounts of total and endotoxin-specific immunoglobulins were determined by means of, respectively, cytometric analysis and enzyme immunoassay techniques. The results of study have shown an increase in overall numbers of activated and cytotoxic T lymphocytes, a decrease in lymphocytes and NK-cells, diminished levels endotoxin-binding receptors on the monocytes and granulocytes, along with increased anti-endotoxin IgG antibodies. Our study revealed correlations between the levels of the leukocyte endotoxin-binding receptors, and B-lymphocyte contents, like as some associations between anti-endotoxin IgM antibodies, and the levels of B-lymphocytes, and cytotoxic T-lymphocytes. A correlation was also found between anti-endotoxin IgG antibodies and CD4+ lymphocyte levels. Significant alterations of the endotoxin-specific immunity among SLE patients suggest that this imbalance might play an important role in the mechanisms of onset and progression of autoimmune diseases.

  10. Responses of three very large inducible GTPases to bacterial and white spot syndrome virus challenges in the giant fresh water prawn Macrobrachium rosenbergii.

    Science.gov (United States)

    Huang, Ying; Jin, Min; Yin, Shaowu; Ding, Zhengfeng; Wang, Wen; Ren, Qian

    2016-04-01

    Interferons (IFNs) are cytokines secreted by cells in response to invasion by pathogens, such as viruses, bacteria, parasites, or tumor cells. Very large inducible GTPases (VLIG) are the latest IFN-inducible GTPase family to be discovered and are the largest known GTPases of any species. However, VLIG proteins from invertebrates have yet to be characterized. In this study, three forms of VLIGs designated as MrVLIG1, MrVLIG2, and MrVLIG3 were cloned from the giant fresh water prawn Macrobrachium rosenbergii. MrVLIG1 has a 5445 bp open reading frame (ORF) encoding an 1814-amino acid protein. The complete nucleotide sequence of MrVLIG2 cDNA is 7055 bp long consisting of a 5757 bp ORF encoding a protein with 1918 amino acids. The full length of the MrVLIG3 gene consists of 5511 bp with a 3909 bp ORF encoding a peptide with 1302 amino acids. BLASTP and phylogenetic tree analyses showed that the three MrVLIGs are clustered into one subgroup and, together with other vertebrate VLIGs, into a branch. Tissue distribution analysis indicated that the mRNAs of the three MrVLIGs were widely expressed in almost all detected tissues, including the hemocytes, heart, hepatopancreas, gills, stomach, and intestine, with the highest expression in the hepatopancreas. They were also detected in the intestine but with relatively low expression levels. Quantitative real-time RT-PCR analysis showed that the mRNA transcripts of the MrVLIGs in the hepatopancreas were significantly expressed at various time points after infection with Vibrio parahaemolyticus and white spot syndrome virus. In summary, the three isoforms of VLIG genes participate in the innate immune response of the shrimps to bacterial and viral infections.

  11. Responses of three very large inducible GTPases to bacterial and white spot syndrome virus challenges in the giant fresh water prawn Macrobrachium rosenbergii.

    Science.gov (United States)

    Huang, Ying; Jin, Min; Yin, Shaowu; Ding, Zhengfeng; Wang, Wen; Ren, Qian

    2016-04-01

    Interferons (IFNs) are cytokines secreted by cells in response to invasion by pathogens, such as viruses, bacteria, parasites, or tumor cells. Very large inducible GTPases (VLIG) are the latest IFN-inducible GTPase family to be discovered and are the largest known GTPases of any species. However, VLIG proteins from invertebrates have yet to be characterized. In this study, three forms of VLIGs designated as MrVLIG1, MrVLIG2, and MrVLIG3 were cloned from the giant fresh water prawn Macrobrachium rosenbergii. MrVLIG1 has a 5445 bp open reading frame (ORF) encoding an 1814-amino acid protein. The complete nucleotide sequence of MrVLIG2 cDNA is 7055 bp long consisting of a 5757 bp ORF encoding a protein with 1918 amino acids. The full length of the MrVLIG3 gene consists of 5511 bp with a 3909 bp ORF encoding a peptide with 1302 amino acids. BLASTP and phylogenetic tree analyses showed that the three MrVLIGs are clustered into one subgroup and, together with other vertebrate VLIGs, into a branch. Tissue distribution analysis indicated that the mRNAs of the three MrVLIGs were widely expressed in almost all detected tissues, including the hemocytes, heart, hepatopancreas, gills, stomach, and intestine, with the highest expression in the hepatopancreas. They were also detected in the intestine but with relatively low expression levels. Quantitative real-time RT-PCR analysis showed that the mRNA transcripts of the MrVLIGs in the hepatopancreas were significantly expressed at various time points after infection with Vibrio parahaemolyticus and white spot syndrome virus. In summary, the three isoforms of VLIG genes participate in the innate immune response of the shrimps to bacterial and viral infections. PMID:26850335

  12. Bacterial Vaginosis

    Science.gov (United States)

    ... 586. Related Content STDs during Pregnancy Fact Sheet Pregnancy and HIV, Viral Hepatitis, and STD Prevention Pelvic Inflammatory Disease ( ... Bacterial Vaginosis (BV) Chlamydia Gonorrhea Genital Herpes Hepatitis HIV/AIDS & STDs Human Papillomavirus ... STDs See Also Pregnancy Reproductive ...

  13. Bacterial Meningitis

    Science.gov (United States)

    ... Schedules Preteen & Teen Vaccines Meningococcal Disease Sepsis Bacterial Meningitis Recommend on Facebook Tweet Share Compartir On this ... serious disease. Laboratory Methods for the Diagnosis of Meningitis This manual summarizes laboratory methods used to isolate, ...

  14. CCL2 mediates the circadian response to low dose endotoxin.

    Science.gov (United States)

    Duhart, José M; Brocardo, Lucila; Mul Fedele, Malena L; Guglielmotti, Angelo; Golombek, Diego A

    2016-09-01

    The mammalian circadian system is mainly originated in a master oscillator located in the suprachiasmatic nuclei (SCN) in the hypothalamus. Previous reports from our and other groups have shown that the SCN are sensitive to systemic immune activation during the early night, through a mechanism that relies on the action of proinflammatory factors within this structure. Chemokine (C-C motif) ligand 2 (CCL2) is induced in the brain upon peripheral immune activation, and it has been shown to modulate neuronal physiology. In the present work we tested whether CCL2 might be involved in the response of the circadian clock to peripheral endotoxin administration. The CCL2 receptor, C-C chemokine receptor type 2 (CCR2), was detected in the SCN of mice, with higher levels of expression during the early night, when the clock is sensitive to immune activation. Ccl2 was induced in the SCN upon intraperitoneal lipopolysaccharide (LPS) administration. Furthermore, mice receiving an intracerebroventricular (Icv) administration of a CCL2 synthesis inhibitor (Bindarit), showed a reduction LPS-induced circadian phase changes and Icv delivery of CCL2 led to phase delays in the circadian clock. In addition, we tested the possibility that CCL2 might also be involved in the photic regulation of the clock. Icv administration of Bindarit did not modify the effects of light pulses on the circadian clock. In summary, we found that CCL2, acting at the SCN level is important for the circadian effects of immune activation. PMID:27178133

  15. In vitro assay of endotoxin by the inhibition of macrophage migration.

    Science.gov (United States)

    Heilman, D H; Bast, R C

    1967-01-01

    A quantitative in vitro technique was used to compare the ability of different endotoxins to inhibit the migration of macrophages from explants of rabbit spleen cultured in a coagulated plasma medium. The order of potency was different from that observed in chick embryo assays, and in assays with mice, of the same endotoxins. In general, however, the sensitivity of the macrophage inhibition test was comparable to that of other bioassay methods. A highly purified endotoxin from Salmonella enteritidis (Ribi) in a concentration of 0.004 mug/ml regularly inhibited macrophage migration. The in vitro method was used to detect a progressive loss of biological activity in fractions obtained during acid hydrolysis of the purified endotoxin. The selective toxicity of very low concentrations of endotoxin for mammalian macrophages was important in estimating the degree of specificity of the reaction. The pattern of cellular response in explant cultures made it possible to differentiate endotoxic damage from the specific cytotoxic action of antigen associated with delayed hypersensitivity. PMID:5335889

  16. Toll-like receptor and tumour necrosis factor dependent endotoxin-induced acute lung injury

    Science.gov (United States)

    Togbe, Dieudonnée; Schnyder-Candrian, Silvia; Schnyder, Bruno; Doz, Emilie; Noulin, Nicolas; Janot, Laure; Secher, Thomas; Gasse, Pamela; Lima, Carla; Coelho, Fernando Rodrigues; Vasseur, Virginie; Erard, François; Ryffel, Bernhard; Couillin, Isabelle; Moser, Rene

    2007-01-01

    Recent studies on endotoxin/lipopolysaccharide (LPS)-induced acute inflammatory response in the lung are reviewed. The acute airway inflammatory response to inhaled endotoxin is mediated through Toll-like receptor 4 (TLR4) and CD14 signalling as mice deficient for TLR4 or CD14 are unresponsive to endotoxin. Acute bronchoconstriction, tumour necrosis factor (TNF), interleukin (IL)-12 and keratinocyte-derived chemokine (KC) production, protein leak and neutrophil recruitment in the lung are abrogated in mice deficient for the adaptor molecules myeloid differentiation factor 88 (MyD88) and Toll/Interleukin-1 receptor (TIR)-domain-containing adaptor protein (TIRAP), but independent of TIR-domain-containing adaptor-inducing interferon-beta (TRIF). In particular, LPS-induced TNF is required for bronchoconstriction, but dispensable for inflammatory cell recruitment. Lipopolysaccharide induces activation of the p38 mitogen-activated protein kinase (MAPK). Inhibition of pulmonary MAPK activity abrogates LPS-induced TNF production, bronchoconstriction, neutrophil recruitment into the lungs and broncho-alveolar space. In conclusion, TLR4-mediated, bronchoconstriction and acute inflammatory lung pathology to inhaled endotoxin are dependent on TLR4/CD14/MD2 expression using the adapter proteins TIRAP and MyD88, while TRIF, IL-1R1 or IL-18R signalling pathways are dispensable. Further downstream in this axis of signalling, TNF blockade reduces only acute bronchoconstriction, while MAPK inhibition abrogates completely endotoxin-induced inflammation. PMID:18039275

  17. Bacillus thuringiensis endotoxins active against Chilo partellus and Glossina morsitans morsitans

    International Nuclear Information System (INIS)

    Bacillus thuringiensis crystal endotoxins were isolated by centrifugation on linear sucrose gradients. Analysis of the crystals by gel electrophoresis revealed that the major component of the Chilo partellus active crystal endotoxin was a protein of Mr ∼ 130 kilodalton. The Glossina morsitans morsitans active crystal endotoxin gave a major protein band of Mr ∼ 120 kilodalton. Upon solubilization under alkaline pH and reducing conditions, the C. partellus and G. m. morsitans crystal endotoxin yielded protoxins of Mr ∼ 63 and Mr ∼ 64 kilodalton, respectively. Activation of the C. partellus protoxin with bovine trypsin resulted in no apparent change in the molecular weight. However, treatment with bovine chymotrypsin or C. partellus midgut homogenate resulted in a shift in the molecular weight of the protoxin to a toxin of Mr ∼ 60 kilodalton. Similarly, treatment of G.m. morsitans protoxin with bovine trypsin gave a toxin of Mr ∼ 62 kilodalton, but bovine chymotrypsin gave a toxin of Mr ∼ 60 kilodalton. Staining with periodic acid Schiff reagent revealed that both the crystal endotoxins were glycosylated. The carbohydrate moieties were of the high mannose type, as shown by staining with fluorescein isothiocyanate conjugated-concanavalin A. Rabbit antibodies against C. partellus protoxin cross-reacted with the G. m. morsitans toxin. (author). 19 refs, 5 figs

  18. Effectiveness of castor oil extract on Escherichia coli and its endotoxins in root canals.

    Science.gov (United States)

    Valera, Marcia Carneiro; Maekawa, Lilian Eiko; Chung, Adriana; de Oliveira, Luciane Dias; Carvalho, Claudio Antonio Talge; Koga-Ito, Cristiane Yumi; Jorge, Antonio Olavo Cardoso

    2012-01-01

    This in vitro study sought to evaluate the effectiveness of castor oil extract used as an irrigating solution on Escherichia coli and its endotoxins in root canals. Sixty single-rooted teeth were prepared (using castor oil extract as irrigating solution) and divided into five groups (n = 12): Group 1 samples were treated with calcium hydroxide (Ca(OH)2), Group 2 samples were treated with polymyxin B, Group 3 samples were treated with Ca(OH)2 and 2% chlorhexidine gel (CHX), and Group 4 samples were treated with castor oil extract. A control group used physiological saline solution as an irrigant. Canal content samples were collected at four different times: immediately after instrumentation, seven days after instrumentation, after 14 days of intracanal medication, and seven days after removal of intracanal medication. A plating method was used to assess antimicrobial activity and the quantification of endotoxins was evaluated by the chromogenic Limulus lysate assay. Data were submitted to ANOVA and a Dunn test (a = 5%). Irrigation with castor oil extract decreased E. coli counts but had no effect on the level of endotoxins. Samples taken seven days after removal of medication revealed a significant reduction in endotoxin levels in Groups 3 and 4. Compared to the saline solution irrigation, castor oil extract decreased microorganism counts in root canals immediately after canal preparation. None of the medications used completely eliminated endotoxins in the root canal.

  19. Obesity Increases Sensitivity to Endotoxin Liver Injury: Implications for the Pathogenesis of Steatohepatitis

    Science.gov (United States)

    Yang, Shi Qi; Zhi Lin, Hui; Lane, M. Daniel; Clemens, Mark; Diehl, Anna Mae

    1997-03-01

    Genetically obese fatty/fatty rats and obese/obese mice exhibit increased sensitivity to endotoxin hepatotoxicity, quickly developing steatohepatitis after exposure to low doses of lipopolysaccharide (LPS). Among obese animals, females are more sensitive to endotoxin liver injury than males. LPS induction of tumor necrosis factor α (TNFα ), the proven affecter of endotoxin liver injury, is no greater in the livers, white adipose tissues, or sera of obese animals than in those of lean controls. Indeed, the lowest serum concentrations of TNF occur in female obese rodents, which exhibit the most endotoxin-induced liver injury. Several cytokines that modulate the biological activity of TNF are regulated abnormally in the livers of obese animals. After exposure to LPS, mRNA of interferon γ , which sensitizes hepatocytes to TNF toxicity, is overexpressed, and mRNA levels of interleukin 10, a TNF inhibitor, are decreased. The phagocytic activity of liver macrophages and the hepatic expression of a gene encoding a macrophage-specific receptor are also decreased in obesity. This new animal model of obesity-associated liver disease demonstrates that hepatic macrophage dysfunction occurs in obesity and suggests that this might promote steatohepatitis by sensitizing hepatocytes to endotoxin.

  20. Endotoxin concentration in poultry houses for laying hens kept in cages or in alternative housing systems.

    Science.gov (United States)

    Huneau-Salaün, A; Le Bouquin, S; Bex-Capelle, V; Huonnic, D; Balaine, L; Guillam, M-T; Squizani, F; Segala, C; Michel, V

    2011-10-01

    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. The endotoxin concentrations in the inhalable fraction of airborne dust (below 100 µm) from cage and alternative system houses (on-floor, free range and aviaries) were compared under both experimental and commercial conditions. The endotoxin concentration was higher in experimental aviaries (median: 565 EU/m³, range: 362-1491 EU/m³) than in cage housing (98 EU/m³ (51-470)). In field conditions, the endotoxin concentration in the air of 13 alternative houses was higher (35 to 3156 EU/m³) than in cage system buildings (n = 8, 78-576 EU/m³). It was correlated to the respirable dust concentration (fraction below 5 µm) and to the temperature inside the hen house but no seasonal variation was observed. The present study emphasises that considerable worker exposure to endotoxins may occur in laying houses, especially in alternative systems. PMID:22029777

  1. Oxidative degradation of endotoxin by advanced oxidation process (O3/H2O2 & UV/H2O2).

    Science.gov (United States)

    Oh, Byung-Taek; Seo, Young-Suk; Sudhakar, Dega; Choe, Ji-Hyun; Lee, Sang-Myeong; Park, Youn-Jong; Cho, Min

    2014-08-30

    The presence of endotoxin in water environments may pose a serious public health hazard. We investigated the effectiveness of advanced oxidative processes (AOP: O3/H2O2 and UV/H2O2) in the oxidative degradation of endotoxin. In addition, we measured the release of endotoxin from Escherichia coli following typical disinfection methods, such as chlorine, ozone alone and UV, and compared it with the use of AOPs. Finally, we tested the AOP-treated samples in their ability to induce tumor necrosis factor alpha (TNF-α) in mouse peritoneal macrophages. The production of hydroxyl radical in AOPs showed superior ability to degrade endotoxin in buffered solution, as well as water samples from Korean water treatment facilities, with the ozone/H2O2 being more efficient compared to UV/H2O2. In addition, the AOPs proved effective not only in eliminating E. coli in the samples, but also in endotoxin degradation, while the standard disinfection methods lead to the release of endotoxin following the bacteria destruction. Furthermore, in the experiments with macrophages, the AOPs-deactivated endotoxin lead to the smallest induction of TNF-α, which shows the loss of inflammation activity, compared to ozone treatment alone. In conclusion, these results suggest that AOPs offer an effective and mild method for endotoxin degradation in the water systems.

  2. Effects of endotoxin exposure on childhood asthma risk are modified by a genetic polymorphism in ACAA1

    Directory of Open Access Journals (Sweden)

    Sordillo Joanne E

    2011-12-01

    Full Text Available Abstract Background Polymorphisms in the endotoxin-mediated TLR4 pathway genes have been associated with asthma and atopy. We aimed to examine how genetic polymorphisms in innate immunity pathways interact with endotoxin to influence asthma risk in children. Methods In a previous analysis of 372 children from the Boston Home Allergens and the Connecticut Childhood Asthma studies, 7 SNPs in 6 genes (CARD15, TGFB1, LY96, ACAA1, DEFB1 and IFNG involved in innate immune pathways were associated with asthma, and 5 SNPs in 3 genes (CD80, STAT4, IRAK2 were associated with eczema. We tested these SNPs for interaction with early life endotoxin exposure (n = 291, in models for asthma and eczema by age 6. Results We found a significant interaction between endotoxin and a SNP (rs156265 in ACAA1 (p = 0.0013 for interaction. Increased endotoxin exposure (by quartile showed protective effects for asthma in individuals with at least one copy of the minor allele (OR = 0.39 per quartile increase in endotoxin, 95% CI 0.15 to 1.01. Endotoxin exposure did not reduce the risk of asthma in children homozygous for the major allele. Conclusion Our findings suggest that protective effects of endotoxin exposure on asthma may vary depending upon the presence or absence of a polymorphism in ACAA1.

  3. Seasonal variation in airborne endotoxin levels in indoor environments with different micro-environmental factors in Seoul, South Korea.

    Science.gov (United States)

    Hwang, Sung Ho; Park, Dong Jin; Park, Wha Me; Park, Dong Uk; Ahn, Jae Kyoung; Yoon, Chung Sik

    2016-02-01

    This study evaluated the variation over a year in airborne endotoxin levels in the indoor environment of five university laboratories in Seoul, South Korea, and examined the micro-environmental factors that influenced endotoxin levels. These included temperature, relative humidity, CO2, CO, illumination, and wind velocity. A total of 174 air samples were collected and analyzed using the kinetic limulus amebocyte lysate assay. Endotoxin levels ranged from <0.001 to 8.90EU/m(3), with an overall geometric mean of 0.240EU/m(3). Endotoxin levels showed significantly negative correlation with temperature (r=-0.529, p<0.001), CO2 (r=-0.213, p<0.001) and illumination (r=-0.538, p<0.001). Endotoxin levels tended to be higher in winter. Endotoxin levels in laboratories with rabbits were significantly higher than those of laboratories with mice. Multivariate regression analysis showed that the environmental factors affecting endotoxin levels were temperature (coefficient=-0.388, p<0.001) and illumination (coefficient=-0.370, p<0.001). Strategies aimed at reducing airborne endotoxin levels in the indoor environments may be most effective if they focus on illumination. PMID:26656510

  4. Identification, expression pattern and potential role of variable lymphocyte receptor Aj-VLRA from Apostichopus japonicus in response to bacterial challenge.

    Science.gov (United States)

    Yang, Lei; Yao, Feng; Ba, Huazhong; Qin, Tong; Luan, Hong; Li, Zhengmin; Hou, Lin; Zou, Xiangyang

    2015-08-01

    The variable lymphocyte receptors (VLRs) are found in jawless vertebrates (agnathans), and specifically recognize bacteria and viruses via their leucine-rich repeats (LRRs). VLRs are believed to be adaptive immune response molecules. Echinoderms do not have adaptive immune systems; however, in the present study, a VLR cDNA named Aj-VLRA was cloned and characterized from sea cucumber, Apostichopus japonicus. The complete cDNA of Aj-VLRA was 3072 bp, including a 1995 bp open reading frame encoding 664 amino acids comprising LRR domains, a predicted transmembrane helix and an N-terminal signal peptide. As determined by quantitative real-time reverse transcription polymerase chain reaction, Aj-VLRA transcripts are ubiquitously expressed in the body wall, longitudinal muscles, intestine and respiratory tree of A. japonicus. The expression level of Aj-VLRA was upregulated after challenge with four common marine bacteria. In situ hybridization showed that the expression of Aj-VLRA was widely distributed in the four tissues, particularly in the cytoplasm of epidermal cells. Recombinantly expressed Aj-VLRA (including the LRR domains) could bind to bacteria including Micrococcus lysodeikticus (Gram+) and Vibrio anguillarum (Gram-). Collectively, the results suggested that Aj-VLRA is related to an innate immune response of A. japonicus. PMID:25896798

  5. Agent-based modeling of endotoxin-induced acute inflammatory response in human blood leukocytes.

    Directory of Open Access Journals (Sweden)

    Xu Dong

    Full Text Available BACKGROUND: Inflammation is a highly complex biological response evoked by many stimuli. A persistent challenge in modeling this dynamic process has been the (nonlinear nature of the response that precludes the single-variable assumption. Systems-based approaches offer a promising possibility for understanding inflammation in its homeostatic context. In order to study the underlying complexity of the acute inflammatory response, an agent-based framework is developed that models the emerging host response as the outcome of orchestrated interactions associated with intricate signaling cascades and intercellular immune system interactions. METHODOLOGY/PRINCIPAL FINDINGS: An agent-based modeling (ABM framework is proposed to study the nonlinear dynamics of acute human inflammation. The model is implemented using NetLogo software. Interacting agents involve either inflammation-specific molecules or cells essential for the propagation of the inflammatory reaction across the system. Spatial orientation of molecule interactions involved in signaling cascades coupled with the cellular heterogeneity are further taken into account. The proposed in silico model is evaluated through its ability to successfully reproduce a self-limited inflammatory response as well as a series of scenarios indicative of the nonlinear dynamics of the response. Such scenarios involve either a persistent (noninfectious response or innate immune tolerance and potentiation effects followed by perturbations in intracellular signaling molecules and cascades. CONCLUSIONS/SIGNIFICANCE: The ABM framework developed in this study provides insight on the stochastic interactions of the mediators involved in the propagation of endotoxin signaling at the cellular response level. The simulation results are in accordance with our prior research effort associated with the development of deterministic human inflammation models that include transcriptional dynamics, signaling, and physiological

  6. Acute endotoxin-induced thymic atrophy is characterized by intrathymic inflammatory and wound healing responses.

    Directory of Open Access Journals (Sweden)

    Matthew J Billard

    Full Text Available BACKGROUND: Productive thymopoiesis is essential for a robust and healthy immune system. Thymus unfortunately is acutely sensitive to stress resulting in involution and decreased T cell production. Thymic involution is a complication of many clinical settings, including infection, malnutrition, starvation, and irradiation or immunosuppressive therapies. Systemic rises in glucocorticoids and inflammatory cytokines are known to contribute to thymic atrophy. Little is known, however, about intrathymic mechanisms that may actively contribute to thymus atrophy or initiate thymic recovery following stress events. METHODOLOGY/PRINCIPAL FINDINGS: Phenotypic, histologic and transcriptome/pathway analysis of murine thymic tissue during the early stages of endotoxemia-induced thymic involution was performed to identify putative mechanisms that drive thymic involution during stress. Thymus atrophy in this murine model was confirmed by down-regulation of genes involved in T cell development, cell activation, and cell cycle progression, correlating with observed phenotypic and histologic thymus involution. Significant gene changes support the hypothesis that multiple key intrathymic pathways are differentially activated during stress-induced thymic involution. These included direct activation of thymus tissue by LPS through TLR signaling, local expression of inflammatory cytokines, inhibition of T cell signaling, and induction of wound healing/tissue remodeling. CONCLUSIONS/SIGNIFICANCE: Taken together, these observations demonstrated that in addition to the classic systemic response, a direct intrathymic response to endotoxin challenge concurrently contributes to thymic involution during endotoxemia. These findings are a substantial advancement over current understanding of thymus response to stress and may lead to the development of novel therapeutic approaches to ameliorate immune deficiency associated with stress events.

  7. Protective Effect of Curcumin on Endotoxin-induced Acute Lung Injury in Rats

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    To investigate the protective effect of curcumin on endotoxin-induced acute lung injury in rats, and explore the underlying mechanisms, 24 male Wistar rats were randomly divided into 4 experimental groups: sham-vehicle (S), sham-curcumin (C), lipopolysaccharide (LPS)-vehicle (L), and curcumin-lipopolysaccharide (C-L) groups. The wet/dry (W/D) weight ratio of the lung and bronchoalveolar lavage (BAL) fluid protein content were used as measures of lung injury. Neutrophil recruitment and activation were evaluated by BAL fluid cellularity and myeloperoxidase (MPO) activity in cell-free BAL and lung tissue. The levels of cytokine-induced neutrophil chemoattractant-1(CINC-1) in lung tissues were measured by ELISA. The histopathological changes of lung tissues were observed by using the HE staining. Our results showed that lung injury parameters, including the wet/dry weight ratio and protein content in BALF, were significantly higher in the L group than in the S group (P<0.01). In the L group, higher numbers of neutrophils and greater MPO activity in cell-free BAL and lung homogenates were observed when compared with the S group (P<0.01).There was a marked increase in CINC-1 levels in lung tissues in response to LPS challenge (P<0.01,L group vs S group). Curcumin pretreatment significantly attenuated LPS-induced changes in these indices. LPS caused extensive morphological lung damage, which was also lessened after curcumin pretreatment. All the above-mentioned parameters in the C group were not significantly different from those of the S group. It is concluded that curcumin pretreatment attenuates LPS-induced lung injury in rats. This beneficial effect of curcumin may involves, in part, inhibition of neutrophilic recruitment and activity, possibly through inhibition of lung CINC-1 expression.

  8. Common studied polymorphisms do not affect plasma cytokine levels upon endotoxin exposure in humans

    DEFF Research Database (Denmark)

    Taudorf, S.; Krabbe, K.S.; Berg, R.M.;

    2008-01-01

    The aim of this study was to investigate to what extent single nucleotide polymorphisms (SNPs) in promoter regions of genes of Toll-like receptor (TLR)-4, tumour necrosis factor (TNF)-alpha, interleukin (IL)-18, interferon (IFN)-gamma, IL-6 and IL-10 affect the cytokine response during a controlled......-607, IFN-gamma+874, IL-6-174, IL-10-592 and IL-10-1082) and endotoxin-induced changes in plasma levels of TNF-alpha, IL-6 and IL-10. IL-18 levels were unaffected by endotoxin. In conclusion, the investigated SNPs did not affect endotoxin-induced low-grade cytokine production of TNF-alpha, IL-6, IL-18 or IL...

  9. Personal Exposure to Particulate Matter and Endotoxin in California Dairy Workers

    Science.gov (United States)

    Garcia, Johnny

    The average number of cows per dairy has increased over the last thirty years, with little known about how this increase may impact occupational exposure. Thirteen California dairies and 226 workers participated in this study throughout the 2008 summer months. Particulate Matter (PM) and endotoxin concentrations were quantified using ambient area based and personal air samplers. Two size fractions were collected, Total Suspended Particulate matter (TSP) and PM 2.5. Differences across dairies were evaluated by placing area based integrated air samplers in established locations on the dairies, e.g. milking parlor, drylot corral, and freestall barns. The workers occupational exposure was quantified using personal air samplers. We analyzed concentrations along with the time workers spent conducting specific job tasks during their shift to identify high exposure job tasks. Biological and chemical analytical methods were employed to ascertain endotoxin concentrations in personal and area based air samples. Recombinant factor C assays (rFC) were used to analyze biologically active endotoxin and gas chromatography coupled with mass spectrometry in tandem (GC-MS/MS) was used to quantify total endotoxin. The PM2.5 concentrations ranged from 2-116 mug/m3 for ambient area concentration and 7-495 mug/m3 for personal concentrations while TSP concentrations ranged from 74-1690 mug/m3 for area ambient concentrations and 191-4950 mug/m3 for personal concentrations. Biologically active endotoxin concentrations in the TSP size fraction from ambient area based samples ranged from 11-2095 EU/m3 and 45-2061 EU/m3 for personal samples. Total endotoxin in the TSP size fraction ranged from 75-10,166 pmol/m3 for area based samples and 34-11,689 pmol/m3 for personal samples. Drylot corrals were found to have higher sample mean concentrations when compared to other locations on the dairies for PM and endotoxin. Re-bedding, of the freestalls, was found to consistently lead to higher personal

  10. Detoxication of endotoxins from the enteric group of bacteria using gamma-irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Tumanian, M.A.; Dunlishcheva, A.P.; Sinilova, N.G.; Ivanov, K.K.

    1975-03-01

    A study was made of the effect of gamma radiation on the toxic properties of endotoxins of the O-somatic antigens and lipopolysaccharides separated from various representatives of the enterobacteriaceae: S. typhi 2 4446, S. paratyphi B42, B. proteus vulgaris 1 and mirabilis 4, E. coli K12 and 5396/38 and Sh. dysenteriae flexneri 516 and 550. An irradiation dose of 1 mrad led to an insignificant decrease in toxicity, while a dose of 3 mrad led to a decrease in toxicity by a factor of 3 to 4 in comparison with the toxicity of the initial preparation. The decrease in toxicity under the effect of irradiation is apparently connected with the lipid part of the endotoxin molecule. The irradiation results in a decrease in the molecular weight of the endotoxin. The infrared spectrum of the molecule shows considerable changes at high radiation doses. (SJR)

  11. The Chemical Composition of Endotoxin Isolated from Intestinal Strain of Desulfovibrio desulfuricans

    Directory of Open Access Journals (Sweden)

    Jolanta Lodowska

    2012-01-01

    Full Text Available Desulfovibrio desulfuricans anaerobes are constituents of human alimentary tract microflora. There are suggestions that they take part in the pathogenesis of periodontitis and some gastrointestinal inflammatory disorders, such as ulcerative colitis or Crohn’s disease. Endotoxin is one of Gram-negative bacteria cellular components that influence these microorganisms pathogenicity. Endotoxin is a lipid-polisaccharide heteropolymer consisting of three elements: lipid A, core oligosaccharide, and O-specific polysaccharide, also called antigen-O. The biological activity of lipopolysaccharide (LPS is determined by its structure. In this study, we show that rhamnose, fucose, mannose, glucose, galactose, heptose, and 2-keto-3-deoxyoctulosonic acid (Kdo are constituents of D. desulfuricans endotoxin oligosaccharide core and O-antigen. Lipid A of these bacteria LPS is composed of glucosamine disaccharide substituted by 3-acyloxyacyl residues: ester-bound 3-(dodecanoyloxytetradecanoic, 3-(hexadecanoyloxytetradecanoic acid, and amide-bound 3-(tetradecanoyloxytetradecanoic acid.

  12. Effect of multiple doses of endotoxin on production of nitric oxide by endothelial cells

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    To study the direct effect of E.Coli endotoxin on the production of nitric oxide by endothelial cells, the second passage of cultured human umbilical cells was stimulated by serial doses of endotoxin (1 g/L, 10 mg/L, 100 μg/L, 10 μg/L, 1 μg/L), and the content of nitric oxide in supematant of culture and the viability of endothelial cells 6 hours after the stimulation were obcerved. The result showed that endotoxin had a slightly inhibitory effect on both the production of nitric oxide and the viability of endothelial cells at low doses (1 μg/L, 10 μg/L, 100 μg/L), especially the dose of 100 μg/L [(608.63±11.64) μmol/L, versus that of unstimulated grouop (629.46±13.36) μmol/L, P<0.05]. While the high doses of endotoxin exerted a big increasing in production of nitric oxide and a big decrease in the viability of endothelial cells, especially the dose of 1 g/L (NO: 722.58 μmol/L±32.18 μmol/L, versus that of unstimulated group P<0.01; viability: 73.63%±8.50%, versus that of unstimulated group, P<0.01). These could be concluded that low doses of endotoxin mainly resulted in functional changes in endothelial cells, such as decrease in relaxing factor (nitrc oxide), while high doses endotoxin exerted lethal effects on endothelial cells accompanied with high production of nitric oxide, which might be related to the death of cells.

  13. Bacterial carbonatogenesis

    International Nuclear Information System (INIS)

    Several series of experiments in the laboratory as well as in natural conditions teach that the production of carbonate particles by heterotrophic bacteria follows different ways. The 'passive' carbonatogenesis is generated by modifications of the medium that lead to the accumulation of carbonate and bicarbonate ions and to the precipitation of solid particles. The 'active' carbonatogenesis is independent of the metabolic pathways. The carbonate particles are produced by ionic exchanges through the cell membrane following still poorly known mechanisms. Carbonatogenesis appears to be the response of heterotrophic bacterial communities to an enrichment of the milieu in organic matter. The active carbonatogenesis seems to start first. It is followed by the passive one which induces the growth of initially produced particles. The yield of heterotrophic bacterial carbonatogenesis and the amounts of solid carbonates production by bacteria are potentially very high as compared to autotrophic or chemical sedimentation from marine, paralic or continental waters. Furthermore, the bacterial processes are environmentally very ubiquitous; they just require organic matter enrichment. Thus, apart from purely evaporite and autotrophic ones, all Ca and/or Mg carbonates must be considered as from heterotrophic bacterial origin. By the way, the carbon of carbonates comes from primary organic matter. Such considerations ask questions about some interpretations from isotopic data on carbonates. Finally, bacterial heterotrophic carbonatogenesis appears as a fundamental phase in the relationships between atmosphere and lithosphere and in the geo-biological evolution of Earth. (author)

  14. Screening and selection of synthetic peptides for a novel and optimized endotoxin detection method.

    Science.gov (United States)

    Mujika, M; Zuzuarregui, A; Sánchez-Gómez, S; Martínez de Tejada, G; Arana, S; Pérez-Lorenzo, E

    2014-09-30

    The current validated endotoxin detection methods, in spite of being highly sensitive, present several drawbacks in terms of reproducibility, handling and cost. Therefore novel approaches are being carried out in the scientific community to overcome these difficulties. Remarkable efforts are focused on the development of endotoxin-specific biosensors. The key feature of these solutions relies on the proper definition of the capture protocol, especially of the bio-receptor or ligand. The aim of the presented work is the screening and selection of a synthetic peptide specifically designed for LPS detection, as well as the optimization of a procedure for its immobilization onto gold substrates for further application to biosensors. PMID:25034430

  15. Interception of the endotoxin-induced arterial hyporeactivity to vasoconstrictors.

    Science.gov (United States)

    Zhang, Shuang; Cui, Ningren; Li, Shanshan; Guo, Lei; Wu, Yang; Zhu, Daling; Jiang, Chun

    2014-07-01

    Septic shock is a severe pathophysiologic condition characterized by vasodilation, hypotension, hypoperfusion, tissue hypoxia, multiple organ failure and death. It is unclear what causes the septic vasodilation that may result from general dysfunction of vascular smooth muscles (VSMs) or selective disruption of vasomotor balances in VSMs. The latter could be due to enhanced vasorelaxation and/or depressed vasoconstriction. Understanding these may lead to pharmacological interventions to septic vasodilation. Therefore, we performed studies in isolated and perfused mesenteric arterial rings. A 20-h exposure of the rings to lipopolysaccharide (LPS, 1 μg/ml) led to hyporeactivity to phenylephrine (PE). However, the responses of the LPS-treated rings to high concentrations of KCl (60 mM) and ATP remained comparable to control rings, suggesting that contractility of VSMs is retained. The hyporeactivity was marginally affected by atropine, indomethacin and L-NAME, suggesting that endothelium-dependent vasorelaxation does not play a major role. In addition to PE, the LPS-treated rings were hyporeactive to dopamine, histamine and angiotensin II. They showed intermediate hyporeactivity to the thromboxane-A2 receptor agonist U46619. Little hyporeactivity to endothelin-1 (ET-1), serotonin (5-HT) and vasopressin was found. ET-1-induced vasoconstriction occurred without endothelium, whereas the effect of 5-HT was endothelium dependent. Although rings were hyporeactive to some of the vasopressors, their vasoconstriction effects were significantly potentiated by PE co-application. Taken together, these data suggest that the endotoxin-induced vasodilation may not result from general dysfunction of VSMs, neither from the endothelium-dependent vasorelaxation. The promising vascular response to various vasoconstrictors found in this study warrants further investigations of therapeutic potentials of these agents. PMID:24792896

  16. Omega-3 fatty acids modulate neonatal cytokine response to endotoxin.

    Science.gov (United States)

    Espiritu, Michael M; Lin, Hong; Foley, Elizabeth; Tsang, Valerie; Rhee, Eunice; Perlman, Jeffrey; Cunningham-Rundles, Susanna

    2016-08-01

    Neonatal immune response is characterized by an uncompensated pro-inflammatory response that can lead to inflammation-related morbidity and increased susceptibility to infection. We investigated the effects of long-chain n-3 polyunsaturated fatty acids (n-3 PUFAs) docosahexaenoic acid (DHA) or eicosapentaenoic acid (EPA) pre-treatment on cytokine secretion to low-concentration endotoxin (lipopolysaccharide, LPS) in THP-1 monocytes and neonatal cord blood (CB) from healthy full-term infants. Pre-treatment of THP-1 cells, with either n-3 PUFA at 25 or 100 μM significantly reduced IL-6, IL-10, and IL-12 secretion while DHA, but not EPA, reduced TNF-α response to LPS. DHA inhibition was stronger compared to EPA and effective at the low concentration. The same concentrations of n-3 PUFAs inhibited IL-12 but not IL-10 cytokine response in whole CB from 9 infants pre-treated for 24 h. To assess clinical relevance for acute response to LPS, the effects of low-concentration DHA at 25 μM or 12.5 μM were assessed before and after LPS exposure of isolated CB mononuclear cells from 20 infants for 1 h. When added before or after LPS, physiologic DHA treatment produced significant concentration-dependent inhibition of TNF-α, IL-6, IL-1β, and IL-8 secretion. The results demonstrate prophylactic and therapeutic modulation of neonatal cytokine response to LPS and provide proof-of-concept that low-concentration administration of n-3 PUFA could attenuate or resolve neonatal inflammatory response. PMID:26812855

  17. Early weaning alters the acute phase immune response to an endotoxin challenge in beef cattle

    Science.gov (United States)

    Previous research indicates that early weaning prior to shipment can reduce transportation-induced increases in acute phase proteins (APP), and can increase subsequent performance in the feedlot. These data suggest that the combination of weaning and transport stress may compromise the immune system...

  18. Early weaning alters the acute phase response to an endotoxin challenge in beef calves

    Science.gov (United States)

    Previous research indicates that early weaning prior to shipment can reduce transportation-induced increases in acute phase proteins (APP), and can increase subsequent performance in the feedlot. These data suggest that the combination of weaning and transport stress may compromise the immune system...

  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

    goats valuable in inflammatory models. We performed a longitudinal study in eight Norwegian dairy goats that received LPS (0.1 μg/kg, Escherichia coli O26:B6) intravenously. A control group of five goats received corresponding volumes of sterile saline. Clinical examinations were performed continuously...... with the acute phase response, type I interferon signaling, LPS cascade and apoptosis, in addition to cytokines and chemokines were targeted. Pro-inflammatory genes, such as IL1B, CCL3 and IL8, were significantly up-regulated. Interestingly, increased mRNA levels of seven interferon stimulated genes (ISGs) were......, 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...

  20. [The role of chronic dental bacterial infections in the aetiopathogenesis of ischaemic heart disease].

    Science.gov (United States)

    Stypułkowska, Jadwiga; Lyszczarz, Robert; Błazowska, Katarzyna

    2002-01-01

    Chronic dental infections, even of low intensity, may cause the development of atherosclerotic changes in arteries, that lead to coronary heart disease. There are many risk factors for atherosclerosis, but the most important are endothelium function disturbances, platelets activation and oxidative changes of plasmatic lipoproteins. Among factors that can induce the epithelium lesions bacterial factor may play an important role. In consequence of the bacterial cell breakdown place the release of endotoxins takes, that lead directly to the damage of endothelial cells. Apart from this direct effect endotoxins activate the fagocytes releasing superoxide reactive radicals, that cause lesions of endothelium. Probably the most widespread chronic bacterial infections in human are the diseases of periodontium and teeth and their inflammatory complications. Oral cavity is colonized by 300-400 bacterial species. In the case of dental bacterial infections bacteriemia occurs after such procedures as tooth extraction, endodontic treatment, therapeutic and hygienic interventions on periodontal tissues. The results of many investigations show the relationship between the oral status (dental and periodontal diseases as chronic oral infections) and disorders of cardiovascular system. PMID:17474623

  1. 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 UNLABELLED: 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

  2. Bacterial Adhesion & Blocking Bacterial Adhesion

    DEFF Research Database (Denmark)

    Vejborg, Rebecca Munk

    2008-01-01

    tract to the microbial flocs in waste water treatment facilities. Microbial biofilms may however also cause a wide range of industrial and medical problems, and have been implicated in a wide range of persistent infectious diseases, including implantassociated microbial infections. Bacterial adhesion...... is the first committing step in biofilm formation, and has therefore been intensely scrutinized. Much however, still remains elusive. Bacterial adhesion is a highly complex process, which is influenced by a variety of factors. In this thesis, a range of physico-chemical, molecular and environmental parameters......, which influence the transition from a planktonic lifestyle to a sessile lifestyle, have been studied. Protein conditioning film formation was found to influence bacterial adhesion and subsequent biofilm formation considerable, and an aqueous extract of fish muscle tissue was shown to significantly...

  3. Bacterial lipases

    NARCIS (Netherlands)

    Jaeger, Karl-Erich; Ransac, Stéphane; Dijkstra, Bauke W.; Colson, Charles; Heuvel, Margreet van; Misset, Onno

    1994-01-01

    Many different bacterial species produce lipases which hydrolyze esters of glycerol with preferably long-chain fatty acids. They act at the interface generated by a hydrophobic lipid substrate in a hydrophilic aqueous medium. A characteristic property of lipases is called interfacial activation, mea

  4. REPRODUCTIVE CONDITION, GLOMERULAR ADENOSINE DIPHOSPHATASE ACTIVITY, AND PLATELET-AGGREGATION IN THE RAT - EFFECT OF ENDOTOXIN

    NARCIS (Netherlands)

    VISSCHER, CA; FAAS, MM; BAKKER, WW; SCHUILING, GA

    1993-01-01

    In experiment A, the activity of the glomerular antithrombotic enzyme adenosine diphosphatase (ADPase) and the sensitivity of this enzyme for endotoxin (1.0 mug/kg BW) in various reproductive conditions of female rats were studied through use of enzyme histochemical methods. In experiment B, the eff

  5. Endotoxin-induced liver damage in rats is minimized by β2- adrenoceptor stimulation

    NARCIS (Netherlands)

    Izeboud, C.A.; Hoebe, K.H.N.; Grootendorst, A.F.; Nijmeijer, S.M.; Miert, A.S. van; Witkamp, R.F.; Rodenburg, R.J.T.

    2004-01-01

    Objective and Design: To investigate the effects of β2- adrenoceptor (β2-AR) stimulation on endotoxin-induced liver damage and systemic cytokine levels in rats. Subjects: Standard male Wistar rats. Treatment: A disease-model of lipolysaccharide (LPS)-induced acute systemic inflammation was used. The

  6. Corticosterone treatment of pregnant low dose endotoxin-treated rats : Inhibition of the inflammatory response

    NARCIS (Netherlands)

    Faas, MM; Slot, K; Koiter, TR; Schuiling, GA

    2000-01-01

    PROBLEM: Can the endotoxin-induced inflammatory response, underlying experimental pre-eclampsia, in pregnant rats be inhibited by corticosterone? METHOD OF STUDY: On day 10 of pregnancy, rats were implanted with pellets containing 25% corticosterone and 75% cholesterol (n = 10) or with 100% choleste

  7. [Interference for Various Quench Agents of Chemical Disinfectants on Detection of Endotoxin Activities in Water].

    Science.gov (United States)

    Zhang, Can; Liu, Wen-jun; Shi, Yun; An, Dai-zhi; Bai, Miao; Xu, Wen

    2015-05-01

    The quenching agents such as histidine, glycine, ascorbic acid, Tween-80, sodium sulfite and sodium hyposulfite are commonly used for quenching the residual disinfectant in water. In this paper, in order to select the optimal type and concentration range of quenching agents prior to the Limulus assays, the interference effects of each quenching agent at different concentrations on endotoxin detection were investigated by the Limulus assays of kinetic-turbidity. Our results identified that, as for 0-1.0% concentration of histidine, ascorbic acid, Tween-80, sodium sulfite (pH unadjusted and pH neutral), interference on the Limulus assays was existed. Hence, these quenching agents could not be applied as neutralizers prior to Limulus assays. Although, there was no interference on endotoxin detection for the glycine, a yellow color, developed by the quenching products of glycine and glutaric dialdehyde, contributed to false positive results. Hence, glycine should not be used as quenching agents in Limulus assays for samples containing glutaric dialdehyde. Compared with other quenching agents as histidine, glycine, ascorbic acid, Tween-80, sodium sulfite, 0-1.0% concentration of sodium hyposulfite elicited no obvious interference, while 1.0%-5.0% concentration of sodium hyposulfite illustrated exhibition effect for endotoxin detection. All in all, compared with other quenching agents as histidine, glycine, ascorbic acid, Tween-80 and sodium sulfite, sodium hyposulfite is suitable for quenching chemicals prior to endotoxin detection and less than 0.5% of concentration is allowable. PMID:26314115

  8. 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...... infusion, it only temporarily attenuated a part of the activated cell-mediated immunity....

  9. Exposure to wood dust and endotoxin in small-scale wood industries in Tanzania.

    NARCIS (Netherlands)

    Rongo, L.M.B.; Msamanga, G.I.; Burstyn, I.; Barten, F.J.M.H.; Dolmans, W.M.V.; Heederik, D.

    2004-01-01

    Workers in small-scale wood industries (SSWI) have increased risks of developing asthma and other respiratory diseases. Wood dust and microbial agents have both been suggested to play a role, but few studies have measured endotoxin exposure in SSWI in Africa. We assessed inhalable dust levels in 281

  10. 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 thuringiensis variety kurstaki encapsulated into killed Pseudomonas fluorescens; exemption from the requirement of a tolerance. 180.1107 Section 180.1107 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) PESTICIDE PROGRAMS TOLERANCES...

  11. Evaluation of a portable test system for assessing endotoxin activity in raw milk.

    Science.gov (United States)

    Suzuki, Yohko; Suzuki, Kazuyuki; Shimamori, Toshio; Tsuchiya, Masakazu; Niehaus, Andrew; Lakritz, Jeffrey

    2016-01-01

    The aim of the present study was to compare endotoxin activities detected in raw milk samples obtained from cattle by a commercially available portable test system (PTS) and traditional microplate limulus amebocyte lysate (LAL)-based assay, which determined activities using a kinetic turbidimetric (KT) assay. Raw milk samples were obtained from 53 and 12 dairy cattle without and with clinical mastitis, respectively. Comparison between the KT and PTS was performed by the Friedman test. The Pearson product moment correlation coefficients were calculated to evaluate associations between any two continuous variables. Linear regression model analysis was also performed to obtain the equation describing the relationship between PTS and KT assay. The endotoxin activities detected in 200- or 400-fold diluted milk samples were similar between PTS and KT assay, whereas a significant difference was observed in 100-fold diluted milk (P<0.001). The results obtained from 200- (r(2)=0.778, P<0.001) and 400-fold diluted milk samples (r(2)=0.945, P<0.001) using PTS correlated with those using KT assay. The median milk endotoxin activities in Gram-positive and Gram-negative clinical mastitis cows were 0.655 and 11,523.5 EU/ml, respectively. The results of the present study suggest that PTS as a simple and easy test to assess endotoxin activity in raw milk is efficient, simple and reproducible. PMID:26279135

  12. Effect of Zingiber officinale and propolis on microorganisms and endotoxins in root canals

    Directory of Open Access Journals (Sweden)

    Lilian Eiko MAEKAWA

    2013-03-01

    Full Text Available The purpose of this study was to evaluate the effectiveness of glycolic propolis (PRO and ginger (GIN extracts, calcium hydroxide (CH, chlorhexidine (CLX gel and their combinations as ICMs (ICMs against Candida albicans, Enterococcus faecalis, Escherichia coli and endotoxins in root canals. Material and Methods After 28 days of contamination with microorganisms, the canals were instrumented and then divided according to the ICM: CH+saline; CLX, CH+CLX, PRO, PRO+CH; GIN; GIN+CH; saline. The antimicrobial activity and quantification of endotoxins by the chromogenic test of Limulus amebocyte lysate were evaluated after contamination and instrumentation at 14 days of ICM application and 7 days after ICM removal. Results and Conclusion After analysis of results and application of the Kruskal-Wallis and Dunn statistical tests at 5% significance level, it was concluded that all ICMs were able to eliminate the microorganisms in the root canals and reduce their amount of endotoxins; however, CH was more effective in neutralizing endotoxins and less effective against C. albicans and E. faecalis, requiring the use of medication combinations to obtain higher success.

  13. Endotoxin and gender modify lung function recovery after occupational organic dust exposure: a 30 year study

    Science.gov (United States)

    Valeri, Linda; Zhang, Feng-ying; Zheng, Bu-Yong; Mehta, Amar J.; Shi, Jing; Su, Li; Brown, Dan; Eisen, Ellen A; Christiani, David C.

    2015-01-01

    OBJECTIVES The purpose of this study is to determine the trajectory of lung function change after exposure cessation to occupational organic dust exposure, and to identify factors that modify improvement. METHODS The Shanghai Textile Worker Study is a longitudinal study of 447 cotton workers exposed to endotoxin-containing dust and 472 silk workers exposed to non-endotoxin-containing dust. Spirometry was performed at 5 year intervals. Air sampling was performed to estimate individual cumulative exposures. The effect of work cessation on FEV1 was modeled using generalized additive mixed effects models to identify the trajectory of FEV1 recovery. Linear mixed effects models incorporating interaction terms were used to identify modifiers of FEV1 recovery. Loss to follow-up was accounted for with inverse probability of censoring weights. RESULTS 74.2% of the original cohort still alive participated in 2011. Generalized additive mixed models identified a non-linear improvement in FEV1 for all workers after exposure cessation, with no plateau noted 25 years after retirement. Linear mixed effects models incorporating interaction terms identified prior endotoxin exposure (p=0.01) and male gender (p=0.002) as risk factors for impaired FEV1 improvement after exposure cessation. After adjusting for gender, smoking delayed the onset of FEV1 gain but did not affect the overall magnitude of change. CONCLUSIONS Lung function improvement after cessation of exposure to organic dust is sustained. Endotoxin exposure and male gender are risk factors for less FEV1 improvement. PMID:25666844

  14. Endotoxin release in cardiac surgery with cardiopulmonary bypass : pathophysiology and possible therapeutic strategies. An update

    NARCIS (Netherlands)

    Kats, Suzanne; Schonberger, Jacques P. A. M.; Brands, Ruud; Seinen, Willem; van Oeveren, Wim

    2011-01-01

    Cardiac surgery with cardiopulmonary bypass provokes a systemic inflammatory response syndrome caused by the surgical trauma itself, blood contact with the non-physiological surfaces of the extracorporeal circuit, endotoxemia, and ischemia. The role of endotoxin in the inflammatory response syndrome

  15. 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

    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...

  16. Endotoxin levels in settled airborne dust in European schools: the HITEA school study.

    NARCIS (Netherlands)

    Jacobs, J.H.; Krop, E.J.M.; Borràs-Santos, A.; Zock, J.P.; Täubel, M.; Hyvärinen, A.; Pekkanen, J.; Doekes, G.; Heederik, D.J.J.

    2014-01-01

    Indoor exposure to microbial agents is known to influence respiratory health. Besides home exposure, exposure in schools can affect respiratory health. In this study, we measured endotoxin in settled dust in primary schools in three European countries from three different geographical regions with d

  17. Immune Challenge Activates Neural Inputs to the Ventrolateral Bed Nucleus of the Stria Terminalis

    OpenAIRE

    Bienkowski, Michael S.; Rinaman, Linda

    2011-01-01

    Hypothalamo-pituitary-adrenal (HPA) axis activation in response to infection is an important mechanism by which the nervous system can suppress inflammation. HPA output is controlled by the hypothalamic paraventricular nucleus (PVN). Previously, we determined that noradrenergic inputs to the PVN contribute to, but do not entirely account for, the ability of bacterial endotoxin (i.e., lipopolysacharide, LPS) to activate the HPA axis. The present study investigated LPS-induced recruitment of ne...

  18. Endotoxin induced chorioamnionitis prevents intestinal development during gestation in fetal sheep.

    Directory of Open Access Journals (Sweden)

    Tim G A M Wolfs

    Full Text Available Chorioamnionitis is the most significant source of prenatal inflammation and preterm delivery. Prematurity and prenatal inflammation are associated with compromised postnatal developmental outcomes, of the intestinal immune defence, gut barrier function and the vascular system. We developed a sheep model to study how the antenatal development of the gut was affected by gestation and/or by endotoxin induced chorioamnionitis.Chorioamnionitis was induced at different gestational ages (GA. Animals were sacrificed at low GA after 2d or 14d exposure to chorioamnionitis. Long term effects of 30d exposure to chorioamnionitis were studied in near term animals after induction of chorioamnionitis. The cellular distribution of tight junction protein ZO-1 was shown to be underdeveloped at low GA whereas endotoxin induced chorioamnionitis prevented the maturation of tight junctions during later gestation. Endotoxin induced chorioamnionitis did not induce an early (2d inflammatory response in the gut in preterm animals. However, 14d after endotoxin administration preterm animals had increased numbers of T-lymphocytes, myeloperoxidase-positive cells and gammadelta T-cells which lasted till 30d after induction of chorioamnionitis in then near term animals. At early GA, low intestinal TLR-4 and MD-2 mRNA levels were detected which were further down regulated during endotoxin-induced chorioamnionitis. Predisposition to organ injury by ischemia was assessed by the vascular function of third-generation mesenteric arteries. Endotoxin-exposed animals of low GA had increased contractile response to the thromboxane A2 mimetic U46619 and reduced endothelium-dependent relaxation in responses to acetylcholine. The administration of a nitric oxide (NO donor completely restored endothelial dysfunction suggesting reduced NO bioavailability which was not due to low expression of endothelial nitric oxide synthase.Our results indicate that the distribution of the tight

  19. Influence of endotoxin-induced sepsis on the requirements of propofol-fentanyl infusion rate in pigs

    DEFF Research Database (Denmark)

    Bollen, Peter; Nielsen, Bjørn J; Toft, Palle

    2007-01-01

    Endotoxin-induced sepsis in pigs is a recognized experimental model for the study of human septic shock. Generally, pigs are brought into general anaesthesia before sepsis is induced. It is our experience that drug dosages of propofol and fentanyl need to be reduced during endotoxin-induced sepsis......, in order to prevent respiratory and cardiovascular depression, but the scientific evidence for this observation is lacking. Therefore, we measured the consumption of propofol and fentanyl at equal level of anaesthesia in pigs with (n = 5) and without (n = 5) endotoxin-induced sepsis, using the cerebral...... state index (CSI) as measure of anaesthetic depth. Infusion rates of propofol (P fentanyl (P fentanyl, whereas...

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

    Directory of Open Access Journals (Sweden)

    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.

  1. Bacterial Ecology

    DEFF Research Database (Denmark)

    Fenchel, Tom

    2011-01-01

    Bacterial ecology is concerned with the interactions between bacteria and their biological and nonbiological environments and with the role of bacteria in biogeochemical element cycling. Many fundamental properties of bacteria are consequences of their small size. Thus, they can efficiently exploit...... biogeochemical processes are carried exclusively by bacteria. * Bacteria play an important role in all types of habitats including some that cannot support eukaryotic life....

  2. [Bacterial vaginosis].

    Science.gov (United States)

    Romero Herrero, Daniel; Andreu Domingo, Antonia

    2016-07-01

    Bacterial vaginosis (BV) is the main cause of vaginal dysbacteriosis in the women during the reproductive age. It is an entity in which many studies have focused for years and which is still open for discussion topics. This is due to the diversity of microorganisms that cause it and therefore, its difficult treatment. Bacterial vaginosis is probably the result of vaginal colonization by complex bacterial communities, many of them non-cultivable and with interdependent metabolism where anaerobic populations most likely play an important role in its pathogenesis. The main symptoms are an increase of vaginal discharge and the unpleasant smell of it. It can lead to serious consequences for women, such as an increased risk of contracting sexually transmitted infections including human immunodeficiency virus and upper genital tract and pregnancy complications. Gram stain is the gold standard for microbiological diagnosis of BV, but can also be diagnosed using the Amsel clinical criteria. It should not be considered a sexually transmitted disease but it is highly related to sex. Recurrence is the main problem of medical treatment. Apart from BV, there are other dysbacteriosis less characterized like aerobic vaginitis of which further studies are coming slowly but are achieving more attention and consensus among specialists. PMID:27474242

  3. Assessing endotoxins in equine-derived snake antivenoms: Comparison of the USP pyrogen test and the Limulus Amoebocyte Lysate assay (LAL).

    Science.gov (United States)

    Solano, Gabriela; Gómez, Aarón; León, Guillermo

    2015-10-01

    Snake antivenoms are parenterally administered; therefore, endotoxin content must be strictly controlled. Following international indications to calculate endotoxin limits, it was determined that antivenom doses between 20 mL and 120 mL should not exceed 17.5 Endotoxin Units per milliliter (EU/mL) and 2.9 EU/mL, respectively. The rabbit pyrogen test (RPT) has been used to evaluate endotoxin contamination in antivenoms, but some laboratories have recently implemented the LAL assay. We compared the capability of both tests to evaluate endotoxin contamination in antivenoms, and we found that both methods can detect all endotoxin concentrations in the range of the antivenom specifications. The acceptance criteria of RPT and LAL must be harmonized by calculating the endotoxin limit as the quotient of the threshold pyrogenic dose and the therapeutic dose and the dose administered to rabbits as the quotient of the threshold pyrogenic dose and the endotoxin limit. Since endotoxins from Gram-negative bacteria exert different pyrogenicity, if contamination occurred, antivenom batches that induce pyrogenic reactions may be found in spite of passing LAL specifications. Although LAL assay can be used to assess endotoxin content throughout the antivenom manufacturing process, we recommend that the release of final products be based on the results of both methods. PMID:26325294

  4. Vascular hyporeactivity to angiotensin II induced by Escherichia coli endotoxin is reversed by Nω-Nitro-L-Arginine, an inhibitor of nitric oxide synthase

    Directory of Open Access Journals (Sweden)

    L. A. RODRIGUES

    2009-01-01

    Full Text Available

    Septic shock or sepsis is reported to be one of the major causes of death when followed by systemic infectious trauma in humans and other mammals. Its development leads to a large drop in blood pressure and a reduction in vascular responsiveness to physiological vasoconstrictors which, if not contained, can lead to death. It is proposed that this vascular response is due to the action of bacterial cell wall products released into the bloodstream by the vascular endothelium and is considered a normal response of the body`s defenses against infection. A reduction in vascular reactivity to epinephrine and norepinephrine is observed under these conditions. In the present study in rats, the aim was to assess whether those effects of hypotension and hyporeactivity are also related to another endogenous vasoconstrictor, angiotensin II (AII. We evaluated the variation in the power of this vasoconstrictor over the mean arterial pressure in anesthetized rats, before and after the establishment of hypotension by Escherichia coli endotoxin (Etx. Our results show that in this model of septic shock, there is a reduction in vascular reactivity to AII and this reduction can be reversed by the inhibitor of nitric oxide synthase, Nω-Nitro-L-Arginine (NωNLA. Our results also suggest that other endogenous factors (not yet fully known are involved in the protection of rats against septic shock, in addition to the L-arginine NO pathway. Keywords: vascular hyporeactivity; NO; rat; angiotensin II; NωNLA Escherichia coli endotoxin.

  5. Endotoxins in the prostatic secretions of chronic prostatitis patients: a need for further biomarkers through the use of proteomics

    Institute of Scientific and Technical Information of China (English)

    Sashi S. Kommu; Stewart Reiss

    2006-01-01

    @@ Dear Sir, Dai et al.[1] must be commended on their useful investigation of the clinical significance of endotoxins in the expressed prostatic secretions (EPS) of chronic prostatitis (CP) patients. However, we take issue with their conclusion.

  6. The role of calcium in endotoxin-induced release of calcitonin gene-related peptide (CGRP) from rat spinal cord

    Institute of Scientific and Technical Information of China (English)

    唐跃明; 韩启德; 王宪

    1997-01-01

    In the present study, the role of calcium in endotoxin-induced CGRP release was studied. 2 .5-50 μg/mL endotoxin and 1 -10 mmol/L caffeine caused concentration-dependent increase of CGRP release from rat spinal cord in vitro. However, no additive effect could he found when caffeine and endotoxin were concomitantly incubated. By using capsaicin, Ca2+-free medium, Omega-Conotoxin, nifedipine, W-7, ryanodine, MgCl2, Tris-ATP, rutheni-um red, the results indicate that the release of CGRP evoked by endotoxin from the sensory fibers of rat spinal cord is dependent on extracellular calcium. After entering into the cell through the N-type calcium channel, calcium binds to calmodulin, and triggers calcium release from intracellular calcium store by activating the caffeine-sensitive but ryan-odine-insensitive mechanism.

  7. Endotoxin as a determinant of asthma and wheeze among rural dwelling children and adolescents: A case–control study

    Directory of Open Access Journals (Sweden)

    Lawson Joshua A

    2012-09-01

    Full Text Available Abstract Background The association between endotoxin exposure and asthma is complex and has been associated with rural living. We examined the relationship between domestic endotoxin and asthma or wheeze among rural school-aged children (6–18 years and assessed the interaction between endotoxin and other characteristics with these outcomes. Methods Between 2005 and 2007 we conducted a case–control study of children 6–18 years in the rural region of Humboldt, Canada. Cases (n = 102 reported doctor-diagnosed asthma or wheeze in the past year. Controls (n = 208 were randomly selected from children without asthma or wheeze. Data were collected to ascertain symptoms, asthma history and indoor environmental exposures (questionnaire, endotoxin (dust collection from the play area floor and child’s mattress, and tobacco smoke exposure (saliva collection. Statistical testing was completed using multiple logistic regression to account for potential confounders and to assess interaction between risk factors. A stratified analysis was also completed to examine the effect of personal history of allergy. Results Among children aged 6–12 years, mattress endotoxin concentration (EU/mg and load (EU/m2 were inversely associated with being a case [odds ratio (OR = 0.44, 95% confidence interval (CI = 0.20-0.98; and OR = 0.38, 95% CI = 0.20-0.75, respectively]. These associations were not observed in older children or with play area endotoxin. Conclusions Our results suggest that endotoxin exposure might be protective for asthma or wheeze. The protective effect is found in younger school-aged, non-allergic children. These results may help explain the inconsistencies in previous studies and suggest that the protective effects of endotoxin in the prevention of atopy and asthma or wheeze are most effective earlier in life.

  8. Changes of gut flora and endotoxin in rats with D-galactosamine-induced acute liver failure

    Institute of Scientific and Technical Information of China (English)

    Lan-Juan Li; Zhong-Wen Wu; Dang-Sheng Xiao; Ji-Fang Sheng

    2004-01-01

    AIM: To investigate the changes of gut microflora and endotoxin levels in rats with acute liver failure (ALF) induced by D-galactosamine (GalN).METHODS: Flora and endotoxin levels in the jejunum, ileum and colon in normal rats (group A) and rats with GalNinduced ALF were determined at 24 h (group B) or 48 h (group C) after GalN injection, as well as the endotoxin level in portal venous blood (PVB) and right ventricle blood (RVB) were determined by chromogenic limulus amoebocyte assay.RESULTS: Intestinal (jejunum, ileum, colon)lactobacillus count was statistically reduced in group B compared with those in group A (3.4±0.3 vs4.9±0.3, 6.1±0.4 vs 8.0±0.3,8.1±0.2 vs 9.3±0.2, P<0.001, P<0.001 and P<0.001respectively) and recovered partially in the group C compared with those in the group B, whereas the count of Enterobacteriaceae in the jejunum, ileum and colon in group B was increased markedly compared with those in the group A (5.1±0.3 vs 3.6±0.2, 6.9±0.5 vs 5.3±0.3,8.7±0.2 vs7.6±0.1, P<0.001, P<0.05 and P<0.05 respectively)and restored partially in the group C compared with those in the group B. The endotoxin level in ileum was increased in the group B compared with those in the group A (111.3±22.8 vs 51.5±8.9, P<0.05). In addition, the endotoxin level in PVB was obviously increased in group B compared with that in the group A (76.8±9.1 vs40.6±7.3,P<0.01) and reduced to the baseline at 48 h (group C).CONCLUSIOM: Severely disturbed gut flora in rats with GalN-induced acute liver failure plays an important role in the elevation of endotoxin level in PVB.

  9. 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

    endotoxin were found to be closely correlated with the plasma concentrations of alanine aminotransferase, gamma-glutamyl transferase, and C-reactive protein in patients with alcoholic liver disease. In conclusion, as IgA located in body tissue was shown to suppress the inflammatory process, enhanced......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......, endotoxin, and acute-phase proteins were measured in patients with different stages of alcoholic liver disease and in healthy controls. Antibodies of type IgA, but not IgG, against fecal endotoxins were significantly increased in patients with alcohol-induced liver disease. IgA antibodies against fecal...

  10. 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.

  11. 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. PMID:25431806

  12. Sexually dimorphic effects of a prenatal immune challenge on social play and vasopressin expression in juvenile rats

    Directory of Open Access Journals (Sweden)

    Taylor Patrick V

    2012-06-01

    Full Text Available Abstract Background Infectious diseases and inflammation during pregnancy increase the offspring’s risk for behavioral disorders. However, how immune stress affects neural circuitry during development is not well known. We tested whether a prenatal immune challenge interferes with the development of social play and with neural circuits implicated in social behavior. Methods Pregnant rats were given intraperitoneal injections of the bacterial endotoxin lipopolysaccharide (LPS – 100 μg /kg or saline on the 15th day of pregnancy. Offspring were tested for social play behaviors between postnatal days 26–40. Brains were harvested on postnatal day 45 and processed for arginine vasopressin (AVP mRNA in situ hybridization. Results In males, LPS treatment reduced the frequency of juvenile play behavior and reduced AVP mRNA expression in the medial amygdala and bed nucleus of the stria terminalis. These effects were not found in females. LPS treatment did not change AVP mRNA expression in the suprachiasmatic nucleus, paraventricular nucleus, or supraoptic nucleus of either sex, nor did it affect the sex difference in the size of the sexually dimorphic nucleus of the preoptic area. Conclusions Given AVP’s central role in regulating social behavior, the sexually dimorphic effects of prenatal LPS treatment on male AVP mRNA expression may contribute to the sexually dimorphic effect of LPS on male social play and may, therefore, increase understanding of factors that contribute to sex differences in social psychopathology.

  13. Alveolar macrophages regulate neutrophil recruitment in endotoxin-induced lung injury

    OpenAIRE

    Beck-Schimmer, B; Schwendener, R.; Pasch, T; Reyes, L.; Booy, C; Schimmer, R C

    2005-01-01

    BACKGROUND: Alveolar macrophages play an important role during the development of acute inflammatory lung injury. In the present study, in vivo alveolar macrophage depletion was performed by intratracheal application of dichloromethylene diphosphonate-liposomes in order to study the role of these effector cells in the early endotoxin-induced lung injury. METHODS: Lipopolysaccharide was applied intratracheally and the inflammatory reaction was assessed 4 hours later. Neutrophil accumulation an...

  14. Alveolar macrophages regulate neutrophil recruitment in endotoxin-induced lung injury

    OpenAIRE

    Reyes Livia; Pasch Thomas; Schwendener Reto; Beck-Schimmer Beatrice; Booy Christa; Schimmer Ralph C

    2005-01-01

    Abstract Background Alveolar macrophages play an important role during the development of acute inflammatory lung injury. In the present study, in vivo alveolar macrophage depletion was performed by intratracheal application of dichloromethylene diphosphonate-liposomes in order to study the role of these effector cells in the early endotoxin-induced lung injury. Methods Lipopolysaccharide was applied intratracheally and the inflammatory reaction was assessed 4 hours later. Neutrophil accumula...

  15. Regulation of TB Vaccine-Induced Airway Luminal T Cells by Respiratory Exposure to Endotoxin

    OpenAIRE

    Chen, Xuerong; Xiu, Fangming; Horvath, Carly N.; Damjanovic, Daniela; Thanthrige-Don, Niroshan; Jeyanathan, Mangalakumari; Xing, Zhou

    2012-01-01

    Tuberculosis (TB) vaccine-induced airway luminal T cells (ALT) have recently been shown to be critical to host defense against pulmonary TB. However, the mechanisms that maintain memory ALT remain poorly understood. In particular, whether respiratory mucosal exposure to environmental agents such as endotoxin may regulate the size of vaccine-induced ALT population is still unclear. Using a murine model of respiratory genetic TB vaccination and respiratory LPS exposure, we have addressed this i...

  16. Effects of the immunomodulator, VGX-1027, in endotoxin-induced uveitis in Lewis rats

    DEFF Research Database (Denmark)

    Mangano, K; Sardesai, N Y; Quattrocchi, C;

    2008-01-01

    VGX-1027 is a novel, low molecular weight, immunomodulatory compound that has shown efficacy against a variety of immuno-inflammatory disease models in animals including autoimmune diabetes in NOD mice, collagen-induced arthritis and chemically induced inflammatory colitis. Here, we have studied ...... the effects of VGX-1027 on the development of endotoxin-induced uveitis (EIU) in male Lewis rats, as a model of inflammatory ocular diseases in humans....

  17. Danaparoid sodium inhibits systemic inflammation and prevents endotoxin-induced acute lung injury in rats

    OpenAIRE

    Hagiwara, Satoshi; Iwasaka, Hideo; Hidaka, Seigo; Hishiyama, Sohei; Noguchi, Takayuki

    2008-01-01

    Introduction Systemic inflammatory mediators, including high mobility group box 1 (HMGB1), play an important role in the development of sepsis. Anticoagulants, such as danaparoid sodium (DA), may be able to inhibit sepsis-induced inflammation, but the mechanism of action is not well understood. We hypothesised that DA would act as an inhibitor of systemic inflammation and prevent endotoxin-induced acute lung injury in a rat model. Methods We used male Wistar rats. Animals in the intervention ...

  18. Structural studies of δ-endotoxin Cry 1 C from Bacillus thuringiensis

    International Nuclear Information System (INIS)

    Full text. The δ-endotoxins are a family of crystal protein by a soil bacterium, Bacillus thuringiensis. The study of these proteins has been of great interest due to their highly specific activity against insects of the orders Lepidoptera, Diptera and Coleoptera. Thus, the δa-endotoxins have been used for more than two decades as biological insecticides to control agricultural pests and, more recently, insects vectors of some diseases. The knowledge of their three-dimensional structures is very important to understand their mechanism of action and their high specificity. To date, the structure of only three proteins of the δ-endotoxins family have been reported: Cry3A, a coleopteran-specific toxin (beetle toxin)1, Cry1Aa, a lepidopteran-specific toxin (butterfly toxin)2 and CytB, a dipteran-specific toxin (mosquito toxin)3 Our work is aimed at the determination of the crystallographic structure by X-ray diffraction of δ-endotoxin Cry1C, also toxic to insects of the Lepidoptera order but towards families other than those affected by Cry1Aa. A comparison between these structures may lead to important conclusions about the reasons for the specificity and would allow the planning of mutants with more efficient activity. The cry1C gene was cloned into an adequate vector and expressed in an acrystalliferous B. thuringiensis strain. After cell culture and sporulation the microcrystals of Cry1C were separated by ultra-centrifugation in sacharose. The protoxin inclusion bodies were activated by commercial trpsin and the protease-resistant core was purified by anion-exchange chromatography. Crystallization experiments are being conducted in order to obtain single crystals suitable for diffraction measurements. We intend to use the Protein Crystallograph Station of the LNLS to collect data as soon as it is available and we have suitable crystals. (author)

  19. Glycyrrhizin attenuates endotoxin- induced acute liver injury after partial hepatectomy in rats

    OpenAIRE

    B. Tang; Qiao, H.; Meng, F.; Sun, X.

    2007-01-01

    Massive hepatectomy associated with infection induces liver dysfunction, or even multiple organ failure and death. Glycyrrhizin has been shown to exhibit anti-oxidant and anti-inflammatory activities. The aim of the present study was to investigate whether glycyrrhizin could attenuate endotoxin-induced acute liver injury after partial hepatectomy. Male Wistar rats (6 to 8 weeks old, weighing 200-250 g) were randomly assigned to three groups of 24 rats each: sham, saline and glycyrrhizin. Rats...

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

    OpenAIRE

    Banun Kusumawardani; Marsetyawan HNE Soesatyo; Djaswadi Dasuki; Widya Asmara

    2012-01-01

    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 gingiv...

  1. Endotoxin, dust and exhaled nitrogen oxide among hand pickers of coffee; a cross-sectional study

    OpenAIRE

    Moen, Bente Elisabeth; Kayumba, Akwilina; Sakwari, Gloria; Mamuya, Simon Henry David; Bråtveit, Magne

    2016-01-01

    Background Primary coffee processing takes place in countries where coffee is grown, and may include hand picking of coffee to remove low quality beans. Hand picking is mostly performed by women. No previous studies on dust and respiratory health have been performed in this occupational group, although studies indicate respiratory problems among other coffee production workers. Findings Our aim was to assess dust and endotoxin exposure among hand pickers in a coffee factory and compare the le...

  2. Effects of the immunomodulator, VGX-1027, in endotoxin-induced uveitis in Lewis rats

    DEFF Research Database (Denmark)

    Mangano, K; Sardesai, N Y; Quattrocchi, C;

    2008-01-01

    VGX-1027 is a novel, low molecular weight, immunomodulatory compound that has shown efficacy against a variety of immuno-inflammatory disease models in animals including autoimmune diabetes in NOD mice, collagen-induced arthritis and chemically induced inflammatory colitis. Here, we have studied...... the effects of VGX-1027 on the development of endotoxin-induced uveitis (EIU) in male Lewis rats, as a model of inflammatory ocular diseases in humans....

  3. Structural studies of {delta}-endotoxin Cry 1 C from Bacillus thuringiensis

    Energy Technology Data Exchange (ETDEWEB)

    Guimaraes, B.G.; Garratt, R.C.; Oliva, G. [Sao Paulo Univ., Sao Carlos, SP (Brazil). Inst. de Fisica; Lemos, M.V.F. [UNESP, Jaboticabal, SP (Brazil). Dept. de Biologia Aplicada Agropecuaria; Arantes, O.M.N. [Universidade Estadual de Londrina, PR (Brazil). Dept. de Biologia Geral

    1996-12-31

    Full text. The {delta}-endotoxins are a family of crystal protein by a soil bacterium, Bacillus thuringiensis. The study of these proteins has been of great interest due to their highly specific activity against insects of the orders Lepidoptera, Diptera and Coleoptera. Thus, the {delta}a-endotoxins have been used for more than two decades as biological insecticides to control agricultural pests and, more recently, insects vectors of some diseases. The knowledge of their three-dimensional structures is very important to understand their mechanism of action and their high specificity. To date, the structure of only three proteins of the {delta}-endotoxins family have been reported: Cry3A, a coleopteran-specific toxin (beetle toxin){sup 1}, Cry1Aa, a lepidopteran-specific toxin (butterfly toxin){sup 2} and CytB, a dipteran-specific toxin (mosquito toxin){sup 3} Our work is aimed at the determination of the crystallographic structure by X-ray diffraction of {delta}-endotoxin Cry1C, also toxic to insects of the Lepidoptera order but towards families other than those affected by Cry1Aa. A comparison between these structures may lead to important conclusions about the reasons for the specificity and would allow the planning of mutants with more efficient activity. The cry1C gene was cloned into an adequate vector and expressed in an acrystalliferous B. thuringiensis strain. After cell culture and sporulation the microcrystals of Cry1C were separated by ultra-centrifugation in sacharose. The protoxin inclusion bodies were activated by commercial trpsin and the protease-resistant core was purified by anion-exchange chromatography. Crystallization experiments are being conducted in order to obtain single crystals suitable for diffraction measurements. We intend to use the Protein Crystallograph Station of the LNLS to collect data as soon as it is available and we have suitable crystals. (author) 3 refs.

  4. Activity of Bacillus thuringiensis D(delta)-endotoxins against codling moth (Cydia pomonella L.) larvae

    NARCIS (Netherlands)

    Boncheva, R.; Dukiandjiev, S.; Minkov, I.; Maagd, de R.A.; Naimov, S.

    2006-01-01

    Solubilized protoxins of nine Cry1 and one hybrid Cry1 ¿-endotoxin from Bacillus thuringiensis were tested for their activity against larvae of the codling moth (Cydia pomonella L). Cry1Da was the most toxic, followed by Cry1Ab, Cry1Ba, and Cry1Ac, while Cry1Aa, Cry1Fa, Cry1Ia, and SN19 were still l

  5. Novel fully-integrated biosensor for endotoxin detection via polymyxin B immobilization onto gold electrodes

    OpenAIRE

    A. Zuzuarregui; S. Arana; E. Pérez-Lorenzo; Sánchez-Gómez, S.; Martínez de Tejada, G.; Mujika, M.

    2013-01-01

    In this paper an electrochemical endotoxin biosensor consisting of an immobilized lipopolysaccharide (LPS) ligand, polymyxin B (PmB), is presented. Several parameters involved both in the device fabrication and in the detection process were analyzed to optimize the ligand immobilization and the interaction between PmB and LPS, aiming at increasing the sensitivity of the sensor. Different electrochemical pre-treatment procedures as well as the functionalization methods were s...

  6. Specificities of monoclonal antibodies against the activated delta-endotoxin of Bacillus thuringiensis var. thuringiensis.

    OpenAIRE

    Huber-Lukac, M; Lüthy, P; Braun, D G

    1983-01-01

    Eight hybrid cell lines secreting monoclonal antibodies directed against the activated delta-endotoxin of Bacillus thuringiensis var. thuringiensis were grown in BALB/c mice. Ascites fluids were collected, and the antibodies were purified by antigen-affinity chromatography. The specificity of each monoclonal antibody for the toxin and protoxin was established by the indirect enzyme-linked immunosorbent assay. All the antibodies consisted of gamma 1 heavy and kappa light chains. They were reac...

  7. Endotoxin contamination of enzyme conjugates used in enzyme-linked immunosorbent assays.

    OpenAIRE

    Bryant, R. E.; Chamovitz, B N; Morse, S A; Apicella, M A; Morthland, V H

    1983-01-01

    The specificity of the enzyme-linked immunosorbent assay(s) is thought to depend on the specificity of the antibody used in the assay system. Therefore, the association of broadly reactive antigens like endotoxin with enzyme conjugates or other enzyme-linked immunosorbent assay reagents has the potential of altering the specificity of reactions in the enzyme-linked immunosorbent assay. Using the Limulus amoebocyte lysate assay, we demonstrated that commercially prepared conjugates of goat ant...

  8. Renal blood flow distribution during E. coli endotoxin shock in dog.

    Science.gov (United States)

    Kirkebø, A; Tyssebotn, I

    1980-04-01

    The effect of endotoxin on renal blood flow distribution was studied in anesthetized dogs. Renal blood flow was measured as hydrogen clearance by platinum electrodes placed in outer and in inner halves of cortex and by electromagnetic flowmeter. Intravenous injection of E. coli endotoxin, 3-5 mg/kg b. wt., promptly reduced arterial blood pressure (AP) and renal blood flow. After a transient increase for 45 min AP and renal blood flow declined to about 50% of the control 2 1/2-3 h after injection. The reduction in outer cortical blood flow (OCF) was not significantly different from the reduction in inner cortical blood flow (ICF). The hematocrit (Hct) increased from 40.1 +/- 3.8% to 54.6 +/- 8%, but mean renal vascular resistance did not change. Total plasma protein concentration was not significantly elevated. A marked local flow variability was observed in some periods during the phase of shock with declining AP and total renal blood flow at high Hct. Thus renal blood flow showed phasic changes, but the OCF/ICF ratio was not changed during endotoxin shock. Local blood flow instability was observed periodically at high Hct. PMID:6998257

  9. Analysis of different coating thickness on new type of planar interdigital sensors for endotoxin detection

    KAUST Repository

    Syaifudin, A. R Mohd

    2013-05-01

    New types of planar interdigital sensors have been fabricated on Silicon/Silicon Dioxide (Si/SiO2) wafers. The sensors were coated with pre-cursor silica functionalized with APTES (3-aminopropyltrietoxysilane) at different thicknesses. All sensors were then immobilized with Polymyxin, B (PmB). PmB is an antimicrobial peptide produced by the Gram-positive bacterium-Bacillus, has been immobilized on the coated sensors because of its specific binding properties to endotoxin. Studies were conducted to analyze the effect of different thicknesses of coatings on the sensitivity and selectivity of the sensors. It was observed sensors coated with 3 layers of coating has better sensitivity and selectivity to the target molecules (endotoxin) compared to sensors with 5 layers of coating. The repeatability and stability of the coated sensors were tested by multiple standard endotoxin measurement and it was observed that the sensors give a good reproducibility and stability up to six continuous measurements before the coating degrades. © 2013 IEEE.

  10. Effect of environmental endotoxin exposure on development of pediatric asthma among Egypytian school children

    Institute of Scientific and Technical Information of China (English)

    Malak Shaheen; Sherin El Sayed; Ahmed Abdel Karim; Alfrid Edward; Magid Ibrahim

    2009-01-01

    Objective: The aim of this study was to assess school indoor exposure to microbial products and prevalence of asthma and allergies in rural and urban children. Methods: This study was carried on a rural and an urban school. Environmental endotoxin level was measured in multiple samples of the ambient indoor air dust collected on special aseptic filter papers from the two schools. For two hundred children history taking, clinical examination, allergen skin prick test and basic pulmonary function test were preformed. Results: Environmental endotoxin levels showed significantly higher mean values (P<0.01) in rural school (3 EU/mg) as compared to the urban school (0.1 EU/mg) with (OR=5.163; 95% CI: 0.95-28). History of allergic symptoms was significantly more in urban than rural students (P=0.01). Mean values of pulmonary function parameters were significantly lower values in urban students compared to rural students. Skin prick test results showed significant reactions to all tested allergens in urban children compared to rural children (P<0.05). Conclusion: There is an inverse association between environmental exposure to endotoxins and susceptibility for allergic manifestations in school children.

  11. Therapeutic Effect of Propofol in the Treatment of Endotoxin-induced Shock in Rats

    Institute of Scientific and Technical Information of China (English)

    高巨; 曾邦雄; 周罗晶; 袁世荧; 张诗海

    2003-01-01

    To assess the potential therapeutic effect of propofol in the treatment of endotoxemia, 76rats were randomly assigned to 5 groups: control group(A), endotoxemic group(B), pre-treatmentgroup(C), simultaneous treatment group(D) and post-treatment group(E). Five h after endotoxininjection, PO2, pH, MAP, plasma concentrations of Nitrite/nitrate (NO2-/NO3-) and mortalityrates were assessed in each group. After the rats were sacrificed, lung tissue was sampled to meas-ure myeloperoxidase (MPO) activity and tumor necrosis factor (TNF)-α contents. It was foundthat endotoxin injection produced progressive hypotension, metabolic acidosis, and a large increasein the plasma NO2-/NO3- concentrations and increased mortality rates in 5h. Endotoxin injectionsignificantly increased MPO activity and TNF-α contents in lung tissue (P< 0. 01 or P < 0. 05 ).These changes response to endotoxin were significantly attenuated in the groups B, C and D. Butthese beneficial effects were blunted in the group E. The results suggest that propofol administra-tion may offer advantages in endotoxemia.

  12. In vivo expression of lipopolysaccharide binding protein and its gene induced by endotoxin

    Institute of Scientific and Technical Information of China (English)

    LI Xu-hong 李旭宏; GONG Jian-ping 龚建平; TU Bing 涂兵; SHI Yu-jun 石毓君; LIU Chang-an 刘长安

    2003-01-01

    Objective: To investigate the expression of lipopolysaccharide binding protein (LBP) and its gene in rats with endotoxemia and explore the role of LBP in the response of host to endotoxin.Methods: Thirty Wistar rats were divided randomly into five groups: the normal group and the endotoxemia groups (1, 3, 6, 12 hours after LPS injection, respectively).The level of plasma endotoxin was determined by the Limulus Amebocyte Lysate assay.The expression of LBP mRNA in hepatic tissue was examined by reverse transcription polymerase chain reaction (RT-PCR).Plasma levels of LBP, tumor necrosis factor (TNF)-α and interleukin (IL)-6 were measured by enzyme-linked immunosorbent assay (ELISA).Morphologic changes of hepatic tissue were observed under transmission electron microscope.Results: The level of plasma endotoxin peaked at 1 h after LPS injection, then declined, but was still higher than that of the normal group at 12 h; intrahepatic expression of LBP mRNA and plasma LBP increased with time after LPS stimulation; TNF-α and IL-6 in plasma increased with upregulation of LBP expression; there were significant differences between the normal group and endotoxemia groups (P<0.05).Activation of Kupffer cells and injury of hepatocytes could be seen in rats with endotoxemia.Conclusions: LPS can upregulate the intrahepatic expression of LBP mRNA and increase the plasma LBP level.Under certain conditions, LBP may enhance the sensitivity of host to the toxic effects of LPS.

  13. Study on bacterial endotexins test for Dipyridamole for injectioon%注射用双嘧达莫细菌内毒素检查法研究

    Institute of Scientific and Technical Information of China (English)

    陈琪

    2012-01-01

    Objective To establish a detecting method for the bacterial endotoxin (BET) in the Dipyridamole for injection. Methods The interference test was conducted with reference to Chinese Pharmacopeia (2010 edition) on BET. Results There was no interfere effect in the detection for bacterial endotoxin in the Dipyridamole for injection which was diluted to 0.36 mg/mL. Conclusion It is reliable and applied for bacterial endotoxin test on pyrogen in Dipyridamole for injection.%目的 建立注射用双嘧达莫细菌内毒素检测法.方法 参照2010年版细菌内毒素检查法进行干扰实验.结果 注射用双嘧达莫稀释至0.36 mg/mL后对细菌内毒素检查无干扰作用.结论 细菌内毒素检查法用于注射用双嘧达莫热源检查可靠适用.

  14. Markers of bacterial translocation in end-stage liver disease

    Institute of Scientific and Technical Information of China (English)

    Ioannis; Koutsounas; Garyfallia; Kaltsa; Spyros; I; Siakavellas; Giorgos; Bamias

    2015-01-01

    Bacterial translocation(BT) refers to the passage of viable bacteria or bacterial products from the intestinal lumen, through the intestinal epithelium, into the systemic circulation and extraintestinal locations. The three principal mechanisms that are thought to be involved in BT include bacterial overgrowth, disruption of the gut mucosal barrier and an impaired host defence.BT is commonly observed in liver cirrhosis and has been shown to play an important role in the pathogenesis of the complications of end stage liver disease, including infections as well as hepatic encephalopathy and hepatorenal syndrome. Due to the importance of BT in the natural history of cirrhosis, there is intense interest for the discovery of biomarkers of BT. To date, several such candidates have been proposed, which include bacterial DNA, soluble CD14, lipopolysaccharides endotoxin, lipopolysaccharide-binding protein, calprotectin and procalcitonin. Studies on the association of these markers with BT have demonstrated not only promising data but, oftentimes, contradictory results. As a consequence, currently, there is no optimal marker that may be used in clinical practice as a surrogate for the presence of BT.

  15. Contribution of complement-stimulated hepatic macrophages and neutrophils to endotoxin-induced liver injury in rats.

    Science.gov (United States)

    Jaeschke, H; Farhood, A; Smith, C W

    1994-04-01

    The role of complement as potential activator for tissue macrophages and neutrophils was investigated in an experimental model of endotoxin-induced liver injury in male Fischer rats. Injection of Salmonella enteritidis endotoxin (1 mg/kg) into Corynebacterium parvum-pretreated animals (7 mg/kg; single dose 6 days before endotoxin) resulted in severe oxidant stress, as indicated by a 37-fold increase of plasma levels of glutathione disulfide (basal concentration, 0.36 +/- 14 mumol/L), accumulation of neutrophils in the liver (600 +/- 31 neutrophils/50 high-power fields) and liver injury (plasma ALT, 1184 +/- 185 U/l; necrosis; 19% +/- 3%) 10 hr after endotoxin. The oxidant stress induced by 1 mg/kg endotoxin in the C. parvum-treated animals was always significantly higher than that in control animals receiving the same dose of endotoxin. Inhibition of complement activation with the soluble complement receptor type 1 attenuated the oxidant stress and liver injury by 50% to 65% but had no effect on hepatic neutrophil accumulation or plasma tumor necrosis factor-alpha levels. Treatment with a monoclonal antibody directed against the alpha-chain of CD11b/CD18 adhesion proteins (clone 17), which was highly effective in attenuating ischemia-reperfusion injury in the liver by reducing the number of neutrophils and functionally inactivating these cells, neither protected against parenchymal cell injury nor affected hepatic neutrophil infiltration in the C. parvum model. We conclude that reactive oxygen derived from complement-stimulated macrophages is critical for the development of liver injury in the C. parvum/endotoxin model. PMID:8138272

  16. Monitoring the effectiveness of root canal procedures on endotoxin levels found in teeth with chronic apical periodontitis

    Directory of Open Access Journals (Sweden)

    Ariane Cassia Salustiano MARINHO

    2014-12-01

    Full Text Available Objective: The aim of this study was to monitor the effectiveness of root canal procedures by using different irrigants and intracanal medication on endotoxin levels found in root canals of teeth with chronic apical periodontitis. Material and Methods: Thirty root canals of teeth with pulpal necrosis associated with periapical lesions were selected and randomly divided into groups according to the irrigants used: GI - 2.5% NaOCl, GII - 2% chlorhexidine (CHX gel, and GIII - saline solution (SS (all, n=10. Samples were collected with sterile/apyrogenic paper points before (S1 and after root canal instrumentation (S2, after use of 17% ethylenediaminetetraacetic acid (EDTA (S3, and after 30 days of intracanal medication (Ca(OH2+SS (S4. A turbidimetric kinetic Limulus Amebocyte Lysate assay was used for endotoxin measurement. Results: Endotoxins were detected in 100% of the root canals investigated (30/30, with a median value of 18.70 EU/mL. After S2, significant median percentage reduction was observed in all groups, irrespective of the irrigant tested: 2.5% NaOCl (99.65% (GI, 2% CHX (94.27% (GII, and SS (96.79% (GIII (all p0.05. Intracanal medication for 30 days was able to significantly reduce residual endotoxins: 2.5% NaOCl (90% (GI, 2% CHX (88.8% (GII, and SS (85.7% (GIII, p<0.05. No differences were found in the endotoxin reduction when comparing s2 and s4 treatment groups. Conclusion: Our findings demonstrated the effectiveness of the mechanical action of the instruments along with the flow and backflow of irrigant enduring root canal instrumentation for the endotoxin removal from root canals of teeth with chronic apical periodontitis. Moreover, the use of intracanal medication for 30 days contributed for an improvement of endotoxin reduction.

  17. Sexually dimorphic effects of neonatal immune system activation with lipopolysaccharide on the behavioural response to a homotypic adult immune challenge.

    Science.gov (United States)

    Tenk, Christine M; Kavaliers, Martin; Ossenkopp, Klaus-Peter

    2008-01-01

    Research has shown that acute immune activation during the early postnatal period with the Gram-negative endotoxin, lipopolysaccharide (LPS), alters a variety of physiological and behavioural processes in the adult animal. For example, neonatal LPS exposure affects disease susceptibility later in life, though these effects appear to be modulated by time of exposure, sex, and immune stimulus. The current study examined sex differences in the effect of neonatal LPS treatment on the locomotor activity response to adult LPS administration. Male and female Long-Evans rats were treated systemically with either LPS (50 microg/kg) or saline (0.9%) on postnatal days 3 and 5. Later in adulthood (postnatal day 92), all animals were subjected to an adult LPS challenge and were injected (i.p.) with 200 microg/kg LPS. Two hours after injection, animals were placed in a non-novel open-field and locomotor activity was assessed for 30 min. Body weights were determined both at the time of injection and 24h later to examine LPS-induced weight loss. Adult males treated neonatally with LPS exhibited significantly less horizontal and vertical activity in response to the LPS challenge relative to males treated neonatally with saline. This effect was not observed in females. Thus, the current study provides important evidence of sexual dimorphism in the long-term effects of neonatal LPS exposure on the responses to an adult homotypic immune challenge in rats. These findings have potential clinical significance given that neonatal exposure to pathogens is a fairly common occurrence and Gram-negative bacteria are a common cause of neonatal bacterial infections.

  18. Nutrition, microbiota, and endotoxin-related diseases in dairy cows Nutrição, microbiota e doenças relacionadas à endotoxina em vacas leiteiras

    Directory of Open Access Journals (Sweden)

    Burim N. Ametaj

    2010-07-01

    Full Text Available In this review article we present an overall summary of the role that high-grain/low forage diets have on rumen composition of microbiota and how changes in the diet affect the release of bacterial cell wall components that are toxic to the host. One of these toxic compounds is lipopolysaccharide or endotoxin, a component of the outer membrane of all Gram-negative bacteria. Moreover, data are provided that support the concept that endotoxin translocates into the blood circulation and show that rumen endotoxin is associated with multiple perturbations of blood variables related to carbohydrate, lipid, and mineral metabolism. In addition, endotoxin induces a general, nonspecific immune response known as acute phase response. We also pinpoint the fact that high-grain diets are associated with distinct clusters of plasma metabolites and immune variables suggesting that changing cereal grain to forage ratio in the diet is very important for the health of dairy cattle. Furthermore, we provide information that support the concept that endotoxin is involved in multiple metabolic diseases such as fatty liver, milk fever, laminitis, retained placenta, displaced abomasum, and downer cow syndrome. More research is warranted to clarify the mechanisms by which nutrition, microbiota, and endotoxin contribute to development of metabolic diseases in dairy cattle. It is concluded that besides the aforementioned causal agents other compounds generated in the gastrointestinal tract such as lipoteichoic acid or methylated amines might be involved in the etiology of several metabolic diseases.Neste artigo de revisão é apresentado um resumo total do papel que a dieta de alto teor de grãos e baixo teor de forragem tem sobre a composição da microbiota do rúmen e como as mudanças na dieta afetam a liberação de componentes da parede celular bacteriana tóxicos ao hospedeiro. Um destes compostos tóxicos é um lipopolisacarídeo ou endotoxina, um componente da

  19. Coordinated Regulation of miR-155 and miR-146a Genes during Induction of Endotoxin Tolerance in Macrophages

    DEFF Research Database (Denmark)

    Doxaki, Christina; Kampranis, Sotirios C; Eliopoulos, Aristides G;

    2015-01-01

    Endotoxin tolerance occurs to protect the organism from hyperactivation of innate immune responses, primarily mediated by macrophages. Regulation of endotoxin tolerance occurs at multiple levels of cell responses and requires significant changes in gene expression. In the process of macrophage ac...

  20. Reproductive condition and the low-dose endotoxin-induced inflammatory response in rats. Glomerular influx of inflammatory cells and expression of adhesion molecules

    NARCIS (Netherlands)

    Faas, MM; Bakker, WW; Valkhof, N; vanderHorst, MCL; Schuiling, GA

    1997-01-01

    These experiments were designed to study the increased sensitivity of pregnant rats to endotoxin. Pregnant (Pr), cyclic (C), and progesterone (P)-treated pseudopregnant rats with or without a decidualized uterus (PSP and DEC rats, respectively) received infusions of an ultra-low dose of endotoxin (1

  1. Endotoxin exposure and lung cancer risk: a systematic review and meta-analysis of the published literature on agriculture and cotton textile workers.

    NARCIS (Netherlands)

    Lenters, V.C.; Basinas, I.; Beane Freeman, L.E.; Boffetta, P.; Checkoway, H.; Coggon, D.; Portengen, L.; Sim, M.; Wouters, I.M.; Heederik, D.; Vermeulen, R.

    2010-01-01

    OBJECTIVE: To examine the association between exposure to endotoxins and lung cancer risk by conducting a systematic review and meta-analysis of epidemiologic studies of workers in the cotton textile and agricultural industries; industries known for high exposure levels of endotoxins. METHODS: Risk

  2. Intracellular mechanisms of hydroquinone toxicity on endotoxin-activated neutrophils.

    Science.gov (United States)

    Hebeda, Cristina Bichels; Pinedo, Fernanda Júdice; Bolonheis, Simone Marques; Ferreira, Zulma F; Muscará, Marcelo Nicolas; Teixeira, Simone Aparecida; Farsky, Sandra Helena Poliselli

    2012-11-01

    Circulating neutrophils promptly react to different substances in the blood and orchestrate the beginning of the innate inflammatory response. We have shown that in vivo exposure to hydroquinone (HQ), the most oxidative compound of cigarette smoke and a toxic benzene metabolite, affects circulating neutrophils, making them unresponsive to a subsequent bacterial infection. In order to understand the action of toxic molecular mechanisms on neutrophil functions, in vitro HQ actions on pro-inflammatory mediator secretions evoked by Escherichia coli lipopolysaccharide (LPS) were investigated. Neutrophils from male Wistar rats were cultured with vehicle or HQ (5 or 10 μM; 2 h) and subsequently incubated with LPS (5 μg/ml; 18 h). Hydroquinone treatment impaired LPS-induced nitric oxide (NO), tumour necrosis factor α (TNF-α), interleukin (IL)-1β and IL-6 secretions by neutrophils. The toxic effect was not dependent on cell death, reduced expression of the LPS receptor or toll-like receptor-4 (TLR-4) or cell priming, as HQ did not induce reactive oxygen species generation or β(2)integrin membrane expression. The action of toxic mechanisms on cytokine secretion was dependent on reduced gene synthesis, which may be due to decreased nuclear factor κB (NF-κB) nuclear translocation. Conversely, this intracellular pathway was not involved in impaired NO production because HQ treatments only affected inducible nitric oxide synthase protein expression and activity, suggesting posttranscriptional and/or posttranslational mechanisms of action. Altogether, our data show that HQ alters the action of different LPS-activated pathways on neutrophils, which may contribute to the impaired triggering of the host innate immune reaction detected during in vivo HQ exposure. PMID:22717997

  3. Endotoxin mediated-iNOS induction causes insulin resistance via ONOO⁻ induced tyrosine nitration of IRS-1 in skeletal muscle.

    Directory of Open Access Journals (Sweden)

    Geneviève Pilon

    Full Text Available BACKGROUND: It is believed that the endotoxin lipopolysaccharide (LPS is implicated in the metabolic perturbations associated with both sepsis and obesity (metabolic endotoxemia. Here we examined the role of inducible nitric oxide synthase (iNOS in skeletal muscle insulin resistance using LPS challenge in rats and mice as in vivo models of endotoxemia. METHODOLOGY/PRINCIPAL FINDINGS: Pharmacological (aminoguanidine and genetic strategies (iNOS⁻/⁻ mice were used to counter iNOS induction in vivo. In vitro studies using peroxynitrite (ONOO⁻ or inhibitors of the iNOS pathway, 1400 W and EGCG were conducted in L6 myocytes to determine the mechanism by which iNOS mediates LPS-dependent insulin resistance. In vivo, both pharmacological and genetic invalidation of iNOS prevented LPS-induced muscle insulin resistance. Inhibition of iNOS also prevented insulin resistance in myocytes exposed to cytokine/LPS while exposure of myocytes to ONOO⁻ fully reproduced the inhibitory effect of cytokine/LPS on both insulin-stimulated glucose uptake and PI3K activity. Importantly, LPS treatment in vivo and iNOS induction and ONOO⁻ treatment in vitro promoted tyrosine nitration of IRS-1 and reduced insulin-dependent tyrosine phosphorylation. CONCLUSIONS/SIGNIFICANCE: Our work demonstrates that iNOS-mediated tyrosine nitration of IRS-1 is a key mechanism of skeletal muscle insulin resistance in endotoxemia, and presents nitrosative modification of insulin signaling proteins as a novel therapeutic target for combating muscle insulin resistance in inflammatory settings.

  4. Pathogen Inactivation of red cells: challenges and opportunities

    Institute of Scientific and Technical Information of China (English)

    Stephen J. Wagner

    2006-01-01

    @@ Introduction Virus inactivation methods for blood have been explored as a means to further reduce the risk from tested agents and to decrease the risk of emerging or variant agents for whom no deferral or effective screening methods are available. Although inactivation methods promise to reduce transfusion-related infectious disease risk, these methods are not perfect. Most techniques for pathogen reduction will not kill bacterial spores, or inactivate bacterial endotoxin, prion protein, or certain non-enveloped viruses whose tightly packed capsid proteins prevent access of the virucidal agent to its nucleic acid target. In addition,various inactivation methods have been known to decrease blood cell yield, affect blood cell recovery or survival, and may pose risk to recipients or blood center workers. My presentation today will review two methods for pathogen inactivation of red cells.

  5. Efficacy of a novel endotoxin adsorber polyvinylidene fluoride fiber immobilized with L-serine ligand on septic pigs

    Institute of Scientific and Technical Information of China (English)

    Jian-ping GAO; Man HUANG; Ning LI; Peng-fei WANG; Huan-lin CHEN; Qiu-ping XU

    2011-01-01

    A novel adsorber, polyvinylidene fluoride matrix immobilized with L-serine ligand (PVDF-Ser), was developed in the present study to evaluate its safety and therapeutic efficacy in septic pigs by extracorporeal hemoperfusion.Endotoxin adsorption efficiency (EAE) of the adsorber was firstly measured in vitro. The biocompatibility and hemodynamic changes during extracorporeal circulation were then evaluated. One half of 16 pigs receiving lipopolysaccharide (Escherichia coli O111 :B4, 5 μg/kg) intravenously in 1 h were consecutively treated by hemoperfusion with the new adsorber for 2 h. The changes of circulating endotoxin and certain cytokines and respiratory function were analyzed.The 72 h-survival rate was assessed eventually. EAE reached 46.3% (100 EU/ml in 80 ml calf serum) after 2 hcirculation. No deleterious effect was observed within the process. The plasma endotoxin, interleukin-6 (IL-6), and tumornecrosis factor-α (TNF-α) levels were decreased during the hemoperfusion. Arterial oxygenation was also improved during and after the process. Furthermore, the survival time was significantly extended (>72 h vs. 47.5 h for median survival time). The novel product PVDF-Ser could adsorb endotoxin with high safety and efficacy. Early use of extracorporeal hemoperfusion with the new adsorber could reduce the levels of circulating endotoxin, IL-6, and TNF-α,besides improve respiratory function and consequent 72 h-survival rate of the septic pigs. Endotoxin removal strategy with blood purification using the new adsorber renders a potential promising future in sepsis therapy.

  6. Enhancement of intrinsic antitumor activity in spore-endotoxin mixtures of Bacillus thuringiensis by exposure to ultraviolet radiation

    International Nuclear Information System (INIS)

    Irradiation of spore-endotoxin mixtures from Bacillus thuringiensis cultures at 254 nm (60 μW cm-2) enhances their intrinsic antitumor potency as well as that of either component. The extent of enhancement depends on the length of exposure (optimum: 35 min) and may thus be due to photochemical changes of the endotoxin protein or/and to photoproduction of additional compounds with antitumor activity. Antitumor effects, expressed as survival rates of C57BL/6 mice inoculated with Lewis' mouse lung carcinoma and subjected to treatments 24 h later, depended on the number of doses of preparations administered (mixture, separated components). (author)

  7. Enhancement of intrinsic antitumor activity in spore-endotoxin mixtures of Bacillus thuringiensis by exposure to ultraviolet radiation

    Energy Technology Data Exchange (ETDEWEB)

    Zamola, B.; Karminski-Zamola, G.; Fuks, Z.; Kubovic, M. (Zagreb Univ. (Yugoslavia)); Wrishcer, M. (Institut Rudjer Boskovic, Zagreb (Yugoslavia))

    1985-03-01

    Irradiation of spore-endotoxin mixtures from Bacillus thuringiensis cultures at 254 nm (60 ..mu..W cm/sup -2/) enhances their intrinsic antitumor potency as well as that of either component. The extent of enhancement depends on the length of exposure (optimum: 35 min) and may thus be due to photochemical changes of the endotoxin protein or/and to photoproduction of additional compounds with antitumor activity. Antitumor effects, expressed as survival rates of C57BL/6 mice inoculated with Lewis' mouse lung carcinoma and subjected to treatments 24 h later, depended on the number of doses of preparations administered (mixture, separated components).

  8. Exercise and IL-6 infusion inhibit endotoxin-induced TNF-alpha production in humans

    DEFF Research Database (Denmark)

    Starkie, Rebecca; Ostrowski, Sisse Rye; Jauffred, Sune;

    2003-01-01

    and atherosclerosis. To test this hypothesis, we performed three experiments in which eight healthy males either rested (CON), rode a bicycle for 3 h (EX), or were infused with recombinant human IL-6 (rhIL-6) for 3 h while they rested. After 2.5 h, the volunteers received a bolus of Escherichia coli...... exercise and rhIL-6 infusion at physiological concentrations inhibit endotoxin-induced TNF-alpha production in humans. Hence, these data provide the first experimental evidence that physical activity mediates antiinflammatory activity and suggest that the mechanism include IL-6, which is produced...

  9. Ligustrazini Inhibits Endotoxin Induced PAI-1 Expression in Human Umbilical Vein Endothelial Cells

    Institute of Scientific and Technical Information of China (English)

    阮秋蓉; 邓仲端; 宋建新

    2001-01-01

    Plasminogen activator inhibitor 1 (PAI-1) is one of important coagulant factors. Endotoxin lipopolysaccharide (LPS) induces thrombosis by stimulating PAI-1 secretion of vascular cells (EC). Using sandwich enzyme-linked immunosorbent assay (ELISA) and Northern blot, was investigated the effects of Chinese medicine ligustrazini on PAI-1 expression in EC and LPS-stimulated EC. The results showed that ligustrazini inhibited both basal and LPS-induced PAI-1 mRNA expression in EC. The effect of ligustrazini on LPS-induced PAI-1 secretion worked in a dose-dependent manner. This study provided theoretic and experimental evidence for use of ligustrazini against septic shock and cardiovascular diseases.

  10. Exposure to airborne microorganisms, dust and endotoxin during processing of peppermint and chamomile herbs on farms.

    Science.gov (United States)

    Skórska, Czesława; Sitkowska, Jolanta; Krysińska-Traczyk, Ewa; Cholewa, Grazyna; Dutkiewicz, Jacek

    2005-01-01

    The aim of this study was to determine the levels of microorganisms, dust and endotoxin in the air during processing of peppermint (Mentha piperita) and chamomile (Matricaria recutita) by herb farmers, and to examine the species composition of airborne microflora. Air samples were collected on glass fibre filters by use of personal samplers on 13 farms owned by herb cultivating farmers, located in Lublin province (eastern Poland). The concentrations of total viable microorganisms (bacteria + fungi) in the farm air during processing of peppermint herb were large, within a range from 895.1-6,015.8 x 10(3) cfu/m(3) (median 1,055.3 x 10(3) cfu/m(3)). During processing of chamomile herb they were much lower and varied within a range from 0.88-295.6 x 10(3) cfu/m(3) (median 27.3 x 10(3) cfu/m(3)). Gram-negative bacteria distinctly prevailed during processing of peppermint leaves, forming 46.4-88.5 % of the total airborne microflora. During processing of chamomile herb, Gram-negative bacteria were dominant at 3 out of 6 sampling sites forming 54.7-75.3 % of total microflora, whereas at the remaining 3 sites the most common were fungi forming 46.2-99.9 % of the total count. The species Pantoea agglomerans (synonyms: Erwinia herbicola, Enterobacter agglomerans ), having strong allergenic and endotoxic properties, distinctly prevailed among Gram-negative isolates. Among fungi, the most common species was Alternaria alternata. The concentrations of airborne dust and endotoxin determined on the examined herb farms were large. The concentrations of airborne dust during peppermint and chamomile processing ranged from 86.7-958.9 mg/m(3), and from 1.1-499.2 mg/m(3), respectively (medians 552.3 mg/m(3) and 12.3 mg/m(3)). The concentrations of airborne endotoxin determined during peppermint and chamomile processing were within a wide range 1.53-208.33 microg/m(3) and 0.005-2604.19 microg/m(3) respectively (medians 57.3 microg/m(3) and 0.96 microg/m(3)). In conclusion, farmers

  11. Application of radio-detoxified endotoxin as adjuvant for experimental foot-and-mouth disease vaccine

    International Nuclear Information System (INIS)

    The immunity enhancing adjuvant activity of radiodetoxified endotoxin (RD-LPS) on the potency of 'C' type foot-and-mouth (FMD) vaccine was tested in different animal species. For radiodetoxification 5 Mrad gamma radiation was used. The suitable quantity of RD-LSP (20 μg per mouse) adjuvated FMD vaccines of lower antigen content better than those of higher one. In cattle and sheep the adjuvant effect of oil + RD + LPS surpassed only slightly that of oil alone. The effect of RD-LPS in pig was very pronounced when applied in small doses but further studies in larger animal populations have to confirm this result. (author)

  12. New Cyt-like δ-endotoxins from Dickeya dadantii: structure and aphicidal activity

    OpenAIRE

    Loth, Karine; Costechareyre, Denis; Effantin, Géraldine; Rahbé, Yvan; Condemine, Guy; Landon, Céline; DA SILVA, Pedro

    2015-01-01

    International audience; In the track of new biopesticides, four genes namely cytA, cytB, cytC and cytD encoding proteins homologous to Bacillus thuringiensis (Bt) Cyt toxins have been identified in the plant pathogenic bacteria Dickeya dadantii genome. Here we show that three Cyt-like d-endotoxins from D. dadantii (CytA, CytB and CytC) are toxic to the pathogen of the pea aphid Acyrthosiphon pisum in terms of both mortality and growth rate. The phylogenetic analysis of the comprehensive set o...

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

    Directory of Open Access Journals (Sweden)

    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.

  14. Inflammatory responses in epithelia: endotoxin-induced IL-6 secretion and iNOS/NO production are differentially regulated in mouse mammary epithelial cells

    Directory of Open Access Journals (Sweden)

    Talhouk Rabih S

    2010-11-01

    Full Text Available Abstract Background IL-6 is a pro-inflammatory cytokine that signals via binding to a soluble or membrane bound receptor, while nitric oxide (NO, an oxidative stress molecule, diffuses through the cell membrane without a receptor. Both mediators signal through different mechanisms, yet they are dependent on NFκB. We proposed that both mediators are co-induced and co-regulated in inflamed mammary epithelial cells. Methods SCp2 mammary epithelial cells were treated with bacterial endotoxin (ET for different time periods and analyzed for induction of IL-6 secretion and NO production by ELISA and Griess reaction, respectively. The expression of IL-6 and induced NO synthase (iNOS was assayed by real time PCR and/or western immunoblots, and the activation of NFκB was assayed by immunobinding assay. To investigate the role of mammary cell microenvironment (cell-substratum or interaction of mammary epithelial cell types; critical to mammary development, function, and disease in modulation of the inflammatory response, SCp2 cells were cultured with or without extracellular matrix (EHS or in coculture with their myoepithelial counterpart (SCg6, and assayed for ET-induced IL-6 and NO. Results Endotoxin induced NFκB activation at 1 h after ET application. IL-6 secretion and NO production were induced, but with unexpected delay in expression of mRNA for iNOS compared to IL-6. NFκB/p65 activation was transient but NFκB/p50 activation persisted longer. Selective inhibition of NFκB activation by Wedelolactone reduced ET-induced expression of IL-6 mRNA and protein but not iNOS mRNA or NO production, suggesting differences in IL-6 and iNOS regulation via NFκB. SCp2 cells in coculture with SCg6 but not in presence of EHS dramatically induced IL-6 secretion even in the absence of ET. ET-induced NO production was blunted in SCp2/SCg6 cocultures compared to that in SCp2 alone. Conclusions The differential regulation of IL-6 and iNOS together with the

  15. Gustatory-mediated avoidance of bacterial lipopolysaccharides via TRPA1 activation in Drosophila

    Science.gov (United States)

    Soldano, Alessia; Alpizar, Yeranddy A; Boonen, Brett; Franco, Luis; López-Requena, Alejandro; Liu, Guangda; Mora, Natalia; Yaksi, Emre; Voets, Thomas; Vennekens, Rudi; Hassan, Bassem A; Talavera, Karel

    2016-01-01

    Detecting pathogens and mounting immune responses upon infection is crucial for animal health. However, these responses come at a high metabolic price (McKean and Lazzaro, 2011, Kominsky et al., 2010), and avoiding pathogens before infection may be advantageous. The bacterial endotoxins lipopolysaccharides (LPS) are important immune system infection cues (Abbas et al., 2014), but it remains unknown whether animals possess sensory mechanisms to detect them prior to infection. Here we show that Drosophila melanogaster display strong aversive responses to LPS and that gustatory neurons expressing Gr66a bitter receptors mediate avoidance of LPS in feeding and egg laying assays. We found the expression of the chemosensory cation channel dTRPA1 in these cells to be necessary and sufficient for LPS avoidance. Furthermore, LPS stimulates Drosophila neurons in a TRPA1-dependent manner and activates exogenous dTRPA1 channels in human cells. Our findings demonstrate that flies detect bacterial endotoxins via a gustatory pathway through TRPA1 activation as conserved molecular mechanism. DOI: http://dx.doi.org/10.7554/eLife.13133.001 PMID:27296646

  16. Alveolar macrophages regulate neutrophil recruitment in endotoxin-induced lung injury

    Directory of Open Access Journals (Sweden)

    Reyes Livia

    2005-06-01

    Full Text Available Abstract Background Alveolar macrophages play an important role during the development of acute inflammatory lung injury. In the present study, in vivo alveolar macrophage depletion was performed by intratracheal application of dichloromethylene diphosphonate-liposomes in order to study the role of these effector cells in the early endotoxin-induced lung injury. Methods Lipopolysaccharide was applied intratracheally and the inflammatory reaction was assessed 4 hours later. Neutrophil accumulation and expression of inflammatory mediators were determined. To further analyze in vivo observations, in vitro experiments with alveolar epithelial cells and alveolar macrophages were performed. Results A 320% increase of polymorphonuclear leukocytes in bronchoalveolar lavage fluid was observed in macrophage-depleted compared to macrophage-competent lipopolysaccharide-animals. This neutrophil recruitment was also confirmed in the interstitial space. Monocyte chemoattractant protein-1 concentration in bronchoalveolar lavage fluid was significantly increased in the absence of alveolar macrophages. This phenomenon was underlined by in vitro experiments with alveolar epithelial cells and alveolar macrophages. Neutralizing monocyte chemoattractant protein-1 in the airways diminished neutrophil accumulation. Conclusion These data suggest that alveolar macorphages play an important role in early endotoxin-induced lung injury. They prevent neutrophil influx by controlling monocyte chemoattractant protein-1 production through alveolar epithelial cells. Alveolar macrophages might therefore possess robust anti-inflammatory effects.

  17. Effect of UHMWPE particle size, dose, and endotoxin on in vitro macrophage response.

    Science.gov (United States)

    Alley, Carie; Haggard, Warren; Smith, Richard

    2014-01-01

    Ultra-high molecular weight polyethylene wear debris generated by a prosthetic hip or knee has been linked to osteolysis and the limited lifespan of the implant. However, research results are conflicting with regard to which characteristics of the polyethylene wear debris are most inflammatory. The goal of this study was to determine whether particle size, number, and the presence of endotoxin significantly contribute to increased secretion of pro-inflammatory mediators by macrophages in vitro in response to polyethylene wear debris generated by a hip simulator. The results show that the prevailing inflammatory factor is endotoxin. The macrophages released only minimal levels of TNF-α and IL-6 in response to cleaned polyethylene particles, but these cytokines were released in significantly higher amounts in response to particles spiked with lipopolysaccharide (LPS). The number (up to 500 particles per cell) and size of the particles tested in this study had no significant influence on any of the measured outputs (macrophage viability, TNF-α, IL-6, or PGE₂) unless associated with LPS.

  18. Adrenergic support during anesthesia in experimental endotoxin shock: norepinephrine versus dobutamine.

    Science.gov (United States)

    Van der Linden, P; Gilbart, E; Engelman, E; de Rood, M; Vincent, J L

    1991-02-01

    The effects of norepinephrine and dobutamine were compared during endotoxin shock in dogs anesthetized either with enflurane (E: 1.5%, N = 12) or with i.v. ketamine (K: 5 mg.kg-1 + 0.2 mg.kg-1.min-1, N = 12). An i.v. bolus of 1.5 mg.kg-1 E. coli endotoxin was followed by saline infusion to restore left-sided filling pressures at baseline. With E, heart rate, mean arterial pressure and stroke index decreased (P less than 0.01). The decrease in oxygen delivery (DO2) and in oxygen consumption (VO2) was associated with an increase in blood lactate. In contrast, K anesthesia was associated with remarkable hemodynamic stability. DO2 was well maintained, VO2 decreased (P less than 0.01) and blood lactate did not change. Under E anesthesia, mean arterial pressure increased more with norepinephrine and heart rate increased more with dobutamine. Under K anesthesia, cardiac index, stroke index and left ventricular stroke work index increased similarly with both agents. In both groups DO2 and VO2 increased markedly. The amount of fluid infused was higher with dobutamine than with norepinephrine. Thus, enflurane but not ketamine had depressant cardiovascular effects at the doses used in this model. With both anesthetics, norepinephrine and dobutamine could effectively improve cardiac function. Dobutamine could therefore represent a valuable alternative to norepinephrine for cardiovascular support during anesthesia in septic shock. PMID:2024562

  19. Targeting naproxen to non-parenchymal liver cells protects against endotoxin induced liver damage.

    Science.gov (United States)

    Lebbe, C; Reichen, J; Wartna, E; Sägesser, H; Poelstra, K; Meijer, D K

    1997-01-01

    Non-steroidal anti-inflammatory drugs (NSAID's) could be of value in the treatment of liver disease; however, their use in this situation is limited by renal side effects. Therefore, we explored whether naproxen covalently bound to human serum albumin NAP-HSA) was able to reduce toxicity in an acute model of liver disease induced by endotoxin in rats pretreated with Corynebacterium parvum. In the isolated perfused liver of such animals endotoxin induced cholestasis (0.62 +/- 0.05 vs. 0.24 +/- 0.09 microliter.min-1.g liver-1; p < 0.05), increased vascular resistance (11300 +/- 400 vs. 311000 +/- 2000 dyn.s.cm-5; p < 0.05) and alanine aminotransferase release (22 +/- 9 vs. 149 +/- IU/l; p < 0.05). At the highest dose tested (22 mg/kg, corresponding to 6.0 mumoles naproxen), NAP-HSA normalized ALT release (21 +/- 10 IU/l: p < 0.05) while an equimolar amount of non-targeted naproxen was only partially effective (56 +/- 19 IU/l). A conventional dose of naproxen similarly prevented transaminase release. Cholestasis and increased vascular resistance were also prevented by NAP-HSA. Drug targeting by linking drugs to proteins is a potentially useful approach to maximizing drug effect while minimizing adverse events; this could be particularly useful for compounds with potentially serious adverse effects in patients with chronic liver disease such as the nonsteroidal anti-inflammatory agents used in the present study. PMID:9169987

  20. Large-scale production of endotoxin-free plasmids for transient expression in mammalian cell culture.

    Science.gov (United States)

    Rozkov, Aleksei; Larsson, Bert; Gillström, Stefan; Björnestedt, Robert; Schmidt, Stefan R

    2008-02-15

    Transient expression of recombinant proteins in mammalian cell culture in a 100-L scale requires a large quantity of plasmid that is very labour intensive to achieve with shake flask cultures and commercially available plasmid purification kits. In this paper we describe a process for plasmid production in 100-mg scale. The fermentation is carried out in a 4-L fed-batch culture with a minimal medium. The detection of the end of batch and triggering the exponential (0.1 h(-1)) feed profile was unattended and controlled by Multi-fermenter Control System. A restricted specific growth rate in fed-batch culture increased the specific plasmid yield compared to batch cultures with minimal and rich media. This together with high biomass concentration (68-107 g L(-1) wet weight) achieves high volumetric yields of plasmid (95-277 mg L(-1) depending on the construct). The purification process consisted of alkaline lysis, lysate clarification and ultrafiltration, two-phase extraction with Triton X-114 for endotoxin removal, anion-exchange chromatography as a polishing step, ultrafiltration and sterile filtration. Both fermentation and purification processes were used without optimisation for production of four plasmids yielding from 39 to 163 mg of plasmids with endotoxin content of 2.5 EU mg(-1) or less. PMID:17680665

  1. Endotoxin down-modulates granulocyte colony-stimulating factor receptor (CD114) on human neutrophils.

    Science.gov (United States)

    Hollenstein, U; Homoncik, M; Stohlawetz, P J; Marsik, C; Sieder, A; Eichler, H G; Jilma, B

    2000-07-01

    During infection, the development of nonresponsiveness to granulocyte colony-stimulating factor (G-CSF) may be influenced by the down-modulation of G-CSF receptor (G-CSFR) by cytokines. This down-modulation was studied during experimental human endotoxemia. Healthy volunteers received either 2 ng/kg endotoxin (lipopolysaccharide [LPS], n=20) or placebo (n=10) in a randomized, controlled trial. Endotoxin infusion increased the mean fluorescence intensity of the neutrophil activation marker CD11b >300% after 1 h (P<.001 vs. placebo). LPS infusion down-modulated G-CSFR expression in as early as 60 min (-17%; P=.001 vs. placebo). Down-modulation was almost maximal at 90 min and persisted for 6 h (-50% from baseline; P<.0001 vs. placebo). Plasma levels of G-CSF started to increase only after G-CSFR down-modulation had occurred and peaked 37-fold above baseline at 4 h (P<.0001 vs. placebo). In conclusion, LPS down-modulates G-CSFR expression in humans, which may render neutrophils less responsive to the effects of G-CSF and, thereby, compromise host defense mechanisms.

  2. 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 effect of endotoxin on the controllability of cardiac rhythm in rats

    International Nuclear Information System (INIS)

    Decreased heart rate variability (HRV) has both diagnostic and prognostic value in patients with sepsis. However, it is not known whether reduced HRV in sepsis reflects an altered input from the autonomic nervous system or a remodeling of the cardiac pacemaker cells by inflammatory mediators. The present study aimed to investigate the effect of endotoxin on the heart rate dynamics of a denervated isolated heart in rats. Saline or endotoxin was injected into rats and their hearts were isolated and perfused. Atrial electrical activity was recorded and memory length in the time-series was assessed using inverse statistical analysis. Memory was defined as a statistical feature that lasts for a period of time and distinguishes the time-series from a random process. Endotoxaemic hearts exhibited a prolonged memory compared to the controls with respect to observing rare events. This indicates that a sudden decelerating event could potentially affect the cardiac rhythm of an endotoxaemic heart for a longer time than the controls. The prolongation of memory is indirectly linked to a reduced controllability in a complex system; therefore our data may provide evidence for a reduced controllability in cardiac rhythm following endotoxaemia. (paper)

  3. Evaluation of the antipyretic effect of ketorolac, acetaminophen, and placebo in endotoxin-induced fever.

    Science.gov (United States)

    Vargas, R; Maneatis, T; Bynum, L; Peterson, C; McMahon, F G

    1994-08-01

    The authors studied the antipyretic effect of three intramuscular doses of ketorolac (15, 30, and 60 mg), acetaminophen 650 mg PO, and placebo in healthy male volunteers using an endotoxin-induced fever model. In this double-blind, double-dummy, parallel study, subjects were assigned randomly with equal probability to one of the above treatment groups. Thirty minutes after study medication administration, a 20 unit per kilogram dose of reference standard endotoxin (RSE) was administered intravenously, and temperature was determined every 15 minutes for an 8-hour period. Compared with placebo, all active treatment groups demonstrated a statistically significant reduction in both adjusted area under the temperature-by-time curve (AAUC) and the maximum increase over baseline temperature (dTmax). Furthermore, the 30 mg intramuscular dose of ketorolac demonstrated approximately the same antipyretic activity as the 650 mg oral dose of acetaminophen, and there was a statistically significant dose response across the three ketorolac doses studied (P < .0001). The majority of side effects reported during this study were symptoms associated with fever, including chills, headache, myalgia, and dizziness, all of which are effects of RSE. The frequency of side effects tended to be less in the treatment groups with the greatest antipyretic activity. PMID:7962674

  4. The Level of Vasopressin is not solely resulted from the Concentration of Endotoxin but Proportional to Creatinine in Dogs with Pyometra

    Directory of Open Access Journals (Sweden)

    W. Y. Shia1, K. C. Tung1, 2, S. C. Chang1, 2, C. H. Yang1, 2, C. H. Lee2, C. C. Chou1 and W. M. Lee1, 2*

    2011-04-01

    Full Text Available Pyometra is one of the most common reproductive disorder characterized as fluid accumulation mainly pus and Gram-negative bacteria isolated from uterus in bitches. Impaired function of vasopressin is found in the disease. The impact of the circulating endotoxin concentration on vasopressin involved water regulation is still unclear. To document the effect of endotoxin on vasopressin-involved water regulation in dogs with pyometra, blood samples were collected for examination of circulating endotoxin concentration, osmolarity values, vasopressin concentrations, blood urea nitrogen (BUN, and creatinine. The results indicated that the concentration change of endotoxin was contrary to BUN and creatinine in dogs with pyometra. The trends of concentrations change of BUN and creatinine were similar with osmolarity and vasopressin. Pyometric dogs with high level of circulating endotoxin had significantly lower (P 20 mg/dL and creatinine (> 1.5 mg/dL also had plasma vasopressin significantly higher (P<0.05 than those with low levels of BUN and creatinine and control dogs. The overall results suggested that elevated vasopressin was not mainly associated with the circulating endotoxin concentration and some other factors may collaborate with endotoxin causing impaired function of vasopressin.

  5. OK-432 reduces mortality and bacterial translocation in irradiated and granulocyte-colony stimulating factor (G-CSF)-treated mice

    Energy Technology Data Exchange (ETDEWEB)

    Nose, Masako; Uzawa, Akiko; Ogyu, Toshiaki [National Inst. of Radiological Sciences, Chiba (Japan); Suzuki, Gen

    2001-06-01

    Acute radiation induces bacterial translocation from the gut, followed by systemic infection and sepsis. In order to reduce the mortality after acute whole body irradiation, it is essential to control bacterial translocation. In this study, we established a bacterial translocation assay as a sensitive method to detect minor mucosal injury by radiation. By utilizing this assay, we evaluated the adverse effects, if any, of hematopoietic reagents on the mucosal integrity in the respiratory and gastro-intestinal tracts. Bacterial translocation to the liver and spleen occurred after whole-body irradiation if the dose exceeded 6 Gy. The administration of G-CSF unexpectedly increased the bacterial translocation in 8 Gy-irradiated mice. The pharmaceutical preparation of low-virulent Streptococcus pyogenes, OK-432, significantly reduced the endotoxin levels in peripheral blood without any reduction of bacterial translocation. A combined treatment with G-CSF and OK-432 decreased bacterial translocation and prevented death. This result indicates that the early administration of G-CSF has an adverse effect on bacterial translocation, and that a combined treatment of G-CSF and OK-432 attenuates the adverse effect of G-CSF and improves the survival rate after acute irradiation. (author)

  6. Prevention of bacterial adhesion

    DEFF Research Database (Denmark)

    Klemm, Per; Vejborg, Rebecca Munk; Hancock, Viktoria

    2010-01-01

    Management of bacterial infections is becoming increasingly difficult due to the emergence and increasing prevalence of bacterial pathogens that are resistant to available antibiotics. Conventional antibiotics generally kill bacteria by interfering with vital cellular functions, an approach that ...

  7. Endotoxin adsorption therapy using polymyxin B-immobilized fiber as a treatment for septic shock-associated severe acute cholangitis.

    Science.gov (United States)

    Inoue, Yoshihiro; Fujino, Yasuhisa; Onodera, Makoto; Kikuchi, Satoshi; Takahashi, Gaku; Kojika, Masahiro; Endo, Shigeatsu

    2013-10-01

    The application of endotoxin adsorption therapy for severe acute cholangitis is controversial. We present a survival case of septic shock and multiple organ failure due to severe acute cholangitis. The patient was treated by endotoxin adsorption therapy using polymyxin B-immobilized fiber because he continued to remain in shock even after successful endoscopic nasobiliary drainage. The patient was an 84-year-old male diagnosed with acute cholangitis and acute pancreatitis who was transferred to our department because of shock and severe dyspnea. The patient had already developed acute respiratory failure, acute renal failure, and disseminated intravascular coagulation. We performed endoscopic nasobiliary drainage immediately, but the patient continued to remain in shock and plasma endotoxin level was markedly elevated at 133.6 pg/mL. Therefore, we performed direct hemoperfusion with polymyxin B-immobilized fiber. On starting the hemoperfusion, blood pressure and urine volume increased, and the plasma endotoxin level reduced considerably. On the basis of our experience in this case, we think that direct hemoperfusion with polymyxin B-immobilized fiber may be a useful modality in the management of severe acute cholangitis.

  8. Different domains of Bacillus thuringiensis delta-endotoxins can bind to insect midgut membrane proteins on ligand blots

    NARCIS (Netherlands)

    Maagd, de R.A.; Klei, van der H.; Bakker, P.L.; Stiekema, W.J.; Bosch, D.

    1996-01-01

    We investigated the role of the constituent domains of the CryIA(b) and CryIA(c) δ-endotoxins in binding to midgut epithelial cell membrane proteins of Spodoptera exigua and Manduca sexta on ligand blots. A collection of wild- type and CryIC-CryIA hybrid toxins was used for this purpose. As demonstr

  9. Enteral glutamine pretreatment does not decrease plasma endotoxin level induced by ischemia-reperfusion injury in rats

    Institute of Scientific and Technical Information of China (English)

    Arda Demirkan; Erkin Orazakunov; Berna Savas; M Ayhan Kuzu; Mehmet Melli

    2008-01-01

    AIM: To investigate whether oral glutamine pretreatment prevents impairment of intestinal mucosal integrity during ischemia-reperfusion (I/R) in rats. METHODS: The study was performed as two series with 40 rats in each. Each series of animals was divided into four groups. The first group was used as a control. Animals in the second group were only pretreated with oral glutamine, 1 g/kg for 4 d. The third group received a normal diet, and underwent intestinal I/R, while the fourth group was pretreated with oral glutamine in the same way, and underwent intestinal I/R. Intestinal mucosal permeability to 51Cr-labeled EDTA was measured in urine in the first series of animals. In the second series, histopathological changes in intestinal tissue and plasma endotoxin levels were evaluated. RESULTS: Intestinal I/R produced a significant increase in intestinal permeability, plasma endotoxin level and worsened histopathological alterations. After intestinal I/R, permeability was significantly lower in glutamine- treated rats compared to those which received a normal diet. However, no significant change was observed in plasma endotoxin levels or histopathological findings. CONCLUSION: Although glutamine pretreatment seems to be protective of intestinal integrity, upon I/R injury, such an effect was not observable in the histopathological changes or plasma endotoxin level.

  10. A hybrid Bacillus thuringiensis delta-endotoxin gene gives resistance against a coleopteran and a lepidopteran pest in transgenic potato

    NARCIS (Netherlands)

    Naimov, S.; Dukiandjiev, S.; Maagd, de R.A.

    2003-01-01

    Expression of Bacillus thuringiensis delta-endotoxins has proven to be a successful strategy for obtaining insect resistance in transgenic plants. Drawbacks of expression of a single resistance gene are the limited target spectrum and the potential for rapid adaptation of the pest. Hybrid toxins wit

  11. The immune response during the luteal phase of the ovarian cycle : increasing sensitivity of human monocytes to endotoxin

    NARCIS (Netherlands)

    Bouman, Annechien; Moes, H; Heineman, MJ; de Leij, LFMH; Faas, MM

    2001-01-01

    Objective: To test the hypothesis that during the luteal phase of the human ovarian cycle, as compared with the follicular phase, the percentage of cytokines producing peripheral monocytes after in vitro stimulation with endotoxin is increased. Design: Prospective study. Setting: Academic research i

  12. An interleukin-1 receptor antagonist blocks lipopolysaccharide-induced colony-stimulating factor production and early endotoxin tolerance.

    OpenAIRE

    Henricson, B E; Neta, R; Vogel, S N

    1991-01-01

    In this report, administration of a recombinant interleukin-1 receptor antagonist protein to mice was found to inhibit induction of colony-stimulating factor as well as induction of early endotoxin tolerance by lipopolysaccharide. These findings provide direct evidence that interleukin-1 is an intermediate in these two lipopolysaccharide-induced phenomena.

  13. 40 CFR 180.1108 - Delta endotoxin of Bacillus thuringiensis variety San Diego 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 thuringiensis variety San Diego encapsulated into killed Pseudomonas fluorescens; exemption from the requirement of a tolerance. 180.1108 Section 180.1108 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) PESTICIDE PROGRAMS TOLERANCES...

  14. Determinants of house dust, endotoxin, and β-(1→3)-d-glucan in homes of Danish children

    DEFF Research Database (Denmark)

    Holst, Gitte Juel; Høst, Arne; Doekes, G;

    2015-01-01

    ) assessed their determinants; (iv) Finally, we compared our findings with previous European studies. Bedroom floor dust was analyzed for endotoxin content by the kinetic limulus amoebocyte lysate assay and for β-(1→3)-d-glucan by the inhibition enzyme immunoassay. The parents answered questions regarding...

  15. Collective Functionality through Bacterial Individuality

    Science.gov (United States)

    Ackermann, Martin

    According to the conventional view, the properties of an organism are a product of nature and nurture - of its genes and the environment it lives in. Recent experiments with unicellular organisms have challenged this view: several molecular mechanisms generate phenotypic variation independently of environmental signals, leading to variation in clonal groups. My presentation will focus on the causes and consequences of this microbial individuality. Using examples from bacterial genetic model systems, I will first discuss different molecular and cellular mechanisms that give rise to bacterial individuality. Then, I will discuss the consequences of individuality, and focus on how phenotypic variation in clonal populations of bacteria can promote interactions between individuals, lead to the division of labor, and allow clonal groups of bacteria to cope with environmental uncertainty. Variation between individuals thus provides clonal groups with collective functionality.

  16. Prevention of bacterial adhesion

    DEFF Research Database (Denmark)

    Klemm, Per; Vejborg, Rebecca Munk; Hancock, Viktoria

    2010-01-01

    that imposes selection pressure for resistant bacteria. New approaches are urgently needed. Targeting bacterial virulence functions directly is an attractive alternative. An obvious target is bacterial adhesion. Bacterial adhesion to surfaces is the first step in colonization, invasion, and biofilm formation....... As such, adhesion represents the Achilles heel of crucial pathogenic functions. It follows that interference with adhesion can reduce bacterial virulence. Here, we illustrate this important topic with examples of techniques being developed that can inhibit bacterial adhesion. Some of these will become...

  17. Vancomycin prophylaxis of experimental Streptococcus sanguis. Inhibition of bacterial adherence rather than bacterial killing.

    OpenAIRE

    Bernard, J. P.; Francioli, P.; Glauser, M P

    1981-01-01

    Using a strain of Streptococcus sanguis tolerant to vancomycin to infect aortic vegetations in rats, we found that prophylactic intravenous vancomycin given 30 min before bacterial challenge decreased the incidence of endocarditis from 88 to 8% (P less than 10(-5)). Because peak vancomycin serum levels were below the minimal bactericidal concentration, mechanisms of protection other than bacterial killing were investigated. S. sanguis were incubated with inhibitory concentration of vancomycin...

  18. Exposure to airborne microorganisms, dust and endotoxin during flax scutching on farms.

    Science.gov (United States)

    Krysińska-Traczyk, Ewa; Skórska, Czesława; Prazmo, Zofia; Sitkowska, Jolanta; Cholewa, Grazyna; Dutkiewicz, Jacek

    2004-01-01

    Microbiological air sampling was performed on 5 flax farms located in eastern Poland. Air samples for determination of the concentrations of microorganisms, dust and endotoxin were collected in barns during machine scutching of flax stems by the farmers. The concentrations of mesophilic bacteria ranged from 203.5-698.8 x 10(3) cfu/m3, of Gram-negative bacteria from 27.2-123.4 x 10(3) cfu/m3, of thermophilic actinomycetes from 0.5-2.6 x 10(3) cfu/m3, and of fungi from 23.4-99.8 x 10(3) cfu/m3. The concentrations of total airborne microorganisms (bacteria + fungi) were within a range of 245.0-741.0 x 10(3) cfu/m3. The values of the respirable fraction of total airborne microflora on the examined farms were between 45.5-98.3%. Corynebacteria (irregular Gram-positive rods, mostly Corynebacterium spp.) were dominant at all sampling sites, forming 46.8-67.8% of the total airborne microflora. Among Gram-negative bacteria, the most numerous species was Pantoea agglomerans (synonyms: Erwinia herbicola, Enterobacter agglomerans), known to have strong endotoxic and allergenic properties. Among fungi, the allergenic species Alternaria alternata prevailed. Altogether, 25 species or genera of bacteria and 10 species or genera of fungi were identified in the farm air during flax scutching; of these, 11 and 6 species or genera respectively were reported as having allergenic and/or immunotoxic properties. The concentrations of airborne dust ranged within 43.7-648.1 mg/m3 (median 93.6 mg/m3), exceeding on all farms the Polish OEL value of 4 mg/m3. The concentrations of airborne endotoxin ranged within 16.9-172.1 microg/m3 (median 30.0 microg/m3), exceeding at all sampling sites the suggested OEL value of 0.2 microg/m). In conclusion, flax farmers performing machine scutching of flax could be exposed to large concentrations of airborne microorganisms, dust and endotoxin, posing a risk of work-related respiratory disease. PMID:15627342

  19. Bacterial and fungal markers in tobacco smoke

    Energy Technology Data Exchange (ETDEWEB)

    Szponar, B., E-mail: szponar@iitd.pan.wroc.pl [Lund University, Dept. of Laboratory Medicine, Soelvegatan 23, 223 62 Lund (Sweden); Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Rudolfa Weigla 12, 53-114 Wroclaw (Poland); Pehrson, C.; Larsson, L. [Lund University, Dept. of Laboratory Medicine, Soelvegatan 23, 223 62 Lund (Sweden)

    2012-11-01

    Previous research has demonstrated that cigarette smoke contains bacterial and fungal components including lipopolysaccharide (LPS) and ergosterol. In the present study we used gas chromatography-mass spectrometry to analyze tobacco as well as mainstream and second hand smoke for 3-hydroxy fatty acids (3-OH FAs) of 10 to 18 carbon chain lengths, used as LPS markers, and ergosterol, used as a marker of fungal biomass. The air concentrations of LPS were 0.0017 nmol/m{sup 3} (N = 5) and 0.0007/m{sup 3} (N = 6) in the smoking vs. non-smoking rooms (p = 0.0559) of the studied private houses, and 0.0231 nmol/m{sup 3} (N = 5) vs. 0.0006 nmol/m{sup 3} (N = 5) (p = 0.0173), respectively, at the worksite. The air concentrations of ergosterol were also significantly higher in rooms with ongoing smoking than in rooms without smoking. A positive correlation was found between LPS and ergosterol in rooms with smoking but not in rooms without smoking. 3-OH C14:0 was the main 3-OH FA, followed by 3-OH C12:0, both in mainstream and second hand smoke and in phenol:water smoke extracts prepared in order to purify the LPS. The Limulus activity of the phenolic phase of tobacco was 3900 endotoxin units (EU)/cigarette; the corresponding amount of the smoke, collected on filters from 8 puffs, was 4 EU/cigarette. Tobacco smoking has been associated with a range of inflammatory airway conditions including COPD, asthma, bronchitis, alveolar hypersensitivity etc. Significant levels of LPS and ergosterol were identified in tobacco smoke and these observations support the hypothesis that microbial components of tobacco smoke contribute to inflammation and airway disease. -- Highlights: Black-Right-Pointing-Pointer Air concentration of bacterial and fungal markers is significantly higher in rooms with ongoing smoking than without smoking. Black-Right-Pointing-Pointer Bacterial LPS correlates with fungal marker in rooms with ongoing smoking but not without smoking. Black-Right-Pointing-Pointer LPS

  1. Effect of Dachengqi Granules(大承气汤颗粒剂)on Level of Endotoxin in Enterogenous Endotoxemia in Rats

    Institute of Scientific and Technical Information of China (English)

    林秀珍; 郑尧; 马德禄; 谢文利; 赵珊; 吴咸中

    2002-01-01

    Objective:To study the effect of Dachengqi Granule (DCQG) on endotoxin level in rats ofenterogenous endotoxemia. Methods: Enterogenous endotoxemia model of rats was made by peritoneal in-jection of E. coli. In the treated groups, the rats were treated with different doses of DCQG, cefaclor andDCQG plus cefaclor respectively, starting from 3 days before to 1 day after modeling. Endotoxin levels inplasma, urine and stool were determined and compared. Results: Except for the cefaclor group, the plasmaand urine endotoxin levels in the treated groups were lower significantly (P<0.05), while the endotoxinlevel in stool was obviously higher than that in the untreated model group. Conclusion: DCQG has the intes-tinal barrier protective effect including anti-bacteria, anti-inflammation and clearing endotoxin.

  2. Effects of endotoxin on endothelin receptor in hepatic and intestinal tissues after endotoxemia in rats

    Institute of Scientific and Technical Information of China (English)

    Bao Hua Liu; Hui Sun Chen; Ji Hong Zhou; Nan Xiao

    2000-01-01

    @@INTRODUCTION Endothelins (ETs) has a potent and sustained vasoconstrictive effect on a variety of blood vessels.The vascular smooth muscle cell (VSMC) is the target for ETs. VSMC of the whole body contains endothelin receptor (ETR)[1]. A great number of experiments have shown that three distinct complementary DNAs of ETR have been identified i. e., endothelin A receptor (ETA receptor),endothelin B receptor ( ETB receptor ) and endothelin C receptor (ETc receptor). ETA receptor was expressed in VSMC responsible for the contraction[2]. The aim of this study is to confirm the effects of endotoxin on the activity of ETR, and the transcription and expression of ETA receptor mRNA in hepatic and intestinal tissues.

  3. Thrombin generation by exposure of blood to endotoxin: a simple model to study disseminated intravascular coagulation.

    Science.gov (United States)

    Stief, T W

    2006-04-01

    Pathologic disseminated intravascular coagulation (PDIC) is a serious complication in sepsis. In an in-vitro system consisting of incubation of fresh citrated blood with lipopolysaccharides (LPS) or glucans and subsequent plasma recalcification plasmatic thrombin was quantified. Five hundred microliters of freshly drawn citrated blood of healthy donors were incubated with up to 800 ng/mL LPS (Escherichia coli) or up to 80 microg/mL Zymosan A (ZyA; Candida albicans) for 30 minutes at room temperature (RT). The samples were centrifuged, and 30 microL plasma were recalcified with 1 volume or less of CaCl(2) (25 micromoles Ca(2+)/mL plasma). After 0 to 12 minutes (37 degrees C), 20 microL 2.5 M arginine, pH 8.6, were added. Thirty microliters 0.9 mM HD-CHG-Ala-Arg-pNA in 2.3 M arginine were added, and the absorbance increase at 405 nm was determined. Fifty microliters plasma were also incubated with 5 microL 250 mM CaCl2 for 5, 10, or 15 minutes (37 degrees C). Fifty microliters 2.5 M arginine stops coagulation, and 50 microL 0.77 mM HD-CHG-Ala-Arg-pNA in 2.3 M arginine starts the thrombin detection. The standard was 1 IU/mL thrombin in 7% human albumin instead of plasma. Arginine was also added in the endotoxin exposure time (EET) or in the plasma coagulation reaction time (CRT). Tissue factor (TF)-antigen and soluble CD14 were determined. LPS at blood concentrations greater than 10 ng/mL or ZyA at greater than 1 microg/mL severalfold enhance thrombin generation, when the respective plasmas are recalcified. After 30 minutes EET at RT, the thrombin activity at 12 minutes CRT generated by the addition of 200 ng/mL LPS or 20 microg/mL ZyA is approximately 200 mIU/mL compared to approximately 20 mIU/mL without addition of endotoxin, or compared to about 7 mIU/mL thrombin at 0 minutes CRT. Arginine added to blood or to plasma inhibits thrombin generation; the inhibitory concentration 50% (IC 50) is approximately 15 mM plasma concentration. Endotoxin incubation of blood

  4. 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.

  5. Association between dwelling characteristics and concentrations of bacteria, endotoxin and fungi in settling dust

    DEFF Research Database (Denmark)

    Spilak, Michal; Knudsen, Sofie; Madsen, Anne Mette;

    2014-01-01

    Numerous studies indicate that the presence of fungi, bacteria and endotoxins in the indoor environment is strongly associated with serious health risks such as asthma, allergies and respiratory discomfort (Horick et al. 2005, Denning et al. 2006, Tischer et al. 2011). Hence there is a need...... to determine potential predictors that influence the presence of these microorganisms in homes. Fungi can grow on different indoor materials such as humid gypsum-boards. Fungal spores can upon exposure to airflow be aerosolised (Madsen et al. 2006). The size of the fungal spores in indoor air is usually below...... 5 μm (Chang and Gershwin, 2004). Fungi are naturally present outdoors and levels of outdoor fungi are affected by local climate conditions, such as rainfall and temperature (Chang and Gershwin, 2004). Outdoor fungi contribute to the indoor level of fungi via infiltration through the building...

  6. [Effect of antihypoxant infusion on microbial endotoxin circulation in obstructive jaundice patients].

    Science.gov (United States)

    Iakovlev, A Ia; Niazmatov, A A; Zarechnova, N V; Zaĭtsev, R R; Emel'ianov, N V; Mokrov, K V; Chichkanova, A S

    2013-01-01

    The dynamics of microbial endotoxemia in group of 149 patients with cholelithiasis complicated by obstructive jaundice has been studied as dependent on the perioperative tactics of infusion therapy. The perioperative period in obstructive jaundice patients is complicated by a significant increase in lipopolysaccharidemia caused by translocation mechanisms and disorders of the liver detoxification function. In Group 1, 47 patients received infusion therapy including Ringer's solution and 10% glucose solution at a 1:1 ratio. In group 2, 55 patients received infusion therapy with only Sterofundin G-5 solution. In Group 3, 47 patients received the infusion therapy with remaxol in a dose of 800 ml/day. It is established that the infusion of Sterofundin G-5, and to a greater extent the infusion of remaxol, reduces the early postoperative degree of decompensation and accelerates recovery of the detoxifying function of Kupffer cells with respect to microbial endotoxin. PMID:23631281

  7. Effects of the immunomodulator, VGX-1027, in endotoxin-induced uveitis in Lewis ratsTumor necrosis factor-alpha binding capacity and anti-infliximab antibodies measured by fluid-phase radioimmunoassays as predictors of clinical efficacy of infliximab in Crohn's disease

    DEFF Research Database (Denmark)

    Mangano, K.; Sardesai, N.Y.; Quattrocchi, C.;

    2008-01-01

    . Here, we have studied the effects of VGX-1027 on the development of endotoxin-induced uveitis (EIU) in male Lewis rats, as a model of inflammatory ocular diseases in humans. EXPERIMENTAL APPROACH: EIU was induced by a single footpad injection of 200 microg lipopolysaccharide (LPS). Groups of rats were......, immunological and histological signs of EIU. KEY RESULTS: The rats treated with VGX-1027 within 6 h after LPS challenge exhibited milder clinical, histological and laboratory signs of EIU than those treated with vehicle. CONCLUSION AND IMPLICATIONS: This study provides the first evidence that systemic treatment...

  8. Effect of bacterial lipopolysaccharide on gastric emptying of liquids in rats

    Directory of Open Access Journals (Sweden)

    E.F. Collares

    1997-02-01

    Full Text Available The objectives of the present investigation were 1 to study the effect of bacterial lipopolysaccharide (LPS on rat gastric emptying (GE and 2 to investigate a possible involvement of the vagus nerve in the gastric action of LPS. Endotoxin from E. coli (strain 055:B5 was administered sc, ip or iv to male Wistar rats (220-280 g body weight at a maximum dose of 50 µg/kg animal weight. Control animals received an equivalent volume of sterile saline solution. At a given time period after LPS administration, GE was evaluated by measuring gastric retention 10 min after the orogastric infusion of a test meal (2 ml/100 g animal weight, which consisted of 0.9% NaCl plus the marker phenol red (6 mg/dl. One group of animals was subjected to bilateral subdiaphragmatic vagotomy or sham operation 15 days before the test. A significant delay in GE of the test meal was observed 5 h after iv administration of the endotoxin at the dose of 50 µg/kg animal weight. The LPS-induced delay of GE was detected as early as 30 min and up to 8 h after endotoxin administration. The use of different doses of LPS ranging from 5 to 50 µg/kg animal weight showed that the alteration of GE was dose dependent. In addition, vagotomized animals receiving LPS displayed a GE that was not significantly different from that of the sham control group. However, a participation of the vagus nerve in LPS-induced delay in GE could not be clearly demonstrated by these experiments since vagotomy itself induced changes in this gastric parameter. The present study provides a suitable model for identifying the mechanisms underlying the effects of LPS on gastric emptying

  9. [Effect of xuebijing oral effervescent tablet on endotoxin induced fever and disseminated intravascular coagulation rabbit model].

    Science.gov (United States)

    Guo, Shan-Shan; Gao, Ying-Jie; Tian, Xue-Chuan; Jin, Ya-Hong; Liu, Fang-Zhou; Cui, Xiao-Lan

    2013-08-01

    In order to discover the mechanism of Xuebijing oral effervescent tablet (XBJOET) to treat infectious diseases, the effect of XBJOET on endotoxin induced rabbit fever and disseminated intravascular coagulation (DIC) was investigated. Auricle microcirculation in rabbit was detected by laser speckle blood perfusion imager system; coagulation function was measured by coagulation analyzer, fibrinolytic system was quantified by Elisa assay and micro thrombosis in tissues was observed with HE staining under light microscope. The results demonstrated that the body temperature of rabbit decreased significantly at 1-3 h after administration with 4.8, 2.4 and 1.2 g x kg(-1) XBJOET to endotoxin induced DIC rabbit model, the auricle microcirculation blood flow in model group (54.45 +/- 14.53) PU was lower than that in control group (77.18 +/- 12.32) PU. The auricle microcirculation blood flow increased markedly and there was significant difference between model group and 1.2 g x kg(-1) XBJOET group. There was significant difference between model group and control group in the content of PAI1 and FIB. The PAI1 levels in model and control groups were (30.48 +/- 2.46) ng x mL(-1) and (20.93 +/- 3.25) ng x mL(-1), respectively. The FIB levels in model and control group were (3.34 +/- 1.09) g x L(-1) and (4.84 +/- 1.10) g x L(-1), respectively. The content of PAI1 in rabbit plasma decreased notably, there were significant differences between model group and 4.8, 2.4 g x kg(-1) XBJOET groups. On the contrary the content of FIB increased. XBJOET possessed pharmacological activities of curing infectious fever and DIC, the mechanism of which is related to amelioration of microcirculation disturbance, inhibition of fibrinolytic system activation and coagulation and micro thrombosis elimination.

  10. Glycyrrhizin attenuates endotoxin- induced acute liver injury after partial hepatectomy in rats

    Directory of Open Access Journals (Sweden)

    B. Tang

    2007-12-01

    Full Text Available Massive hepatectomy associated with infection induces liver dysfunction, or even multiple organ failure and death. Glycyrrhizin has been shown to exhibit anti-oxidant and anti-inflammatory activities. The aim of the present study was to investigate whether glycyrrhizin could attenuate endotoxin-induced acute liver injury after partial hepatectomy. Male Wistar rats (6 to 8 weeks old, weighing 200-250 g were randomly assigned to three groups of 24 rats each: sham, saline and glycyrrhizin. Rats were injected intravenously with lipopolysaccharide (LPS 24 h after 70% hepatectomy. Glycyrrhizin, pre-administered three times with 24 h intervals 48 h before hepatectomy, prolonged the survival of rats submitted to partial hepatectomy and LPS injection, compared with saline controls. Glycyrrhizin was shown to attenuate histological hepatic changes and significantly reduced serum levels of aspartate aminotransferase, alanine aminotransferase, and lactic dehydrogenase, at all the indicated times (6 rats from each were sacrificed 1, 3, 6, and 9 h after LPS injection, compared with saline controls. Glycyrrhizin also significantly inhibited hepatocyte apoptosis by down-regulating the expression of caspase-3 and inhibiting the release of cytochrome C from mitochondria into the cytoplasm. The anti-inflammatory activity of glycyrrhizin may rely on the inhibition of release of tumor necrosis factor-a, myeloperoxidase activity, and translocation of nuclear factor-kappa B into the nuclei. Glycyrrhizin also up-regulated the expression of proliferating cell nuclear antigen, implying that it might be able to promote regeneration of livers harmed by LPS. In summary, glycyrrhizin may represent a potent drug protecting the liver against endotoxin-induced injury, especially after massive hepatectomy.

  11. Exposure to particulates, microorganisms, beta(1-3)-glucans, and endotoxins during soybean harvesting.

    Science.gov (United States)

    Roy, Chad J; Thorne, Peter S

    2003-01-01

    Sclerotinia sclerotiorum is an emerging fungal pathogen affecting soybeans in the United States. In response to its emergence, exposures to particulates, bioaerosols, endotoxins, S. sclerotiorum, and beta(1-3)-glucans were characterized during soybean harvesting. Air sampling was performed on soybean harvesters (combines) and on the farmers in closed cabs as personal samples during harvesting at 17 farms in 1997 and repeated at 15 in 1998. S. sclerotiorum infestation was evident in the fields at 8 of the sites (44%). The geometric mean concentrations (and geometric standard deviations) measured on the combines in 1998 were as follows: total dust, 11.9 (2.8) mg/m(3); inhalable dust 11.7 (6.4) mg/m(3); and beta(1-3)-glucans, 5027 (7) ng/m(3). Values for the personal samples in 1998 were as follows: total dust, 1.2 (6.7) mg/m(3); inhalable dust, 1.1 (5.3); and beta(1-3)-glucans, 674 (9) ng/m(3). These concentrations were two- to threefold higher than in the previous year. Ambient endotoxin concentrations were 4438 EU/m(3) in Year I and 459 EU/m(3) in Year II. Particle size distribution measurements on the combines yielded mass median aerodynamic diameters of 6.6 microm on the combine and 4.0 microm inside the combine cab. Closed combine cabs provided an average workplace protection factor of 11.7 for total dust. Nevertheless, personal exposures to organisms inside combine cabins ranged from 3.6 x 10(4) to 4.0 x 10(8) organisms/m(3). These data indicate the potential exists for high exposures to organic dust and bioaerosols during soybean harvesting. PMID:12908864

  12. The Level of Vasopressin is not solely resulted from the Concentration of Endotoxin but Proportional to Creatinine in Dogs with Pyometra

    OpenAIRE

    W. Y. Shia1, K. C. Tung1, 2, S. C. Chang1, 2, C. H. Yang1, 2, C. H. Lee2, C. C. Chou1 and W. M. Lee1, 2*

    2011-01-01

    Pyometra is one of the most common reproductive disorder characterized as fluid accumulation mainly pus and Gram-negative bacteria isolated from uterus in bitches. Impaired function of vasopressin is found in the disease. The impact of the circulating endotoxin concentration on vasopressin involved water regulation is still unclear. To document the effect of endotoxin on vasopressin-involved water regulation in dogs with pyometra, blood samples were collected for examination of circulating en...

  13. Effect of endotoxin on the celluar activity and secretion of endothelin-1 in cultured human umbilical vein endothelial cells

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    AIM and METHODS: To investigate the effect of endotoxin on the celluar activity and secretion of endothelin-1 by radioimmunoassay and MTT methods in cultured human umbilical vein endothelial cells stimulated by E coli endotoxin (E coli O55:B5, Sigma) of various concentrations (1 g/L, 100 mg/L, 10 mg/L,1 mg/L,100 μg/L,10 μg/L, 1 μg/L) and at the same time interval (HUVEC stimulated by endotoxin for 6 hours) in vitro.RESULTS:Endotoxin showed a slightly inhibitory effect on the viability of endothelial cells at low doses (1 μg/L, 10 μg/L, 100 μg/L, 1 mg/L). The viabilities were 92.00%±1.45%, 91.81%±2.03%, 89.52%±1.49%, 88.35%±1.88%, respectively, versus control group, P<0.01. The cells were impaired significantly at the higher dose of LPS (100 mg/L), the viability was 80.49%±8.76%, versus control group, P<0.01. The cells were killed evidently at the concentration of LPS (1 g/L), the viability was 73%±8%, versus control group, P<0.01. The secretion of ET-1 increased gradually with the concentration of endotoxin manifolding. The concentration of ET-1 reached its peak at the dose of 100 μg/L, and it was (324.384±17.023) ng/L, versus control group (251.636±17.023) ng/L, P<0.01. Endotoxin was effective in stimulating the endothelial cells to secret ET-1 in a dose dependent manner. CONCLUSION: These findings suggested ET-1 may be one of the important factors in endotoxic shock, and the increase in plasma ET-1 level in endotoxemia may be associated with increase in ET-1 secretion.

  14. Hypo-responsiveness of interleukin-8 production in human embryonic epithelial intestine 407 cells independent of NF-κB pathway: New lessons from endotoxin and ribotoxic deoxynivalenol

    International Nuclear Information System (INIS)

    Mucosal epithelium senses external toxic insults and transmits the danger signals into the epithelial cells in order to activate a broad range of inflammatory responses. However, pre-exposure to the commensal endotoxins can induce inflammatory tolerance and maintain the homeostasis without excessive immune responses. We recently reported that ribotoxin deoxynivalenol (DON) and its derivatives elicited the pro-inflammatory response as the mucosal insults in human epithelial cells. Taking the knowledge into consideration, we tested the hypothesis that endotoxin pre-exposure can attenuate ribotoxin-induced epithelial interleukin-8 (IL-8) production via a tolerance mechanism. Pre-exposure to endotoxin repressed IL-8 release and its gene expression. However, inflammatory tolerance was not mediated by the attenuated NF-κB activation which has been generally recognized as the major mediator of LPS-mediated toll-like receptor (TLR) signaling pathway. Instead, pre-exposure to endotoxin was observed to trigger the delayed induction of peroxisome proliferator-activated receptor gamma (PPAR-γ) which contributed to the diminished IL-8 production in the human epithelial cells. Moreover, endogenous PPAR-γ agonist suppressed toxicant-mediated interleukin-8 production and IL-8 mRNA stability. Taken together, endotoxin induced hypo-production of pro-inflammatory cytokine IL-8 in the human epithelial cells, which was associated with the delayed activation of PPAR-γ expression by pre-existing endotoxin

  15. The effects of cage design on airborne allergens and endotoxin in animal rooms: high-volume measurements with an ion-charging device.

    Science.gov (United States)

    Platts-Mills, James; Custis, Natalie; Kenney, Alice; Tsay, Amy; Chapman, Martin; Feldman, Sanford; Platts-Mills, Thomas

    2005-03-01

    Respiratory symptoms related to both endotoxins and animal allergens continue to be an important cause of occupational disease for animal technicians and scientists working with rodents. Better sampling methods for airborne allergens and endotoxin are needed to help standardize compliance with federal occupational health regulations. Using an ion-charging device, we sampled 20 mouse rooms and four rat rooms at the University of Virginia, along with 43 domestic living rooms in houses in the Charlottesville area with at least one cat or dog. The use of filter tops on cages corresponds to a 50-fold reduction in mean levels of both airborne allergens (P < 0.001) and endotoxin (P < 0.001). The use of vented cages with filtered exhaust ports was associated with additional reductions. However, the mean airborne endotoxin level in all rooms using filter tops without a filtered exhaust port on the cages was significantly lower (P = 0.003) than the level in domestic living rooms. Our results for maximum airborne allergens or endotoxin are comparable with previous reports. However, the sensitivity of the technique allows an accurate assessment of low-level exposure, which makes it possible to evaluate the effect of cage designs. In addition, this approach allows direct comparison with results for airborne allergen and endotoxin in domestic homes. The results could allow a more consistent approach to the application of occupational health guidelines. PMID:15773770

  16. Déjà-vu from the nineties: is there a perspective for anti-endotoxin strategies to improve the outcome of multiple trauma patients?

    Science.gov (United States)

    Menger, Michael D.

    2016-01-01

    A recent cohort study of Charbonney et al. indicates that multiple trauma patients develop endotoxemia also in the absence of Gram-negative infection. This is most probably due to an increase of gut permeability. Non-survivors as well as patients with cardiovascular dysfunction and multiple organ failure (MOF) show significantly higher endotoxin levels at 24 h after injury compared to survivors and patients without MOF. These results are like a déjà-vu from the nineties of the last century, where several studies reported endotoxemia during the initial 24 h after multiple trauma with development of MOF and death at endotoxin levels >10 and >12 pg/mL, respectively. Of interest, other multiple trauma patient studies in the nineties have shown endogenous anti-endotoxin antibody production in survivors and reduced antibody production in non-survivors, which died from MOF. Although all these studies have pointed towards a mechanistic role of endotoxin in the fatal outcome after multiple injuries, clinical anti-endotoxin studies are still lacking. Thus, the future perspective must be prospective randomized multicenter trials, which have to elucidate the capability of anti-endotoxin treatment strategies to improve outcome in multiple trauma patients. PMID:27620806

  17. A comparison of haemocoagulation tests in the experimental endotoxin model DIC and in rats whole-body irradiated by 250 Gy

    International Nuclear Information System (INIS)

    A comparison of results of tests performed with the endotoxin model of disseminated intravascular coagulation (DIC) and with irradiated groups of rats led to the conclusion that after whole-body irradiation with the high dose of 250 Gy, DIC occurs, in spite of the fact that the first stage, the hypercoagulation condition, can hardly be observed. In the experimental endotoxin model, an increase of activated partial thromboplastin test (APTT) values and prolongation of the thrombin time was observed up to 24 hours for two endotoxin doses. After both endotoxin doses, the fibrinogen level was transiently decreased with a subsequent increase. The fibrin monomers correspond to a decrease in the fibrinogen level. After the first dose, they were positive between the 3rd and 12th hours, and after the second dose, positivity was observed 6 hours after the application. The antithrombin III level was decreased after 12 hours for both endotoxin doses. The thrombocyte count was considerably reduced already from the 6th hour after administering endotoxin to the end of the experiment. Considerable changes in thrombocyte aggregation were observed only 3 hours after administering the second dose. When comparing the resulting values of these tests with values observed in irradiated animals, a certain agreement was found in the nature of the changes after exposure to 250 Gy. The fibrinogen level was transiently decreased 3 hours after irradiation, when considerable changes in the thrombocyte aggregation also occurred. (author) 5 figs., 17 refs

  18. Emerging bacterial pathogens: the past and beyond.

    Science.gov (United States)

    Vouga, M; Greub, G

    2016-01-01

    Since the 1950s, medical communities have been facing with emerging and reemerging infectious diseases, and emerging pathogens are now considered to be a major microbiologic public health threat. In this review, we focus on bacterial emerging diseases and explore factors involved in their emergence as well as future challenges. We identified 26 major emerging and reemerging infectious diseases of bacterial origin; most of them originated either from an animal and are considered to be zoonoses or from water sources. Major contributing factors in the emergence of these bacterial infections are: (1) development of new diagnostic tools, such as improvements in culture methods, development of molecular techniques and implementation of mass spectrometry in microbiology; (2) increase in human exposure to bacterial pathogens as a result of sociodemographic and environmental changes; and (3) emergence of more virulent bacterial strains and opportunistic infections, especially affecting immunocompromised populations. A precise definition of their implications in human disease is challenging and requires the comprehensive integration of microbiological, clinical and epidemiologic aspects as well as the use of experimental models. It is now urgent to allocate financial resources to gather international data to provide a better understanding of the clinical relevance of these waterborne and zoonotic emerging diseases.

  19. Susceptibility of legume pod borer (LPB), Maruca vitrata to delta-endotoxins of Bacillus thuringiensis (Bt) in Taiwan.

    Science.gov (United States)

    Srinivasan, R

    2008-01-01

    Baseline susceptibility of legume pod borer (LPB) to the insecticidal crystal proteins (ICPs) from Bacillus thuringiensis, viz, Cry1Aa, Cry1Ab, Cry1Ac, Cry1Ca and Cry2Aa was assessed in Taiwan. Insect bioassays were performed by incorporating the Bt delta-endotoxins into the LPB artificial diet. The efficacy of different Bt delta-endotoxins against second instar larvae of LPB showed that the toxin Cry1Ab was the most potent toxin (LC(50) 0.207ppm), followed by Cry1Ca, Cry1Aa, Cry2Aa and Cry1Ac in descending order, with LC(50)s 0.477ppm, 0.812ppm, 1.058ppm and 1.666ppm, respectively. Hence, Cry1Ab and/or Cry1Ca toxins would provide effective control of early larval stages of LPB.

  20. Glucocorticosteroids: as Adjuvant Therapy for Bacterial Infections

    Directory of Open Access Journals (Sweden)

    WONDIM MELKAM

    2015-01-01

    Full Text Available Glucocorticoids (GCs, synthetic analogues of the natural steroid hormones, are well known for their antiinflammatory and immunosuppressive properties in the periphery. They are widely and successfully used in the treatment of autoimmune diseases, chronic inflammation, and transplant rejection. Nowadays, GCs are claimed to have a beneficial role being as adjunct therapy in various infections. Different studies have been conducted to investigate their use as adjuvant therapy for different bacterial infection. This review, therefore, summarizes various bacterial infections for which glucocorticoids are reported to be used as adjuvant therapy, strategies for administration of glucocorticoids, and challenges of using glucocorticoids as adjuvant therapy.

  1. Comparative Study of Plasma Endotoxin with Procalcitonin Levels in Diagnosis of Bacteremia in Intensive Care Unit Patients

    OpenAIRE

    Tao Wang; Yun-Liang Cui; Zhao-Fen Lin; De-Chang Chen

    2016-01-01

    Background: Both procalcitonin (PCT) and plasma endotoxin levels cannot be solely used for a definite diagnosis of bacteremia or sepsis, and there has been few study comparing the values of the two biomarkers for the diagnosis of bacteremia. The aim of this study was to identify bacteria causing bacteremia and evaluate the role of the two biomarkers in the diagnosis of bacteremia in Intensive Care Unit (ICU). Methods: The medical records of 420 patients in ICU were retrospectively reviewed...

  2. Comparative Study of Plasma Endotoxin with Procalcitonin Levels in Diagnosis of Bacteremia in Intensive Care Unit Patients

    OpenAIRE

    WANG, Tao; Cui, Yun-Liang; Lin, Zhao-fen; Chen, De-Chang

    2016-01-01

    Background: Both procalcitonin (PCT) and plasma endotoxin levels cannot be solely used for a definite diagnosis of bacteremia or sepsis, and there has been few study comparing the values of the two biomarkers for the diagnosis of bacteremia. The aim of this study was to identify bacteria causing bacteremia and evaluate the role of the two biomarkers in the diagnosis of bacteremia in Intensive Care Unit (ICU). Methods: The medical records of 420 patients in ICU were retrospectively reviewed. P...

  3. Mold and Endotoxin Levels in the Aftermath of Hurricane Katrina: A Pilot Project of Homes in New Orleans Undergoing Renovation

    OpenAIRE

    Chew, Ginger L.; Wilson, Jonathan; Rabito, Felicia A.; Grimsley, Faye; Iqbal, Shahed; Reponen, Tiina; Thorne, Peter S.; Dearborn, Dorr G.; Morley, Rebecca L.; Michael L Muilenberg

    2006-01-01

    Background: After Hurricane Katrina, many New Orleans homes remained flooded for weeks, promoting heavy microbial growth. Objectives: A small demonstration project was conducted November 2005–January 2006 aiming to recommend safe remediation techniques and safe levels of worker protection, and to characterize airborne mold and endotoxin throughout cleanup. Methods: Three houses with floodwater lines between 0.3 and 2 m underwent intervention, including disposal of damaged furnishings and dryw...

  4. [The effect of a soil extract on the development of Bacillus thuringiensis and on its synthesis of an insecticidal endotoxin].

    Science.gov (United States)

    Dregval', O A; Cherevach, N V; Andrienko, O E; Vinnikov, A I

    1999-01-01

    Selection of effective and inexpensive nutrient medium for cultivation of entomopathogenic bacteria Bacillus thuringiensis was carried out. The medium with molasses [correction of patoka], corn extract and mineral salts has been chosen. Addition of a soil extract to the medium enhanced growth of microorganisms, increased the rate of culture development, the yield of spore-crystalline material and quantity of synthesized endotoxin. Apparently the strengthening effect belongs to humic substances contained in the soil extract. PMID:10565149

  5. Experimentelle Untersuchungen zum Proliferationsverhalten von NSCLC-Zelllinien unter Endotoxin-Stimulation : die Rolle des Cyclooxygenase-Metabolismus

    OpenAIRE

    Linsel, Sören

    2016-01-01

    Lungenkarzinome haben weltweit eine hohe Inzidenz und Mortalität. Da pulmonale Infektionen zusätzlich zum Lungenkarzinom nicht nur die Diagnose erschweren, sondern auch die Prognose verschlechtert, sollte ein möglicher Zusammenhang zwischen Infektion und Tumorprogression in der vorliegenden Arbeit genauer betrachtet werden. Hierzu wurden Versuche durchgeführt, bei denen drei NSCLC-Zelllinien (A549, H1299, H226) mit verschiedenen Konzentrationen des Endotoxins LPS, Membranbestandteil von gramn...

  6. Inhibition of endotoxin-induced interleukin-6 production by synthetic lipid A partial structures in human peripheral blood mononuclear cells.

    OpenAIRE

    Wang, M. H.; Flad, H D; Feist, W; Brade, H; Kusumoto, S; Rietschel, E T; Ulmer, A J

    1991-01-01

    The effect of two synthetic lipid A partial structures, compound 406 (or LA-14-PP, identical in structure to the lipid A precursor, known as Ia or IVa) and compound 401 (lipid X), on the in vitro modulation of endotoxin (lipopolysaccharide)-induced interleukin-6 production by human blood mononuclear cells was investigated. Lipopolysaccharide of Salmonella abortus equi and synthetic Escherichia coli-type lipid A (compound 506, or LA-15-PP) had potent interleukin-6-inducing capacities. The maxi...

  7. Monitoring the effectiveness of root canal procedures on endotoxin levels found in teeth with chronic apical periodontitis

    OpenAIRE

    MARINHO, Ariane Cassia Salustiano; MARTINHO, Frederico Canato; ZAIA, Alexandre Augusto; Ferraz, Caio Cezar Randi; GOMES, Brenda Paula Figueiredo de Almeida

    2014-01-01

    Objective: The aim of this study was to monitor the effectiveness of root canal procedures by using different irrigants and intracanal medication on endotoxin levels found in root canals of teeth with chronic apical periodontitis. Material and Methods: Thirty root canals of teeth with pulpal necrosis associated with periapical lesions were selected and randomly divided into groups according to the irrigants used: GI - 2.5% NaOCl, GII - 2% chlorhexidine (CHX) gel, and GIII - saline solution ...

  8. Are bovine teeth a suitable substitute for human teeth in in vitro studies to assess endotoxin load in root canals?

    Directory of Open Access Journals (Sweden)

    Tiago André Fontoura de MELO

    2015-01-01

    Full Text Available The present study aimed to determine the feasibility of using bovine teeth as a suitable alternative for human teeth, in experiments involving in vitro endotoxin contamination. Twenty bovine central incisors and 20 human single-root premolars had their dental crowns removed and root lengths set at 16 mm. Root canals were prepared up to #60 K-file size and sterilized with cobalt-60 gamma irradiation (20 kGy, 6 h. The teeth were randomly divided into four groups: G1-bovine teeth (bovine negative control, n = 10, G2-human teeth (human negative control, n = 10, G3-bovine teeth, inoculated withEscherichia coli (055:B55 LPS, and G4-human teeth inoculated with E. coli LPS. The G1 and G2 groups were exposed to apyrogenic water. After the teeth had been incubated at 37 °C and atmospheric humidity for 24 h, the samples of solutions in the main canals were collected with apyrogenic absorbent paper tips. LPS levels were quantified using Limulus Amebocyte Lysate assay. The data obtained were statistically analyzed using one-way ANOVA, with a significance level of 5%. A high amount of endotoxin was detected in the inoculated human teeth (G4 when compared to the sterilized teeth (G2, as well as in the inoculated bovine teeth (G3 when compared to the inoculated human teeth (G4. However, there was no statistical difference between bovine teeth before and after the E. coli endotoxin inoculation. Therefore, under the mentioned experimental conditions, the use of bovine teeth should not be a choice for laboratory research on endotoxin contamination.

  9. Profiling of 3-hydroxy fatty acids as environmental markers of endotoxin using liquid chromatography coupled to tandem mass spectrometry.

    Science.gov (United States)

    Uhlig, Silvio; Negård, Mariell; Heldal, Kari K; Straumfors, Anne; Madsø, Lene; Bakke, Berit; Eduard, Wijnand

    2016-02-19

    3-Hydroxy acids are constituents of the lipid A part of lipopolysaccharides and may potentially be used as chemical markers of endotoxin. While commercial enzymatic assays, such as the widely used Limulus amebocyte lysate (LAL) assay, commonly detect merely the water-soluble fraction of the bioactive endotoxin, the chemical approach aims to estimate the total amount of endotoxin present in a sample. Our objective was to develop a simple method for quantitative profiling of 3-hydroxy fatty acids in occupational and environmental samples based on detection with HPLC-MS/MS. We included eleven 3-hydroxy fatty acids (3-hydroxyoctanoic acid to 3-hydroxyoctadecanoic acid) in the HPLC-MS/MS based method, which involved base hydrolysis of filter samples using 1M sodium hydroxide and removal of the base as well as concentration of the fatty acids using solid-phase extraction on a functionalized polystyrene-divinylbenzene polymer. Recovery trials from spiked glass fiber filters, using threo-9,10-dihydroxyhexadecanoic acid as internal standard, gave an overall recovery of 54-86% for 3-hydroxy fatty acids of medium chain length (3-hydroxynonanoic to 3-hydroxypentadecanoic acid). 3-Hydroxyoctanoic acid and the longer chain fatty acids were more problematic yielding overall spike recoveries of 11-39%. While the 3-hydroxy fatty acid profile of pure lipopolysaccharides was dominated by 3-hydroxydecanoic, 3-hydroxydodecanoic and 3-hydroxytetradecanoic acid the aqueous phase from drilling mud contained in addition relatively high amounts of 3-hydroxyoctanoic and 3-hydroxynonanoic acid. Endotoxin activity as measured by the LAL assay was reasonably correlated (R(2)=0.54) to the sum of 3-hydroxydecanoic acid, 3-hydroxydodecanoic acid and 3-hydroxytetradecanoic acid in these samples. PMID:26818235

  10. Comments on: High fat intake leads to acute postprandial exposure to circulating endotoxin in type 2 diabetic subjects.

    Directory of Open Access Journals (Sweden)

    S Yu Vorotnikova

    2012-06-01

    Full Text Available Comments on: Harte AL, Varma MC, Tripathi G, McGee KC, Al-Daghri NM, Al-Attas OS, Sabico S, O'Hare JP, Ceriello A, Saravanan P, Kumar S, McTernan PG. High fat intake leads to acute postprandial exposure to circulating endotoxin in type 2 diabetic subjects. Diabetes Care. 2012 Feb; 35(2: 375-82

  11. Vimentin in Bacterial Infections

    Directory of Open Access Journals (Sweden)

    Tim N. Mak

    2016-04-01

    Full Text Available Despite well-studied bacterial strategies to target actin to subvert the host cell cytoskeleton, thus promoting bacterial survival, replication, and dissemination, relatively little is known about the bacterial interaction with other components of the host cell cytoskeleton, including intermediate filaments (IFs. IFs have not only roles in maintaining the structural integrity of the cell, but they are also involved in many cellular processes including cell adhesion, immune signaling, and autophagy, processes that are important in the context of bacterial infections. Here, we summarize the knowledge about the role of IFs in bacterial infections, focusing on the type III IF protein vimentin. Recent studies have revealed the involvement of vimentin in host cell defenses, acting as ligand for several pattern recognition receptors of the innate immune system. Two main aspects of bacteria-vimentin interactions are presented in this review: the role of vimentin in pathogen-binding on the cell surface and subsequent bacterial invasion and the interaction of cytosolic vimentin and intracellular pathogens with regards to innate immune signaling. Mechanistic insight is presented involving distinct bacterial virulence factors that target vimentin to subvert its function in order to change the host cell fate in the course of a bacterial infection.

  12. The effect of taurine on the relationship between NO, ADMA and homocysteine in endotoxin-mediated inflammation in HUVEC cultures.

    Science.gov (United States)

    Pasaoglu, Ozge Tugce; Turkozkan, Nurten; Ark, Mustafa; Polat, Belgin; Agilli, Mehmet; Yaman, Halil

    2014-10-01

    The aim of our study was to investigate the effect of taurine on the relationship between nitric oxide (NO), asymmetric dimethylarginine (ADMA) and homocysteine (Hcy) in endotoxin-induced human umblical vein endothelial cell (HUVEC) cultures. For this reason, four groups were formed (n=12). Control group consists of HUVEC cultures without any treatment. Lipopolysaccharide (LPS) and LPS+taurine groups were treated with 10 μg/mL endotoxin, 5 μg/mL taurine and endotoxin+taurine (same doses), respectively. Nitrite/nitrate (NOx), ADMA and Hcy levels were measured. There was a significant increase of NOx, ADMA and Hcy in endotoxemia (p<0.05). Taurine treatment elevated NOx levels significantly (p<0.01) in taurine and LPS + taurine group compared to control group, while it reduced NOx levels compared to LPS group. In contrast, taurine decreased ADMA levels to the control level both in taurine and taurine+LPS group compared to LPS. Hcy levels increased significantly compared to taurine group (p<0.05) and did not change compared to LPS group. Taurine was effective on ADMA-NO relationship whereas no beneficial effect was observed in Hcy levels (p<0.05).

  13. IL-12 suppression, enhanced endocytosis and up-regulation of MHC-Ⅱ and CD80 in dendritic cells during experimental endotoxin tolerance

    Institute of Scientific and Technical Information of China (English)

    Jing ZHANG; Jie-ming QU; Li-xian HE

    2009-01-01

    Aim: The aim of this study was to investigate endocytosis, MHC-Ⅱ expression and co-stimulatory molecule expression, as well as interleukin-12 (IL-12) production, in bone marrow dendritic cells (DCs) derived from endotoxin tolerant mice.Methods: Endotoxin tolerance was induced in C57BL/10J mice through four consecutive daily intraperitoneal injections of 1.0 mg/kg of 055:B5 Escherichia coli lipopolysaccharide (LPS). Bone marrow DCs were isolated in the presence of GM-CSF and IL-4 and purified by anti-CD11c Micro beads. FITC-dextran uptake by DCs was tested by flow cytometric analysis and the percentage of dextran-containing cells was calculated using a fluorescence microscope. The expression of surface MHC-Ⅱ, CD40, CD80, and CD86 was also detected by flow cytometric analysis. An ELISA was used for the measurement of IL-12 production by DCs with or without LPS stimulation.Results: Endotoxin tolerance was successfully induced in C57BL/10J mice, evidenced by an attenuated elevation of systemic TNF-α. DCs from endotoxin tolerant mice possessed enhanced dextran endocytosis ability. The expression of surface MHC-Ⅱ and CD80 was higher in DCs from endotoxin tolerant mice than in DCs from control mice, whereas the expression of CD40 and CD86 was not altered. Compared with DCs from normal control mice, DCs from endotoxin tolerant mice produced less IL-12 after subsequent in vitro stimulation with LPS.Conclusion: These data suggest enhanced endocytosis, selective up-regulation of MHC-Ⅱ and CD80 and IL-12 suppression in DCs during in vivo induction of endotoxin tolerance.

  14. Utilization of fodder yeast and agro-industrial by-products in production of spores and biologically - active endotoxins from Bacillus thuringiensis.

    Science.gov (United States)

    Salama, H S; Foda, M S; Selim, M H; El-Sharaby, A

    1983-01-01

    A number of newly-devised fermentation media were evaluated with respect to their ability to support sporulation and biosynthesis of endotoxins by strains of Bacillus thuringiensis that are biologically active against Spodoptera littoralis, Heliothis armigera, and Spodoptera exigua. Fodder yeast from dried cells of Saccharomyces cerevisiae could be used as a complete mono-component medium for production of highly active spore-delta-endotoxin complexes from B. thur., vars. entomocidus, kurstaki and galleriae. Highest sporulation titers were obtained at 2% fodder yeast concentration with endotoxin yields ranging between 7 to 9 grams per liter of medium. Ground horse beans and kidney bean seeds could also be used successfully as complete media for sporulation and endotoxin production. Extracts of potato tubers and sweet potato roots were efficient media for active endotoxin production from B. thur. var. kurstaki, although the obtained yields were much lower than those produced in fodder yeast media. The utilization of fish meal, cotton seed meal, and residues of chicken from the slaughter-house as media for the production of endotoxins active against Spodoptera littoralis, was not successful. On the other hand, minced citrus peels, ground seeds of dates, and wheat bran could be successfully used in combination with fodder yeast as media for production of endotoxins, active against Heliothis armigera and Spodoptera exigua. Re-utilization of culture supernatants in a second fermentation cycle after supplementation with some nutrients gave promising results with some of the strains tested. The data obtained are discussed in view of their feasibility of application. PMID:6666415

  15. Melatonin reduces bacterial translocation and apoptosis in trinitrobenzene sulphonic acid-induced colitis of rats

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    AIM:To investigate the effects of exogenous melatonin on bacterial translocation and apoptosis in a rat ulcerative colitis model.METHODS:Rats were randomly assigned to three groups:group Ⅰ:control,group Ⅱ: experimental colitis,group Ⅲ:colitis plus melatonin treatment.On d 11 after colitis,plasma tumor necrosis factor-α,portal blood endotoxin levels,colon tissue myeloperoxidase and caspase-3 activity were measured.Bacterial translocation was quantified by blood,lymph node,liver and spleen culture.RESULTS:We observed a significantly reduced incidence of bacterial translocation to the liver,spleen,mesenteric lymph nodes,portal and systemic blood in animals treated with melatonin.Treatment with melatonin significantly decreased the caspase-3 activity in colonic tissues compared to that in trinitrobenzene sulphonic acid-treated rats (16.11 ± 2.46 vs 32.97 ± 3.91,P < 0.01).CONCLUSION:Melatonin has a protective effect on bacterial translocation and apoptosis.

  16. Interfering with bacterial gossip

    DEFF Research Database (Denmark)

    Bjarnsholt, Thomas; Tolker-Nielsen, Tim; Givskov, Michael

    2011-01-01

    defense. Antibiotics exhibit a rather limited effect on biofilms. Furthermore, antibiotics have an ‘inherent obsolescence’ because they select for development of resistance. Bacterial infections with origin in bacterial biofilms have become a serious threat in developed countries. Pseudomonas aeruginosa...... that appropriately target bacteria in their relevant habitat with the aim of mitigating their destructive impact on patients. In this review we describe molecular mechanisms involved in “bacterial gossip” (more scientifically referred to as quorum sensing (QS) and c-di-GMP signaling), virulence, biofilm formation...

  17. 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.

  18. Multimodal analysis of ocular inflammation using the endotoxin-induced uveitis mouse model.

    Science.gov (United States)

    Chu, Colin J; Gardner, Peter J; Copland, David A; Liyanage, Sidath E; Gonzalez-Cordero, Anai; Kleine Holthaus, Sophia-Martha; Luhmann, Ulrich F O; Smith, Alexander J; Ali, Robin R; Dick, Andrew D

    2016-04-01

    Endotoxin-induced uveitis (EIU) in rodents is a model of acute Toll-like receptor 4 (TLR4)-mediated organ inflammation, and has been used to model human anterior uveitis, examine leukocyte trafficking and test novel anti-inflammatory therapeutics. Wider adoption has been limited by the requirement for manual, non-specific, cell-count scoring of histological sections from each eye as a measure of disease severity. Here, we describe a comprehensive and efficient technique that uses ocular dissection and multimodal tissue analysis. This allows matched disease scoring by multicolour flow cytometric analysis of the inflammatory infiltrate, protein analysis on ocular supernatants and qPCR on remnant tissues of the same eye. Dynamic changes in cell populations could be identified and mapped to chemokine and cytokine changes over the course of the model. To validate the technique, dose-responsive suppression of leukocyte infiltration by recombinant interleukin-10 was demonstrated, as well as selective suppression of the monocyte (CD11b+Ly6C+) infiltrate, in mice deficient for eitherCcl2orCcr2 Optical coherence tomography (OCT) was used for the first time in this model to allowin vivoimaging of infiltrating vitreous cells, and correlated with CD11b+Ly6G+ counts to provide another unique measure of cell populations in the ocular tissue. Multimodal tissue analysis of EIU is proposed as a new standard to improve and broaden the application of this model. PMID:26794131

  19. Multimodal analysis of ocular inflammation using the endotoxin-induced uveitis mouse model

    Directory of Open Access Journals (Sweden)

    Colin J. Chu

    2016-04-01

    Full Text Available Endotoxin-induced uveitis (EIU in rodents is a model of acute Toll-like receptor 4 (TLR4-mediated organ inflammation, and has been used to model human anterior uveitis, examine leukocyte trafficking and test novel anti-inflammatory therapeutics. Wider adoption has been limited by the requirement for manual, non-specific, cell-count scoring of histological sections from each eye as a measure of disease severity. Here, we describe a comprehensive and efficient technique that uses ocular dissection and multimodal tissue analysis. This allows matched disease scoring by multicolour flow cytometric analysis of the inflammatory infiltrate, protein analysis on ocular supernatants and qPCR on remnant tissues of the same eye. Dynamic changes in cell populations could be identified and mapped to chemokine and cytokine changes over the course of the model. To validate the technique, dose-responsive suppression of leukocyte infiltration by recombinant interleukin-10 was demonstrated, as well as selective suppression of the monocyte (CD11b+Ly6C+ infiltrate, in mice deficient for either Ccl2 or Ccr2. Optical coherence tomography (OCT was used for the first time in this model to allow in vivo imaging of infiltrating vitreous cells, and correlated with CD11b+Ly6G+ counts to provide another unique measure of cell populations in the ocular tissue. Multimodal tissue analysis of EIU is proposed as a new standard to improve and broaden the application of this model.

  20. New Cyt-like δ-endotoxins from Dickeya dadantii: structure and aphicidal activity.

    Science.gov (United States)

    Loth, Karine; Costechareyre, Denis; Effantin, Géraldine; Rahbé, Yvan; Condemine, Guy; Landon, Céline; da Silva, Pedro

    2015-01-01

    In the track of new biopesticides, four genes namely cytA, cytB, cytC and cytD encoding proteins homologous to Bacillus thuringiensis (Bt) Cyt toxins have been identified in the plant pathogenic bacteria Dickeya dadantii genome. Here we show that three Cyt-like δ-endotoxins from D. dadantii (CytA, CytB and CytC) are toxic to the pathogen of the pea aphid Acyrthosiphon pisum in terms of both mortality and growth rate. The phylogenetic analysis of the comprehensive set of Cyt toxins available in genomic databases shows that the whole family is of limited taxonomic occurrence, though in quite diverse microbial taxa. From a structure-function perspective the 3D structure of CytC and its backbone dynamics in solution have been determined by NMR. CytC adopts a cytolysin fold, structurally classified as a Cyt2-like protein. Moreover, the identification of a putative lipid binding pocket in CytC structure, which has been probably maintained in most members of the Cyt-toxin family, could support the importance of this lipid binding cavity for the mechanism of action of the whole family. This integrative approach provided significant insights into the evolutionary and functional history of D. dadantii Cyt toxins, which appears to be interesting leads for biopesticides.

  1. Endotoxin-induced monocytic microparticles have contrasting effects on endothelial inflammatory responses.

    Directory of Open Access Journals (Sweden)

    Beryl Wen

    Full Text Available Septic shock is a severe disease state characterised by the body's life threatening response to infection. Complex interactions between endothelial cells and circulating monocytes are responsible for microvasculature dysfunction contributing to the pathogenesis of this syndrome. Here, we intended to determine whether microparticles derived from activated monocytes contribute towards inflammatory processes and notably vascular permeability. We found that endotoxin stimulation of human monocytes enhances the release of microparticles of varying phenotypes and mRNA contents. Elevated numbers of LPS-induced monocytic microparticles (mMP expressed CD54 and contained higher levels of transcripts for pro-inflammatory cytokines such as TNF, IL-6 and IL-8. Using a prothrombin time assay, a greater reduction in plasma coagulation time was observed with LPS-induced mMP than with non-stimulated mMP. Co-incubation of mMP with the human brain endothelial cell line hCMEC/D3 triggered their time-dependent uptake and significantly enhanced endothelial microparticle release. Unexpectedly, mMP also modified signalling pathways by diminishing pSrc (tyr416 expression and promoted endothelial monolayer tightness, as demonstrated by endothelial impedance and permeability assays. Altogether, these data strongly suggest that LPS-induced mMP have contrasting effects on the intercellular communication network and display a dual potential: enhanced pro-inflammatory and procoagulant properties, together with protective function of the endothelium.

  2. 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.

  3. Adrenoceptor hyporeactivity is responsible for Escherichia coli endotoxin-induced acute vascular dysfunction in humans.

    Science.gov (United States)

    Pleiner, Johannes; Heere-Ress, Elisabeth; Langenberger, Herbert; Sieder, Anna E; Bayerle-Eder, Michaela; Mittermayer, Fritz; Fuchsjäger-Mayrl, Gabriele; Böhm, Johannes; Jansen, Burkhard; Wolzt, Michael

    2002-01-01

    Impaired response to catecholamines contributes to the altered hemodynamics in sepsis, which has been attributed to excessive NO formation. We have studied the systemic hemodynamic and local forearm responses and inducible NO synthase (iNOS) expression during experimental endotoxemia in humans. Escherichia coli endotoxin (lipopolysaccharide [LPS]) was administered at doses of 1 or 2 ng/kg to healthy volunteers. In 10 subjects, the systemic pressor effect of phenylephrine was assessed before and after the administration of LPS. In 9 further subjects, forearm blood flow responses to intra-arterial noradrenaline, acetylcholine, glyceryl trinitrate, and N(G)-monomethyl-L-arginine (L-NMMA) were studied at baseline and after LPS administration. Peripheral blood was collected and analyzed for iNOS mRNA and protein. Four hours after LPS, the response of systolic blood pressure (P<0.0005) and heart rate (P<0.05) to phenylephrine was significantly reduced. In the forearm, noradrenaline-induced vasoconstriction was also reduced by approximately 50% (P<0.01), but L-NMMA responsiveness was unchanged. iNOS mRNA or protein was not increased. Marked vascular adrenoceptor hyporeactivity is detectable in the absence of increased NO activity or iNOS expression in endotoxemia, arguing against major involvement of vascular iNOS activity in the acute systemic vasodilation to LPS.

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

    Directory of Open Access Journals (Sweden)

    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.

  5. Heterogeneity of epigenetic changes at ischemia/reperfusion- and endotoxin-induced acute kidney injury genes.

    Science.gov (United States)

    Mar, Daniel; Gharib, Sina A; Zager, Richard A; Johnson, Ali; Denisenko, Oleg; Bomsztyk, Karol

    2015-10-01

    Aberrant gene expression is a molecular hallmark of acute kidney injury (AKI). As epigenetic processes control gene expression in a cell- and environment-defined manner, understanding the epigenetic pathways that regulate genes altered by AKI may open vital new insights into the complexities of disease pathogenesis and identify possible therapeutic targets. Here we used matrix chromatin immunoprecipitation and integrative analysis to study 20 key permissive and repressive epigenetic histone marks at transcriptionally induced Tnf, Ngal, Kim-1, and Icam-1 genes in mouse models of AKI; unilateral renal ischemia/reperfusion, lipopolysaccharide (LPS), and their synergistically injurious combination. Results revealed unexpected heterogeneity of transcriptional and epigenetic responses. Tnf and Ngal were transcriptionally upregulated in response to both treatments individually, and to combination treatment. Kim-1 was induced by ischemia/reperfusion and Icam-1 by LPS only. Epigenetic alterations at these genes exhibited distinct time-dependent changes that shared some similarities, such as reduction in repressive histone modifications, and also had major ischemia/reperfusion versus endotoxin differences. Thus, diversity of changes at AKI genes in response to different insults indicates involvement of several epigenetic pathways. This could be exploited pharmacologically through rational-drug design to alter the course and improve clinical outcomes of this syndrome. PMID:26061546

  6. Multimodal analysis of ocular inflammation using the endotoxin-induced uveitis mouse model

    Science.gov (United States)

    Chu, Colin J.; Gardner, Peter J.; Copland, David A.; Liyanage, Sidath E.; Gonzalez-Cordero, Anai; kleine Holthaus, Sophia-Martha; Luhmann, Ulrich F. O.; Smith, Alexander J.; Ali, Robin R.; Dick, Andrew D.

    2016-01-01

    ABSTRACT Endotoxin-induced uveitis (EIU) in rodents is a model of acute Toll-like receptor 4 (TLR4)-mediated organ inflammation, and has been used to model human anterior uveitis, examine leukocyte trafficking and test novel anti-inflammatory therapeutics. Wider adoption has been limited by the requirement for manual, non-specific, cell-count scoring of histological sections from each eye as a measure of disease severity. Here, we describe a comprehensive and efficient technique that uses ocular dissection and multimodal tissue analysis. This allows matched disease scoring by multicolour flow cytometric analysis of the inflammatory infiltrate, protein analysis on ocular supernatants and qPCR on remnant tissues of the same eye. Dynamic changes in cell populations could be identified and mapped to chemokine and cytokine changes over the course of the model. To validate the technique, dose-responsive suppression of leukocyte infiltration by recombinant interleukin-10 was demonstrated, as well as selective suppression of the monocyte (CD11b+Ly6C+) infiltrate, in mice deficient for either Ccl2 or Ccr2. Optical coherence tomography (OCT) was used for the first time in this model to allow in vivo imaging of infiltrating vitreous cells, and correlated with CD11b+Ly6G+ counts to provide another unique measure of cell populations in the ocular tissue. Multimodal tissue analysis of EIU is proposed as a new standard to improve and broaden the application of this model. PMID:26794131

  7. Analysis of unmodified endotoxin preparations by 252Cf plasma desorption mass spectrometry. Determination of molecular masses of the constituent native lipopolysaccharides.

    Science.gov (United States)

    Caroff, M; Deprun, C; Karibian, D; Szabó, L

    1991-10-01

    Nine unmodified endotoxin preparations constituted of Re-, Rd-, and Rc-type lipopolysaccharides (2 to 5 glycoses), representing four species of enterobacteria were analyzed by 252Cf plasma desorption mass spectrometry. The constituent lipopolysaccharides were characterized by the ion pair: (M-H)- and its corresponding lipid fragment ion. The lipid fragment ion is produced by cleavage of the glycosidic bond of the 3-deoxy-D-manno-oct-2-ulosonic acid unit that substitutes O-6' of the glucosamin beta 1'-6glucosamine ("lipid A backbone") disaccharide of the lipid A moiety. These lipid fragment ions were identical to the (M-H)- ions seen in the spectra of homologous isolated lipid A preparations that were obtained by hydrolysis (pH 4.5, 100 degrees C) promoted by sodium dodecyl sulfate. Since the molecular components present in the endotoxin preparations analyzed are known, the ion pair (M-H)(-)-lipid fragment ion defines the molecular compositions of each individual lipopolysaccharide. Heterogeneity of the R-type endotoxin preparations analyzed was due almost exclusively to differing lipid A moieties. In three Salmonella minnesota 595 Re endotoxin preparations 10 different lipopolysaccharides were identified, only two of which were common to all three preparations. Of the nine lipopolysaccharides identified in two S. minnesota R7 endotoxin preparations, only two were present in both.

  8. Effects of garlic oil and two of its major organosulfur compounds, diallyl disulfide and diallyl trisulfide, on intestinal damage in rats injected with endotoxin

    International Nuclear Information System (INIS)

    Garlic and its active components are known to possess antioxidant and antiinflammatory effects. The present study investigated the effects of garlic oil and its organosulfur compounds on endotoxin-induced intestinal mucosal damage. Wistar rats received by gavage 50 or 200 mg/kg body weight garlic oil (GO), 0.5 mmol/kg body weight diallyl disulfide or diallyl trisulfide, or the vehicle (corn oil; 2 ml/kg body weight) every other day for 2 weeks before being injected with endotoxin (i.p., 5 mg/kg body weight). Control rats were administered with corn oil and were injected with sterile saline. Samples for the measurement of proinflammatory cytokines were collected 3 h after injection, and all other samples were collected 18 h after injection. The low dose of GO suppressed endotoxin-induced inducible nitric oxide synthase (iNOS) activity, ulceration, and apoptosis in the intestinal mucosa (P < 0.05). The high dose of GO significantly lowered the peripheral level of nitrate/nitrite and endotoxin-induced iNOS activity in the intestinal mucosa (P < 0.05) but worsened intestinal mucosal damage accompanied by elevated peripheral proinflammatory cytokines. Diallyl trisulfide but not diallyl disulfide showed similar toxic effect as that of high-dose GO. These results suggest the preventive effect and possible toxicity of garlic oil and its organosulfur compounds in endotoxin-induced systemic inflammation and intestinal damage

  9. Bacterial Wound Culture

    Science.gov (United States)

    ... Home Visit Global Sites Search Help? Bacterial Wound Culture Share this page: Was this page helpful? Also known as: Aerobic Wound Culture; Anaerobic Wound Culture Formal name: Culture, wound Related ...

  10. Bacterial surface adaptation

    Science.gov (United States)

    Utada, Andrew

    2014-03-01

    Biofilms are structured multi-cellular communities that are fundamental to the biology and ecology of bacteria. Parasitic bacterial biofilms can cause lethal infections and biofouling, but commensal bacterial biofilms, such as those found in the gut, can break down otherwise indigestible plant polysaccharides and allow us to enjoy vegetables. The first step in biofilm formation, adaptation to life on a surface, requires a working knowledge of low Reynolds number fluid physics, and the coordination of biochemical signaling, polysaccharide production, and molecular motility motors. These crucial early stages of biofilm formation are at present poorly understood. By adapting methods from soft matter physics, we dissect bacterial social behavior at the single cell level for several prototypical bacterial species, including Pseudomonas aeruginosa and Vibrio cholerae.

  11. Bacterial intermediate filaments

    DEFF Research Database (Denmark)

    Charbon, Godefroid; Cabeen, M.; Jacobs-Wagner, C.

    2009-01-01

    Crescentin, which is the founding member of a rapidly growing family of bacterial cytoskeletal proteins, was previously proposed to resemble eukaryotic intermediate filament (IF) proteins based on structural prediction and in vitro polymerization properties. Here, we demonstrate that crescentin...

  12. Bacterial Meningitis in Infants

    Directory of Open Access Journals (Sweden)

    J Gordon Millichap

    2004-04-01

    Full Text Available A retrospective study of 80 infantile patients (ages 30-365 days; 47 male, 33 female with culture-proven bacterial meningitis seen over a 16 year period (1986-2001 is reported from Taiwan.

  13. Featherweight Challenge

    Science.gov (United States)

    Love, Tyler S.; Ryan, Larry

    2012-01-01

    As science, technology education, and engineering programs suffer budget cuts, educators continue to seek cost-effective activities that engage students and reinforce standards. The featherweight challenge is a hands-on activity that challenges students to continually refine their design while not breaking the budget. This activity uses one of the…

  14. Immunization by a bacterial aerosol.

    Science.gov (United States)

    Garcia-Contreras, Lucila; Wong, Yun-Ling; Muttil, Pavan; Padilla, Danielle; Sadoff, Jerry; Derousse, Jessica; Germishuizen, Willem Andreas; Goonesekera, Sunali; Elbert, Katharina; Bloom, Barry R; Miller, Rich; Fourie, P Bernard; Hickey, Anthony; Edwards, David

    2008-03-25

    By manufacturing a single-particle system in two particulate forms (i.e., micrometer size and nanometer size), we have designed a bacterial vaccine form that exhibits improved efficacy of immunization. Microstructural properties are adapted to alter dispersive and aerosol properties independently. Dried "nanomicroparticle" vaccines possess two axes of nanoscale dimensions and a third axis of micrometer dimension; the last one permits effective micrometer-like physical dispersion, and the former provides alignment of the principal nanodimension particle axes with the direction of airflow. Particles formed with this combination of nano- and micrometer-scale dimensions possess a greater ability to aerosolize than particles of standard spherical isotropic shape and of similar geometric diameter. Here, we demonstrate effective application of this biomaterial by using the live attenuated tuberculosis vaccine bacille Calmette-Guérin (BCG). Prepared as a spray-dried nanomicroparticle aerosol, BCG vaccine exhibited high-efficiency delivery and peripheral lung targeting capacity from a low-cost and technically simple delivery system. Aerosol delivery of the BCG nanomicroparticle to normal guinea pigs subsequently challenged with virulent Mycobacterium tuberculosis significantly reduced bacterial burden and lung pathology both relative to untreated animals and to control animals immunized with the standard parenteral BCG. PMID:18344320

  15. 降钙素原联合内毒素鲎试验在儿童社区获得性肺炎中的临床应用%The clinical application of procalcitonin and endotoxin limulus in community - acquired pneumonia of children

    Institute of Scientific and Technical Information of China (English)

    王萍; 王华彬; 罗开源

    2016-01-01

    Objective:To evaluate the clinical diagnostic value of procalcitonin(PCT)combined with endotoxin Limulus test in children community - acquired pneumonia. Methods:From January 2014 to December 2015,admissions 120 cases of community - acquired pneumonia in children,according to etiology findings will be divided into groups of bacteria and non -bacterial group,the group of children with bacterial subdivided into Gram - negative bacteria infection group(A group)and non - Gram - negative bacteria infection group(group B),all children with procalcitonin,quantitative determination of endo-toxin limulus test,observe the test results. Results:According to test results can be seen in children with positive rates of bacte-rial infection procalcitonin group was higher than that of children of non - bacterial group,the comparison of the data between the two groups of children was significantly(P ﹤ 0. 05),with statistical significance. Endotoxin by the Limulus test,found that children with serum toxin A group changed significantly in the serum group B toxin no significant change,data comparison be-tween the two groups of children was significantly(P ﹤ 0. 05),with statistical significance. Conclusion:Serum procalcitonin for early diagnosis of children with community - acquired pneumonia have greater clinical value,quantitative LAL endotoxin assay can determine the type of bacteria,to prevent the wrong use of antibiotics,due to the direction of reducing medication errors ef-fective treatment for clinical medicine has important reference value.%目的:探讨降钙素原(PCT)联合内毒素鲎试验在儿童社区获得性肺炎临床诊断中的应用价值。方法:选取2014年1月—2015年12月接诊的120例社区获得性肺炎患儿,根据病原学检查结果将其分成细菌组与非细菌组,将细菌性组患儿再分为革兰阴性菌感染组(A 组)和非革兰阴性菌感染组(B 组),将所有患儿进行降钙素原、内毒素鲎定量

  16. Expression of cyclin D{sub 1} during endotoxin-induced aleveolar type II cell hyperplasia in rat lung and the detection of apoptotic cells during the remodeling process

    Energy Technology Data Exchange (ETDEWEB)

    Tesfaigzi, J.; Wood, M.B.; Johnson, N.F.

    1995-12-01

    Our studies have shown that endotoxin intratracheally instilled into the rat lung induces proliferation of alveolar type II cells. In that study, the alveolar type II cells. In that study, the alveolar type II cell hyperplasia occurred 2 d after instillation of endotoxin and persisted for a further 2 d. After hyperplasia, the lung remodeled and returned to a normal state within 24-48 h. Understanding the mechanisms involved in the remodeling process of this transient hyperplasia may be useful to identify molecular changes that are altered in neoplasia. The purpose of the present study was to corroborate induction of epithelial cell hyperplasia by endotoxin and to delineate mechanisms involved in tissue remodeling after endotoxin-induced alveolar type II cell hyperplasia. In conclusion, immonostaining with cyclin D1 and cytokeratin shows that endotoxin induced epithelial cell proliferation and resulted in hyperplasia in the lung which persisted through 4 d post-instillation.

  17. Challenging Identities

    DEFF Research Database (Denmark)

    Keydar, Ceglar; Marin, Manuela; Janik, Allan;

    , cultural, and political practices. Notions of national identity and national politics are challenged by European integration, as well as by increasing demographic heterogeneity due to migration, and migrants experience conflicts of identification stemming from clashes between cultural heritage...

  18. 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. PMID:12785015

  19. The bacterial lipocalins.

    Science.gov (United States)

    Bishop, R E

    2000-10-18

    The lipocalins were once regarded as a eukaryotic protein family, but new members have been recently discovered in bacteria. The first bacterial lipocalin (Blc) was identified in Escherichia coli as an outer membrane lipoprotein expressed under conditions of environmental stress. Blc is distinguished from most lipocalins by the absence of intramolecular disulfide bonds, but the presence of a membrane anchor is shared with two of its closest homologues, apolipoprotein D and lazarillo. Several common features of the membrane-anchored lipocalins suggest that each may play an important role in membrane biogenesis and repair. Additionally, Blc proteins are implicated in the dissemination of antibiotic resistance genes and in the activation of immunity. Recent genome sequencing efforts reveal the existence of at least 20 bacterial lipocalins. The lipocalins appear to have originated in Gram-negative bacteria and were probably transferred horizontally to eukaryotes from the endosymbiotic alpha-proteobacterial ancestor of the mitochondrion. The genome sequences also reveal that some bacterial lipocalins exhibit disulfide bonds and alternative modes of subcellular localization, which include targeting to the periplasmic space, the cytoplasmic membrane, and the cytosol. The relationships between bacterial lipocalin structure and function further illuminate the common biochemistry of bacterial and eukaryotic cells.

  20. Exposition aux endotoxines aéroportées et allergènes dans les animaleries et risques pour la santé

    OpenAIRE

    SCHWERY M.

    2014-01-01

    L'exposition aux endotoxines peut présenter des effets bénéfiques sur la santé. En effet, plusieurs études démontrent leur importance dans la théorie hygiéniste ainsi que leur effet négatif sur la croissance de certaines tumeurs. Toutefois, dans divers secteurs professionnels, l'environnement de travail favorise la présence d'endotoxines aéroportées et peut exposer le personnel à des risques sanitaires. C'est le cas...