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Sample records for bacterial endotoxin pyrogen

  1. 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

  2. An investigation of the possibility to replace the rabbit pyrogen test by an in vitro test

    NARCIS (Netherlands)

    Gommer AM; Donders LAM; LGM

    1998-01-01

    The classical test for the detection of pyrogenic contaminations in pharmaceuticals is the rabbit pyrogen test. The most frequently occuring and most important pyrogenic contamination in biological and biotechnological pharmaceutical preparations are bacterial endotoxins, originating from the cellul

  3. Applicability of bacterial endotoxins test to various blood products by the use of endotoxin-specific lysates.

    Science.gov (United States)

    Ochiai, Masaki; Yamamoto, Akihiko; Naito, Seishiro; Maeyama, Jun-Ichi; Masumi, Atsuko; Hamaguchi, Isao; Horiuchi, Yoshinobu; Yamaguchi, Kazunari

    2010-11-01

    Endotoxin contamination is a serious threat to the safety of parenteral drugs, and the rabbit pyrogen test has played a crucial role in controlling this contamination. Although the highly sensitive endotoxin test has replaced the pyrogen test for various pharmaceuticals, the pyrogen test is still implemented as the control test for most blood products in Japan. We examined the applicability of the endotoxin test to blood products for reliable detection and quantification of endotoxin. Nineteen types of blood products were tested for interfering factors based on spike/recovery of endotoxin by using 2 types of endotoxin-specific lysate reagents for photometric techniques. Interfering effects on the endotoxin test by the products could be eliminated by diluting from 1/2 to 1/16, with the exception of antithrombin III. However, conventional lysate reagents that also react with non-pyrogenic substances, such as (1-3)-β-D-glucan, produced results that were not relevant to endotoxin content or pyrogenicity. Our results showed that the endotoxin test would be applicable to most blood products if used with appropriate endotoxin-specific lysate reagents.

  4. 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.

  5. [Role of the adenyl cyclase system in achieving the immunogenesis-stimulating action of bacterial lipopolysaccharides--pyrogenal and endogenous serum pyrogen].

    Science.gov (United States)

    Dzheksenbaev, O Sh; Selezneva, V P; Loginova, V T

    1976-05-01

    Experiments were conducted on rabbits immunized intraperitoneally with corpuscular typhoid vaccine; the number of antibody-forming cells in the spleen proved to increase after tha administration of bacterial lipopolysaccharide (LPS)--pyropeneal, and endogenous serum pyrogen (EPS) together with theopylline. The data obtained indicated that the adenylcyclase system played a certain role in the mechanism of the stimulating action of LPS and EPS.

  6. 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.

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

    Science.gov (United States)

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

    2014-04-01

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

  8. A selective sorbent for removing bacterial endotoxins from blood

    Science.gov (United States)

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

    2016-12-01

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

  9. 丹红注射液中细菌内毒素的检测%Discussion on the bacterial endotoxin test of Danhong Injection

    Institute of Scientific and Technical Information of China (English)

    张秀花

    2011-01-01

    Objective: To explore the feasibility of the bacterial endotoxin test for Danhong Injection. Methods: The experiment was based on the bacterial endotoxin test method stated in Chinese Pharmacoperia 2010 Edition (Vol II), which helped to identify the effective dilution of Danhong Injection and the limits of bacteria endotoxin. Results: When the Danhong Injection was diluted 10 times, it had no interference effects on the TAL. The limit of bacterial endotoxin was 5.0 ELJ/ ml. Conclusion: The bacterial endotoxin test can be used to replace the pyrogen test.%目的:探讨丹红注射液细菌内毒素检查方法的可行性.方法:依据2010年版二部附录细菌内毒素检查法,确定丹红注射液的有效稀释浓度和细菌内毒素限值.结果:丹红注射液经稀释10倍后对鲎试剂无干扰作用,细菌内毒素的限值为5.0 EU/ml.结论:可用细菌内毒素检查取代热原检查.

  10. 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.

  11. 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.

  12. 家兔法与内毒素法检查生物材料热原的适用性研究%Applicability Research for the Bacteria Endotoxin Test Method and Rabbit test Method to Detect Pyrogen

    Institute of Scientific and Technical Information of China (English)

    邹文; 袁暾; 蔡永福; 梁洁

    2011-01-01

    To evaluate drugs and biomaterials have great difference. Bacteria endotoxin test was widely applied to detect drug pyrogen. However, if it is Appropriate to detect pyrogen by bacteria endotoxin test for some materials which has complex components, this must to be definituded. At the base of pharmacopoeia (2005), pyrogen was detected for each tissue engineering bioamterial to use rabbit method and the bacteria endotoxin method individually. The result was indicated to be negative for the bacteria endotoxin method and positive for the rabbit method. Hie pyrogen test result was not accord with the two methodes. The result indicated the pyrogen for the product of some biomaterialfc was complicated. The rabbit test method is more safe and reliable than the bacteria endotoxin test method.%药品与生物材料在生物安全性评价方法上有着很大的区别.临床上广泛运用内毒素法检查药品热原,然而,运用细菌内毒素法进行部分组成成分较为复杂的生物材料的热原试验是否适当有待明确.本研究在2005版药典的基础上,分别运用内毒素法和家兔法对两种组织工程支架材料进行热原试验的比较研究,实验结果表明运用内毒素法得到的试验结果为阴性,运用家兔法得到的试验结果为阳性.这两种方法分别测定每种材料所得到的热原试验结果不相符合,表明对组成成分复杂的生物材料,含热原的因素较为复杂,用家兔法进行试验检测热原可能更加灵敏.

  13. 注射用阿魏酸钠细菌内毒素检查法的建立%Studies on Establishment of Bacterial Endotoxin Test of Sodium Ferulate for Injection

    Institute of Scientific and Technical Information of China (English)

    王相忍

    2014-01-01

    目的:建立注射用阿魏酸钠细菌内毒素检查法。方法采用《中国药典》2010年版二部附录细菌内毒素检查法。结果注射用阿魏酸钠稀释至1.25mg· mL -1时,不干扰细菌内毒素试验,其细菌内毒素限值可订为0·4EU· mg -1。结论使用细菌内毒素检查法检查注射用阿魏酸钠中的细菌内毒素是可行的,可用细菌内毒素检查法代替家兔热原检查法。%OBJECTIVE To establish a method for the bacterial endotoxin test of Sodium Ferulate for Injec-tion.METHODS The method of the bacterial endotoxin test approved by the Chinese Pharmacopoeia ( edition 2010 ) was adopted.RESULTS Sodium Ferulate for Injection was diluted to 1.25 mg· mL-1 ,does not interfere with the bacterial endotoxin test ,the bacterial endotoxin limit can be set for 0.4EU· mg-1.CONCLUSION The meth-od is feasible to detect bacterial endotoxin in Sodium Ferulate for Injection , which can replace the rabbit pyrogen test.

  14. 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).

  15. 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).

  16. 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)

  17. 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.

  18. 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)

  19. Detection of bacterial endotoxin in drinking tap and bottled water in Kuwait.

    Science.gov (United States)

    Abdulraheem, Abdulkareem; Mustafa, Seham; Al-Saffar, Nabeel; Shahjahan, Muhammed

    2012-12-01

    This study was carried out to measure and compare the concentration of bacterial endotoxin in a variety of samples from drinking tap and bottled water available in Kuwait by using the Limulus Amoebocyte lysate test. A total of 29 samples were tested. Samples were collected from a variety of locations throughout the six governorates of Kuwait and 23 brands of local and imported bottled water samples were collected from the local market. The concentration of bacterial endotoxin was measured by using the standard Limulus Amoebocyte lysate test, gel clot method. This study showed that measured endotoxin concentrations in tap drinking water varied from 2.4 to 33.8 EU/ml with the average endotoxin concentration of 14.2 EU/ml. While the results of endotoxin concentrations in the bottled water were bottled water is 13.5 % of the average concentration of endotoxin in tap drinking water. This experimental investigation has proved that drinking bottled water has less endotoxin as compared to tap water in Kuwait. It is also demonstrated that the endotoxin concentration did not exceed the acceptable level in drinking tap water.

  20. 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.

  1. 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.

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

    Science.gov (United States)

    Shi, Xingmin; Li, Yaxi; Zhang, Guanjun; Ma, Yue; Shao, Xianjun

    2011-12-01

    The low-temperature plasma (LTP) generated by dielectric barrier discharge (DBD) was used to sterilize the E.coli endotoxin, which is usually difficult to kill by traditional methods. Three different concentrations of bacterial endotoxin (1 EU/mL, 0.5 EU/mL and 0.25 EU/mL) were treated by LTP for different time (20 s, 40 s and 60 s). Tachypleus amebocyte lysate (TAL) method was employed to detect the concentration variation of bacterial endotoxin before and after the plasma treatment, and endotoxic shock mice model was used to evaluate the inactivation effects of LTP on endotoxin for further study. Experimental results demonstrated that, DBD plasma can inactivate the bacterial endotoxin quickly and effectively, and when the LTP treatment time was increased, the concentrations of bacterial endotoxin decreased gradually (after 60 s plasma treatment, its inactivation effect was beyond the Chinese pharmacopoeia standard), and the average survival time of mice gradually extended. The possible inactivation mechanisms are proposed to be related to reactive oxygen species (ROSs).

  3. Endotoxin Detection in Pharmaceuticals and Medical Devices with Kinetic-QCL, a Kinetic-Quantitative Chromogenic Limulus Amebocyte Lysate Assay.

    Science.gov (United States)

    Berzofsky, Ronald N.

    1995-01-01

    The observation that endotoxin caused gelation in extracts of Limulus amebocytes has been expanded to the development of an in vitro kinetic, quantitative chromogenic LAL assay (Kinetic-QCL) for the detection of endotoxin in aqueous fluids. Within the last 15 years, the use of Limulus amebocyte lysate to detect and control the presence of pyrogenic substances in pharmaceuticals and medical devices has gained wide international acceptance. Both the United States and European Pharmacopoeias contain descriptions of and requirements for the LAL Bacterial Endotoxin Test. Both pharmacopoeias have begun to remove the rabbit pyrogen test requirement in a majority of drug monographs and have substituted endotoxin limits to be determined by LAL. The use of LAL has proved invaluable in controlling the level of endotoxin in finished product. The endotoxin contribution of raw materials and packaging material can be monitored as well. In-process testing at critical production steps can identify additional sources of endotoxin contamination, and depyrogenation processes can be validated by quantitating the degradation of endotoxin challenges. The speed, reproducibility, sensitivity, and economics of the Kinetic-QCL assay, in conjunction with the ppropriate equipment and software, over both the in vivo rabbit pyrogen test and the more traditional LAL gel-clot assay allow a more in-depth approach to the control of endotoxin in pharmaceuticals and medical devices.

  4. Dialysate bacterial endotoxin as a prognostic indicator of peritoneal dialysis related peritonitis.

    Science.gov (United States)

    Szeto, Cheuk-Chun; Lai, Ka-Bik; Chow, Kai-Ming; Kwan, Bonnie Ching-Ha; Law, Man-Ching; Pang, Wing-Fai; Ma, Terry King-Wing; Leung, Chi-Bon; Li, Philip Kam-Tao

    2016-12-01

    Peritonitis is the major complication of peritoneal dialysis (PD). The aim of our present study is to explore the prognostic value of endotoxin level in PD effluent for the prediction of treatment failure in PD-related peritonitis. We studied 325 peritonitis episodes in 223 patients. PD effluent (PDE) was collected every 5 days for endotoxin level and leukocyte count. Patients were followed for relapsing or recurrent peritonitis. We found 20 episodes (6.2%) had primary treatment failure; 41 (12.6%) developed relapsing, 19 (5.8%) had recurrent, and 22 (6.8%) had repeat episodes. Endotoxin was detectable in the PDE of 19 episodes (24.4%) caused by Gram negative organisms, 4 episodes (6.8%) of mixed bacterial growth, and none of the culture negative episodes or those by Gram positive organisms. For episodes caused by Gram negative bacteria, a detectable endotoxin level in PDE on day 5 had a sensitivity and specificity of 66.7% and 83.3%, respectively, for predicting primary treatment failure. In contrast, PDE leukocyte count > 1000 per mm3 on day 5 had a sensitivity and specificity of 88.9% and 89.1%, respectively; the addition of PDE endotoxin assay did not improve the sensitivity or specificity. We conclude that detectable endotoxin in PDE 5 days after antibiotic therapy might predict primary treatment failure in peritonitis episodes caused by Gram negative organisms. However, the sensitivity and specificity of PDE endotoxin assay was inferior to PDE leukocyte count.

  5. Study on bacterial endotoxin test of iron sucrose injection%蔗糖铁注射液细菌内毒素检测法研究

    Institute of Scientific and Technical Information of China (English)

    任韡; 李欣

    2016-01-01

    目的:建立检查蔗糖铁注射液细菌内毒素的检测方法(凝胶法),以控制药物质量,减少临床热原反应的发生。方法:按《中国药典》2015年版(四部)收载的细菌内毒素检测方法及指导原则进行实验,系统观察蔗糖铁注射液对鲎试剂与细菌内毒素凝集反应的干扰,确定凝胶法检测细菌内毒素的不干扰浓度。结果:蔗糖铁注射液稀释20倍后对鲎试剂无干扰作用,对2个厂家的8批样品进行检查,结果均符合规定。结论:细菌内毒素检测方法(凝胶法)可用于蔗糖铁注射液的内毒素检查,结果准确、重现性好。%Objective:To set up the bacterial endotoxin determination method of iron sucrose injection (the gel-clot method) for the quality control of the drug and prevention of pyrogen reaction in clinical practice.Methods:The experiment was performed according to the Chinese Pharmacopoeia 2015 edition (4). A systematic study was carried out to investigate the interferences of iron sucrose injection with limulus reagent and agglutination reaction to bacterial endotoxin in order to detect the non-interference concentration of bacterial endotoxin.Results: There was no interference in the bacterial endotoxin test when iron sucrose injection was diluted 20-fold. The results of 8 batches of samples from 2 manufacturers complied with the requirements. Conclusion: The established method of endotoxin test was feasible, accurate and repeatable for iron sucrose injection.

  6. 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.

  7. 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

  8. 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。

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

    Lifescience Database Archive (English)

    Full Text Available y - PNG File (.png) SVG File (.svg) HTML File (.html) CSML File (.csml) Open .csml file with CIOPlayer Open .csm...n M. Ann N Y Acad Sci. 2007 Jan;1096:1-17. (.png) (.svg) (.html) (.csml) Show Targeting bacterial endotoxin:...l file with CIOPlayer - ※CIO Playerのご利用上の注意 Open .csml file with CIO Open .csml file with CIO - ※CIOのご利用上の注意 ...

  10. 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.

  11. Pyrogenic effect of respirable road dust particles

    Energy Technology Data Exchange (ETDEWEB)

    Jayawardena, Umesh; Tollemark, Linda; Tagesson, Christer; Leanderson, Per, E-mail: per.leanderson@lio.s [Occupational and Environmental Medicine, University Hospital, S-581 85 Linkoeping (Sweden)

    2009-02-01

    Because pyrogenic (fever-inducing) compounds on ambient particles may play an important role for particle toxicity, simple methods to measure pyrogens on particles are needed. Here we have used a modified in vitro pyrogen test (IPT) to study the release of interleukin 1beta (IL-1beta) in whole human blood exposed to respirable road-dust particles (RRDP). Road dusts were collected from the roadside at six different streets in three Swedish cities and particles with a diameter less than 10 mum (RRDP) were prepared by a water sedimentation procedure followed by lyophilisation. RRDP (200 mul of 1 - 10{sup 6} ng/ml) were mixed with 50 mul whole blood and incubated at 37 deg. C overnight before IL-1beta was analysed with chemiluminescence ELISA in 384-well plates. Endotoxin (lipopolysaccharide from Salmonella minnesota), zymosan B and Curdlan (P-1,3-glucan) were used as positive controls. All RRDP samples had a pyrogenic effect and the most active sample produced 1.6 times more IL-1beta than the least active. This formation was of the same magnitude as in samples with 10 ng LPS/ml and was larger than that evoked by zymosan B and Curdlan (by mass basis). The method was sensitive enough to determine formation of IL-1beta in mixtures with 10 ng RRDP/ml or 0.01 ng LPS/ml. The endotoxin inhibitor, polymyxin B (10 mug/ml), strongly reduced the RRDP-induced formation of IL-1beta at 1mug RRDP/ml (around 80 % inhibition), but had only marginal or no effects at higher RRDP-concentrations (10 and 100 mug /ml). In summary, all RRDP tested had a clear pyrogen effect in this in vitro model. Endotoxin on the particles but also other factors contributed to the pyrogenic effect. As opposed to the limulus amebocyte lysate (LAL) assay (which measures endotoxin alone), IPT measures a broad range of pyrogens that may be present on particulate matter. The IPT method thus affords a simple, sensitive and quantitative determination of the total pyrogenic potential of ambient particles.

  12. Research on Pyrogen Test by Replacing Domestic Rabbit with Whole Blood or Cell%用全血或细胞替代家兔进行热原检查的方法研究

    Institute of Scientific and Technical Information of China (English)

    李冠民; 黄清泉; 张云河; 贺争鸣

    2003-01-01

    The pyrogen test is an important index of quality-management relating to druggery injection and instillation and the current common methods are domestic-rabbit test and Bacterial Endotoxins Test. Since there are shortcomings and limitations of resources in these two methods , it's necessary to look for another new substitute. We have done the preliminary study of the feasibility of the pyrogen test with domestic rabbit's blood plasm, human's plasm and THP-1.The principle is as follows: after incubating the standard sample of bacterial endotoxins, glycogen anti-coagulation whole blood extracting from the healthy domestic rabbit, extralin anti-congealable whole blood from healthy volunteers,THP-1 cells together, measure the releasing amount of TNF and IL-6 from cell gene by the method of ELISA as a endogenesis index judging the pro-heat role of endotoxin.The results indicate: the blood-rabbit's whole blood, human's whole blood, or THP-lcelI, their releasing amount of TNF and IL-6 are closely connected with the amount of acting endotoxins within a certain range. Three methods have their own peculiarities and some feasibilities as a new substitute method for pyrogen test with domestic rabbit.

  13. Over-starvation aggravates intestinal injury and promotes bacterial and endotoxin translocation under high-altitude hypoxic environment

    Institute of Scientific and Technical Information of China (English)

    Qi-Quan Zhou; Ding-Zhou Yang; Yong-Jun Luo; Su-Zhi Li; Fu-Yu Liu; Guan-Song Wang

    2011-01-01

    AIM: To study whether over-starvation aggravates intestinal mucosal injury and promotes bacterial and endotoxin translocation in a high-altitude hypoxic environment. METHODS: Sprague-Dawley rats were exposed to hypobaric hypoxia at a simulated altitude of 7000 m for 72 h. Lanthanum nitrate was used as a tracer to detect intestinal injury. Epithelial apoptosis was observed with terminal deoxynucleotidyl transferase dUTP nick end labeling staining. Serum levels of diamino oxidase (DAO), malondialdehyde (MDA), glutamine (Gln), superoxide dismutase (SOD) and endotoxin were measured in intestinal mucosa. Bacterial translocation was detected in blood culture and intestinal homogenates. In addition, rats were given Gln intragastrically to observe its protective effect on intestinal injury. RESULTS: Apoptotic epithelial cells, exfoliated villi and inflammatory cells in intestine were increased with edema in the lamina propria accompanying effusion of red blood cells. Lanthanum particles were found in the intercellular space and intracellular compartment. Bacterial translocation to mesenteric lymph nodes (MLN) and spleen was evident. The serum endotoxin, DAO and MDA levels were significantly higher while the serum SOD, DAO and Gln levels were lower in intestine (P < 0.05). The bacterial translocation number was lower in the high altitude hypoxic group than in the high altitude starvation group (0.47 ± 0.83 vs 2.38 ± 1.45, P < 0.05). The bacterial translocation was found in each organ, especially in MLN and spleen but not in peripheral blood. The bacterial and endotoxin translocations were both markedly improved in rats after treatment with Gln. CONCLUSION: High-altitude hypoxia and starvation cause severe intestinal mucosal injury and increase bacterial and endotoxin translocation, which can be treated with Gln.

  14. 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...... experiment, fish were injected with bacterial endotoxin lipopolysaccharide (LPS) either at ZT3 (day) or at ZT15 (night) to evaluate the temporal sensitivity of humoral immunity to a pathogen-associated molecular pattern. The results demonstrated that responses to LPS were gated by the time of day. LPS...... significantly modulated serum ALP and ANTI activities but only when the endotoxin was administered at ZT3. Serum LYZ and PER were stimulated at both injection times but with differing response profiles. Modulated LYZ activity was persistent when injected at ZT3 but transient when LPS was applied at ZT15...

  15. Molecular requirements for MHC class II alpha-chain engagement and allelic discrimination by the bacterial superantigen streptococcal pyrogenic exotoxin C.

    Science.gov (United States)

    Kasper, Katherine J; Xi, Wang; Rahman, A K M Nur-Ur; Nooh, Mohammed M; Kotb, Malak; Sundberg, Eric J; Madrenas, Joaquín; McCormick, John K

    2008-09-01

    Superantigens (SAgs) are microbial toxins that bind to both TCR beta-chain variable domains (Vbetas) and MHC class II molecules, resulting in the activation of T cells in a Vbeta-specific manner. It is now well established that different isoforms of MHC II molecules can play a significant role in the immune response to bacterial SAgs. In this work, using directed mutational studies in conjunction with functional analyses, we provide a complete functional map of the low-affinity MHC II alpha-chain binding interface of the SAg streptococcal pyrogenic exotoxin C (SpeC) and identify a functional epitope in the beta-barrel domain that is required for the activation of T cells. Using cell lines that exclusively express individual MHC II isoforms, our studies provide a molecular basis for the selectivity of SpeC-MHC II recognition, and provide one mechanism by how SAgs are capable of distinguishing between different MHC II alleles.

  16. Methodological Studies on Plasma Endotoxin Level and Endotoxin Inactivation Capacity

    Institute of Scientific and Technical Information of China (English)

    姚国相; 杨乃发; 薛新波; 赵玉沛; 蒋朱明

    2004-01-01

    To establish stable methods for detecting plasma endotoxin level and endotoxin inactivation capacity in a normal population and general surgical patients and evaluate their perioperative changes, 50 healthy people and 50 patients receiving gastrointestinal operation were enrolled, their plasma endotoxin levels and plasma endotoxin inactivation capacity were assayed. Our results showed that plasma endotoxin levels were 0.044±0.009 EU/ml in the normal population and 0.044±0.023 EU/ml in the preoperative patients. Endotoxin level peaked 3 h after the operation (0.223±0.041 EU/ml), and then decreased rapidly on the first day after the operation (0.134±0.164EU/ml). Endotoxin inactivation capacity also had the same time course as endotoxin level. Systemic inflammatory response syndrome and infection induced another elevation in the time course. It is concluded that establishing the endotoxin standard curve by using pyrogenic free water is better than by using plasma. Plasma endotoxin inactivation capacity can be used as an indirect indicator of postoperative immune depression. Plasma endotoxin level and endotoxin inactivation capacity peaked shortly after operation, indicating surgical stress is closely related with the changes.

  17. 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.

  18. Bacterial colonization and endotoxin content of a new renal dialysis water system composed of acrylonitrile butadiene styrene.

    Science.gov (United States)

    du Moulin, G C; Coleman, E C; Hedley-Whyte, J

    1987-06-01

    We measured endotoxin and bacterial levels in tap water, in water purified by reverse osmosis, and in dialysate samples over a 4-month period in a new 10-bed renal dialysis unit. Water treated by reverse osmosis is conducted to the 10 stations through 111 m of piping composed of acrylonitrile butadiene styrene (ABS). All determinations were made prior to the opening of the unit and after the system was purged for 35 h with all bedside station taps open. Formaldehyde disinfection of the piping system was attempted with a recommended protocol after 11 weeks by feeding 2.5 liters of 37% formaldehyde (0.85%, vol/vol) into the delivery system. Prior to water purging, 24 ng of endotoxin per ml was detected. This level decreased to 2.0 ng of endotoxin after the purging. Levels of endotoxin remained below 1.0 ng of endotoxin per ml throughout the duration of the study. In contrast, the level of viable microorganisms recovered from the treated water was approximately 3.5 X 10(4) CFU/100 ml. Even after disinfection of the system, there was no significant decrease in culturable bacteria from the water even though endotoxin levels were lower. Species isolated from the renal dialysis system were predominately pseudomonads, whereas species isolated from the tap water were Bacillus and Flavobacterium species. ABS provides a surface suitable for long-term colonization and growth of bacteria. Currently recommended decontamination protocols are ineffective in removing potentially pathogenic bacteria from ABS pipes and thus constitute an increased risk to patients undergoing dialysis.

  19. Study on Establishment of Bacterial Endotoxin Test of Furosemide Injection%呋塞米注射液细菌内毒素检查法的建立

    Institute of Scientific and Technical Information of China (English)

    李逢春; 张广伟

    2012-01-01

    目的 建立呋塞来注射液的细菌内毒素检查法.方法 采用2005年版《中国药典(二部)》附录细菌内毒素检查法.结果 将呋塞米注射液稀释到质量浓度为0.8 g/L时,不干扰细菌内毒素试验,其细菌内毒素限值可订为1.2 EU/mg.结论 使用细菌内毒素检查法检查呋塞米注射液中的细菌内毒素是可行的.%Objective To establish a method for the bacterial endotoxin test of furosemide injection. Methods The method of the bacterial endotoxin test approved by the Chinese Pharmacopoeia (edition 2005) was adopted. Results If the solution of furosemide injection was diluted to 0.8 g/L,the bacterial endotoxin test was not interfered. The limitation value of bacterial endotoxin was set as 1. 2 EU/mg. Conclusion It is feasible to detect the bacterial endotoxin of furosemide injection by the bacterial endotoxin test.

  20. Inactivation of Bacterial Spore, Endotoxin, Lipid A, Normal Prion and Abnormal Prion by Exposures to Several Sorts of Gases Plasma.

    Science.gov (United States)

    Shintani, Hideharu

    2016-01-01

    This review discusses the application of several sorts of non-equilibrium gas plasma discharges for sterilization and disinfection treatments against spores or bioburden on/in the healthcare products or biological indicators. The basic properties of electrical discharges are briefly reviewed and thereafter the paper discusses the interactions of gas plasma with several sorts of biological systems such as bacteria, bacterial spores, endotoxins, lipid A and normal and abnormal prion proteins.

  1. 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....

  2. [Effect of bacterial endotoxin on migration of gonadotropin-releasing, hormone producing neurons in rat embryogenesis].

    Science.gov (United States)

    Sharova, V S; Izvol'skaia, M S; Voronova, S N; Zakharova, L A

    2011-01-01

    The effect of bacterial lipopolysaccharide endotoxin (LPS), immune system activator, on differentiation and migration of gonadotropin-releasing, hormone producing neurons in rat embryogenesis has been studied. Intraperitoneal introduction of LPS (18 jg/kg) to pregnant rats on the 12th day of pregnancy led to 50% decrease in total number of GRH-neurons in the forebrain of 17-day-old embryos and 17% decrease in 19-day-old embryos. At the same time, the number of GRH-neurons in the nasal area of the head of 17- and 19-day-old embryos increased by 40 and 50%, respectively, whereas it increased by 20% in olfactory bulbs of 17-day-old embryos and did not changed in olfactory bulbs of 19-day-old embryos. Neither the total number of neurons nor their distribution patterns were affected by the introduction of LPS into pregnant rats on the 15th day of pregnancy. Singular localization of GRH-neurons in embryo forebrain was observed after LPS administration, whereas the neurons were located by groups of 3-4 cells in rostral areas. Therefore, at the early stages of pregnancy, LPS was shown to suppress initial stages of differentiation and migration of GRH producing neurons. The effects observed in our study may be mediated by LPS-induced, proinflammatory cytokines.

  3. 肾康注射液细菌内毒素检查法的建立%Establishment of Bacterial Endotoxin Test of Shenkang Injection

    Institute of Scientific and Technical Information of China (English)

    吴寒寅; 孟德胜

    2011-01-01

    Objective To establish the bacterial endotoxin test of Shenkang injection. Methods Based on Chinese Pharmacopeia (edition 2010, Vol Ⅱ), the interference test and bacterial endotoxin test were conducted on 3 batches of Shenkang injection samples using tachypleus amebocyte lysate from different companies. Results 6- fold dilution of Shenkang Injection had no interference to the bacterial endotoxin test. The limit value of the bacterial endotoxin was L=4 EU/mL. According to the established standard, the results of bacterial endotoxin test in 3 batches of samples were conformed to the required stipulation. Conclusion The established bacterial endotoxin test may be applicable for bacterial endotoxin test of Shenkang injection.%目的 建立肾康注射液的细菌内毒素检查法.方法按2010年版附录Ⅱ细菌内毒素检查法,用不同厂家的鲎试剂对3批肾康注射液分别进行干扰试验和细菌内毒素检查.结果肾康注射液稀释6倍时对细菌内毒素检查无干扰,细菌内毒素限值为4EU/mL.按照拟订标准,3批样品的细菌内毒素检查均符合规定.结论所建立的细菌内毒素检查法可用于肾康注射液的细菌内毒素检查.

  4. 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.

  5. 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.

  6. 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

    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:24D (DD), for 20 days to characterize the rhythms of humoral immunity. Serum alkaline phosphatase (ALP), lysozyme (LYZ), peroxidase (PER) and protease (PRO) exhibited significant rhythmicity under LD but not in DD. No significant rhythms were observed in esterase (ESA) and anti-protease (ANTI) in both photoperiod conditions. Fish reared under LD were subsequently subjected to DD while the group previously under DD was exposed to LD, and this carried on for 3 days before another set of samples was collected. Results revealed that the rhythms of LYZ, PER and PRO but not ALP persisted when photoperiod was changed from LD to DD. Nonetheless, immune parameters remained arrhythmic in the group subjected from DD to LD. Cluster analysis of the humoral immune responses under various light conditions revealed that each photic environment had distinct daily immunological profile. In the second experiment, fish were injected with bacterial endotoxin lipopolysaccharide (LPS) either at ZT3 (day) or at ZT15 (night) to evaluate the temporal sensitivity of humoral immunity to a pathogen-associated molecular pattern. The results demonstrated that responses to LPS were gated by the time of day. LPS significantly modulated serum ALP and ANTI activities but only when the endotoxin was administered at ZT3. Serum LYZ and PER were stimulated at both injection times but with differing response profiles. Modulated LYZ activity was persistent when injected at ZT3 but transient when LPS was applied at ZT15. The magnitude of LPS-induced PER activity was higher when the endotoxin was delivered at ZT3 versus ZT15. It was further shown that plasma cortisol was significantly elevated but only when LPS

  7. 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.

  8. 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.

  9. Determination of endotoxin levels and their impact on interleukin-1 generation in continuous ambulatory peritoneal dialysis and hemodialysis.

    Science.gov (United States)

    Pearson, F C; Dubczak, J; Weary, M; Anderson, J

    1988-01-01

    Endotoxins represent a family of ubiquitous bacterial lipopolysaccharides found in water and raw materials. These substances have the ability to generate interleukin-1 (IL-1) and induce fever, as well as other acute phase phenomena. A study was undertaken to determine levels of background endotoxin in (1) continuous ambulatory peritoneal dialysis solution, (2) spent dialysate subsequent to overnight dwell, (3) hemodialysis solution, and (4) Limulus amebocyte lysate-reactive material (LAL-RM) in hemodialyzers and patient plasma. Levels of endotoxin in all of the above cases were less than thought to be required to induce biological activity, such as pyrogenicity, through IL-1 generation. Although nanogram amounts of LAL-RM are associated with some hollow-fiber membranes as well as the plasma of patients on those membranes, this material per se does not appear to produce IL-1 in vitro.

  10. Establishment of Bacterial Endotoxin Test for Pralidoxime Chloride Injection%氯解磷定注射液细菌内毒素检查法的建立

    Institute of Scientific and Technical Information of China (English)

    王贵英

    2011-01-01

    OBJECTIVE: To establish a method to test bacterial endotoxin in Pralidoxime chloride injection. METHODS: The experiment was carried out in accordance to the bacterial endotoxin test in Chinese Pharmacopoeia (2010 edition). The limit and dilute concentration of bacterial endotoxin were determined. Preliminary interference test, interference test and bacterial endotoxin test of four batches of Pralidoxime chloride injection were performed with tachypleus amebcyte lysate (TAL) manufactured by 2 companies. RESULTS: The limit of bacterial endotoxin was 0.2 EU·mg-1. The bacterial endotoxin was detected with TAL 0.25 EU·mL-1 when the maximum concentration of pralidoxime chloride was 10 mg·rnL-1 without interference. The bacterial endotoxin test of 4 batches of samples was all up to the regulations. CONCLUSION: The results suggest that TAL can be used for the bacterial endotoxin test of Pralidoxime chloride injection.%目的:建立氯解磷定注射液的细菌内毒素检查法.方法:根据《中国药典》2010年版(二部)附录“细菌内毒素检查法”进行.确定样品的细菌内毒素限值和稀释浓度,用2个不同厂家的鲎试剂对4批氯解磷定注射液进行了干扰预试验、干扰试验和细菌内毒素检查.结果:样品细菌内毒素眼值确定为0.2 EU·mg-1,其稀释至10 mg·mL-1及以下浓度时,用灵敏度为0.25 EU·mL-1的鲎试剂检测细菌内毒素,无抑制干扰作用;4批样品细菌内毒素检查均符合规定.结论:氯解磷定注射液可应用鲎试剂进行细菌内毒素检查.

  11. 艾迪注射液细菌内毒素检查方法探讨%Bacterial Endotoxins Test in Aidi Injection

    Institute of Scientific and Technical Information of China (English)

    王丽华

    2011-01-01

    Objective: To establish a method for testing the bacterial endotoxin in Aidi injection by using tachy-pleus amebocyte lysate. Methods: The experiment was implemented according to the loacterial test endotoxins in China Pharmacopoeia (Ⅱ, 2005ed). Results: The results showed that the interference elements could be excluded by diluting glycerol and fructose injection 6 times, and the endotoxin could be tested by amebocyte lysate with accuracy of 0.25 EU/mL. Conclusion: Bacterial endotoxin test is suitable for the detection of bacterial endotoxin in Aidi Injection.%目的:建立艾迪注射液细菌内毒素检查方法.方法:根据《中华人民共和国药典》2005年版Ⅱ部收载的细菌内毒素检查法进行.结果:将艾迪注射液稀释至1/6后可消除干扰因素,用标示灵敏度为0.25 EU/mL的鲎试剂检测细菌内毒素是有效的.结论:细菌内毒素检查法准确、可靠,适用于检测艾迪注射液中的内毒素.

  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. Quantitation of bacterial endotoxin in plasma%血浆中细菌内毒素定量测定方法的研究

    Institute of Scientific and Technical Information of China (English)

    万丽卿; 唐黎明

    2012-01-01

    Objective: To establish a method to quantify the bacterial endotoxin in plasma. Methods: Kinetic -turbidimetric assay was used to determine the concentration and recovery of endotoxin in plasma anticoagulated with three different anticoagulate reagents respectively, and a standard curve of bacterial endotoxin was created by diluting endotoxin with water. In comparison, the other standard curve was created by diluting endotoxin with plasma , amoebocyte lysate was dissolved with antienhancement reagents, and kinetic - turbidimetric assay was used to quantify the endotoxin content and recovery in plasma. Results; Anticoagulate reagents such as heparin lithium and EDTA - K2 interfered with the bacterial endotoxin test, while heparin sodium did not. The standard curve created by diluting endotoxin with diluted and heated human plasma along with amoebocyte lasate dissolved by antienhancement reagents was better at determining bacterial endotoxin in plasma than the standard curve created by diluting endotoxin with water. The standard curve was of good reproducibility, and the endotoxin content in plasma could be quantified accurately. The endotoxin content in colorectal cancer patients was much higher than that in human plasma of healthy people. Conclusion; This kinetic - turbidimetric assay requires the establishment of a standard regression curve by diluting endotoxin with diluted and heated plasma which was anticoagulated by heparin sodium. Together with antienhancement reagents, this assay can quantify the endotoxin concentration in human plasma effectively.%目的:建立定量测定血浆中细菌内毒素的实验方法.方法:以内毒素检查用水制备标准曲线,动态浊度法定量测定分别采用3种抗凝剂的抗凝血中细菌内毒素的含量及回收率;以正常人血浆制备内毒素标准曲线,以抗增液复溶鲎试剂,动态浊度法定量测定人血浆中内毒素的含量及回收率.结果:使用肝素钠抗凝的血浆溶液对内毒

  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. 长春西汀原料药细菌内毒素检查法研究%Study on Bacterial Endotoxin Test of Vinpocetine Raw Material

    Institute of Scientific and Technical Information of China (English)

    周继春

    2015-01-01

    目的:建立长春西汀原料药细菌内毒素检查方法。方法:按2010年版《中国药典》(二部)附录ⅪE细菌内毒素检查法,采用两个厂家的鲎试剂(TAL),对不同批号的样品进行了干扰试验和细菌内毒素检查。结果:高质量浓度长春西汀溶液对TAL与细菌内毒素的凝集反应有干扰作用,经稀释后可排除干扰。结论:本品不干扰质量浓度为0.25 mg/ml,可采用细菌内毒素检查法进行质量控制。%OBJECTIVE:To establish a method for the bacterial endotoxin test of vinpocetine raw material. METHODS:Ac-cording to the bacterial endotoxin test in the Chinese Pharmacopoeia(2010 edition,Ⅱ)Appendix Ⅺ E,the samples with different batches were used for interference test and bacterial endotoxin test by tachypleus amebocyte lysate from 2 manufacturers. RE-SULTS:The vinpocetine solutions with high concentration have interference effect on the agglutination reaction of bacterial endotox-in and the interference can be eliminated by diluting. CONCLUSIONS:The non-interference concentration of vinpocetine solutions is 0.25 mg/ml. Bacterial endotoxin test can be used to control the quality.

  16. Purified monomeric ligand.MD-2 complexes reveal molecular and structural requirements for activation and antagonism of TLR4 by Gram-negative bacterial endotoxins.

    Science.gov (United States)

    Gioannini, Theresa L; Teghanemt, Athmane; Zhang, DeSheng; Esparza, Gregory; Yu, Liping; Weiss, Jerrold

    2014-08-01

    A major focus of work in our laboratory concerns the molecular mechanisms and structural bases of Gram-negative bacterial endotoxin recognition by host (e.g., human) endotoxin-recognition proteins that mediate and/or regulate activation of Toll-like receptor (TLR) 4. Here, we review studies of wild-type and variant monomeric endotoxin.MD-2 complexes first produced and characterized in our laboratories. These purified complexes have provided unique experimental reagents, revealing both quantitative and qualitative determinants of TLR4 activation and antagonism. This review is dedicated to the memory of Dr. Theresa L. Gioannini (1949-2014) who played a central role in many of the studies and discoveries that are reviewed.

  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. 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.

  19. Carnitine deprivation adversely affects cardiovascular response to bacterial endotoxin (LPS) in the anesthetized neonatal pig.

    Science.gov (United States)

    Penn, D; Zhang, L; Bobrowski, P J; Quinn, M; Liu, X; McDonough, K H

    1998-11-01

    Sepsis and endotoxemia are important stressors for the neonate. Newborn infants receiving total parenteral nutrition are routinely deprived of carnitine. To investigate whether carnitine deprivation affects the neonate's ability to respond to endotoxin, 19 newborn piglets received parenteral nutrition for 2-3 weeks that was either carnitine free (CARN-) or supplemented (CARN+) with L-carnitine (400 mg/L). Cardiovascular performance, i.e., heart rate; blood pressure (BP); cardiac output (CO); systemic vascular resistance (SVR), and metabolic response, i.e., plasma glucose; lactate; tumor necrosis factor alpha; tissue nitric oxide; and urinary nitrites, were studied serially in anesthetized piglets for 3 h after endotoxin (lipopolysaccharide (LPS), 250 microg/kg intravenous bolus) or vehicle administration. Plasma and tissue carnitine values were lower in CARN- than in CARN+ piglets. Prior to LPS, no differences were found for most parameters (excepting lower diastolic BP and SVR in CARN- animals). Systolic, diastolic, and mean BP fell after LPS but recovered by the end of the experiment. Nadirs were lower in CARN- than in CARN+ piglets. CO tended to be higher in CARN- than in CARN+ animals and fell after LPS. SVR fell after LPS and was lower in CARN- than in CARN+ piglets. LPS-treated animals transiently increased urinary flow. By all measures (plasma tumor necrosis factor alpha, glucose and lactate, tissue nitric oxide, and urinary nitrite excretion), LPS provocation was similar for both groups. Chronologically, BP changes were more closely related to SVR than to CO. Our findings suggest that carnitine deprivation diminishes tissue carnitine concentrations and adversely affects cardiovascular response to LPS, in part mediated by the peripheral vasculature.

  20. Short-chain inulin-like fructans reduce endotoxin and bacterial translocations and attenuate development of TNBS-induced colitis in rats.

    Science.gov (United States)

    Ito, Hiroyuki; Tanabe, Hiroki; Kawagishi, Hirokazu; Tadashi, Wada; Yasuhiko, Tomono; Sugiyama, Kimio; Kiriyama, Shuhachi; Morita, Tatsuya

    2009-10-01

    Anti-inflammatory effects of short-chain inulin-like fructans (SCF) on trinitrobenzene sulfonic acid (TNBS)-induced colitis were investigated in rats, focusing specifically on endotoxin and bacterial translocations. SCF with degrees of polymerization (DP) of 4 and 8 were used. Rats were fed either control diet or diets including 60 g DP4 or DP8 per kilogram for 7 days, and then received intracolonic TNBS and were fed the respective diets for a further 10 days. DP4 and DP8 significantly reduced colonic injuries as assessed by damage score, but the reduction of colonic myeloperoxidase activity was manifest solely with DP8. At 3 days after colitis induction, bacterial translocation to the mesenteric lymph node was significantly lower in the DP4 and DP8 groups, but significant reduction in the portal endotoxin concentration was achieved solely in the DP8 group. Immediately prior to colitis induction, cecal immunoglobulin A and mucin concentrations were higher in the DP4 and DP8 groups, but these changes were abolished at 10 days post colitis induction. The data suggest that SCF exert prophylactic effects against TNBS colitis, presumably as a result of inhibitory effects on endotoxin and bacterial translocations.

  1. Study on Bacterial Endotoxin Test of Cimetidine Raw Material%西咪替丁原料药细菌内毒素检查方法的探讨

    Institute of Scientific and Technical Information of China (English)

    金艳; 王益民

    2012-01-01

    Objective To establish a bacterial endotoxin test for cimetidine raw material. Methods According to the method of the bacteria endotoxin test in the Chinese Pharmacopoeia (edition 2010, Vol. II). Results When the mass concentration of sample was diluted to 4.0 mg/mL,no interference effect on the reaction of bacterial endotoxin and TAL was found. The limit value of the bacterial endotoxin was 0. 5 EU/mg. Conclusion It is feasible to detect the bacterial endotoxin of cimetidine raw material by the method of bacterial endotoxin test.%目的 建立西咪替丁原料药的细菌内毒素检查方法.方法 参照2010年版细菌内毒素检查法进行试验.结果 样品质量浓度稀释至4.0 g/L时,对鲎试剂与细菌内毒素的反应无干扰作用.其细菌内毒素限值定为0.5 EU/mg.结论 西咪替丁原料药采用内毒素检查法(鲎试剂法)是可行的.

  2. Current trends in endotoxin detection and analysis of endotoxin-protein interactions.

    Science.gov (United States)

    Dullah, Elvina Clarie; Ongkudon, Clarence M

    2017-03-01

    Endotoxin is a type of pyrogen that can be found in Gram-negative bacteria. Endotoxin can form a stable interaction with other biomolecules thus making its removal difficult especially during the production of biopharmaceutical drugs. The prevention of endotoxins from contaminating biopharmaceutical products is paramount as endotoxin contamination, even in small quantities, can result in fever, inflammation, sepsis, tissue damage and even lead to death. Highly sensitive and accurate detection of endotoxins are keys in the development of biopharmaceutical products derived from Gram-negative bacteria. It will facilitate the study of the intermolecular interaction of an endotoxin with other biomolecules, hence the selection of appropriate endotoxin removal strategies. Currently, most researchers rely on the conventional LAL-based endotoxin detection method. However, new methods have been and are being developed to overcome the problems associated with the LAL-based method. This review paper highlights the current research trends in endotoxin detection from conventional methods to newly developed biosensors. Additionally, it also provides an overview of the use of electron microscopy, dynamic light scattering (DLS), fluorescence resonance energy transfer (FRET) and docking programs in the endotoxin-protein analysis.

  3. Dynamic Monitoring on the Bacterial Endotoxin Concentration in Whole Blood and Blood Component%全血及成分血内毒素动态监测

    Institute of Scientific and Technical Information of China (English)

    鲍自谦; 刘怡; 王飞; 陈云龙

    2012-01-01

    目的 通过监测全血及成分血的内毒素含量动态变化,探讨内毒素监控的意义,为评估血液安全提供依据,保证安全输血.方法 选择2009年3月至4月本血液中心献血者中,符合国家《献血者健康检查要求》的自愿献血者血液样品300份为研究对象.按血液成分分为3组:全血组(n=50)、悬浮RBC成分血组(n=150)及PLT成分血组(n=100).采用动态浊度法内毒素定量检测技术分别于各组血液保存前期、中期及后期检测各个血液样品内毒素含量,并采用方差分析比较各个检测时间点血样内毒素变化情况.结果 全血保存期前中后期中,内毒素含量在部分时段差异有统计学意义(W=0.860,P<0.05);而PLT悬液、悬浮RBC在3个时间段的内毒素含量差异无统计学意义(P>0.05).结论 对全血及成分血的内毒素含量监测,结果显示血液保存期的内毒素含量稳定与其细菌培养阴性相吻合,表明现行的采供血流程的质量控制是有效的,可有效保障血液的安全.%Objective To explore the importance of dynamic monitoring on the bacterial endotoxin and assess the blood safety to guarantee the safety of blood transfusion,through dynamically monitoring the bacterial endotoxin concentration in whole blood and blood component. Methods From March to April 2009,300 donors' samples,which had met the requirements of donors' health check,were included in this study.According to the blood component,all these samples were performed into three groups:whole blood group (n=50),red blood cells (RBC) suspension group (n=150),platelet (PLT) components group (n=100).Dynamic turbidity in blood which were respectively stored at early stage,metaphase and later stage,were used as the detection quantitative method of bacterial endotoxin.And the results were further analyzed by variance of repeated measurement data. Results There was no difference of the bacterial endotoxin concentration during the preservation in

  4. 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.

  5. Endotoxin contamination delays the foreign body reaction

    NARCIS (Netherlands)

    van Putten, Sander M.; Wubben, Maike; Plantinga, Josee A.; Hennink, Wim E.; van Luyn, Marja J. A.; Harmsen, Martin C.

    2011-01-01

    Biomaterials are at continuous risk of bacterial contamination during production and application. In vivo, bacterial contamination of biomaterials delays the foreign body reaction (FBR). Endotoxins such as lipopolysaccharides (LPS), major constituents of the bacterial cell wall, are potent stimulato

  6. 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.

  7. 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.

  8. Content Determination of Bacterial Endotoxin in Docetaxel Injection by Kinetic Turbidimetric Method%动态浊度法测定多西他赛注射液中细菌内毒素的含量

    Institute of Scientific and Technical Information of China (English)

    祝清芬; 魏霞

    2011-01-01

    OBJECTIVE: To establishment a method for determine the concentration of bacterial endotoxin in Docetaxel injection. METHODS: Kinetic turbidimetric method was adopted to generate the standard curve and dilution rate of test sample. Bacterial endotoxin was added into test sample for interference test. Linear range and bacterial endotoxin in samples were determined. RESULTS: The linear range of bacterial endotoxin was 0.031 25~2.0 EU·mL-1 (r=-0.999 9). The endotoxin test wasn't interfered by 12~96 times diluted solution. The recovery rate of bacterial endotoxin was 50% ~200%. The concentration of endotoxin in the 3 batches of samples met the requirements for bacterial endotoxins. CONCLUSION: The kinetic turbidimetric method is suitable for bacterial endotoxin test of Docetaxei injection.%目的:建立测定多西他赛注射液中细菌内毒素含量的方法.方法:采用动态浊度法.确立细茵内毒素检查的标准曲线及稀释倍数,通过测定供试液中外加内毒素的回收率进行干扰试验,确定样品检测浓度线性范围,并定量测定样品中的细茵内毒素.结果:内毒素检测浓度线性范围为0.031 25~2.0 EU·mL-1(r=-0.999 9);样品在稀释12~96倍时对试验无干扰作用;细菌内毒素回收率为50%~200%;3批样品的内毒素定量测定结果符合规定.结论:动态浊度法可用于多西他赛注射液中细茵内毒素的定量检测.

  9. Content Determination of Bacterial Endotoxin in Gatifloxacin by Kinetic Chromogenic Analysis%动态显色法定量测定加替沙星原料药中细菌内毒素含量

    Institute of Scientific and Technical Information of China (English)

    夏博; 郭锋

    2015-01-01

    Objective:To establish a method for the detection of bacterial endotoxins in gatifloxacin.Methods:Kinetic Chromogenic analysis was adopted to generate the standard curve and dilution rate of test sample.Bacterial endotoxin was added into test sample for in-terference test, and bacterial endotoxins in samples were determined.Results:The endotoxin test wasn′t interfered at concentrations be-low 0.312 mg・mL-1 .The recovery rate of bacterial endotoxin was 50%~200%.The concentration of endotoxin in the 3 batches of sam-ples met the requirements for bacterial endotoxins.Conlusion:The kinetic chromogenic analysis is suitable for bacterial endotoxin test of iodixanol injection.%目的:应用动态显色法定量测定加替沙星中的细菌内毒素含量。方法:建立动态显色法测定细菌内毒素标准曲线,通过外加内毒素测回收试验,对样品预处理方法进行了验证,证实供试液制备过程不会破坏样品本底的细菌内毒素;通过干扰试验确定样品浓度检测范围,并对样品的细菌内毒素进行定量测定。结果:样品在0.312 mg・mL-1以下浓度时,对试验无干扰作用,细菌内毒素回收率在50%~200%内,样品中的内毒素含量可定量测定。结论:动态显色法可用于定量检测加替沙星中的细菌内毒素。

  10. 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 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 were

  11. 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

  12. 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.

  13. Alfalfa pellet-induced subacute ruminal acidosis in dairy cows increases bacterial endotoxin in the rumen without causing inflammation.

    Science.gov (United States)

    Khafipour, E; Krause, D O; Plaizier, J C

    2009-04-01

    A study was conducted to determine if subacute ruminal acidosis (SARA) induced by feeding alfalfa pellets results in increases in free bacterial lipopolysaccharide (LPS) in rumen fluid and peripheral blood, and acute phase proteins in plasma, and to determine the effect of alfalfa pellet-induced SARA on feed intake, rumen fermentation characteristics, milk production and composition, and blood metabolites. Eight lactating Holstein cows, 4 of which were ruminally cannulated, were used in a 6-wk experiment and were fed once daily at 0900 h. During wk 1, cows received a diet containing 50% of DM as concentrate and 50% of DM chopped alfalfa hay. Between wk 2 and wk 6, alfalfa hay was gradually replaced with alfalfa pellets at the rate of 8% per week to reduce rumen pH. Rumen pH was monitored continuously in the ruminally cannulated cows using indwelling pH probes. Rumen fluid and peripheral blood were sampled 15 min before feed delivery and at 6 h after feed delivery. Based on adopted threshold of SARA of at least 180 min/d below pH 5.6, SARA was induced from wk 3 onwards. Replacing 40% of alfalfa hay with alfalfa pellets quadratically increased the DMI from 18.1 kg/d in wk 1 to 23.4 kg/d in wk 6. This replacement linearly decreased milk yield (32.7 vs. 35.9 kg/d) and milk fat percentage and yield (2.32 vs. 3.22%, and 0.77 vs. 1.14 kg/d, respectively), but increased milk protein percentage and yield (3.80 vs. 3.04%, and 1.23 vs. 1.07 kg/d, respectively). This gradual replacement also linearly increased the daily averages of total volatile fatty acids (90 to 121.9 mM), acetate (53.9 to 66.8 mM), propionate (21.5 to 39.6 mM), and osmolality (277.7 to 293.8 mmol/kg) in the rumen and decreased the acetate to propionate ratio from 2.62 to 1.73. Replacing alfalfa hay with alfalfa pellets linearly increased blood lactate from 1.00 mM in wk 1 to a peak of 3.46 mM in wk 5. Induction of SARA in this study increased free rumen LPS concentration from 42,122 endotoxin unit (EU

  14. Detection of bacterial endotoxin in milrinone injection by the method of kinetic turbidimetric limulus test%米力农注射液动态浊度法细菌内毒素检查方法研究

    Institute of Scientific and Technical Information of China (English)

    祝清芬; 国明; 魏霞; 李涛

    2013-01-01

    Objective: To revise the criterion for bacterial endotoxins test of milrinone injection, and to measure the concentration of endotoxin in milrinone injection by kinetic turbidimetric technique. Methods:The limit for bacterial endotoxin in this product was set according to ChP 2010. Endotoxin solutions of four concentrations were prepared to generate the standard curve. The interfering factors test was done by measuring the concentration of the endotoxin added to the sample solution, i. e. , the recovery of added endotoxin. The concentration of endotoxin in the sample solution was measured. Results:The correlation coefficient was 0.996 2, which indicated that the standard curve was valid because the correlation coefficient must be greater than or equal to 0. 980 according to ChP 2010. The recovery of the added endotoxin in the sample solution was within 50% to 200% when the sample was diluted to 8 times or more of its original volume. The measured concentration of endotoxin in the sample met the requirement for bacterial endotoxin. Conclusion: The bacterial endotoxin test method for milrinone injection has been revised. The kinetic turbidimetric technique is suitable for bacterial endotoxin test of this product.%目的:修订米力农注射液细菌内毒素检查法质量标准,并采用动态浊度法对其细菌内毒素检查进行方法学研究.方法:按《中华人民共和国药典》2010年版二部,修改本品细菌内毒素限值.使用动态浊度法,研究米力农注射液对细菌内毒素检查试验的干扰情况,并定量测定样品中细菌内毒素含量.结果:标准曲线回归方程相关系数绝对值为0.996 2(应≥0.980),标准曲线成立.本品在8倍稀释时细菌内毒素回收率在50%~200%范围内,表明此稀释倍数对细菌内毒素检查法无干扰作用,1 1批样品中的内毒素定量测定结果符合修订后的标准.结论:修订了本品细菌内毒素检查法质量标准,本品可采用动态浊度法进行细菌内毒素检查.

  15. 醒脑静注射液细菌内毒素检查法研究%The study for the bacterial endotoxins determination method of the Xingnaojing injection

    Institute of Scientific and Technical Information of China (English)

    匡翠莲; 王璐; 张红宇

    2012-01-01

      目的:建立醒脑静注射液细菌内毒素检查方法.方法:按中国药典2005年版二部附录ⅪE收载的细菌内毒素检查方法进行试验和结果判断,计算醒脑静注射液的细菌内毒素限值.结果:醒脑静注射液的细菌内毒素限值为2.5EU•mg.结论:可建立醒脑静注射液的细菌内毒素检查方法.%  Objective: To set up the bacterial endotoxins determination method for the Xingnaojing injection. Methods: The experiment was carried on and the result was judged according to the appendix ⅪE of Chinese Pharmacopoeia 2005 Vol. Ⅱ. To calculate bacterial endotoxin limit of the Xingnaojing injection. Results: Bacterial endotoxins limit of XingNaojing injection was 2.5EU•mg. Conclusions:The established method of endotoxins test was feasible for XingNaojing injection.

  16. Study on Dynamic Turbidimetry of Bacterial Endotoxin Test for Fasudil Hydrochloride%动态比浊法检测盐酸法舒地尔注射剂细菌内毒素的含量

    Institute of Scientific and Technical Information of China (English)

    李展; 张晗; 周继春; 杨海燕

    2015-01-01

    目的:建立盐酸法舒地尔注射剂细菌内毒素动态浊度法检测方法。方法:采用动态浊度法进行干扰试验。结果:3.75 mg·mL-1盐酸法舒地尔注射剂无干扰作用。结论:本品可用动态浊度法进行细菌内毒素定量检测,其内毒素限值可定为每1 mg盐酸法舒地尔注射剂中含内毒素的量应小于5.0 EU。%Objective: To establish dynamic turbidimetry of bacterial endotoxin test for fasudil hydrochloride injection.Methods:Dynamic turbidimetry was used for the interference test for fasudil hydrochloride injection. Results:Fasudil hydrochloride injection had no interference effects on the reaction of tachypleus amebocyte lysate (TAL) at the concentration of 3.75 mg·mL-1.Conclusion:Dynamic turbidimetry of bacterial endotoxin test can be adopted for a quantitative test of endotoxin. The limit of bacterial endotoxin in fasudil hydrochloride injection should be set less than 5.0 EU·mg-1.

  17. Short communication: Ovine leukocyte telomere length is associated with variation in the cortisol response to systemic bacterial endotoxin challenge.

    Science.gov (United States)

    Yip, L; Oh, S Y; Li, Z; You, Q; Quinton, V M; Gilchrist, G C; Karrow, N A

    2016-04-01

    Stress has been associated with biological aging and numerous age-related diseases. This may be due, in part, to accelerated shortening of telomeres, which are critical genomic structures that cap and protect chromosomal ends. Dysfunction of the hypothalamic-pituitary-adrenal axis may indirectly contribute to telomere shortening if an animal reacts too strongly or weakly to a stressor, leading to accelerated biological aging. In this study, outbred Rideau-Arcott sheep were stress challenged with Escherichia coli endotoxin and classified as high, middle, or low cortisol responders to investigate a potential relationship between cortisol response and age, and telomere length. In the present study, no association was found between age and telomere length. The study, however, revealed shorter telomeres in high and low cortisol responders compared with the middle cortisol responders, which suggests that health and longevity may be compromised in extreme high- and low-stress-responding sheep.

  18. Determination of Interference During In Vitro Pyrogen Detection: Development and Characterization of a Cell-Based Assay.

    Science.gov (United States)

    Palma, Linda; Rossetti, Francesca; Dominici, Sabrina; Buondelmonte, Costantina; Rocchi, Marco B L; Rizzardi, Gian P; Vallanti, Giuliana; Magnani, Mauro

    2016-12-20

    Contamination of pharmaceutical products and medical devices with pyrogens such as endotoxins is the most common cause of systemic inflammation and, in worst cases, of septic shock. Thus, quantification of pyrogens is crucial. The limulus amebocyte lysate (LAL)-based assays are the reference tests for in vitro endotoxin detection, in association with the in vivo rabbit pyrogen test (RPT), according to European Pharmacopoeia (EP 2.6.14), and U.S. Pharmacopoeia (USP ). However, several substances interfere with LAL assay, while RPT is not accurate, not quantitative, and raises ethical limits. Biological assays, as monocyte activation tests, have been developed and included in European Pharmacopoeia (EP 7.0; 04/2010:20630) guidelines as an alternative to RPT and proved relevant to the febrile reaction in vivo. Because this reaction is carried out by endogenous mediators under the transcriptional control of nuclear factor-kappaB (NF-kappaB), we sought to determine whether a NF-kappaB reporter-gene assay, based on MonoMac-6 (MM6) cells, could reconcile the basic mechanism of innate immune response with the relevance of monocytoid cell lines to the organism reaction to endotoxins. This article describes both optimization and characterization of the reporter cells-based assay, which overall proved the linearity, accuracy, and precision of the test, and demonstrated the sensitivity of the assay to 0.24 EU/mL endotoxin, close to the pyrogenic threshold in humans. Moreover, the assay was experimentally compared to the LAL test in the evaluation of selected interfering samples. The good performance of the MM6 reporter test demonstrates the suitability of this assay to evaluate interfering or false-positive samples.

  19. Verification of the bacterial endotoxin inspection method of vinorelbine tartrate%酒石酸长春瑞滨的细菌内毒素检查方法验证

    Institute of Scientific and Technical Information of China (English)

    嵇海澄; 王倜; 王春玲; 李孝壁

    2014-01-01

    目的:参考 USP、EP,探讨并建立酒石酸长春瑞滨的细菌内毒素标准与检查方法(凝胶法)。方法按《中国药典》(2010年版)二部附录Ⅺ E 细菌内毒素检查方法进行。结果经验证,酒石酸长春瑞滨的细菌内毒素限值确定为 L <2.0IU/mg(以长春瑞滨计),通过稀释法可消除干扰,无干扰浓度为0.035mg/mL(以长春瑞滨计)。结论可按此标准采用本法对本品进行细菌内毒素检查。%Objective To explore and establish the standard and inspection method (gel) of vinorelbine tartrate’ bacterial endotoxin refer to USP, EP. Methods According to the“Chinese Pharmacopoeia”(2010 Edition) two appendix XIE’bacterial endotoxin inspection method. Results Through verification, the vinorelbine tartrate’ bacterial endotoxin limit values determined L < 2.0IU/mg (taken Changchun vinorelbine meter), and eliminated interference by dilution method, no interference concentration was 0.035mg/mL(taken Changchun vinorelbine meter). Conclusion We can make the bacterial endotoxin test according to this standard by adopting the method for this product.

  20. 多烯磷脂酰胆碱注射液细菌内毒素检查法的建立%Detection of Bacterial Endotoxins in Polyene Phosphatidylcholine Injection

    Institute of Scientific and Technical Information of China (English)

    王宗春; 郭咸希; 张洪

    2011-01-01

    目的:建立多烯磷脂酰胆碱注射液的细菌内毒素检查法.方法:按照<中国药典)2010年版(二部)附录ⅪE细菌内毒素检查法进行干扰试验.结果:多烯磷脂酰胆碱注射液经40倍稀释后,用灵敏度λ为0.25 EU·ml-1的鲎试剂进行试验,供试品不干扰细菌内毒素检查.结论:鲎试剂方法用于多烯磷脂酰胆碱注射液细菌内毒素检查可行.%Objective: To establish a method for the detection of bacterial endotoxin in polyene phosphatidylcholine injection.Method: The interference test was carried out according to the guideline and procedure specified in appendix Ⅺ E, volume two, Chinese Pharmacopoeia 2010 edition. Result: By using limulus agent with sensitivity of 0. 25 EU· ml- 1, the sample did not interfere with the detection of bacterial endotoxin when it was diluted 40 times. Conclusion: The established method of bacterial endotoxins test for polyene phosphatidylcholine injection is feasible.

  1. 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.

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

    Science.gov (United States)

    Haskå, G; Nystrand, R

    1979-12-01

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

  3. Aplicabilidade do Teste de Ativação de Monócitos (MAT no Brasil: importância da sua utilização como teste para detecção de pirogênios no controle da qualidade de produtos injetáveis | Applicability of the Monocyte Activation Test (MAT in Brazil: the importance of its use as a test for the detection of pyrogens in the quality control of injectable products

    Directory of Open Access Journals (Sweden)

    Cristiane Caldeira da Silva

    2015-08-01

    kick-off study for future studies. Products’ monographs in pharmacopeias in the United States, Europe, and Brazil that are recommended for RPT or Bacterial Endotoxins Test (BET are as follows. RPT: 20 in American, 37 in European, and 28 in Brazilian pharmacopoeias. BET: 619 in American, 157 in European, and 41 in Brazilian pharmacopoeias. Four products require pyrogenicity testing in three pharmacopoeias. Both RPT and BET are recommended by six monographs in Brazilian pharmacopeias and 15 in European pharmacopoeias. In Brazilian pharmacopoeias most are biological. These products should be the first to be tested for their applicability to MAT because they are currently tested mainly by RPT.

  4. 鲎试剂检测复方柳安咖注射液中细菌内毒素的可行性研究%Feasibility study on bacterial endotoxin of compound sodium salicylate antipyrine and caffeine injection with the TAL

    Institute of Scientific and Technical Information of China (English)

    赵祎; 张红宇; 王莉; 李海山

    2015-01-01

    Objective To establish a method for the bacterial endotoxin test of compound sodium salicylate antipyrine and caffeine injection.Method The bacterial endotoxin test was carried out according to the appendix XI E of Chinese Pharmacopoeia (Vol.Ⅱ, edition 2010).Result The compound sodium salicylate antipyrine and caffeine injection by diluting to a concentration of more than eighty times was noninterference for bacterial endotoxin test of gel -clot.Conclusion The bacterial endotoxins test is suitable for the detection of endotoxin in the compound sodium salicylate antipyrine and caffeine injection.%目的:通过对复方柳安咖注射液进行细菌内毒素检查法研究,建立复方柳安咖注射液细菌内毒素检查方法。方法:采用2010年版《中国药典》(二部)附录 XI E 细菌内毒素检查法进行。结果:复方柳安咖注射液经80倍及以上稀释,对细菌内毒素检查法(凝胶法)无干扰作用。结论:细菌内毒素检查法适用于复方柳安咖注射液中内毒素的检测。

  5. International Conference on Harmonisation; Guidance on Q4B Evaluation and Recommendation of Pharmacopoeial Texts for Use in the International Conference on Harmonisation Regions; Annex 14 on Bacterial Endotoxins Test General Chapter; availability. Notice.

    Science.gov (United States)

    2013-10-23

    The Food and Drug Administration (FDA) is announcing the availability of a guidance entitled "Q4B Evaluation and Recommendation of Pharmacopoeial Texts for Use in the International Conference on Harmonisation Regions; Annex 14: Bacterial Endotoxins Test General Chapter.'' The guidance was prepared under the auspices of the International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH). The guidance provides the results of the ICH Q4B evaluation of the Bacterial Endotoxins Test General Chapter harmonized text from each of the three pharmacopoeias (United States, European, and Japanese) represented by the Pharmacopoeial Discussion Group (PDG). The guidance conveys recognition of the three pharmacopoeial methods by the three ICH regulatory regions and provides specific information regarding the recognition. The guidance is intended to recognize the interchangeability between the local regional pharmacopoeias, thus avoiding redundant testing in favor of a common testing strategy in each regulatory region. The guidance is in the form of an annex to the core guidance on the Q4B process entitled "Q4B Evaluation and Recommendation of Pharmacopoeial Texts for Use in the ICH Regions (core ICH Q4B guidance).

  6. 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.

  7. 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

  8. 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 endotoxin contamination and control in treated water following each unit of the WTP processes and its SW from reservoirs are discussed and compared with regard to bacterial cell counts and particle characteristics, which were dependent, to a certain extent, on different flow rates and turbulence of the water environments.

  9. Endotoxin-like activity associated with Lyme disease Borrelia.

    Science.gov (United States)

    Fumarola, D; Munno, I; Marcuccio, C; Miragliotta, G

    1986-12-01

    The newly recognized spirochete, Borrelia burgdorferi, the causative agent of Lyme Disease, has been examined for endotoxin-like activities as measured by the standard Farmacopea Ufficiale della Republica Italiana rabbit fever test and the Limulus amoebocyte lysate assay. The suspension of heat-killed microorganism caused a febrile response at a dose of 1 X 10(8) bacteria pro kilo. Similar results were obtained in the Limulus assay where the heat-killed spirochetes stimulated formation of solid clot until the concentration of 1 X 10(5) per ml. Both in pyrogen test and in Limulus assay heat-killed Escherichia coli exhibited a higher degree of potency. These results show that LD-Borrelia possess endotoxin-like activities which could help in understanding the pathogenesis of the clinical symptomatology of the disease.

  10. 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.

  11. 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)

  12. 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.

  13. The natural immunity to evolutionary atavistic endotoxin for human cancer.

    Science.gov (United States)

    Moncevičiūtė-Eringienė, Elena; Rotkevič, Kristina; Grikienis, Ruta Grikienyte

    2015-11-01

    We propose a new theory of the immunological control of cancer corresponding to the hypothesis that the specific natural immunity to evolutionary atavistic endotoxin has a potential role in human cancer prevention. The results of our studies have shown that IgMNAE, i.e. endogenous or spontaneous IgM class antibodies to enterobacterial lipopolysaccharide molecules (lipid A), control the immune mechanisms responsible for the internal medium stability not only against the damaging impact of the carcinogenic factors, but also against the malignant transformation of its own degenerated cells. Among people who in 1979 and 1982 had IgMNAE in their blood serum, after 15-30years fell ill with cancer 10%, versus 15% among people who had no IgMNAE (pimmunity to endotoxin it is possible to see the formation of the respective evolutionary protective reactions which protect the damaged cells from acquiring resistance to damaging factors and thus from becoming an independent new parasitic population. Thereby the presented theory of the immunological control of cancer has a causal connection with our evolutionary resistance theory of the origin of cancer. Collectively, these data suggest that activation of natural immunity to endotoxin and production of vaccines against evolutionary atavistic endotoxin or gram-negative bacterial endotoxin can be helpful when applied in cancer prophylaxis for persons with a low level of natural immunity to endotoxin and perhaps in creating immunotherapeutic methods for stopping the endogenous parasitism of tumour cells by binding IgMNAE to atavistic endotoxin in cancer patients.

  14. Comparison of the effects of ketoprofen and flunixin meglumine on the in vitro response of equine peripheral blood monocytes to bacterial endotoxin.

    Science.gov (United States)

    Jackman, B R; Moore, J N; Barton, M H; Morris, D D

    1994-04-01

    The purpose of this study was to investigate the in vitro effects of flunixin meglumine, a cyclo-oxygenase inhibitor, and ketoprofen, a reported cyclo-oxygenase and lipoxygenase inhibitor, on the synthesis of cyclo-oxygenase end-products thromboxane B2 and prostaglandin E2, lipoxygenase derived 12-hydroxyeicosatetraenoic acid, tumor necrosis factor and tissue factor. Six adult horses were each randomly administered flunixin meglumine (1.1 mg/kg) or ketoprofen (2.2 mg/kg) intravenously every 12 hours with the drug treatments separated by two weeks. Blood samples were obtained prior to initiating treatment, the last day of treatment and for two consecutive days after the termination of treatment for measurement of serum concentrations of thromboxane B2 as well as isolation of peripheral blood monocytes. Quantitation of unstimulated, endotoxin- and calcium ionophore-induced synthesis of thromboxane B2, prostaglandin E2, 12-hydroxyeicosatetraenoic acid, tumor necrosis factor and tissue factor by peripheral blood monocytes was performed in vitro. Both flunixin meglumine and ketoprofen significantly decreased serum concentrations of thromboxane B2 demonstrating in vivo cyclo-oxygenase inhibition. There were no significant differences between drug treatment groups in the in vitro production of thromboxane B2, prostaglandin E2, 12-hydroxy-eicosatetraenoic acid, tumor necrosis factor or tissue factor. This study does not identify significant differences between the effects of flunixin meglumine and ketoprofen.

  15. Endotoxin and tumor necrosis factor-receptor levels in portal and hepatic vein of patients with alcoholic liver cirrhosis receiving elective transjugular intrahepatic portosystemic shunt

    DEFF Research Database (Denmark)

    Trebicka, Jonel; Krag, Aleksander; Gansweid, Stefan

    2011-01-01

    In cirrhosis portal hypertension can promote bacterial translocation and increase serum endotoxin levels. Vice versa, endotoxin aggravates portal hypertension by induction of systemic and splanchnic vasodilation, and by triggering hepatic inflammatory response via tumor necrosis factor α (TNFα......). However, the hepatic elimination of endotoxin in cirrhotic patients with severe portal hypertension, in the absence of acute complications, has not been investigated so far....

  16. Standard Molded Composite Rocket Pyrogen Igniter - A progress report

    Science.gov (United States)

    Lucy, M. H.

    1978-01-01

    The pyrogen igniter has the function to furnish a controlled, high temperature, high pressure gas to ignite solid propellant surfaces in a rocket motor. Present pyrogens consist of numerous inert components. The Standard Molded Pyrogen Igniter (SMPI) consists of three basic parts, a cap with several integrally molded features, an ignition pellet retainer plate, and a tube with additional integrally molded features. A description is presented of an investigation which indicates that the SMPI concept is a viable approach to the design and manufacture of pyrogen igniters for solid propellant rocket motors. For some applications, combining the structural and thermal properties of molded composites can result in the manufacture of lighter assemblies at considerable cost reduction. It is demonstrated that high strength, thin walled tubes with high length to diameter ratios can be fabricated from reinforced plastic molding compound using the displacement compression process.

  17. Adherent endotoxin on dental implant surfaces: a reappraisal.

    Science.gov (United States)

    Morra, Marco; Cassinelli, Clara; Bollati, Daniele; Cascardo, Giovanna; Bellanda, Marco

    2015-02-01

    Osteoimmunology is the crosstalk between cells from the immune and skeletal systems, suggesting a role of pro-inflammatory cytokines in the stimulation of osteoclast activity. Endotoxin or bacterial challenges to inflammatory cells are directly relevant to dental implant pathologies involving bone resorption, such as osseointegration failure and peri-implantitis. While the endotoxin amount on implant devices is regulated by standards, it is unknown whether commercially available dental implants elicit different levels of adherent-endotoxin stimulated cytokines. The objective of this work is to develop a model system and evaluate endotoxin-induced expression of pro-inflammatory cytokine genes relevant to osteoclast activation on commercially available dental implants. Murine J774-A1 macrophages were cultured on Ti disks with different level of lipopolysaccharide (LPS) contamination to define the time-course of the inflammatory response to endotoxin, as evaluated by reverse transcription polymerase chain reaction analysis. The developed protocol was then used to measure adherent endotoxin on commercially available packaged and sterile dental implants in the "as-implanted" condition. Results show that tested dental implants induce variable expression of endotoxin-stimulated genes, sometimes above the level expected to promote bone resorption in vivo. Results are unaffected by the specific surface treatment; rather, they likely reflect care in cleaning and packaging protocols. In conclusion, expression of genes that enhance osteoclast activity through endotoxin stimulation of inflammatory cells is widely different on commercially available dental implants. A reappraisal of the clinical impact of adherent endotoxins on dental (and bone) implant devices is required in light of increasing knowledge on crosstalk between cells from the immune and skeletal systems.

  18. Endotoxin release from biologically active bench-scale drinking water anthracite/sand filters

    Energy Technology Data Exchange (ETDEWEB)

    Anderson, W.B.; Mayfield, C.I.; Huck, P.M. [University of Waterloo, Waterloo, ON (Canada). Dept. of Civil and Environmental Engineering

    2008-12-15

    Endotoxin release from biologically active dual-media (anthracite/sand) bench-scale drinking water filters was investigated. The biological filters were typically net producers of endotoxin during normal operation with mean concentrations increasing from 6 endotoxin units (EU)/mL to 16 EU/mL in filter influent and effluent, respectively. The filter operating condition resulting in the greatest release of endotoxin occurred upon shutdown and subsequent restart, with the highest observed filter effluent endotoxin concentration being 745 EU/mL in the first pore volume following the return of flow through the filter. Effluent samples collected following filter shutdowns were chlorinated to determine the effect of bacterial cell death on endotoxin release. Chlorination did not induce immediate 'dumping' of endotoxin, nor did holding the chlorinated samples for 5 d result in an increase in aqueous endotoxin concentration. In addition to endotoxin, measurable quantities of potassium were detected in the interstitial water in the biofilter during shutdown periods. In order to reduce potential risk to dialysis patients and humidifier users, it is recommended that, following even short biofilter shutdowns, filters should be immediately backwashed or, failing this, the first five pore volumes of effluent water be discharged to waste to allow endotoxin levels to return to pre-shutdown values.

  19. Endotoxin contamination: a key element in the interpretation of nanosafety studies.

    Science.gov (United States)

    Li, Yang; Boraschi, Diana

    2016-02-01

    The study of toxicity and potential risks of engineered nanoparticles is of particular importance in nanomedicine. Endotoxin, a common contaminant of bacterial origin, has biological effects that can mask the true biological effects of nanoparticles, if its presence is overlooked. In this review, we report the features of nanoparticle contamination by endotoxin, and the different biological effects of endotoxin-contaminated nanoparticles. We will describe different methods for endotoxin detection applied to nanoparticles, and discuss their pros and cons. Eventually, we describe various methods for eliminating endotoxin contamination in nanoparticles. Although there is no universal technique for efficiently removing endotoxin from nanoparticles, specific solutions can be found case by case, which can allow us to perform nanosafety studies in biologically relevant conditions.

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

    Science.gov (United States)

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

  2. [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.

  3. Membrane adsorber for endotoxin removal

    Directory of Open Access Journals (Sweden)

    Karina Moita de Almeida

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

  4. Study on Dynamic Turbidimetry of Bacterial Endotoxin Test for Dexrabeprazole Sodium for Injection%动态比浊法检测注射用右旋雷贝拉唑钠细菌内毒素方法的建立

    Institute of Scientific and Technical Information of China (English)

    李展; 周继春

    2015-01-01

    Objective:To establish a method of dynamic turbidimetry of bacterial endotoxin test for Dexrabeprazole Sodium for Injec-tion.Methods:According to the relevant appendix of Chinese Pharmacopoeia 2010 second section, dynamic turbidimetry was used for the interference test of Dexrabeprazole Sodium for Injection.Results:0.06 mg· mL-1 Dexrabeprazole Sodium had no interference and the recovery rate of bacterial endotoxin was within 50%-200%.Conclusion:Dynamic turbidimetry can be used for bacterial endotox-in test of Dexrabeprazole Sodium and the limit should be set at less than 10.0 EU· mg-1.%目的:建立注射用右旋雷贝拉唑钠细菌内毒素动态浊度法检测方法。方法:按照《中国药典》2010年版二部相关附录,用动态浊度法进行干扰试验。结果:0.06 mg · mL-1右旋雷贝拉唑钠无干扰作用,内毒素回收率均在50%~200%之间。结论:本品可用动态浊度法进行细菌内毒素定量检测,建议其内毒素限值为每1 mg右旋雷贝拉唑钠中含内毒素的量应小于10.0 EU。

  5. Pyrogenic organic matter can alter microbial communication

    Science.gov (United States)

    Masiello, Caroline; Gao, Xiaodong; Cheng, Hsiao-Ying; Silberg, Jonathan

    2016-04-01

    Soil microbes communicate with each other to manage a large range of processes that occur more efficiently when microbes are able to act simultaneously. This coordination occurs through the continuous production of signaling compounds that are easily diffused into and out of cells. As the number of microbes in a localized environment increases, the internal cellular concentration of these signaling compounds increases, and when a threshold concentration is reached, gene expression shifts, leading to altered (and coordinated) microbial behaviors. Many of these coordinated behaviors have biogeochemically important outcomes. For example, methanogenesis, denitrification, biofilm formation, and the development of plant-rhizobial symbioses are all regulated by a simple class of cell-cell signaling molecules known as acyl homoserine lactones (AHLs). Pyrogenic organic matter in soils can act to disrupt microbial communication through multiple pathways. In the case of AHLs, charcoal's very high surface area can sorb these signaling compounds, preventing microbes from detecting each others' presence (Masiello et al., 2014). In addition, the lactone ring in AHLs is vulnerable to pH increases accompanying PyOM inputs, with soil pH values higher than 7-8 leading to ring opening and compound destabilization. Different microbes use different classes of signaling compounds, and not all microbial signaling compounds are pH-vulnerable. This implies that PyOM-driven pH increases may trigger differential outcomes for Gram negative bacteria vs fungi, for example. A charcoal-driven reduction in microbes' ability to detect cell-cell communication compounds may lead to a shift in the ability of microbes to participate in key steps of C and N cycling. For example, an increase in an archaeon-specific AHL has been shown to lead to a cascade of metabolic processes that eventually results in the upregulation of CH4 production (Zhang et al., 2012). Alterations in similar AHL compounds leads to

  6. 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

  7. Endotoxin-induced endothelial fibrosis is dependent on expression of transforming growth factors β1 and β2.

    Science.gov (United States)

    Echeverría, César; Montorfano, Ignacio; Tapia, Pablo; Riedel, Claudia; Cabello-Verrugio, Claudio; Simon, Felipe

    2014-09-01

    During endotoxemia-induced inflammatory disease, bacterial endotoxins circulate in the bloodstream and interact with endothelial cells (ECs), inducing dysfunction of the ECs. We previously reported that endotoxins induce the conversion of ECs into activated fibroblasts. Through endotoxin-induced endothelial fibrosis, ECs change their morphology and their protein expression pattern, thereby suppressing endothelial markers and upregulating fibrotic proteins. The most commonly used fibrotic inducers are transforming growth factor β1 (TGF-β1) and TGF-β2. However, whether TGF-β1 and TGF-β2 participate in endotoxin-induced endothelial fibrosis remains unknown. We have shown that the endotoxin-induced endothelial fibrosis process is dependent on the TGF-β receptor, ALK5, and the activation of Smad3, a protein that is activated by ALK5 activation, thus suggesting that endotoxin elicits TGF-β production to mediate endotoxin-induced endothelial fibrosis. Therefore, we investigated the dependence of endotoxin-induced endothelial fibrosis on the expression of TGF-β1 and TGF-β2. Endotoxin-treated ECs induced the expression and secretion of TGF-β1 and TGF-β2. TGF-β1 and TGF-β2 downregulation inhibited the endotoxin-induced changes in the endothelial marker VE-cadherin and in the fibrotic proteins α-SMA and fibronectin. Thus, endotoxin induces the production of TGF-β1 and TGF-β2 as a mechanism to promote endotoxin-induced endothelial fibrosis. To the best of our knowledge, this is the first report showing that endotoxin induces endothelial fibrosis via TGF-β secretion, which represents an emerging source of vascular dysfunction. These findings contribute to understanding the molecular mechanism of endotoxin-induced endothelial fibrosis, which could be useful in the treatment of inflammatory diseases.

  8. Determinants of house dust, endotoxin, and β-(1→3)-D-glucan in homes of Danish children

    NARCIS (Netherlands)

    Holst, G; Høst, A; Doekes, G; Meyer, H W; Madsen, A M; Sigsgaard, T

    2015-01-01

    Little is known about the geographic variation and determinants of bacterial endotoxin and β-(1,3)-D-glucan in Danish house dust. In a population of 317 children, we: (i) described loads and concentrations of floor dust, endotoxin, and β-(1→3)-D-glucan and (ii) their correlations and (iii) assessed

  9. 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.

  10. A comparative study of different strategies for removal of endotoxins from bacteriophage preparations.

    Science.gov (United States)

    Van Belleghem, Jonas D; Merabishvili, Maya; Vergauwen, Bjorn; Lavigne, Rob; Vaneechoutte, Mario

    2017-01-01

    Bacterial endotoxins have high immunogenicity. Phage biology studies as well as therapeutic phage applications necessitate highly purified phage particles. In this study, we compared combinations of seven different endotoxin removal strategies and validated their endotoxin removal efficacy for five different phages (i.e. four Pseudomonas aeruginosa phages and one Staphylococcus aureus phage). These purification strategies included Endotrap HD column purification and/or CsCl density centrifugation in combination with Endotrap purification, followed by organic solvent (1-octanol), detergent (Triton X-100), enzymatic inactivation of the endotoxin using alkaline phosphatase and CIM monolytic anion exchange chromatography. We show that CsCl density purification of the P. aeruginosa phages, at an initial concentration of 10(12)-10(13)pfu/ml, led to the strongest reduction of endotoxins, with an endotoxin removal efficacy of up to 99%, whereas additional purification methods did not result in a complete removal of endotoxins from the phage preparations and only yielded an additional endotoxin removal efficacy of 23 to 99%, sometimes accompanied with strong losses in phage titer.

  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. 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

    observed peaking at 2 h, corroborating the increasing evidence that ISGs respond immediately to bacterial endotoxins. A slower response was manifested by four extrahepatic acute phase proteins (APP) (SAA3, HP, LF and LCN2) reaching maximum levels at 5 h. We report an immediate induction of ISGs...... insights into the dynamic regulation of innate immune genes, as well as raising new questions regarding the importance of ISGs and extrahepatic APPs in leukocytes after systemic endotoxin challenge....

  13. Neutron induced degradation in nitrided pyrogenic field oxide MOS capacitors

    CERN Document Server

    Vaidya, S J; Shaikh, A M; Chandorkar, A N

    2002-01-01

    Neutron induced oxide charge trapping and generation of interface states in MOS capacitors with pyrogenic and nitrided pyrogenic field oxides have been studied. In order to assess the damage due to neutrons alone, it is necessary to account for the damage produced by the accompanying gamma rays from neutron radiation. This is done by measuring the intensity of gamma radiation accompanying neutrons at different neutron fluences at the irradiation position. MOS capacitor structures were subjected to neutron radiation in a swimming pool type of reactor. Other samples from the same batch were then subjected to an equivalent dose of gamma radiation from a Co sup 6 sup 0 source. The difference in the damage observed was used to characterize the damage caused by neutrons. It is observed that neutrons, though uncharged, are capable of causing ionization damage. This damage is found to be significant when the radiation is performed under biased conditions. Nitridation in different ambients is found to improve the radi...

  14. DMPD: Tolerance to pyrogens. [Dynamic Macrophage Pathway CSML Database

    Lifescience Database Archive (English)

    Full Text Available le (.png) SVG File (.svg) HTML File (.html) CSML File (.csml) Open .csml file with CIOPlayer Open .csm...856:116-31. (.png) (.svg) (.html) (.csml) Show Tolerance to pyrogens. PubmedID 9917872 Title Tolerance to py...l file with CIOPlayer - ※CIO Playerのご利用上の注意 Open .csml file with CIO Open .csml file with CIO - ※CIOのご利用上の注意 ...

  15. 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.

  16. Facile colorimetric method for simple and rapid detection of endotoxin based on counterion-mediated gold nanorods aggregation.

    Science.gov (United States)

    Wang, Yashan; Zhang, Daohong; Liu, Wei; Zhang, Xiao; Yu, Shaoxuan; Liu, Tao; Zhang, Wentao; Zhu, Wenxin; Wang, Jianlong

    2014-05-15

    Existence of endotoxin in food and injection products indicates bacterial contaminations and therefore poses threat to human health. Herein, a simple and rapid colorimetric method for the effective detection of endotoxin in food and injections based on counterion-mediated gold nanorods aggregation is first proposed. By taking advantage of the color change of unmodified gold nanorods resulted from endotoxin mediated gold nanorods aggregation, endotoxin could be detected in the concentration range of 0.01-0.6 μM. Further, we studied the performance of gold nanorods with different aspect ratios (2.7 and 3.3) in determination of endotoxin and found that gold nanorods with higher aspect ratio (AR) showed superiority in the sensing sensitivity of endotoxin. A good specificity for endotoxin, a detection limit of 0.0084 μM and recoveries ranging from 84% to 109% in spiked food and injection samples are obtained with the colorimetric method. Results demonstrate that the present method provides a novel and effective approach for on-site screening of endotoxin in common products, which is beneficial for monitoring and reducing the risk of bacterial contaminations in food and injections production.

  17. 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.

  18. SUBCHRONIC ENDOTOXIN INHALATION CAUSES CHRONIC AIRWAY DISEASE IN ENDOTOXIN-SENSITIVE BUT NOT ENDOTOXIN-RESISTANT MICE

    Science.gov (United States)

    SUBCHRONIC ENDOTOXIN INHALATION CAUSES CHRONIC AIRWAY DISEASE IN ENDOTOXIN-SENSITIVE BUT NOT ENDOTOXIN-RESISTANT MICE. D. M. Brass, J. D. Savov, *S. H. Gavett, ?C. George, D. A. Schwartz. Duke Univ Medical Center Durham, NC, *U.S. E.P.A. Research Triangle Park, NC, ?Univ of Iowa,...

  19. Biological Activity of Masked Endotoxin

    Science.gov (United States)

    Schwarz, Harald; Gornicec, Jan; Neuper, Theresa; Parigiani, Maria Alejandra; Wallner, Michael; Duschl, Albert; Horejs-Hoeck, Jutta

    2017-01-01

    Low endotoxin recovery (LER) is a recently discovered phenomenon describing the inability of limulus amebocyte lysate (LAL)-based assays to detect lipopolysaccharide (LPS) because of a “masking effect” caused by chelators or detergents commonly used in buffer formulations for medical products and recombinant proteins. This study investigates the masking capacities of different buffer formulations and whether masked endotoxin is biologically active. We show that both naturally occurring endotoxin as well as control standard endotoxin can be affected by LER. Furthermore, whereas masked endotoxin cannot be detected in Factor C based assays, it is still detectable in a cell-based TLR4-NF-κB-luciferase reporter gene assay. Moreover, in primary human monocytes, masked LPS induces the expression of pro-inflammatory cytokines and surface activation markers even at very low concentrations. We therefore conclude that masked LPS is a potent trigger of immune responses, which emphasizes the potential danger of masked LPS, as it may pose a health threat in pharmaceutical products or compromise experimental results. PMID:28317862

  20. 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...

  1. Endotoxin Studies And Biosolids Stabilization Research

    Science.gov (United States)

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

  2. Endotoxin: From database to measurement strategy

    NARCIS (Netherlands)

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

    2007-01-01

    Endotoxin is a well-known toxin which has been associated with several health effects. Many factors influence airborne endotoxin exposure and can cause high variability in exposure between and within workers. Additionally, since the source of endotoxin exposure - gram-negative bacteria - grow and am

  3. Soil and plant responses to pyrogenic organic matter: carbon stability and symbiotic patterns

    NARCIS (Netherlands)

    Sagrilo, E.

    2014-01-01

    Soil and plant responses to pyrogenic organic matter: carbon stability and symbiotic patterns Edvaldo Sagrilo Summary Pyrogenic organic matter (PyOM), also known as biochar, is the product of biomass combustion under low oxygen concentration. There

  4. [Endotoxin adsortion as adjuvant therapy in gram negative severe sepsis].

    Science.gov (United States)

    Candel, F J; Martínez-Sagasti, F; Borges, M; Maseda, E; Herrera-Gutiérrez, M; Garnacho-Montero, J; Maynar, F J; Zaragoza, R; Mensa, J; Azanza, J R

    2010-09-01

    The mortality rate of severe sepsis and septic shock remains still high. Within the last years a better knowledge of its physiopathology and the implementation of a group of measures addressed to a fast identification and early treatment of the septic patients have proved to reduce mortality rate. Likewise, it continues being investigated in modulating the inflammatory response and limiting the harmful action of the bacterial products on the immune system. As a result of this research some endotoxin adsorber devices have been designed to control one of the most important targets that start the inflammatory cascade when gram negative microorganisms are involved.The usefulness that these endotoxin removal devices might have as adjuvant treatment in the Septic Syndrome and its applicability are reviewed in this paper. Likewise a profile of patient that might be to the benefit of this therapy is suggested according to the current knowledge.

  5. 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。

  6. 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.

  7. Discerning the biochemical stability of pyrogenic C in soils

    Science.gov (United States)

    De la Rosa, José M.; Paneque, Marina; Contreras-Bernal, Lidia; Miller, Ana Z.; Knicker, Heike

    2016-04-01

    The soil organic matter (SOM) constitutes approximately 2/3 of the global terrestrial C pool, which corresponds to estimated 4000 Pg to a depth of 3 m [1] and therefore, the dynamics of organic carbon (OC) in soils control a large part of the terrestrial C cycle. The term Pyrogenic Carbon (PyC) comprises the whole range of pyrogenic organic materials, from partly charred material through charcoal to soot produced during fire, as well as technical chars (biochars) produced by pyrolysis of biomass. The previously common assumption of PyC being inert has long been proven wrong [2]. In theory, the pyrogenic process confers these materials a longer mean residence time in the soils than their precursors, thus the application of PyC in general and particularly biochar to soil is proposed as a valid approach to establish a significant, long-term sink for atmospheric carbon dioxide in terrestrial ecosystems [3]. Nevertheless, the knowledge concerning the biochemical recalcitrance of PyOM in soils is still limited. This study combines the analysis by 13C solid state Nuclear Magnetic Resonance Spectroscopy (13C NMR), Field Emission Scanning Electron Microscopy (FESEM), analytical pyrolysis (Py-GC/MS) and CO2 emissions in incubated pots of burned and unburned soils as well as in biochar amended and un-amended soils. By using this integrated approach we achieved a more complete understanding of the stability of different forms of PyC in the soil and the chemical changes occurring during aging. Significant differences are found between the stability of PyC. They depend on the nature of the source material, surficial properties of PyC, the pyrolysis process and the soil conditions during aging. Acknowledgements: The Marie Skłodowska-Curie actions (PCIG12-GA-2012-333784-Biocharisma project and PIEF-GA-2012-328689-DECAVE project), and the Spanish Ministry of Economy and Competitiveness (MINECO) (project PCGL2012-37041) are thanked for the financial support of the present study

  8. 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.

  9. The level of endotoxins in hemodialysis water and dialysate in Lithuanian hemodialysis centers.

    Science.gov (United States)

    Skarupskienė, Inga; Bumblytė, Inga Arūnė; Tamošaitis, Donatas; Venterienė, Jūratė; Kuzminskis, Vytautas

    2010-01-01

    The composition and quality of the dialysis fluid play an important role in the modulation of dialysis-related complications. During hemodialysis, patient's blood has a contact with dialysate through a semipermeable membrane. Bacterial endotoxins can pass through the membrane pores into the patient's blood and cause a silent chronic microinflammation. The aim of this study was to determine the level of endotoxins in hemodialysis water and dialysate in Lithuanian hemodialysis centers. Dialysis water (n=50) and dialysate (n=50) were collected from 91% (n=50) of all hemodialysis centers. The presence of bacterial endotoxins was evaluated using a sensitive Limulus amebocyte lysate test, which detects intact lipopolysaccharides. The level of endotoxins was lower than 0.25 EU/mL in 43 (86%) dialysis water samples and in 46 (92%) dialysate samples, and complied with the recommendations of the European Pharmacopoeia and the European Best Practice Guidelines for pure dialysis fluid. The dialysate of 39 (78%) Lithuanian hemodialysis centers complied with the definition of an ultrapure dialysis fluid. The water and dialysate were of insufficient quality in 14% and in 8% of Lithuanian hemodialysis centers, respectively, and this could be improved by the establishment of routine investigation of endotoxins.

  10. Translocated LPS might cause endotoxin tolerance in circulating monocytes of cystic fibrosis patients.

    Directory of Open Access Journals (Sweden)

    Rosa del Campo

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

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

    Science.gov (United States)

    Del Rio-Castillo, Antonio E.; Newman, Leon; Vázquez, Ester; Kostarelos, Kostas; Wick, Peter; Fadeel, Bengt

    2016-01-01

    Nanomaterials may be contaminated with bacterial endotoxin during production and handling, which may confound toxicological testing of these materials, not least when assessing for immunotoxicity. In the present study, we evaluated the conventional Limulus amebocyte lysate (LAL) assay for endotoxin detection in graphene based material (GBM) samples, including graphene oxide (GO) and few-layered graphene (FLG). Our results showed that some GO samples interfered with various formats of the LAL assay. To overcome this problem, we developed a TNF-α expression test (TET) using primary human monocyte-derived macrophages incubated in the presence or absence of the endotoxin inhibitor, polymyxin B sulfate, and found that this assay, performed with non-cytotoxic doses of the GBM samples, enabled unequivocal detection of endotoxin with a sensitivity that is comparable to the LAL assay. FLG also triggered TNF-α production in the presence of the LPS inhibitor, pointing to an intrinsic pro-inflammatory effect. Finally, we present guidelines for the preparation of endotoxin-free GO, validated by using the TET. PMID:27880838

  12. 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.

  13. Assessment of endotoxin activity in wastewater treatment plants.

    Science.gov (United States)

    Guizani, Mokhtar; Dhahbi, Mahmoud; Funamizu, Naoyuki

    2009-07-01

    Endotoxic material, commonly associated to biological reactions, is thought to be one of the most important constituents in water. This has become a very important topic because of the common interest in microbial products governed by the possible shift to water reuse for drinking purposes. In this light, this study was conducted to provide an assessment of endotoxic activity in reclaimed wastewater. A bacterial endotoxin test (LAL test) was applied to water samples from several wastewater treatment plants (WWTP) in Sapporo, Japan keeping in view the seasonal variation. Samples were taken from several points in WWTP (influent, effluent, return sludge, advanced treatment effluent). The findings of this study indicated that wastewater shows high endotoxin activity. The value of Endotoxin (Endo) to COD ratio in the effluent is usually higher than that of the influent. Moreover, it is found that wastewater contains initially endotoxic active material. Some of those chemicals are biodegradable and but most of them are non-biodegradable. Batch scale activated sludge studies were undertaken to understand the origin of endotoxic active material in the effluent. This study showed that those chemicals are mainly produced during biological reactions, more precisely during decay process. Moreover, raw wastewater (RWW) contains high amounts of organic matter having endotoxicity which remains in the effluent.

  14. The Limulus Amebocyte Lysate assay may be unsuitable for detecting endotoxin in blood of healthy female subjects.

    Science.gov (United States)

    Gnauck, Anne; Lentle, Roger G; Kruger, Marlena C

    2015-01-01

    We examined the factors that may influence the outcome of the Limulus Amebocyte Lysate (LAL) assay, when it is used for quantifying Gram-negative bacterial endotoxin, also referred to as lipopolysaccharide (LPS), in samples of human blood. We found that the method recommended by the manufacturers, based on the reaction time, was inaccurate with any type of serum samples due to the slowing of the initial phase of reaction, likely by serum proteins. We describe an alternative method that is more accurate for use with heated serum samples. Further, we found that components of fresh serum irreversibly sequester endotoxin but that this action may be largely prevented by dilution and heating, but only if this occurs prior to the addition of endotoxin. The tests also indicated that a number of types of proprietary plastic vacutainers appeared to contain significant amounts of endotoxin. However, even when appropriate blood collection containers and calculation methods were used, the levels of endotoxin in serum samples detected by LAL assay were unlikely to reflect the total quantities of endotoxin in that sample and more likely to reflect the capacity of a given serum sample to sequester endotoxin.

  15. Actinomycetal complexes in drained peat soils of the taiga zone upon pyrogenic succession

    Science.gov (United States)

    Zenova, G. M.; Glushkova, N. A.; Bannikov, M. V.; Shvarov, A. P.; Pozdnyakov, A. I.; Zvyagintsev, D. G.

    2008-04-01

    The number and diversity of actinomycetes in peat soils vary in dependence on the stage of pyrogenic succession. In the cultivated peat soil, the number of actinomycetes after fires decreases by three-four times, mainly at the expense of acidophilic and neutrophilic groups. An increase in the number of mycelial prokaryotes (at the expense of alkaliphilic forms) is seen on the fifth year of functioning of the pyrogenic peat soil. The species diversity of streptomycetes in peat soils also decreases after fires. An increase in the range of streptomycetal species at the expense of neutrophilic and alkaliphilic forms takes place on the fifth year of the pyrogenic succession. Parameters of the actinomycetal complex—the population density, species composition, and ecological features—are the criteria whose changes allow us to judge the state of peat soils in the course of their pyrogenic succession.

  16. 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 beta......, IL-6 and tumour necrosis factor (TNF)-alpha were examined, using ELISA assays, in culture supernatants of human peripheral blood leucocytes infected with influenza virus and subsequently incubated with endotoxin. Levels of TNF-alpha were increased compared to cells incubated with virus or endotoxin...... alone. Levels of IL-1 beta were also increased whereas levels of IL-6 were generally not enhanced. Cytokines appeared within 1-2 h of stimulation with virus or endotoxin and increased subsequently to reach maximum titres between 16 and 20 h post treatment. While levels of cytokine were much lower when...

  17. Phase transitions of pyrogenic silica suspensions: a comparison to model laponite.

    Science.gov (United States)

    Kätzel, Uwe; Richter, Thomas; Stintz, Michael; Barthel, Herbert; Gottschalk-Gaudig, Torsten

    2007-09-01

    Pyrogenic silica is often used as a thickening agent in paints, pastes, adhesives, or resins. Other applications include, e.g., abrasives in chemical mechanical planarization in the microelectronics industry. In all these applications it is essential to control the state of dispersion. Sometimes, phase transitions from the liquid to the solid state are required while in other cases they have to be completely avoided for the whole shelf life. The nature and influencing parameters of the fluid-solid transition for pyrogenic silica have not been investigated so far. Most investigations deal with the phase transitions of small clay particles such as laponite. Here, we dedicate our interest to the behavior of pyrogenic silica suspensions with varying specific surface area and ionic background concentration. To get an impression of the phase transition behavior we compare our results to model laponite suspensions. We apply dynamic light scattering measurements in the backscattering regime to minimize multiple scattering contributions from concentrated pyrogenic silica suspensions. Further on we exert a decomposition of the measured autocorrelation functions into an ergodic and nonergodic contribution. The analysis of the ergodic spectrum yields two different gelation kinetics for both systems, laponite and pyrogenic silica. For laponite these are in accordance with earlier investigations. The kinetics depend on the ionic background and the solids content of the suspensions. Additionally, we used dynamic extinction spectroscopy to follow the phase transitions of pyrogenic silica on a macroscale.

  18. Tracing pyrogenic carbon (PyC) beyond terrestrial ecosystems

    Science.gov (United States)

    Wiedemeier, Daniel B.; Eglinton, Timothy I.; Hanke, Ulrich M.; Schmidt, Michael W. I.

    2015-04-01

    Combustion-derived, pyrogenic carbon (PyC) is a persistent organic carbon fraction. Due to its aromatic and condensed nature (Wiedemeier et al., 2015), it is relatively resistant against chemical and biological degradation in the environment, leading to a comparatively slow turnover, which would support carbon sequestration. PyC is produced on large scales (hundreds of teragrams) in biomass burning events such as wildfires, and by combustion of fossil fuel in industry and traffic. PyC is an inherently terrestrial product and thus has predominantly been investigated in soils and the atmosphere. Much fewer studies are available about the subsequent transport of PyC to rivers and oceans. Recently, awareness has been rising about the mobility of PyC from terrestrial to marine systems and its fate in coastal and abyssal sediments was recognized (Mitra et al, 2014). It is therefore crucial to extend our knowledge about the PyC cycle by tracing PyC through all environmental compartments. By comparing its biogeochemical behavior and budgets to that of other forms of organic carbon, it will eventually be possible to elucidate PyC's total spatiotemporal contribution to carbon sequestration. In this study, we are using a state-of-the-art PyC molecular marker method (Wiedemeier et al., 2013, Gierga et al., 2014) to trace quantity, quality as well as 13C and 14C signature of PyC in selected major river systems around the globe (Godavari, Yellow, Danube, Fraser, Mackenzie and Yukon river). Different size fractions of particulate suspended sediment are being analyzed and compared across a north-south gradient. Previous studies suggested a distinct relationship between the age of plant-derived suspended carbon and the latitude of the river system, indicating slower cycling of plant biomarkers in higher latitudes. We discuss this pattern with respect to PyC, its isotopic signature and quality and the resulting implications for the global carbon and PyC cycle. Gierga et al., 2014

  19. 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

  20. 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.

  1. The effect of the intramammary infusion of Escherichia coli endotoxin on ovulation in lactating dairy cows.

    Science.gov (United States)

    Nugent, Amelia M; Hatler, Thomas B; Silvia, William J

    2002-11-01

    The purpose of this experiment was to determine if intramammary inflammation during the periovulatory period affects the occurrence of ovulation in lactating dairy cows. Ten lactating, cyclic, Holstein dairy cows received 2 injections of prostaglandin F2alpha at eleven-day intervals, to synchronize luteolysis. The day of the second injection was designated as day 0. Ovulation was anticipated to occur 3-5 days later (on days 3-5). Beginning at the morning milking on day 1, cows received intramammary infusions of either Escherichia coli endotoxin (10 microg; n=5) or infusion vehicle (pyrogen free Hank's balanced salt solution; n=5) into 2 quarters immediately after milking. The same quarters were infused after each milking through day 4. Venous blood samples were collected daily from day -1 through 13 for determination of progesterone to monitor luteolysis and formation of a new corpus luteum. Blood samples were also collected at 4-h intervals (days -1 to 2), then at 2-h intervals (days 2 to 5) to measure concentrations of luteinizing hormone. Ovaries were examined ultrasonographically on days -1 through 5 and on day 12 to monitor follicular growth and formation of the corpus luteum. Collectively, these observations were used to determine if and when ovulation occurred. Intramammary infusion of E. coli endotoxin induced an immediate increase in the concentration of somatic cells in milk from treated quarters. However, this treatment had no effect on the occurrence or timing of ovulation. Based on ultrasonography and concentrations of progesterone, four of five cows in each treatment group appeared to have ovulated. Preovulatory surges of LH were detected within the intensive bleeding periods for three cows in each treatment group. The magnitude of the LH surge was reduced in cows receiving endotoxin.

  2. Phosphatidylcholine reverses ethanol-induced increase in transepithelial endotoxin permeability and abolishes transepithelial leukocyte activation

    DEFF Research Database (Denmark)

    Mitscherling, K.; Volynets, V.; Parlesak, Alexandr

    2009-01-01

    BACKGROUND: 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). As impaired bile flow leads to endotoxemia and the bile component phosphatidylcholine (PC) is therapeutically active in ALD, we tested the hypothesis that conjugated primary bile salts (CPBS) and PC inhibit ethanol-enhanced transepithelial permeability of endotoxin and the subsequent...... transepithelial activation of human leukocytes. METHODS: For this purpose, we used a model in which intestinal epithelial cells (Caco-2) were basolaterally cocultivated with mononuclear leukocytes. Cells were challenged apically with endotoxin from Escherichia coli K12 and were incubated with or without...

  3. 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......). As impaired bile flow leads to endotoxemia and the bile component phosphatidylcholine (PC) is therapeutically active in ALD, we tested the hypothesis that conjugated primary bile salts (CPBS) and PC inhibit ethanol-enhanced transepithelial permeability of endotoxin and the subsequent transepithelial...... activation of human leukocytes. For this purpose, we used a model in which intestinal epithelial cells (Caco-2) were basolaterally cocultivated with mononuclear leukocytes. Cells were challenged apically with endotoxin from Escherichia coli K12 and were incubated with or without the addition of CPBS (1.5 m...

  4. 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.

  5. Detection and clinical significance of inflammatory cytokines, procalcitioninand endotoxin in neonatalsepticemia

    Institute of Scientific and Technical Information of China (English)

    Bo Zhang; Wu Zhu; Wan-Hua Xie; Bang-Zhi Zeng; Xu-Hui Zhang

    2016-01-01

    Objective:To investigate the clinical significance of detection of inflammatory cytokines, procalcitionin and endotoxin in neonatal septicemia.Methods:A total of 84 cases of neonatal septicemia in our hospital from January 2013 to January 2016 were selected as observation group, and 80 cases of neonates without infection signs of the same period were selected as control group. In the early stage of admission before medication and in recovery period, the fasting venous blood was drawn respectively in the observation group, detection of IL-6, IL-8, PCT and endotoxin. In the control group, disposable blood sampling at admission to detect the above indexes.Results:Upon admittance, the levels of IL-6, IL-8, PCT and endotoxin in observation group were significantly higher than those in the control group (P<0.05); in recovery period, the levels of IL-6, IL-8, PCT and endotoxin in observation group were significantly lower than before treatment (P<0.05).Conclusions: Through comparative analysis of the levels of IL-6, IL-8, PCT and endotoxin before treatment and recovery period as well as the control group, can better reflect the dynamic changes of neonatal septicemia from bacterial infection to recovery, and is conducive to early detection of infection, a more comprehensive reflection of the infection situation, to guide clinical drug use, deserving spreading and learning.

  6. 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.

  7. Tracing pyrogenic carbon suspended in rivers on a global scale

    Science.gov (United States)

    Wiedemeier, Daniel B.; Haghipour, Negar; McIntyre, Cameron P.; Eglinton, Timothy I.; Schmidt, Michael W. I.

    2016-04-01

    Combustion-derived, pyrogenic carbon (PyC) is a persistent organic carbon fraction. Due to its aromatic and condensed nature (Wiedemeier et al., 2015), it is relatively resistant against chemical and biological degradation in the environment, leading to a comparatively slow turnover, which would support carbon sequestration. PyC is produced on large scales (hundreds of teragrams) in biomass burning events such as wildfires, and by combustion of fossil fuel in industry and traffic. PyC is an inherently terrestrial product and thus has predominantly been investigated in soils and the atmosphere. Much fewer studies are available about the subsequent transport of PyC to rivers and oceans. Recently, awareness has been rising about the mobility of PyC from terrestrial to marine systems and its fate in coastal and abyssal sediments was recognized (Mitra et al, 2013). It is therefore crucial to extend our knowledge about the PyC cycle by tracing PyC through all environmental compartments. By comparing its biogeochemical behavior and budgets to that of other forms of organic carbon, it will eventually be possible to elucidate PyC's total spatiotemporal contribution to carbon sequestration. In this study, we use a state-of-the-art PyC molecular marker method (Wiedemeier et al., 2013, Gierga et al., 2014) to trace quantity, quality as well as 13C and 14C signature of PyC in selected major river systems around the globe (Godavari, Yellow, Danube, Fraser, Mackenzie and Yukon river). Different size fractions of particulate suspended sediment are analyzed and compared across a north-south gradient. Previous studies suggested a distinct relationship between the 14C age of plant-derived suspended carbon and the latitude of the river system, indicating slower cycling of plant biomarkers in higher latitudes. We discuss this pattern with respect to PyC, its isotopic signature and quality and the resulting implications for the global carbon and PyC cycle. References Wiedemeier, D.B. et

  8. Priming of native soil organic matter by pyrogenic organic matter

    Science.gov (United States)

    DeCiucies, Silene; Dharmakeerthi, Saman; Whitman, Thea; Woolf, Dominic; Lehmann, Johannes

    2015-04-01

    Priming, in relation to pyrogenic organic matter (PyOM), describes the change in mineralization rate of non-pyrogenic ("native") soil organic matter (nSOM) due to the addition of PyOM. Priming may be 'positive', in that the addition of pyC increases the mineralization rate of native SOM, or 'negative', in that the mineralization rate of nSOM is decreased. Reasons for increased mineralization may include: (i) co-metabolism: microbial decomposition of labile C-additions increases microbial activity, and facilitates additional decomposition of npSOC by active enzymes; (ii) stimulation: substrate additions result in lifted pH, nutrient, oxygen, or water constraints resulting in increased microbial activity. Decreased mineralization may be a result of: (i) inhibition: the opposite of stimulation whereby constraints are aggravated by substrate addition. Substrate addition may also cause inhibition by interfering with enzymes or signaling compounds; (ii) preferential substrate utilization: labile fraction of PyOM additions are preferentially used up by microbes thus causing a decrease in nSOC decomposition; (iii) sorption: organic compounds are adsorbed onto PyOM surfaces, decreasing their rate of mineralization; (iv) stabilization: formation of organo-mineral associations forms stable SOC pools. We have conducted a suite of experiments to investigate these potential interactions. In a seven year long incubation study, PyOM additions increased total OM mineralization for the first 2.5 years, was equal to control after 6.2 years, and was 3% lower after 7.1 years. Cumulative nSOM mineralization was 23% less with the PyOM additions than without, and over 60% of the added PyOM was present in the labile soil fraction after the 7.1 year incubation. Two additional incubation studies, one with and without plants, showed greater nSOM mineralization in the short term and lower nSOM mineralization over the long term. Increased nSOC mineralization due to the presence of plants was

  9. 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.

  10. 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.

  11. Endospores of B subtilis are pyrogenic and activate Mono Mac 6 cells

    DEFF Research Database (Denmark)

    Moesby, Lise; Hansen, Erik W; Christensen, Jens D;

    2003-01-01

    The monocytic cell line Mono Mac 6 is sensitive to pyrogens and interleukin-6 secretion is induced after exposure to pyrogens. The aim of this study is to examine the pyrogenic activity and the interleukin-6-inducing capacity of the Gram-positive B. subtilis bacteria, endospores and isolated cell...... in a sandwich immunoassay. B. subtilis bacteria and endospores induce interleukin-6 in a dose-dependent manner. Endospores are less potent than bacteria. Lipoteichoic acid (LTA) isolated from B. subtilis induces interleukin-6 in a dose-dependent manner, whereas muramyl dipeptide (MDP) is unable to induce...... interleukin-6. Lipopolysaccharides (LPS) dose-dependently induce interleukin-6 release, but the curve differs from that of LTA both in shape and offset. The interleukin-6 secretion induced by LPS, LTA and B. subtilis bacteria can be blocked by 73-85% by an antibody directed against CD14, whereas the antibody...

  12. Soil charcoal as long-term pyrogenic carbon storage in Amazonian seasonal forests.

    Science.gov (United States)

    Turcios, Maryory M; Jaramillo, Margarita M A; do Vale, José F; Fearnside, Philip M; Barbosa, Reinaldo Imbrozio

    2016-01-01

    Forest fires (paleo + modern) have caused charcoal particles to accumulate in the soil vertical profile in Amazonia. This forest compartment is a long-term carbon reservoir with an important role in global carbon balance. Estimates of stocks remain uncertain in forests that have not been altered by deforestation but that have been impacted by understory fires and selective logging. We estimated the stock of pyrogenic carbon derived from charcoal accumulated in the soil profile of seasonal forest fragments impacted by fire and selective logging in the northern portion of Brazilian Amazonia. Sixty-nine soil cores to 1-m depth were collected in 12 forest fragments of different sizes. Charcoal stocks averaged 3.45 ± 2.17 Mg ha(-1) (2.24 ± 1.41 Mg C ha(-1) ). Pyrogenic carbon was not directly related to the size of the forest fragments. This carbon is equivalent to 1.40% (0.25% to 4.04%) of the carbon stocked in aboveground live tree biomass in these fragments. The vertical distribution of pyrogenic carbon indicates an exponential model, where the 0-30 cm depth range has 60% of the total stored. The total area of Brazil's Amazonian seasonal forests and ecotones not altered by deforestation implies 65-286 Tg of pyrogenic carbon accumulated along the soil vertical profile. This is 1.2-2.3 times the total amount of residual pyrogenic carbon formed by biomass burning worldwide in 1 year. Our analysis suggests that the accumulated charcoal in the soil vertical profile in Amazonian forests is a substantial pyrogenic carbon pool that needs to be considered in global carbon models.

  13. A comparative study of Mono Mac 6 cells, isolated mononuclear cells and Limulus amoebocyte lysate assay in pyrogen testing

    DEFF Research Database (Denmark)

    Moesby, Lise; Jensen, S; Hansen, E W;

    1999-01-01

    Pyrogen induced secretion of interleukin 6 (IL-6) in Mono Mac 6 (MM6) cells was measured. The ability of the MM6 cell culture to detect pyrogens was compared to the Limulus amoebocyte lysate (LAL) test and isolated mononuclear cells (MNC). The detection limit of MM6 for lipopolysaccharide (LPS...

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

    Science.gov (United States)

    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 collecte...

  15. Soil pyrogenic carbon lacks long-term persistence

    Science.gov (United States)

    Lutfalla, Suzanne; Abiven, Samuel; Barré, Pierre; Wiedemeier, Daniel; Christensen, Bent; Houot, Sabine; Kätterer, Thomas; Macdonald, Andy; van Oort, Fok; Chenu, Claire

    2015-04-01

    In the context of climate change, one mitigation technique currently investigated is the use of pyrogenic organic carbon (PyOC) -which is biomass turned into charcoal- to sequester carbon in soils with the hypothesis that PyOC is persistent and will not be biodegraded (or mineralized). In this study, we use the unique opportunity offered by five long term bare fallow (LTBF) experiments across Europe (Askov in Denmark, Grignon and Versailles in France, Ultuna in Sweden and Rothamsted in the United Kingdom) to compare the dynamics of PyOC and soil organic carbon (SOC) in the same plots at the decadal time scale (from 25 to 80 years of bare fallow depending on the site). Bare fallow plots were regularly sampled throughout the bare fallow duration and these samples were carefully archived. In bare fallow plots, with negligible external carbon input and with continuing biodegradation, SOC is depleting. Using the Benzene Polycarboxylic Acid (BPCA) technique to estimate the PyOC quantity and quality in the soils at different sampling dates, we investigated if PyOC content was also decreasing and compared the rates of depletion of PyOC and SOC. We found that PyOC contents decreased rapidly in soils at all sites. The loss of PyOC between the first and the last soil sampling ranged from 19.8 to 57.3% of the initial PyOC content. Furthermore, PyOC quality exhibited a similar evolution at all sites, becoming more enriched in condensed material with time. We applied a one pool model with mono-exponential decay to our data and found an average mean residence time of native PyOC of 116 years across the different sites, with a standard deviation of 15 years, just 1.6 times longer than that of SOC. Our results show that, though having a longer residence time than total SOC, PyOC content can decrease rapidly in soils suggesting that the potential for long-term C storage in soil by PyOC amendments is less than currently anticipated. Our results therefore question the concept of

  16. Towards a global assessment of pyrogenic carbon from vegetation fires

    Science.gov (United States)

    Dittmar, Thorsten; Santín, Cristina; Doerr, Stefan; Kane, Evan; Masiello, Caroline; Ohlson, Mikael; De La Rosa, Jose Maria; Preston, Caroline

    2016-04-01

    The production of pyrogenic carbon (PyC; a continuum of organic carbon (C) ranging from partially charred biomass and charcoal to soot) is a widely acknowledged C sink, with the latest estimates indicating that ~50% of the PyC produced by vegetation fires potentially sequesters C over centuries. Nevertheless, the quantitative importance of PyC in the global C balance remains contentious, and therefore, PyC is rarely considered in global C cycle and climate studies. Here we examine the robustness of existing evidence and identify the main research gaps in the production, fluxes and fate of PyC from vegetation fires. Much of the previous work on PyC production has focused on selected components of total PyC generated in vegetation fires, likely leading to underestimates. We suggest that global PyC production could be in the range of 116-385 Tg C per year, that is ~0.2-0.6% of the annual terrestrial net primary production. According to our estimations, atmospheric emissions of soot/black C might be a smaller fraction of total PyC (<2%) than previously reported. Research on the fate of PyC in the environment has mainly focused on its degradation pathways, and its accumulation and resilience either in situ (surface soils) or in ultimate sinks (marine sediments). Off-site transport, transformation and PyC storage in intermediate pools are often overlooked, which could explain the fate of a substantial fraction of the PyC mobilized annually. Rivers carry about 25-28 Tg dissolved PyC per year into the ocean where it accumulates in dissolved form over ten-thousands of year to one of the largest PyC pool on Earth. The riverine flux of suspended (particulate) PyC is largely unconstrained to date. We propose new research directions addressing gaps in the global PyC cycle to fully understand the importance of the products of burning in global C cycle dynamics. This presentation is based largely on a recent review by the same group of authors (Santín et al., 2016, Global Change

  17. 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.

  18. Network topologies and dynamics leading to endotoxin tolerance and priming in innate immune cells.

    Directory of Open Access Journals (Sweden)

    Yan Fu

    Full Text Available The innate immune system, acting as the first line of host defense, senses and adapts to foreign challenges through complex intracellular and intercellular signaling networks. Endotoxin tolerance and priming elicited by macrophages are classic examples of the complex adaptation of innate immune cells. Upon repetitive exposures to different doses of bacterial endotoxin (lipopolysaccharide or other stimulants, macrophages show either suppressed or augmented inflammatory responses compared to a single exposure to the stimulant. Endotoxin tolerance and priming are critically involved in both immune homeostasis and the pathogenesis of diverse inflammatory diseases. However, the underlying molecular mechanisms are not well understood. By means of a computational search through the parameter space of a coarse-grained three-node network with a two-stage Metropolis sampling approach, we enumerated all the network topologies that can generate priming or tolerance. We discovered three major mechanisms for priming (pathway synergy, suppressor deactivation, activator induction and one for tolerance (inhibitor persistence. These results not only explain existing experimental observations, but also reveal intriguing test scenarios for future experimental studies to clarify mechanisms of endotoxin priming and tolerance.

  19. Network Topologies and Dynamics Leading to Endotoxin Tolerance and Priming in Innate Immune Cells

    Science.gov (United States)

    Fu, Yan; Glaros, Trevor; Zhu, Meng; Wang, Ping; Wu, Zhanghan; Tyson, John; Li, Liwu; Xing, Jianhua

    2012-01-01

    The innate immune system, acting as the first line of host defense, senses and adapts to foreign challenges through complex intracellular and intercellular signaling networks. Endotoxin tolerance and priming elicited by macrophages are classic examples of the complex adaptation of innate immune cells. Upon repetitive exposures to different doses of bacterial endotoxin (lipopolysaccharide) or other stimulants, macrophages show either suppressed or augmented inflammatory responses compared to a single exposure to the stimulant. Endotoxin tolerance and priming are critically involved in both immune homeostasis and the pathogenesis of diverse inflammatory diseases. However, the underlying molecular mechanisms are not well understood. By means of a computational search through the parameter space of a coarse-grained three-node network with a two-stage Metropolis sampling approach, we enumerated all the network topologies that can generate priming or tolerance. We discovered three major mechanisms for priming (pathway synergy, suppressor deactivation, activator induction) and one for tolerance (inhibitor persistence). These results not only explain existing experimental observations, but also reveal intriguing test scenarios for future experimental studies to clarify mechanisms of endotoxin priming and tolerance.

  20. Dry and moist heat sterilisation cannot inactivate pyrogenicity of Gram positive microorganisms

    DEFF Research Database (Denmark)

    Moesby, Lise; Hansen, Erik W; Christensen, Jens D;

    2005-01-01

    inducing capacity of S. aureus. On the contrary moist heat sterilisation at 134 degrees C for 10, 20 and 40 min significantly increases the interleukin-6 inducing capacity compared to untreated S. aureus. This is confirmed in the rabbit pyrogen test. Dry heat sterilisation of B. subtilis endospores at 220...

  1. Natural and pyrogenic humic acids at goethite and natural oxide surfaces interacting with phosphate

    NARCIS (Netherlands)

    Hiemstra, T.; Mia, S.; Duhaut, P.B.; Molleman, B.

    2013-01-01

    Fulvic and humic acids have a large variability in binding to metal (hydr) oxide surfaces and interact differently with oxyanions, as examined here experimentally. Pyrogenic humic acid has been included in our study since it will be released to the environment in the case of large-scale application

  2. Characterization and Phenanthrene Sorption of Natural and Pyrogenic Organic Matter Fractions.

    Science.gov (United States)

    Jin, Jie; Sun, Ke; Wang, Ziying; Yang, Yan; Han, Lanfang; Xing, Baoshan

    2017-03-07

    Pyrogenic humic acid (HA) is released into the environment during the large-scale application of biochar. However, the biogeochemistry of pyrogenic organic matter (PyOM) fractions and their sorption of hydrophobic organic compounds (HOCs) are poorly understood in comparison with natural organic matter (NOM) fractions. HA and humin (HM) fractions isolated from soils and the oxidized biochars were characterized. Sorption of phenanthrene (PHE) by these fractions was also examined. The characterization results demonstrate that pyrogenic HAs are different from natural HAs, with the former having lower atomic H/C ratios, more abundant aromatic C, and higher concentrations of surface carboxylic groups. Compared with the fresh biochars, the Koc of PHE on their oxidized biochars, pyrogenic HA, and HM fractions were undiminished, which is encouraging for the use of biochar in soil remediation. The PyOM fractions exhibited stronger nonlinear sorption than the NOM fractions. In addition, the PyOM fractions had higher sorption capacity than the NOM fractions due to their low polar C content and high aryl C content. The results obtained from this work will shed new light on the impact of the addition of biochar on the biogeochemistry of soil organic matter and on the fate of HOCs in biochar-amended soil.

  3. High throughput pyrogenic carbon (biochar) characterisation and quantification by liquid chromatography

    NARCIS (Netherlands)

    Cerqueira, W.V.; Rittl, T.F.; Novotny, E.H.; Pereira Netto, A.D.

    2015-01-01

    Characterisation and quantification of the carbonaceous polyaromatic structure of pyrogenic carbon (PyC) are of paramount importance to evaluate the role of PyC in soil carbon sequestration. A new method of rapid resolution liquid chromatography with UV diode array detection (RRLC-UV-DAD) was dev

  4. Phospholipid imprinted polymers as selective endotoxin scavengers

    Science.gov (United States)

    Sulc, Robert; Szekely, Gyorgy; Shinde, Sudhirkumar; Wierzbicka, Celina; Vilela, Filipe; Bauer, David; Sellergren, Börje

    2017-01-01

    Herein we explore phospholipid imprinting as a means to design receptors for complex glycolipids comprising the toxic lipopolysaccharide endotoxin. A series of polymerizable bis-imidazolium and urea hosts were evaluated as cationic and neutral hosts for phosphates and phosphonates, the latter used as mimics of the phospholipid head groups. The bis-imidazolium hosts interacted with the guests in a cooperative manner leading to the presence of tight and well defined 1:2 ternary complexes. Optimized monomer combinations were subsequently used for imprinting of phosphatidic acid as an endotoxin dummy template. Presence of the aforementioned ternary complexes during polymerization resulted in imprinting of lipid dimers – the latter believed to crudely mimic the endotoxin Lipid A motif. The polymers were characterized with respect to template rebinding, binding affinity, capacity and common structural properties, leading to the identification of polymers which were thereafter subjected to an industrially validated endotoxin removal test. Two of the polymers were capable of removing endotoxin down to levels well below the accepted threshold (0.005 EU/mg API) in pharmaceutical production. PMID:28303896

  5. Phospholipid imprinted polymers as selective endotoxin scavengers

    Science.gov (United States)

    Sulc, Robert; Szekely, Gyorgy; Shinde, Sudhirkumar; Wierzbicka, Celina; Vilela, Filipe; Bauer, David; Sellergren, Börje

    2017-03-01

    Herein we explore phospholipid imprinting as a means to design receptors for complex glycolipids comprising the toxic lipopolysaccharide endotoxin. A series of polymerizable bis-imidazolium and urea hosts were evaluated as cationic and neutral hosts for phosphates and phosphonates, the latter used as mimics of the phospholipid head groups. The bis-imidazolium hosts interacted with the guests in a cooperative manner leading to the presence of tight and well defined 1:2 ternary complexes. Optimized monomer combinations were subsequently used for imprinting of phosphatidic acid as an endotoxin dummy template. Presence of the aforementioned ternary complexes during polymerization resulted in imprinting of lipid dimers – the latter believed to crudely mimic the endotoxin Lipid A motif. The polymers were characterized with respect to template rebinding, binding affinity, capacity and common structural properties, leading to the identification of polymers which were thereafter subjected to an industrially validated endotoxin removal test. Two of the polymers were capable of removing endotoxin down to levels well below the accepted threshold (0.005 EU/mg API) in pharmaceutical production.

  6. 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.

  7. 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).

  8. 体外新鲜人血法检测热原相关因子IL-1β的条件优化及初步方法学研究%Optimization and Preliminary Methodology Study of in vitro Fresh Human Whole Blood Detection Method for Pyrogen-related Factor IL-1β

    Institute of Scientific and Technical Information of China (English)

    王文佳; 陈志明; 庞智慧; 方海顺; 何华红; 李薇

    2016-01-01

    OBJECTIVE:To establish the detection method for pyrogen-related factor interleukin 1β(IL-1β)through optimiz-ing detection condition,and to conduct preliminary methodology study. METHODS:The in vitro fresh human whole blood detec-tion method was used. The bacterial endotoxin standard solution(5,2,0.8,0.32 EU/ml)were added into diluted blood;using di-luted RPMI 1640 as blank control,the content of IL-1β in blood sample was determined by ELISA after incubation. The relation-ship of the addition of different attenuants(RPMI 1640 culture,sterilized normal saline)and fetal bovine serum,final dilution vol-ume fraction(40%,20%,10%and 8.3%)and storage duration(2,5,6,8,26 h)with the contents of endotoxin IL-1βwere in-vestigated,and related coefficient and detection limits were calculated. Different dilution times of Qingkailing injection and Ginaton injection samples and interference solutions were added into diluted blood to detect their recovery. RESULTS:The results indicated that RPMI 1640 media and 40% diluted blood was more sensitive(detection limit was 0.128 EU/ml,r=0.993);while the addition of fetal bovine serum didn’t influence the results. The detection limits of blood sample storied at 4 ℃ for 26 h were 0.128 EU/ml (r>0.990). When Qingkailing injection and Ginaton injection were diluted 10,32 and more times,the detection method was not interfered and the recovery ranged 68%-118%. CONCLUSIONS:Established in vitro fresh human whole blood detection method can be used for the detection of pyrogen,and provides trial evidence for the pyrogen detection of TCM injection.%目的:建立热原相关因子白细胞介素1β(IL-1β)的检测方法并优化其条件,同时进行初步方法学研究。方法:采用体外新鲜人血法。将5、2、0.8、0.32 EU/ml的细菌内毒素标准溶液加入稀释血液中,同时用稀释后的RPMI 1640作为空白对照,培养后采用酶联免疫吸附(ELISA)试验测定血液中释放的IL-1β含量。

  9. 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.

  10. 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.

  11. 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.

  12. 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

    (Balasubramanian et al. 2012). Moisture has great influence on fungal growth and viability of bacteria. Likewise the temperature, ventilation rate and construction details reflected in the year of construction influence the moisture level in dwellings (Hägerhed-Engman, 2008; Ren et al. 2001; Bornehag et al. 2004...... 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...... envelope (Frankel et al. 2012). Bacteria and bacterial endotoxins may enter buildings through the same pathways as fungi. However, a study of Madsen et al. (2012) has shown that concentrations of bacteria in indoor settled dust are significantly affected by presence of occupants. Endotoxins are present...

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

    NARCIS (Netherlands)

    Matson, K.D.; Versteegh, M.A.; Velde, van der M.; Tieleman, B.I.

    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

  14. The impact of pyrogenic C on soil functioning : a study using ancient killn soil as a model system

    Science.gov (United States)

    Rumpel, Cornelia; Naisse, Christophe; Thi Ngo, Phuong; Davasse, Bernard; Girardin, Cyril; Chabbi, Abad

    2016-04-01

    The long-term effect of pyrogenic C on the physicochemical and biological functioning of soils is poorly understood. We used ancient killn soils as model systems in order to investigate soil properties after four centuries of pyrogenic C addition. In particular we were interested in the effect of the pyrogenic C amendment on the (micro-)biological functioning of the soil. We analysed for physicochemical properties, C mineralisation as well as C dynamics following input of 13C labelled charcoal and plant residues. Our results show compared to soil without any addition, that pyrogenic C amendment led in the long term to more rapid decomposition of the new materials. The decomposition rate was increased by about 17%. In contrast,a negative priming effect reduced soil organic carbon mineralization by about 30%. Soil physicochemical poperties, i.e. clay content, cation exchange and nutrient availability were durably improved in soil amended with pyrogenic C four centuries ago. These changes probably promoted higher microbial activity and thus intense mineralization when new plant litter was added. On the contrary, charcoal was degraded at a similar rate compared to soil without pyrogenic C amendment. Thus no specific adaptation of microorganism to charcoal degradation was observed even after several centuries. The negative priming effect induced by charcoal additiion can be due to a physical protection of the soluble carbon fraction at the surfaces of new charcoal. In contrast, the negative priming effect induced by plant residue input may be more likely due to a shift of substrate utilisation by microbial communities evolving in a nutrient-rich environment. Our results demonstrate that pyrogenic C addition modifies the carbon dynamic of soils in the long-term. We propose a conceptual model accounting for the alterations of soil functioning in the long term after pyrogenic C addition.

  15. Toxoids of Streptococcal Pyrogenic Exotoxin A Are Protective in Rabbit Models of Streptococcal Toxic Shock Syndrome

    OpenAIRE

    Roggiani, Manuela; Stoehr, Jennifer A.; Olmsted, Stephen B.; Matsuka, Yury V.; Pillai, Subramonia; Ohlendorf, Douglas H.; Schlievert, Patrick M.

    2000-01-01

    Streptococcal pyrogenic exotoxins (SPEs) are superantigens that have been implicated in causing streptococcal toxic shock syndrome (STSS). Most notably, SPE serotype A is made by nearly all M-protein serotype 1 and 3 streptococci, the M types most associated with the illness (these strains contain one or more other SPEs, and those proteins are likely also to contribute to disease). We have prepared double-, triple-, and hexa-amino-acid mutants of SPE A by PCR and other mutagenesis procedures....

  16. Detecting endotoxin with a flow cytometry-based magnetic aptasensor.

    Science.gov (United States)

    Zuo, Ming-Yan; Chen, Li-Juan; Jiang, Hao; Tan, Lin; Luo, Zhao-Feng; Wang, Yan-Mei

    2014-12-01

    Endotoxin, which is also known as lipopolysaccharide (LPS), is a marker for intruding gram-negative pathogens. It is essential to detect endotoxin quickly and sensitively in a complex milieu. A new flow cytometry (FCM)-based magnetic aptasensor assay that employs two endotoxin-binding aptamers and magnetic beads has been developed to detect endotoxin. The endotoxin-conjugated sandwich complex on magnetic beads was observed by scanning confocal laser microscopy. The resulting magnetic aptasensor rapidly detected (endotoxin within a broad dynamic detection range of 10(-8) to 10(0)mg/ml in the presence of bovine serum albumin (BSA), RNA, sucrose, and glucose, which are most likely to coexist with endotoxin in the majority of biological liquids. Only 2 μl of magnetic aptasensor was required to quantify the endotoxin solution. Furthermore, the magnetic aptasensor could be regenerated seven times and still presented an outstanding response to the endotoxin solution. Therefore, the magnetic aptasensor exhibited high sensitivity, selectivity, and reproducibility, thereby serving as a powerful tool for the quality control and high-throughput detection of endotoxin in the food and pharmaceutical industries.

  17. Airborne endotoxin in fine particulate matter in Beijing

    Science.gov (United States)

    Guan, Tianjia; Yao, Maosheng; Wang, Junxia; Fang, Yanhua; Hu, Songhe; Wang, Yan; Dutta, Anindita; Yang, Junnan; Wu, Yusheng; Hu, Min; Zhu, Tong

    2014-11-01

    Endotoxin is an important biological component of particulate matter (PM) which, upon inhalation, can induce adverse health effects, and also possibly complicate the diseases in combination with other pollutants. From 1 March 2012 to 27 February 2013 we collected air samples using quartz filters daily for the quantification of airborne endotoxin and also fine PM (PM2.5) in Beijing, China. The geometric means for endotoxin concentration and the fraction of endotoxin in PM were 0.65 EU/m3 (range: 0.10-75.02) and 10.25 EU/mg PM2.5 (range: 0.38-1627.29), respectively. The endotoxin concentrations were shown to vary greatly with seasons, typically with high values in the spring and winter seasons. Temperature and relative humidity, as well as concentrations of sulfur dioxide and nitrogen oxides were found to be significantly correlated with airborne endotoxin concentrations (p endotoxin concentrations and natural sources of Na+, K+, Mg2+, and F-, while negative correlations were observed between endotoxin concentrations and anthropogenic sources of P, Co, Zn, As, and Tl. Oxidative potential analysis revealed that endotoxin concentrations were positively correlated with reactive oxygen species (ROS), but not dithiothreitol (DTT) of PM. This study provided the first continuous time series of airborne endotoxin concentrations in Beijing, and identifies its potential associations with atmospheric factors. The information developed here can assist in the assessment of health effects of air pollution in Beijing.

  18. 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.

  19. Surface endotoxin contamination and hemocompatibility evaluation of materials.

    Science.gov (United States)

    Maitz, Manfred F; Teichmann, Juliane; Sperling, Claudia; Werner, Carsten

    2009-07-01

    To evaluate the blood compatibility of new materials, a clear distinction between properties of the materials and effects due to surface contamination by adsorbed endotoxins is essential. This study compares direct contact approaches and elution methods with water, organic solvents, nonionic, and zwitterionic detergents for determination of surface-adsorbed endotoxin by the limulus amoebocyte lysate (LAL) test and determines the blood compatibility of various surfaces with controlled endotoxin contamination in vitro. The LAL test in direct contact with an endotoxin-contaminated surface was concluded to be not practicable for most devices and its sensitivity showed a high dependence on surface characteristics. Among the elution methods, 0.2% Tween-20 showed most stable elution characteristics and appears therefore preferable. Biological reactions at in vitro blood exposure were found to be only minimally influenced by adsorbed endotoxin during the time window of 2 h, allowing for a straightforward discrimination between materials and endotoxin-dependent reactions.

  20. ROLE OF ENDOGENOUS CARBON MONOXIDE IN ENDOTOXIN SHOCK

    Institute of Scientific and Technical Information of China (English)

    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 heme-oxygenase (HO), in endotoxin shock and the efficacy of hemin,an inducer of HO were investigated.Results. The plasma CO levels were found to be significantly increased during the course 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 animals given endotoxin.Conclusion.CO is a newly found endogenously produced mediator which may play an important role in the pathogenesis of endotoxin shock.

  1. 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.

  2. 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.

  3. Fluorescent nanodiamonds as highly stable biomarker for endotoxin verification

    Science.gov (United States)

    Bergmann, Thorsten; Burg, Jan Michael; Lilholt, Maria; Maeder, Ulf; Beer, Sebastian; Salzig, Denise; Ebrahimi, Mehrdad; Czermak, Peter; Fiebich, Martin

    2012-03-01

    Fluorescent nanodiamonds (ND) provide advantageous properties as a fluorescent biomarker for in vitro and in vivo studies. The maximum fluorescence occurs around 700 nm, they do not show photobleaching or blinking and seem to be nontoxic. After a pretreatment with strong acid fluorescent ND can be functionalized and coupled to endotoxin. Endotoxin is a decay product of bacteria and causes strong immune reactions. Therefore endotoxin has to be removed for most applications. An effective removal procedure is membrane filtration. The endotoxin, coupled to fluorescent ND can be visualized by using confocal microscopy which allows the investigation of the separation mechanisms of the filtration process within the membranes.

  4. Genetic engineering approach to develop next-generation reagents for endotoxin quantification

    Science.gov (United States)

    Ogura, Norihiko; Aketagawa, Jun; Aizawa, Maki; Kobayashi, Yuki; Kawabata, Shun-ichiro; Oda, Toshio

    2016-01-01

    The bacterial endotoxin test, which uses amebocyte lysate reagents of horseshoe crab origin, is a sensitive, reproducible and simple assay to measure endotoxin concentration. To develop sustainable raw materials for lysate reagents that do not require horseshoe crabs, three recombinant protease zymogens (factor C, derived from mammalian cells; factor B; and the proclotting enzyme derived from insect cells) were prepared using a genetic engineering technique. Recombinant cascade reagents (RCRs) were then prepared to reconstruct the reaction cascade in the amebocyte lysate reagent. The protease activity of the RCR containing recombinant factor C was much greater than that of recombinant factor C alone, indicating the efficiency of signal amplification in the cascade. Compared with the RCR containing the insect cell-derived factor C, those containing mammalian cell-derived factor C, which features different glycosylation patterns, were less susceptible to interference by the injectable drug components. The standard curve of the RCR containing mammalian cell-derived recombinant factor C had a steeper slope than the curves for those containing natural lysate reagents, suggesting a greater sensitivity to endotoxin. The present study supports the future production of recombinant reagents that do not require the use of natural resources. PMID:27913792

  5. 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

  6. 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.

  7. 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

    Little is known about the geographic variation and determinants of bacterial endotoxin and β -(1,3)-d-glucan in Danish house dust. In a population of 317 children, we: (i) described loads and concentrations of floor dust, endotoxin, and β-(1→3)-d-glucan and (ii) their correlations and (iii......) 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...... potential determinants. We found: geometric means (geometric standard deviations) 186 mg/m(2) (4.3) for dust; 5.46 × 10(3) EU/m(2) (8.0) and 31.1 × 10(3) EU/g (2.6) for endotoxin; and 142 μg/m(2) (14.3) and 0.71 × 10(3) μg/g (7.3) for β-(1→3)-d-glucan. High correlations (r > 0.75) were found between floor...

  8. High Molecular Weight Petrogenic and Pyrogenic Hydrocarbons in Aquatic Environments

    Science.gov (United States)

    Abrajano, T. A., Jr.; Yan, B.; O'Malley, V.

    2003-12-01

    compounds were introduced to shallow environments through forest fires and natural coking of crude oil ( Ballentine et al., 1996; O'Malley et al., 1997). The full development of natural microbial enzymatic systems that can utilize HMW hydrocarbons as carbon or energy source attests to the antiquity of hydrocarbon dispersal processes in the environment. The environmental concern is, therefore, primarily due to the rate and spatial scale by which petroleum products are released in modern times, particularly with respect to the environmental sensitivity of some ecosystems to these releases ( Schwarzenbach et al., 1993; Eganhouse, 1997; NRC, 2002).Crude oil is produced by diagenetic and thermal maturation of terrestrial and marine plant and animal materials in source rocks and petroleum reservoirs. Most of the petroleum in use today is produced by thermal and bacterial decomposition of phytoplankton material that once lived near the surface of the world's ocean, lake, and river waters (Tissot and Welte, 1984). Terrestrially derived organic matter can be regionally significant, and is the second major contributor to the worldwide oil inventory ( Tissot and Welte, 1984; Peters and Moldowan, 1993; Engel and Macko, 1993). The existing theories hold that the organic matter present in crude oil consists of unconverted original biopolymers and new compounds polymerized by reactions promoted by time and increasing temperature in deep geologic formations. The resulting oil can migrate from source to reservoir rocks where the new geochemical conditions may again lead to further transformation of the petrogenic compounds. Any subsequent changes in reservoir conditions brought about by uplift, interaction with aqueous fluids, or even direct human intervention (e.g., drilling, water washing) likewise could alter the geochemical makeup of the petrogenic compounds. Much of our understanding of environmental sources and fate of hydrocarbon compounds in shallow environments indeed borrowed from

  9. Use of anti-pyrogenic powder to prevent spontaneous combustion of coal

    Energy Technology Data Exchange (ETDEWEB)

    Maevskaya, V.M.; Baskakov, V.I.; Boldin, V.A.

    1986-07-01

    The paper describes the generation of endogenic fires during extraction, by the shield method, of thick seams of coal prone to spontaneous combustion, outlines the disadvantages of current system of extinguishing and preventing such fires and proposes the use of anti-pyrogenic powder for this purpose. The powder, which is composed of carbanide and diammonia phosphate and is non-toxic, inexpensive and readily available, is pumped via boreholes under pressure into the coal pillar between horizons prior to its being caved in order to prevent generation of endogenic fires in the caved area of the extraction pillar by starving the fire zone of oxygen. Successful applications of this method are described.

  10. EFFECTS OF LIME (CAO) ON THE ENDOTOXIN LEVELS OF BIOSOLIDS

    Science.gov (United States)

    Lime addition is a common practice for treating biosolids in order to meet EPA 503 requirements for land application. Since this treatment kills the majority of microorganisms, will it increase the level of endotoxins present in biosolids? And, if endotoxin levels are increased, ...

  11. 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

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

    Science.gov (United States)

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

    2011-09-01

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

  13. 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...

  14. 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.

  15. Role of erosional redistribution following wildfires in determining fate of pyrogenic carbon in the soil system

    Science.gov (United States)

    Asefaw Berhe, Asmeret; Abney, Rebecca; Hockaday, William; Fogel, Marilyn; Kuhn, Tim

    2016-04-01

    Fire, erosion, and soil carbon (C) dynamics overlap in space and time. Increased rates of erosion typically follow wildfires, and fire-altered or pyrogenic C (PyC, also referred to as black carbon) is redistributed vertically within soil profiles and laterally to lower landform positions along hillslopes, changing its C sequestration trajectory. However, we currently lack sufficient understanding on how and why the interaction of fire and erosional distribution of soil materials control persistence of bulk soil organic matter (SOM) and PyC in dynamic landscapes. In this talk, we present results from wildfires that occurred in the Sierra Nevada Mountains (USA) to demonstrate how the composition (based on stable isotope composition of 13C and 15N, and NMR analysis of OM composition) and magnitude of pyrogenic carbon redistributed by soil erosion varies considerably depending on fire severity and geomorphology of the landscape. Our findings also show that PyC is preferentially transported by erosion in high severity burn slopes, compared to areas affected by low and medium severity fires. Findings of this study are critical for better integration of biogeochemical and geomorphological approaches to derive improved representation of mechanisms that regulate SOM persistence in dynamic landscapes that routinely experience more than one perturbation.

  16. Parsing pyrogenic polycyclic aromatic hydrocarbons: forensic chemistry, receptor models, and source control policy.

    Science.gov (United States)

    O'Reilly, Kirk T; Pietari, Jaana; Boehm, Paul D

    2014-04-01

    A realistic understanding of contaminant sources is required to set appropriate control policy. Forensic chemical methods can be powerful tools in source characterization and identification, but they require a multiple-lines-of-evidence approach. Atmospheric receptor models, such as the US Environmental Protection Agency (USEPA)'s chemical mass balance (CMB), are increasingly being used to evaluate sources of pyrogenic polycyclic aromatic hydrocarbons (PAHs) in sediments. This paper describes the assumptions underlying receptor models and discusses challenges in complying with these assumptions in practice. Given the variability within, and the similarity among, pyrogenic PAH source types, model outputs are sensitive to specific inputs, and parsing among some source types may not be possible. Although still useful for identifying potential sources, the technical specialist applying these methods must describe both the results and their inherent uncertainties in a way that is understandable to nontechnical policy makers. The authors present an example case study concerning an investigation of a class of parking-lot sealers as a significant source of PAHs in urban sediment. Principal component analysis is used to evaluate published CMB model inputs and outputs. Targeted analyses of 2 areas where bans have been implemented are included. The results do not support the claim that parking-lot sealers are a significant source of PAHs in urban sediments.

  17. Stimulation tests of human growth hormone secretion by insulin, lysine vasopressin, pyrogen and glucagon

    Directory of Open Access Journals (Sweden)

    Ogawa,Norio

    1974-06-01

    Full Text Available Firstly, comparisons have been made of the secretion of human growth hormone (HGH that was induced by insulin, lysine vasopressin and pyrogen injections in order to study whether these substances can be utilized as a rapid test of HGH secretion. In insulin test, a fall of the fasting blood glucose level by 28.6% or more seemed to be sufficient to provoke adequate HGH elevation, and 9.4 ng/ml or higher HGH increment was recognized as being normal, because lysine vasopressin and pyrogen produce varying degrees of side-effects and are less specific and unpredictable in the release of HGH. Secondly, the pharmacologic effects and mechanism of action of exogenous glucagon upon the HGH secretion were studied. In normal subjects after one mg sc glucagon, there was a mean peak blood glucose level of 142. 4±3.l mg/lOO ml at 30 min, HGH levels reached a mean peak level of 22. 6±4. 8 ng/ml at 150 min, and no false negative response was noted. In patients with hypopituitarism, there was no positive response in plasma HGH levels after the sc glucagon. The present study revealed that the rise and subsequent fall of blood glucose are not the sole mechanism responsible for the effct of glucagon on HGH secretion, and that the HGH secretion in response to the sc glucagon was not triggered by cathecholamine via the stimulation of the adrenal medulla.

  18. Forensic source differentiation of petrogenic, pyrogenic, and biogenic hydrocarbons in Canadian oil sands environmental samples.

    Science.gov (United States)

    Wang, Zhendi; Yang, C; Parrott, J L; Frank, R A; Yang, Z; Brown, C E; Hollebone, B P; Landriault, M; Fieldhouse, B; Liu, Y; Zhang, G; Hewitt, L M

    2014-04-30

    To facilitate monitoring efforts, a forensic chemical fingerprinting methodology has been applied to characterize and differentiate pyrogenic (combustion derived) and biogenic (organism derived) hydrocarbons from petrogenic (petroleum derived) hydrocarbons in environmental samples from the Canadian oil sands region. Between 2009 and 2012, hundreds of oil sands environmental samples including water (snowmelt water, river water, and tailings pond water) and sediments (from river beds and tailings ponds) have been analyzed. These samples were taken from sites where assessments of wild fish health, invertebrate communities, toxicology and detailed chemistry are being conducted as part of the Canada-Alberta Joint Oil Sands Monitoring Plan (JOSMP). This study describes the distribution patterns and potential sources of PAHs from these integrated JOSMP study sites, and findings will be linked to responses in laboratory bioassays and in wild organisms collected from these same sites. It was determined that hydrocarbons in Athabasca River sediments and waters were most likely from four sources: (1) petrogenic heavy oil sands bitumen; (2) biogenic compounds; (3) petrogenic hydrocarbons of other lighter fuel oils; and (4) pyrogenic PAHs. PAHs and biomarkers detected in snowmelt water samples collected near mining operations imply that these materials are derived from oil sands particulates (from open pit mines, stacks and coke piles).

  19. Natural and pyrogenic humic acids at goethite and natural oxide surfaces interacting with phosphate.

    Science.gov (United States)

    Hiemstra, Tjisse; Mia, Shamim; Duhaut, Pierre-Benoît; Molleman, Bastiaan

    2013-08-20

    Fulvic and humic acids have a large variability in binding to metal (hydr) oxide surfaces and interact differently with oxyanions, as examined here experimentally. Pyrogenic humic acid has been included in our study since it will be released to the environment in the case of large-scale application of biochar, potentially creating Darks Earths or Terra Preta soils. A surface complexation approach has been developed that aims to describe the competitive behavior of natural organic matter (NOM) in soil as well as model systems. Modeling points unexpectedly to a strong change of the molecular conformation of humic acid (HA) with a predominant adsorption in the Stern layer domain at low NOM loading. In soil, mineral oxide surfaces remain efficiently loaded by mineral-protected organic carbon (OC), equivalent with a layer thickness of ≥ ~0.5 nm that represents at least 0.1-1.0% OC, while surface-associated OC may be even three times higher. In natural systems, surface complexation modeling should account for this pervasive NOM coverage. With our charge distribution model for NOM (NOM-CD), the pH-dependent oxyanion competition of the organo-mineral oxide fraction can be described. For pyrogenic HA, a more than 10-fold increase in dissolved phosphate is predicted at long-term applications of biochar or black carbon.

  20. 一种基于人体发热机理设计的新热原检测方法研究%A novel pyrogen testing based on the mechanism of human fever reaction

    Institute of Scientific and Technical Information of China (English)

    贺庆; 谭德讲; 李冠民; 黄清泉; 高华

    2012-01-01

    (from five donors ) was used to establish a model of human body, and was incubated with standard pyrogen sample ( LPS ) and test solution respectively, then the contents of proinflammatory cytokines ( e. g. IL - 1β, IL - 6 , TNF - a ) in incubation were determined and compared to evaluate the pyrogen activity and the contaminant of samples by ELISA. Results:①This assay indicated that IL - 1β and IL - 6 were more sensitive than TNF - α in detecting pyrogens. ② The human whole blood tests ( WBT)-IL - 1β and WBT-IL -6 could detect pyrogens from gram - negative bacterial ( LPS) , gram - positive bacterial ( LTA) , and fungi ( zymosan) , which were more sensitive than rabbit method. ③Taking IL - 1β and IL - 6 as indicators, a time period of 17 - 24 h was recommended for incubation; the blood should be stored at 37 ℃ and used in 2 h. ④ The detection limit of IL - 1β and IL - 6 in fresh human whole blood to LPS was 0. 05 EU · mL-1 (IL - 1β) and 0. 1 EU · mL-1 (IL - 6) , respectively; and the IL - 1β and IL - 6 secretions had good dose - dependent relation with LPS in the range of 0. 05 -1.6 EU · mL-1 and 0. 1 - 1. 6 EU · mL -1 , respectively. ⑤ According to primary results, the novel pyrogen tests could be widely applied to biological products,e. g. Recombinant Human Interferon αlb for Injection,Recombinant Human Granulocyte Colony - stimula-ting Factor Injection, Human Immunoglobulin ( pH 4) for Intravenous Injection, Rabies Vaccine ( Vero cell) for Human Use, Freeze - dried, Chinese traditional medicine ( Shuanghuanglian Injection) and chemical drugs ( metho-trexate for injection). Conclusion:The whole blood test,which had a wide detection spectrum and was sensitive and widely applicable, should get further validation as the supplement of conventional pyrogen tests.

  1. 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.

  2. 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.

  3. The cAMP-mediated protein kinase signal transduction pathway is involved in the pyrogenic effect of CRH in rats

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    The primary action of corticotropin releasing hormone (CRH) is stimulation of the synthesis and release of adrenocorticotropic hormone and β-endorphin from the pituitary in response to stress. In addition, a number of studies indicate that CRH exerts other physiological actions within the central nervous system which are independent of the pituitary. These include increased body temperature and thermogenesis. However, the intracellular mechanism responsible for pyrogenic action of CRH is still unclear. The purpose of these studies was to determine whether or not cAMP was involved in the pyrogenic action of CRH in the rat. Intracerebroventricular (icv) microinjection of CRH (2.5 μg, 5.0 μg, 10 μg) caused increases in colonic temperature and hypothalamus cAMP level in conscious rats. The pyrogenic effects of CRH were abolished or markedly inhibited by prior injection (icv) of an adenylate cyclase inhibitor, 2,,3,-dideoxyadenosine (DDA, 30 μg) or an inhibitor of cAMP-dependent protein kinase, adenosine-3,,5,-(cyclic) monophosphorothionate (Rp-cAMPs, 15 μg). This is the first report demonstrating the pyrogenic effcet of centrally administration of CRH on the rat via the cAMP-mediated protein kinase signal transduction pathway.

  4. How the type of pyrogenic organic matter determines the SOM quality in amended soils

    Science.gov (United States)

    Merino, Agustin; Gartzia-Bengoetxea, Nahia; Morangues, Lur; Arias-Gonzalez, Ander

    2016-04-01

    Charred biomass can be used as an organic amendment and to enhance the C sink capacity of soils. There are two types of by-products containing pyrogenic OM that could be used to improve in agricultural or forestry, biochar and wood ash. Due to their different heating conditions under which it is produced (pyrolysis, combustion and different temperatures, feedstocks,..), the properties of this pyrogenic OM might be highly variable, which could affect the SOM quality and the C sink capacity of the amended soil. The purpose of this study was to assess how SOM quality is influenced by pyrogenic organic matter with different degree of carbonization. Biochar and bottom wood ash were added to two Atlantic forest soils (Pinus radiata, 12 °C, 1200 mm) with different texture, clayey loam and sandy loam. The experiment consisted in a randomized block trials, in which different doses of biochar (0, 3, 9, 18 Mh ha-1) and wood ash (0, 1.5, 4.5, and 9 Mg ha-1) were added. The Biochar applied (pH: 9.8; C: 87 %) was produced by the pyrolysis of Myscanthus sp. at 450°C in a Pyreg® pyrolysis unit. The bottom wood ash (pH: 10.6; C: 30 %) was produced by combustion in a biomass power plant. The aromatization/carbonization was lightly higher in biochar than in wood ash. This latter by-product, in addition to the black carbon, it also contained mineral ash, as well as unburnt or lightly charred plant biomass. The evolution of soil chemical and SOM properties were monitored over three years by solid state Differential Scanning Calorimetry (DSC) and 13C CPMAS NMR. These techniques were applied in bulk samples and also in fractions of different densityes. The changes in microbial activity were studied by analysis of microbial biomass C and basal respiration and soil microbial community. Three years after applications the SOM content increased lightly in the treatment receiving the highest doses of biochar and wood ash, specially in the clay loam soil. SOM in the treated soils displayed a

  5. 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.

  6. The Influence of Pyrogenic, Biogenic and Anthropogenic Emissions on Ozone Production Downwind from Boreal Forest Fires

    Science.gov (United States)

    Finch, Douglas; Palmer, Paul

    2016-04-01

    Boreal forest fires emit pollutants that can have a strong influence on downwind surface ozone concentrations, with potential implications for exceeding air quality regulations. The influence of the mixing of pyrogenic, biogenic and anthropogenic emissions on ozone is not well understood. Using the nested 0.5° latitude x 0.667° longitude GEOS-Chem chemical transport model we track biomass burning plumes in North America. We identify the changes in key chemical reactions within these plumes as well as the sensitivity of ozone to the different emission sources. We illustrate the importance of this method using a case study of a multi-day forest fire during the BORTAS aircraft campaign over eastern Canada during summer 2011. We focus on emissions from the fire on the 17th of July and follow the plume for eight days. After the initial 24 hours of pyrogenic emissions the main source of VOCs is biogenic with increasing emissions from anthropogenic sources including outflow from Quebec City and Newfoundland. Using a Lagrangian framework, we show that the ozone production efficiency (OPE) of this plume decreases steadily as it moves away from the fire but increases rapidly as the plume reaches the east coast of Canada. Using a Eulerian framework we show that ozone mixing ratios of a east coast receptor region increase by approximately 15% even though the ozone tendency of the regional air mass is negative, which we find is due to the arrival of ozone precursors in the plume. We also consider the contribution of anthropogenic outflow over Nova Scotia that originates from the eastern seaboard of the United States to the local chemistry. Using these sensitivity model runs we generate a chemical reaction narrative for the plume trajectory that helps to understand the attribution of observed ozone variations.

  7. Physicochemical changes in pyrogenic organic matter (biochar) after 15 months of field aging

    Science.gov (United States)

    Mukherjee, A.; Zimmerman, A. R.; Hamdan, R.; Cooper, W. T.

    2014-07-01

    Predicting the effects of pyrogenic organic matter (OM) addition (either natural or intentional as in the case of biochar amendment) on soil chemistry and crop yields has been hampered by a lack of understanding of how pyrogenic OM evolves in the environment over time. This work compared the physicochemical characteristics of newly made and 15-month-field-aged biochars and biochar-soil mixtures. After aging, biochars made by pyrolysis of wood and grass at 250, 400 and 650 °C exhibited 5-fold increases in cation exchange capacity (CEC), on average; appearance of anion exchange capacity (AEC); and significant decreases in pH, ash content and nanopore surface area. Cross polarization 13C nuclear magnetic resonance (NMR) analyses indicated relative increases in O-containing functional groups, including substituted aryl, carboxyl and carbonyl C, and losses of O-alkyl groups. Similar chemical trends were observed for soil-biochar mixtures, suggesting the same biochar aging processes occurred in the soil environment. However, there was evidence for a role of soil OM-microbe-biochar interaction during aging. Field aging of soil with biochar resulted in large increases in C and N content (up to 124 and 143%, respectively) and exchange capacity (up to 43%) beyond that calculated by the weighted addition of the properties of biochar and soil aged separately. These beneficial interactive effects varied with soil and biochar type. Scanning electronic microscopy (SEM) images of biochar particles aged with soil showed colonization by microbes and widespread OM coatings. Thus, sorption of both microbially produced and soil OM are likely processes that enhanced biochar aging. Thus, biochar's full beneficial effects on soil properties likely increase over time, and proper assignment of C sequestration credits to biochar users will require consideration of soil-biochar interactions.

  8. Endotoxin-Induced Structural Transformations in Liquid Crystalline Droplets

    Science.gov (United States)

    Lin, I.-Hsin; Miller, Daniel S.; Bertics, Paul J.; Murphy, Christopher J.; de Pablo, Juan J.; Abbott, Nicholas L.

    2011-06-01

    The ordering of liquid crystals (LCs) is known to be influenced by surfaces and contaminants. Here, we report that picogram per milliliter concentrations of endotoxin in water trigger ordering transitions in micrometer-size LC droplets. The ordering transitions, which occur at surface concentrations of endotoxin that are less than 10-5 Langmuir, are not due to adsorbate-induced changes in the interfacial energy of the LC. The sensitivity of the LC to endotoxin was measured to change by six orders of magnitude with the geometry of the LC (droplet versus slab), supporting the hypothesis that interactions of endotoxin with topological defects in the LC mediate the response of the droplets. The LC ordering transitions depend strongly on glycophospholipid structure and provide new designs for responsive soft matter.

  9. 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

  10. 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.

  11. Endotoxin contamination of apolipoprotein A-I: effect on macrophage proliferation--a cautionary tale.

    Science.gov (United States)

    Jin, Xueting; Xu, Qing; Champion, Keith; Kruth, Howard S

    2015-05-01

    This technical report addresses the problem of endotoxin contamination of apolipoprotein reagents. Using a bromodeoxyuridine incorporation cell proliferation assay, we observed that human plasma ApoA-I as low as 1 μg/ml resulted in a >90% inhibition in macrophage proliferation. However, not all ApoA-I from different sources showed this effect. We considered the possibility that endotoxin contamination of the apolipoproteins contributed to the differential inhibition of macrophage cell proliferation. Endotoxin alone very potently inhibited macrophage proliferation (0.1 ng/ml inhibited macrophage proliferation>90%). Measurement of endotoxin levels in the apolipoprotein products, including an analysis of free versus total endotoxin, the latter which included endotoxin that was masked due to binding to protein, suggested that free endotoxin mediated inhibition of macrophage proliferation. Despite the use of an advanced endotoxin removal procedure and agents commonly used to inhibit endotoxin action, the potency of endotoxin precluded successful elimination of endotoxin effect. Our findings show that endotoxin contamination can significantly influence apparent apolipoprotein-mediated cell effects (or effects of any other biological products), especially when these products are tested on highly endotoxin-sensitive cells, such as macrophages.

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

    Science.gov (United States)

    Lambadi, Paramesh Ramulu; Sharma, Tarun Kumar; Kumar, Piyush; Vasnani, Priyanka; Thalluri, Sitaramanjaneya Mouli; Bisht, Neha; Pathania, Ranjana; Navani, Naveen Kumar

    2015-01-01

    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 agents to control infections.

  13. Type I interferon signalling is not required for the induction of endotoxin tolerance.

    Science.gov (United States)

    Karimi, Yalda; Poznanski, Sophie M; Vahedi, Fatemeh; Chen, Branson; Chew, Marianne V; Lee, Amanda J; Ashkar, Ali A

    2017-02-08

    Endotoxin, or LPS tolerance, is an immunomodulatory mechanism that results in a significantly diminished response to secondary LPS exposure, which may serve to protect the host against endotoxic shock. Type I interferons (IFNs) are cytokines released upon LPS binding to TLR4 and have been shown to have immunomodulatory properties. Due to this regulatory function of type I IFN, we aimed to investigate the role of type I IFN signalling in LPS tolerance. Our data suggests that type I IFN does not play a role in LPS tolerance in vitro, as both wild type and IFNAR1(-/-) peritoneal macrophages showed reduced cytokine production after secondary LPS exposure. Furthermore, both wild type and IFNAR1(-/-) mice were protected from a lethal dose of LPS after receiving three small doses to induce tolerance. However, IFNAR(-/-) mice seemed to recover faster than wild type mice, suggesting type I IFN signalling plays a detrimental role in LPS-induced sepsis. Although type I IFN may have a regulatory function in microbial infections, it does not seem to play a role in endotoxin tolerance. Type I IFN involvement in bacterial infection remains complex and further studies are needed to define the regulatory function of type I IFN signalling.

  14. [Postoperative inflammatory responses to gelatin- and collagen-impregnated Dacron grafts and changes of endotoxin].

    Science.gov (United States)

    Kawashima, T; Kamisawa, O; Fuse, K; Misawa, Y; katoh, M; Hasegawa, N; Saitoh, T; Konishi, H; Ohki, S; Hasegawa, T

    1997-04-01

    Between June, 1993 and January, 1995 a gelatin-sealed knitted Dacron (Gelseal) (g-G0, n = 7) and a collagen-sealed knitted Dacron graft (Hemashield) (g-H0, n = 8) were randomly implanted to 15 patients (pts) for replacement of thoracic aorta. We evaluated postoperative inflammatory responses and levels of Toxicolor and Endospecy which are the measurements of endotoxin. Five pts in g-G0 and 3 pts in g-H0 showed that a body temperature (BT) were above 37.5 degrees C on the 7th postoperative day (POD). In three of them (2 in g-G0 and 1 in g-H0), BT above 37.5 degrees C continued until POD 14 due to the bacterial infection. The other causes of elevation of BT were pleural effusion (2), pericardial effusion (1) and unknown origin (2). Patients without evidence of infection (n = 12) were divided into 2 groups (g-G: pts with Gelseal, n = 5, g-H: pts with Hemashield, n = 7). On POD3, postoperative BT in g-H was significantly higher than in g-G. And, on POD7, BT in g-G rose up more than in g-H. However, on POD14, BT decreased to the normal range in both groups. The values of WBC in g-G were slightly higher than in g-H and they became normal after POD7 in both groups. The levels of CRP in g-H were higher than in g-G after POD3 and in both groups they were still high on POD14. In terms of endotoxin., Toxicolor was already above the normal range from POD1 and decreased to the normal range after POD14. However, the level of Endospecy kept within normal range. In conclusion, Toxicolor-reactive substance elevates by using Gelseal and Hemashield. Its substance is not endotoxin. It would not be appropriate to consider that endotoxin is an origin of fever between POD7 and 14.

  15. 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

  16. Duration of in vivo endotoxin tolerance in horses.

    Science.gov (United States)

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

    2016-05-01

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

  17. Endotoxin increases pulmonary vascular protein permeability in the dog

    Energy Technology Data Exchange (ETDEWEB)

    Welsh, C.H.; Dauber, I.M.; Weil, J.V.

    1986-10-01

    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.

  18. 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...... 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....

  19. LOW-DOSE AIRBORNE ENDOTOXIN EXPOSURE ENHANCES BRONCHIAL RESPONSIVENESS TO INHALED ALLERGEN IN ATOPIC ASTHMATICS

    Science.gov (United States)

    Endotoxin exposure has been associated with both protection against development of TH2-immune responses during childhood and exacerbation of asthma in persons who already have allergic airway inflammation.1 Occupational and experimental inhalation exposures to endotoxin have been...

  20. Recombinant factor C (rFC) assay and gas chromatography/mass spectrometry (GC/MS) analysis of endotoxin variability in four agricultural dusts.

    Science.gov (United States)

    Saito, Rena; Cranmer, Brian K; Tessari, John D; Larsson, Lennart; Mehaffy, John M; Keefe, Thomas J; Reynolds, Stephen J

    2009-10-01

    Endotoxin exposure is a significant concern in agricultural environments due to relatively high exposure levels. The goals of this study were to determine patterns of 3-hydroxy fatty acid (3-OHFA) distribution in dusts from four types of agricultural environments (dairy, cattle feedlot, grain elevator, and corn farm) and to evaluate correlations between the results of gas chromatography/mass spectrometry (GC/MS) analysis (total endotoxin) and biological recombinant factor C (rFC) assay (free bioactive endotoxin). An existing GC/MS-MS method (for house dust) was modified to reduce sample handling and optimized for small amount (rFC assay and the modified GC/EI-MS results was feedlot (0.72) > dairy (0.53) > corn farm (0.33) > grain elevator (0.11). In livestock environments, both odd- and even-numbered carbon chain length 3-OHFAs correlated with rFC assay response. The GC/EI-MS method should be especially useful for identification of specific 3-OHFAs for endotoxins from various agricultural environments and may provide useful information for evaluating the relationship between bacterial exposure and respiratory disease among agricultural workers.

  1. Puerarin ameliorates experimental alcoholic liver injury by inhibition of endotoxin gut leakage, Kupffer cell activation, and endotoxin receptors expression.

    Science.gov (United States)

    Peng, Jing-Hua; Cui, Tuan; Huang, Fu; Chen, Liang; Zhao, Yu; Xu, Lin; Xu, Li-Li; Feng, Qin; Hu, Yi-Yang

    2013-03-01

    Puerarin, an isoflavone component extracted from Kudzu (Pueraria lobata), has been demonstrated to alleviate alcohol-related disorders. Our study examined whether puerarin ameliorates chronic alcoholic liver injury through inhibition of endotoxin gut leakage, the subsequent Kupffer cell activation, and endotoxin receptors expression. Rats were provided with the Liber-DeCarli liquid diet for 8 weeks. Puerarin (90 mg/kg or 180 mg/kg daily) was orally administered from the beginning of the third week until the end of the experiment. Chronic alcohol intake caused increased serum alanine aminotransferase, aspartate aminotransferase, hepatic gamma-glutamyl transpeptidase, and triglyceride levels as well as fatty liver and neutrophil infiltration in hepatic lobules as determined by biochemical and histologic assays. A significant increase of liver tumor necrosis factor α was detected by enzyme-linked immunosorbent assay. These pathologic effects correlated with increased endotoxin level in portal vein and upregulated protein expression of hepatic CD68, lipopolysaccharide-binding protein, CD14, Toll-like receptor 2, and Toll-like receptor 4. Meanwhile, the intestinal microvilli were observed to be sparse, shortened, and irregularity in distribution under the transmission electron microscope in conjunction with the downregulated intestinal zonula occludens-1 protein expression. These hepatic pathologic changes were significantly inhibited in puerarin-treated animals as were the endotoxin levels and hepatic CD68 and endotoxin receptors. Moreover, the pathologic changes in intestinal microvillus and the decreased intestinal zonula occludens-1 were also ameliorated with puerarin treatment. These results thus demonstrate that puerarin inhibition of endotoxin gut leakage, Kupffer cell activation, and endotoxin receptors expression is involved in the alleviation of chronic alcoholic liver injury in rats.

  2. 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.

  3. The effects of dopamine on cardiogenic and endotoxin experimental shock.

    Science.gov (United States)

    Marchetti, G; Longo, T; Merlo, L; Noseda, V

    1973-01-01

    Studies on dogs show that dopamine improves cardiac performance and increases renal and mesenteric flow. This paper investigates the cardiovascular effects of dopamine on narcotized dogs. 0.2 ml. of mercury was administered into the circumflex branch of the left coronary artery of 2 groups of anesthetized dogs to induce myocardial infarction. 1-3 mg/lg of Eschrichia coli endotoxin was injected in 2 other groups of dogs to induce endotoxin shock. Dopamine was administered intravenousely in one group with cardiogenic shock and in the other group with endotoxin shock; noradrenaline was administered in the other 2 groups. Coronary resistances increased after induction of shock and declined towards normal after dopamine was injected. The effect of dopamine on mesenteric and renal resistance was not signficant. Noradrenaline's effect on cardiac performance was similar to that of dopamine, although unlike dopamine, it contributed to a significant increase of total peripheral, coronary, mesenteric, renal and femoral resistances. The effects of dopamine on the dog's hemodynamics were less evident in endotoxin than in cardiogenic shock on account of the fact that in endotoxin shock, circulatory blood volume declines to a higher extent than in cardiogenic shock, and greater blood alterations develop, mainly acidosis. In such conditions, dopamine alone will not help the hemodynamic parameters return to normal levels. Lotto et al. reports that in shocked humans with serious metabolic acidosis, dopamine is effective only when bicarbonate solutions are infused to adjust blood PH.

  4. Evolution of endotoxin contamination during production of a therapeutic serum.

    Science.gov (United States)

    Massaldi, Hugo; Morais, Victor

    2007-01-01

    A comparative bench-scale study of endotoxin contamination is presented for two common processes of immunoglobulin purification from equine plasma: ammonium sulphate fractionation of F(ab')2 fragments and caprylic acid precipitation of non-IgG proteins. To this end, both processes were carried out under normal sterile conditions, using sanitized material and equipment and optimal water quality in a clean but open environment. Stream samples, taken at different stages from each process, were analyzed for endotoxin content by the Limulus Amebocyte Lysate (LAL) test. It was found that exogenous contamination preferentially came from endotoxins already present in reagents and/or raw materials, whereas contamination from the environment was minimal. Endogenous endotoxin accumulation, concomitant with the concentration of proteins during processing, was found to be an important factor. With classic technology, blood extraction and sterilizing filtration are critical points for both processes. It is concluded that sterility is not a sufficient condition to obtain an endotoxin-free product. Only with proper sanitization of material, and by applying the caprylic acid purification process with a starting plasma below 4-5 EU/mL, would it be possible to achieve a final product within the norm.

  5. Endotoxin Treatment of Pregnant Rats Affects Sexual Behavior of the Male Offspring

    NARCIS (Netherlands)

    Wijkstra, S.; Valkhof, N.; Koolhaas, J.M.; Schuiling, G.A.

    1991-01-01

    The offspring of endotoxin-infused pregnant rats (0.2 µg endotoxin, 53.3 min, day 18 of pregnancy) did not exhibit different behavior in the Hebb-Williams-type maze test, but the males showed aberrations in the sexual behavior test. Because endotoxin did not cross the placental barrier, it was concl

  6. 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

  7. 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

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

    NARCIS (Netherlands)

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

    1984-01-01

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

  9. ACTIVATION OF PLASMA SYSTEMS AND BLOOD-CELLS BY ENDOTOXIN IN RABBITS

    NARCIS (Netherlands)

    JANSEN, NJG; VANOEVEREN, W; HOITING, BH; WILDEVUUR, CRH

    1991-01-01

    Endotoxin plays an important role in the pathogenesis of septicaemia by activation of cellular and plasmatic systems. This study was performed to investigate the effects of infusion of endotoxin in rabbits by measuring the activation of cellular and plasma systems. Endotoxin was infused at a rate of

  10. Discrimination in Degradability of Soil Pyrogenic Organic Matter Follows a Return-On-Energy-Investment Principle

    Energy Technology Data Exchange (ETDEWEB)

    Harvey, Omar R.; Myers-Pigg, Allison N.; Kuo, Li-Jung; Singh, Bhupinder Pal; Kuehn, Kevin A.; Louchouarn, Patrick

    2016-08-16

    A fundamental understanding of biodegradability is central to elucidating the role(s) of pyrogenic organic matter (PyOM) in biogeochemical cycles. Since microbial community and ecosystem dynamics are driven by net energy flows, then a quantitative assessment of energy value versus energy requirement for oxidation of PyOM should yield important insights into their biodegradability. We used bomb calorimetry, step-wise isothermal thermogravimetric analysis (isoTGA) and 5-year in-situ bidegradation data, to develop energy-biodegradability relationships for a suite of plant- and manure-derived PyOM (n = 10). The net energy value (ΔE) for PyOM was between 4.0 and 175 kJ mol-1; with manure-derived PyOM having the highest ΔE. Thermal-oxidation activation energy (Ea) requirements ranged from 51 to 125 kJ mol-1, with wood-derived PyOM having the highest Ea requirements. We propose a return-on-investment (ROI) parameter (ΔE/Ea) for differentiating short-to-medium term biodegradability of PyOM and deciphering if biodegradation will most likely proceed via co-metabolism (ROI < 1) or direct metabolism (ROI ≥ 1). The ROI-biodegradability relationship was sigmoidal with higher biodegradability associated with PyOM of higher ROI; indicating that microbes exhibit a higher preference for “high investment value” PyOM.

  11. Carbon Mineralizability Determines Interactive Effects on Mineralization of Pyrogenic Organic Matter and Soil Organic Carbon

    Energy Technology Data Exchange (ETDEWEB)

    Whitman, Thea L.; Zhu, Zihua; Lehmann, Johannes C.

    2014-10-31

    Soil organic carbon (SOC) is a critical and active pool in the global C cycle, and the addition of pyrogenic organic matter (PyOM) has been shown to change SOC cycling, increasing or decreasing mineralization rates (often referred to as priming). We adjusted the amount of easily mineralizable C in the soil, through 1-day and 6-month pre-incubations, and in PyOM made from maple wood at 350°C, through extraction. We investigated the impact of these adjustments on C mineralization interactions, excluding pH and nutrient effects and minimizing physical effects. We found short-term increases (+20-30%) in SOC mineralization with PyOM additions in the soil pre-incubated for 6 months. Over the longer term, both the 6-month and 1-day pre-incubated soils experienced net ~10% decreases in SOC mineralization with PyOM additions. This was possibly due to stabilization of SOC on PyOM surfaces, suggested by nanoscale secondary ion mass spectrometry. Additionally, the duration of pre-incubation affected priming interactions, indicating that there may be no optimal pre-incubation time for SOC mineralization studies. We show conclusively that relative mineralizability of SOC in relation to PyOM-24 C is an important determinant of the effect of PyOM additions on SOC mineralization.

  12. 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.

  13. Bacterial DNA induces the complement system activation in serum and ascitic fluid from patients with advanced cirrhosis.

    Science.gov (United States)

    Francés, Rubén; González-Navajas, José M; Zapater, Pedro; Muñoz, Carlos; Caño, Rocío; Pascual, Sonia; Márquez, Dorkas; Santana, Francia; Pérez-Mateo, Miguel; Such, José

    2007-07-01

    Translocation of intestinal bacteria to ascitic fluid is, probably, the first step in the development of spontaneous bacterial peritonitis in patients with cirrhosis. Proteins of the complement system are soluble mediators implicated in the host immune response to bacterial infections and its activation has been traditionally considered to be an endotoxin-induced phenomenon. The aim of this study was to compare the modulation of these proteins in response to the presence of bacterial DNA and/or endotoxin in patients with advanced cirrhosis and ascites in different clinical conditions. Groups I and II consisted of patients without/with bacterial DNA. Group III included patients with spontaneous bacterial peritonitis and Group IV with patients receiving norfloxacin as secondary long-term prophylaxis of spontaneous bacterial peritonitis. Serum and ascitic fluid levels of endotoxin and truncated residues of the complement system were measured by ELISA. The complement system is triggered in response to bacterial DNA, as evidenced by significantly increased levels of C3b, membrane attack complex, and C5a in patients from Groups II and III compared with patients without bacterial DNA (Group I) and those receiving norfloxacin (Group IV). Gram classification did not further differentiate the immune response between patients within groups II and III, even though endotoxin levels were, as expected, significantly higher in patients with bacterial DNA from gram-negative microorganisms. The complement protein activation observed in patients with bacterial DNA in blood and ascitic fluid is indistinguishable from that observed in patients with spontaneous bacterial peritonitis and may occur in an endotoxin-independent manner.

  14. Immunomodulatory effects of honey cannot be distinguished from endotoxin

    DEFF Research Database (Denmark)

    Timm, Michael; Bartelt, Stine; Hansen, Erik Wind

    2008-01-01

    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...... to boiling and its immunomodulatory activity could be abrogated by the addition of polymyxin B. We speculate that the observed in vitro immunomodulatory effects of honey might solely be explained by the endotoxin content in the natural honeys....

  15. Screening of Anti-endotoxin Components from Radix Isatidis

    Institute of Scientific and Technical Information of China (English)

    WANG Wenqing; FANG Jianguo; LIU Yunhai; XIE Wei

    2006-01-01

    To find a component with the strongest anti-endotoxin effect from the four components (F021 , F022, F023, and F024 ) of chloroform extract (F02) of Radix Isatidis, the protective effects of the five components on endotoxin-challenged mice and their effects on the production of TNFα and IL-6 by macrophages from the endotoxin-challenged mice were compared. It was found that the five components all had protective effects on endotoxin-challenged mice in terms of mortality. The mortalities of the mice challenged by 2.42 mg/kg endotoxin were 30 %, 50 %, 20 %, 50 % and 60 %after treatment with F02, E021 , F022, F023, and F024. The 5 components all had inhibitory effects on the production of TNFα and IL-6 by macrophages from the mice. At a concentration of LPS of 50 ng/mL, the inhibitory rates of F02 , F021 1, F022 , F023 , and F024 for the production of TNFα and IL-6were 76.54 %, 30.86 %, 78. 60 %, 36.63 %, 2.06 % and 75.85 %, 18. 45 %, 77.68 %, 24.41%, 3.64% and at the dose of 100 ng/mL, the inhibitory rates were 49.65 %, 16.86 %, 66.97 %, 13.39 %, 7.16 % and 59.78 %, 1.28 %, 61.86 %, 2.08 %, 1.44 %, respectively. It is concluded that the component F022 of Radix Isatidis has the strongest anti-endotoxin effect.

  16. Evaluation of quantification methods of occupational endotoxin exposure.

    Science.gov (United States)

    Liebers, V; Raulf-Heimsoth, M; Linsel, G; Goldscheid, N; Düser, M; Stubel, H; Brüning, Th

    2007-11-01

    Endotoxin has been identified as important component of organic-dust exposure and is suspected as main cause of work-related adverse health effects in dusty areas. Although the determination of endotoxin levels by using the Limulus amoebocyte lysate (LAL) assay is internationally accepted, reliability and variation of values measured with this test remain a point of discussion. Therefore, the purpose of the study was to determine the influence of different parameters on endotoxin activity measured in airborne samples. This study thus analyzed: (a) dust filter extraction procedures, (b) storage of samples, (c) usage of different commercially available LAL assays, and (d) results of the whole blood assay (WBA) compared to the LAL test. Using a parallel sampler, 120 filters were loaded with dust at 4 different occupational settings and extracted in 2 labs using a standardized protocol. Parameters like Tween in the extraction medium, extraction volume, centrifugation speed, and material of tubes used for extraction were tested. The LAL test and the WBA were able to determine the differences in dust load of filters obtained from the settings investigated. In addition, results varied significantly with modifications in extraction procedures. Using Tween for filter extraction mainly influenced the resulting endotoxin activity. In addition, LAL test differences according to manufacturer of LAL test, extraction volume, and whether the samples are freshly processed or frozen also resulted in significant variations in the endotoxin levels. In conclusion, a reliable assessment of exposure to endotoxin activity is only possible if standard operation procedures (SOPs) for sampling and determination are established.

  17. 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.

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

    Science.gov (United States)

    Jacobs, J H; Krop, E J M; Borras-Santos, A; Zock, J-P; Taubel, M; Hyvarinnen, A; Pekkanen, J; Doekes, G; Heederik, D J J

    2014-04-01

    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 different climates. Our aim was to characterize endotoxin levels in primary schools and evaluate associations with potential determinants. Endotoxin levels were repeatedly assessed in 23 schools in Spain (n = 7), the Netherlands (n = 10), and Finland (n = 6) using electrostatic dustfall collectors. In total, 645 measurements were taken in 237 classrooms. Endotoxin levels differed significantly between countries; Dutch schools had the highest levels, while Finnish schools showed the lowest levels. In each country, differences in endotoxin levels were observed between schools and over the sampling periods. Estimates improved after adjustment for sampling period. Factors affecting endotoxin levels in a school differed per country. In general, endotoxin levels were higher in lower grades and in classrooms with higher occupancy. School endotoxin levels may contribute significantly to total endotoxin exposure in children and teachers. As the correlation between the repeated measurements is reasonable, single endotoxin measurements form a reasonable basis for estimating annual endotoxin levels in schools.

  19. 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.

  20. 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.

  1. Characterization, Quantification and Compound-specific Isotopic Analysis of Pyrogenic Carbon Using Benzene Polycarboxylic Acids (BPCA).

    Science.gov (United States)

    Wiedemeier, Daniel B; Lang, Susan Q; Gierga, Merle; Abiven, Samuel; Bernasconi, Stefano M; Früh-Green, Gretchen L; Hajdas, Irka; Hanke, Ulrich M; Hilf, Michael D; McIntyre, Cameron P; Scheider, Maximilian P W; Smittenberg, Rienk H; Wacker, Lukas; Wiesenberg, Guido L B; Schmidt, Michael W I

    2016-05-16

    Fire-derived, pyrogenic carbon (PyC), sometimes called black carbon (BC), is the carbonaceous solid residue of biomass and fossil fuel combustion, such as char and soot. PyC is ubiquitous in the environment due to its long persistence, and its abundance might even increase with the projected increase in global wildfire activity and the continued burning of fossil fuel. PyC is also increasingly produced from the industrial pyrolysis of organic wastes, which yields charred soil amendments (biochar). Moreover, the emergence of nanotechnology may also result in the release of PyC-like compounds to the environment. It is thus a high priority to reliably detect, characterize and quantify these charred materials in order to investigate their environmental properties and to understand their role in the carbon cycle. Here, we present the benzene polycarboxylic acid (BPCA) method, which allows the simultaneous assessment of PyC's characteristics, quantity and isotopic composition ((13)C and (14)C) on a molecular level. The method is applicable to a very wide range of environmental sample materials and detects PyC over a broad range of the combustion continuum, i.e., it is sensitive to slightly charred biomass as well as high temperature chars and soot. The BPCA protocol presented here is simple to employ, highly reproducible, as well as easily extendable and modifiable to specific requirements. It thus provides a versatile tool for the investigation of PyC in various disciplines, ranging from archeology and environmental forensics to biochar and carbon cycling research.

  2. Linking Pyrogenic Organic Matter Reactivity in Soil to its Charring Temperature and Wood Source

    Science.gov (United States)

    Filley, T. R.; Gibson, C. D.; Hatton, P. J.; Dastmalchi, K.; Chatterjee, S.; Nadelhoffer, K. J.; Stark, R. E.; Bird, J.

    2014-12-01

    Understanding the link between the chemical and structural properties of pyrogenic organic matter (PyOM) and its subsequent reactivity in soil is critical to predict how future increases in forest fire frequency and intensity will affect C and N cycling. Herein, we present results from a laboratory incubation that investigated the effects of wood species and charring temperature on the decomposition of PyOM and native soil organic carbon (SOC) dynamics in a sandy soil from a northern temperate forest (University of Michigan Biological Station, Pellston, MI, USA). PyOM was produced from highly 13C/15N-labeled red maple (RM; Acer rubrum) and jack pine (JP; Pinus banksania) at 0 (native wood), 200, 300, 450 and 600 °C. PyOM amendments to soil were at 11 % total soil C. After 3 months of this ongoing incubation, 13CO2 evolution indicates that both pyrolysis temperature and species played a significant role in PyOM and native SOC mineralization. For both species, PyOM-C mineralization decreased with increasing temperature and PyOM ≥200 °C additions decreased SOC mineralization relative to controls.. In addition, PyOM-C mineralization of RM-derived PyOM was enhanced relative to JP-derived PyOM at temperatures Soils with added RM-derived PyOM exhibited significantly lower SOC mineralization at 300 and 450 °C than from JP-derived PyOM additions. These results highlighting interactive temperature and species effects are consistent with our detailed spectroscopic, elemental and isotope analysis of the PyOM samples across this pyrolysis gradient, which shows significant physicochemical changes at 300 °C for JP and between 300 and 450 °C for RM. Efforts will be made in this paper to link PyOM structural and chemical properties to the PyOM and native SOC turnover rates.

  3. Redistribution of pyrogenic carbon from hillslopes to stream corridors following a large montane wildfire

    Science.gov (United States)

    Cotrufo, M. Francesca; Boot, Claudia M.; Kampf, Stephanie; Nelson, Peter A.; Brogan, Daniel J.; Covino, Tim; Haddix, Michelle L.; MacDonald, Lee H.; Rathburn, Sarah; Ryan-Bukett, Sandra; Schmeer, Sarah; Hall, Ed

    2016-09-01

    Pyrogenic carbon (PyC) constitutes a significant fraction of organic carbon in most soils. However, PyC soil stocks are generally smaller than what is expected from estimates of PyC produced from fire and decomposition losses, implying that other processes cause PyC loss from soils. Surface erosion has been previously suggested as one such process. To address this, following a large wildfire in the Rocky Mountains (CO, USA), we tracked PyC from the litter layer and soil, through eroded, suspended, and dissolved solids to alluvial deposits along riversides. We separated deposited sediment into high- and low-density fractions to identify preferential forms of PyC transport and quantified PyC in all samples and density fractions using benzene polycarboxylic acid markers. A few months after the fire, PyC had yet to move vertically into the mineral soil and remained in the organic layer or had been transported off site by rainfall driven overland flow. During major storm events PyC was associated with suspended sediments in river water and later identified in low-density riverbank deposits. Flows from an unusually long-duration and high magnitude rainstorm either removed or buried the riverbank sediments approximately 1 year after their deposition. We conclude that PyC redistributes after wildfire in patterns that are consistent with erosion and deposition of low-density sediments. A more complete understanding of PyC dynamics requires attention to the interaction of post fire precipitation patterns and geomorphological features that control surface erosion and deposition throughout the watershed.

  4. Integrating murine gene expression studies to understand obstructive lung disease due to chronic inhaled endotoxin.

    Directory of Open Access Journals (Sweden)

    Peggy S Lai

    Full Text Available RATIONALE: Endotoxin is a near ubiquitous environmental exposure that that has been associated with both asthma and chronic obstructive pulmonary disease (COPD. These obstructive lung diseases have a complex pathophysiology, making them difficult to study comprehensively in the context of endotoxin. Genome-wide gene expression studies have been used to identify a molecular snapshot of the response to environmental exposures. Identification of differentially expressed genes shared across all published murine models of chronic inhaled endotoxin will provide insight into the biology underlying endotoxin-associated lung disease. METHODS: We identified three published murine models with gene expression profiling after repeated low-dose inhaled endotoxin. All array data from these experiments were re-analyzed, annotated consistently, and tested for shared genes found to be differentially expressed. Additional functional comparison was conducted by testing for significant enrichment of differentially expressed genes in known pathways. The importance of this gene signature in smoking-related lung disease was assessed using hierarchical clustering in an independent experiment where mice were exposed to endotoxin, smoke, and endotoxin plus smoke. RESULTS: A 101-gene signature was detected in three murine models, more than expected by chance. The three model systems exhibit additional similarity beyond shared genes when compared at the pathway level, with increasing enrichment of inflammatory pathways associated with longer duration of endotoxin exposure. Genes and pathways important in both asthma and COPD were shared across all endotoxin models. Mice exposed to endotoxin, smoke, and smoke plus endotoxin were accurately classified with the endotoxin gene signature. CONCLUSIONS: Despite the differences in laboratory, duration of exposure, and strain of mouse used in three experimental models of chronic inhaled endotoxin, surprising similarities in gene

  5. Endotoxin exposure-response in a fiberglass manufacturing facility.

    Science.gov (United States)

    Milton, D K; Wypij, D; Kriebel, D; Walters, M D; Hammond, S K; Evans, J S

    1996-01-01

    Peak expiratory flow (PEF) and workplace exposure to endotoxin, phenolic resin, and formaldehyde were measured to investigate asthma symptoms and medication use among employees in a fiberglass wool manufacturing plant. Self-recorded PEF was obtained from 37 workers, for a total of 181 days off work and 187 days at work with concurrent personal exposure monitoring. Pre- and post-shift spirometry were obtained on at least 2 days. The 8 hr time-weighted average personal exposure ranges were endotoxin; 0.4-759 ng/m3; phenolic resin, 5.7-327 micrograms/m3; and formaldehyde, 1.2-265 micrograms/m3. Amplitude percent mean peak flow was associated with years since starting regular work in the highest endotoxin exposure area, although current assignment in that area was associated with reduced amplitude--evidence for a healthy worker effect. Exposure-response was analyzed by regression of lung function change on exposure using generalized estimating equations with robust variance estimates. Endotoxin exposure above 4 ng/m3 (8 hr time-weighted average) was associated with a decline in lung function across the work shift, and with drops in lung function 16-20 hr after exposure. Phenolic resin exposure was not consistently associated with decrements, and formaldehyde was not associated with decrements in lung function.

  6. Development of endotoxin tolerance in humans in vivo.

    NARCIS (Netherlands)

    Draisma, A.; Pickkers, P.; Bouw, M.P.W.J.M.; Hoeven, J.G. van der

    2009-01-01

    OBJECTIVE: Endotoxin (lipopolysaccharide [LPS]) tolerance is characterized by a reduced responsiveness to a subsequent LPS challenge. In animal and human in vitro experiments, LPS tolerance is associated with an attenuated response of proinflammatory cytokines and an enhanced production of anti-infl

  7. Dephosphorylation of endotoxin by alkaline phosphatase in vivo

    NARCIS (Netherlands)

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

    1997-01-01

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

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

    NARCIS (Netherlands)

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

    1984-01-01

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

  9. Endotoxin exposure and atopic sensitization in adult pig farmers

    NARCIS (Netherlands)

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

    2005-01-01

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

  10. Alteco endotoxin hemoadsorption in Gram-negative septic shock patients

    Directory of Open Access Journals (Sweden)

    Hoi Ping Shum

    2014-01-01

    Full Text Available Background and Aims: Severe sepsis and septic shock are common causes of mortality and morbidity in an intensive care unit setting. Endotoxin, derived from the outer membranes of Gram-negative bacteria, is considered a major factor in the pathogenesis of sepsis. This study investigated the effect of Alteco endotoxin hemoadsorption device on Gram-negative septic shock patients. Materials and Methods: An open, controlled, prospective, randomized, single-center trial was conducted between February 2010 and June 2012. Patients with septic shock due to intra-abdominal sepsis were randomized to either conventional therapy (n = 8 or conventional therapy plus two 2-hourly sessions of Alteco endotoxin hemoadsorption (n = 7. Primary endpoint was the Sequential Organ Failure Assessment (SOFA score changes from 0 to 72 h. Secondary end points included vasopressor requirement, PaO 2 /FiO 2 ratio (PFR, length of stay (LOS, and 28-day mortality. Results: This study was terminated early as interim analysis showed a low probability of significant findings. No significant difference was noted between the two groups with respect to change in SOFA score, vasopressor score, PFR, LOS, and 28-day mortality. Side-effect was minimal. Conclusions: We could not identify any clinical benefit on the addition of Alteco endotoxin hemoadsorption to conventional therapy in patients who suffered from intra-abdominal sepsis with shock. The side effect profile of this novel device was acceptable.

  11. Vagus nerve stimulation attenuates the systemic inflammatory response to endotoxin

    Science.gov (United States)

    Borovikova, Lyudmila V.; Ivanova, Svetlana; Zhang, Minghuang; Yang, Huan; Botchkina, Galina I.; Watkins, Linda R.; Wang, Haichao; Abumrad, Naji; Eaton, John W.; Tracey, Kevin J.

    2000-05-01

    Vertebrates achieve internal homeostasis during infection or injury by balancing the activities of proinflammatory and anti-inflammatory pathways. Endotoxin (lipopolysaccharide), produced by all gram-negative bacteria, activates macrophages to release cytokines that are potentially lethal. The central nervous system regulates systemic inflammatory responses to endotoxin through humoral mechanisms. Activation of afferent vagus nerve fibres by endotoxin or cytokines stimulates hypothalamic-pituitary-adrenal anti-inflammatory responses. However, comparatively little is known about the role of efferent vagus nerve signalling in modulating inflammation. Here, we describe a previously unrecognized, parasympathetic anti-inflammatory pathway by which the brain modulates systemic inflammatory responses to endotoxin. Acetylcholine, the principle vagal neurotransmitter, significantly attenuated the release of cytokines (tumour necrosis factor (TNF), interleukin (IL)-1β, IL-6 and IL-18), but not the anti-inflammatory cytokine IL-10, in lipopolysaccharide-stimulated human macrophage cultures. Direct electrical stimulation of the peripheral vagus nerve in vivo during lethal endotoxaemia in rats inhibited TNF synthesis in liver, attenuated peak serum TNF amounts, and prevented the development of shock.

  12. Century-scale patterns and trends of global pyrogenic carbon emissions and fire influences on terrestrial carbon balance

    Science.gov (United States)

    Yang, Jia; Tian, Hanqin; Tao, Bo; Ren, Wei; Lu, Chaoqun; Pan, Shufen; Wang, Yuhang; Liu, Yongqiang

    2015-09-01

    Fires have consumed a large amount of terrestrial organic carbon and significantly influenced terrestrial ecosystems and the physical climate system over the past century. Although biomass burning has been widely investigated at a global level in recent decades via satellite observations, less work has been conducted to examine the century-scale changes in global fire regimes and fire influences on the terrestrial carbon balance. In this study, we investigated global pyrogenic carbon emissions and fire influences on the terrestrial carbon fluxes from 1901 to 2010 by using a process-based land ecosystem model. Our results show a significant declining trend in global pyrogenic carbon emissions between the early 20th century and the mid-1980s but a significant upward trend between the mid-1980s and the 2000s as a result of more frequent fires in ecosystems with high carbon storage, such as peatlands and tropical forests. Over the past 110 years, average pyrogenic carbon emissions were estimated to be 2.43 Pg C yr-1 (1 Pg = 1015 g), and global average combustion rate (defined as carbon emissions per unit area burned) was 537.85 g C m-2 burned area. Due to the impacts of fires, the net primary productivity and carbon sink of global terrestrial ecosystems were reduced by 4.14 Pg C yr-1 and 0.57 Pg C yr-1, respectively. Our study suggests that special attention should be paid to fire activities in the peatlands and tropical forests in the future. Practical management strategies, such as minimizing forest logging and reducing the rate of cropland expansion in the humid regions, are in need to reduce fire risk and mitigate fire-induced greenhouse gases emissions.

  13. Use of Pyrogenic Carbon Geosorbents to Decrease the Mobility and Bioavailability of Pharmaceuticals in the Soil-Water-Biota Continuum

    Science.gov (United States)

    Liu, Cheng-Hua; Zhang, Yingjie; Bhalsod, Gemini; Chuang, Ya-Hui; Boyd, Stephen; Teppen, Brian; Tiedje, James; Li, Hui; Zhang, Wei

    2016-04-01

    Pharmaceuticals are emerging contaminants widely detected in soil and water environments, and concerns are mounting over their potential impact on human and ecosystem health. In particular, overuse of antibiotics (an important group of pharmaceuticals) in human medicine and animal agriculture and rapid emergence of antibiotic resistant bacteria on a global scale are threatening the health of humans, animals, and the environment. We have investigated interactions of pharmaceuticals with pyrogenic carbon geosorbents (e.g., biohar and activated carbon), bacteria, and vegetable crops in order to better understand sorption, uptake, and translocation of pharmaceuticals in the soil-water-biota continuum. Sorption of antibiotics by biochars was studied to assess the effect of biochar soil amendment in reducing the transport and bioavailability of antibiotics. Pyrogenic carbonaceous materials such as biochars and activated carbon had strong sorption capacities for antibiotics, and drastically lowed the uptake of antibiotics by an Escherichia coli, therefore demonstrating soil amendment with pyrogenic carbon geosorbents as an effective remediation strategy to reduce antibiotic transport and selection pressure for antibiotic resistant bacteria. Additionally, because consuming pharmaceutical-tainted food is a direct human exposure pathway, it is critical to investigate the residue levels of pharmaceuticals in food crops grown in contaminated soils or irrigated with reclaimed water. Therefore, we have studied the uptake and accumulations of pharmaceuticals in greenhouse-grown lettuce under overhead or surface irrigations. Preliminary results indicate that pharmaceuticals of large molecular weight and low water solubility had greater concentrations in lettuce shoots under overhead irrigation than surface irrigation. Pharmaceuticals of low molecular weight and high water solubility are less clearly influenced by irrigation methods. These results implies that irrigation scheme

  14. Endotoxin depletion of recombinant protein preparations through their preferential binding to histidine tags.

    Science.gov (United States)

    Mack, Laura; Brill, Boris; Delis, Natalia; Groner, Bernd

    2014-12-01

    The presence of endotoxins in preparations of recombinantly produced therapeutic proteins poses serious problems for patients. Endotoxins can cause fever, respiratory distress syndromes, intravascular coagulation, or endotoxic shock. A number of methods have been devised to remove endotoxins from protein preparations using separation procedures based on molecular mass or charge properties. Most of the methods are limited in their endotoxin removal capacities and lack general applicability. We are describing a biotechnological approach for endotoxin removal. This strategy exploits the observation that endotoxins form micelles that expose negative charges on their surface, leading to preferential binding of endotoxins to cationic surfaces, allowing the separation from their resident protein. Endotoxins exhibit high affinity to stretches of histidines, which are widely used tools to facilitate the purification of recombinant proteins. They bind to nickel ions and are the basis for protein purification from cellular extracts by immobilized metal affinity chromatography. We show that the thrombin-mediated cleavage of two histidine tags from the purified recombinant protein and the adsorption of these histidine tags and their associated endotoxins to a nickel affinity column result in an appreciable depletion of the endotoxins in the purified protein fraction.

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

    Science.gov (United States)

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

    2014-01-01

    Simethicone emulsion is used to regulate foaming in cell culture operations in biopharmaceutical processes. It is also a potential source of endotoxin contamination. The inactivation of endotoxins in dilute simethicone emulsions was assessed as a function of time at different steam temperatures using a Limulus amebocyte lysate kinetic chromogenic technique. Endotoxin inactivation from steam-heat treatment was fit to a four-parameter double exponential decay model, which indicated that endotoxin inactivation was biphasic, consisting of fast and slow regimes. In the fast regime, temperature-related effects were dominant. Transitioning into the slow regime, the observed temperature dependence diminished, and concentration-related effects became increasingly significant. The change in the Gibbs free energy moving through the transition state indicated that a large energy barrier must be overcome for endotoxin inactivation to occur. The corresponding Arrhenius pre-exponential factor was >10(12) s(-1) suggesting that endotoxins in aqueous solution exist as aggregates. The disorder associated with the endotoxin inactivation reaction pathway was assessed via the change in entropy moving through the transition state. This quantity was positive indicating that endotoxin inactivation may result from hydrolysis of individual endotoxin molecules, which perturbs the conformation of endotoxin aggregates, thereby modulating the biological activity observed. Steam-heat treatment decreased endotoxin levels by 1-2 logarithm (log) reduction (LRV), which may be practically relevant depending on incoming raw material endotoxin levels. Antifoam efficiency and cell culture performance were negligibly impacted following steam-heat treatment. The results from this study show that steam-heat treatment is a viable endotoxin control strategy that can be implemented to support large-scale biopharmaceutical manufacturing.

  16. 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

  17. Structural Characterization and Reactivity of Pyrogenic Water-Soluble Organic Matter Derived from Biomass Combustion

    Science.gov (United States)

    Norwood, M. J.; Louchouarn, P.; Kuo, L.

    2011-12-01

    Combustion processes, whether from natural or anthropogenic origin, are major sources of particulate matter (PM), black carbon (BC), and volatile organic carbon to the atmosphere as well as soils and aquatic environments. The ubiquitous presence of biomass combustion by-products in atmospheric particles and soils could potentially lead to a large transfer of pyrogenic water-soluble organic matter (Pyr-WSOM) to the surface of watersheds and aquatic systems. In spite of this, there is a dearth of studies that have characterized the sources, and particularly the fate, of Pyr-WSOM to aquatic systems. In the present study, Pyr-WSOM was extracted from plant-derived chars (feedstocks: honey mesquite, cordgrass, and loblolly pine) produced at a range of temperatures (150-850C), and were then characterized using elemental analyses and ATR-FTIR. Low temperature (250C) Pyr-WSOM, extracted from honey mesquite and cordgrass biochars, were then incubated with aliquots of filtered water from the Trinity River (TX) for one month under dark conditions. Consistent with prior studies on combustion molecular markers such as anhydrosugars and methoxylated phenols, the total amount of dissolved organic carbon (DOC) released from biochars peaks around 200-250C and then decreases with increasing temperature of combustion. Elemental and structural analyses of biochar-derived WSOM reflect the selective solubility of certain functional groups. For example, despite the predominance of aromatic units and soot structures in biochars formed at high temperatures, such functionalities are not as predominant in their respective Pyr-WSOM. In addition, the high proportion of O-containing functionalities suggests that Pyr-WSOM may be more biodegradable than the particulate residues of biomass combustion. Indeed, low temperature Pyr-WSOM decomposed rapidly with half-lives ranging ~30 days for total DOC to 4-5 days for specific molecular markers of biomass combustion. These rapid turnover rates are in

  18. Total and pyrogenic carbon stocks in black spruce forest floors from eastern Canada

    Science.gov (United States)

    Soucemarianadin, Laure; Quideau, Sylvie; MacKenzie, M. Derek; Munson, Alison; Boiffin, Juliette; Bernard, Guy; Wasylishen, Roderick

    2016-04-01

    In boreal forests, pyrogenic carbon (PyC), a by-product of recurrent wildfires, is an important component of the global soil C pool, although precise assessment of boreal PyC stock is scarce. In this study including 14 fire sites spreading over 600 km in the Quebec province, our aim was to better estimate total C stock and PyC stock in forest floors of Eastern Canada boreal forests. We also investigated the environmental conditions controlling the stocks and characterized the composition of the various forest floor layers. We analyzed the forest floor samples that were collected from mesic black spruce sites recently affected by fire (3-5 years) using elemental analysis and solid state 13C nuclear magnetic resonance (NMR) spectroscopy. PyC content was further estimated using a molecular mixing model on the 13C NMR data. Total C stock in forest floors averaged 5.7 ± 2.9 kg C/m2 and PyC stock 0.6 ± 0.3 kg C/m2. Total C stock was under control of the position in the landscape, with a greater accumulation of organic material on northern aspects and lower slope positions. In addition, total stock was significantly higher in spruce-dominated forest floors than in stands where jack pine was dominant. The PyC stock was significantly related to the atomic H/C ratio (R2 = 0.84) of the different organic layers. 13C NMR spectroscopy revealed a large increase in aromatic carbon in the deepest forest floor layer (humified H horizon) at the organic-mineral soil interface. The majority of the PyC stock was located in this horizon and had been formed during past high severity fires rather than during the most recent fire event. Conversely, the superficial "fresh" PyC layer, produced by early-season wildfires in 2005-2007, had NMR spectra fairly similar to unburned forest floors and comparatively low PyC stocks.

  19. White-rot fungal response to fresh and photolytically-weathered pyrogenic organic matter

    Science.gov (United States)

    Gibson, C. D.; Berry, T. D.; Wang, R.; Bird, J. A.; Filley, T. R.

    2013-12-01

    Pyrogenic organic matter (PyOM or biochar) is the product of the incomplete combustion of biomass. A better understanding of the microbial-mediated degradation of PyOM is critical to assess its role in soil C sequestration and to serve as an agricultural amendment. Recent studies have shown that PyOM additions can prime native soil C but results have been inconsistent, with studies reporting no effect, an increase, or decrease in C mineralization. This study investigated the ability of saprotrophic white-rot fungus, Trametes versicolor, to decompose an unaltered 'fresh' PyOM and a photo-oxidized PyOM. In addition, we measured PyOM-induced priming effects on the mineralization of malt extract agar media (MEA). Enriched (13C) Pinus banksiana-derived PyOM, produced at 450oC under N2, was added fresh and after 4 weeks exposure to 254 nm light to MEA. Vials containing the various types of media were then monitored for CO2 evolution and oxidative enzyme activity. We found that MEA C respired was stimulated (positive priming) by photolyzed PyOM and was inhibited with fresh PyOM addition (negative priming) relative to controls. Vetryl alcohol addition, a laccase production stimulant, resulted in less activity in the presence of PyOM compared with a control, indicating PyOM may disrupt enzyme induction processes. Loss of PyOM-13CO2 was 0.2% (× 0.001) for fresh PyOM and 1.2% (×0.001) for photolyzed PyOM C during 10 weeks averaged across media treatments. While MEA C mineralization decreased after fresh PyOM addition, both oxidative (laccase and manganese peroxidase) and hydrolytic (β glucosidase) enzyme production increased with fresh PyOM in the absence of veratryl alcohol. However, there was a decrease in its presence. These results suggest that the physiological response of this common wood decay fungus to PyOM is complex and responsive to enzymatic triggers but that PyOM itself can act to promote or suppress overall litter or soil decay by fungi.

  20. The stability of Pyrogenic Organic Matter is dependent upon its wood source and charring temperature

    Science.gov (United States)

    Gibson, C. D.; Filley, T. R.; Bird, J.; Nadelhoffer, K. J.; Stark, R. E.; Hatton, P. J.

    2015-12-01

    Fire is a major mediator of carbon (C) cycling in forests and can result in the formation of pyrogenic organic matter (PyOM). The biological reactivity of PyOM is largely dependent upon the physiochemical characteristics of source material and production temperature. As a result, PyOM can persist up to centennial time scales after deposition while simultaneously enhancing or suppressing the mineralization of native soil C (NSC). To investigate the interactive effects of PyOM source and production temperature on NSC, we added 13C-enriched red maple (RM) or jack pine (JP) pyrolyzed at 200, 300, 450 or 600°C to a low C (0.5%;), near-surface soil (0-20 cm-depth) at 60% water holding capacity and 11% of native soil C and then incubated the samples in the dark at 25⁰C for 6 months. We found that PyOM mineralization rates decreased with increasing pyrolysis temperature for either species while NSC mineralization was suppressed across all treatments with the largest decrease observed in JP 300⁰C. RM mineralization rates were consistently greater (˜5 to ˜25%) than for JP residence time (MRT) of PyOM-C was significantly higher in PyOM >300°C resulting in MRT of ~300 - 550 y compared to 8-30 y in lower temperature PyOM (0-300 °C). RM exhibited significantly lower MRT in 300°C PyOM than JP corresponding to differences observed in PyOM and NSC mineralization rates. The modelled active and slow pools of PyOM-C mineralized decreased with increasing combustion temperature. JP 300°C had a 20% smaller active pool when compared to RM suggesting that for both species, 300⁰C - 450⁰C represented a thermal transition point which was most evident in jack pine. These results highlight how differences in PyOM physiochemical characteristics linked to a species thermal transformation threshold may be a predictor in determining its biological reactivity in soil.

  1. Pyrogenic carbon distribution in mineral topsoils of the northeastern United States

    Science.gov (United States)

    Jauss, Verena; Sullivan, Patrick J.; Sanderman, Jonathan; Smith, David; Lehmann, Johannes

    2017-01-01

    Due to its slow turnover rates in soil, pyrogenic carbon (PyC) is considered an important C pool and relevant to climate change processes. Therefore, the amounts of soil PyC were compared to environmental covariates over an area of 327,757 km2 in the northeastern United States in order to understand the controls on PyC distribution over large areas. Topsoil (defined as the soil A horizon, after removal of any organic horizons) samples were collected at 165 field sites in a generalised random tessellation stratified design that corresponded to approximately 1 site per 1600 km2 and PyC was estimated from diffuse reflectance mid-infrared spectroscopy measurements using a partial least-squares regression analysis in conjunction with a large database of PyC measurements based on a solid-state 13C nuclear magnetic resonance spectroscopy technique. Three spatial models were applied to the data in order to relate critical environmental covariates to the changes in spatial density of PyC over the landscape. Regional mean density estimates of PyC were 11.0 g kg− 1 (0.84 Gg km− 2) for Ordinary Kriging, 25.8 g kg− 1(12.2 Gg km− 2) for Multivariate Linear Regression, and 26.1 g kg− 1 (12.4 Gg km− 2) for Bayesian Regression Kriging. Akaike Information Criterion (AIC) indicated that the Multivariate Linear Regression model performed best (AIC = 842.6; n = 165) compared to Ordinary Kriging (AIC = 982.4) and Bayesian Regression Kriging (AIC = 979.2). Soil PyC concentrations correlated well with total soil sulphur (P < 0.001; n = 165), plant tissue lignin (P = 0.003), and drainage class (P = 0.008). This suggests the opportunity of including related environmental parameters in the spatial assessment of PyC in soils. Better estimates of the contribution of PyC to the global carbon cycle will thus also require more accurate assessments of these covariates.

  2. Assessing aromaticity and the degree of aromatic condensation of pyrogenic carbon

    Science.gov (United States)

    Wiedemeier, D. B.; Abiven, S.; Hockaday, W. C.; Keiluweit, M.; Kleber, M.; Masiello, C. A.; McBeath, A. V.; Nico, P. S.; Pyle, L. A.; Schneider, M. P.; Smernik, R. J.; Wiesenberg, G. L.; Schmidt, M. W.

    2013-12-01

    Fire-derived, pyrogenic carbon (PyC) is a persistent organic carbon fraction in soils because it is relatively resistant against chemical and biological degradation. PyC thus represents a carbon sequestration potential in the global carbon cycle and was also reported to be potentially beneficial for soil fertility. PyC is naturally added to soils during wildfires and anthropogenically in the form of biochar, after organic waste is pyrolyzed. Aromaticity and the degree of aromatic condensation are the two main quality properties of PyC that probably determine its persistence against degradation. Consequently, the two properties largely influence the PyC's carbon sequestration potential as well as the duration, during which it can provide benefits to the soil. Aromaticity and the degree of aromatic condensation of PyC should theoretically be dependent on pyrolysis conditions, such as highest heating temperature or pyrolysis time, and also depend on the feedstock. In this study, we used two different pyrolysis procedures and four different feedstocks to produce four thermosequences of 38 chars in total, with highest heating temperatures ranging from 100 - 1000° C. The chars were then analyzed with an extensive suite of seven different methods: solid state 13C nuclear magnetic resonance (13C NMR), diffuse infrared Fourier transform spectroscopy (DRIFT), synchrotron-based near-edge X-ray absorption fine structure analysis (NEXAFS), benzene polycarboxylic acid analysis (BPCA), lipid analysis, elemental analysis and helium pycnometry. These methods allowed to infer the aromaticity and the degree of aromatic condensation of the differently pyrolyzed materials. Using multivariate statistical methods, aromaticity and the degree of aromatic condensation could successfully be linked to highest heating temperature and other pyrolysis conditions because characteristic patterns of the two aromatic properties could be observed by different methods throughout all four

  3. Improved quantification of pyrogenic carbon in soils and sediments by a HPLC-DAD method

    Science.gov (United States)

    Wiedemeier, D. B.; Hilf, M. D.; Smittenberg, R. H.; Schmidt, M. W. I.

    2012-04-01

    Fire-derived (pyrogenic) carbon (PyC) is produced by the incomplete combustion of biomass, for example during wildfires. It can persist in the environment for a long time due to its relative resistance against biological and chemical breakdown. Its accurate quantification in soils and sediments is of great interest because the slow turn-over of PyC has implications for the global carbon cycle and carbon budget calculations. Moreover, PyC in pedological and sedimentological records can be used to reconstruct wildfire history or to investigate historical periods like the industrialization. A whole suite of PyC quantification methods exists because PyC is not a defined chemical structure but rather a continuum of thermally altered biomass. The benzene polycarboxylic acids (BPCA) analysis is a molecular marker method that was shown to give conservative estimates of PyC quantity in soils. In addition, it yields qualitative information about the degree of aromaticity and condensation of PyC. The commonly used BPCA method consists in digesting samples with nitric acid that breaks down the PyC into a suite of BPCAs, which are cleaned, derivatized and finally analyzed by gas chromatography-flame ionization detection (GC-FID). Here, we present a modified BPCA method for soils and sediments that uses a high performance liquid chromatography system coupled to diode array detection (HPLC-DAD). We demonstrate that this method greatly enhances the reproducibility of PyC quantification in soil and sediment samples while significantly reducing analysis time. Moreover, much less sample material is needed for precise PyC quantification and we show that the HPLC-DAD method yields consistently higher PyC contents than the GC-FID method. Additionally, the modified method also facilitates δ13C and 14C measurements of the PyC fraction in these complex matrix samples. The isotopic information further improves the assessment of PyC budgets in the environment and the reconstruction of past

  4. Cupric Oxide (CuO) Oxidation Detects Pyrogenic Carbon in Burnt Organic Matter and Soils.

    Science.gov (United States)

    Hatten, Jeff; Goñi, Miguel

    2016-01-01

    Wildfire greatly impacts the composition and quantity of organic carbon stocks within watersheds. Most methods used to measure the contributions of fire altered organic carbon-i.e. pyrogenic organic carbon (Py-OC) in natural samples are designed to quantify specific fractions such as black carbon or polyaromatic hydrocarbons. In contrast, the CuO oxidation procedure yields a variety of products derived from a variety of precursors, including both unaltered and thermally altered sources. Here, we test whether or not the benzene carboxylic acid and hydroxy benzoic acid (BCA) products obtained by CuO oxidation provide a robust indicator of Py-OC and compare them to non-Py-OC biomarkers of lignin. O and A horizons from microcosms were burned in the laboratory at varying levels of fire severity and subsequently incubated for 6 months. All soils were analyzed for total OC and N and were analyzed by CuO oxidation. All BCAs appeared to be preserved or created to some degree during burning while lignin phenols appeared to be altered or destroyed to varying extents dependent on fire severity. We found two specific CuO oxidation products, o-hydroxybenzoic acid (oBd) and 1,2,4-benzenetricarboxylic acid (BTC2) that responded strongly to burn severity and withstood degradation during post-burning microbial incubations. Interestingly, we found that benzene di- and tricarboxylic acids (BDC and BTC, respectively) were much more reactive than vanillyl phenols during the incubation as a possible result of physical protection of vanillyl phenols in the interior of char particles or CuO oxidation derived BCAs originating from biologically available classes of Py-OC. We found that the ability of these compounds to predict relative Py-OC content in burned samples improved when normalized by their respective BCA class (i.e. benzene monocarboxylic acids (BA) and BTC, respectively) and when BTC was normalized to total lignin yields (BTC:Lig). The major trends in BCAs imparted by burning

  5. 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

  6. Quantitative characterization of agglomerates and aggregates of pyrogenic and precipitated amorphous silica nanomaterials by transmission electron microscopy

    Directory of Open Access Journals (Sweden)

    De Temmerman Pieter-Jan

    2012-06-01

    Full Text Available Abstract Background The interaction of a nanomaterial (NM with a biological system depends not only on the size of its primary particles but also on the size, shape and surface topology of its aggregates and agglomerates. A method based on transmission electron microscopy (TEM, to visualize the NM and on image analysis, to measure detected features quantitatively, was assessed for its capacity to characterize the aggregates and agglomerates of precipitated and pyrogenic synthetic amorphous silicon dioxide (SAS, or silica, NM. Results Bright field (BF TEM combined with systematic random imaging and semi-automatic image analysis allows measuring the properties of SAS NM quantitatively. Automation allows measuring multiple and arithmetically complex parameters simultaneously on high numbers of detected particles. This reduces operator-induced bias and assures a statistically relevant number of measurements, avoiding the tedious repetitive task of manual measurements. Access to multiple parameters further allows selecting the optimal parameter in function of a specific purpose. Using principle component analysis (PCA, twenty-three measured parameters were classified into three classes containing measures for size, shape and surface topology of the NM. Conclusion The presented method allows a detailed quantitative characterization of NM, like dispersions of precipitated and pyrogenic SAS based on the number-based distributions of their mean diameter, sphericity and shape factor.

  7. Time course of endotoxin-induced airways' inflammation in healthy subjects.

    Science.gov (United States)

    Doyen, Virginie; Kassengera, Zaina; Dinh, Duc Huy Phong; Michel, Olivier

    2012-02-01

    Few data are available on the kinetic of the airways' inflammation induced by inhaled endotoxin in a given subject. The purpose of this study was to evaluate in healthy subjects the time-related endotoxin-induced airways' inflammation. The cells counts from the induced-sputum were evaluated before, 6 and 24 h, and 7 days after an exposure to 20 mcg inhaled endotoxin, in eight pre-selected volunteers. To avoid interference of the induced-sputum procedure on the response to endotoxin, each time-point was evaluated in randomized order at 2-weeks interval after three separate inhalations of endotoxin. A significant rise of the relative number of lymphocytes (pinflammation after 6 h, peaked at 24 h and recovered after 7 days. When repeated endotoxin inhalations are used as a model of inflammation, a wash-out period of at least 7 days should be applied between each exposure in each subject.

  8. Comparative studies of endotoxin uptake by isolated rat Kupffer and peritoneal cells.

    OpenAIRE

    Fox, E S; Thomas, P; Broitman, S A

    1987-01-01

    The process of uptake of endotoxin by cells of the reticuloendothelial system is of current interest. Rabbit peritoneal macrophages have been used to study macrophage-endotoxin interactions and have suggested a receptor-mediated process. It is generally believed that the site of in vivo endotoxin clearance is the liver and that this clearance involves the Kupffer cell population. In the current report, the uptake characteristics of iodine-125-labeled Salmonella minnesota lipopolysaccharide (L...

  9. Treatment Characteristics of Polysaccharides and Endotoxin Using Oxygen Plasma Produced by RF Discharge

    Science.gov (United States)

    Kitazaki, Satoshi; Hayashi, Nobuya; Goto, Masaaki

    2010-10-01

    Treatment of polysaccharides and endotoxin were attempted using oxygen plasma produced by RF discharge. Oxygen radicals observed by optical light emission spectra are factors of decomposition of polysaccharides and endotoxin. Fourier transform infrared spectra indicate that most of chemical bonds in the polysaccharides are dissociated after irradiation of the oxygen plasma. Also, the decomposition rate of endotoxin was approximately 90% after irradiation of the oxygen plasma for 180 min.

  10. Peripherally administered orexin improves survival of mice with endotoxin shock

    Science.gov (United States)

    Ogawa, Yasuhiro; Irukayama-Tomobe, Yoko; Murakoshi, Nobuyuki; Kiyama, Maiko; Ishikawa, Yui; Hosokawa, Naoto; Tominaga, Hiromu; Uchida, Shuntaro; Kimura, Saki; Kanuka, Mika; Morita, Miho; Hamada, Michito; Takahashi, Satoru; Hayashi, Yu; Yanagisawa, Masashi

    2016-01-01

    Sepsis is a systemic inflammatory response to infection, accounting for the most common cause of death in intensive care units. Here, we report that peripheral administration of the hypothalamic neuropeptide orexin improves the survival of mice with lipopolysaccharide (LPS) induced endotoxin shock, a well-studied septic shock model. The effect is accompanied by a suppression of excessive cytokine production and an increase of catecholamines and corticosterone. We found that peripherally administered orexin penetrates the blood-brain barrier under endotoxin shock, and that central administration of orexin also suppresses the cytokine production and improves the survival, indicating orexin’s direct action in the central nervous system (CNS). Orexin helps restore body temperature and potentiates cardiovascular function in LPS-injected mice. Pleiotropic modulation of inflammatory response by orexin through the CNS may constitute a novel therapeutic approach for septic shock. DOI: http://dx.doi.org/10.7554/eLife.21055.001 PMID:28035899

  11. 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.

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

    Science.gov (United States)

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

    2011-01-01

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

  13. How does pyrogenic organic matter affect the N dynamic in agricultural soils? An incubation study

    Science.gov (United States)

    de La Rosa, José M.; Knicker, Heike

    2010-05-01

    Besides other environmental factors, N availability drives the carbon (C) and nitrogen (N) cycles in grasslands. Since grass-dominated ecosystems cover approximately 40% of the terrestrial surface and store more than 30% of global soil organic carbon (SOC), alterations to those ecosystems could have significant consequences and potential implications for global C and N cycles and climate (Schlesinger et al., 1990). Understanding the processes that govern the efficient cycling of nutrients through soil/plant systems remains an important topic to underpin the choice of strategies aimed at ensuring the long-term sustainability of ecosystems. In Mediterranean ecosystems, wild-fires occur frequently. Whereas factors such as water shortage or erosion contribute to reduced N-availability by lowering the litter input, burning additionally increase the refractory N and C-pools by charring litter and humic material (charred pyrogenic organic matter-PyOM) (Gonzalez-Pérez, 2004). In general, the addition of organic matter either as plant residues or farmyard manure has been shown to significantly increase biological activity, microbial biomass and enzyme activity in soil (Dick, 1992). Even in situations where microbial biomass appears to be unaffected, the activity of specific processes (e.g. N mineralization) can be significantly influenced by the addition of organic residues). However, little is known about the changes of the N cycle caused by the addition of PyOM. Therefore, the interest of our research was to study the impact of 15N enriched-biochars either alone or in conjunction with a 15N enriched fertilizer (K15NO3) on aggregate stability and organic carbon (C) and nitrogen (N) distribution among the different soil fractions. The latter may help to elucidate both, the quality of the stored organic matter and if the accumulation is related to interaction with the mineral matter. Therefore, biochar derived from grass material grown on 15N-enriched fertilizer was added

  14. Black (pyrogenic carbon in boreal forests: a synthesis of current knowledge and uncertainties

    Directory of Open Access Journals (Sweden)

    C. M. Preston

    2006-02-01

    Full Text Available The carbon (C cycle in boreal regions is strongly influenced by fire, which converts biomass and detrital C mainly to gaseous forms (CO2 and smaller proportions of CO and CH4, and some 1–7% of mass to pyrogenic C (PyC. PyC is mainly produced as solid charred residues, including visually-defined charcoal, and a black carbon (BC fraction chemically defined by its resistance to laboratory oxidation, plus much lower proportions of volatile soot and polycyclic aromatic hydrocarbons (PAHs. All PyC is characterized by fused aromatic rings, but varying in cluster sizes, and presence of other elements (N, O and functional groups. There are several reasons for current interest in defining more precisely the role of PyC in the C cycle of boreal regions. First, PyC is resistant to decomposition, and therefore contributes to very stable C pools in soils and sediments. Second, it influences soil processes, mainly through its sorption properties and cation exchange capacity, and third, soot aerosols absorb solar radiation and may contribute to global warming. However, there are large gaps in the basic information needed to address these topics. While charcoal is commonly defined by visual criteria, analytical methods for BC are mainly based on various measures of oxidation resistance, or on yield of benzenepolycarboxylic acids. These methods are still being developed, and capture different fractions of the PyC "continuum". There are few quantitative reports of PyC production and stocks in boreal forests (essentially none for boreal peatlands, and results are difficult to compare due to varying experimental goals and methods, as well as inconsistent terminology. There are almost no direct field measurements of BC aerosol production from boreal wildfires, and little direct information on rates and mechanisms for PyC loss. Structural characterization of charred biomass and forest floor from wildfires generally indicates a low level of

  15. Impact of pyrogenic organic matter decomposition and induced priming effect on soil C budget.

    Science.gov (United States)

    Maestrini, Bernardo; Abiven, Samuel

    2014-05-01

    Pyrogenic organic matter (PyOM) results from the incomplete combustion of biomass and may contribute to constitute an important fraction of soil C in forest and agricultural soils, in the form of charcoal (produced by wildfires) or biochar (anthropogenic). Although many evidences exist on the long mean residence time of PyOM there is still a large uncertainty on PyOM loss processes and rate and on possible induced priming effect on non-PyOM. Therefore determining PyOM mineralization rate, loss processes and possible induced priming effect on soil organic matter decomposition are key issues to understand the impact of PyOM on the carbon (C) cycle. We investigated the impact of PyOM on soil C budget by combining results from three independent studies: (i) a field study to investigate PyOM mineralization rate and the relative importance of PyOM loss processes, (ii) a PyOM and soil incubation experiment to correlate C and N mineralization rates, (iii) a review of the priming effect induced by PyOM on soil organic C. We employed 13C labelled pinewood-derived PyOM for the field experiment and 13C labelled ryegrass-derived PyOM in the incubation experiment to trace PyOM losses. In the field experiment it was observed that: (i) Pyrolysis process reduced pinewood decomposition by a factor of 60, (ii) leaching and translocation of fresh PyOM along the soil profile were negligible compared to losses as CO2. In the incubation experiment we found that ryegrass induced a two phase priming effect on native soil organic matter, with a positive priming effect followed by a negative priming effect phase, we also found that ryegrass-derived PyOM decomposition was much slower than pinewood one. The different decomposition rate results probably from the different aromaticity of the two PyOM together with the different set-up of the two experiments. Both the incubation experiment and the meta-analysis revealed that PyOM may induce a two-phase priming effect on native soil organic matter

  16. Black (pyrogenic) carbon: a synthesis of current knowledge and uncertainties with special consideration of boreal regions

    Science.gov (United States)

    Preston, C. M.; Schmidt, M. W. I.

    2006-09-01

    The carbon (C) cycle in boreal regions is strongly influenced by fire, which converts biomass and detrital C mainly to gaseous forms (CO2 and smaller proportions of CO and CH4), and some 1-3% of mass to pyrogenic C (PyC). PyC is mainly produced as solid charred residues, including visually-defined charcoal, and a black carbon (BC) fraction chemically defined by its resistance to laboratory oxidation, plus much lower proportions of volatile soot and polycyclic aromatic hydrocarbons (PAHs). All PyC is characterized by fused aromatic rings, but varying in cluster sizes, and presence of other elements (N, O) and functional groups. The range of PyC structures is often described as a continuum from partially charred plant materials, to charcoal, soot and ultimately graphite which is formed by the combination of heat and pressure. There are several reasons for current interest in defining more precisely the role of PyC in the C cycle of boreal regions. First, PyC is largely resistant to decomposition, and therefore contributes to very stable C pools in soils and sediments. Second, it influences soil processes, mainly through its sorption properties and cation exchange capacity, and third, soot aerosols absorb solar radiation and may contribute to global warming. However, there are large gaps in the basic information needed to address these topics. While charcoal is commonly defined by visual criteria, analytical methods for BC are mainly based on various measures of oxidation resistance, or on yield of benzenepolycarboxylic acids. These methods are still being developed, and capture different fractions of the PyC structural continuum. There are few quantitative reports of PyC production and stocks in boreal forests (essentially none for boreal peatlands), and results are difficult to compare due to varying experimental goals and methods, as well as inconsistent terminology. There are almost no direct field measurements of BC aerosol production from boreal wildfires, and

  17. Black (pyrogenic carbon: a synthesis of current knowledge and uncertainties with special consideration of boreal regions

    Directory of Open Access Journals (Sweden)

    C. M. Preston

    2006-01-01

    Full Text Available The carbon (C cycle in boreal regions is strongly influenced by fire, which converts biomass and detrital C mainly to gaseous forms (CO2 and smaller proportions of CO and CH4, and some 1–3% of mass to pyrogenic C (PyC. PyC is mainly produced as solid charred residues, including visually-defined charcoal, and a black carbon (BC fraction chemically defined by its resistance to laboratory oxidation, plus much lower proportions of volatile soot and polycyclic aromatic hydrocarbons (PAHs. All PyC is characterized by fused aromatic rings, but varying in cluster sizes, and presence of other elements (N, O and functional groups. The range of PyC structures is often described as a continuum from partially charred plant materials, to charcoal, soot and ultimately graphite which is formed by the combination of heat and pressure. There are several reasons for current interest in defining more precisely the role of PyC in the C cycle of boreal regions. First, PyC is largely resistant to decomposition, and therefore contributes to very stable C pools in soils and sediments. Second, it influences soil processes, mainly through its sorption properties and cation exchange capacity, and third, soot aerosols absorb solar radiation and may contribute to global warming. However, there are large gaps in the basic information needed to address these topics. While charcoal is commonly defined by visual criteria, analytical methods for BC are mainly based on various measures of oxidation resistance, or on yield of benzenepolycarboxylic acids. These methods are still being developed, and capture different fractions of the PyC structural continuum. There are few quantitative reports of PyC production and stocks in boreal forests (essentially none for boreal peatlands, and results are difficult to compare due to varying experimental goals and methods, as well as inconsistent terminology. There are almost no direct field measurements of BC aerosol production from boreal

  18. 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

  19. Host Immune Response to Bacterial Cyclic Diguanylic Acid (c-di-GMP)

    Science.gov (United States)

    2009-07-01

    Materials and Methods c-di-GMP and chemicals used The c-di-GMP (Fig. 1) used in these studies was synthesized and prepared as previously described (19–24...godeoxynucleotide (ODN) 2216 (25). LPS endotoxin (Limulus amebocyte lysate assay) test A dose of 500 M c-di-GMP was tested for the presence of LPS endotoxin using...inoculation, 100 CFU of S. aureus New- bould 305 (ATCC 29740) were injected into each gland and the coloniza- tion was allowed for 10 h. Raw bacterial CFU

  20. 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.

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

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Huizhi; Fan, Daidi, E-mail: fandaidi@nwu.edu.cn; Deng, Jianjun; Zhu, Chenghui; Hui, Junfeng; Ma, Xiaoxuan

    2014-09-01

    Protein preparation, which has active ingredients designated for the use of biomaterials and therapeutical protein, is obtained by genetic engineering, but products of genetic engineering are often contaminated by endotoxins. Because endotoxin is a ubiquitous and potent proinflammatory agent, endotoxin removal or depletion from protein is essential for researching any biomaterials. In this study, we have used Tris-acetate (TA) buffer of neutral pH value to evaluate endotoxins absorbed on the Pierce high-capacity endotoxin removal resin. The effects of TA buffer on pH, ionic strength, incubation time as well as human-like collagen (HLC) concentration on eliminating endotoxins are investigated. In the present experiments, we design an optimal method for TA buffer to remove endotoxin from recombinant collagen and use a chromogenic tachypleus amebocyte lysate (TAL) test kit to measure the endotoxin level of HLC. The present results show that, the endotoxins of HLC is dropped to 8.3 EU/ml at 25 mM TA buffer (pH 7.8) with 150 mM NaCl when setting incubation time at 6 h, and HLC recovery is about 96%. Under this experimental condition, it is proved to exhibit high efficiencies of both endotoxin removal and collagen recovery. The structure of treated HLC was explored by Transmission Electron Microscopy (TEM), demonstrating that the property and structure of HLC treated by TA buffer are maintained. Compared to the most widely used endotoxin removal method, Triton X-114 extraction, using TA buffer can obtain the non-toxic HLC without extra treatment for removing the toxic substances in Triton X-114. In addition, the present study aims at establishing a foundation for further work in laboratory animal science and providing a foundation for medical grade biomaterials. - Graphical abstract: The processes of endotoxins adsorbed from HLC. - Highlights: • TA buffer is a mild buffer system for endotoxins removal of HLC. • TA buffer may facilitate endotoxins adsorbed on the

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

    Science.gov (United States)

    Zhang, Huizhi; Fan, Daidi; Deng, Jianjun; Zhu, Chenghui; Hui, Junfeng; Ma, Xiaoxuan

    2014-09-01

    Protein preparation, which has active ingredients designated for the use of biomaterials and therapeutical protein, is obtained by genetic engineering, but products of genetic engineering are often contaminated by endotoxins. Because endotoxin is a ubiquitous and potent proinflammatory agent, endotoxin removal or depletion from protein is essential for researching any biomaterials. In this study, we have used Tris-acetate (TA) buffer of neutral pH value to evaluate endotoxins absorbed on the Pierce high-capacity endotoxin removal resin. The effects of TA buffer on pH, ionic strength, incubation time as well as human-like collagen (HLC) concentration on eliminating endotoxins are investigated. In the present experiments, we design an optimal method for TA buffer to remove endotoxin from recombinant collagen and use a chromogenic tachypleus amebocyte lysate (TAL) test kit to measure the endotoxin level of HLC. The present results show that, the endotoxins of HLC is dropped to 8.3EU/ml at 25 mM TA buffer (pH7.8) with 150 mM NaCl when setting incubation time at 6h, 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.

  3. Evolution of the surface polar character of pyrogenic silicas, with their grafting ratios by dimethylchlorosilane, studied by microcalorimetry.

    Science.gov (United States)

    Donnet, Jean-Baptiste; Ridaoui, Hassan; Balard, Henri; Barthel, Herbert; Gottschalk-Gaudig, Torsten

    2008-09-01

    The interactions of water, hexamethyldisiloxane, and dodecane with pyrogenic silica samples, modified by a controlled partial silylation with dimethyldichlorosilane, were studied by microcalorimetry and wettability measurements. The samples, having a coverage ratio lower than dimethylsilyl (DMS) monolayer capacity ( approximately 2.6 DMS/nm(2)), show a regular and linear decrease of their heat of immersion into water with the coverage ratio and correlate with the increase of residual silanol groups. Two critical coverage ratios were evidenced at about 25 and 50% of the DMS monolayer capacity, the grafted silica remaining hydrophilic, below 25% being strongly hydrophobic beyond. The heat of immersion into hexamethyldisiloxane decreases until 50% of the DMS monolayer whereas that of dodecane remains independent of the grafting ratio. This study demonstrates that the water/residual free silica surface plays the main role in the stabilization of the W/O Pickering's emulsions.

  4. 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

  5. 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.

  6. Enhancement of systemic and sputum granulocyte response to inhaled endotoxin in people with the GSTM1 null genotype

    Science.gov (United States)

    To determine if the GSTM1 null genotype is a risk factor for increased inflammatory response to inhaled endotoxin. Methods 35 volunteers who had undergone inhalation challenge with a 20 000 endotoxin unit dose of Clinical Center Reference Endotoxin (CCRE) were genotyped for the G...

  7. Concentrations and emissions of airborne endotoxins and microorganisms in livestock buildings in Northern Europe

    NARCIS (Netherlands)

    Seedorf, J.; Hartung, J.; Schröder, M.; Linkert, K.H.; Metz, J.H.M.; Groot Koerkamp, P.W.G.; Uenk, G.H.

    1998-01-01

    The concentration of airborne endotoxins and microorganisms in livestock buildings (cattle, pig, poultry) was surveyed in four European countries (England, The Netherlands, Denmark and Germany). Measurements were made during the day and night. The endotoxin concentrations were determined from dust s

  8. 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.

  9. 40 CFR 180.1107 - Delta endotoxin of Bacillus thuringiensis variety kurstaki encapsulated into killed Pseudomonas...

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Delta endotoxin of Bacillus... From Tolerances § 180.1107 Delta endotoxin of Bacillus thuringiensis variety kurstaki encapsulated into... Bacillus thuringiensis variety kurstaki encapsulated into killed Pseudomonas fluorescens is exempt from...

  10. Experimental design and Bayesian networks for enhancement of delta-endotoxin production by Bacillus thuringiensis.

    Science.gov (United States)

    Ennouri, Karim; Ayed, Rayda Ben; Hassen, Hanen Ben; Mazzarello, Maura; Ottaviani, Ennio

    2015-12-01

    Bacillus thuringiensis (Bt) is a Gram-positive bacterium. The entomopathogenic activity of Bt is related to the existence of the crystal consisting of protoxins, also called delta-endotoxins. In order to optimize and explain the production of delta-endotoxins of Bacillus thuringiensis kurstaki, we studied seven medium components: soybean meal, starch, KH₂PO₄, K₂HPO₄, FeSO₄, MnSO₄, and MgSO₄and their relationships with the concentration of delta-endotoxins using an experimental design (Plackett-Burman design) and Bayesian networks modelling. The effects of the ingredients of the culture medium on delta-endotoxins production were estimated. The developed model showed that different medium components are important for the Bacillus thuringiensis fermentation. The most important factors influenced the production of delta-endotoxins are FeSO₄, K2HPO₄, starch and soybean meal. Indeed, it was found that soybean meal, K₂HPO₄, KH₂PO₄and starch also showed positive effect on the delta-endotoxins production. However, FeSO4 and MnSO4 expressed opposite effect. The developed model, based on Bayesian techniques, can automatically learn emerging models in data to serve in the prediction of delta-endotoxins concentrations. The constructed model in the present study implies that experimental design (Plackett-Burman design) joined with Bayesian networks method could be used for identification of effect variables on delta-endotoxins variation.

  11. ENDOTOXIN RELEASE AND TUMOR-NECROSIS-FACTOR FORMATION DURING CARDIOPULMONARY BYPASS

    NARCIS (Netherlands)

    JANSEN, NJG; VANOEVEREN, W; GU, YJ; VANVLIET, MH; EIJSMAN, L; WILDEVUUR, CRH

    1992-01-01

    Endotoxin, when released into the systemic circulation during cardiopulmonary bypass (CPB), might induce activation of plasmatic systems and blood cells during CPB, in addition to a material-dependent blood activation during CPB. However, the role of endotoxin in the development of this so-called wh

  12. 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.

  13. Single session of Nd:YAG laser intracanal irradiation neutralizes endotoxin in dental root dentin

    Science.gov (United States)

    Archilla, José R. F.; Moreira, Maria S. N. A.; Miyagi, Sueli P. H.; Bombana, Antônio C.; Gutknecht, Norbert; Marques, Márcia M.

    2012-11-01

    Endotoxins released in the dental root by Gram-negative microorganisms can be neutralized by calcium hydroxide, when this medication is applied inside the root canal for at least seven days. However, several clinical situations demand faster root canal decontamination. Thus, for faster endotoxin neutralization, endodontists are seeking additional treatments. The in vitro study tested whether or not intracanal Nd:YAG laser irradiation would be able to neutralize endotoxin within the human dental root canal in a single session. Twenty-four human teeth with one root were mounted between two chambers. After conventional endodontic treatment, root canals were contaminated with Escherichia coli endotoxin. Then they were irradiated or not (controls) in contact mode with an Nd:YAG laser (1.5 W, 15 Hz, 100 mJ and pulse fluency of 124 J/cm2). The endotoxin activity was measured using the limulus lysate technique and data were statistically compared (p≤0.05). The concentration of active endotoxin measured in the negative control group was significantly lower than that of the positive control group (p=0.04). The concentrations of endotoxin in both irradiated groups were significantly lower than that of the positive control group (p=0.027) and similar to that of negative control group (p=0.20). A single session of intracanal Nd:YAG laser irradiation is able to neutralize endotoxin in the dental root tissues.

  14. Immunostimulatory Potential of β-Lactoglobulin Preparations: Effects Caused by Endotoxin Contamination

    DEFF Research Database (Denmark)

    Pedersen, Susanne Brix; Bovetto, L.; Fritsche, R.

    2003-01-01

    the immunomodulatory activity. Eventually, the immunostimulatory effect was found to be caused by endotoxin contamination.Conclusion: These results identify endotoxin as the main immunostimulatory component present in some commercial beta-lactoglobulin preparations. Moreover, the present study makes it evident...

  15. Surperoxide-mediated glomerulopathy in the endotoxin-treated pregnant rat

    NARCIS (Netherlands)

    Faas, MM; Schuiling, GA; Valkhof, N; Baller, JWF; Bakker, WW

    1998-01-01

    In the present study the role of superoxide in the glomerular damage in the low-dose endotoxin-infused pregnant rats was investigated. On day 14 of pregnancy, 12 rats were infused for 1 h with 1.0 mu g/kg bw endotoxin via a permanent jugular vein cannula. Of these rats, 6 were treated with SOD both

  16. p38-MAPK inhibition and endotoxin induced tubular dysfunction in men

    NARCIS (Netherlands)

    Zijlstra, JG; Tulleken, JE; Ligtenberg, JJM; de Boer, P; van der Werf, TS

    2004-01-01

    Background: To evaluate the possibility of preventing endotoxin induced renal damage by p38-MAPK inhibition in a human model. Design and Methods: Twenty-one healthy young male volunteers received 4 ng/kg Escherichia coli endotoxin as a single dose. Four groups of volunteers received an oral dose of

  17. 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

  18. An Endotoxin Tolerance Signature Predicts Sepsis and Organ Dysfunction at Initial Clinical Presentation

    Directory of Open Access Journals (Sweden)

    Olga M. Pena

    2014-11-01

    Interpretation: Our data support an updated model of sepsis pathogenesis in which endotoxin tolerance-mediated immune dysfunction (cellular reprogramming is present throughout the clinical course of disease and related to disease severity. Thus endotoxin tolerance might offer new insights guiding the development of new therapies and diagnostics for early sepsis.

  19. Insoluble glycogen, a metabolizable internal adsorbent, decreases the lethality of endotoxin shock in rats

    Directory of Open Access Journals (Sweden)

    S. Sipka

    1997-01-01

    Full Text Available Insoluble glycogen is an enzymatically modified form of naturally occurring soluble glycogen with a great adsorbing capacity. It can be metabolized by phagocytes to glucose. In this study we used insoluble glycogen intravenously in the experimental endotoxin shock of rats. Wistar male rats were sensitized to endotoxin by Pb acetate. The survival of rats were compared in groups of animals endotoxin shock treated and non-treated with insoluble glycogen. Furthermore, we have determined in vitro the binding capacity of insoluble glycogen for endotoxin, tumour necrosis factor alpha, interleukin-1 and secretable phospholipase A2. Use of 10 mg/kg dose of insoluble glycogen could completely prevent the lethality of shock induced by LD50 quantity of endotoxin in rats. All animals treated survived. Insoluble glycogen is a form of ‘metabolizable internal adsorbents’. It can potentially be used for treatment of septic shock.

  20. 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

    strains, and some of their pathogen-associated molecular patterns, were incubated apically on a confluent layer of intestinal epithelial cells (Caco-2), which were basolaterally co-cultured with human mononuclear leukocytes. Only Gram-negative bacteria having Enterobacteriaceae-type endotoxin (commensal...... fermenters (Lactobacillus spp.) did not stimulate leukocytes transepithelially. Endotoxin from E. coli and Salmonella enterica Typhimurium induced comparable transepithelial stimulation of leukocytes, but not endotoxin from B. vulgatus or lipoteichoic acid from E. faecalis. Endotoxin-binding agents...... (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...

  1. 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.

  2. Endotoxin Structures in the Psychrophiles Psychromonas marina and Psychrobacter cryohalolentis Contain Distinctive Acyl Features

    Directory of Open Access Journals (Sweden)

    Charles R. Sweet

    2014-07-01

    Full Text Available Lipid A is the essential component of endotoxin (Gram-negative lipopolysaccharide, a potent immunostimulatory compound. As the outer surface of the outer membrane, the details of lipid A structure are crucial not only to bacterial pathogenesis but also to membrane integrity. This work characterizes the structure of lipid A in two psychrophiles, Psychromonas marina and Psychrobacter cryohalolentis, and also two mesophiles to which they are related using MALDI-TOF MS and fatty acid methyl ester (FAME GC-MS. P. marina lipid A is strikingly similar to that of Escherichia coli in organization and total acyl size, but incorporates an unusual doubly unsaturated tetradecadienoyl acyl residue. P. cryohalolentis also shows structural organization similar to a closely related mesophile, Acinetobacter baumannii, however it has generally shorter acyl constituents and shows many acyl variants differing by single methylene (-CH2- units, a characteristic it shares with the one previously reported psychrotolerant lipid A structure. This work is the first detailed structural characterization of lipid A from an obligate psychrophile and the second from a psychrotolerant species. It reveals distinctive structural features of psychrophilic lipid A in comparison to that of related mesophiles which suggest constitutive adaptations to maintain outer membrane fluidity in cold environments.

  3. 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.

  4. Evaluation and Safety Control Mechanism of Endotoxin Detection in Whole Blood and Blood Components%全血和成分血内毒素检测评估及其安全控制机制的评价

    Institute of Scientific and Technical Information of China (English)

    鲍自谦; 刘怡

    2013-01-01

    Objective Through dynamic monitoring bacterial endotoxin and cultivating whole blood and blood components,consistence between endotoxin level and bacterial culture results,association between the clinic fever reaction and blood endotoxin were evaluated.Furthermore,the effectiveness of safety control mechanism on bacterium and endotoxin was assessed.Methods From March to April 2009,a total of 300 blood samples of volunteer donors from our blood center were included in this study.Based on the sample varieties,all of these samples were divided into 3 groups:the whole blood group (n=50),suspended red blood cell (RBC) components group (n=150) and platelet (PLT) components group (n=100).Meanwhile 361 healthy donors from the street during January to February 2009,were chosen as a control group,and their endotoxin results would be used as normal reference range.Dynamic turbidity was used as the quantitative detection method of bacterial endotoxin.Endotoxin content in each group was individually detected during their blood preservations.And the endotoxin content results of each group in different blood preservation period and their bacterial cultivating results were analyzed by variance of repeated measurement data.Moreover,the samples from clinical transfusion fever reaction were detected for endotoxin level.Results Normal blood endotoxin reference range from the control group was (0-0.193) EU/mL.PLT suspension group (W=0.635,P =0.0585),and suspended RBC group (W =0.334,P =0.583) had no significant differences in endotoxin concentration of three periods.There were significant differences in endotoxin in whole blood group of three periods (W=0.860,P=0.014).The endotoxin level of whole blood and blood components was (0.005-0.188) EU/mL,and all bacterial cultivating results were negative.Endotoxin concentration in blood samples of transfusion fever reaction was (0.023-0.145) EU/mL.Conclusions While cultivating results of whole blood and blood components were negative

  5. 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.

  6. Detection of endotoxin using a photonic crystal nanolaser

    Science.gov (United States)

    Takahashi, Daichi; Hachuda, Shoji; Watanabe, Takumi; Nishijima, Yoshiaki; Baba, Toshihiko

    2015-03-01

    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. The ability of endotoxin adsorption during a longer duration of direct hemoperfusion with a polymyxin B-immobilized fiber column in patients with septic shock.

    Science.gov (United States)

    Shimizu, Tomoharu; Obata, Toru; Sonoda, Hiromichi; Akabori, Hiroya; Tabata, Takahisa; Eguchi, Yutaka; Endo, Yoshihiro; Tani, Tohru

    2013-12-01

    The patients' hemodynamic conditions of septic shock due to intra-abdominal infection were improved by the longer duration of direct hemoperfusion with a polymyxin B-immobilized fiber column (PMX), reducing plasma endotoxins measured by the novel endotoxin detection method, named endotoxin scattering photometry (ESP) method; however, turbidimetric method could not detect endotoxins. We also observed the reduction in the endotoxin after passing through column by ESP method even after the longer duration of PMX. ESP method may more sensitively detect endotoxins than the ordinary turbidimetric method. Moreover, we demonstrated the ability of endotoxin adsorption in spite of the longer duration of PMX.

  8. 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

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

    Directory of Open Access Journals (Sweden)

    S K Makkar

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

  10. 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

  11. Bacterial Vaginosis

    Science.gov (United States)

    ... Issues > Conditions > Sexually Transmitted > Bacterial Vaginosis Health Issues Listen Español Text Size Email Print Share Bacterial Vaginosis Page Content Bacterial vaginosis (BV) is the most common vaginal infection in sexually active teenaged girls . It appears to be caused by ...

  12. Residential indoor and outdoor coarse particles and associated endotoxin exposures

    Science.gov (United States)

    Wheeler, Amanda J.; Dobbin, Nina A.; Lyrette, Ninon; Wallace, Lance; Foto, Mark; Mallick, Ranjeeta; Kearney, Jill; Van Ryswyk, Keith; Gilbert, Nicolas L.; Harrison, Ian; Rispler, Kathleen; Héroux, Marie-Eve

    2011-12-01

    There is a growing body of evidence demonstrating that coarse particles (PM 10-2.5) have detrimental impacts upon health, especially for respiratory effects. There are limited data available for indoor residential exposures. Some data exist regarding the composition of this PM size fraction with emphasis on crustal elements and biological components. This study includes data from 146 homes sampled in Regina, Saskatchewan (SK) where 5-day integrated concurrent monitoring of indoor and outdoor coarse particles was conducted during the winter and summer of 2007. The coarse particle filters were subsequently analysed for endotoxin content to determine the contribution of this compound. Winter indoor geometric mean concentrations of coarse particles exceeded outdoor concentrations (3.73 μg m -3 vs 2.49 μg m -3; paired t-test p Endotoxin concentrations (EU μg -1) were lower indoors than outdoors in both seasons. Spatial variability of ambient coarse particles was assessed to determine the suitability of using a single monitoring station within a city to estimate exposure. The coefficients of variation between homes sampled simultaneously and the central monitoring station were calculated (median COV in summer = 15% and winter = 24%) and showed significant variability by week, especially during the summer months, suggesting a single site may be insufficient for characterizing exposure. Future studies should consider daily measurements per home to understand shorter term exposures and day to day variability of these pollutants.

  13. Combining charcoal and elemental black carbon analysis in sedimentary archives : implications for past fire regimes, the pyrogenic carbon cycle, and the human-climate interactions

    OpenAIRE

    2010-01-01

    This paper addresses the quantification of combustion-derived products in oceanic and continental sediments by optical and chemical approaches, and the interest of combining such methods for reconstructing past biomass burning activity and the pyrogenic carbon cycle. In such context, the dark particles >0.2 mu m(2) remaining after the partial digestion of organic matter are optically counted by automated image analysis and defined as charcoal, while the elemental carbon remaining after therma...

  14. [Carriage of Streptococcus pyogenes in primary school children: M-protein types, pyrogenic toxin genes, and investigation of the clonal relationships between the isolates].

    Science.gov (United States)

    Otlu, Barış; Karakurt, Cemşit; Bayındır, Yaşar; Kayabaş, Üner; Yakupoğulları, Yusuf; Gözükara Bağ, Harika

    2015-07-01

    M-protein and pyrogenic toxins are the most important virulence factors of Streptococcus pyogenes, and they play significant role in the pathophysiology of acute rheumatoid fever and scarlet fever, respectively. In this study, the pharyngeal carriage of S.pyogenes of the primary school children, clonal relationship of the strains, M-protein types, and the presence of pyrogenic toxin genes were aimed to be investigated. A total of 668 throat cultures obtained from children (age range: 6-16 years) in two primary schools in our region, were included in the study. The clonal relationships of the isolated group A streptococci (GAS) strains were investigated by DiversiLab assay (BioMérieux, France), and the clonal relatedness was confirmed by pulsed-field gel electrophoresis (PFGE) method. M-protein (emm) typing was performed by DNA sequencing as suggested by Centers for Disease Control and Prevention (CDC). The genes encoding pyrogenic toxins, speA and speC, were investigated by an in-house multiplex polymerase chain reaction (PCR) method. S.pyogenes was isolated from 134 (20.05%) of the throat samples. The GAS carriage rate of the students aged ≥10 was statistically higher than those 7-9 years age group (%22 vs %16.4, pvaccine was determined to be over 90% with respect to M-protein types. Since the pyrogenic toxin-encoding genes were found in one fifth of the isolates from the studied subjects, we concluded that the carrier population may also have high risk for scarlet fever. We also concluded that, the clonal relationship ratio determined among the isolates may be a risk in school transmission of GAS.

  15. 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.

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

    Science.gov (United States)

    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, 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. PMID:16722306

  17. 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.

  18. 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. 

  19. Effect of dialyser membranes on extracellular and intracellular granulocyte and monocyte activation in ex vivo pyrogen-free conditions.

    Science.gov (United States)

    Mahiout, A; Courtney, J M

    1994-10-01

    This study examined effects of blood-contacting materials on the monocyte reaction following the first contact of human blood with hollow fibre dialyser membranes under pyrogen-free conditions. Membrane materials were the unchanged regenerated cellulose, the synthetic polysulphone (PS), a positively charged diethylaminoethyl cellulose (DEAE-C), the negatively charged carboxymethyl cellulose (CMC) and acrylonitrile copolymer (AN). The experimental system involved perfusion with human fresh venous blood through different modules containing the materials in the form of hollow fibre membranes. Extracellular and intracellular aspects of blood reactions after the first contact with the materials were investigated in Ficoll-separated granulocytes and peripheral blood mononuclear cells. Investigations were done by release reactions of platelet activating factor (PAF), oxygen radical (O2-), leukotriene B4, prostaglandin E2 (PGE2) and cytokines (IL-1 beta, TNF-alpha, IL-6). The intracellular activation of peripheral blood mononuclear cells was done by mRNA transcription of IL-1 beta, TNF-alpha, IL-6, IL-8 and beta 2-microglobulin (beta 2-MG). From the set of parameters, release reactions were only measurable for PAF, PGE2 and O2- if a second stimulus (phorbol myristate acetate, lipopolysaccharide, zymosan and calcium ionophore) was used after blood-membrane interaction. Although the extent of the release reaction was weak, negatively charged membranes were, in general, more active. All dialysers exhibited the same increase in beta 2-MG mRNA transcription, suggesting that all blood-contacting membranes initiate the gene expression of beta 2-MG at the same level. TNF-alpha, IL-6, IL-1 beta and IL-8 mRNAs were demonstrated in the AN and CMC membranes rather than the other materials, which exhibit a lower transcription than the tubing set. As has been found, an enhanced generation of PGE2 for both CMC and AN membranes supports, therefore, the concept of an effect of the negative

  20. 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.

  1. 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.

  2. 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.

  3. The molecular adsorption-type endotoxin detection system using immobilized ɛ-polylysine

    Science.gov (United States)

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

    2007-12-01

    Hemodialysis for chronic renal failure is the most popular treatment method with artificial organs. However, hemodialysis patients must continue the treatment throughout their life, the results of long term extracorporeal dialysis, those patients develop the various complications and diseases, for example, dialysis amyloidosis etc. Dialysis amyloidosis is one of the refractory complications, and endotoxin is thought to be the most likely cause of it, recently. Endotoxin is one of the major cell wall components of gram-negative bacteria, and it has various biological activities. In addition, it is known that a mount of endotoxin exists in living environment, and medicine is often contaminated with endotoxin. When contaminated dialyzing fluids are used to hemodialysis, above-mentioned dialysis amyloidosis is developed. Therefore, it is important that the detection and removal of endotoxin from dialyzing fluids. Until now, the measurement methods using Limulus Amebosyte Lysate (LAL) reagent were carried out as the tests for the presence of endtoxin. However, these methods include several different varieties of measurement techniques. The following are examples of them, gelatinization method, turbidimetric assay method, colorimetric assay method and fluoroscopic method. However, these techniques needed 30-60 minutes for the measurement. From these facts, they are not able to use as a "real-time endotoxin detector". The detection of endotoxin has needed to carry out immediately, for that reason, a new detection method is desired. In this research, we focused attention to adsorption reaction between ɛ-polylysine and endotoxin. ɛ-polylysine has the structure of straight chain molecule composed by 25-30 residues made by lysine, and it is used as an antimicrobial agent, moreover, cellulose beads with immobilized ɛ-polylysine is used as the barrier filter for endotoxin removal. The endotoxin is adsorbed to immobilized ɛ-polylysine, as the result of this reaction, the

  4. Lack of spatial variation of endotoxin in ambient particulate matter across a German metropolitan area

    Science.gov (United States)

    Morgenstern, Verena; Carty, Cara L.; Gehring, Ulrike; Cyrys, Josef; Bischof, Wolfgang; Heinrich, Joachim

    In this study, we describe the spatial variation of endotoxin across an urban setting using Geographic Information Systems (GIS) methods. We also identify potential sources of endotoxin that account for between-site variability and compare endotoxin levels in particulate matter with a 50% aerodynamic cut-off diameter of 2.5 μm (PM 2.5) and of 10 μm (PM 10). In 1999-2000, we collected PM 2.5 and PM 10 in Munich urban air and measured soluble endotoxin concentrations in both particle fractions. Using Teflon filters and Harvard impactors, PM 2.5 was collected at 40 outdoor monitoring sites across Munich and PM 10 at a subset of these sites ( n=12). Approximately four samples were collected at each site for a total of 158 PM 2.5 samples and 48 PM 10 samples. We visited and characterized the surrounding 100 m of each site for potential endotoxin sources. The geometric mean endotoxin concentration for all sites was 1.46 EU mg -1 PM 2.5 (95% confidence intervals (CI): 1.21-1.77) and at the subset of the sites was 1.30 EU mg -1 PM 2.5 (95% CI: 1.01-1.67 EU mg -1 PM 2.5). Endotoxin levels in PM 10 were higher, 3.91 EU mg -1 PM 10 (95% CI: 3.03-5.03 EU mg -1 PM 10), than in PM 2.5 and were moderately correlated, r=0.51. All endotoxin concentrations measured in this study were endotoxin levels (MR: 1.30 for EU mg -1 PM 2.5 and 1.13 for EU m -3 PM 2.5) than sites with no identified sources. Based on the ranges of endotoxin levels at the different sites, we found very little spatial variation in ambient endotoxin concentrations across the metropolitan area of Munich using inverse distance weighting method (IDW) methods ( R2=0.013 for EU mg -1 PM 2.5 and R2=0.020 for EU m -3 PM 2.5). Potential sources of endotoxin surrounding the sites only partly explained the variation seen.

  5. Labile pyrogenic dissolved organic carbon in major Siberian Arctic rivers: Implications for wildfire-stream metabolic linkages

    Science.gov (United States)

    Myers-Pigg, Allison N.; Louchouarn, Patrick; Amon, Rainer M. W.; Prokushkin, Anatoly; Pierce, Kayce; Rubtsov, Alexey

    2015-01-01

    burning produces a spectrum of thermally altered materials that releases pyrogenic carbon (PyC) to terrestrial, atmospheric, and aquatic systems. Most studies focus on the refractory end of the PyC spectrum, derived from middle- to high-temperature combustion. Low-temperature PyC is produced during wildfires and has been found to be particularly labile and water soluble. Here we find that in each of the major Siberian watersheds, low-temperature fire-derived biomarkers are present in detectable concentrations during all flow regimes of the 2004-2006 sampling period, confirming that PyC is an intrinsic component of the dissolved organic carbon (DOC) pool mobilized by hydrologic events. Gymnosperm combustion, from the southern portions of these watersheds, is the primary source of this Py-DOC input. Using first-order degradation rates and transit times of water through these rivers, about half of the total estimated flux of this material may be remineralized during transport from fire source to river mouth (20-40 days), demonstrating the input of a labile source of PyC to these watersheds.

  6. Source Material and Concentration of Wildfire-Produced Pyrogenic Carbon Influence Post-Fire Soil Nutrient Dynamics

    Directory of Open Access Journals (Sweden)

    Lucas A. Michelotti

    2015-04-01

    Full Text Available Pyrogenic carbon (PyC is produced by the thermal decomposition of organic matter in the absence of oxygen (O. PyC affects nutrient availability, may enhance post-fire nitrogen (N mineralization rates, and can be a significant carbon (C pool in fire-prone ecosystems. Our objectives were to characterize PyC produced by wildfires and examine the influence that contrasting types of PyC have on C and N mineralization rates. We determined C, N, O, and hydrogen (H concentrations and atomic ratios of charred bark (BK, charred pine cones (PC, and charred woody debris (WD using elemental analysis. We also incubated soil amended with BK, PC, and WD at two concentrations for 60 days to measure C and N mineralization rates. PC had greater H/C and O/C ratios than BK and WD, suggesting that PC may have a lesser aromatic component than BK and WD. C and N mineralization rates decreased with increasing PyC concentrations, and control samples produced more CO2 than soils amended with PyC. Soils with PC produced greater CO2 and had lower N mineralization rates than soils with BK or WD. These results demonstrate that PyC type and concentration have potential to impact nutrient dynamics and C flux to the atmosphere in post-fire forest soils.

  7. Collaborative study for the establishment of the WHO 3(rd) International Standard for Endotoxin, the Ph. Eur. endotoxin biological reference preparation batch 5 and the USP Reference Standard for Endotoxin Lot H0K354.

    Science.gov (United States)

    Findlay, L; Desai, T; Heath, A; Poole, S; Crivellone, M; Hauck, W; Ambrose, M; Morris, T; Daas, A; Rautmann, G; Buchheit, K H; Spieser, J M; Terao, E

    2015-01-01

    An international collaborative study was organised jointly by the World Health Organization (WHO)/National Institute for Biological Standards and Control (NIBSC), the United States Pharmacopeia (USP) and the European Directorate for the Quality of Medicines & HealthCare (EDQM/Council of Europe) for the establishment of harmonised replacement endotoxin standards for these 3 organisations. Thirty-five laboratories worldwide, including Official Medicines Control Laboratories (OMCLs) and manufacturers enrolled in the study. Three candidate preparations (10/178, 10/190 and 10/196) were produced with the same material and same formulation as the current reference standards with the objective of generating a new (3(rd)) International Standard (IS) with the same potency (10 000 IU/vial) as the current (2(nd)) IS, as well as new European Pharmacopoeia (Ph. Eur.). and USP standards. The suitability of the candidate preparations to act as the reference standard in assays for endotoxin performed according to compendial methods was evaluated. Their potency was calibrated against the WHO 2(nd) IS for Endotoxin (94/580). Gelation and photometric methods produced similar results for each of the candidate preparations. The overall potency estimates for the 3 batches were comparable. Given the intrinsic assay precision, the observed differences between the batches may be considered unimportant for the intended use of these materials. Overall, these results were in line with those generated for the establishment of the current preparations of reference standards. Accelerated degradation testing of vials stored at elevated temperatures supported the long-term stability of the 3 candidate preparations. It was agreed between the 3 organisations that batch 10/178 be shared between WHO and EDQM and that batches 10/190 and 10/196 be allocated to USP, with a common assigned value of 10 000 IU/vial. This value maintains the continuity of the global harmonisation of reference materials and

  8. Peripheral bacterial endotoxin administration triggers both memory consolidation and reconsolidation deficits in mice.

    Science.gov (United States)

    Kranjac, Dinko; McLinden, Kristina A; Deodati, Lauren E; Papini, Mauricio R; Chumley, Michael J; Boehm, Gary W

    2012-01-01

    Peripherally administered inflammatory stimuli, such as lipopolysaccharide (LPS), induce the synthesis and release of proinflammatory cytokines and chemokines in the periphery and the central nervous system, and trigger a variety of neurobiological responses. Indeed, prior reports indicate that peripheral LPS administration in rats disrupts contextual fear memory consolidation processes, potentially due to elevated cytokine expression. We used a similar, but partially olfaction-based, contextual fear conditioning paradigm to examine the effects of LPS on memory consolidation and reconsolidation in mice. Additionally, interleukin-1β (IL-1β), brain-derived neurotrophic factor (BDNF), and zinc finger (Zif)-268 mRNA expression in the hippocampus and the cortex, along with peripheral cytokines and chemokines, were assessed. As hypothesized, LPS administered immediately or 2 h, but not 12 h, post-training impaired memory consolidation processes that support the storage of the conditioned contextual fear memory. Additionally, as hypothesized, LPS administered immediately following the fear memory trace reactivation session impaired memory reconsolidation processes. Four hours post-injection, both central cytokine and peripheral cytokine and chemokine levels were heightened in LPS-treated animals, with a simultaneous decrease in BDNF, but not Zif-268, mRNA. Collectively, these data reinforce prior work showing LPS- and cytokine-related effects on memory consolidation, and extend this work to memory reconsolidation.

  9. Dose-Dependent Changes in the Antigenicity of Bacterial Endotoxin Exposed to Ionizing Radiation

    Science.gov (United States)

    1987-01-01

    M9lt6 LOS %%ere raised by immtunizing with purilied LOS for the elect rophoret ie blotting of LI’S antigens. LPS were transferred as described before 0 1...Traits- Blot cell. After treating the nitrocellulose membrane with 3%ý Hiomedical. Division of Marine Colloids. Inc.. Rockland. MEF) made tip gelatin in...uthrs han Mis T res Wison or ieroriing107. 137. inimunocfectrophoresis. 19. Land). M.. Johnson, A. G.. Wester , MI. E. and Sagin, J. F. (1955). This work

  10. Analysis of the Cellular and Molecular Mechanisms Which Underlie Sensitivity to Bacterial Endotoxin and Early Tolerance

    Science.gov (United States)

    1992-06-24

    1990). It is the O-antigen that confers antigenic and serological specificity on the bacterium (Luderitz et a1., 1982; 1987; reviewed in Brade et a1...Munford, 1983; Munford and Hall, 1985; 1986). AOAH enzyme assays were kindly performed by David Ginkel, in the laboratory of Robert S. Munford...MPL for 1 hour at 370 C in 6% C02. Affinity-purified rabbit anti-5D3 antibody (RaSD3) was provided by David 41 Morrison (University of Kansas

  11. Detecting endotoxin activity in bovine serum using an automated testing system.

    Science.gov (United States)

    Suzuki, Kazuyuki; Shimamori, Toshio; Sato, Ayano; Tsukano, Kenji; Tsuchiya, Masakazu; Lakritz, Jeffrey

    2015-08-01

    The aim of the present study was to compare the ability of the commercially available portable test system (PTS(TM)) to detect endotoxin activity in bovine serum, with that of the traditional LAL-kinetic turbidimetric (KT) and chromogenic (KC) assays. Prior to testing, serum samples, which were obtained from endotoxin-challenged cattle, were diluted 1:20 in endotoxin-free water and heated to 80°C for 10 min. The performance of the PTS(TM) was not significantly different from that of the traditional LAL-based assays. The results using PTS(TM) correlated with those using KT (r(2)=0.963, PPTS(TM) could be applied as a simplified system to assess endotoxin activity in bovine serum.

  12. 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.

  13. Evaluation Of Airborne Endotoxin Concentrations Associated With Management Of Crop Grown On Applied Biosolids

    Science.gov (United States)

    Public health concerns have been expressed regarding inhalation exposure associated with the application of biosolids on cropland, which is due to the potential aerosolization of microorganisms, cell wall products, volatile chemicals, and nuisance odors. Endotoxin is a component...

  14. Effect of plasmapheresis on the immune system in endotoxin-induced sepsis

    DEFF Research Database (Denmark)

    Toft, P; Schmidt, R; Broechner, A C

    2008-01-01

    BACKGROUND: It has been proposed that plasmapheresis is most effective when applied early in Gram-negative sepsis. We therefore studied the effect of early plasmapheresis on immunity in experimental Escherichia coli endotoxin-induced sepsis. METHODS: 20 pigs received 30 microg/kg of E. coli...... endotoxin. 40 min later, half of the pigs were treated with plasmapheresis which lasted 4 h. The adhesion molecules, the oxidative burst, the number of neutrophils in blood and lungs, and cytokines were measured. RESULTS: Infusion of endotoxin was associated with activation of adhesion molecules increased...... oxidative burst, increased concentration of cytokine, and accumulation of granulocytes in lung tissue. Plasmapheresis reduced the oxidative burst, and there was a tendency towards a reduced accumulation of granulocytes in the lung. CONCLUSION: Though plasmapheresis was initiated early after the endotoxin...

  15. Sensitisation to common allergens and respiratory symptoms in endotoxin exposed workers: a pooled analysis

    DEFF Research Database (Denmark)

    Basinas, Ioannis; Schlünssen, Vivi; Heederik, Dick

    2012-01-01

    Objective To test the hypotheses that current endotoxin exposure is inversely associated with allergic sensitisation and positively associated with non-allergic respiratory diseases in four occupationally exposed populations using a standardised analytical approach. Methods Data were pooled from...... 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...... Current endotoxin exposure was dose-dependently associated with a reduced prevalence of allergic sensitisation (ORs of 0.92, 0.81 and 0.66 for low mediate, high mediate and high exposure) and hay fever (ORs of 1.16, 0.81 and 0.58). Endotoxin exposure was a risk factor for organic dust toxic syndrome...

  16. Routine limulus amoebocyte lysate (LAL) test for endotoxin determination in milk using a Toxinometer ET-201.

    Science.gov (United States)

    Mottar, J; De Block, J; Merchiers, M; Vantomme, K; Moermans, R

    1993-05-01

    A rapid method of performing the Limulus amoebocyte lysate (LAL) test in milk is proposed using the Toxinometer ET-201. This instrument measured the increase in turbidity due to the interaction between the endotoxins of the Gram-negative bacteria and the LAL reagent, monitored the ratio Rt of the sequential to the initial transmission at 12 s intervals and quantified endotoxins by determination of the reaction time Tr required to obtain a 5% decrease in Rt. There was a good correlation between the toxinometrically determined endotoxin concentrations and the number of Gram-negative bacteria (SD, 0.18 log(plate count units)), and the repeatability (CV, 6-10%) was high. The assay may be useful for screening raw materials for UHT milk production, as the endotoxin content of the raw material is related to the rest proteinase activity in the UHT milk.

  17. 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.

  18. 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

    During "nondamaging" exercise, skeletal muscle markedly releases interleukin (IL)-6, and it has been suggested that one biological role of this phenomenon is to inhibit the production of tumor necrosis factor (TNF)- alpha, which is known to cause pathogenesis such as insulin resistance...... 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...... lipopolysaccharide endotoxin (0.06 ng/kg) i.v. to induce low-grade inflammation. In CON, plasma TNF-alpha increased significantly in response to endotoxin. In contrast, during EX, which resulted in elevated IL-6, and rhIL-6, the endotoxin-induced increase in TNF-alpha was totally attenuated. In conclusion, physical...

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

    DEFF Research Database (Denmark)

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

    2008-01-01

    Background Endotoxins are common contaminants in allergen preparations and affect antigen-specific cellular responses. Distinct effects of endotoxin on cells in human umbilical cord and adult blood are poorly defined. Objectives To examine the effect of endotoxins in allergen preparations...... on cellular responses in human cord and peripheral blood (PB). Methods The endotoxin content in beta lactoglobulin (BLG), the peanut allergen Ara h 1 and the major birch pollen allergen Bet v 1 was assessed. Proliferation and cytokine response of mononuclear cells towards contaminated and lipopolysaccharide...... (LPS)-free allergens were evaluated at different time-points. Fractions of contaminated BLG were generated and assayed on their immuno-stimulatory capacity. The involvement of toll-like receptor (TLR) 2 and 4 was investigated by blocking antibodies and TLR-transfected human embryonic kidney cells...

  20. Efficient endotoxin removal with a new sanitizable affinity column: Affi-Prep Polymyxin.

    Science.gov (United States)

    Talmadge, K W; Siebert, C J

    1989-08-04

    A new affinity column packing for removal of endotoxins has been prepared by coupling USP drug-quality polymyxin B to Affi-Prep, a novel synthetic macroporous polymer. Affi-Prep Polymyxin binds endotoxins from a number of different strains of gram-negative bacteria. Endotoxin binding is not significantly affected by 10 mg/ml of bovine serum albumin or human immunoglobulin G, by 1 mg/ml sodium dodecyl sulphate, or by 1 mg/ml deoxycholate. Affi-Prep Polymyxin is stable to treatment with 1.0 M sodium hydroxide, an important property for sanitizing the resin. The resin shows a high ligand stability, since no leakage of polymyxin B from the packing could be detected. Affi-Prep Polymyxin exhibited the highest endotoxin binding efficiency when compared with several commercial agarose affinity packings.

  1. Effect of Solcoseryl on the clinical course of experimental Escherichia coli-endotoxin mastitis.

    Science.gov (United States)

    Ziv, G; Jöchle, W

    1981-09-01

    Experimental Escherichia coli-endotoxin mastitis was induced in a single quarter in twenty-eight cows. The inflamed quarters of six of the cows were treated with 50 mg Solcoseryl infusion in an oil base starting 4-6 h after endotoxin infusion, and four additional infusions were given at 12-h intervals. The inflamed quarters of eight of the cows were similarly treated with 200 mg Solcoseryl whereas the remaining fourteen cows served as control. Solcoseryl treatment did not alter the systemic or local course of acute mastitis. Local reaction, as assessed by the California Mastitis Test (CMT), however, subsided considerably sooner in quarters treated with Solcoseryl, and the effect of Solcoseryl in hastening udder tissue repair processes appeared to be dose dependent. The 200 mg dose of Solcoseryl resulted in pre-endotoxin CMT scores as early as 6 days after endotoxin infusion.

  2. 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.

  3. 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%.

  4. Endotoxin Is Not Essential for the Development of Cockroach Induced Allergic Airway Inflammation

    OpenAIRE

    2012-01-01

    Purpose Cockroach (CR) is an important inhalant allergen and can induce allergic asthma. However, the mechanism by which CR induces airway allergic inflammation and the role of endotoxin in CR extract are not clearly understood in regards to the development of airway inflammation. In this study, we evaluated whether endotoxin is essential to the development of CR induced airway allergic inflammation in mice. Materials and Methods Airway allergic inflammation was induced by intranasal administ...

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

    OpenAIRE

    2011-01-01

    Abstract 1. Endotoxins as components of organic dust may have adverse effects on the respiratory health of workers in poultry buildings. The move towards more welfare-friendly housing systems for layers may increase worker exposure to air contaminants due to the use of litter. 2. The endotoxin concentrations in the inhalable fraction of airborne dust (below 100 ?m) from cage and alternative system henhouses (on-floor, free range and aviaries) were compared under both experiment...

  6. Anti-ocular-inflammatory Effects of Salvia hypoleuca Extract on Rat Endotoxin-Induced Uveitis

    OpenAIRE

    Jasem Estakhr; Nasim Javdan

    2012-01-01

    Salvia hypoleuca is used in Iranian traditional medicine as an agent for treatment of some diseases and troubles. In this study, attention was focused on the antioxidant effect of Salvia hypoleuca. The aim of the present study was to determine the effects of Salvia hypoleuca extract on Endotoxin-Induced Uveitis (EIU) in rats. In addition, the endotoxin-induced expression of the inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 proteins was determined in a mouse macrophage cell...

  7. Accelerated ovum transport in rabbits induced by endotoxin II. Changes in oviductal smooth muscle activity.

    Science.gov (United States)

    Hodgson, B J; Harper, M J; Valenzuela, G

    1978-01-01

    Oviductal mortility, measured with open-ended perfused catheters in anesthetized animals injected with human Chorionic Gonadotropin (hCG), is depressed 2 h following endotoxin injection and returns to control levels by 3 h after endotoxin injection. This decrease in motility is prevented by indomethacin. Endotoxin did not affect spontaneous or phenylephrine (PE)-induced contractions of oviduct when it was added to the bathing medium of in vitro tissues. Oviductal segments removed 2 h after endotoxin (26 h after hCG) showed electrical activity confined to the ampullary-isthmic-junction (AIJ), where ova were located; the dose-response curve for PE was shifted to the right and the maximum contraction was depressed. Activity of tissues removed 4 h after endotoxin more closely resembled control tissues except that the maximum contraction to PE was depressed, ova had passed out of the oviduct and a proovarian bias in the isthmus was not present. The response of the oviduct to prostaglandins (PGs) in vivo is critically dependent on the previous exposure to PGs. In endotoxin-treated animals PGE then PGF levels increase and the decrease in motility coincides with increased PGE levels, but accelerated ovum transport with the return of motility and activation of the isthmus.

  8. Endotoxin-induced maturation of monocytes in preterm fetal sheep lung.

    Science.gov (United States)

    Kramer, Boris W; Joshi, Shubhada N; Moss, Timothy J M; Newnham, John P; Sindelar, Richard; Jobe, Alan H; Kallapur, Suhas G

    2007-08-01

    The fetal lung normally contains immature monocytes and very few mature macrophages. The chorioamnionitis frequently associated with preterm birth induces monocyte influx into the fetal lung. Previous studies demonstrated that monocytes in the developing lung can mediate lung injury responses that resemble BPD in humans. We hypothesized that chorioamnionitis would induce maturation of immature monocytes in the fetal lung. Groups of three to seven time-mated ewes received saline or 10 mg of endotoxin (Escherichia coli 055:B5) in saline by intra-amniotic injection for intervals from 1 to 14 days before operative delivery at 124 days of gestational age. Monocytic cells from lung tissue were recovered using Percoll gradients. Monocytic cells consistent with macrophages were identified morphologically and by myosin heavy chain class II expression. An increase in macrophages was preceded by induction of granulocyte-macrophage colony-stimulating factor in the lung and subsequent activation of the transcription factor PU.1. The production of IL-6 by monocytes/macrophages in response to endotoxin challenge in vitro increased 7 and 14 days after exposure to intra-amniotic endotoxin. Recombinant TNF-alpha induced IL-6 production by lung monocytic cells exposed to intra-amniotic endotoxin but not in control cells. Monocytic phagocytosis of apoptotic neutrophils also increased 7 and 14 days after exposure to intra-amniotic endotoxin. Intra-amniotic endotoxin induced lung monocytes to develop into functionally mature cells consistent with macrophages. These findings have implications for lung immune responses after exposure to chorioamnionitis.

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

    Science.gov (United States)

    Schwarz, Harald; Schmittner, Maria; Duschl, Albert; Horejs-Hoeck, Jutta

    2014-01-01

    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.

  10. 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 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.

  11. Human endotoxin tolerance is associated with enrichment of the CD14+ CD16+ monocyte subset.

    Science.gov (United States)

    Domínguez-Nieto, Aimée; Zentella, Alejandro; Moreno, José; Ventura, José L; Pedraza, Sigifredo; Velázquez, Juan R

    2015-01-01

    Prior exposure to lipopolysaccharides (LPS) induces a state of cell resistance to subsequent LPS restimulation, known as endotoxin tolerance, mainly by repressing the expression of pro-inflammatory cytokines. We established an endotoxin tolerance model in human monocytes Endotoxin-tolerant cells showed a decrease in IκBα degradation and diminished expression of Tumor necrosis factor (TNF) (both messenger RNA [mRNA] and protein content). The myeloid differentiation factor 88 (MyD88)/MyD88 splice variant (MyD88s) ratio, an indirect way to test the Toll-like receptor 4 (TLR4) MyD88-dependent signaling cascade, did not change in endotoxin-tolerant cells when compared to LPS-stimulated or -unstimulated ones. Remarkably, cell population analysis indicated a significant increase of the CD14+ CD16+ subset only under the endotoxin-tolerant condition. Furthermore, endotoxin-tolerant cells produced higher amounts of C-X-C motif chemokine 10 (CXCL10), a typical MyD88-independent cytokine.

  12. 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.

  13. Chlorogenic acid from honeysuckle improves hepatic lipid dysregulation and modulates hepatic fatty acid composition in rats with chronic endotoxin infusion

    OpenAIRE

    2016-01-01

    Chlorogenic acid as a natural hydroxycinnamic acid has protective effect for liver. Endotoxin induced metabolic disorder, such as lipid dysregulation and hyperlipidemia. In this study, we investigated the effect of chlorogenic acid in rats with chronic endotoxin infusion. The Sprague-Dawley rats with lipid metabolic disorder (LD group) were intraperitoneally injected endotoxin. And the rats of chlorogenic acid-LD group were daily received chlorogenic acid by intragastric administration. In ch...

  14. Understanding the fate of black (pyrogenic) carbon in soil: Preliminary results from a long term field trial

    Science.gov (United States)

    Meredith, Will; Ascough, Philippa; Bird, Michael; Large, David; Shen, Licheng; Snape, Colin

    2014-05-01

    Black carbon (BC, also known as pyrogenic carbon) is an 'inert' form of carbon and has been proposed as a means of long-term carbon sequestration, particularly by amending soils and sediments with BC known as biochar. While there is abundant anecdotal evidence of biochar stability over extended timescales it is essential to gain a greater understanding of the degree and mechanisms of biochar degradation in the environment. This study aims to quantitatively assess the stability of biochar by investigating samples from field degradation trials first buried during 2009 in a tropical soil, and recovered after 12 and 36 month intervals. Catalytic hydropyrolysis (HyPy) is a novel analytical tool for the isolation of BC [1] in which high hydrogen pressure (150 bar) and a sulphided Mo catalyst reductively remove the non-BC fraction of the chars, and so isolate the most stable portion of the biochar, defined as BC(HyPy). This method also allows for the non-BC(HyPy) fraction of a sample, which from charcoal is known to include small ring PAHs (oceans [5], implying that BC cycling could be faster than previously thought [6]. Longer term field trials will be required (analysis of the 36 month samples) to observe systematic changes in the PAH distribution of the non-BCC(HyPy) fractions isolated form aged chars, although natural analogues with a better constrained environmental history may also be useful. [1] Meredith, W. et al., 2012. Geochimica et Cosmochimica Acta 97, 131-147. [2] Ascough, P.L. et al., 2010. Radiocarbon 52, 1336-1350. [3] Lockhart, R.S. et al., 2013. Journal of Analytical and Applied Pyrolysis 104, 640-652. [4] Hockaday, W.C. et al., 2007. Geochimica et Cosmochimica Acta 71, 3432-3445. [5] Jaffe, R. et al., 2013. Science 340, 345-347. [6] Zimmerman, M. et al., 2012. Global Change Biology 18, 3306-3316.

  15. Development of a loop-mediated isothermal amplification method for rapid detection of streptococcal pyrogenic exotoxin B.

    Science.gov (United States)

    Cao, Cuiming; Zhang, Fang; Ji, Mingyu; Pei, Fengyan; Fan, Xiujie; Shen, Hong; Wang, Qingxi; Yang, Weihua; Wang, Yunshan

    2016-07-01

    We developed a visual loop-mediated isothermal amplification (LAMP) technique to detect the streptococcal pyrogenic exotoxin B (speB) gene. Fifteen strains (from American Type Culture Collection or clinical isolates) were used to determine the specificity and sensitivity of the LAMP assay. Clinical samples were collected from 132 patients with suspected Streptococcus pyogenes (S. pyogenes) infection to verify the feasibility of the LAMP assay for detection of the speB gene. By using a set of five primers (a pair of outer primers, a pair of inner primers and one loop primer) targeting the speB gene, the amplification reaction was rapidly performed in a regular water bath under isothermal conditions at 63 °C for approximately 60 min. Only the two S. pyogenes strains showed positive results which were easily observed with the naked eye, and the other strains showed negative results. The detection limit of the LAMP assay was 0.01 ng/μl of template, showing higher sensitivity than conventional PCR (with a detection limit of 1.0 ng/μl). The detection rate of the speB gene in clinical samples was 71.21% and was consistent with the PCR results. The rapid detection of the speB gene by the LAMP assay is highly specific and sensitive, is simple to perform and cost-effective, and is expected to be a new reliable method for the rapid diagnosis of S. pyogenes infection, that is particularly suitable for rural or community hospitals in developing countries.

  16. Plasma endotoxin activity in kangaroos with oral necrobacillosis (lumpy jaw disease) using an automated handheld testing system.

    Science.gov (United States)

    Sotohira, Yukari; Suzuki, Kazuyuki; Sasaki, Haruka; Sano, Tadashi; Tsuchiya, Masakazu; Suzuki, Yohko; Shimamori, Toshio; Tsukano, Kenji; Sato, Ayano; Yokota, Hiroshi; Asakawa, Mitsuhiko

    2016-07-01

    The aim of the present study was to evaluate the reliability and effectiveness of directly determining endotoxin activity in plasma samples from kangaroos with lumpy jaw disease (LJD, n=15) and healthy controls (n=12). Prior to the present study, the ability of the commercially available automated handheld portable test system (PTS(TM)) to detect endotoxin activity in kangaroo plasma was compared with that of the traditional LAL-kinetic turbidimetric (KT) assay. Plasma samples, which were obtained from endotoxin-challenged cattle, were diluted 1:20 in endotoxin-free water and heated to 80°C for 10 min. The performance of the PTS(TM) was not significantly different from that of the traditional LAL-based assay. The data obtained using PTS(TM) correlated with those using KT (r(2)=0.963, PPTS(TM) is applicable as a simplified system to assess endotoxin activity in macropods. In the present study, we demonstrated the diagnostic value of plasma endotoxin activity in kangaroos with systemic inflammation caused by oral necrobacillosis and identified plasma endotoxin activity as a sensitive marker of systemic inflammation in kangaroos with LJD. Based on ROC curves, we proposed a diagnostic cut-off point for endotoxin activity of >0.22 EU/ml for the identification of LJD. Our results indicate that the assessment of plasma endotoxin activity is a promising diagnostic tool for determining the outcome of LJD in captive macropods.

  17. Leakage of Microbial Endotoxin through the Implant-Abutment Interface in Oral Implants: An In Vitro Study

    Directory of Open Access Journals (Sweden)

    Rhoodie Garrana

    2016-01-01

    Full Text Available Background. Endotoxin initiates osteoclastic activity resulting in bone loss. Endotoxin leakage through implant abutment connections negatively influences peri-implant bone levels. Objectives. (i To determine if endotoxin can traverse different implant-abutment connection (IAC designs; (ii to quantify the amount of endotoxins traversing the IAC; (iii to compare the in vitro comportments of different IACs. Materials and Methods. Twenty-seven IACs were inoculated with E. coli endotoxin. Six of the twenty-seven IACs were external connections from one system (Southern Implants and the remaining twenty-one IACs were made up of seven internal IAC types from four different implant companies (Straumann, Ankylos, and Neodent, Southern Implants. Results. Of the 27 IACs tested, all 6 external IACs leaked measurable amounts of endotoxin. Of the remaining 21 internal IACs, 9 IACs did not show measurable leakage whilst the remaining 12 IACs leaked varying amounts. The mean log endotoxin level was significantly higher for the external compared to internal types (p=0.015. Conclusion. Within the parameters of this study, we can conclude that endotoxin leakage is dependent on the design of the IAC. Straumann Synocta, Straumann Cross-fit, and Ankylos displayed the best performances of all IACs tested with undetectable leakage after 7 days. Each of these IACs incorporated a morse-like component in their design. Speculation still exists over the impact of IAC endotoxin leakage on peri-implant tissues in vivo; hence, further investigations are required to further explore this.

  18. Leakage of Microbial Endotoxin through the Implant-Abutment Interface in Oral Implants: An In Vitro Study.

    Science.gov (United States)

    Garrana, Rhoodie; Mohangi, Govindrau; Malo, Paulo; Nobre, Miguel

    2016-01-01

    Background. Endotoxin initiates osteoclastic activity resulting in bone loss. Endotoxin leakage through implant abutment connections negatively influences peri-implant bone levels. Objectives. (i) To determine if endotoxin can traverse different implant-abutment connection (IAC) designs; (ii) to quantify the amount of endotoxins traversing the IAC; (iii) to compare the in vitro comportments of different IACs. Materials and Methods. Twenty-seven IACs were inoculated with E. coli endotoxin. Six of the twenty-seven IACs were external connections from one system (Southern Implants) and the remaining twenty-one IACs were made up of seven internal IAC types from four different implant companies (Straumann, Ankylos, and Neodent, Southern Implants). Results. Of the 27 IACs tested, all 6 external IACs leaked measurable amounts of endotoxin. Of the remaining 21 internal IACs, 9 IACs did not show measurable leakage whilst the remaining 12 IACs leaked varying amounts. The mean log endotoxin level was significantly higher for the external compared to internal types (p = 0.015). Conclusion. Within the parameters of this study, we can conclude that endotoxin leakage is dependent on the design of the IAC. Straumann Synocta, Straumann Cross-fit, and Ankylos displayed the best performances of all IACs tested with undetectable leakage after 7 days. Each of these IACs incorporated a morse-like component in their design. Speculation still exists over the impact of IAC endotoxin leakage on peri-implant tissues in vivo; hence, further investigations are required to further explore this.

  19. The role of endotoxin in the pathogenesis of gastric mucosal damage in cirrhotic rats with portal hypertensive gastropathy

    Institute of Scientific and Technical Information of China (English)

    ChengLan; XiaoningSun; LiweiDong; BailiHuang; SuYuan; KeliWu

    2011-01-01

    Objective:To study the role and the mechanism of endotoxin in the pathogenesis of gastric mucosa during portal vein hypertension gastrography (PHG) in the rats with cirrhosis. Methods:Rat model for PHG was established by injection of tetrachloride. The animals were injected with endotoxin i.p. at 3 mg/kg and endotoxin antagonist BPI21 i.v. at 2.0 mg/kg. The plasma level of endotoxin as well as the gastric mucosal level of tumor necrosis factor alpha (TNF-α) was measured with azobenzene and ELISA respectively. Furthermore, the pathological changes of the gastric mucosa were studied with HE stainning. Results:In rats with PHG, increased endotoxin and TNF-αas well as the gastric pathological lesion were observed. Injection of endotoxin remarkably increased plasma level of endotoxin as well as the gastric mucosal level of tumor necrosis TNF-αand induced more serious gastric lesion. Animals injected with endotoxin antagonist BPI21 showed improved gastric mucosal lesion, accompanied by the declining TNF-αlevel. Conclusions:Our results suggestes that endotoxin may play a pathogenetic role in PHG by inducing the expression of TNF-α.

  20. 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.

  1. Rifaximin for the prevention of spontaneous bacterial peritonitis

    Institute of Scientific and Technical Information of China (English)

    Georgios N Kalambokis; Athanasia Mouzaki; Maria Rodi; Epameinondas V Tsianos

    2012-01-01

    According to a review article by Biecker etal published in a previous issue of World Journal of Gastroenterology in March 2011,intestinal decontamination with norfloxacin remains the mainstay of primary prophylaxis of spontaneous bacterial peritonitis (SBP) at the expense of development of quinolone-resistant bacteria after long-term use.In our research,the administration of a 4-wk regimen with rifaximin 1200 mg/d reduced significantly the ascitic neutrophil count in cirrhotic patients with sterile ascites in line with a significant decrease in plasma endotoxin levels.Our observations concur with recent findings,showing a significantly reduced 5-year probability of SBP in cirrhotic patients taking rifaximin.

  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

    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.

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

    Science.gov (United States)

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

    2007-04-01

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

  5. Bacterial gastroenteritis

    Science.gov (United States)

    Bacterial gastroenteritis is present when bacteria cause an infection of the stomach and intestines ... has not been treated Many different types of bacteria can cause ... Campylobacter jejuni E coli Salmonella Shigella Staphylococcus ...

  6. Cardiorenal Syndrome Type 5 in Sepsis: Role of Endotoxin in Cell Death Pathways and Inflammation

    Directory of Open Access Journals (Sweden)

    Grazia Maria Virzì

    2016-12-01

    Full Text Available Background/Aims: Cardiorenal Syndrome Type 5 (CRS Type 5 is characterized by concomitant cardiac and renal dysfunction in the setting of different systemic disorders, such as sepsis. In this study, we investigated the possible relationship between endotoxin levels, renal cell death and inflammation in septic patients with CRS Type 5. Methods: We enrolled 11 patients with CRS Type 5. CRS Type 5 was defined according to the current classification system. AKI was defined by Acute Kidney Injury Network (AKIN criteria. Acute cardiac dysfunction was documented by echocardiography as acute left and/or right ventricular dysfunction leading to decreased ejection fraction. Endotoxin activity was measured by the Endotoxin Activity Assay (EAA. Plasma from CRS Type 5 patients was incubated with renal tubular cells (RTCs and cell death levels were evaluated. Plasma cytokines levels were measured as well. Results: Accordingly to EAA levels, patients were divided into two groups: 45.4% of patients had low endotoxin activity level (negative EAA, while 54.5% of patients showed high endotoxin activity (positive EAA. RTCs incubated with plasma from EAA positive patients showed significantly higher apoptosis levels and higher caspase-3 activation compared to cells incubated with plasma from EAA negative patients, and a significant positive correlation was observed between EAA levels and RTC apoptosis levels. Furthermore, IL-6 and IFN-γ levels were significantly higher in CRS Type 5 patients with positive EAA. Conclusion: Our data suggest a possible relationship between endotoxin levels and renal cell death in septic patients with CRS Type 5. Furthermore, this study highlights the presence of renal apoptosis, the immune deregulation and the strong inflammation in CRS Type 5 patients, especially in those with high endotoxin activity.

  7. Albumin synthesis in humans increases immediately following the administration of endotoxin.

    Science.gov (United States)

    Barle, Hans; Januszkiewicz, Anna; Hållström, Lars; Essén, Pia; McNurlan, Margaret A; Garlick, Peter J; Wernerman, Jan

    2002-11-01

    In order to investigate the immediate (i.e. within 3 h) response of albumin synthesis to the administration of endotoxin, as a model of a moderate and well controlled catabolic insult, two measurements employing L-[(2)H(5)]phenylalanine were performed in 16 volunteers. One group ( n =8) received an intravenous injection of endotoxin (4 ng/kg; lot EC-6) immediately after the first measurement of albumin synthesis, whereas the other group received saline. A second measurement was initiated 1 h later. In the endotoxin group, the fractional synthesis rate of albumin was 6.9+/-0.6%/day (mean+/-S.D.) in the first measurement. In the second measurement, a significant increase was observed (9.6+/-1.2%/day; P <0.001). The corresponding values in the control group were were 6.6+/-0.6%/day and 7.0+/-0.6%/day respectively (not significant compared with first measurement and P <0.001 compared with the second measurement in the endotoxin group). The absolute synthesis rates of albumin were 148+/-35 and 201+/-49 mg x kg(-1) x day(-1) before and after endotoxin ( P <0.01). In the control group, the corresponding values were 131+/-21 and 132+/-20 mg x kg(-1) x day(-1) (not significant compared with the first measurement and P <0.01 compared with the second measurement in the endotoxin group). In conclusion, these results indicate that albumin synthesis increases in the very early phase after a catabolic insult, as represented by the administration of endotoxin.

  8. 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.

  9. Inhibition of adenosine deaminase attenuates endotoxin-induced release of cytokines in vivo in rats.

    Science.gov (United States)

    Tofovic, S P; Zacharia, L; Carcillo, J A; Jackson, E K

    2001-09-01

    The purpose of this study was to investigate in vivo the effects of modulating the adenosine system on endotoxin-induced release of cytokines and changes in heart performance and neurohumoral status in early, profound endotoxemia in rats. Time/pressure variables of heart performance and blood pressure were recorded continuously, and plasma levels of tumor necrosis factor alpha (TNFalpha), interleukin 1-beta (IL-1beta), plasma renin activity (PRA), and catecholamines were determined before and 90 min after administration of endotoxin (30 mg/kg of lipopolysaccharide, i.v.). Erythro-9[2-hydroxyl-3-nonyl] adenine (EHNA; an adenosine deaminase inhibitor) had no effects on measured time-pressure variables of heart performance under baseline conditions and during endotoxemia, yet significantly attenuated endotoxin-induced release of cytokines and PRA. Pretreatment with the non-selective adenosine receptor antagonist DPSPX not only prevented the effects of EHNA but also increased the basal release of cytokines and augmented PRA. At baseline, caffeine (a non-selective adenosine receptor antagonist) increased HR, +dP/dtmax, heart rate x ventricular pressure product (HR x VPSP) and +dP/dtmax normalized by pressure (+dP/dtmax/VPSP), and these changes persisted during endotoxemia. Caffeine attenuated endotoxin-induced release of cytokines and augmented endotoxin-induced increases in plasma catecholamines and PRA. Pretreatment with propranolol abolished the effects of caffeine on heart performance and neurohumoral activation during the early phase of endotoxemia. 6N-cyclopentyladenosine (CPA; selective A1 adenosine receptor agonist) induced bradicardia and negative inotropic effects, reduced work load (i.e., decreased HR, VPSP, +dP/dtmax, +dP/dtmax/VPSP and HR x VPSP) and inhibited endotoxin-induced tachycardia and renin release. CGS 21680 (selective A2A adenosine receptor agonist) decreased blood pressure under basal condition but did not potentiate decreases in blood pressure

  10. The global pyrogenic carbon cycle and its impact on the level of atmospheric CO2 over past and future centuries.

    Science.gov (United States)

    Landry, Jean-Sébastien; Matthews, H Damon

    2016-12-19

    The incomplete combustion of vegetation and dead organic matter by landscape fires creates recalcitrant pyrogenic carbon (PyC), which could be consequential for the global carbon budget if changes in fire regime, climate, and atmospheric CO2 were to substantially affect gains and losses of PyC on land and in oceans. Here, we included global PyC cycling in a coupled climate-carbon model to assess the role of PyC in historical and future simulations, accounting for uncertainties through five sets of parameter estimates. We obtained year-2000 global stocks of (Central estimate, likely uncertainty range in parentheses) 86 (11-154), 47 (2-64), and 1129 (90-5892) Pg C for terrestrial residual PyC (RPyC), marine dissolved PyC, and marine particulate PyC, respectively. PyC cycling decreased atmospheric CO2 only slightly between 1751 and 2000 (by 0.8 Pg C for the Central estimate) as PyC-related fluxes changed little over the period. For 2000 to 2300, we combined Representative Concentration Pathways (RCPs) 4.5 and 8.5 with stable or continuously increasing future fire frequencies. For the increasing future fire regime, the production of new RPyC generally outpaced the warming-induced accelerated loss of existing RPyC, so that PyC cycling decreased atmospheric CO2 between 2000 and 2300 for most estimates (by 4-8 Pg C for Central). For the stable fire regime, however, PyC cycling usually increased atmospheric CO2 (by 1-9 Pg C for Central), and only the most extreme choice of parameters maximizing PyC production and minimizing PyC decomposition led to atmospheric CO2 decreases under RCPs 4.5 and 8.5 (by 5-8 Pg C). Our results suggest that PyC cycling will likely reduce the future increase in atmospheric CO2 if landscape fires become much more frequent; however, in the absence of a substantial increase in fire frequency, PyC cycling might contribute to, rather than mitigate, the future increase in atmospheric CO2 .

  11. Linking charring temperature and wood source to the structure and degradation rates of pyrogenic organic matter in soil

    Science.gov (United States)

    Hatton, P.; Dastmalchi, K.; Chatterjee, S.; Auclerc, A.; Le Moine, J.; Filley, T. R.; Nadelhoffer, K. J.; Stark, R.; Bird, J. A.

    2013-12-01

    Fire is a major controller of forest C cycling by releasing CO2 to the atmosphere and by contributing pyrogenic organic matter (PyOM or biochar) to soils. Recent studies have shown that much of fire-derived PyOM may turn over in soils at century time scales. Two likely controllers of the chemical structure of PyOM and its resulting decay rate are pyrolysis temperature and the source biomass. However, we know little of how these two factors determine the chemical structure and bioreactivity of the resulting PyOM. To gain further insight into controls on the structure and fate of PyOM, we examined two species of dual-labeled (13C/15N), wood-based PyOM (Pinus banksiana and Acer rubrum) made with 5 pyrolysis temperatures (0, 200, 300, 450, 600 °C) using solid state nuclear magnetic resonance, isotopic and elemental composition (C, H, O, and N), and differential scanning calorimetry. In addition, we are investigating the fate of a subset of these PyOM materials applied to forest soils in a long-term field study located at the University of Michigan Biological Station in Pellston, MI, USA. We will present data of the loss of PyOM C as CO2 and DOC during the first year in situ. We found complementary lines of evidence for a facile removal of cellulose and hemicellulose and a progressive alteration of nitrogenous moieties across the charring gradient for wood-derived PyOM of both tree species as temperature was increased from 0 to 600 °C. Our NMR results show a significant species by pyrolysis temperature interaction on PyOM chemical structure with considerably less condensation for Acer- than Pinus-derived PyOM at 300 °C. In the first year after addition to soil, Acer-derived PyOM pyrolyzed at 450 °C mineralized faster than Pinus-derived PyOM pyrolyzed at 450 °C. Increasing pyrolysis temperatures for Pinus-derived PyOM also resulted in slower CO2 mineralization rates during the first year of field decay. These results relate pyrolysis temperature to the resulting Py

  12. Demonstration of interleukin 1 activity in apparently homogeneous specimens of the pI 5 form of rabbit endogenous pyrogen.

    Science.gov (United States)

    Hanson, D F; Murphy, P A

    1984-08-01

    Rabbit mononuclear cells from oil-induced peritoneal exudates were purified by centrifugation on Percoll gradients, suspended in tissue culture medium, and stimulated with opsonized Staphylococcus epidermidis. The supernatants from these macrophages caused fever when injected intravenously into rabbits (endogenous pyrogen [EP] activity). The EP activity was contained in two protein fractions, with pIs of 7.3 and ca. 5.0. The same fractions caused mouse thymocytes to incorporate tritiated thymidine when incubated in vitro with small quantities of phytohemagglutinin (interleukin 1 [IL-1] activity). The pI 5.0 form of EP was purified to apparent homogeneity by sequential use of ammonium sulfate precipitation, gel filtration, ion-exchange chromatography, hydrophobic chromatography, and high-resolution isoelectric focusing. EP and IL-1 activities were not separable by any of these procedures. Active fractions from isoelectric focusing were analyzed by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Only one band was visible as judged by a silver staining method, and IL-1 activity could be recovered by renaturing eluates from the same region of sodium dodecyl sulfate gels run in parallel. An estimate of specific activity was made by comparing the intensity of stained bands of EP with the intensity of bands containing known quantities of lysozyme or RNase. By this criterion, the specific activity of purified pI 5 EP was between 17,000 and 58,000 degrees C U/mg of protein, and the specific activity in terms of IL-1 was between 59 million and 360 million U per mg of protein. These observations suggest that both EP and IL-1 activities can be expressed by a single molecular species. The implications of this coincidence are discussed. It was also shown that highly purified pI 5 EP obtained from macrophages stimulated in the presence of 14C-labeled amino acids contained significant 14C radioactivity. This suggests that the pI 5.0 EP, like the pI 7

  13. 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.

  14. 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.

  15. 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

  16. 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.

  17. 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.

  18. Effects of hypothermia on blood endogenous endotoxin levels during cardiopulmonary bypass.

    Science.gov (United States)

    Gerçekoglu, H; Tarim, O; Agar, I; Korukçu, A; Karabulut, H; Soydemir, H; Sokullu, O; Toklu, H; Johansson, C B; Yigiter, B; Kopman, E

    1997-01-01

    Endotoxin activates white blood cells and complement and produces a spectrum of clinical syndromes ranging from fever to septic shock. Although production of endogenous endotoxemia during cardiopulmonary bypass (CPB) has recently been reported, the role of hypothermia on endotoxemia is not clear. In this study, we evaluated the effects of moderate (24-28 degrees C) and mild (32-34 degrees C) hypothermia on blood endotoxin levels. The study population consisted of 20 patients who underwent coronary artery bypass grafting (CABG) with CPB. Moderate systemic hypothermia was applied during aortic cross-clamping in ten patients (group 1) and mild hypothermia in the remaining ten patients (group 2). The mean rectal temperatures were 26.8 +/- 1.2 degrees C in group 1 and 33.8 +/- 0.8 degrees C in group 2. The blood samples for endotoxin level measurements were obtained before CPB, during aortic cross-clamping, immediately after the release of the cross-clamp, 20 minutes after the release of the cross-clamp, after CPB, and 2 hours postoperatively. There were no endotoxins in any of the samples before CPB, but it was detected after CPB in both groups. The endotoxin levels were significantly higher in group 1 than in group 2. The present study suggests that when hypothermia is the technique of choice, the deleterious effects of endotoxemia on patients with comorbidity must be considered.

  19. 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... From Tolerances § 180.1108 Delta endotoxin of Bacillus thuringiensis variety San Diego encapsulated... of Bacillus thuringiensis variety San Diego encapsulated into killed Pseudomonas fluorescens...

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

    NARCIS (Netherlands)

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

    1997-01-01

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

  1. 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.

  2. Endotoxin deposits on the inner surfaces of closed-face cassettes during bioaerosol sampling: a field investigation at composting facilities.

    Science.gov (United States)

    Duquenne, Philippe; Simon, Xavier; Demange, Valérie; Harper, Martin; Wild, Pascal

    2015-05-01

    A set of 270 bioaerosol samples was taken from 15 composting facilities using polystyrene closed-face filter cassettes (CFCs). The objective was to measure the quantity of endotoxin deposits on the inner surfaces of the cassettes (sometimes referred to as 'wall deposits'). The results show that endotoxins are deposited on the inner surfaces of the CFCs through sampling and/or handling of samples. The quantity of endotoxins measured on inner surfaces range between 0.05 (the limit of detection of the method) and 3100 endotoxin units per cassette. The deposits can represent a large and variable percentage of the endotoxins sampled. More than a third of the samples presented a percentage of inner surface deposits >40% of the total quantity of endotoxins collected (filter + inner surfaces). Omitting these inner surface deposits in the analytical process lead to measurement errors relative to sampling all particles entering the CFC sampler, corresponding to a developing consensus on matching the inhalable particulate sampling convention. The result would be underestimated exposures and could affect the decision as to whether or not a result is acceptable in comparison to airborne concentration limits defined in terms of the inhalability convention. The results of this study suggest including the endotoxins deposited on the inner surfaces of CFCs during analysis. Further researches are necessary to investigate endotoxin deposits on the inner cassette surfaces in other working sectors.

  3. Amide-mediated hydrogen bonding at organic crystal/water interfaces enables selective endotoxin binding with picomolar affinity.

    Science.gov (United States)

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

    2013-05-22

    Since the discovery of endotoxins as the primary toxic component of Gram-negative bacteria, researchers have pursued the quest for molecules that detect, neutralize, and remove endotoxins. Selective removal of endotoxins is particularly challenging for protein solutions and, to this day, no general method is available. Here, we report that crystals of the purine-derived compound allantoin selectively adsorb endotoxins with picomolar affinity through amide-mediated hydrogen bonding in aqueous solutions. Atom force microscopy and chemical inhibition experiments indicate that endotoxin adsorption is largely independent from hydrophobic and ionic interactions with allantoin crystals and is mediated by hydrogen bonding with amide groups at flat crystal surfaces. The small size (500 nm) and large specific surface area of allantoin crystals results in a very high endotoxin-binding capacity (3 × 10(7) EU/g) which compares favorably with known endotoxin-binding materials. These results provide a proof-of-concept for hydrogen bond-based molecular recognition processes in aqueous solutions and establish a practical method for removing endotoxins from protein solutions.

  4. 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.

  5. 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

  6. Induction of generalized Shwartzman reaction (GSR) in irradiated rabbits by a single injection of endotoxin

    Energy Technology Data Exchange (ETDEWEB)

    Wronowski, T.; Uchanska-Dudzinska, B.; Teisseyre, E.; Kopec, M.

    1976-04-01

    Rabbits were prepared to GSR by either a single dose 30 ..mu..g/kg of Salmonella enteritidis endotoxin or by a single whole-body exposure to 850 R of x ray. After 24 hrs provocative dose of endotoxin 50 ..mu..g/kg was injected. Histological examinations of kidneys revealed that GSR occurred with equal frequency in both groups of rabbits but was more severe in the animals prepared by irradiation. Protamine gelation test detected soluble fibrin complexes in plasma in about 50% of rabbits 24 hrs after either of the preparatory procedures and virtually in 100% after the provocative endotoxin dose. During the preparatory phase of GSR the irradiation induced a profound decrease in blood lymphocyte count and in the weight of thymus and spleen. It is postulated that early postexposure lysis of lymphatic cells participates in mechanisms leading to thrombotic phenomena after irradiation.

  7. 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.

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

    DEFF Research Database (Denmark)

    Hojman, Pernille; Taudorf, Sarah; Lundby, Carsten;

    2009-01-01

    Recent studies have shown that erythropoietin (EPO) offers protection against ischemia, hemorrhagic shock and systemic inflammation in many tissues and it has been suggested that EPO has anti-inflammatory effects. With the aim of investigating the potential acute anti-inflammatory effects of EPO...... receiving either (1) LPS alone, (2) EPO alone (15,000 IE of rHuEPO) or (3) EPO and LPS. Endotoxin administration alone induced a 3-, 12- and 5-fold increase in plasma concentrations of TNF-alpha, IL-6 and IL-10, respectively, 3h after LPS challenge. When EPO was given prior to a bolus injection...... with endotoxin, the levels of TNF-alpha and IL-6 were enhanced by 5- and 40-fold, respectively, whereas the endotoxin-induced increase in IL-10 response was not influenced by EPO. In contrast to our hypothesis, we find that EPO augments the acute inflammatory effect....

  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 Contamination and Correlation with Other Water Quality Parameters of Groundwater from Self-Contained Wells in Beijing].

    Science.gov (United States)

    Zhang, Can; Liu, Wen-jun; Ao, Lu; Shi, Yun; An, Dai-zhi; Liu, Zhi-ping

    2015-12-01

    A survey of endotoxin activity in groundwater from 14 self-contained wells in PLA units stationed in Beijing was conducted by the kinetic-turbid assay of Tachypleus Amebocyte Lysate (TAL). Bacteriological parameters, including total cell counts detected by flow cytometry, heterotrophic plate counts (HPC), standard plate counts and total coliforms were analyzed. Additionally, suspended particles, turbidity, dissolved organic carbon (DOC), and UV₂₅₄ were investigated. Total endotoxin activities ranged from 0. 15 to 13.20 EU · mL⁻¹, free endotoxin activities ranged from 0.10 to 5.29 EU · mL⁻¹ and bound endotoxin activities ranged from 0.01 to 8.60 EU · mL⁻¹. Most of the endotoxins in heavily contaminated groundwater existed as bound endotoxins. As for total endotoxins, the sequence of correlation coefficients with other parameters was total cell counts (r = 0.88 ) > HPC (r = 0.79) > DOC (r = 0.77) > UV₂₅₄ (r = 0.57) > total coliforms (r = 0.50) > standard plate counts (r = 0.49) = turbidity (r = 0. 49) > total particles (r = 0.41). The sequence of correlations of the bound endotoxins with other parameters was total cell counts (r = 0.81) > HPC (r = 0.66) > total coliforms (r = 0.65) > turbidity (r = 0.62) > total particles (r = 0.58) > standard plate counts (r = 0.22). Free endotoxins were correlated with DOC and UV₂₅₄, r = 0.58 and 0.26, respectively. Result showed free endotoxins had a higher correlation with DOC, and a lower correlation with UV₂₅₄.

  11. Comparative studies of endotoxin uptake by isolated rat Kupffer and peritoneal cells.

    Science.gov (United States)

    Fox, E S; Thomas, P; Broitman, S A

    1987-12-01

    The process of uptake of endotoxin by cells of the reticuloendothelial system is of current interest. Rabbit peritoneal macrophages have been used to study macrophage-endotoxin interactions and have suggested a receptor-mediated process. It is generally believed that the site of in vivo endotoxin clearance is the liver and that this clearance involves the Kupffer cell population. In the current report, the uptake characteristics of iodine-125-labeled Salmonella minnesota lipopolysaccharide (LPS) were compared in both isolated rat Kupffer cells and elicited rat peritoneal cells. Both types of cells were isolated from male Sprague-Dawley rats fed a semisynthetic AIN-76 5% saturated-fat diet either by peritoneal lavage for peritoneal cells or by collagenase perfusion followed by purification on a 17.5% metrizamide gradient for Kupffer cells. Hot phenol water-extracted S. minnesota LPS was labeled with iodine by the chloramine-T method following a reaction with methyl-p-hydroxybenzimidate. The in vitro uptake of [125I]LPS by Kupffer cells was unsaturable up to concentrations of 33.33 micrograms/ml, while peritoneal cells became saturated at between 16.67 and 25 micrograms of LPS per ml. Uptake by both types of cells could be inhibited by a 10-fold excess of unlabeled LPS. Kinetic experiments demonstrated that Kupffer cells were unsaturable after 60 min of incubation, while peritoneal cells were saturable after 40 min of incubation. Pretreatment with 75 mM colchicine inhibited uptake by peritoneal cells but not Kupffer cells, while pretreatment with 12 mM 2-deoxyglucose inhibited uptake by Kupffer cells but not peritoneal cells. These results are consistent with a process of receptor-mediated endocytosis for peritoneal cells, while Kupffer cells may internalize endotoxins by absorptive pinocytosis. These results suggest that studies of peritoneal cell-endotoxin interactions do not accurately describe the physiologic process within the liver, the major site for the

  12. Biphasic fermentation is an efficient strategy for the overproduction of δ-endotoxin from Bacillus thuringiensis.

    Science.gov (United States)

    Jisha, Veloorvalappil Narayanan; Smitha, Robinson Babysarojam; Priji, Prakasan; Sajith, Sreedharan; Benjamin, Sailas

    2015-02-01

    This study illustrates a biphasic solid-state fermentation (SSF) strategy for the overproduction of δ-endotoxin from Bacillus thuringiensis subsp. kurstaki (Btk) and also purification of δ-endotoxin from the solid-fermented medium. The fermentation strategy had two phases (biphasic); i.e., the first short phase was semisolid state (12 h), and the remaining long phase was strict SSF. To achieve the biphasic SSF, after 12 h (150 rpm, 37 °C) fermentation of the medium [Luria-Bertani (LB) supplemented with 30 % (w/v) raw soybean flour (phase I)], the supernatant in it was completely centrifuged out (1,000 × g, 10 min) aseptically for harvesting the extracellular enzymes as by-product. The resultant wet solid matter without free-flowing liquid but with embedded Btk was incubated 60 h more (phase II) for enhancing δ-endotoxin production at static condition (37 °C). Coupled with this, δ-endotoxin was purified by the modified phase separation method, and its purity was physically confirmed by both staining and microscopic techniques. The maximum δ-endotoxin yield from solid medium (48 h) was 15.8 mg/mL (recovery was 55-59 %) LB-equivalent, while that of LB control (recovery was 95 %) was only 0.43 mg/mL (72 h), i.e., thus, in comparison, 36.74-fold more yield in solid medium obtained by 24 h less gestation period. The purified crystal proteins showed apparent molecular weights (MWs) of 45, 35, and 6 kDa on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Briefly, this unique study physically demonstrates how Btk δ-endotoxin is purified (95-99 % purity) from solid-fermented matter for the first time, coupled with its overproduction at the expense of only 21.5 % higher production cost.

  13. 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.

  14. ISOLATION OF ENDOTOXIN-SPECIFIC ANTIBODIES BY SELECTION OF AN SINGLE CHAIN PHAGE ANTIBODY LIBRARY

    Institute of Scientific and Technical Information of China (English)

    陈鸣; 俞丽丽; 张雪; 府伟灵

    2002-01-01

    Objective: To isolate murine anti endotoxin single chain phage antibody from a constructed library. Methods: Total RNA was firstly extracted from murine splenic cells and mRNA was reverse-transcribed into cDNA. Then the designed primers were used to amplify the variable region genes of the heavy and light chain (VH, VL) with polymerase chain reaction. The linker was used to assemble the VH and VL into ScFv, and the NotI and SfiI restriction enzymes were used to digest the ScFv in order to ligate into the pCANTAB5E phagemid vector that was already digested with the same restriction enzymes. The ligated vector was then introduced into competent E.coli TG1 cells to construct a single-chain phage antibody library. After rescued with M13KO7 helper phage, recombinant phages displaying ScFv fragments were harvested from the supernatant and selected with endotoxin. The enriched positive clones were reinfected into TG1 cells. Finally, 190 clones were randomly selected to detect the anti endotoxin antibody with indirect ELISA. Results: The titer of anti endotoxin in murine sera was 1:12,800. The concentration of total RNA was 12.38 μg/ml. 1.9×107 clones were obtained after transformed into TG1. 3×104 colonies were gotten after one round panning. Two positive colonies were confirmed with indirect ELISA among 190 randomly selected colonies. Conclusion: A 1.9×107 murine anti endotoxin single chain phage antibody library was successfully constructed. Two anti endotoxin antibodies were obtained from the library.

  15. 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...

  16. 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

  17. 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...

  18. Experimental study of 『PERSICAE SEMEN』 on the blood injected by Endotoxin in rats

    OpenAIRE

    Chang-Keun; Kyeong-Sun Soh; Chan-Gil Jeong

    2005-01-01

    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 sign...

  19. Predictors of coarse particulate matter and associated endotoxin concentrations in residential environments

    Science.gov (United States)

    Bari, Md. Aynul; MacNeill, Morgan; Kindzierski, Warren B.; Wallace, Lance; Héroux, Marie-Ève; Wheeler, Amanda J.

    2014-08-01

    Exposure to coarse particulate matter (PM), i.e., particles with an aerodynamic diameter between 2.5 and 10 μm (PM10-2.5), is of increasing interest due to the potential for health effects including asthma, allergy and respiratory symptoms. Limited information is available on indoor and outdoor coarse PM and associated endotoxin exposures. Seven consecutive 24-h samples of indoor and outdoor coarse PM were collected during winter and summer 2010 using Harvard Coarse Impactors in a total of 74 Edmonton homes where no reported smoking took place. Coarse PM filters were subsequently analyzed for endotoxin content. Data were also collected on indoor and outdoor temperature, relative humidity, air exchange rate, housing characteristics and occupants' activities. During winter, outdoor concentrations of coarse PM (median = 6.7 μg/m3, interquartile range, IQR = 3.4-12 μg/m3) were found to be higher than indoor concentrations (median 3.4 μg/m3, IQR = 1.6-5.7 μg/m3); while summer levels of indoor and outdoor concentrations were similar (median 4.5 μg/m3, IQR = 2.3-6.8 μg/m3, and median 4.7 μg/m3, IQR = 2.1-7.9 μg/m3, respectively). Similar predictors were identified for indoor coarse PM in both seasons and included corresponding outdoor coarse PM concentrations, whether vacuuming, sweeping or dusting was performed during the sampling period, and number of occupants in the home. Winter indoor coarse PM predictors also included the number of dogs and indoor endotoxin concentrations. Summer median endotoxin concentrations (indoor: 0.41 EU/m3, outdoor: 0.64 EU/m3) were 4-fold higher than winter concentrations (indoor: 0.12 EU/m3, outdoor: 0.16 EU/m3). Other than outdoor endotoxin concentrations, indoor endotoxin concentration predictors for both seasons were different. Winter endotoxin predictors also included presence of furry pets and whether the vacuum had a high efficiency particulate air (HEPA) filter. Summer endotoxin predictors were problems with mice in the

  20. Tetrahydrobiopterin corrects Escherichia coli endotoxin-induced endothelial dysfunction.

    Science.gov (United States)

    Mittermayer, Friedrich; Pleiner, Johannes; Schaller, Georg; Zorn, Stefan; Namiranian, Khodadad; Kapiotis, Stylianos; Bartel, Gregor; Wolfrum, Mathias; Brügel, Mathias; Thiery, Joachim; Macallister, Raymond J; Wolzt, Michael

    2005-10-01

    Acute inflammation causes endothelial dysfunction, which is partly mediated by oxidant stress and inactivation of nitric oxide. The contribution of depletion of tetrahydrobiopterin (BH(4)), the cofactor required for nitric oxide generation, is unclear. In this randomized, double-blind, three-way crossover study, forearm blood flow (FBF) responses to ACh and glyceryltrinitrate (GTN) were measured before and 3.5 h after infusion of Escherichia coli endotoxin (LPS, 20 IU/kg iv) in eight healthy men. The effect of intra-arterial BH(4) (500 microg/min), placebo, or vitamin C (24 mg/min) was studied on separate days 3.5 h after LPS infusion. In addition, human umbilical vein endothelial cells were incubated for 24 h with vitamin C and LPS. ACh and GTN caused dose-dependent forearm vasodilation. The FBF response to ACh, which was decreased by 23 +/- 17% (P < 0.05) by LPS infusion, was restored to baseline reactivity by BH(4) and vitamin C. FBF responses to GTN were not affected by BH(4) or vitamin C. LPS increased leukocyte count, high-sensitivity C-reactive protein, IL-6, IL-1beta, IFN-gamma, monocyte chemoattractant protein-1, pulse rate, and body temperature and decreased platelet count and vitamin C concentration. Vitamin C increased forearm plasma concentration of BH(4) by 32% (P < 0.02). Incubation with LPS and vitamin C, but not LPS alone, increased intracellular BH(4) concentration in human umbilical vein endothelial cells. Impaired endothelial function during acute inflammation can be restored by BH(4) or vitamin C. Vitamin C may exert some of its salutary effects by increasing BH(4) concentration.

  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. EFFECT OF RADIX SALVIAE MILTIORRHIZAE ON THE PROCOAGULANT AND FIBRINOLYTIC ACTIVITIES INDUCED BY ENDOTOXIN-TREATED BOVINE ENDOTHELIAL CELLS

    Institute of Scientific and Technical Information of China (English)

    沈传陆; 张彩英; 王振义

    1992-01-01

    Effect of endotoxin, and endotoxin plus Radix Salviae Miltiorrhizae (RSM) on bovine aortic endothelial cells (BAECs) for the production of tissue factor (TF), plasminogen activator (PA) and plasminogen activator inhibitor (PAI) activities as well as prostacyclin (PGI2) content were studied. Stimulation of BAECs with endotoxin (1 μg/ ml) increased the expression of cell surface TF activity, the secretions of PAI activity and PGI2 content. However, PA activity in endotoxin-treated groups didn’t change significantly in comparison with that of the control. RSM could stimulate the secretion of PA activity and prevent the increase in TF and PAI activities from endotoxin-treated BAECs. However, there was no significant difference in PGI2 production between endotoxin-and endotoxin+RSM-treated BAECs. These results indicated that increased procoagulant function and decreased fibrinolytic activity of endothelial cells (ECs) might be one of the mechanisms of DIC in gram-negative septicemia and RSM would be a very important thug in the prevention and the treatment of DIC in gram-negative sepsis.

  3. [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.

  4. Decreased beating rate variability of spontaneously contracting cardiomyocytes after co-incubation with endotoxin.

    Science.gov (United States)

    Schmidt, Hendrik; Saworski, Jana; Werdan, Karl; Müller-Werdan, Ursula

    2007-01-01

    Decreased heart rate variability (HRV) in critically ill patients indicates a poor prognosis. In heart failure patients, there is an elevated sympathetic tone, reflected by a dominance of sympathetic parameters in HRV, whereas in critically ill patients sympathetic and parasympathetic modulation of heart rate is attenuated despite increased catecholamine blood levels. Thus, autonomic dysfunction in the critically ill cannot be causally related to an impairment at the level of neural transmission, but may be due to a derangement of signal transduction at the effector cell level. On the basis of our working hypothesis that endotoxin may be involved in this blunting of effector cell response to nerval input, we studied the spontaneous beating of cardiomyocytes under the influence of endotoxin. Applying the clinically established indices of HRV to the analysis of beating rate variability (BRV) of neonatal rat cardiomyocytes in serum-free medium, a narrowing of their BRV by endotoxin is demonstrated. We propose that the narrowing of HRV in critically ill patients does not only reflect the altered input from the central or peripheral neurons, but rather a remodeling of the cardiac pacemaker cells by endotoxin and inflammatory mediators.

  5. 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 (PPTS 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.

  6. 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

  7. 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

  8. 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

  9. 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

  10. Differential acute phase immune responses by Angus and Romosinuano steers following an endotoxin challenge

    Science.gov (United States)

    Our primary objective was to evaluate potential genetic differences between two diverse Bos taurus breeds (Angus (AG) and Romosinuano (RO)) in response to an endotoxin. The RO is a tropically adaptive Bos taurus breed developed in the Sinú valley of northern Colombia. Eighteen steers (n = 9 steers/b...

  11. Concentrations of airborne endotoxin and microorganisms at a 10,000 cow open-freestall dairy

    Science.gov (United States)

    Confined animal production systems produce elevated bioaerosol concentrations, which are a potential respiratory health risk to individuals on site and downwind. In this study, airborne endotoxin and microorganisms were collected during the spring, summer, and fall at a large open-freestall dairy i...

  12. Yeast cell wall supplementation alters the metabolic responses of crossbred heifers to an endotoxin challenge

    Science.gov (United States)

    This study examined the effect of feeding yeast cell wall (YCW) products on the metabolic responses of newly-received heifers to endotoxin challenge. Heifers (n = 24; 219 ± 2.4 kg) were separated into treatment groups receiving a Control diet (n = 8), YCW-A (2.5 grams/heifer/d; n = 8) or YCW-C (2.5 ...

  13. Evaluation of robot automated chromogenic substrate LAL endotoxin assay method for pharmaceutical products testing.

    Science.gov (United States)

    Tsuji, K; Martin, P A

    1985-01-01

    The robot automated chromogenic substrate LAL assay method was evaluated for endotoxin testing using three lots each of 12 pharmaceutical products. As many as 216 assays, including automated standard curve construction and sample preparation, can be performed in a single day of unattended operation. The method is linear (r greater than .99) in the range of 0 to 0.2 EU/ml. The precision of the method determined by assaying a lot of calcium gluconate for four days was 6%, 10%, and 10% for within an assay block, between assay blocks, and between assay days, respectively. Recovery of endotoxin when spiked into products ranged from 81% to 110% and was within the statistical variation (2 sigma limit) of the method. The endotoxin levels detected in a biological raw material by the chromogenic substrate assay method correlated well with that of the gel-clot LAL assay method. The endotoxin content of the majority of the pharmaceutical products tested was well below the sensitivity of both the chromogenic substrate and the gel clot LAL assay methods.

  14. 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

  15. Role of mononuclear phagocyte function in endotoxin-induced tumor necrosis

    NARCIS (Netherlands)

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

    1984-01-01

    The temporal susceptibility of tumors to induction of necrosis and regression by endotoxin was investigated further with a focus on the role of the putative mediator, tumor necrosis factor (TNF). Production of this factor was shown earlier to require prior activation of the mononuclear phagocytic sy

  16. 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

    -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......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...... low-grade inflammatory response in vivo. Two hundred healthy young male volunteers were genotyped, and cytokine levels were measured in response to a low-dose intravenous bolus of Escherichia coli endotoxin. No association was detected between SNPs (TLR-4299, TLR-4399, TNF-308, IL-18-137, IL-18...

  17. 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

  18. Activity of wild-type and hybrid Bacillus thuringiensis delta-endotoxins against Agrotis ipsilon

    NARCIS (Netherlands)

    Maagd, de R.A.; Weemen-Hendriks, M.; Molthoff, J.W.; Naimov, S.

    2003-01-01

    Twelve Cry1 and two Cry9 ?-endotoxins fromBacillus thuringiensis were tested for their activity against black cutworm (Agrotis ipsilon).A. ipsilon was not susceptible to many toxins, but three toxins had significant activity. Cry9Ca was the most toxic, followed by Cry1Aa and Cry1Fb. Hybrids between

  19. Combining charcoal and elemental black carbon analysis in sedimentary archives: Implications for past fire regimes, the pyrogenic carbon cycle, and the human-climate interactions

    Science.gov (United States)

    Thevenon, Florian; Williamson, David; Bard, Edouard; Anselmetti, Flavio S.; Beaufort, Luc; Cachier, Hélène

    2010-07-01

    This paper addresses the quantification of combustion-derived products in oceanic and continental sediments by optical and chemical approaches, and the interest of combining such methods for reconstructing past biomass burning activity and the pyrogenic carbon cycle. In such context, the dark particles > 0.2 µm 2 remaining after the partial digestion of organic matter are optically counted by automated image analysis and defined as charcoal, while the elemental carbon remaining after thermal and chemical oxidative treatments is quantified as black carbon (BC). The obtained pyrogenic carbon records from three sediment core-based case studies, (i) the Late Pleistocene equatorial Pacific Ocean, (ii) the mid-Holocene European Lake Lucerne, and (iii) the Late Holocene African Lake Masoko, are interpreted as proxy records of regional transportation mechanisms and biomass burning activities. The results show that the burial of dark carbon-rich particles in the 360 kyr-long record from the west equatorial Pacific is controlled by the combination of sea-level changes and low-latitude atmospheric circulation patterns (summer monsoon dynamics). However, the three fold increases in charcoal and BC sediment influxes between 53-43 and 12-10 kyr BP suggest that major shifts in fire activity occur synchronously with human colonization in the Indo/Pacific region. The coarse charcoal distribution from a 7.2 kyr record from Lake Lucerne in Switzerland closely matches the regional timing of major technical, land-use, and socio-economic changes during the Neolithic (between ca. 5.7 and 5.2 kyr BP and 4.9-4.5 kyr BP), the Bronze and Iron Ages (at ca. 3.3 and 2.4 kyr BP, respectively), and the industrialization (after AD 1838), pointing to the key impact of human activities on the sources, transportation processes and reservoirs of refractory carbon during the Holocene. In the tropical Masoko maar lake in Tanzania, where charcoal and BC records are highly sensitive to the local climate

  20. Piroxicam reverses endotoxin-induced hypotension in rats: contribution of vasoactive eicosanoids and nitric oxide.

    Science.gov (United States)

    Buharalioglu, C Kemal; Korkmaz, Belma; Cuez, Tuba; Sahan-Firat, Seyhan; Sari, Ayse Nihal; Malik, Kafait U; Tunctan, Bahar

    2011-09-01

    Nitric oxide (NO) produced by inducible NO synthase (iNOS) is responsible for endotoxin-induced vascular hyporeactivity and hypotension resulting in multiple organ failure. Endotoxic shock is also characterized by decreased expression of constitutive cyclooxygenase (COX-1), cytochrome P450 (CYP) 4A and endothelial NOS (eNOS). Our previous studies demonstrated that dual inhibition of iNOS and COX with a selective COX-2 inhibitor, NS-398, or a non-selective COX inhibitor, indomethacin, restores blood pressure presumably because of increased production of 20-hydroxyeicosatetraenoic acid (20-HETE) derived from arachidonic acid (AA) by CYP4A in endotoxaemic rats. The aim of this study was to investigate the effects of piroxicam, a preferential COX-1 inhibitor, on the endotoxin-induced changes in blood pressure, expression of COX-1, inducible COX (COX-2), CYP4A1, eNOS, iNOS and heat shock protein 90 (hsp90), and production of PGI(2), PGE(2), 20-HETE and NO. Injection of endotoxin (10 mg/kg, i.p.) to male Wistar rats caused a fall in blood pressure and an increase in heart rate associated with elevated renal 6-keto-PGF(1α) and PGE(2) levels as well as an increase in COX-2 protein expression. Endotoxin also caused an elevation in systemic and renal nitrite levels associated with increased renal iNOS protein expression. In contrast, systemic and renal 20-HETE levels and renal expression of eNOS, COX-1 and CYP4A1 were decreased in endotoxaemic rats. The effects of endotoxin, except for renal COX-1 and eNOS protein expression, were prevented by piroxicam (10 mg/kg, i.p.), given 1 hr after injection of endotoxin. Endotoxin did not change renal hsp90 protein expression. These data suggest that a decrease in the expression and activity of COX-2 and iNOS associated with an increase in CYP4A1 expression and 20-HETE synthesis contributes to the effect of piroxicam to prevent the hypotension during rat endotoxaemia.

  1. 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

  2. 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...

  3. Different sensitivity of rabbit heart and skeletal muscle to endotoxin-induced impairment of mitochondrial function.

    Science.gov (United States)

    Trumbeckaite, S; Opalka, J R; Neuhof, C; Zierz, S; Gellerich, F N

    2001-03-01

    The involvement of mitochondrial dysfunction in septic disturbances of tissues is controversial. The aim of this study was to investigate the effects of endotoxin-induced sepsis on the function of heart and skeletal muscle mitochondria. Rabbits were made septic by subcutaneous injection of endotoxin (lipopolysaccharide, LPS) from Escherichia coli at concentrations of 100 or 150 microg LPS.kg(-1) 24 h prior to the experiments. Mitochondrial respiration was measured in saponin-skinned muscle fibers and compared with photometrically detected activities of respiratory chain enzymes as well as with function of perfused hearts. In heart fibers a dosage of 100 microg LPS.kg(-1) caused a significant decrease of state 3-respiration for the substrates pyruvate (-38%), octanoyl-carnitine (-38%) and succinate (-30%) with correspondingly decreased respiratory control indexes (RCI). In addition, endotoxin caused a decreased temporal stability of the rate of state 3-respiration. At least in part these changes can be attributed to a reduced activity of complex I + III (-50%) of the respiratory chain. State 4-respiration rates were not significantly altered. The lowered state 3-respiration in heart mitochondria seems to contribute to the impairment of heart muscle function as detected by an increase of coronary vascular resistance (CVR) in endotoxin-treated hearts. Functional properties of mitochondria from M. Vastus lasteralis were not affected by 100 microg LPS.kg(-1) but a higher dosage of 150 microg LPS.kg(-1) caused decreased RCI for the substrates pyruvate (-29%) and octanoyl-carnitine (-32%). Also the activity of complex I + III was not significantly affected at lower dose of endotoxin but decreased (-42%) after treatment with 150 microg LPS.kg(-1). Results demonstrate the involvement of impaired mitochondria in the pathophysiology of septic organ failure and a tissue specificity of endotoxaemia.

  4. Structural and Dynamic Insights into a Glycine-Mediated Short Analogue of a Designed Peptide in Lipopolysaccharide Micelles: Correlation Between Compact Structure and Anti-Endotoxin Activity.

    Science.gov (United States)

    Datta, Aritreyee; Jaiswal, Nancy; Ilyas, Humaira; Debnath, Shibjyoti; Biswas, Kaushik; Kumar, Dinesh; Bhunia, Anirban

    2017-02-21

    In this study, we report an interaction study of a 13-residue analogue peptide VG13P (VARGWGRKCPLFG), derived from a designed VG16KRKP peptide (VARGWKRKCPLFGKGG), with a Lys6Gly mutation and removal of the last three residues Lys(14)-Gly(15)-Gly(16), in lipopolysaccharide (LPS), a major component of the outer membrane of Gram-negative bacteria and responsible for sepsis or septic shock. VG13P displays an enhanced anti-endotoxin property as evident from significant reduction in LPS-induced TNF-α gene expression levels in a monocytic cell line, while it retains almost unchanged antimicrobial activity as its parent VG16KRKP against Gram-negative bacterial as well as fungal pathogens. In addition, in vitro LPS binding properties of VG13P in comparison to its parent VG16KRKP also remained unhindered, suggesting that the flexible C-terminal end of VG16KRKP may not play a major role in its observed antibacterial and LPS binding properties. An NMR-resolved solution structure of VG13P in LPS reveals two consecutive β-turns: one at the N-terminus, followed by another at the central region, closely resembling a rocking chair. The crucial Lys6Gly mutation along with C-terminal truncation from VG16KRKP reorients the hydrophobic hub in VG13P in a unique way so as to fold the N-terminal end back on itself, forming a turn and allowing Val1 and Ala2 to interact with Leu11 and Phe12 to bring the hydrophobic residues closer together to form a more compact hub compared to its parent. The hub is further strengthened via CH-π interaction between Gly4 and Phe12. This accounts for its improved anti-endotoxin activity as well as to its uninterrupted antimicrobial activity.

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  11. 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.

  12. Resistance of essential fatty acid-deficient rats to endotoxin-induced increases in vascular permeability

    Energy Technology Data Exchange (ETDEWEB)

    Li, E.J.; Cook, J.A.; Spicer, K.M.; Wise, W.C.; Rokach, J.; Halushka, P.V. (Medical Univ. of South Carolina, Charleston (USA))

    1990-06-01

    Resistance to endotoxin in essential fatty acid-deficient (EFAD) rats is associated with reduced synthesis of certain arachidonic acid metabolites. It was hypothesized that EFAD rats would manifest decreased vascular permeability changes during endotoxemia as a consequence of reduced arachidonic acid metabolism. To test this hypothesis, changes in hematocrit (HCT) and mesenteric localization rate of technetium-labeled human serum albumin (99mTc-HSA) and red blood cells (99mTc-RBC) were assessed in EFAD and normal rats using gamma-camera imaging. Thirty minutes after Salmonella enteritidis endotoxin, EFAD rats exhibited less hemoconcentration as determined by % HCT than normal rats. Endotoxin caused a less severe change in permeability index in the splanchnic region in EFAD rats than in normal rats (1.2 +/- 0.6 x 10(-3)min-1 vs. 4.9 +/- 1.7 x 10(-3)min-1 respectively, P less than 0.05). In contrast to 99mTc-HSA, mesenteric localization of 99mTc-RBC was not changed by endotoxin in control or EFAD rats. Supplementation with ethyl-arachidonic acid did not enhance susceptibility of EFAD rats to endotoxin-induced splanchnic permeability to 99mTc-HSA. Leukotrienes have been implicated as mediators of increased vascular permeability in endotoxin shock. Since LTC3 formation has been reported to be increased in EFA deficiency, we hypothesized that LTC3 may be less potent than LTC4. Thus the effect of LTC3 on mean arterial pressure and permeability was compared to LTC4 in normal rats. LTC3-induced increases in peak mean arterial pressure were less than LTC4 at 10 micrograms/kg (39 +/- 5 mm Hg vs. 58 +/- 4 mm Hg respectively, P less than 0.05) and at 20 micrograms/kg (56 +/- 4 mm Hg vs. 75 +/- 2 mm Hg respectively, P less than 0.05). LY171883 (30 mg/kg), an LTD4/E4 receptor antagonist, attenuated the pressor effect of LTC4, LTD4, and LTC3.

  13. 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.

  14. Response of in vivo protein synthesis in T lymphocytes and leucocytes to an endotoxin challenge in healthy volunteers.

    Science.gov (United States)

    Januszkiewicz, A; Loré, K; Essén, P; Andersson, B; McNurlan, M A; Garlick, P J; Ringdén, O; Andersson, J; Wernerman, J

    2002-11-01

    In vivo determination of protein synthesis in immune cells reflects metabolic activity and immunological activation. An intravenous injection of endotoxin to healthy volunteers was used as a human sepsis model, and in vivo protein synthesis of T lymphocytes and leucocytes was measured. The results were related to plasma concentrations of selected cytokines, peripheral cell counts and subpopulations of immune cells. The subjects (n = 8 + 8) were randomized to an endotoxin (4 ng/kg) or a saline group. In vivo protein synthesis was determined twice: before and 1-2.5 h after the endotoxin/saline injection. Protein synthesis decreased in isolated T lymphocytes, but increased in leucocytes. Plasma levels of TNF-alpha, IL-8, IL-6, IL-1 ra and IL-10 were elevated, whereas IL-2 and IFN-gamma, produced predominantly by T lymphocytes, did not change in response to endotoxin. Neutrophils increased, whereas lymphocytes and monocytes decreased 2.5 h after the endotoxin injection. Flow cytometry revealed a drop in total CD3+ T lymphocytes and CD56+ natural killer cells, accompanied by an increase in CD15+ granulocytes. In summary, in vivo protein synthesis decreased in T lymphocytes, while the total leucocyte population showed a concomitant increase immediately after the endotoxin challenge. The changes in protein synthesis were accompanied by alterations in immune cell subpopulations and in plasma cytokine levels.

  15. Response of in vivo protein synthesis in T lymphocytes and leucocytes to an endotoxin challenge in healthy volunteers

    Science.gov (United States)

    Januszkiewicz, A; Loré, K; EsséN, P; Andersson, B; Mcnurlan, M A; Garlick, P J; RingdéN, O; Andersson, J; Wernerman, J

    2002-01-01

    In vivo determination of protein synthesis in immune cells reflects metabolic activity and immunological activation. An intravenous injection of endotoxin to healthy volunteers was used as a human sepsis model, and in vivo protein synthesis of T lymphocytes and leucocytes was measured. The results were related to plasma concentrations of selected cytokines, peripheral cell counts and subpopulations of immune cells. The subjects (n = 8 + 8) were randomized to an endotoxin (4 ng/kg) or a saline group. In vivo protein synthesis was determined twice: before and 1–2·5 h after the endotoxin/saline injection. Protein synthesis decreased in isolated T lymphocytes, but increased in leucocytes. Plasma levels of TNF-α, IL-8, IL-6, IL-1 ra and IL-10 were elevated, whereas IL-2 and IFN-γ, produced predominantly by T lymphocytes, did not change in response to endotoxin. Neutrophils increased, whereas lymphocytes and monocytes decreased 2·5 h after the endotoxin injection. Flow cytometry revealed a drop in total CD3+ T lymphocytes and CD56+ natural killer cells, accompanied by an increase in CD15+ granulocytes. In summary, in vivo protein synthesis decreased in T lymphocytes, while the total leucocyte population showed a concomitant increase immediately after the endotoxin challenge. The changes in protein synthesis were accompanied by alterations in immune cell subpopulations and in plasma cytokine levels. PMID:12390314

  16. Mathematical relationships between spore concentrations, delta-endotoxin levels, and entomotoxicity of Bacillus thuringiensis preparations produced in different fermentation media.

    Science.gov (United States)

    Vu, Khanh Dang; Tyagi, R D; Surampalli, R Y; Valéro, J R

    2012-11-01

    Mathematic relationships between spore concentrations, delta-endotoxin concentrations and entomotoxicity (Tx) of Bacillus thuringiensis var. kurstaki HD-1 (Btk HD-1) preparations produced in six different media were analysed. The relationship between delta-endotoxin and spore concentration and SpTx-spore (specific Tx per 1000 spore) and spore concentration produced in the different media (starch industry wastewater (SIW) with total solids (TS) concentration of 15g/L, SIW with TS of 30g/L, SIW supplemented with 0.2% (w/v) colloidal chitin, SIW supplemented with 1.25% (w/v) cornstarch and 0.2% (v/v) Tween 80, secondary sludge, and semi-synthetic medium) strictly followed the Power law. Tx and delta endotoxin concentration followed the exponential relation whereas a definite relation between Tx and spore concentration could not be established. Spore and delta-endotoxin produced at the early time (12h) during fermentation might be more toxic than those produced during latter period of fermentation irrespective of media used. Tx and delta-endotoxin concentration exhibited a semi-log linear relationship. Based on these findings, delta-endotoxin concentration can be determined rapidly to monitor the progress of the biopesticide production process.

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

    DEFF Research Database (Denmark)

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

    2002-01-01

    Endotoxins of intestinal origin are supposed to play an important role in the development of alcoholic hepatitis in man. To estimate the role of immunoglobulin response to gut-derived endotoxin in the development of alcohol-induced liver disease, serum levels of IgA and IgG against fecal endotoxin......, 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...... 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...

  18. Probiotic yogurt in the elderly with intestinal bacterial overgrowth: endotoxaemia and innate immune functions.

    Science.gov (United States)

    Schiffrin, Eduardo J; Parlesak, Alexandr; Bode, Christiane; Bode, J Christian; van't Hof, Martin A; Grathwohl, Dominik; Guigoz, Yves

    2009-04-01

    A study was conducted in healthy elderly living independently in senior housing to assess the impact of a probiotic yoghurt supplement on small intestinal bacterial overgrowth. Twenty-three participants with positive and thirteen participants with negative hydrogen breath test were studied before and after a period of 4 weeks of probiotic yoghurt administration. Intestinal permeability, plasma endotoxin levels, phagocytic activity of leucocytes, cytokine production by monocytes and free radical response of neutrophils were determined. Intestinal permeability was similar for the two groups and was unaffected by probiotic treatment. Both plasma endotoxin levels and the basal phagocytic activity of leucocytes decreased after yoghurt intake in the two groups. Exposure of monocytes and neutrophils ex vivo led to an increased cytokine response and free radical response, respectively. The normalisation of the various cytokine responses was more apparent in the group with positive breath test. In addition, the plasma levels of lipoplysaccharide binding protein and soluble CD14, lipoplysaccharide pattern recognition receptors and surrogate markers of lipoplysaccharide permeability were diminished by the end of the study. In conclusion, probiotic administration in the elderly normalises the response to endotoxin, and modulates activation markers in blood phagocytes, and therefore may help reduce low-grade chronic inflammation.

  19. Protective effects of imipramine in murine endotoxin-induced acute lung injury.

    Science.gov (United States)

    Yang, Jin; Qu, Jie-ming; Summah, Hanssa; Zhang, Jin; Zhu, Ying-gang; Jiang, Hong-ni

    2010-07-25

    The tricyclic antidepressant imipramine has recently emerged as a cytoprotective agent, exerting beneficial effects in inflammatory tissue injury. The present study aimed to investigate therapeutic effects of imipramine in murine model of endotoxin-induced acute lung injury. Mice were administrated intraperitoneally with LPS (lipopolysaccharide) from Escherichia coli or vehicle. Imipramine was administrated intraperitoneally 30 min before LPS challenge. Pretreatment of mice with imipramine reduced lethality. Impramine also significantly attenuated lung inflammation, lung edema, MPO (myeloperoxidase) activity, lung tissue pathological changes and nuclear factor-kappaB DNA binding activity. The results of this study suggest that imipramine can exert protective effects in endotoxin-induced acute lung injury by suppressing nuclear factor-kappaB-mediated expression of inflammatory genes. Thus, imipramine could be a potential novel therapeutic agent for the treatment for acute lung injury.

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

    Directory of Open Access Journals (Sweden)

    Tatyana A. Kuznetsova

    2014-01-01

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

  1. Bacterial vaginosis -- aftercare

    Science.gov (United States)

    ... this page: //medlineplus.gov/ency/patientinstructions/000687.htm Bacterial vaginosis - aftercare To use the sharing features on this ... to back after you use the bathroom. Preventing Bacterial Vaginosis You can help prevent bacterial vaginosis by: Not ...

  2. Pregnancy Complications: Bacterial Vaginosis

    Science.gov (United States)

    ... Complications & Loss > Pregnancy complications > Bacterial vaginosis and pregnancy Bacterial vaginosis and pregnancy E-mail to a friend Please ... this page It's been added to your dashboard . Bacterial vaginosis (also called BV or vaginitis) is an infection ...

  3. Bone marrow-derived mesenchymal stem cells protect rats from endotoxin-induced acute lung injury

    Institute of Scientific and Technical Information of China (English)

    LIANG Zhi-xin; SUN Ji-ping; WANG Ping; TIAN Qing; YANG Zhen; CHEN Liang-an

    2011-01-01

    Background Acute lung injury (ALI) is a serious and common condition for which there are currently no specific strategies for treatment.Recent studies have suggested that bone marrow-derived multipotent mesenchymal stem cells (MSCs) may have therapeutic applications in multiple clinical disorders.We explored the biological effects of MSCs during endotoxin-induced ALl and the mechanisms involved.Methods MSCs were isolated from male rat bone marrow and the ALl model was induced by intravenous endotoxin injection.Female rats were sacrificed at 6 hours,24 hours,4 days,1 week and 3 weeks post-injection of MSCs or saline and the lung tissue,bronchoalveolar lavage fluid,and serum were harvested for analysis.We further evaluated the survival of the rats and examined the effects of endotoxin-induced injury on the interaction between alveolar macrophages (AMs) and MSCs in ex vivo.Results There was a significant decrease in numbers of neutrophils in bronchoalveolar lavage fluid (P <0.05),and myeloperoxidase activity in the lung (P<0.01),and of TNF-α and IL-1β in serum (P <0.05) in the MSC treated rats at 4 days.Furthermore,MSC treated rats exhibited improved survival,lower lung injury score,higher concentration of IL-10 in the serum and a reduced hydroxyproline content,but these differences were not statistically significant.Moreover,co-cultures of MSCs and AMs had significantly reduced levels of TNF-α,IL-1β and macrophage inflammatory protein (MIP)-1α and significantly increased levels of IL-10 (P<0.05) in the culture supernatants.Conclusions Treatment with intravenous injection of bone marrow-derived MSCs have beneficial effects on endotoxin-induced ALl in rats.The beneficial effect might be achieved through the engraftment of differentiated MSCs in the lungs and appears derive more from their capacity to secrete soluble factors that modulate immune responses.

  4. 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.

  5. 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

  6. 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....

  7. 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.

  8. 天然内毒素结合蛋白衍生肽拮抗内毒素的研究进展%Antagonizing Effect of Peptides Derived from Natural Endotoxin Binding Protein against Endotoxin

    Institute of Scientific and Technical Information of China (English)

    段光杰

    2012-01-01

    Endotoxin,a major causative agent of sepsis,activates host immune cells by triggering of toll like receptor 4 transmembrane signaling,which secrete pro-inflammatory mediators and cytokines,resulting in tissues injury, multiple organ failure and even death. Thereby, antagonizing the activity of endotoxin is considered as the key point on intervention against sepsis. It is one of the important ways to attenuate the endotoxin-induced inflammatory responses using endotoxin neutralizing proteins/peptides,which could be directly synthesized or extracted from natural endotoxin binding protein. Here is to make a review on antagonizing effect of peptides derived from natural endotoxin binding protein against endotoxin.%内毒素是脓毒症最主要的致病因子,其可通过激活Toll样受体4跨膜信号通路,活化宿主免疫细胞,促进细胞因子和炎性介质的过度释放,导致组织损伤、多器官衰竭甚至死亡,因此,拮抗内毒素活性是提高脓毒症治疗效果的关键.从天然存在的内毒素结合蛋白中提取或者人工合成具有拮抗内毒素活性的蛋白/多肽类制剂是内毒素拮抗研究的主要方向之一.现对天然内毒素结合蛋白衍生肽拮抗内毒素的研究进展予以综述.

  9. 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.

  10. 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.

  11. The Role of Uncoupling Protein 2 During Myocardial Dysfunction in a Canine Model of Endotoxin Shock.

    Science.gov (United States)

    Wang, Xiaoting; Liu, Dawei; Chai, Wenzhao; Long, Yun; Su, Longxiang; Yang, Rongli

    2015-03-01

    To explore the role of uncoupling protein 2 (UCP2) during myocardial dysfunction in a canine model of endotoxin shock, 26 mongrel canines were randomly divided into the following four groups: A (control group; n = 6), B2 (shock after 2 h; n = 7), B4 (shock after 4 h; n = 7), and B6 (shock after 6 h; n = 6). Escherichia coli endotoxin was injected into the canines via the central vein, and hemodynamics were monitored. Energy metabolism, UCP2 mRNA and protein expression, and UCP2 localization were analyzed, and the correlation between energy metabolism changes, and UCP2 expression was determined. After the canine endotoxin shock model was successfully established, the expression of UCP2 mRNA and protein was found to increase, with later time points showing significant increases (P shock (P shock, and UCP2 may play an important role in this process. The negative correlation between UCP2 expression and energy metabolism requires further study, as the results might contribute to the treatment of sepsis with heart failure.

  12. 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.

  13. Mitochondrial DAMPs induce endotoxin tolerance in human monocytes: an observation in patients with myocardial infarction.

    Directory of Open Access Journals (Sweden)

    Irene Fernández-Ruiz

    Full Text Available Monocyte exposure to mitochondrial Danger Associated Molecular Patterns (DAMPs, including mitochondrial DNA (mtDNA, induces a transient state in which these cells are refractory to further endotoxin stimulation. In this context, IRAK-M up-regulation and impaired p65 activity were observed. This phenomenon, termed endotoxin tolerance (ET, is characterized by decreased production of cytokines in response to the pro-inflammatory stimulus. We also show that monocytes isolated from patients with myocardial infarction (MI exhibited high levels of circulating mtDNA, which correlated with ET status. Moreover, a significant incidence of infection was observed in those patients with a strong tolerant phenotype. The present data extend our current understanding of the implications of endotoxin tolerance. Furthermore, our data suggest that the levels of mitochondrial antigens in plasma, such as plasma mtDNA, should be useful as a marker of increased risk of susceptibility to nosocomial infections in MI and in other pathologies involving tissue damage.

  14. 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.

  15. Probiotic yogurt in the elderly with intestinal bacterial overgrowth: endotoxaemia and innate immune functions

    DEFF Research Database (Denmark)

    Schiffrin, E.J.; Parlesak, Alexandr; Bode, C.

    2009-01-01

    A study was conducted in healthy elderly living independently in senior housing to assess the impact of a probiotic yoghurt supplement on small intestinal bacterial overgrowth. Twenty-three participants with positive and thirteen participants with negative hydrogen breath test were studied before...... and was unaffected by probiotic treatment. Both plasma endotoxin levels and the basal phagocytic activity of leucocytes decreased after yoghurt intake in the two groups. Exposure of monocytes and neutrophils ex vivo led to an increased cytokine response and free radical response, respectively. The normalisation...

  16. Application of Fishbone Diagram in Risk Control of Pyrogen Reaction%鱼骨图在致热源反应风险控制中的应用

    Institute of Scientific and Technical Information of China (English)

    蒋忠伟; 吴凤莲; 王秦; 许平娟

    2014-01-01

    This paper mainly expounds the application of fishbone diagram analysis method in the prevention of complications caused by dialysis equipment applications, and puts forward to the potential risks, which could cause pyrogen reaction to the patients, such as risks in the maintenance of water treatment system, disinfection of the reverse osmosis membrane, disinfection of the ward pipeline, the use of dialysate, disinfection of the dialysis machine. Corresponding prevention measures are also introduced in this paper.%本文主要阐述了鱼骨图分析法在预防透析设备所致并发症的应用。以致热源反应为例,提出了水处理系统预处理的维护,反渗膜的消毒,病房管道的消毒,透析液的使用,透析机的消毒,透析耗材的使用等环节中可能引发患者致热源反应存在的风险,并陈述了预防的措施。

  17. Endotoxin in fine (PM 2.5) and coarse (PM 2.5-10) particle mass of ambient aerosols. A temporo-spatial analysis

    Science.gov (United States)

    Heinrich, Joachim; Pitz, Mike; Bischof, Wolfgang; Krug, Norbert; Borm, Paul J. A.

    Objectives: We collected fine (PM 2.5) and coarse (PM 2.5-10) particulate matter fractions in two areas ˜80 km apart and measured soluble endotoxin concentrations in both particle fractions. Here we report on temporo-spatial variation of endotoxin content in the collected particles. Methods: Dichotomous Anderson samplers were used to collect 21 weekly samples of PM 2.5 and PM 2.5-10 in both towns from January to June 2002. Each Teflon filter was water extracted and endotoxin was measured by a chromogenic Limulus Amoebocyte Lysate method. Endotoxin concentrations were expressed per mg of fine or mg of coarse mass and per sampled air volume (m 3). Results: For both cities, the mean endotoxin content in PM 2.5 was 1.2 EU mg -1; however the endotoxin content in the coarse fraction was ˜10 times higher compared to the fine mass fractions. Although endotoxin content is highly variable over time, a good correlation was observed between the two town sites for both fine ( r=0.85) and coarse PM ( r=0.88). The fluctuations of weekly endotoxin means were high in both areas suggesting a strong temporal dependence on particle source and composition. The endotoxin content in particles collected during May and June were two to four times higher than concentrations measured during the winter and early spring weeks. Conclusions: Ambient airborne endotoxin concentrations were detected in coarse and fine particle fraction, but 10-fold higher in the coarse PM. The strong seasonality and the week to week fluctuation of endotoxin content in PM indicate different biologic PM properties which might affect results of time series studies on short-term effects as well as in vitro studies and human exposure studies.

  18. 肉毒素和肿瘤坏死因子对大白鼠肝功能的影响%The effect of endotoxin and tumor necrosis factor on hepatic function of rats

    Institute of Scientific and Technical Information of China (English)

    曹显科; 张蒙

    2002-01-01

    Objective To investigate the effect of endotoxin and tumor necrosis factor (TNF) on hepatic function of rats. Method Prepare hepatic cell suspension by filling male SD rats' livers with collagenaseⅠ ,culture in DMEM medium, and add endotoxin and TNF in it. After 24 hours detect GPT, GOT in supernatant fluid. Result The activity of GPT and GOT in supernatant fluid increased after adding endotoxin and TNF . Conclusion The system of hepatic cell membrane could be damaged severely after acted by endotoxin and TNF.

  19. 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

  20. 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

  1. 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

  2. Plasma endothelin-1 and tumor necrosis factor-alpha concentrations in pregnant and cyclic rats after low-dose endotoxin infusion

    NARCIS (Netherlands)

    Faas, MM; Bakker, WW; Valkhof, N; Baller, JFW; Schuiling, GA

    1997-01-01

    Plasma endothelin-1 and tumor necrosis factor-alpha were determined in pregnant and cyclic rats after infusion of either endotoxin (1.0 mu g/kg of body weight) or saline solution. After endotoxin, but not after saline solution, administration there was a transient endothelin-1 response in pregnant r

  3. 40 CFR 180.1154 - CryIA(c) and CryIC derived delta-endotoxins of Bacillus thuringiensis var. kurstaki encapsulated...

    Science.gov (United States)

    2010-07-01

    ...-endotoxins of Bacillus thuringiensis var. kurstaki encapsulated in killed Pseudomonas fluorescens, and the... Bacillus thuringiensis var. kurstaki encapsulated in killed Pseudomonas fluorescens, and the expression plasmid and cloning vector genetic constructs. CryIA(c) and CryIC derived delta-endotoxins of...

  4. Seasonal Changes in Endotoxin Exposure and Its Relationship to Exhaled Nitric Oxide and Exhaled Breath Condensate pH Levels in Atopic and Healthy Children.

    Directory of Open Access Journals (Sweden)

    Gwo-Hwa Wan

    Full Text Available Endotoxin, a component of the cell walls of gram-negative bacteria, is a contaminant in organic dusts (house dust and aerosols. In humans, small amounts of endotoxin may cause a local inflammatory response. Exhaled nitric oxide (eNO levels, an inflammation indicator, are associated with the pH values of exhaled breath condensate (EBC. This study evaluated seasonal changes on indoor endotoxin concentrations in homes and the relationships between endotoxin exposure and eNO/EBC pH levels for healthy children and children with allergy-related respiratory diseases. In total, 34 children with allergy-related respiratory diseases and 24 healthy children were enrolled. Indoor air quality measurements and dust sample analysis for endotoxin were conducted once each season inside 58 surveyed homes. The eNO, EBC pH levels, and pulmonary function of the children were also determined. The highest endotoxin concentrations were on kitchen floors of homes of children with allergy-related respiratory diseases and healthy children, and on bedroom floors of homes of asthmatic children and healthy children. Seasonal changes existed in endotoxin concentrations in dust samples from homes of children with allergic rhinitis, with or without asthma, and in EBC pH values among healthy children and those with allergy-related respiratory diseases. Strong relationships existed between endotoxin exposure and EBC pH values in children with allergic rhinitis.

  5. The combination of PRRS virus and bacterial endotoxin as a model for multifactorial respiratory disease in pigs.

    Science.gov (United States)

    Van Gucht, Steven; Labarque, Geoffrey; Van Reeth, Kristien

    2004-12-08

    This paper reviews in vivo studies on the interaction between porcine reproductive and respiratory syndrome virus (PRRSV) and LPS performed in the authors' laboratory. The main aim was to develop a reproducible model to study the pathogenesis of PRRSV-induced multifactorial respiratory disease. The central hypothesis was that respiratory disease results from an overproduction of proinflammatory cytokines in the lungs. In a first series of studies, PRRSV was shown to be a poor inducer of TNF-alpha and IFN-alpha in the lungs, whereas IL-1 and the anti-inflammatory cytokine IL-10 were produced consistently during infection. We then set up a dual inoculation model in which pigs were inoculated intratracheally with PRRSV and 3-14 days later with LPS. PRRSV-infected pigs developed acute respiratory signs for 12-24h upon intratracheal LPS inoculation, in contrast to pigs inoculated with PRRSV or LPS only. Moreover, peak TNF-alpha, IL-1 and IL-6 titers were 10-100 times higher in PRRSV-LPS inoculated pigs than in the singly inoculated pigs and the cytokine overproduction was associated with disease. To further prove the role of proinflammatory cytokines, we studied the effect of pentoxifylline, a known inhibitor of TNF-alpha and IL-1, on PRRSV-LPS induced cytokine production and disease. The clinical effects of two non-steroidal anti-inflammatory drugs (NSAIDs), meloxicam and flunixin meglumine, were also examined. Pentoxifylline, but not the NSAIDs, significantly reduced fever and respiratory signs from 2 to 6h after LPS. The levels of TNF-alpha and IL-1 in the lungs of pentoxifylline-treated pigs were moderately reduced, but were still 26 and 3.5-fold higher than in pigs inoculated with PRRSV or LPS only. This indicates that pathways other than inhibition of cytokine production contributed to the clinical improvement. Finally, we studied a mechanism by which PRRSV may sensitize the lungs for LPS. We hypothesized that PRRSV would increase the amount of LPS receptor complex in the lungs leading to LPS sensitisation. Both CD14 and LPS-binding protein, two components of this complex, increased significantly during infection and the amount of CD14 in particular was correlated with LPS sensitisation. The increase of CD14 was mainly due to infiltration of strongly CD14-positive monocytes in the lungs. The PRRSV-LPS combination proved to be a simple and reproducible experimental model for multifactorial respiratory disease in pigs. To what extent the interaction between PRRSV and LPS contributes to the development of complex respiratory disease is still a matter of debate.

  6. CXCL8((3-73))K11R/G31P antagonizes the neutrophil chemoattractants present in pasteurellosis and mastitis lesions and abrogates neutrophil influx into intradermal endotoxin challenge sites in vivo.

    Science.gov (United States)

    Li, Fang; Zhang, Xiaobei; Mizzi, Chris; Gordon, John R

    2002-11-01

    The ELR(+) CXC chemokines are critical for protective neutrophil responses to most bacterial infections, but nevertheless can contribute importantly to the pathogenic effects of many inflammatory responses. We recently engineered a series of high affinity CXCL8/IL-8 antagonists, one of which, CXCL8((3-73))K11R/G31P, binds very strongly to neutrophils via the CXCR1 and CXCR2. Herein we show in competitive 125I-ligand binding assays that bovine CXCL8((3-73))K11R/G31P has an affinity for neutrophils that is 2-3 orders of magnitude higher than that of CXCL8/IL-8. Furthermore, when used at approximately 0.5 nM, CXCL8((3-73))K11R/G31P inhibited by 50% the chemotactic responses of neutrophils to 129 nM CXCL8/IL-8, but it also blocked chemotactic responses to the alternate ELR-CXC chemokines CXCL1/GRO alpha and CXCL5/ENA-78. Furthermore, CXCL8((3-73))K11R/G31P could inhibit by 93-97% the spectrum of neutrophil chemotactic activities present within wash fluids from clinical bacterial pneumonia or experimental endotoxin-induced mastitis lesions. Finally, intramuscular or subcutaneous application of CXCL8((3-73))K11R/G31P (75 micro g/kg) reduced by up to 97% neutrophil infiltration into intradermal endotoxin challenge sites in cattle, and prevented their circulating neutrophils from responding to CXCL8/IL-8 or ENA-78 in vitro. This data thus encourages further investigation of the potential impact of this novel antagonist on ELR-CXC chemokine-driven inflammatory disorders.

  7. Strong associations among rumen endotoxin and acute phase proteins with plasma minerals in lactating cows fed graded amounts of concentrate.

    Science.gov (United States)

    Zebeli, Q; Dunn, S M; Ametaj, B N

    2010-04-01

    The objective of this investigation was to determine associations among rumen endotoxin, plasma serum amyloid A (SAA), and C-reactive protein (CRP) with plasma Ca, Fe, Zn, and Cu in lactating cows challenged with graded amounts of rolled barley grain in the diet (i.e., 0, 15, 30, and 45% of DMI). Correlative relationships among variables were determined by linear and nonlinear regression procedures adjusted for the effects of day, animal, and experimental period. Increasing the amount of grain in the diet was successful in inducing an acute phase response, as assessed by augmentation of rumen endotoxin and plasma CRP and SAA (P minerals. Results suggest that new feeding strategies should be developed to curb the release of endotoxin in the rumen fluid to prevent perturbing minerals in the plasma.

  8. INFLUENCE OF NEOMYCIN AND INGESTED ENDOTOXIN IN THE PATHOGENESIS OF CHOLINE DEFICIENCY CIRRHOSIS IN THE ADULT RAT

    Science.gov (United States)

    Broitman, Selwyn A.; Gottlieb, Leonard S.; Zamcheck, Norman

    1964-01-01

    Two groups of adult rats fed a choline-deficient diet supplemented with neomycin in their drinking water for 250 or 350 days were protected against the development of liver fibrosis and cirrhosis. At the termination of the study these animals weighed more than others not receiving neomycin. This difference in weight did not appear to be caused by a growth-promoting effect of neomycin but rather reflected the increased severity of liver disease and a resultant weight loss in animals not receiving neomycin. Protection by neomycin was cancelled when Salmonella typhosa endotoxin was added to the drinking water. It was concluded that the protective effect of neomycin was mediated by an alteration in the intestinal microflora resulting in a reduction in the numbers of organisms contributing to intraluminal endotoxin. In the presence of choline deficiency, absorption of intraluminal endotoxin may contribute to the development of fibrosis and cirrhosis. PMID:14151103

  9. 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.

  10. 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.

  11. Cytotoxin and Pyrogenic Toxin Superantigen Gene Profiles of Staphylococcus aureus Associated with Subclinical Mastitis in Dairy Cows and Relationships with Macrorestriction Genomic Profiles

    Science.gov (United States)

    Fueyo, J. M.; Mendoza, M. C.; Rodicio, M. R.; Muñiz, J.; Alvarez, M. A.; Martín, M. C.

    2005-01-01

    A set of 84 Staphylococcus aureus isolates collected from the milk of cows with subclinical mastitis in Asturias (a cattle region of Spain) and six control strains were tested for sequences of genes encoding hemolysins (hla, hlb, hld, hlg, and hlg-2), leukotoxins (lukPV, lukM, and lukED), toxic shock syndrome toxin (tst), and enterotoxins (sea to see, seg to ser, and seu) by conventional and multiplex PCR. It was found that 84, 83, 11, and 39 isolates carried some type of hl, luk, tst, or se gene, respectively, which were arranged in 14 exotoxin genotypes. All of the isolates were negative for lukPV, hlg, sea, sed, see, sej, sek, sep, seq, and ser. Two gene groupings could be related with pathogenicity islands—[lukED, seg, sei, sem, sen, seo ± seu] with Saβ-1 and [tst, sec, sel] with SaPIbov, present in 45 and 13.1% of the isolates, respectively—while 11.9% of them carried both islands. Only one contained seb (together with υSaβ-1), and another contained seh (together with lukED). The isolates were also analyzed by pulsed-field gel electrophoresis performed with SmaI. Thirty-nine SmaI profiles (similarity coefficient [S] = 0.94 to 0.21) were differentiated; 12, 1, and 10 of these, respectively, were generated by isolates presumptively carrying Saβ-1, SaPIbov, or both. Five SmaI profiles (S ≥ 0.8) formed a cluster, which contained 20 and 10 isolates carrying one (υSaβ-1) or both islands. These data show the high frequency of genes encoding cytotoxins and pyrogenic toxin superantigens, their relationship with pathogenicity islands, and their distribution among a diversity of genetic types of S. aureus related to subclinical mastitis. PMID:15750096

  12. 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).

  13. 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.

  14. 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.

  15. Immunological enhancement action of endotoxin-free tilapia heat shock protein 70 against Streptococcus iniae.

    Science.gov (United States)

    Chen, Ming; Wang, Rui; Li, Liping; Liang, Wanwen; Wang, Qiuhua; Huang, Ting; Li, Chao; Li, Jian; Gan, Xi; Lei, Aiying; Huang, Weiyi; Luo, Honglin

    2014-07-01

    The immunological effects of heat shock proteins (HSPs) had been found in humans and mice, but scarce data of endotoxin-free Hsp70 were reported in tilapia. In the current study, we reported that tHsp70 alone and antigen-tHsp70 compound increased the proliferations of lymphocytes and macrophages, significantly increased the NO release and phagocytotic ability of macrophages (ptilapia lymphocytes and macrophages post S. iniae exposure and its up-regulation effects on vaccine-induced protection. Our research highlights the immunological enhancement action of Hsp70 in teleost immunity.

  16. 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.

  17. 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

    2016-06-14

    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.

  18. 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

  19. 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)

  20. Disorders of ventricular contractility and electrogenesis in the early stage of endotoxin shocked rabbits.

    Science.gov (United States)

    Lolov, R; Velkov, Z

    1994-01-01

    This is a report on ventricular contractility and electrogenesis disorders in rabbits, following intravenous injection of E. coli endotoxin at a dose of 2 mg.kg-1. At the 30th min, the right ventricular contractility indices (dP/dtmax)/P and [(dP/dt)/P]max had lower values, whereas end diastolic pressure (EDP), right ventricular systolic pressure (RVSP) and P(dP/dtmax) showed higher values compared to the initial ones. Most of the left ventricular contractility indices tested showed significantly lower values at the 30th and 60th min of the registration. In the scalar orthogonal ECG leads, at the 5th min an increased Qz amplitude, and at the 60th min an increased Rz amplitude, a decreased Ry amplitude, and QRS complex widening and bradicardia, were registered. In the spatial magnitude curve an increased amplitude of the main vectors of ventricular depolarization was documented. The changes in electrogenesis are interpreted first and foremost by the presence of hemodynamic disorders. The inference is reached that both left and right ventricular dysfunction have been already formed during the initial stage of endotoxin shock.

  1. Exosome-delivered microRNAs modulate the inflammatory response to endotoxin.

    Science.gov (United States)

    Alexander, Margaret; Hu, Ruozhen; Runtsch, Marah C; Kagele, Dominique A; Mosbruger, Timothy L; Tolmachova, Tanya; Seabra, Miguel C; Round, June L; Ward, Diane M; O'Connell, Ryan M

    2015-06-18

    MicroRNAs regulate gene expression posttranscriptionally and function within the cells in which they are transcribed. However, recent evidence suggests that microRNAs can be transferred between cells and mediate target gene repression. We find that endogenous miR-155 and miR-146a, two critical microRNAs that regulate inflammation, are released from dendritic cells within exosomes and are subsequently taken up by recipient dendritic cells. Following uptake, exogenous microRNAs mediate target gene repression and can reprogramme the cellular response to endotoxin, where exosome-delivered miR-155 enhances while miR-146a reduces inflammatory gene expression. We also find that miR-155 and miR-146a are present in exosomes and pass between immune cells in vivo, as well as demonstrate that exosomal miR-146a inhibits while miR-155 promotes endotoxin-induced inflammation in mice. Together, our findings provide strong evidence that endogenous microRNAs undergo a functional transfer between immune cells and constitute a mechanism of regulating the inflammatory response.

  2. 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.

  3. Dexamethasone and indomethacin modify endotoxin-induced respiratory failure in pigs.

    Science.gov (United States)

    Olson, N C; Brown, T T; Anderson, D L

    1985-01-01

    We studied the porcine pulmonary response to endotoxemia before and after administration of nonsteroidal antiinflammatory drugs (NSAID, i.e., indomethacin or flunixin meglumine) or dexamethasone (DEX). Escherichia coli endotoxin was infused intravenously into anesthetized 10- to 12-wk old pigs for 4.5 h. In endotoxemic pigs, the phase 1 (i.e., 0-2 h) increases in pulmonary arterial pressure, pulmonary vascular resistance (PVR), and alveolar-arterial O2 gradient and the decreases in cardiac index (CI) and lung dynamic compliance (Cdyn) were blocked by NSAID. Thus phase 1 changes were cyclooxygenase dependent. Furthermore, these effects were blocked or greatly attenuated by DEX. During phase 2 of endotoxemia (i.e., 2-4.5 h), the increased PVR and decreased CI and Cdyn were not blocked by NSAID but were attenuated by DEX, suggesting the presence of cyclooxygenase-independent metabolites. Both NSAID and DEX blocked the endotoxin-induced increases in lung water, bronchoalveolar lavage (BAL) neutrophil, and BAL albumin content. The fall in plasma proteins persisted in NSAID but not DEX-treated pigs. We conclude that endotoxemia in the pig causes severe acute respiratory failure largely mediated by cyclooxygenase and possibly lipoxygenase products of arachidonic acid metabolism.

  4. 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.

  5. The effect of endotoxin on the controllability of cardiac rhythm in rats.

    Science.gov (United States)

    Mazloom, Roham; Shirazi, Amir H; Hajizadeh, Sohrab; Dehpour, Ahmad R; Mani, Ali R

    2014-03-01

    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