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Sample records for endotoxin tolerance selective

  1. Dynamic and selective nucleosome repositioning during endotoxin tolerance.

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    El Gazzar, Mohamed; Liu, Tiefu; Yoza, Barbara K; McCall, Charles E

    2010-01-08

    Sepsis is encoded by a sequel of transcription activation and repression events that initiate, sustain, and resolve severe systemic inflammation. The repression/silencing phase occurs in blood leukocytes of animals and humans following the initiation of systemic inflammation due to developing endotoxin tolerance. We previously reported that NF-kappaB transcription factor RelB and histone H3 lysine methyltransferase G9a directly interact to induce facultative heterochromatin assembly and regulate epigenetic silencing during endotoxin tolerance, which is a major feature of sepsis. The general objective of this study was to assess whether dynamic temporal, structural, and positional changes of nucleosomes influence the sepsis phenotype. We used the THP-1 sepsis cell model to isolate mononucleosomes by rapid cell permeabilization and digestion of chromatin with micrococcal nuclease and then compared tumor necrosis factor alpha (TNFalpha) proximal promoter nucleosome alignment in endotoxin-responsive and -tolerant phenotypes. We found differential and dynamic repositioning of nucleosomes from permissive to repressive locations during the activation and silencing phases of transcription reprogramming and identified the following mechanisms that may participate in the process. 1) Two proximal nucleosomes repositioned to expose the primary NF-kappaB DNA binding site in endotoxin-responsive cells, and this "promoter opening" required the ATP-independent chaperone NAP1 to replace the core histone H2A with the H2A.Z variant. 2) During RelB-dependent endotoxin tolerance, the two nucleosomes repositioned and masked the primary NF-kappaB DNA binding site. 3) Small interfering RNA-mediated inhibition of RelB expression prevented repressive nucleosome repositioning and tolerance induction, but the "open" promoter required endotoxin-induced NF-kappaB p65 promoter binding to initiate transcription, supporting the known requirement of p65 posttranslational modifications for

  2. NAD+-dependent SIRT1 deacetylase participates in epigenetic reprogramming during endotoxin tolerance.

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    Liu, Tie Fu; Yoza, Barbara K; El Gazzar, Mohamed; Vachharajani, Vidula T; McCall, Charles E

    2011-03-18

    Gene-selective epigenetic reprogramming and shifts in cellular bioenergetics develop when Toll-like receptors (TLR) recognize and respond to systemic life-threatening infections. Using a human monocyte cell model of endotoxin tolerance and human leukocytes from acute systemic inflammation with sepsis, we report that energy sensor sirtuin 1 (SIRT1) coordinates the epigenetic and bioenergy shifts. After TLR4 signaling, SIRT1 rapidly accumulated at the promoters of TNF-α and IL-1β, but not IκBα; SIRT1 promoter binding was dependent on its co-factor, NAD(+). During this initial process, SIRT1 deacetylated RelA/p65 lysine 310 and nucleosomal histone H4 lysine 16 to promote termination of NFκB-dependent transcription. SIRT1 then remained promoter bound and recruited de novo induced RelB, which directed assembly of the mature transcription repressor complex that generates endotoxin tolerance. SIRT1 also promoted de novo expression of RelB. During sustained endotoxin tolerance, nicotinamide phosphoribosyltransferase (Nampt), the rate-limiting enzyme for endogenous production of NAD(+), and SIRT1 expression increased. The elevation of SIRT1 required protein stabilization and enhanced translation. To support the coordination of bioenergetics in human sepsis, we observed elevated NAD(+) levels concomitant with SIRT1 and RelB accumulation at the TNF-α promoter of endotoxin tolerant sepsis blood leukocytes. We conclude that TLR4 stimulation and human sepsis activate pathways that couple NAD(+) and its sensor SIRT1 with epigenetic reprogramming.

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

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    Ewelina Wiśnik

    2017-03-01

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

  4. 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.

  5. A selective sorbent for removing bacterial endotoxins from blood

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    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.

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

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    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.

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

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    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.

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

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    Ying Sun

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

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

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    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.

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

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    Loosbroock C

    2014-12-01

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

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

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

    2012-01-01

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

  12. RelB sustains IkappaBalpha expression during endotoxin tolerance.

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    Chen, Xiaoping; Yoza, Barbara K; El Gazzar, Mohamed; Hu, Jean Y Q; Cousart, Sue L; McCall, Charles E

    2009-01-01

    Transcription factors and chromatin structural modifiers induce clinically relevant epigenetic modifications of blood leukocytes during severe systemic inflammation (SSI) in humans and animals. These changes affect genes with distinct functions, as exemplified by the silencing of a set of acute proinflammatory genes and the sustained expression of a group of antimicrobial and anti-inflammatory genes. This paradigm is closely mimicked in the THP-1 human promonocyte cell model of lipopolysaccharide (LPS) endotoxin tolerance. We previously reported that LPS-induced de novo expression of RelB is required for generating tolerance to interleukin-1beta (IL-1beta) and tumor necrosis factor alpha (TNF-alpha) expression. RelB represses transcription by binding with heterochromatic protein 1 alpha (HP1alpha) to the proximal promoters of IL-1beta and TNF-alpha. In contrast, we report herein that RelB is required for sustained expression of anti-inflammatory IkappaBalpha in LPS-tolerant THP-1 cells. RelB transcription activation requires binding to the IkappaBalpha proximal promoter along with NF-kappaB p50 and is associated with an apparent dimer exchange with p65. We also observed that RelB induced during human SSI binds to the IkappaBalpha proximal promoter of circulating leukocytes. We conclude that RelB functions as a dual transcription regulator during LPS tolerance and human SSI by activating and repressing innate immunity genes.

  13. Protein Phosphatase 2A Catalytic Subunit α Plays a MyD88-Dependent, Central Role in the Gene-Specific Regulation of Endotoxin Tolerance

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    Ling Xie

    2013-03-01

    Full Text Available MyD88, the intracellular adaptor of most TLRs, mediates either proinflammatory or immunosuppressive signaling that contributes to chronic inflammation-associated diseases. Although gene-specific chromatin modifications regulate inflammation, the role of MyD88 signaling in establishing such epigenetic landscapes under different inflammatory states remains elusive. Using quantitative proteomics to enumerate the inflammation-phenotypic constituents of the MyD88 interactome, we found that in endotoxin-tolerant macrophages, protein phosphatase 2A catalytic subunit α (PP2Ac enhances its association with MyD88 and is constitutively activated. Knockdown of PP2Ac prevents suppression of proinflammatory genes and resistance to apoptosis. Through site-specific dephosphorylation, constitutively active PP2Ac disrupts the signal-promoting TLR4-MyD88 complex and broadly suppresses the activities of multiple proinflammatory/proapoptotic pathways as well, shifting proinflammatory MyD88 signaling to a prosurvival mode. Constitutively active PP2Ac translocated with MyD88 into the nuclei of tolerant macrophages establishes the immunosuppressive pattern of chromatin modifications and represses chromatin remodeling to selectively silence proinflammatory genes, coordinating the MyD88-dependent inflammation control at both signaling and epigenetic levels under endotoxin-tolerant conditions.

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

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

    2017-03-01

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

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

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

    2016-11-18

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

  16. [Experience of using endotoxin selective adsorption in patients with severe sepsis after open-heart surgery].

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    Iarustovskiĭ, M B; Abramian, M V; Krotenko, N P; Popov, D A; Pliushch, M G; Nazarova, E I; Gordeev, S L

    2014-01-01

    To evaluate the safety and effectiveness of selective lipopolysaccharide (LPS)-adsorption therapy using polymyxin B immobilised fibre cartridges in adult patients complicated with severe sepsis after cardiac surgery. 105 patients received extracorporeal LPS-adsorption procedures using Toraymyxin columns--PMX (Toray, Japan) in addition to the standard treatment according to the Surviving Sepsis Campaign guideline study group. For control group we selected 40 patients, comparable by PMX group in age, body weight, severity of illness, and the duration of cardiopulmonary bypass, received only standard therapy. All patients received significant doses of vasoactive drugs for hemodynamic support, mechanical ventilation and broad-spectrum antibiotics. Mean APACHE II and SOFA scores were comparable for both groups. Inclusion criteria were: clinical signs of severe sepsis, endotoxin activity assay (EAA) > or = 0.6, elevated blood plasma procalcitonin (PCT) > 2 ng ml(-1). The inclusion criteria were clinical signs of severe sepsis, endotoxin activity assay (EAA) > or = 0.6, and blood plasma procalcitonin (PCT) > 2 ng ml(-1). Extracorporeal treatment was administered within 24 h of a severe sepsis diagnosis. Each patient in PMX group received 2 LPS-adsorption procedures and each session of hemoperfusion lasted for 120 minutes. After the LPS-adsorption course, we noted any indices of haemodynamic improvements, including an increase in mean arterial pressure on 22% (p < 0.001), mean oxygenation index (on 24.5%, p < 0.001), normalisation of leukocytosis and a decrease in mean body temperature. After the procedures of LPS-adsorption we found the statistically significant decreasing of LPS concentrations according to LAL-test and EAA. In the control group, there were no significant changes in any of the studied parameters except body temperature. Moreover, the 28-day mortality was 42% in the study group and 65% in the control group (p = 0.032). The endotoxin adsorption procedures

  17. The NF-kappaB factor RelB and histone H3 lysine methyltransferase G9a directly interact to generate epigenetic silencing in endotoxin tolerance.

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    Chen, Xiaoping; El Gazzar, Mohamed; Yoza, Barbara K; McCall, Charles E

    2009-10-09

    The interplay of transcription factors, histone modifiers, and DNA modification can alter chromatin structure that epigenetically controls gene transcription. During severe systemic inflammatory (SSI), the generation of facultative heterochromatin from euchromatin reversibly silences transcription of a set of acute proinflammatory genes. This gene-specific silencing is a salient feature of the endotoxin tolerant phenotype that is found in blood leukocytes of SSI patients and in a human THP-1 cell model of SSI. We previously reported that de novo induction of the NF-kappaB transcription factor RelB by endotoxin activation is necessary and sufficient for silencing transcription of acute proinflammatory genes in the endotoxin tolerant SSI phenotype. Here, we examined how RelB silences gene expression and found that RelB induces facultative heterochromatin formation by directly interacting with the histone H3 lysine 9 methyltransferase G9a. We found that heterochromatin protein 1 (HP1) and G9a formed a complex at the interleukin-1beta promoter that is dependent on the Rel homology domain (RHD) of RelB. RelB knockdown disassociated the complex and reversed transcription silencing. We also observed that whereas RelB chromatin binding was independent of G9a, RelB transcriptional silencing required G9a accumulation at the silenced promoter. Binding between RelB and G9a was confirmed by glutathione S-transferase pulldown in vitro and coimmunoprecipitation in vivo. These data provide novel insight into how RelB is required to initiate silencing in the phenotype associated with severe systemic inflammation in humans, a disease with major morbidity and mortality.

  18. Mechanisms of interleukin-2-induced hydrothoraxy in mice: protective effect of endotoxin tolerance and dexamethasone and possible role of reactive oxygen intermediates.

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    Faggioni, R; Allavena, P; Cantoni, L; Carelli, M; Demitri, M T; Delgado, R; Gatti, S; Gnocchi, P; Isetta, A M; Paganin, C

    1994-04-01

    Interleukin (IL)-2 is known to induce vascular leak syndrome (VLS), which was suggested to be mediated by immune system-derived cytokines, including tumor necrosis factor (TNF). To characterize the role of TNF in IL-2 toxicity in C3H/HeN mice, we used two approaches to downregulate TNF production in vivo: treatment with dexamethasone (DEX) and induction of endotoxin (lipopolysaccharide) (LPS) tolerance by a 4-day pretreatment with LPS (35 micrograms/mouse/day). Mice were then treated with IL-2 for 5 days (1.8 x 10(5) IU/mouse, twice daily). Both DEX and LPS tolerance blocked development of hydrothorax in IL-2-treated mice and inhibited TNF induction. DEX and LPS tolerance also ameliorated IL-2 toxicity in terms of decrease in food intake and inhibited the increase of the acute-phase protein, serum amyloid A (SAA). The IL-2 activation of splenic natural killer (NK) cell activity was also diminished by DEX and, to a lesser extent, by LPS-tolerance. Treatment with IL-2 also caused induction of the superoxide-generating enzyme xanthine oxidase (XO) in tissues and serum and induced bacterial translocation in the mesenteric lymph nodes (MLN). These data suggest that multiple mechanisms, including NK cell activity, cytokines, and reactive oxygen intermediates, might be important in the vascular toxicity of IL-2.

  19. Selective HDAC6 inhibition prevents TNF-α-induced lung endothelial cell barrier disruption and endotoxin-induced pulmonary edema.

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    Yu, Jinyan; Ma, Zhongsen; Shetty, Sreerama; Ma, Mengshi; Fu, Jian

    2016-07-01

    Lung endothelial damage contributes to the pathogenesis of acute lung injury. New strategies against lung endothelial barrier dysfunction may provide therapeutic benefits against lung vascular injury. Cell-cell junctions and microtubule cytoskeleton are basic components in maintaining endothelial barrier integrity. HDAC6, a deacetylase primarily localized in the cytoplasm, has been reported to modulate nonnuclear protein function through deacetylation. Both α-tubulin and β-catenin are substrates for HDAC6. Here, we examined the effects of tubastatin A, a highly selective HDAC6 inhibitor, on TNF-α induced lung endothelial cell barrier disruption and endotoxin-induced pulmonary edema. Selective HDAC6 inhibition by tubastatin A blocked TNF-α-induced lung endothelial cell hyperpermeability, which was associated with increased α-tubulin acetylation and microtubule stability. Tubastatin A pretreatment inhibited TNF-α-induced endothelial cell contraction and actin stress fiber formation with reduced myosin light chain phosphorylation. Selective HDAC6 inhibition by tubastatin A also induced β-catenin acetylation in human lung endothelial cells, which was associated with increased membrane localization of β-catenin and stabilization of adherens junctions. HDAC6 knockdown by small interfering RNA also prevented TNF-α-induced barrier dysfunction and increased α-tubulin and β-catenin acetylation in endothelial cells. Furthermore, in a mouse model of endotoxemia, tubastatin A was able to prevent endotoxin-induced deacetylation of α-tubulin and β-catenin in lung tissues, which was associated with reduced pulmonary edema. Collectively, our data indicate that selective HDAC6 inhibition by tubastatin A is a potent approach against lung endothelial barrier dysfunction. Copyright © 2016 the American Physiological Society.

  20. The role of heat-tolerant endotoxin-retentive ultrafilters (UFs) for the remediation of reverse osmosis (RO) plants employed for surgical hand antisepsis using periodic thermal disinfection -A ten-year longitudinal experience study in the operating theater.

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    Uetera, Yushi; Yasuhara, Hiroshi; Kumada, Naohito; Misawa, Yoshiki; Terada, Yoshiyuki; Satou, Yoshio; Suhara, Tamaki; Kawamura, Kunio

    2018-03-15

    Recently, the use of filters has come into light for sanitizing water plants. This study investigated the role of heat-tolerant ultrafilters (UFs) for the remediation of reverse osmosis (RO) plants using periodic thermal disinfection. Two completely identical RO plants (RO plants A & B) were installed in 2006 for surgical hand antisepsis in the operating theater. RO water was stored in the 300-liter storage tank and re-circulated in the 190 meter-long loop delivering water to 12 faucets in each RO plant. Periodic thermal disinfection has come into practice since a UF module was retrofitted to the recirculation loop of each RO plant in 2010. Endotoxin was monitored closely before and after thermal disinfection. Before UF modules were retrofitted, endotoxin increased to a maximum of 0.301 EU/ml in the RO plant A and 1.446 EU/ml in the RO plant B after thermal disinfection, respectively. Since a UF module was retrofitted to each RO plant in 2010, endotoxin has been continuously below 0.025 EU/ml in the RO plant A and exceeded this level five times in the RO plant B. On one occasion, endotoxin increased in all samples collected simultaneously after solenoid valves were replaced in the recirculation loop near the air conditioner outlet. At this time, the inside of pipework was exposed to the ventilation airflow. After the valves were replaced again with the work place isolated using a curing sheet, endotoxin decreased. On the other occasions, endotoxin increased only in one sample and decreased after thermal disinfection. Annually replaced UF modules were examined twice for estimating the amounts of immobilized endotoxin. The estimated amounts decreased in 2013 by the order of 10-3 in comparison with those in 2011 in both RO plants. The present study suggested that UFs acted synergistically with periodic thermal disinfection for the remediation of RO plants. Copyright © 2018, Parenteral Drug Association.

  1. THE EFFECTS OF BACTERIAL ENDOTOXIN ON LIPIDE METABOLISM

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    LeQuire, V. S.; Hutcherson, J. D.; Hamilton, R. L.; Gray, M. E.

    1959-01-01

    Single intravenous injections of Shear's polysaccharide in varying dosages invariably produced an elevation in the levels of the total serum lipides 24 hours after injection of endotoxin. The total serum cholesterol and lipide phosphorus were also affected, although they did not change with smaller doses of endotoxin and were rarely elevated to the same degree as were the total serum lipides. The degree of elevation of the serum lipides was apparently related to the amount of endotoxin injected up to a certain point, beyond which there was no further increase. There were two types of response to endotoxin by the serum lipides, a moderate increase and an uncontrolled increase. Higher dosages of endotoxin and fasting apparently increased the incidence of the latter response. No direct correlation could be made between serum lipide responses and histologic evidence typical of the generalized Shwartzman reaction following this regimen of endotoxin injection. The Shwartzman reaction did occur with greater frequency and with lower dosages of endotoxin in fasted animals. Animals given repeated injections of endotoxin showed an initial increase in serum lipides followed by a progressive decrease to normal levels as tolerance to the febrile action of endotoxin appeared. The febrile tolerance as well as the unresponsiveness of the serum lipides to endotoxin was abolished by thorium dioxide (thorotrast) in these animals. In similar experiments a "breakthrough" of lipide unresponsiveness to endotoxin was obtained by increasing the amount of endotoxin injected. Some of the implications of these results for the metabolic alterations produced by bacterial endotoxins are discussed. PMID:13673140

  2. TLR ligands, but not modulators of histone modifiers, can induce the complex immune response pattern of endotoxin tolerance in mammary epithelial cells

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    Günther, Juliane; Petzl, Wolfram; Zerbe, Holm; Schuberth, Hans-Joachim

    2016-01-01

    Excessive stimulation of the TLR4 axis through LPS reduces the expression of some cytokine genes in immune cells, while stimulating the expression of immune defense genes during a subsequent bacterial infection. This endotoxin tolerance (ET) is mediated via epigenetic mechanisms. Priming the udder of cows with LPS was shown to induce ET in mammary epithelial cells (MEC), thereby protecting the udder against reinfection for some time. Seeking alternatives to LPS priming we tried to elicit ET by priming MEC with either lipopeptide (Pam2CSK4) via the TLR2/6 axis or inhibitors of histone-modifying enzymes. Pre-incubation of MEC with Pam2CSK4 enhanced baseline and induced expression of bactericidal (β-defensin; SLPI) and membrane protecting factors (SAA3, TGM3), while reducing the expression of cytokine- and chemokine-encoding genes (TNF, IL1β) after a subsequent pathogen challenge, the latter, however, not as efficiently as after LPS priming. Pre-treating MEC with various inhibitors of histone H3 modifiers (for demethylation, acetylation or deacetylation) all failed to induce any of the protective factors and only resulted in some dampening of cytokine gene expression after the re-challenge. Hence, triggering immune functions via the TLR axis, but not through those histone modifiers, induced the beneficial phenomenon of ET in MEC. PMID:27913794

  3. Selection and characterisation of high ethanol tolerant ...

    African Journals Online (AJOL)

    15% ethanol tolerance. High level ethanol tolerant Saccharomyces yeast, Orc 6, was investigated for its potential application in ethanologenic fermentations. Data presented in this study revealed that Orc 6 yeast isolate tolerated osmotic stress above 12% (w/v) sorbitol and 15% (w/v) sucrose equivalent of osmotic pressure ...

  4. Selection on resilience improves disease resistance and tolerance to infections

    NARCIS (Netherlands)

    Mulder, H.A.; Rashidi, H.

    2017-01-01

    Response to infection in animals has 2 main mechanisms: resistance (ability to control pathogen burden) and tolerance (ability to maintain performance given the pathogen burden). Selection on disease resistance and tolerance to infections seems a promising avenue to increase productivity of animals

  5. Analysis of interleukin-6 in endotoxin-induced uveitis

    NARCIS (Netherlands)

    Hoekzema, R.; Murray, P. I.; van Haren, M. A.; Helle, M.; Kijlstra, A.

    1991-01-01

    The mechanisms underlying the induction of intraocular inflammation in the rat model of endotoxin-induced uveitis (EIU) and the subsequent development of tolerance after repeated endotoxin injections are poorly understood. Interleukin-6 (IL-6) was measured in the aqueous humor and serum of Lewis

  6. Rapid screening for selection of heavy metal-tolerant plants

    OpenAIRE

    Fernando Angelo Piotto; Augusto Tulmann-Neto; Mônica Regina Franco; Luis Felipe Boaretto; Ricardo Antunes Azevedo

    2014-01-01

    One of the major difficulties in studies on the selection of heavy metal tolerant plants is the proper methodology that must ensure an efficient evaluation of a large number of plants, but reducing environmental contamination. For this reason, we propose a simple and effective method that can be applied to metals or other chemicals by means of a case study, in which we selected two cadmium-tolerant mutants of tomato (cv Micro-Tom) obtained by mutagenesis with methyl ethanesulfonat...

  7. Children’s residential exposure to selected allergens and microbial indicators: endotoxins and (1→3-β-D-glucans

    Directory of Open Access Journals (Sweden)

    Anna Kozajda

    2013-12-01

    Full Text Available Objectives: The study was aimed at assessment of exposure to endotoxins, (1→3-β-D-glucans and mite, cockroach, cat, dog allergens present in settled dust in premises of children as agents which may be significantly correlated with the occurrence of allergic symptoms and diseases in children. Materials and Methods: The study covered 50 homes of one- or two-year-old children in Poland. Samples of settled dust were taken from the floor and the child's bed. The levels of (1→3-β-D-glucans (floor, endotoxins (floor and allergens of mite, cat, dog and cockroach (floor and bed were analyzed. Results: Average geometric concentrations (geometric standard deviation of endotoxins, (1→3-β-D-glucans, Der p1, Fel d1, Can f1 and Bla g1 in children homes were on the floor 42 166.0 EU/g (3.2, 20 478.4 ng/g (2.38, 93.9 ng/g (6.58, 119.8 ng/g (13.0, 288.9 ng/g (3.4, 0.72 U/g (4.4 and in their beds (only allergens 597.8 ng/g (14.2, 54.1 ng/g (4.4, 158.6 ng/g (3.1 0.6 U/g (2.9, respectively. When the floor was covered with the carpet, higher concentrations of endotoxins, (1→3-β-D-glucans and allergens (each type were found in the settled dust (p < 0.05. The trend was opposite in case of allergens (except dog analyzed from bed dust and significantly higher concentrations were found in the rooms with smooth floor (p < 0.05. Conclusions: Among the analyzed factors only the type of floor significantly modified both the level of biological indicators and allergens. The results of this study could be the base for verifying a hypothesis that carpeting may have a protective role against high levels of cockroach, dog and cat allergens.

  8. Rapid screening for selection of heavy metal-tolerant plants

    Directory of Open Access Journals (Sweden)

    Fernando Angelo Piotto

    2014-03-01

    Full Text Available One of the major difficulties in studies on the selection of heavy metal tolerant plants is the proper methodology that must ensure an efficient evaluation of a large number of plants, but reducing environmental contamination. For this reason, we propose a simple and effective method that can be applied to metals or other chemicals by means of a case study, in which we selected two cadmium-tolerant mutants of tomato (cv Micro-Tom obtained by mutagenesis with methyl ethanesulfonate (MES. Aside from these two new mutants, we selected 21 others with possible alterations in the response to this metal. Finally, it was concluded that the proposed system is ideal for selection studies on plants tolerant to heavy metals and possibly to other elements, due to the ease of assembly of the structure, low installation cost, minimal waste generation and the possibility of using different species and heavy metals.

  9. Selection of D-Alanine-Tolerant Rice Cells

    OpenAIRE

    Hisashi, Manabe; Koji, Ohira; Aizu Junior College of Fukushima Prefecture; Department of Agricultural Chemistry, Faculty of Agriculture, Tohoku University

    1984-01-01

    By repeating subculture of rice cells (parent cells) in a D-alanine containing medium, we could select rice cells which grew well in the D-alanine medium. The D-alanine-tolerant cells absorbed a fairly small amount of D-alanine from the medium and did not accumulate much D-alanine in the cells. Aggregation of D-alanine-tolerant cells was greater than that of parent cells. D-Alanine metabolism of D-alanine.-tolerant cells did not increase in comparison with parent cells.

  10. Material Selection for Accident Tolerant Fuel Cladding

    International Nuclear Information System (INIS)

    Pint, Bruce A.; Terrani, Kurt A.; Yamamoto, Yukinori; Snead, Lance Lewis

    2015-01-01

    Alternative cladding materials to Zr-based alloys are being investigated for accident tolerance, which can be defined as > 100X improvement (compared to Zr-based alloys) in oxidation resistance to steam or steam-H 2 environments at ≥1473 K (1200°C) for short times. After reviewing a wide range of candidates, current steam oxidation testing is being conducted on Mo, MAX phases and FeCrAl alloys. Recently reported low mass losses for Mo in steam at 800°C could not be reproduced. Both FeCrAl and MAX phase Ti 2 AlC form a protective alumina scale in steam. However, commercial Ti 2 AlC that was not single phase, formed a much thicker oxide at 1200°C in steam and significant TiO 2 , and therefore Ti 2 AlC may be challenging to form as a cladding or a coating. Alloy development for FeCrAl is seeking to maintain its steam oxidation resistance to 1475°C, while reducing its Cr content to minimize susceptibility to irradiation-assisted α' formation. The composition effects and critical limits to retaining protective scale formation at > 1400°C are still being evaluated.

  11. Material Selection for Accident Tolerant Fuel Cladding

    Energy Technology Data Exchange (ETDEWEB)

    Pint, Bruce A. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Terrani, Kurt A. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Yamamoto, Yukinori [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Snead, Lance Lewis [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)

    2015-01-01

    Alternative cladding materials to Zr-based alloys are being investigated for accident tolerance, which can be defined as > 100X improvement (compared to Zr-based alloys) in oxidation resistance to steam or steam-H2 environments at ≥ 1200°C for short times. After reviewing a wide range of candidates, current steam oxidation testing is being conducted on Mo, MAX phases and FeCrAl alloys. Recently reported low mass losses for Mo in steam at 800°C could not be reproduced. Both FeCrAl and MAX phase Ti2AlC form a protective alumina scale in steam. However, commercial Ti2AlC that was not single phase, formed a much thicker oxide at 1200°C in steam and significant TiO2, and therefore Ti2AlC may be challenging to form as a cladding or a coating. Alloy development for FeCrAl is seeking to maintain its steam oxidation resistance to 1475°C, while reducing its Cr content to minimize susceptibility to irradiation-assisted α´ formation. The composition effects and critical limits to retaining protective scale formation at > 1400°C are still being evaluated.

  12. Marker assisted selection and crop management for salt tolerance: A ...

    African Journals Online (AJOL)

    Marker assisted selection and crop management for salt tolerance: A review. D Singh, A Kumar, A Kumar, P Chauhan, V Kumar, N Kumar, A Singh, N Mahajan, P Sirohi, S Chand, B Ramesh, J Singh, P Kumar, R Kumar, RB Yadav, RK Naresh ...

  13. Endotoxins in surgical instruments of hip arthroplasty

    Directory of Open Access Journals (Sweden)

    Vania Regina Goveia

    2016-06-01

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

  14. 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.

  15. Generation of American elm trees with tolerance to Dutch elm disease through controlled crosses and selection

    Science.gov (United States)

    James M. Slavicek; Kathleen S. Knight

    2012-01-01

    The goal of our research and development efforts is to generate new and/or improved selections of the American elm (Ulmus americana L.) with tolerance/resistance to Dutch elm disease (DED). The approaches we are taking for this effort include: 1) controlled breeding using known DED -tolerant selections, 2) controlled breeding using DED-tolerant...

  16. Combining ability of grain sorghum lines selected for aluminum tolerance

    Directory of Open Access Journals (Sweden)

    Cicero Beserra Menezes

    2014-03-01

    Full Text Available The purpose of this study was to estimate combining ability of 58 sorghum lines previously selected for Aluminum (Al tolerance. One hundred sixty-five hybrids were evaluated at three levels of Al saturation (0%, 20% and 50% at the same season. General Combining Ability (GCA effects were significant for female lines for all three traits. GCA effects for male lines were significant only for plant height. Specific Combining Ability (SCA effects were significant only for flowering time. The ratio GCA to SCA was greater than the unity, indicating the prevalence of additive effects for the control of Al tolerance. F7, F14, F17, F20, F21, F24, F29, F31, F41, F42, F48, F51, F54 and F55 lines contributed to increase yield, while F29, F48 and F51 also contributed to reduce flowering time. M2 was the best male line since it contributed to increase yield and plant height, and to reduce flowering time.

  17. The effects of bacterial endotoxin on lipide metabolism. I. The responses of the serum lipides of rabbits to single and repeated injections of Shear's polysaccharide.

    Science.gov (United States)

    LEQUIRE, V S; HUTCHERSON, J D; HAMILTON, R L; GRAY, M E

    1959-08-01

    Single intravenous injections of Shear's polysaccharide in varying dosages invariably produced an elevation in the levels of the total serum lipides 24 hours after injection of endotoxin. The total serum cholesterol and lipide phosphorus were also affected, although they did not change with smaller doses of endotoxin and were rarely elevated to the same degree as were the total serum lipides. The degree of elevation of the serum lipides was apparently related to the amount of endotoxin injected up to a certain point, beyond which there was no further increase. There were two types of response to endotoxin by the serum lipides, a moderate increase and an uncontrolled increase. Higher dosages of endotoxin and fasting apparently increased the incidence of the latter response. No direct correlation could be made between serum lipide responses and histologic evidence typical of the generalized Shwartzman reaction following this regimen of endotoxin injection. The Shwartzman reaction did occur with greater frequency and with lower dosages of endotoxin in fasted animals. Animals given repeated injections of endotoxin showed an initial increase in serum lipides followed by a progressive decrease to normal levels as tolerance to the febrile action of endotoxin appeared. The febrile tolerance as well as the unresponsiveness of the serum lipides to endotoxin was abolished by thorium dioxide (thorotrast) in these animals. In similar experiments a "breakthrough" of lipide unresponsiveness to endotoxin was obtained by increasing the amount of endotoxin injected. Some of the implications of these results for the metabolic alterations produced by bacterial endotoxins are discussed.

  18. Selection criteria for drought tolerance at the vegetative phase in ...

    African Journals Online (AJOL)

    Identifying drought tolerant maize (Zea mays L.) at the vegetative stage is a meaningful effort at reducing cost and time of screening large number of maize genotypes for drought tolerance. The primary objectives of this study were to assess the effectiveness of vegetative traits in discriminating between drought tolerant and ...

  19. Endotoxins in surgical instruments of hip arthroplasty

    OpenAIRE

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

    2016-01-01

    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...

  20. 40 CFR 180.1107 - Delta endotoxin of Bacillus thuringiensis variety kurstaki encapsulated into killed Pseudomonas...

    Science.gov (United States)

    2010-07-01

    ... thuringiensis variety kurstaki encapsulated into killed Pseudomonas fluorescens; exemption from the requirement... killed Pseudomonas fluorescens; exemption from the requirement of a tolerance. The delta endotoxin of Bacillus thuringiensis variety kurstaki encapsulated into killed Pseudomonas fluorescens is exempt from the...

  1. 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.

  2. Evaluation of drought tolerance indices for the selection of Iranian ...

    African Journals Online (AJOL)

    Ten barley genotypes were tested in a randomized complete block design with three replications at the Research Centre of Islamic Azad University of Firoozabad, Iran during 2010-2011 growing season. Eight drought tolerant indices mean productivity (MP), stress tolerance index (STI), geometric mean productivity (GMP), ...

  3. Tolerance

    DEFF Research Database (Denmark)

    Tønder, Lars

    Tolerance: A Sensorial Orientation to Politics is an experiment in re-orientation. The book is based on the wager that tolerance exceeds the more prevalent images of self-restraint and repressive benevolence because neither precludes the possibility of a more “active tolerance” motivated...... by the desire to experiment and to become otherwise. The objective is to discuss what gets lost, conceptually as well as politically, when we neglect the subsistence of active tolerance within other practices of tolerance, and to develop a theory of active tolerance in which tolerance's mobilizing character...... the current models of restraint and benevolence, other ways of understanding the politics of democratic pluralism might be developed, which will enable us to conceive of tolerance's future in terms different than those currently on offer. Tolerance: A Sensorial Orientation to Politics develops...

  4. Natural selection on individual variation in tolerance of gastrointestinal nematode infection.

    Science.gov (United States)

    Hayward, Adam D; Nussey, Daniel H; Wilson, Alastair J; Berenos, Camillo; Pilkington, Jill G; Watt, Kathryn A; Pemberton, Josephine M; Graham, Andrea L

    2014-07-01

    Hosts may mitigate the impact of parasites by two broad strategies: resistance, which limits parasite burden, and tolerance, which limits the fitness or health cost of increasing parasite burden. The degree and causes of variation in both resistance and tolerance are expected to influence host-parasite evolutionary and epidemiological dynamics and inform disease management, yet very little empirical work has addressed tolerance in wild vertebrates. Here, we applied random regression models to longitudinal data from an unmanaged population of Soay sheep to estimate individual tolerance, defined as the rate of decline in body weight with increasing burden of highly prevalent gastrointestinal nematode parasites. On average, individuals lost weight as parasite burden increased, but whereas some lost weight slowly as burden increased (exhibiting high tolerance), other individuals lost weight significantly more rapidly (exhibiting low tolerance). We then investigated associations between tolerance and fitness using selection gradients that accounted for selection on correlated traits, including body weight. We found evidence for positive phenotypic selection on tolerance: on average, individuals who lost weight more slowly with increasing parasite burden had higher lifetime breeding success. This variation did not have an additive genetic basis. These results reveal that selection on tolerance operates under natural conditions. They also support theoretical predictions for the erosion of additive genetic variance of traits under strong directional selection and fixation of genes conferring tolerance. Our findings provide the first evidence of selection on individual tolerance of infection in animals and suggest practical applications in animal and human disease management in the face of highly prevalent parasites.

  5. Natural selection on individual variation in tolerance of gastrointestinal nematode infection.

    Directory of Open Access Journals (Sweden)

    Adam D Hayward

    2014-07-01

    Full Text Available Hosts may mitigate the impact of parasites by two broad strategies: resistance, which limits parasite burden, and tolerance, which limits the fitness or health cost of increasing parasite burden. The degree and causes of variation in both resistance and tolerance are expected to influence host-parasite evolutionary and epidemiological dynamics and inform disease management, yet very little empirical work has addressed tolerance in wild vertebrates. Here, we applied random regression models to longitudinal data from an unmanaged population of Soay sheep to estimate individual tolerance, defined as the rate of decline in body weight with increasing burden of highly prevalent gastrointestinal nematode parasites. On average, individuals lost weight as parasite burden increased, but whereas some lost weight slowly as burden increased (exhibiting high tolerance, other individuals lost weight significantly more rapidly (exhibiting low tolerance. We then investigated associations between tolerance and fitness using selection gradients that accounted for selection on correlated traits, including body weight. We found evidence for positive phenotypic selection on tolerance: on average, individuals who lost weight more slowly with increasing parasite burden had higher lifetime breeding success. This variation did not have an additive genetic basis. These results reveal that selection on tolerance operates under natural conditions. They also support theoretical predictions for the erosion of additive genetic variance of traits under strong directional selection and fixation of genes conferring tolerance. Our findings provide the first evidence of selection on individual tolerance of infection in animals and suggest practical applications in animal and human disease management in the face of highly prevalent parasites.

  6. Selection criteria for drought tolerance at the vegetative phase in ...

    African Journals Online (AJOL)

    Aghomotsegin

    2016-05-18

    May 18, 2016 ... cultivated for its high economic importance as a food crop and an industrial raw material. .... and the fourth block till 35 DAP (WT35). For each water treatment, data were collected on percentage ..... Africa's Green revolution drought tolerance maize scam. Africa Centre for Biosafety Briefing Paper, no 12.

  7. Biomass equations for selected drought-tolerant eucalypts in South ...

    African Journals Online (AJOL)

    In the water-scarce environment of South Africa, drought-tolerant eucalypt species have the potential to contribute to the timber and biomass resource. Biomass functions are a necessary prerequisite to predict yield and carbon sequestration. In this study preliminary biomass models for Eucalyptus cladocalyx, ...

  8. A selective sweep in a microsporidian parasite Nosema-tolerant honeybee population, Apis mellifera

    DEFF Research Database (Denmark)

    Huang, Q.; Lattorff, H. M. G.; Kryger, P.

    2014-01-01

    Nosema is a microsporidian parasite of the honeybee, which infects the epithelial cells of the gut. In Denmark, honeybee colonies have been selectively bred for the absence of Nosema over decades, resulting in a breeding line that is tolerant toward Nosema infections. As the tolerance toward the ...

  9. 40 CFR 180.1108 - Delta endotoxin of Bacillus thuringiensis variety San Diego encapsulated into killed Pseudomonas...

    Science.gov (United States)

    2010-07-01

    ... thuringiensis variety San Diego encapsulated into killed Pseudomonas fluorescens; exemption from the requirement... into killed Pseudomonas fluorescens; exemption from the requirement of a tolerance. The delta endotoxin of Bacillus thuringiensis variety San Diego encapsulated into killed Pseudomonas fluorescens is...

  10. Functionally Selective Signaling for Morphine and Fentanyl Antinociception and Tolerance Mediated by the Rat Periaqueductal Gray

    Science.gov (United States)

    Morgan, Michael M.; Reid, Rachel A.; Saville, Kimber A.

    2014-01-01

    Functionally selective signaling appears to contribute to the variability in mechanisms that underlie tolerance to the antinociceptive effects of opioids. The present study tested this hypothesis by examining the contribution of G protein-coupled receptor kinase (GRK)/Protein kinase C (PKC) and C-Jun N-terminal kinase (JNK) activation on both the expression and development of tolerance to morphine and fentanyl microinjected into the ventrolateral periaqueductal gray of the rat. Microinjection of morphine or fentanyl into the periaqueductal gray produced a dose-dependent increase in hot plate latency. Microinjection of the non-specific GRK/PKC inhibitor Ro 32-0432 into the periaqueductal gray to block mu-opioid receptor phosphorylation enhanced the antinociceptive effect of morphine but had no effect on fentanyl antinociception. Microinjection of the JNK inhibitor SP600125 had no effect on morphine or fentanyl antinociception, but blocked the expression of tolerance to repeated morphine microinjections. In contrast, a microinjection of Ro 32-0432 blocked the expression of fentanyl, but not morphine tolerance. Repeated microinjections of Ro 32-0432 blocked the development of morphine tolerance and inhibited fentanyl antinociception whether rats were tolerant or not. Repeated microinjections of SP600125 into the periaqueductal gray blocked the development of tolerance to both morphine and fentanyl microinjections. These data demonstrate that the signaling molecules that contribute to tolerance vary depending on the opioid and methodology used to assess tolerance (expression vs. development of tolerance). This signaling difference is especially clear for the expression of tolerance in which JNK contributes to morphine tolerance and GRK/PKC contributes to fentanyl tolerance. PMID:25503060

  11. [Selection and characterization of salt-tolerant calli of Taraxacum officinale].

    Science.gov (United States)

    Zhang, Xinguo; Chen, Xianyang; Jiang, Dan; Li, Yinxin

    2008-07-01

    In order to obtain salt-tolerant calli of Dandelion (Taraxacum officinale Weber), calli were induced from leaf explants of Dandelion on Murashige and Skoog's medium supplemented with 2.0 mg/L 6-benzyladenine and 0.5 mg/L 2,4-dichlorophen oxyacetic acid With 1.5% NaCl as selection pressure, most calli became brown and dead, whereas some new cell clusters appeared at the edge of the brown calli after 2 to 3 weeks. The survived cells were picked out and sub-cultured every 3 weeks onto the fresh selection medium and salt-tolerant calli were finally obtained through 4 continuous selections on the selection medium supplemented with 1.5% NaCl. Salt-tolerant calli increased steadily under a fixed NaCl stress though their relative growth rate decreased with increased NaCl concentration whereas the control calli which were sub-cultured by 4 continus selections on salt free medium ceased to grow under the same condition. This result indicated that the salt-tolerance of the selected calli is improved and this character is stable. Compared with the control, the SDS-PAGE pattern of the salt-tolerant calli had a unique 34 kD protein band. Its 30 kD and 18 kD protein bands were up-regulated. Further more, within the NaCl stress range up to 1.5%, the activities of antioxidant enzymes such as super oxidase dimutase, peroxidase and catalase, and the proline contents of the salt-tolerant calli were higher than those of the control. The results indicated that the selected calli with improved and stable salt tolerance were cell variants. The accumulation of the organic compatible solutes including proteins and the enhanced antioxidant capabilities in the salt tolerant calli are the two ways for them to regulate their osmotic homeostasis and alleviate the secondary reactive oxygen spexies damage respectively.

  12. Selection of tolerable risk criteria for dam safety decision making

    International Nuclear Information System (INIS)

    Nielsen, N.M.; Hartford, D.N.D.; MacDonald, T.F.

    1994-01-01

    Risk assessment has received increasing attention in recent years as a means of aiding decision making on dams by providing systematic and rational methods for dealing with risk and uncertainty. Risk assessment is controversial and decisions affecting risk to life are the most controversial. Tolerable criteria, based on the risks that society is prepared to accept in order to avoid excessive costs, set bounds within which risk-based decisions may be made. The components of risk associated with dam safety are addressed on an individual basis and criteria established for each component, thereby permitting flexibility in the balance between component risk and avoiding the problems of placing a monetary value on life. The guiding principle of individual risk is that dams do not impose intolerable risks on any individual. A risk to life of 1 in 10 4 per annum is generally considered the maximum tolerable risk. When considering societal risk, the safety of a dam should be proportional to the consequences of its failure. Risks of financial losses beyond the corporation's ability to finance should be so low as to be considered negligible. 17 refs., 3 figs

  13. Improvement of potato tolerance to salinity using tissue culture techniques and irradiation with in vitro selection

    International Nuclear Information System (INIS)

    Al-Safadi, B.; Arabi, M. I. E.

    2005-06-01

    A mutation breeding program was conducted to improve potato (Solanum tuberosum) tolerance to salinity. In vitro cultured explants from potato cvs. Draga, Diamant, Spunta were irradiated with gamma doses 25, 30, and 35 Gy.Growing shoots were cut and re-cultured every 2 weeks until the 4th generation (MV 4 ) to make sure no chimeral tissues still existed in the mutant material. Plantlets were subsequently propagated to obtain enough explants for in vitro selection pressure. Around 3000 plantlets from the three cultivars were subjected to selection pressure. MV 4 explants were cultured on MS medium supplemented with the NaCl in varying concentrations ranging between 50 to 200 mM. Surviving plantlets were propagated and re-cultured on a similar medium to insure their tolerance to salinity. Tolerant plantlets were acclimatized and transferred to pots and grown under glasshouse conditions. Plants were later subjected to another selection pressure, by irrigating them using water containing NaCl in concentrations ranging between 50-250 mM in addition to controls irrigated with normal water. Cultivar Spunta produced the highest number of tolerant plants. Four plants of Spunta appeared to be tolerant to salinity whereas only one plant from Diamant and was tolerant and no plants from cultivar Draga were tolerant. Mutant plants varied in number of produced minitubers from 8 - 14. Also, weight of these minitubers varied from less than 1 to 31 grams. (author)

  14. Experiments on prevention of the endotoxin-abortifacient effect by radiodetoxified endotoxin pretreatment in rats

    Energy Technology Data Exchange (ETDEWEB)

    Csordas, T.; Bertok, L.; Csapo, Z.

    1978-01-01

    Endotoxemia has been induced in pregnant rats by intravenous injection of 1 mg Escherichia coli endotoxin which resulted in intrauterine death and abortion of fetuses in 24 h. The abortifacient effect of endotoxin, injected intravenously 24 h earlier. The authors suppose that the radiodetoxified endotoxin can be a good tool also in the prevention of human septic (endotoxin) shock in pregnancy.

  15. Selection of salt-tolerant variant of wheat in vitro

    International Nuclear Information System (INIS)

    Zheng, Q.; Zhu, Y.; Chen, W.; Tang, X.

    1990-01-01

    Full text: Growing-tip explants of four winter wheat varieties were cultured on basal medium supplemented with 2,4-D 2mg/l and various NaCI concentrations. The calli derived from 'Nongda 139' growing-tips were subcultured on the medium salinized with 0.4% NaCI for one year. Then they were cultured on a medium serially salinised with NaCI, increasing by 0.4% steps until 2%. Finally, surviving calli were transferred to a medium containing 0.4% NaCI for regeneration. 21 plants were obtained among which 3 were sterile. Dwarf, late ripening, shrunken grains and spike shape variations were observed, but they were non-heritable except dwarf and spike shape. Some potential NaCI-tolerant variants were identified. (author)

  16. Selection of pigs for improved coping with health and environmental challenges: breeding for resistance or tolerance?

    Directory of Open Access Journals (Sweden)

    Sarita Zhe Ying Guy

    2012-12-01

    Full Text Available The benefits of improved health and welfare in pigs have driven refinements in management and selection practices, one of which is the production of pig phenotypes that can maintain health and productivity by improving response against pathogens. Selection has traditionally been made for host resistance; but the alternative host defence mechanism – host tolerance – is now being considered, as breeding for disease tolerance allows maintenance of high performance across environments of increasing pathogenic load. A distinction must be made between these two mechanisms as they vary in their influence on host-pathogen interactions and pathogen evolution, and consequently on the results of breeding programmes. Many pig production studies have failed to distinguish between resistance and tolerance; although a distinction may not always be possible. This article reviews current perspectives in selective breeding for disease resistance and tolerance in growing pigs, and the attendant industry implications. To assess the viability of breeding for resistance and/or tolerance for improved response to disease and other environmental challenges, we propose the use of routine farm records, instead of data measurements taken from laboratory experiments. Consequently, a number of factors need to be taken into account simultaneously for a multidimensional modelling approach. This includes not only genotype and disease variables, but also descriptors of the environment, as well as any possible interactions. It may not be feasible to record individual pathogen loads, and therefore true tolerance, on farm using routinely collected data. However, it may be estimated with group (farm means, or other proxy measures. Although this results in a bias, this may still be useful for modelling and quantifying resistance and tolerance. We can then quantify success of selection, and this may enable us to decide whether to select for disease resistance versus disease

  17. Combined family selection for tolerance to corn reddening

    Directory of Open Access Journals (Sweden)

    Bekavac Goran

    2010-01-01

    Full Text Available Variability and interrelationships of traits in maize population NS 1-275 CRS after one cycle of combined HS-S1 family selection were studied. Independent field experiments with HS and S1 progenies were set up according to an incomplete block design in 2008 and 2009. The basic genetics parameters of zero (C0 and the first cycle of selection (C1 were assessed. The primary selection criterion for the formation of the C1 was grain yield (GY and grain moisture (GM and secondary stay green (SG and stalk lodging (SL. Significant differences in mean values between selection cycles (P£0.05 were determined for SG at S1 progenies and GY and SG at HS progenies. No significant changes in genetic variances were recorded after one cycle of selection. Positive correlations between GY and SG and negative between SG and SL exclude the occurrence of negative correlative response. In the upcoming period, some other agronomic traits have to be analyzed to get complete picture of their complex interrelationships, as well as the expected changes through cycles of selection. .

  18. Faecal Endotoxin in Human Volunteers: Normal Values

    OpenAIRE

    Van Saene, J. J. M.; Stoutenbeek, C. P.; Van Saene, H. K. F.

    2011-01-01

    This paper presents a study measuring endotoxin levels in faeces of healthy adults using the Limulus amoebocyte lysate (LAL) microassay. Data showed mean faecal endotoxin levels of 1 mg (range 0.01-100 mg) per gram of faeces. No correlation was found with the concentration of Enterobacteriaceae in faeces and free Faecal endotoxin levels.Keywords: Endotoxin; LAL micro-assay; LPS; Faeccs; Human volunteers: Enterobacteriaceae.

  19. 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

  20. Capacitive biosensor for detection of endotoxin.

    Science.gov (United States)

    Limbut, Warakorn; Hedström, Martin; Thavarungkul, Panote; Kanatharana, Proespichaya; Mattiasson, Bo

    2007-09-01

    A capacitive biosensor for the detection of bacterial endotoxin has been developed. Endotoxin-neutralizing protein derived from American horseshoe crab was immobilized to a self-assembled thiol layer on a biosensor transducer (Au). Upon injection of a sample containing endotoxin, a decrease in the observed capacitive signal was registered. Endotoxin could be determined under optimum conditions with a detection limit of 1.0 x 10(-13) M and linearity ranging from 1.0 x 10(-13) to 1.0 x 10(-10) M. Good agreement was achieved when applying endotoxin preparations purified from an Escherichia coli cultivation to the capacitive biosensor system, utilizing the conventional method for quantitative endotoxin determination, the Limulus amebocyte lysate test as a reference. The capacitive biosensor method was statistically tested with the Wilcoxon signed rank test, which proved the system is acceptable for the quantitative analysis of bacterial endotoxin (P<0.05).

  1. Selection of arabica coffee progenies tolerant to heat stress

    Directory of Open Access Journals (Sweden)

    Alexsandro Lara Teixeira

    2015-07-01

    Full Text Available Due to high temperatures, practically all coffee farms in the state of Rondonia are of the C. canephora species. Thus, importing arabica coffee from other states becomes necessary for composition of blends, as well as for the specialty or gourmet coffee market. The purpose of this study was to select arabica coffee genotypes that exhibit satisfactory agronomic performance under high temperature conditions. The experiment was conducted in OuroPreto do Oeste, RO, Brazil, with mean annual temperature of 25.8°C and mean annual rainfall of 2300mm year-1. The experiment was composed of 114 arabica coffee genotypes, with 103 progenies and eleven control cultivars, provided by EPAMIG. A randomized block experimental design was used with three replications, spacing of 3.0x1.0 meters and five plants per plot. All the crop seasons showed significant difference for the green coffee yield trait. In joint analysis, significant differences were detected among progenies and control cultivars. In the average of the four harvests, green coffee yield was 32.38 bags ha-1. The cultivars 'CatuaíVermelho IAC 15', 'Obatã IAC 1669-20' and 'Catucaí Amarelo 2SLCAK' stood out, achieving yields greater than 40 bags ha-1. The gain obtained from selection was 14.33 bags ha-1, which is equivalent to an increase of 44.04% in production of green coffee. The progeny H514-7-10-6-2-3-9 stood out with an average yield of 51.20 bags ha-1. In regard to maturation cycle, 56% of the progenies were classified as early maturity and 44% as medium maturity. Late maturity genotypes were not observed

  2. Population structure, genetic variation and linkage disequilibrium in perennial ryegrass populations divergently selected for freezing tolerance

    Directory of Open Access Journals (Sweden)

    Mallikarjuna Rao eKovi

    2015-11-01

    Full Text Available Low temperature is one of the abiotic stresses seriously affecting the growth of perennial ryegrass (Lolium perenne L. Understanding the genetic control of freezing tolerance would aid in the development of cultivars of perennial ryegrass with improved adaptation to frost. A total number of 80 individuals (24 of High frost [HF]; 29 of Low frost [LF] and 27 of Unselected [US] from the second generation of the two divergently selected populations and an unselected control population were genotyped using 278 genome-wide SNPs derived from Lolium perenne L. transcriptome sequence. Our studies showed that the HF and LF populations are very divergent after selection for freezing tolerance, whereas the HF and US populations are more similar. Linkage disequilibrium (LD decay varied across the seven chromosomes and the conspicuous pattern of LD between the HF and LF population confirmed their divergence in freezing tolerance. Furthermore, two Fst outlier methods; finite island model (fdist by LOSITAN and hierarchical structure model using ARLEQUIN detected six loci under directional selection. These outlier loci are most probably linked to genes involved in freezing tolerance, cold adaptation and abiotic stress and might be the potential marker resources for breeding perennial ryegrass cultivars with improved freezing tolerance.

  3. Mechanistic Insight into Salt Tolerance of Acacia auriculiformis: The Importance of Ion Selectivity, Osmoprotection, Tissue Tolerance, and Na+ Exclusion

    Science.gov (United States)

    Rahman, Md. M.; Rahman, Md. A.; Miah, Md. G.; Saha, Satya R.; Karim, M. A.; Mostofa, Mohammad G.

    2017-01-01

    Salinity, one of the major environmental constraints, threatens soil health and consequently agricultural productivity worldwide. Acacia auriculiformis, being a halophyte, offers diverse benefits against soil salinity; however, the defense mechanisms underlying salt-tolerant capacity in A. auriculiformis are still elusive. In this study, we aimed to elucidate mechanisms regulating the adaptability of the multi-purpose perennial species A. auriculiformis to salt stress. The growth, ion homeostasis, osmoprotection, tissue tolerance and Na+ exclusion, and anatomical adjustments of A. auriculiformis grown in varied doses of seawater for 90 and 150 days were assessed. Results showed that diluted seawater caused notable reductions in the level of growth-related parameters, relative water content, stomatal conductance, photosynthetic pigments, proteins, and carbohydrates in dose- and time-dependent manners. However, the percent reduction of these parameters did not exceed 50% of those of control plants. Na+ contents in phyllodes and roots increased with increasing levels of salinity, whereas K+ contents and K+/Na+ ratio decreased significantly in comparison with control plants. A. auriculiformis retained more Na+ in the roots and maintained higher levels of K+, Ca2+ and Mg2+, and K+/Na+ ratio in phyllodes than roots through ion selective capacity. The contents of proline, total free amino acids, total sugars and reducing sugars significantly accumulated together with the levels of malondialdehyde and electrolyte leakage in the phyllodes, particularly at day 150th of salt treatment. Anatomical investigations revealed various anatomical changes in the tissues of phyllodes, stems and roots by salt stress, such as increase in the size of spongy parenchyma of phyllodes, endodermal thickness of stems and roots, and the diameter of root vascular bundle, relative to control counterparts. Furthermore, the estimated values for Na+ exclusion and tissue tolerance index suggested that

  4. Resistance and Tolerance to Tropodithietic Acid, an Antimicrobial in Aquaculture, Is Hard To Select

    DEFF Research Database (Denmark)

    Porsby, Cisse Hedegaard; Webber, Mark A.; Nielsen, Kristian Fog

    2011-01-01

    of development of TDA resistance. A bacterial extract containing 95% TDA was effective against a range of human-pathogenic bacteria, including both Gram-negative and Gram-positive bacteria. TDA was bactericidal against Salmonella enterica serovar Typhimurium SL1344 and Staphylococcus aureus NCTC 12493 and killed...... mutants, or prolonged exposure to incremental concentrations of TDA) resulted in resistant or tolerant strains. After more than 300 generations exposed to sub-MIC and MIC concentrations of a TDA-containing extract, strains tolerant to 2x the MIC of TDA for wild-type strains were selected...... both growing and nongrowing cells. Several experimental approaches were used to select mutants resistant to TDA or subpopulations of strains with enhanced tolerance to TDA. No approach (single exposures to TDA extract administered via different methods, screening of a transposon library for resistant...

  5. [Selection and identification of salt-tolerant variants of Taraxacum officinale].

    Science.gov (United States)

    Zhang, Xinguo; Li, Yinxin; Chen, Hua; Shi, Wuliang

    2008-02-01

    In order to obtain salt-tolerant variant plants of Dandelion (Taraxacum officinale Weber), the leaf discs were excised from 20 to 30-day old seedlings to produce callus, then the induced calli were transferred to selection mediums containing 1.5% NaCl. After regenerating and rooting, these salt-tolerant calli finally developed into 12 variant plantlets. Compared with the wild-type, these regenerated plants produced more trichomes on their leaves, and had larger leaves and shorter petioles. Additionally, the dumpy roots and an approximately 2-cm bract in middle parts of the floricanes were clearly observed in these salt-tolerant plants. By RAPD (Random Amplified Polymorphic DNA) and SDS-PAGE analysis, these salt-tolerant plants showed differences from the control at DNA and protein levels. With 1.5% NaCl treatment, the antioxidant enzyme activity, proline content, and flavonoid concentration were higher in these salt-tolerant plants, whereas maloaldehyde concentration was significantly lower. Salt-tolerant lines of T. officinale showed stronger anti-oxidative activity and higher flavonoid contents.

  6. In vitro application of integrated selection index for screening drought tolerant genotypes in common wheat

    Directory of Open Access Journals (Sweden)

    Ezatollah FARSHADFAR

    2016-10-01

    Full Text Available This experiment was conducted on 20 wheat genotypes during 2010-2011 growing season at the Razi University, Kermanshah, Iran. A completely randomized design with six replications was used for callus induction and a 20 × 2 factorial experiment with three replications was used for response of genotypes to in vitro drought stress. ANOVA exhibited highly significant differences among the genotypes for callus growth rate, relative fresh mass growth, relative growth rate, callus water content, percent of callus chlorosis and proline content under stress condition (15 % PEG. PCA showed that the integrated selection index was correlated with callus growth index, relative fresh mass growth, relative growth rate and proline content indicating that these screening techniques can be useful for selecting drought tolerant genotypes. Screening drought tolerant genotypes and in vitro indicators of drought tolerance using mean rank, standard deviation of ranks and biplot analysis, discriminated genotypes 2, 18 and 10 as the most drought tolerant. Therefore they are recommended to be used as parents for genetic analysis, gene mapping and improvement of drought tolerance.

  7. In vitro selection of mutants: Inducible gene regulation for salt tolerance

    International Nuclear Information System (INIS)

    Winicov, I.; Bastola, D.R.; Deutch, C.E.; Pethe, V.V.; Petrusa, L.

    2001-01-01

    Regulation of differentially expressed genes in plants may be involved in inducing tolerance to stress. Isogenic salt-sensitive and salt-tolerant alfalfa lines were investigated for molecular differences in their response to salt. The genes, which are differentially induced by salt in the salt-tolerant alfalfa cells and are also regulated by salt at the whole plant level, were cloned. Both transcriptional and post- transcriptional mechanisms influenced salt-induced product accumulation in the salt-tolerant alfalfa. The salt-tolerant plants doubled proline concentration rapidly in roots, while salt-sensitive plants showed a delayed response. To understand the regulatory system in the salt-tolerant alfalfa, two genes that are expressed in roots were studied. Alfin1 encodes a zinc-finger type putative DNA transcription factor conserved in alfalfa, rice and Arabidopsis, and MsPRP2 encodes a protein that serves as a cell wall- membrane linker in roots. Recombinant Alfin1 protein was selected, amplified, cloned and its consensus sequence was identified. The recombinant Alfin1 also bound specifically to fragments of the MsPRP2 promoter in vitro, containing the Alfin1 binding consensus sequence. The results show unambiguously binding specificity of Alfin1 DNA, supporting its role in gene regulation. Alfin1 function was tested in transformed alfalfa in vivo by over-expressing Alfin1 from 35S CaMV promoter. The transgenic plants appeared normal. However, plants harboring the anti-sense construct did not grow well in soil, indicating that Alfin1 expression was essential. Alfin1 over-expression in transgenic alfalfa led to enhanced levels of MsPRP2 transcript accumulation, demonstrating that Alfin1 functioned in vivo in gene regulation. Since MsPRP2 gene is also induced by salt, it is likely that Alfin1 is an important transcription factor for gene regulation in salt-tolerant alfalfa, and an excellent target for manipulation to improve salt tolerance. (author)

  8. Recent advances in biosensor based endotoxin detection.

    Science.gov (United States)

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

    2014-01-15

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

  9. Proanthocyanidin-based Endotoxin Removal

    Science.gov (United States)

    2014-01-16

    synthesis or purification of a variety of compounds with LPS-binding activities [2–11]. Polymyxin B (PMB) is the most commonly employed agent for removal... polyphenolics are accessible to the reagents in a manner similar to those in free in solution. The analysis also assumes that all degrees of... Synthesis and Application of a Novel Ligand for Affinity Chromatography Based Removal of Endotoxin from Antibodies,” Bioconjugate Chemistry 18, 559

  10. Endotoxins in surgical instruments of hip arthroplasty.

    Science.gov (United States)

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

    2016-01-01

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

  11. Breeding approaches in simultaneous selection for multiple stress tolerance of maize in tropical environments

    Directory of Open Access Journals (Sweden)

    Denić M.

    2007-01-01

    Full Text Available Maize is the principal crop and major staple food in the most countries of Sub-Saharan Africa. However, due to the influence of abiotic and biotic stress factors, maize production faces serious constraints. Among the agro-ecological conditions, the main constraints are: lack and poor distribution of rainfall; low soil fertility; diseases (maize streak virus, downy mildew, leaf blights, rusts, gray leaf spot, stem/cob rots and pests (borers and storage pests. Among the socio-economic production constraints are: poor economy, serious shortage of trained manpower; insufficient management expertise, lack of use of improved varieties and poor cultivation practices. To develop desirable varieties, and thus consequently alleviate some of these constraints, appropriate breeding approaches and field-based methodologies in selection for multiple stress tolerance, were implemented. These approaches are mainly based on: a Crossing selected genotypes with more desirable stress tolerant and other agronomic traits; b Using the disease/pest spreader row method, combined with testing and selection of created progenies under strong to intermediate pressure of drought and low soil fertility in nurseries; and c Evaluation of the varieties developed in multi-location trials under low and "normal" inputs. These approaches provide testing and selection of large number of progenies, which is required for simultaneous selection for multiple stress tolerance. Data obtained revealed that remarkable improvement of the traits under selection was achieved. Biggest progress was obtained in selection for maize streak virus and downy mildew resistance, flintiness and earliness. In the case of drought stress, statistical analyses revealed significant negative correlation between yield and anthesis-silking interval, and between yield and days to silk, but positive correlation between yield and grain weight per ear.

  12. Legionella Persistence in Manufactured Water Systems: Pasteurization Potentially Selecting for Thermal Tolerance

    Directory of Open Access Journals (Sweden)

    Harriet Whiley

    2017-07-01

    Full Text Available Legionella is an opportunistic waterborne pathogen of increasing public health significance. Pasteurization, otherwise known as super-heat and flush (increasing water temperature to above 70°C and flushing all outlets, has been identified as an important mechanism for the disinfection of Legionella in manufactured water systems. However, several studies have reported that this procedure was ineffective at remediating water distribution systems as Legionella was able to maintain long term persistent contamination. Up to 25% of L. pneumophila cells survived heat treatment of 70°C, but all of these were in a viable but non-culturable state. This demonstrates the limitations of the culture method of Legionella detection currently used to evaluate disinfection protocols. In addition, it has been demonstrated that pasteurization and nutrient starvation can select for thermal tolerant strains, where L. pneumophila was consistently identified as having greater thermal tolerance compared to other Legionella species. This review demonstrates that further research is needed to investigate the effectiveness of pasteurization as a disinfection method. In particular, it focuses on the potential for pasteurization to select for thermal tolerant L. pneumophila strains which, as the primary causative agent of Legionnaires disease, have greater public health significance compared to other Legionella species.

  13. Strategy for selection of soybean genotypes tolerant to drought during germination.

    Science.gov (United States)

    Dantas, S A G; Silva, F C S; Silva, L J; Silva, F L

    2017-05-10

    Water deficit is the main reason for instability in the context of soybean culture. The development of strategies for the selection of more tolerant genotypes is necessary. These strategies include the use of polyethylene glycol 6000 solutions (PEG-6000) for conducting the germination test under conditions of water restriction. Thus, the objective of this study was to determine the osmotic potential and the main characteristics that promote the discrimination of soybean genotypes with regard to water stress tolerance during germination and the vigor test. Thirteen soybean cultivars were used. The seeds were allowed to germinate on sheets of germitest paper moistened in solution with PEG-6000, simulating different levels of water availability, which is expressed as osmotic potential (0.0, -0.2, -0.4, and -0.6 MPa). We assessed germination, length, and dry mass for seedlings and seeds, as well as reserve dynamics. Germination and variables related to the dynamics of reservation have great influence on the expression of variability in environments under stress. Among the different osmotic potentials, the -0.2 MPa was the most efficient for the expression of genetic variability among the cultivars. Conducting the germination test with PEG-6000 solution to -0.2 MPa was efficient for selecting soybean cultivars tolerant to water stress. This was accomplished by evaluating the percentage of germination, along with variables related to the dynamics of reservation.

  14. Selective proteomic analysis of antibiotic-tolerant cellular subpopulations in pseudomonas aeruginosa biofilms

    DEFF Research Database (Denmark)

    Babin, Brett M.; Atangcho, Lydia; van Eldijk, Mark B.

    2017-01-01

    to determine the dynamic proteomic response of the tolerant subpopulation to supra-MIC treatment with the fluoroquinolone antibiotic ciprofloxacin. The adaptive response included the upregulation of proteins required for sensing and repairing DNA damage and substantial changes in the expression of enzymes...... involved in central carbon metabolism. We differentiated the immediate proteomic response, characterized by an increase in flagellar motility, from the long-term adaptive strategy, which included the upregulation of purine synthesis. This targeted, selective analysis of a bacterial subpopulation......, in which distinct cellular subpopulations can respond differently to stresses, including subpopulations of pathogenic biofilms that are more tolerant to antibiotics. Global proteomic analysis affords insights into cellular physiology but cannot identify proteins expressed in a particular subpopulation...

  15. K-selection, α-selection, effectiveness, and tolerance in competition ...

    Indian Academy of Sciences (India)

    We also use this broader formulation to predict that selection for faster development in Drosophila should, in fact, lead to the correlated evolution of decreased competitive ability, even though it does lead to the evolution of greater efficiency and higher population growth rates at high density when in monotypic culture.

  16. K-selection, α-selection, effectiveness, and tolerance in competition ...

    Indian Academy of Sciences (India)

    Unknown

    In this formulation, N represents the total population size, regardless of genotype, whereas the r and K terms are genotype specific; consequently, these models have often been referred to as pure density-dependent selection models because there are no frequency-dependent inter- genotypic interactions. At the time these ...

  17. Genomic Selection for Drought Tolerance Using Genome-Wide SNPs in Maize

    Directory of Open Access Journals (Sweden)

    Thirunavukkarasu Nepolean

    2017-04-01

    Full Text Available Traditional breeding strategies for selecting superior genotypes depending on phenotypic traits have proven to be of limited success, as this direct selection is hindered by low heritability, genetic interactions such as epistasis, environmental-genotype interactions, and polygenic effects. With the advent of new genomic tools, breeders have paved a way for selecting superior breeds. Genomic selection (GS has emerged as one of the most important approaches for predicting genotype performance. Here, we tested the breeding values of 240 maize subtropical lines phenotyped for drought at different environments using 29,619 cured SNPs. Prediction accuracies of seven genomic selection models (ridge regression, LASSO, elastic net, random forest, reproducing kernel Hilbert space, Bayes A and Bayes B were tested for their agronomic traits. Though prediction accuracies of Bayes B, Bayes A and RKHS were comparable, Bayes B outperformed the other models by predicting highest Pearson correlation coefficient in all three environments. From Bayes B, a set of the top 1053 significant SNPs with higher marker effects was selected across all datasets to validate the genes and QTLs. Out of these 1053 SNPs, 77 SNPs associated with 10 drought-responsive transcription factors. These transcription factors were associated with different physiological and molecular functions (stomatal closure, root development, hormonal signaling and photosynthesis. Of several models, Bayes B has been shown to have the highest level of prediction accuracy for our data sets. Our experiments also highlighted several SNPs based on their performance and relative importance to drought tolerance. The result of our experiments is important for the selection of superior genotypes and candidate genes for breeding drought-tolerant maize hybrids.

  18. Airborne endotoxin concentrations in various work areas within two cotton textile mills in the People's Republic of China.

    Science.gov (United States)

    Olenchock, S A; Christiani, D C; Mull, J C; Ye, T T; Lu, P L

    1990-12-01

    As part of a multidisciplinary longitudinal approach to assess the roles of airborne cotton dust and endotoxins in affecting the respiratory health of cotton textile workers, this study was designed to quantify the endotoxin contamination of airborne vertically elutriated and total dusts. Yarn preparation areas (opening through fine spinning) were studied at two cotton textile mills which had been studied 5 years previously in Shanghai. People's Republic of China, Filter, with vertically elutriated (VE) or total dusts were mailed to the United States and endotoxin analyses were performed for each filter in duplicate with the quantitative chromogenic modification of the Limulus amebocyte lysate assay. Dusts from all areas of the textile mills contained endotoxins. Endotoxin burdens in VE dusts from the carding area were similar in both milk while the endotoxin contamination of total dust from carding in Mill 1 was over threefold greater than that of total dust from carding in Mill 2. All other areas differed between milk in both VE and total dust endotoxin burdens. Mean endotoxin levels in VE dusts from all areas of both mills were well above the reported threshold of 90 EU/m3 for acute pulmonary function effects in humans. Comparison of selected areas of both mills from the present study with the same work areas from the previous study showed that, in general, the airborne endotoxin burden was higher than levels found 5 years ago in these two mills. The data suggest that even with reduced or unchanged gravimetric dust levels in these two cotton textile mills, airborne endotoxin levels were higher and provided an increased potential for adverse respiratory response in exposed workers.

  19. Selection of Rice Genotypes for Salinity Tolerance Through Morpho-Biochemical Assessment

    Directory of Open Access Journals (Sweden)

    Md. Nasim ALI

    2014-09-01

    Full Text Available The present study reported the morpho-biochemical evaluation of 15 selected rice genotypes for salt tolerance at the seedling stage. Growth parameters including shoot length, root length, plant biomass, plant turgid weight, plant dry weight along with relative water content were measured after exposure to saline solution (with electrical conductivity value of 12 dS/m. Genotypes, showing significant differential responses towards salinity in the fields, were assessed through 14 salinity-linked morpho-biochemical attributes, measured at 14 d after exposure of seedling in saline nutrient solution. Relative water content, chlorophyll a/b, peroxidase activity and plant biomass were identified as potential indicators of salt tolerance. Principal component analysis and successive Hierarchical clustering using Euclidean distance revealed that Talmugur, Gheus, Ghunsi, Langalmura, Sabitapalui, and Sholerpona were promising genotypes for further breeding programmes in rice. The maximum Euclidean distance was plotted between Thavallakanan and Talmugur (7.49, followed by Thavallakanan and Langalmura (6.82, indicating these combinations may be exploited as parental lines in hybridization programmes to develop salinity tolerant variety.

  20. 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.

  1. Characterization and Selection of Phosphorus Deficiency Tolerant Rice Genotypes in Sri Lanka

    Directory of Open Access Journals (Sweden)

    Y.C. Aluwihare

    2016-07-01

    Full Text Available Phosphorus (P deficiency in soil is a major constrain for rice production. An important set of rice genotypes (landraces, old improved and new improved varieties were screened for P deficiency tolerance in two major cropping seasons of Sri Lanka, in 2012. The Ultisol soil, which was collected from a plot cultivated with rice without fertilizer application for past 40 years (P0 at the Rice Research and Development Institute (RRDI, Bathalagoda, Sri Lanka, was used as the potting medium for greenhouse trials. Two field trials were conducted in the same plots at RRDI. Both P0 and P30 (30 mg/kg P2O5 conditions were used in the two greenhouse trials. At the early vegetative (three weeks after transplanting, late vegetative (six weeks after transplanting and flowering stages, plant height and number of tillers per plant were recorded. At the flowering stage, shoots were harvested and shoot dry weight, shoot P concentration, shoot P uptake and P utilization efficiency were measured. All data were statistically analyzed using analysis of variance, regression and cluster procedures. The measured parameters were significantly different between P0 and P30 conditions (P < 0.05. Higher shoot dry weight was reported by the rice genotypes H4 and Marss under P0 conditions. The regression analysis between shoot dry weight and P utilization efficiency revealed that the studied rice genotypes could be categorized to three P deficiency tolerance classes. A total of 13 genotypes could be considered as highly tolerant and 4 genotypes as sensitive for P deficiency. These results could be used to select parental genotypes for breeding and genetic studies and also to select interesting varieties or landraces for organic rice production.

  2. Selective immediate responders to amoxicillin and clavulanic acid tolerate penicillin derivative administration after confirming the diagnosis.

    Science.gov (United States)

    Blanca-Lopez, N; Perez-Alzate, D; Ruano, F; Garcimartin, M; de la Torre, V; Mayorga, C; Somoza, M L; Perkins, J; Blanca, M; Canto, M G; Torres, M J

    2015-08-01

    An increasing number of patients show immediate selective hypersensitivity reactions to clavulanic acid (CLV) and amoxicillin (AX), probably due to their increased prescription. The maintenance of this response should be established. To assess that the immediate hypersensitivity selective response to AX or to CLV is maintained after repeated administration of penicillin G (PG)/penicillin V (PV) and AX. Patients with proven immediate hypersensitivity to AX (Group A) or CLV (Group B) were included. Diagnosis was performed using skin tests with major and minor determinants of PG (PPL/MDM), AX and CLV and by drug provocation test (DPT) if required. Selectivity was established by confirming tolerance to PG/PV (Group A) and to PG/PV and AX (Group B). The maintenance of the selective response was verified by repeating DPT, 15 days after the initial investigation, with the same procedure. Of 51 patients, 78% belonged to Group A and 22% to Group B. Most had anaphylaxis. In Group A, 72% were skin test positive; 28% required DPT. In Group B, 63% were skin test positive; 37% required DPT. Only two AX-selective cases developed positive responses after re-provocation with PG/PV. No cases selective for CLV developed a positive response to PG, PV or AX. The selective response to AX appears consistent, and a response to penicillin determinants only develops in a minority of cases. For the case of CLV, the selective response appears not to be modified by exposure to penicillin determinants, meaning that patients with CLV allergy can take penicillin derivatives safely. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  3. Selection of rice mutants Oryza Sativa L. with tolerance to saline grounds

    International Nuclear Information System (INIS)

    Hernandez Aguero, L.A.

    2001-01-01

    A selection of rice mutants with tolerance to salinity, took place in the Escuela de Ciencias Agrarias de la Universidad Nacional de Heredia, in conditions of hothouse starting from a population of M2 segregative seed, coming from commercial seed radiated with Co 60 gamma rays. The studied segregatives were: Setesa-9, Experimental II and Experimental I. For making this selection, the seed M2 was planted in plastic trays with saline soil with electrical conductivity values from 8 to 10 mmhos/cm. In each case, non-radiated original seed was used as control. After 22 days the seedling germinated, an evaluation was made and it was seen that any of the controls had resisted to the saline stress, and only those segregatives resistent to salinity survived. These were the next ones: 9 individuals of Setesa, 10 of the Experimental II, and 9 of Experimental I. The index of selection obtained was: 3.6, 4.0 y 3.6 respectively. In a second phase of the experiment, the seedling selected as salinity resistant, were taken to the ground were they were developed for getting the M3 mutant seed tolerant to salinity. The plants were individually harvested in the ground and each one had a specific identification. Then, weight and number data, fertile grain and ineffectives of the M3 seed were taken. After, for corroborating the capacity of tolerance to salinity, M3 seed was planted in flowerpots with saline soil with a value of electrical conductivity between 8 and 10 mmhos/cm. After data were analyzed, it was proved that some rice mutants had a profit of even 28 grams for 1000 grams as: ExpI-17, ExpI-15, ExpI-08, ExpII-22, ExpII-08, ExpII-30 and Se-9-14, Se-9-39 and Se-9-10. Therefore, the methodology utilized showed being effective and efficient for the objectives of the work [es

  4. Hypo-responsiveness of interleukin-8 production in human embryonic epithelial intestine 407 cells independent of NF-κB pathway: New lessons from endotoxin and ribotoxic deoxynivalenol

    International Nuclear Information System (INIS)

    Moon, Yuseok; Yang, Hyun; Park, Seung-Hwan

    2008-01-01

    Mucosal epithelium senses external toxic insults and transmits the danger signals into the epithelial cells in order to activate a broad range of inflammatory responses. However, pre-exposure to the commensal endotoxins can induce inflammatory tolerance and maintain the homeostasis without excessive immune responses. We recently reported that ribotoxin deoxynivalenol (DON) and its derivatives elicited the pro-inflammatory response as the mucosal insults in human epithelial cells. Taking the knowledge into consideration, we tested the hypothesis that endotoxin pre-exposure can attenuate ribotoxin-induced epithelial interleukin-8 (IL-8) production via a tolerance mechanism. Pre-exposure to endotoxin repressed IL-8 release and its gene expression. However, inflammatory tolerance was not mediated by the attenuated NF-κB activation which has been generally recognized as the major mediator of LPS-mediated toll-like receptor (TLR) signaling pathway. Instead, pre-exposure to endotoxin was observed to trigger the delayed induction of peroxisome proliferator-activated receptor gamma (PPAR-γ) which contributed to the diminished IL-8 production in the human epithelial cells. Moreover, endogenous PPAR-γ agonist suppressed toxicant-mediated interleukin-8 production and IL-8 mRNA stability. Taken together, endotoxin induced hypo-production of pro-inflammatory cytokine IL-8 in the human epithelial cells, which was associated with the delayed activation of PPAR-γ expression by pre-existing endotoxin

  5. In-vitro selection of plants resistant or tolerant to pathogenic fungi

    International Nuclear Information System (INIS)

    Dutrecq, A.J.E.

    1977-01-01

    A technique was developed to test the inhibition of germination of barley seeds by culture filtrates of Helminthosporium sativum P., K and B. Seeds were soaked for 24h in toxin preparations, and then placed for 48h in wet sand. Using a log-probit scale, a quasilinear relationship was found between seed germination and toxin concentration. The toxic effect was markedly affected by the pH of culture filtrates, being maximum at pH 2.0. Control pH 2.0 filtrates of uninoculated medium did not affect the germination of barley seeds under similar condition. Culture filtrates of H. sativum were also tested as inhibitors of multiplication of barley cells in suspensions. The purified toxin helminthosporal isolated from culture filtrates of H. sativum inhibited rhizogenesis in wheat calluses obtained from tissue culture. The techniques described are being used for the selection of cereals tolerant to H. sativum toxin. (author)

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

    Science.gov (United States)

    Meitern, Richard; Sild, Elin; Lind, Mari-Ann; Männiste, Marju; Sepp, Tuul; Karu, Ulvi; Hõrak, Peeter

    2013-01-01

    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. PMID:23805316

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

    Science.gov (United States)

    Meitern, Richard; Sild, Elin; Lind, Mari-Ann; Männiste, Marju; Sepp, Tuul; Karu, Ulvi; Hõrak, Peeter

    2013-01-01

    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.

  9. Selective labelling and eradication of antibiotic-tolerant bacterial populations in Pseudomonas aeruginosa biofilms

    DEFF Research Database (Denmark)

    Chua, Song Lin; Yam, Joey Kuok Hoong; Hao, Piliang

    2016-01-01

    subpopulations, with colistin-tolerant cells using type IV pili to migrate onto the top of the colistin-killed biofilm. The colistin-tolerant cells employ quorum sensing (QS) to initiate the formation of new colistin-tolerant subpopulations, highlighting multicellular behaviour in antibiotic tolerance......Drug resistance and tolerance greatly diminish the therapeutic potential of antibiotics against pathogens. Antibiotic tolerance by bacterial biofilms often leads to persistent infections, but its mechanisms are unclear. Here we use a proteomics approach, pulsed stable isotope labelling with amino...... acids (pulsed-SILAC), to quantify newly expressed proteins in colistin-tolerant subpopulations of Pseudomonas aeruginosa biofilms (colistin is a 'last-resort' antibiotic against multidrug-resistant Gram-negative pathogens). Migration is essential for the formation of colistin-tolerant biofilm...

  10. Elucidating the role of genetic drift and natural selection in cork oak differentiation regarding drought tolerance.

    Science.gov (United States)

    Ramírez-Valiente, J A; Lorenzo, Z; Soto, A; Valladares, F; Gil, L; Aranda, I

    2009-09-01

    Drought is the main selection agent in Mediterranean ecosystems and it has been suggested as an important evolutionary force responsible for population diversification in these types of environments. However, population divergence in quantitative traits can be driven by either natural selection, genetic drift or both. To investigate the roles of these forces on among-population divergence in ecophysiological traits related to drought tolerance (carbon isotope discrimination, specific leaf area, leaf size and leaf nitrogen content), we compared molecular and quantitative genetic differentiation in a common garden experiment including thirteen cork oak (Quercus suber L.) populations across a gradient of rainfall and temperature. Population differentiation for height, specific leaf area, leaf size and nitrogen leaf content measured during a dry year far exceeded the molecular differentiation measured by six nuclear microsatellites. Populations from dry-cool sites showed the lowest nitrogen leaf content and the smallest and thickest leaves contrasting with those from humid-warm sites. These results suggest (i) these traits are subjected to divergence selection and (ii) the genetic differences among populations are partly due to climate adaptation. By contrast, the low among-population divergence found in basal diameter, annual growth and carbon isotopic discrimination (a surrogate for water use efficiency) suggests low or no divergence selection for these traits. Among-population differentiation for neutral markers was not a good predictor for differentiation regarding the quantitative traits studied here, except for leaf size. The correlation observed between the genetic differentiation for leaf size and that for molecular markers was exclusively due to the association between leaf size and the microsatellite QpZAG46, which suggests a possible linkage between QpZAG46 and genes encoding for leaf size.

  11. Selection of Almond Vegetative Rootstocks for Water Stress Tolerance Based on the Morphological Markers

    Directory of Open Access Journals (Sweden)

    A.A. Shokouhian

    2016-02-01

    Full Text Available Introduction: One of the microbiological preparations used for this study was Effective Microorganisms (EM, being a commercial mixture of photosynthesizing bacteria, Actinomycetes, lactic acid bacteria, yeasts and fermenting fungi. The microbiological composition of the EM concentrateincludesStreptomyces albus, Propioni bacterium freudenreichil, Streptococcus lactis, Aspergillus oryzae, Mucor hiemalis, Saccharomycescerevisiae and Candida utilis. Moreover, EM also contains an unspecified amount of Lactobacillus sp. Rhodo pseudomonas sp. and Streptomyces griseus. Effective Microorganisms have a positive effect on the decomposition of organic matter, limiting putrefaction, increasing nitrogen content in the root medium of plants, phosphorus, improving soil fertility and as a result contributing to the growth and development of the root systems of plants. Selection of almond vegetative rootstocks for water stress tolerance is important for almond crop production in arid and semi-arid regions. The study of the eco-morphological characteristics that determine the success of a rootstock in a particular environment is a powerful tool for both agricultural management and breeding purposes. The aim of this work was to select the new rootstocks for water shortage tolerance, impact of water stress as well as Effective Microorganism (EM on morphological characteristics of almond rootstocks. Materials and Methods: In order to select the new rootstocks for water shortage tolerance, impact of water stress as well as EMonmorphologicalcharacteristics of almondrootstocks were studiedin thedepartment ofHorticulture, Ferdowsi University of Mashhad, in 2011-2012. The experiment was carried out with four replications in a completely random blockdesign to study the effects of two concentrations of EM (0 and 1%, three irrigation levels (normal irrigation 100%-control-and irrigation after depletion of 33 and 66% of available water, and four almond rootstocks including GF

  12. Agile Blocker and Clock Jitter Tolerant Low-Power Frequency Selective Receiver with Energy Harvesting Capability.

    Science.gov (United States)

    Hasan, Abul; Helaoui, Mohamed; Ghannouchi, Fadhel M

    2017-08-29

    In this article, a novel tunable, blocker and clock jitter tolerant, low power, quadrature phase shift frequency selective (QPS-FS) receiver with energy harvesting capability is proposed. The receiver's design embraces and integrates (i) the baseband to radio frequency (RF) impedance translation concept to improve selectivity over that of conventional homodyne receiver topologies and (ii) broadband quadrature phase shift circuitry in the RF path to remove an active multi-phase clock generation circuit in passive mixer (PM) receivers. The use of a single local oscillator clock signal with a passive clock division network improves the receiver's robustness against clock jitter and reduces the source clock frequency by a factor of N, compared to PM receivers using N switches (N≥4). As a consequence, the frequency coverage of the QPS-FS receiver is improved by a factor of N, given a clock source of maximum frequency; and, the power consumption of the whole receiver system can eventually be reduced. The tunable QPS-FS receiver separates the wanted RF band signal from the unwanted blockers/interferers. The desired RF signal is frequency down-converted to baseband, while the undesired blocker/interferer signals are reflected by the receiver, collected and could be energy recycled using an auxiliary energy harvesting device.

  13. Selection for Improved Energy Use Efficiency and Drought Tolerance in Canola Results in Distinct Transcriptome and Epigenome Changes.

    Science.gov (United States)

    Verkest, Aurine; Byzova, Marina; Martens, Cindy; Willems, Patrick; Verwulgen, Tom; Slabbinck, Bram; Rombaut, Debbie; Van de Velde, Jan; Vandepoele, Klaas; Standaert, Evi; Peeters, Marrit; Van Lijsebettens, Mieke; Van Breusegem, Frank; De Block, Marc

    2015-08-01

    To increase both the yield potential and stability of crops, integrated breeding strategies are used that have mostly a direct genetic basis, but the utility of epigenetics to improve complex traits is unclear. A better understanding of the status of the epigenome and its contribution to agronomic performance would help in developing approaches to incorporate the epigenetic component of complex traits into breeding programs. Starting from isogenic canola (Brassica napus) lines, epilines were generated by selecting, repeatedly for three generations, for increased energy use efficiency and drought tolerance. These epilines had an enhanced energy use efficiency, drought tolerance, and nitrogen use efficiency. Transcriptome analysis of the epilines and a line selected for its energy use efficiency solely revealed common differentially expressed genes related to the onset of stress tolerance-regulating signaling events. Genes related to responses to salt, osmotic, abscisic acid, and drought treatments were specifically differentially expressed in the drought-tolerant epilines. The status of the epigenome, scored as differential trimethylation of lysine-4 of histone 3, further supported the phenotype by targeting drought-responsive genes and facilitating the transcription of the differentially expressed genes. From these results, we conclude that the canola epigenome can be shaped by selection to increase energy use efficiency and stress tolerance. Hence, these findings warrant the further development of strategies to incorporate epigenetics into breeding. © 2015 American Society of Plant Biologists. All Rights Reserved.

  14. Severe selective magnesium malabsorption: tests of tolerance of oral magnesium supplements.

    Science.gov (United States)

    Mettey, R; Guillard, O; Merle, P; Maillet-Picker, F

    1990-12-01

    Since his birth, we have been monitoring a 12-year-old boy suffering from selective severe magnesium malabsorption. Our essential problem is to prepare a form of galena with acceptable taste, tolerated by the digestive tract and well absorbed; also, the carrier compound must not cause short- or long-term side effects. An additional factor is the steadily increasing need for magnesium from 1 mmol/kg.d at 1 year to 14 mmol/kg.d at present age (345 mg/kg.d). The galena forms currently on sale were, with the exception of lactate and pyrollidone carboxylate, immediately rejected since they contain insufficient Mg2+. Following short trials resulting in diarrhoea, the other two preparations were also rejected. We then constituted - and also abandoned - our own galena compounds: aspartate (bitterness), aspartate + glycerophosphate (GLP) (bitterness), glutamate + GLP ('Chinese restaurant syndrome' and fear of the long term toxic effect of the glutamate), gluconate (excessive volume: 11/1 proportion with Mg2+). A recent test featuring GLP of Mg 40 g + cocoa butter 40 g + cocoa 10 g, brought about vomiting and diarrhoea, and was not adequately absorbed. The best tolerated formula is: Mg GLP 21.33 g; saccharose 6 g; aspartam 1 g; gelatin 0.5 g; citric acid, conserving agent, fruity aroma; water: qs 100 g. Such composition yields a caramel cream absorbed in five small portions, at a daily quantity of 375 g (80 g GLP Mg, 10 g Mg2+). Vitamin B6, which promotes intestinal absorption of magnesium, must be given separately in tablet form at a dose of 1 g/d, since it causes nausea if it is included in the Mg preparation.(ABSTRACT TRUNCATED AT 250 WORDS)

  15. Effects of endotoxin on the lactating mouse

    International Nuclear Information System (INIS)

    Carr, J.K.

    1985-01-01

    The regulation of endogenous mouse mammary tumor virus (MMTV) sequences in trans by a host gene, the Lps locus on mouse chromosome 4, was suspected from a genetic linkage analysis. The Lps locus mediates the mouse's response to the injection of lipopolysaccharide (LPS) in the responder mouse while mice with the deficient allele are incapable of responding. Others have found that endotoxin exposure reduces milk production in lactating animals. This observation was confirmed in mice and extended by examining 125 I-prolactin binding to liver membranes of lactating mice. Endotoxin treatment of responder mice increases liver prolactin binding within 15 minutes, followed by a decline over 6 hours. Scatchard analysis shows that the immediate increase comes from both increased affinity and abundance of the prolactin receptor. No such change in prolactin binding is seen in the non-responder following endotoxin treatment nor in 125 I-insulin binding in responders

  16. The effect of endotoxin on preirradiated mice

    Energy Technology Data Exchange (ETDEWEB)

    Oehlert, W.; Oehlert, M. (Freiburg Univ. (Germany). Inst. fuer Pathologie); Moenig, H.; Konermann, G. (Freiburg Univ. (Germany). Inst. fuer Biophysik und Strahlenbiologie)

    1992-12-01

    Adult male mice were given a whole body irradiation with non-lethal doses of 2.5 or 5 Gy. Unirradiated animals served as controls. The animals (including controls) received a single injection of endotoxin (LPS from Salmonella abortus equi) with doses of 100, 200 or 400 [mu]g one day up to one year after irradiation. Twelve, 24 or 48 hours after lipopolysaccharide (LPS) application the animals were killed and dissected. Animals which died spontaneously were also examined. Liver, lung, kidney, small intestine, and stomach were histologically investigated. The histological findings showed, that differences exist between irradiated and unirradiated mice and that the cause of death is also different for animals dying spontaneously. The investigations have shown that after irradiation phases of different degrees of sensitivity with regard to the endotoxin response exist. This behaviour can be observed by different lethality rates or in the light of the histological results. Moreover, the histological findings have shown, that distinct regenerative changes occur first of all in the liver, in the mucosa of small intestine, and the gastric mucosa, in which the number of differntiated cells compared with the mitotic active cells is reduced. It can be ascertained, that a whole body irradiation with 2.5 to 5 Gy enhances an additional injury by endotoxin weeks to months later. Contrary to this a preirradiation a few days before endotoxin application leads to a 'protection' against the efficacy of endotoxin. These findings can be explained by modes of action described in literature, according to which endotoxins induce the formation of highly active mediators especially the tumor necrosis factor. (orig.).

  17. [Selection and identification of salt tolerant line of sainfoin from the seeds of first post-flight plants].

    Science.gov (United States)

    Xu, Y Y; Wang, M G; Jia, J F

    2001-03-01

    Seeds of sainfoin (Onobrychis vicaefolia Scop.) were carried in the recoverable satellite 940703 and recovered from earth orbit from China in 1994. The progeny seeds were obtained by producing in field. The salt tolerant calluses were selected by screening seedling and callus on 1.5% NaCl-containing medium, reviving growth on NaCl- free medium and selecting callus on 1.2% NaCl-containing medium. The salt tolerant line callus maintained the normal ability to regenerate plant. The salt tolerant line callus exhibited cross-resistance to PEG stress. The variant appeared higher efficiency than control to accumulate proline under salt stress, however, under nonstress condition it had lower proline level than control, which suggested that the higher efficiency to synthesize proline under stress condition may be more important than higher level in tissue under nonstress condition. The mechanism of proline synthesis in the selected callus was considered to result from the alteration in gene sensitivity to water regulation at transcription level. Acrylamide gradient electrophoresis showed that new isoenzyme form with MW 175 kD and 75 kD of superoxide dismutase and esterase respectively appeared in salt torlerant callus. It was indicated that the combination of space mutagenesis with tissue culture could be used for the selection of salt tolerant sainfoin line in vitro.

  18. 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.

  19. Effect of early enteral nutrition (EN) on endotoxin in serum and intestinal permeability in patients with severe acute pancreatitis.

    Science.gov (United States)

    Shen, Q-X; Xu, G-X; Shen, M-H

    2017-06-01

    This work aimed at studying the effect of early enteral nutrition (EN) on serum endotoxin and intestinal permeability in patients with severe acute pancreatitis. 70 cases of patients with severe acute pancreatitis were cured in our hospital from April 2015 to January 2016. Patients selected were randomly divided into two groups including a group of patients having parenteral nutrition (group PN) and that had enteral nutrition (group EN). The results were assessed by: 1) the differences of serum endotoxin level; 2) the differences of the lactulose/mannitol ratio of urine, before intervention and one and two weeks after the intervention. Before the intervention, both groups had similar levels of serum endotoxin and the same lactulose/mannitol excretion rate of urine (p>0.05). One and two weeks after the intervention, the serum endotoxin level and the lactulose/mannitol excretion rate of urine of the group PN were significantly higher than the group EN (pEN has a bigger effect on serum endotoxin and intestinal permeability in patients with severe acute pancreatitis. EN can better promote the elimination of serum endotoxin and reduce intestinal permeability. Therefore, EN deserves clinical expansion.

  20. Selective labelling and eradication of antibiotic-tolerant bacterial populations in Pseudomonas aeruginosa biofilms

    Science.gov (United States)

    Chua, Song Lin; Yam, Joey Kuok Hoong; Hao, Piliang; Adav, Sunil S.; Salido, May Margarette; Liu, Yang; Givskov, Michael; Sze, Siu Kwan; Tolker-Nielsen, Tim; Yang, Liang

    2016-01-01

    Drug resistance and tolerance greatly diminish the therapeutic potential of antibiotics against pathogens. Antibiotic tolerance by bacterial biofilms often leads to persistent infections, but its mechanisms are unclear. Here we use a proteomics approach, pulsed stable isotope labelling with amino acids (pulsed-SILAC), to quantify newly expressed proteins in colistin-tolerant subpopulations of Pseudomonas aeruginosa biofilms (colistin is a ‘last-resort' antibiotic against multidrug-resistant Gram-negative pathogens). Migration is essential for the formation of colistin-tolerant biofilm subpopulations, with colistin-tolerant cells using type IV pili to migrate onto the top of the colistin-killed biofilm. The colistin-tolerant cells employ quorum sensing (QS) to initiate the formation of new colistin-tolerant subpopulations, highlighting multicellular behaviour in antibiotic tolerance development. The macrolide erythromycin, which has been previously shown to inhibit the motility and QS of P. aeruginosa, boosts biofilm eradication by colistin. Our work provides insights on the mechanisms underlying the formation of antibiotic-tolerant populations in bacterial biofilms and indicates research avenues for designing more efficient treatments against biofilm-associated infections. PMID:26892159

  1. Physiological and molecular analysis of selected Kenyan maize lines for aluminum tolerance

    Science.gov (United States)

    Aluminum (Al) toxicity is an important limitation to maize production in many tropical and sub-tropical acid soil areas. The aim of this study was to survey the variation in Al tolerance in a panel of maize lines adapted for Kenya and look for novel sources of Al tolerance. 112 Kenyan maize accessio...

  2. Use of Heat Stress Responsive Gene Expression Levels for Early Selection of Heat Tolerant Cabbage (Brassica oleracea L.

    Directory of Open Access Journals (Sweden)

    Jun Cheul Ahn

    2013-06-01

    Full Text Available Cabbage is a relatively robust vegetable at low temperatures. However, at high temperatures, cabbage has disadvantages, such as reduced disease tolerance and lower yields. Thus, selection of heat-tolerant cabbage is an important goal in cabbage breeding. Easier or faster selection of superior varieties of cabbage, which are tolerant to heat and disease and have improved taste and quality, can be achieved with molecular and biological methods. We compared heat-responsive gene expression between a heat-tolerant cabbage line (HTCL, “HO”, and a heat-sensitive cabbage line (HSCL, “JK”, by Genechip assay. Expression levels of specific heat stress-related genes were increased in response to high-temperature stress, according to Genechip assays. We performed quantitative RT-PCR (qRT-PCR to compare expression levels of these heat stress-related genes in four HTCLs and four HSCLs. Transcript levels for heat shock protein BoHsp70 and transcription factor BoGRAS (SCL13 were more strongly expressed only in all HTCLs compared to all HSCLs, showing much lower level expressions at the young plant stage under heat stress (HS. Thus, we suggest that expression levels of these genes may be early selection markers for HTCLs in cabbage breeding. In addition, several genes that are involved in the secondary metabolite pathway were differentially regulated in HTCL and HSCL exposed to heat stress.

  3. Endotoxin exposure assessment - measurement and characterization

    NARCIS (Netherlands)

    Spaan, S.

    2008-01-01

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

  4. MDP-Induced selective tolerance to TLR4 ligands: impairment in NOD2 mutant Crohn's disease patients.

    Science.gov (United States)

    Cantó, Elisabet; Moga, Esther; Ricart, Elena; Garcia-Bosch, Orlando; Garcia-Planella, Esther; Juarez, Candido; Vidal, Silvia

    2009-11-01

    Pathogen infection is a complex process in which several pathogen-recognition receptor (PRR) pathways are activated to induce proinflammatory mediators. The activation of multiple PRRs suggests an interaction between Toll-like receptors (TLRs) and nucleotide-binding oligomerization domain-like receptor (NOD) signaling pathways. To understand the modulation induced by NOD2 signals on successive responses to pathogen-associated molecular patterns (PAMPs), we examined how muramyl dipeptide (MDP) pretreatment reprograms the MDP+LPS (lipopolysaccharide) response of monocytes from human peripheral blood. Preexposure to bacterial MDP components induced selective tolerance to a subsequent NOD2+TLR4 stimulation. MDP pretreatment inhibited the production of tumor necrosis factor alpha (TNFalpha) and interleuken 10 (IL10), whereas IL6 and IL8 remained unaffected. MDP-induced tolerance was independent of receptor downregulation but was associated with reduced levels of phosphorylated TAK1 and abrogated phosphorylation of the downstream MAPK.Since Nod2 mutations have been associated with susceptibility to develop Crohn's disease (CD), we compared the MDP-induced tolerance in healthy donors and CD patients with compound heterozygous Nod2 mutations (Mut-Nod2) expressing variant NOD2 proteins. MDP-induced tolerance in Mut-Nod2 patients reduced IL10 but not TNFalpha production. In contrast with healthy donors, a p38-independent TNFalpha production was observed during the kinetics of the MDP+LPS response in Mut-Nod2 patients. Our findings suggest that the selective tolerance induced by MDP in healthy donors was related to the modulation of a convergent nub of NOD2 and TLR4 signaling pathways. This MDP-induced tolerance was impaired in Mut-Nod2 CD patients, resulting in a p38-independent TNFalpha production and an imbalance between pro- and antiinflammatory cytokines that could be partly responsible for the pathogenesis of CD.

  5. 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.

  6. Defects-tolerant Co-Cr-Mo dental alloys prepared by selective laser melting.

    Science.gov (United States)

    Qian, B; Saeidi, K; Kvetková, L; Lofaj, F; Xiao, C; Shen, Z

    2015-12-01

    CrCoMo alloy specimens were successfully fabricated using selective laser melting (SLM). The aim of this study was to carefully investigate microstructure of the SLM specimens in order to understand the influence of their structural features inter-grown on different length scales ranging from nano- to macro-levels on their mechanical properties. Two different sets of processing parameters developed for building the inner part (core) and the surface (skin) of dental prostheses were tested. Microstructures were characterized by SEM, EBSD and XRD analysis. The elemental distribution was assessed by EDS line profile analysis under TEM. The mechanical properties of the specimens were measured. The microstructures of both specimens were characterized showing formation of grains comprised of columnar sub-grains with Mo-enrichment at the sub-grain boundaries. Clusters of columnar sub-grains grew coherently along one common crystallographic direction forming much larger single crystal grains which are intercrossing in different directions forming an overall dendrite-like microstructure. Three types of microstructural defects were occasionally observed; small voids (10 μm). Despite the presence of these defects, the yield and the ultimate tensile strength (UTS) were 870 and 430MPa and 1300MPa and 1160MPa, respectively, for the skin and core specimens which are higher than casted dental alloy. Although the formation of microstructural defects is hard to be avoided during the SLM process, the SLM CoCrMo alloys can achieve improved mechanical properties than their casted counterparts, implying they are "defect-tolerant". Copyright © 2015 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

  7. Selection of efficient salt-tolerant bacteria containing ACC deaminase for promotion of tomato growth under salinity stress

    Directory of Open Access Journals (Sweden)

    Kannika Chookietwattana* and Kedsukon Maneewan

    2012-05-01

    Full Text Available For successful application of plant growth promoting bacteria (PGPB in salt-affected soil, bioinoculant with salt-tolerant property is required in order to provide better survival and perform well in the field. The present study aimed to select the most efficient salt-tolerant bacterium containing 1-aminocyclopropane-1-carboxylic acid (ACC deaminase from eighty four bacterial strains and to investigate the effects of the selected bacterium on the germination and growth of tomato (Licopersicon esculentum Mill. cv. Seeda under saline conditions. The Bacillus licheniformis B2r was selected for its ability to utilize ACC as a sole nitrogen source under salinity stress. It also showed a high ACC deaminase activity at 0.6 M NaCl salinity. Tomato plants inoculated with the selected bacterium under various saline conditions (0, 30, 60, 90 and 120 mM NaCl revealed a significant increase in the germination percentage, germination index, root length, and seedling dry weight especially at salinity levels ranging from 30-90 mM NaCl. The work described in this report is an important step in developing an efficient salt-tolerant bioinoculant to facilitate plant growth in saline soil.

  8. Mediated effect of endotoxin and lead upon hepatic metabolism

    International Nuclear Information System (INIS)

    Kuttner, R.E.; Ebata, T.; Schumer, W.

    1984-01-01

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

  9. Selection of high temperature and salinity tolerant Trichoderma isolates with antagonistic activity against Sclerotium rolfsii.

    Science.gov (United States)

    Poosapati, Sowmya; Ravulapalli, Prasad Durga; Tippirishetty, Navaneetha; Vishwanathaswamy, Dinesh Kumar; Chunduri, Sarada

    2014-01-01

    Trichoderma isolates were collected from varied agro-climatic zones of India and screened for high temperature and salinity tolerance. Among all the isolates tested, T. asperellum, TaDOR673 was highly tolerant to heat shock of 52°C with a mean spore count (log c.f.u/ml) of 4.33. The isolate after recovery from heat shock possessed higher germination rate and biomass production compared to its wild counterpart, upon prolonged exposure to 37°C. Under stress, TaDOR673 accumulated >15% of trehalose and >5% of mannose and raffinose compared to the wild type strain signifying their role in stress tolerance. T. asperellum, TaDOR693 and T. asperellum, TaDORS3 were identified as superior salt-tolerant isolates. Interestingly, TaDOR673 also possessed similar tolerance levels to increasing saline concentrations as indicated by its improved colony growth under stress conditions. T. asperellum, TaDOR673 and T. asperellum, TaDOR7316 effectively controlled the collar rot disease in groundnut by 79.7% when screened in vitro and in vivo. Thus, the study identified a potential thermotolerant and saline tolerant strain of Trichoderma, TaDOR673 that could be used as potential bioagent in stressed soils.

  10. Effective selection criteria for screening drought tolerant recombinant inbred lines of sunflower

    Directory of Open Access Journals (Sweden)

    Abdi Nishtman

    2013-01-01

    Full Text Available In this study, seventy two sunflower recombinant inbred lines were tested for their yielding ability under both water-stressed and well-watered states. The inbred lines were evaluated in a rectangular 8´9 lattice design with two replications in both well-watered and water-stressed conditions, separately. Eight drought tolerance indices including stability tolerance index (STI, mean productivity (MP, geometric mean productivity (GMP, harmonic mean (HM, stress susceptibility index (SSI, tolerance index (TOL, yield index (YI and yield stability index (YSI were calculated based on grain yield for every genotype. Results showed the highest values of mean productivity (MP index, geometric mean productivity (GMP, yield index (YI, harmonic mean (HM and stress tolerance index (STI indices for ‘C134a’ inbred line and least values of stress susceptibility index (SSI and tolerance (TOL for C61 inbred line. According to correlation of indices with yield performance under both drought stress and non-stress states and principle component analysis, indices including HM, MP, GMP and STI could properly distinguish drought tolerant sunflower inbred lines with high yield performance under both states. Cluster analysis of inbred lines using Ys, Yp and eight indices, categorized them into four groups including 19, 6, 26 and 19 inbred lines.

  11. Evidence that a herbivore tolerance response affects selection on floral traits and inflorescence architecture in purple loosestrife (Lythrum salicaria).

    Science.gov (United States)

    Thomsen, Christina J M; Sargent, Risa D

    2017-06-01

    The study of the evolution of floral traits has generally focused on pollination as the primary driver of selection. However, herbivores can also impose selection on floral traits through a variety of mechanisms, including florivory and parasitism. Less well understood is whether floral and inflorescence architecture traits that influence a plant's tolerance to herbivory, such as compensatory regrowth, alter pollinator-mediated selection. Because herbivore damage to Lythrum salicaria meristems typically leads to an increase in the number of inflorescences and the size of the floral display, an experiment was conducted to test whether simulated herbivory (i.e. clipping the developing meristem) could alter the magnitude or direction of pollinator-mediated selection on a suite of floral and inflorescence architecture traits. Using a pollen supplementation protocol, pollen limitation was compared in the presence and absence of meristem damage in order to quantify any interaction between pollinator and herbivore-mediated selection on floral traits. Surprisingly, in spite of an obvious impact on floral display and architecture, with clipped plants producing more inflorescences and more flowers, there was no difference in pollen limitation between clipped and unclipped plants. Correspondingly, there was no evidence that imposing herbivore damage altered pollinator-mediated selection in this system. Rather, the herbivory treatment alone was found to alter direct selection on floral display, with clipped plants experiencing greater selection for earlier flowering and weaker selection for number of inflorescences when compared with unclipped plants. These findings imply that herbivory on its own can drive selection on plant floral traits and inflorescence architecture in this species, even more so than pollinators. Specifically, herbivory can impose selection on floral traits if such traits influence a plant's tolerance to herbivory, such as through the timing of flowering

  12. 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, Ayşe Nihal; Malik, Kafait U.; Tunctan, Bahar

    2011-01-01

    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 due to 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 PGI2, PGE2, 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-PGF1α and PGE2 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. PMID:21463481

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

    NARCIS (Netherlands)

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

    2012-01-01

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

  14. Biochemical principle of Limulus test for detecting bacterial endotoxins

    OpenAIRE

    Iwanaga, Sadaaki

    2007-01-01

    A hemocyte lysate from horseshoe crab (Limulus) produced a gel, when exposed to Gram-negative bacterial endotoxins, lipopolysaccharides (LPS). This gelation reaction of the lysate, so-called Limulus test, has been widely employed as a simple and very sensitive assay method for endotoxins. Recent biochemical studies on the principle of Limulus test indicate that the hemocytes contain several serine protease zymogens, which constitute a coagulation cascade triggered by endotoxins, and that ther...

  15. Selection of high temperature and salinity tolerant Trichoderma isolates with antagonistic activity against Sclerotium rolfsii

    OpenAIRE

    Poosapati, Sowmya; Ravulapalli, Prasad Durga; Tippirishetty, Navaneetha; Vishwanathaswamy, Dinesh Kumar; Chunduri, Sarada

    2014-01-01

    Trichoderma isolates were collected from varied agro-climatic zones of India and screened for high temperature and salinity tolerance. Among all the isolates tested, T. asperellum, TaDOR673 was highly tolerant to heat shock of 52°C with a mean spore count (log c.f.u/ml) of 4.33. The isolate after recovery from heat shock possessed higher germination rate and biomass production compared to its wild counterpart, upon prolonged exposure to 37°C. Under stress, TaDOR673 accumulated >15% of trehalo...

  16. EFFECTS ON ADH ACTIVITY AND DISTRIBUTION, FOLLOWING SELECTION FOR TOLERANCE TO ETHANOL IN DROSOPHILA-MELANOGASTER

    NARCIS (Netherlands)

    KERVER, JWM; WOLF, W; KAMPING, A; VANDELDEN, W

    1992-01-01

    Strains of Drosophila melanogaster homozygous for either the Adh(F) or the Adh(S) allele were kept on food supplemented with ethanol for 20 generations. These strains (FE and SE) were tested for tolerance to ethanol and compared with control strains (FN and SN). The E strains showed increased

  17. Soil moisture deficit selects for desiccation tolerant Rhizobium leguminosarum bv. trifolii

    NARCIS (Netherlands)

    Ham, van Robert; O'Callaghan, Maureen; Geurts, Rene; Ridgway, Hayley J.; Ballard, Ross; Noble, Alasdair; Macara, Gregor; Wakelin, Steven A.

    2016-01-01

    Tolerance to desiccation is a highly desirable attribute for Rhizobium spp., which are widely used as symbionts of pasture legumes. Poor survival of Rhizobium spp. is of particular concern during the commercial formulation and seed application phase, but also when sown into dry fields and during

  18. Selection and identification of air pollution-tolerant plants by air ...

    African Journals Online (AJOL)

    The study examined air pollution tolerance index (APTI) of ten tree species in urban parks of Isfahan, Iran. Four physiological and biochemical parameters namely: leaf relative water content of leaf (RWC), ascorbic acid content, total leaf chlorophyll (TCH) and leaf extract pH were used to determined the APTI. The results ...

  19. Genetic variation of Lymnaea stagnalis tolerance to copper: A test of selection hypotheses and its relevance for ecological risk assessment

    International Nuclear Information System (INIS)

    Côte, Jessica; Bouétard, Anthony; Pronost, Yannick; Besnard, Anne-Laure; Coke, Maïra; Piquet, Fabien; Caquet, Thierry; Coutellec, Marie-Agnès

    2015-01-01

    The use of standardized monospecific testing to assess the ecological risk of chemicals implicitly relies on the strong assumption that intraspecific variation in sensitivity is negligible or irrelevant in this context. In this study, we investigated genetic variation in copper sensitivity of the freshwater snail Lymnaea stagnalis, using lineages stemming from eight natural populations or strains found to be genetically differentiated at neutral markers. Copper-induced mortality varied widely among populations, as did the estimated daily death rate and time to 50% mortality (LT50). Population genetic divergence in copper sensitivity was compared to neutral differentiation using the Q ST -F ST approach. No evidence for homogenizing selection could be detected. This result demonstrates that species-level extrapolations from single population studies are highly unreliable. The study provides a simple example of how evolutionary principles could be incorporated into ecotoxicity testing in order to refine ecological risk assessment. - Highlights: • Genetic variation in copper tolerance occurs between Lymnaea stagnalis populations. • We used the Q ST -F ST approach to test evolutionary patterns in copper tolerance. • No evidence for uniform selection was found. • Results suggest that extrapolations to the species level are not safe. • A method is proposed to refine ecological risk assessment using genetic parameters. - Genetic variation in copper tolerance occurs in Lymnaea stagnalis. A method is proposed for considering evolutionary parameters in ecological risk assessment

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

    Science.gov (United States)

    Cooper, James F

    2011-01-01

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

  1. Isolation and cultivation of microalgae select for low growth rate and tolerance to high pH

    DEFF Research Database (Denmark)

    Berge, Terje; Daugbjerg, Niels; Hansen, Per Juel

    2012-01-01

    Harmful microalgal blooms or red tides are often associated with high levels of pH. Similarly, species and strains of microalgae cultivated in the laboratory with enriched media experience recurrent events of high pH between dilutions with fresh medium. To study the potential for laboratory...... selection by high pH, we compared, under identical experimental conditions the upper pH tolerance limits for growth in addition to growth and production rates of 23 strains of the common bloom-forming dinoflagellate Heterocapsa triquetra. The strains had been cultivated in official culture centres from ca....... 1 to 51 years (corresponding to 200–10,000 generations). Strains cultivated for less than 10 years had significantly lower mean and median upper pH tolerance limits for growth, and higher growth and production rates compared to strains cultivated for more than 20 years. The range and variation...

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

    Directory of Open Access Journals (Sweden)

    Enrique Álvarez

    2017-08-01

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

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

    Science.gov (United States)

    Álvarez, Enrique; Toledano, Víctor; Morilla, Fernando; Hernández-Jiménez, Enrique; Cubillos-Zapata, Carolina; Varela-Serrano, Aníbal; Casas-Martín, José; Avendaño-Ortiz, José; Aguirre, Luis A.; Arnalich, Francisco; Maroun-Eid, Charbel; Martín-Quirós, Alejandro; Quintana Díaz, Manuel; López-Collazo, Eduardo

    2017-01-01

    System dynamics is a powerful tool that allows modeling of complex and highly networked systems such as those found in the human immune system. We have developed a model that reproduces how the exposure of human monocytes to lipopolysaccharides (LPSs) induces an inflammatory state characterized by high production of tumor necrosis factor alpha (TNFα), which is rapidly modulated to enter into a tolerant state, known as endotoxin tolerance (ET). The model contains two subsystems with a total of six states, seven flows, two auxiliary variables, and 14 parameters that interact through six differential and nine algebraic equations. The parameters were estimated and optimized to obtain a model that fits the experimental data obtained from human monocytes treated with various LPS doses. In contrast to publications on other animal models, stimulation of human monocytes with super-low-dose LPSs did not alter the response to a second LPSs challenge, neither inducing ET, nor enhancing the inflammatory response. Moreover, the model confirms the low production of TNFα and increased levels of C–C motif ligand 2 when monocytes exhibit a tolerant state similar to that of patients with sepsis. At present, the model can help us better understand the ET response and might offer new insights on sepsis diagnostics and prognosis by examining the monocyte response to endotoxins in patients with sepsis. PMID:28824640

  4. Salt tolerances of some mainland tree species select as through nursery screening.

    Science.gov (United States)

    Miah, Md Abdul Quddus

    2013-09-15

    A study of salt tolerance was carried out on germination, survival and height growth performance of important mesophytic species such as Acacia auriculiformis, Acacia hybrid, Artocarpus heterophyllus, Albizia procera, Albizia lebbeck, Acacia nilotica, Achras sapota, Casuarina equisetifolaia, Emblica officinalis, Leucaena leucocephala, Samania saman, Swetenia macrophylla, Terminalia arjuna, Tamarindus indica, Terminalia bellirica and Thespesia populnea in nursery stage using fresh water and salt (NaCl) solutions of 10, 15 and 20 ppm. Effect of salt on germination, survival performance and height growth performance were examined in this condition. Based on the observation, salt tolerance of these species has been determined Acacia auriculiformis, Acacia hybrid, Achras sapota, Casuarina equisetifolia, Leucaena leucocephala and Tamarindus indica has showed the best capacity to perform in different salinity conditions. Acacia nilotica, Emblica officinalis, Thespesia populnea has performed better. Albizia procera, Samania saman and Terminalia bellirica, germination and height performance showed good but when salinity increases survivability were decreases.

  5. Relative efficacy and tolerability of vortioxetine versus selected antidepressants by indirect comparisons of similar clinical studies.

    Science.gov (United States)

    Llorca, Pierre-Michel; Lançon, Christophe; Brignone, Mélanie; Rive, Benoît; Salah, Samir; Ereshefsky, Larry; Francois, Clément

    2014-12-01

    Vortioxetine is an antidepressant with multimodal activity which has shown efficacy in major depressive disorder (MDD) patients in six of ten short-term, randomized, placebo-controlled trials (completed end 2012). We performed meta-regression analyses to indirectly compare vortioxetine to seven marketed antidepressants with different mechanisms of action. To ensure study comparability, only experimental drug and placebo arms from placebo-controlled registration studies were included in primary analyses. The main outcomes were efficacy (standardized mean difference in change from baseline to 2 months on primary endpoint [MADRS/HAM-D]), and tolerability (withdrawal rate due to adverse events). For efficacy, estimates of treatment effect (negative estimates favor vortioxetine) for vortioxetine versus comparators were: agomelatine, -0.16 (p = 0.11); desvenlafaxine, 0.03 (p = 0.80); duloxetine, 0.09 (p = 0.42); escitalopram, -0.05 (p = 0.70); sertraline, -0.04 (p = 0.83); venlafaxine IR/XR, 0.12 (p = 0.33); and vilazodone, -0.25 (p = 0.11). For tolerability, all but one combination was numerically in favor of vortioxetine (odds ratio antidepressant effect estimates or relative ranking. These meta-regression data show that vortioxetine offers a comparable or favorable combination of efficacy (measured by MADRS/HAM-D) and tolerability (measured by withdrawal rate due to adverse events) versus other antidepressants in registration studies in MDD. Alternative methods like mixed-treatment comparison and inclusion of all randomized studies and active reference arms may provide complementary information to this analysis (more evidence but also more heterogeneity). Key messages: Indirect comparisons based on registration studies allow a useful comparison between a recently approved antidepressant and an approved drug. Vortioxetine offers a comparable or favorable combination of efficacy (measured by MADRS/HAM-D assessments) and tolerability (measured by withdrawal rate due to

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

    DEFF Research Database (Denmark)

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

    1994-01-01

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

  7. 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...

  8. Development of a new heat tolerance index for selecting productive goats for the tropics

    International Nuclear Information System (INIS)

    Kamal, T.H.; Mostafa, S.I.; Habib, A.A.; Elmasry, A.M.; Abdelsamee, A.M.; Abolnaga, A.I.; Kassab, F.A.; Abdelhamid, A.M.

    1988-01-01

    A heat tolerance index previously developed in cattle was verified in two breeds of goats to identify young heat tolerant animals capable of maintaining liveweight and milk yield on exposure to high environmental temperatures. Twelve Baladi and Bedouin goats were divided into two equal groups and offered either river or salt water (1.8% Mediterranean sea salt). The animals were maintained in climatic chambers at 18 deg. C and 70% RH for an initial four day period followed by another four days at 38 deg. C and 50% RH for seven hours per day. On the second day of each period, each animal was injected intravenously with tritiated water and total body water (TBW) determined. The percentage increase in TBW induced by the high temperature treatment was subtracted from 100 and the TBW heat tolerance index (HTI) was calculated. TBW-HTI correlated significantly with the percentage increase in live body weight (LBW) in goats over a three month exposure to heat stress and with the percentage decrease in daily milk yield over a seven day heat exposure period. TBW-HTI averages of Baladi and Bedouin goats drinking river water were 81 and 88 respectively while those drinking salt water were 86 and 92 respectively. The equations for the predicted percentage increase in liveweight (Y) in a hot environment for Baladi and Bedouin goats were Y = -38.56 + 0.728X and Y = -45.27 + 0.622X respectively, where X is the TBW-HTI index; the predicted percentage decrease in milk yield in a hot environment Y for all goats was Y = 142.28-1.339X. (author). 9 refs, 1 fig., 5 tabs

  9. 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.

  10. 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.

  11. Effect of Clonal Selection on Daphnia Tolerance to Dark Experimental Conditions.

    Directory of Open Access Journals (Sweden)

    Sandra J Connelly

    Full Text Available Recent studies have demonstrated substantial effects of environmental stress that vary among clones. Exposure to ultraviolet radiation (UV is an important abiotic stressor that is highly variable in aquatic ecosystems due to diel and seasonal variations in incident sunlight as well as to differences in the UV transparency of water among water bodies, the depth distribution of organisms, and the ability of organisms to detect and respond to UV. In contrast to the convention that all UV is damaging, evidence is accumulating for the beneficial effects of exposure to low levels of UV radiation. Whereas UV has been frequently observed as the primary light-related stressor, herein we present evidence that dark conditions may be similarly "stressful" (reduction of overall fitness, and stress responses vary among clones of the freshwater crustacean Daphnia parvula. We have identified a significant relationship between survivorship and reduced fecundity of clones maintained in dark conditions, but no correlation between tolerance of the clones to dark and UV radiation. Low tolerance to dark conditions can have negative effects not only on accumulated stresses in organisms (e.g. the repair of UV-induced damage in organisms with photolyase, but potentially on the overall physiology and fitness of organisms. Our results support recent evidence of the beneficial effects of low-level UV exposure for some organisms.

  12. Effect of Clonal Selection on Daphnia Tolerance to Dark Experimental Conditions.

    Science.gov (United States)

    Connelly, Sandra J; Stoeckel, James A; Gitzen, Robert A; Williamson, Craig E; González, Maria J

    2016-01-01

    Recent studies have demonstrated substantial effects of environmental stress that vary among clones. Exposure to ultraviolet radiation (UV) is an important abiotic stressor that is highly variable in aquatic ecosystems due to diel and seasonal variations in incident sunlight as well as to differences in the UV transparency of water among water bodies, the depth distribution of organisms, and the ability of organisms to detect and respond to UV. In contrast to the convention that all UV is damaging, evidence is accumulating for the beneficial effects of exposure to low levels of UV radiation. Whereas UV has been frequently observed as the primary light-related stressor, herein we present evidence that dark conditions may be similarly "stressful" (reduction of overall fitness), and stress responses vary among clones of the freshwater crustacean Daphnia parvula. We have identified a significant relationship between survivorship and reduced fecundity of clones maintained in dark conditions, but no correlation between tolerance of the clones to dark and UV radiation. Low tolerance to dark conditions can have negative effects not only on accumulated stresses in organisms (e.g. the repair of UV-induced damage in organisms with photolyase), but potentially on the overall physiology and fitness of organisms. Our results support recent evidence of the beneficial effects of low-level UV exposure for some organisms.

  13. Diallel Analysis and Growth Parameters as Selection Tools for Drought Tolerance in Young Theobroma cacao Plants.

    Science.gov (United States)

    Dos Santos, Emerson Alves; Almeida, Alex-Alan Furtado de; Ahnert, Dario; Branco, Marcia Christina da Silva; Valle, Raúl René; Baligar, Virupax C

    2016-01-01

    This study aimed to estimate the combining ability, of T. cacao genotypes preselected for drought tolerance through diallel crosses. The experiment was conducted under greenhouse conditions at the Cacao Research Center (CEPEC), Ilhéus, Bahia, Brazil, in a completely randomized block design, in an experimental arrangement 21 x 2 [21 complete diallel crosses and two water regimes (control and stressed)]. In the control, soil moisture was kept close to field capacity, with predawn leaf water potential (ΨWL) ranging from -0.1 to -0.5 MPa. In the drought regime, the soil moisture was reduced gradually by decreasing the amount of water application until ΨWL reached -2.0 to -2.5 MPa. Significant differences (p cacao crosses, except for SCA-6 x IMC-67, Catongo x SCA, MOC-01 x Catongo, Catongo x IMC-67 and RB-40 x Catongo. Multivariate analysis showed that stem diameter (CD), total leaf area (TLA), leaf dry biomass (LDB), stem dry biomass (SDB), root dry biomass (RDB), total dry biomass (TDB), root length (RL), root volume (RV), root diameter (RD) cacao genotypes in to tolerant and intolerant to soil water deficit.

  14. Analysis of biological factors for determination of air pollution tolerance index of selected plants in Yamuna Nagar, India.

    Science.gov (United States)

    Sharma, Manju; Panwar, Neeraj; Arora, Pooja; Luhach, Jyoti; Chaudhry, Smita

    2013-05-01

    Air pollution tolerance index (APTI) calculated for various plant species growing in vicinity of three different industrial areas (Paper mill, Sugar mill, Thermal Power Plant) and Yamuna River belt of Yamuna Nagar. Studies were carried out to determine the physiological response of ten plant species. The leaf samples collected from these plant species were used to determine their plant APTI by calculating the ascorbic acid, total chlorophyll, pH, and relative water content for all selected sites. Highest pH, relative water content, ascorbic acid and total chlorophyll was observed in Castor (9.86), Parthenium (96.99%), Ficus benghalensis (14.90 mg g(-1)) and Amaranthus (7.08 mg g(-1)) at Yamuna river, Thermal power plant, Yamuna river and paper mill respectively. It was concluded that out of ten species studied only one species (Ficus benghalensis) showed moderately tolerant response in all selected sites, while other species showed sensitive response. According to observed APTI values, Ficus benghalensis showed the highest value (21.65) at sugar mill followed by thermal power plant (19.38), Paper mill (17.65) and Yamuna River (17.61). The lowest APTI values were reported in Oxalis corniculata (6.42) at Yamuna River belt followed by Malvestrum at sugar mill (7.71).

  15. Constitutive traits and selective indices of Bambara groundnut ( Vigna subterranea (L) Verdc) landraces for drought tolerance under Botswana conditions

    Science.gov (United States)

    Karikari, S. K.; Tabona, T. T.

    Constitutive traits of 12 landraces of Bamabra groundnut ( Vigna subterranea (L) Verdc) were studied during the 1999/2000 and 2000/2001 at the Botswana College of Agriculture, for traits which relate to the crop’s adaptation to drought. The objective was to estimate the degree of phenotypic and genotypic variation of traits whose selection would lead to earliness in maturity and drought tolerance through drought escape. The landraces showed significant diversity. Low environmental variability was recorded for days to maturity, root:shoot ratio, canopy spread, number of seeds per pod, 100-seed weight and shelling percentage. Genotypic correlation coefficients and path coefficient of number of pods per plant and seed size on yield were significantly high. The results of this experiment indicate that, canopy spread, root:shoot ratio, 100-seed weight and number of seeds per pod are among parameters that could be used for indirect selection for drought tolerance in Bambara groundnut in Botswana and other areas where drought is a common occurrence.

  16. 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.

  17. [The protective properties of the endotoxin protein].

    Science.gov (United States)

    Levenson, V I; Belkin, Z P; Egorova, T P

    1991-08-01

    The isolation and properties of endotoxin protein, or lipid A-associated protein (LAP), from Shigella sonnei were described earlier (Zh. mikrobiol. epidemiol. immunobiol., 1991, No. 4, pp. 11-17, and No. 7). In this report the data on its protective activity are presented. In experiments on mice one nanogram of LAP injected i. v. protected 50% of the animals against i. p. challenge with 40 LD50 of virulent S. sonnei. Guinea pigs injected s. c. with 10 micrograms of LAP were protected against local (keratoconjunctival) challenge with S. sonnei, the efficiency of immunization being 58%. LAP preparations containing no detectable amounts of O-antigen (less than 0.003%) were found to have a protective effect. Hyperimmune anti-LAP rabbit serum prevented local infection when incubated with S. sonnei challenge inoculum before injection into guinea pigs. Both active and passive protection induced by LAP was specific since no effect was observed in animals challenged with Shigella flexneri. In the homologous system the protective effect of anti-LAP serum was abolished by the addition of protein-free LPS. These results are compatible with the hypothesis that the protective activity of LAP depends on the presence of minute amounts of O-antigen whose immunogenic effect is greatly amplified by the protein component of the natural endotoxin complex.

  18. Intestinal radiation syndrome: sepsis and endotoxin

    International Nuclear Information System (INIS)

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

    1985-01-01

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

  19. Competitive advantage and tolerance of selected shochu yeast in barley shochu mash.

    Science.gov (United States)

    Takashita, Hideharu; Fujihara, Emi; Furutera, Mihoko; Kajiwara, Yasuhiro; Shimoda, Masahiko; Matsuoka, Masayoshi; Ogawa, Takahira; Kawamoto, Seiji; Ono, Kazuhisa

    2013-07-01

    A shochu yeast strain, Saccharomyces cerevisiae BAW-6, was previously isolated from Kagoshima yeast strain Ko, and has since been utilized in shochu production. The BAW-6 strain carries pho3/pho3 homozygous genes in contrast to the heterozygous PHO3/pho3 genes in the parental Ko strain. However, absence of the PHO3 gene per se cannot explain the fermentation superiority of BAW-6. Here, we demonstrate the growth advantage of the BAW-6 strain over the Ko strain by competitive cultivation in barley shochu preparation, where alcohol yield and nihonshudo of the former strain were higher than those of the latter strain. In addition, the maximum growth rate of BAW-6 was less affected than that of Ko by high Brix values of barley koji medium, suggesting that BAW-6 is less sensitive to growth inhibitory compounds derived from barley or barley koji. The tolerance of BAW-6 to growth inhibitory compounds, cerulenin and diethylstilbestrol (an H⁺-ATPase inhibitor), was also higher than that of other yeast strains. Consistent with BAW-6's tolerance to diethylstilbestrol in the presence of 8% ethanol (pH 4.5), H⁺-ATPase activity, but not transcription of its gene, was higher in BAW-6 than in Ko. We conclude that the BAW-6 strain is associated with certain gene alterations other than PHO3, such that it can maintain cellular ion homeostasis under conditions of ethanol stress during the latter phase of fermentation. Copyright © 2013 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  20. Air pollution tolerance index and heavy metal bioaccumulation in selected plant species from urban biotopes.

    Science.gov (United States)

    Nadgórska-Socha, Aleksandra; Kandziora-Ciupa, Marta; Trzęsicki, Michał; Barczyk, Gabriela

    2017-09-01

    This research was carried out on plants Taraxacum officinale, Plantago lanceolata, Betula pendula and Robinia pseudoacacia growing in urban biotopes with different levels of heavy metal contamination in the city of Dąbrowa Górnicza (southern Poland). Based on the pollution index, the highest heavy metal contamination was determined in the site 4 (connected with industry emitters) and 6 (high traffic). The metal accumulation index (MAI) values ranged within the biotopes in Dąbrowa Górnicza between 7.3 and 20.6 for R. pseudoacacia, 4.71-23.1 for P. lanceolata, 4.68-28.1 for T. officinale and 10.5-27.2 for B. pendula. Increasing tendency in proline content in biotopes connected with high traffic was found in the leaves of investigated plants (except R. pseudoacacia). Similar tendency was observed for ascorbic acid content in the foliage of the plants as well as in T. officinalle in stands connected industrial emission. Non-protein thiols content increased especially in the leaves of R. pseudoacacia in biotopes with high traffic emissions as well as in T. officinale in stands connected with industry. The mean values of APTI (Air Pollution Tolerance Index) within the city of Dąbrowa Górnicza for investigated plants were found in the following ascending order P. lanceolata urban areas with considerable soil and air contamination, especially with heavy metals. The results indicate that species deemed tolerant according to APTI are suitable plants in barriers areas to combat atmospheric pollution. Copyright © 2017 Elsevier Ltd. All rights reserved.

  1. Effects of endotoxin on mammary secretion of lactating cows

    International Nuclear Information System (INIS)

    Lengemann, F.W.; Pitzrick, M.

    1986-01-01

    The objectives were to describe the magnitude and time course of changes in milk pH, Na, K, lactose, and somatic cells and to determine if paracellular pathways were altered after infusion of Escherichia coli endotoxin (serotype 0128:AB12) to produce inflammation in one-half of the udder of the goat. Intramammary infusion of endotoxin increased pH, number of somatic cells, and Na and decreased K and lactose in milk. Sodium and number of somatic cells were increased by as little as .1μg of endotoxin; .25 μg produced changes in most of the other parameters; maximal effect was elicited by 1μg of endotoxin. The gland response peaked from 5 to 7 h after infusion of endotoxin with an increase in milk cellularity as the only significant effect noted in the control gland. Infusion of [ 14 C]lactose into the gland and [/sup 99m/Tc]albumin into the blood demonstrated that large molecules were more able to cross into and out of udder halves after endotoxin treatment. It is suggested that ion interchange rather than bulk flow across paracellular paths is responsible for changes. In addition, endotoxin appeared to reduce lactose secretion and synthesis

  2. Inactivation of Escherichia coli Endotoxin by Soft Hydrothermal Processing▿

    Science.gov (United States)

    Miyamoto, Toru; Okano, Shinya; Kasai, Noriyuki

    2009-01-01

    Bacterial endotoxins, also known as lipopolysaccharides, are a fever-producing by-product of gram-negative bacteria commonly known as pyrogens. It is essential to remove endotoxins from parenteral preparations since they have multiple injurious biological activities. Because of their strong heat resistance (e.g., requiring dry-heat sterilization at 250°C for 30 min) and the formation of various supramolecular aggregates, depyrogenation is more difficult than sterilization. We report here that soft hydrothermal processing, which has many advantages in safety and cost efficiency, is sufficient to assure complete depyrogenation by the inactivation of endotoxins. The endotoxin concentration in a sample was measured by using a chromogenic limulus method with an endotoxin-specific limulus reagent. The endotoxin concentration was calculated from a standard curve obtained using a serial dilution of a standard solution. We show that endotoxins were completely inactivated by soft hydrothermal processing at 130°C for 60 min or at 140°C for 30 min in the presence of a high steam saturation ratio or with a flow system. Moreover, it is easy to remove endotoxins from water by soft hydrothermal processing similarly at 130°C for 60 min or at 140°C for 30 min, without any requirement for ultrafiltration, nonselective adsorption with a hydrophobic adsorbent, or an anion exchanger. These findings indicate that soft hydrothermal processing, applied in the presence of a high steam saturation ratio or with a flow system, can inactivate endotoxins and may be useful for the depyrogenation of parenterals, including end products and medical devices that cannot be exposed to the high temperatures of dry heat treatments. PMID:19502435

  3. [Application of a bacterial endotoxin test for parenteral drugs].

    Science.gov (United States)

    Ogawa, Y

    1994-01-01

    The Limulus test, which has been adopted as the Test for Bacterial Endotoxins in the JP XII, can detect or quantitate endotoxins of Gram-negative bacterial origin using blood corpuscle extracts (Limulus amebocyte lysate, LAL) of horseshoe crabs (Limulus polyphemus, Tachypleus tridentatus, etc.). It may be conducted by the gel-clot or spectrophotometric (turbidimetric and colorimetric) techniques, the former being based on gel formation due to the activation of LAL by endotoxins. The turbidimetric technique is based on the LAL turbidity change during the gel formation and the colorimetric technique on activation of peptide hydrolytic enzymes in LAL. The Limulus test has been unofficially utilized as a simple and highly sensitive method for the determination of endotoxins in parenteral drugs in lieu of the in vivo Pyrogen Test using rabbits. For the Bacterial Endotoxins Test of the JP XII, the gel-clot technique alone was adopted, the technique being only allowed for Injection. Although most parenteral drugs show inhibition or enhancement in practice, this test can be most easily conducted by eliminating interfering effects through dilution of specimens by a factor not exceeding the maximum valid dilution (MVD) with water. Since MVD is dependent on the sensitivity of applied methodology, the turbidimetric and colorimetric techniques, which are more sensitive than the gel-clot technique, have a distinct advantage. The JP, as the leading Pharmacopoeia for the international harmonization of Bacterial Endotoxins Testing, has presented a "Draft towards International Harmonization of Bacterial Endotoxins Test", whose main purpose is the introduction of supplementary turbidimetric and colorimetric techniques. Under these circumstances the following subjects are discussed: (1) the proposal that, with a view towards international harmonization of the technical requirements of Pharmacopoeias, both the turbidimetric and colorimetic techniques should be included together with

  4. Studies To Examine Potential Tolerability Differences between the 5-HT2C Receptor Selective Agonists Lorcaserin and CP-809101.

    Science.gov (United States)

    Higgins, Guy A; Silenieks, Leonardo B; Patrick, Amy; De Lannoy, Ines A M; Fletcher, Paul J; Parker, Linda A; MacLusky, Neil J; Sullivan, Laura C; Chavera, Teresa A; Berg, Kelly A

    2017-05-17

    Lorcaserin (LOR) is a selective 5-HT 2C receptor agonist that has been FDA approved as a treatment for obesity. The most frequently reported side-effects of LOR include nausea and headache, which can be dose limiting. We have previously reported that in the rat, while LOR produced unconditioned signs characteristic of nausea/malaise, the highly selective 5-HT 2C agonist CP-809101 (CP) produced fewer equivalent signs. Because this may indicate a subclass of 5-HT 2C agonists having better tolerability, the present studies were designed to further investigate this apparent difference. In a conditioned gaping model, a rodent test of nausea, LOR produced significantly higher gapes compared to CP consistent with it having higher emetogenic properties. Subsequent studies were designed to identify features of each drug that may account for such differences. In rats trained to discriminate CP-809101 from saline, both CP and LOR produced full generalization suggesting a similar interoceptive cue. In vitro tests of functional selectivity designed to examine signaling pathways activated by both drugs in CHO (Chinese hamster ovary) cells expressing h5-HT 2C receptors failed to identify evidence for biased signaling differences between LOR and CP. Thus, both drugs showed similar profiles across PLC, PLA 2 , and ERK signaling pathways. In studies designed to examine pharmacokinetic differences between LOR and CP, while drug plasma levels correlated with increasing dose, CSF levels did not. CSF levels of LOR increased proportionally with dose; however CSF levels of CP plateaued from 6 to 12 mg/kg. Thus, the apparently improved tolerability of CP likely reflects a limit to CNS levels attained at relatively high doses.

  5. A perfluorinated covalent triazine-based framework for highly selective and water-tolerant CO2 capture

    KAUST Repository

    Zhao, Yunfeng

    2013-01-01

    We designed and synthesized a perfluorinated covalent triazine-based framework (FCTF-1) for selective CO2 capture. The incorporation of fluorine (F) groups played multiple roles in improving the framework\\'s CO 2 adsorption and separation capabilities. Thermodynamically, the strongly polar C-F bonds promoted CO2 adsorption via electrostatic interactions, especially at low pressures. FCTF-1\\'s CO2 uptake was 1.76 mmol g-1 at 273 K and 0.1 bar through equilibrium adsorption, exceeding the CO2 adsorption capacity of any reported porous organic polymers to date. In addition, incorporating F groups produced a significant amount of ultra-micropores (<0.5 nm), which offered not only high gas adsorption potential but also kinetic selectivity for CO2-N 2 separation. In mixed-gas breakthrough experiments, FCTF-1 exhibited an exceptional CO2-N2 selectivity of 77 under kinetic flow conditions, much higher than the selectivity (31) predicted from single-gas equilibrium adsorption data. Moreover, FCTF-1 proved to be tolerant to water and its CO2 capture performance remained excellent when there was moisture in the gas mixture, due to the hydrophobic nature of the C-F bonds. In addition, the moderate adsorbate-adsorbent interaction allowed it to be fully regenerated by pressure swing adsorption processes. These attributes make FCTF-1 a promising sorbent for CO2 capture from flue gas. © 2013 The Royal Society of Chemistry.

  6. Endotoxin, thrombin, and the Limulus amebocyte lysate test.

    Science.gov (United States)

    Yin, E T

    1975-09-01

    The Limulus amebocyte lysate, a proteinaceous composite isolated from the hemolymph cells of the horseshoe crab (Limulus polyphemus) is sensitive to picogram quantities of Gram-negative bacterial lipopolysaccharides. However, a controversy currently exists as to whether the Limulus amebocyte lysate is specifically sensitive to Gram-negative bacterial endotoxins as a result of a recent report that the blood coagulation protease, thrombin, can mimic endotoxins in the Limulus amebocyte lysate test. Experiments including those employing two highly purified fractions isolated from the Limulus lystae have provided us with evidence that thrombin per se is unable to mimic endotoxin.

  7. Noise tolerant selection by gaze-controlled pan and zoom in 3D

    DEFF Research Database (Denmark)

    Hansen, Dan Witzner; Jensen, Henrik Skovsgaard; Hansen, John Paulin

    2008-01-01

    This paper presents StarGazer - a new 3D interface for gaze-based interaction and target selection using continuous pan and zoom. Through StarGazer we address the issues of interacting with graph structured data and applications (i.e. gaze typing systems) using low resolution eye trackers or small......-size displays. We show that it is possible to make robust selection even with a large number of selectable items on the screen and noisy gaze trackers. A test with 48 subjects demonstrated that users who have never tried gaze interaction before could rapidly adapt to the navigation principles of StarGazer. We...... tested three different display sizes (down to PDA-sized displays) and found that large screens are faster to navigate than small displays and that the error rate is higher for the smallest display. Half of the subjects were exposed to severe noise deliberately added on the cursor positions. We found...

  8. Nutrient availability as a mechanism for selection of antibiotic tolerant Pseudomonas aeruginosa within the CF airway.

    Directory of Open Access Journals (Sweden)

    Lucas R Hoffman

    2010-01-01

    Full Text Available Microbes are subjected to selective pressures during chronic infections of host tissues. Pseudomonas aeruginosa isolates with inactivating mutations in the transcriptional regulator LasR are frequently selected within the airways of people with cystic fibrosis (CF, and infection with these isolates has been associated with poorer lung function outcomes. The mechanisms underlying selection for lasR mutation are unknown but have been postulated to involve the abundance of specific nutrients within CF airway secretions. We characterized lasR mutant P. aeruginosa strains and isolates to identify conditions found in CF airways that select for growth of lasR mutants. Relative to wild-type P. aeruginosa, lasR mutants exhibited a dramatic metabolic shift, including decreased oxygen consumption and increased nitrate utilization, that is predicted to confer increased fitness within the nutrient conditions known to occur in CF airways. This metabolic shift exhibited by lasR mutants conferred resistance to two antibiotics used frequently in CF care, tobramycin and ciprofloxacin, even under oxygen-dependent growth conditions, yet selection for these mutants in vitro did not require preceding antibiotic exposure. The selection for loss of LasR function in vivo, and the associated adverse clinical impact, could be due to increased bacterial growth in the oxygen-poor and nitrate-rich CF airway, and from the resulting resistance to therapeutic antibiotics. The metabolic similarities among diverse chronic infection-adapted bacteria suggest a common mode of adaptation and antibiotic resistance during chronic infection that is primarily driven by bacterial metabolic shifts in response to nutrient availability within host tissues.

  9. A facile solvent-free Synthesis Route for the Assembly of Highly CO2 Selective and H2S tolerant NiSIFSIX Metal-Organic Framework

    KAUST Repository

    Eddaoudi, Mohamed

    2015-07-06

    The development of materials for CO2 capture with high selectivity and high tolerance to H2S is of prime importance for various industrially relevant gas streams (e.g. natural gas and biogas upgrading as well as pre-combustion capture). Here, we report the successful fabrication of a MOF with combined exceptional CO2 capture properties and H2S tolerance, namely Ni SIFSIX based-MOF using both solvothermal and solvent-free methodologies.

  10. Selection of lipase producing yeasts for methanol-tolerant biocatalyst as whole cell application for palm-oil transesterification.

    Science.gov (United States)

    Srimhan, Purimprat; Kongnum, Khanitta; Taweerodjanakarn, Siriporn; Hongpattarakere, Tipparat

    2011-03-07

    Methanol-tolerant lipase producing yeast was successfully isolated and selected thorough ecological screening using palm oil-rhodamine B agar as one step-approach. All 49 lipase-producing yeasts exhibited the ability to catalyze esterification reaction of oleic acid and methanol at 3 molar equivalents. However, only 16 isolates catalyzed transesterification reaction of refined palm oil and methanol. Rhodotorula mucilagenosa P11I89 isolated from oil contaminated soil showed the strongest hydrolytic lipase activity of 1.2U/ml against palm oil. The production of oleic methyl ester and fatty acid methyl ester (FAME) of 64.123 and 51.260% was obtained from esterification and transesterification reaction catalyzed by whole cell of R. mucilagenosa P11I89 in the presence of methanol at 3 molar equivalents against the substrates, respectively. FAME content increased dramatically to 83.29% when 6 molar equivalents of methanol were added. Application of the methanol-tolerant-lipase producing yeast as a whole cell biocatalyst was effectively resolved major technical obstacles in term of enzyme stability and high cost of lipase, leading to the feasibility of green biodiesel industrialization. Copyright © 2010 Elsevier Inc. All rights reserved.

  11. Selection for resistance to white pine blister rust affects the abiotic stress tolerances of limber pine

    Science.gov (United States)

    Patrick J. Vogan; Anna W. Schoettle

    2015-01-01

    Limber pine (Pinus flexilis) mortality is increasing across the West as a result of the combined stresses of white pine blister rust (Cronartium ribicola; WPBR), mountain pine beetle (Dendroctonus ponderosae), and dwarf mistletoe (Arceuthobium cyanocarpum) in a changing climate. With the continued spread of WPBR, extensive mortality will continue with strong selection...

  12. Treatment of sepsis by extracorporeal elimination of endotoxin using polymyxin B-immobilized fiber.

    Science.gov (United States)

    Aoki, H; Kodama, M; Tani, T; Hanasawa, K

    1994-04-01

    Despite the use of potent antibiotics and intensive supportive care, mortality remains high among septic shock patients, especially those with endotoxemia. To remove endotoxin directly from the blood, a material consisting of polymyxin B that is immobilized on fibers (PMX-F) and that can selectively detoxify endotoxin was developed. In a preliminary clinical study, 16 patients with septic multiple organ failure were treated with direct hemoperfusion using a PMX-F column. This therapy significantly decreased the endotoxin level from 76 pg/mL to 21 pg/mL after 2 hours of direct hemoperfusion. The hyperdynamic state of the cardiac index, which is a characteristic of endotoxic shock, returned to normal levels after treatment. In septic shock patients with a systolic pressure of less than 100 mm Hg, the systolic arterial pressure increased significantly from the pretreatment level. The alleviation of fever caused by this therapy continued until the day after treatment. Of the 16 patients who underwent this therapy, 9 were alive 2 weeks after this therapy and 7 patients were discharged from the hospital alive. Hemoperfusion with PMX may be an effective treatment for sepsis and septic shock.

  13. Assay of endotoxin by limulus amebocyte lysate.

    Science.gov (United States)

    Ketchum, P A; Novitsky, T J

    2000-01-01

    Horseshoe crabs fight off infectious agents with a complex array of proteins present in amebocytes, the major cell type in their hemolymph. These amebocytes contain both large and small granules (1). When exposed to bacteria or other infectious agents the amebocytes release proteins into their surroundings by exocytosis. The small granules of Limulus amebocytes contain antibacterial proteins, including polyphemusins and the big defensins (2). The large granules contain the Limulus anti-lipopolysaccharide factor (LALF) and the clot-forming group of serine protease zymogens. Exocytosis is initiated by the reaction of amebocytes with lipopolysaccharide (LPS) from Gram-negative bacteria or other microbial components. LPS is also called endotoxin because it is found in the outer membrane of the gram-negative bacterial cell wall. A solid clot forms in response to the lipid A portion of LPS, thereby walling off the infection site or preventing the loss of blood when the animal is damaged physically (3).

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

    Czech Academy of Sciences Publication Activity Database

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

    2018-01-01

    Roč. 63, č. 3 (2018), s. 283-290 ISSN 0015-5632 Institutional support: RVO:67985807 Keywords : endotoxin * hyperbaric condition Subject RIV: EE - Microbiology, Virology Impact factor: 1.521, year: 2016

  15. Biochemical principle of Limulus test for detecting bacterial endotoxins.

    Science.gov (United States)

    Iwanaga, Sadaaki

    2007-05-01

    A hemocyte lysate from horseshoe crab (Limulus) produced a gel, when exposed to Gram-negative bacterial endotoxins, lipopolysaccharides (LPS). This gelation reaction of the lysate, so-called Limulus test, has been widely employed as a simple and very sensitive assay method for endotoxins. Recent biochemical studies on the principle of Limulus test indicate that the hemocytes contain several serine protease zymogens, which constitute a coagulation cascade triggered by endotoxins, and that there is a (1,3)-β-D-glucan-mediated coagulation pathway which also results in the formation of gel. Up to now, six protein components, designated coagulogen, proclotting enzyme, factor B, factor C, and factor G, all of which are closely associated with the endotoxin-mediated coagulation pathway, have been purified and biochemically characterized. The molecular structures of these proteins have also been elucidated. Moreover, the reconstitution experiments using the isolated clotting factors, factor C, factor B, proclotting enzyme and coagulogen in the presence of endotoxin, leads to the formation of coagulin gel. Here, I will focus on the biochemical principle of Limulus test for detecting bacterial endotoxins, and its activation and regulation mechanism on the LPS-mediated coagulation cascade.

  16. Cellular Effects of Endotoxin in Vitro: Mobility of Endotoxin in the Plasma Membrane of Hepatocytes and Neuroblastoma Cells

    Science.gov (United States)

    1985-01-01

    and detoxification of endotoxin. The liver is also the primary site for gluconeogenesis, ketogenesis and ureagenesis and serves a vital role in...supplying the body’s energy needs. While the effect of endotoxin on hepatic gluconeogenesis and ketogenesis has been studied in some detail (For reviews...Jr. Gluconeogenesis, ureagenesis and ketogenesis during sepsis. JPEN 4:277-285, 1980. 2. Berry, L.J. Bacterial toxins. CRC Crit. Rev. Toxicol. 5:239

  17. Route of Antigen Presentation Can Determine the Selection of Foxp3-Dependent or Foxp3-Independent Dominant Immune Tolerance.

    Science.gov (United States)

    Agua-Doce, Ana; Caridade, Marta; Oliveira, Vanessa G; Bergman, Lisa; Lafaille, Maria C; Lafaille, Juan J; Demengeot, Jocelyne; Graca, Luis

    2018-01-01

    It has been shown that dominant tolerance, namely in transplantation, requires Foxp3 + regulatory T cells. Although most tolerance-inducing regimens rely on regulatory T cells, we found that induction of tolerance to proteins in aluminum hydroxide can be achieved in Foxp3-deficient mice using nondepleting anti-CD4 Abs. This type of tolerance is Ag specific, and tolerant mice retain immune competence to respond to unrelated Ags. We demonstrated with chicken OVA-specific TCR-transgenic mice that the same tolerizing protocol (CD4 blockade) and the same target Ag (OVA) achieves Foxp3-dependent transplantation tolerance to OVA-expressing skin grafts, but Foxp3-independent tolerance when the Ag is provided as OVA-aluminum hydroxide. In the latter case, we found that tolerance induction triggered recessive mechanisms leading to elimination of effector cells and, simultaneously, a dominant mechanism associated with the emergence of an anergic and regulatory CTLA-4 + IL-2 low Foxp3 - T cell population, where the tolerance state is IL-10 dependent. Such Foxp3-independent mechanisms can improve the efficacy of tolerance-inducing protocols. Copyright © 2017 by The American Association of Immunologists, Inc.

  18. Genetically engineered Thompson Seedless grapevine plants designed for fungal tolerance: selection and characterization of the best performing individuals in a field trial.

    Science.gov (United States)

    Rubio, Julia; Montes, Christian; Castro, Álvaro; Álvarez, Catalina; Olmedo, Blanca; Muñoz, Marisol; Tapia, Eduardo; Reyes, Fernando; Ortega, Marcelo; Sánchez, Evelyn; Miccono, María; Dalla Costa, Lorenza; Martinelli, Lucia; Malnoy, Mickael; Prieto, Humberto

    2015-02-01

    The fungi Botrytis cinerea and Erysiphe necator are responsible for gray mold and powdery mildew diseases, respectively, which are among the most devastating diseases of grapes. Two endochitinase (ech42 and ech33) genes and one N-acetyl-β-D-hexosaminidase (nag70) gene from biocontrol agents related to Trichoderma spp. were used to develop a set of 103 genetically modified (GM) 'Thompson Seedless' lines (568 plants) that were established in open field in 2004 and evaluated for fungal tolerance starting in 2006. Statistical analyses were carried out considering transgene, explant origin, and plant response to both fungi in the field and in detached leaf assays. The results allowed for the selection of the 19 consistently most tolerant lines through two consecutive years (2007-2008 and 2008-2009 seasons). Plants from these lines were grafted onto the rootstock Harmony and established in the field in 2009 for further characterization. Transgene status was shown in most of these lines by Southern blot, real-time PCR, ELISA, and immunostrips; the most tolerant candidates expressed the ech42-nag70 double gene construct and the ech33 gene from a local Hypocrea virens isolate. B. cinerea growth assays in Petri dishes supplemented with berry juices extracted from the most tolerant individuals of the selected population was inhibited. These results demonstrate that improved fungal tolerance can be attributed to transgene expression and support the iterative molecular and physiological phenotyping in order to define selected individuals from a population of GM grapevines.

  19. Interval training early after heart failure decompensation is safe and improves exercise tolerance and quality of life in selected patients.

    Science.gov (United States)

    Doletsky, Artem; Andreev, Denis; Giverts, Ilya; Svet, Alexey; Brand, Anna; Kuklina, Maria; Sedov, Vsevolod; Dikur, Oxana; Syrkin, Abram; Saner, Hugo

    2018-01-01

    Aims To evaluate safety and efficacy of moderate intensity interval exercise training early after heart failure decompensation on exercise tolerance and health-related quality of life (HRQoL). Methods and results This is a prospective randomized controlled study. We screened 234 consecutive patients admitted with decompensated heart failure; 46 patients (42 men/4 women; 61 ± 12 years of age) were randomized to a moderate intensity aerobic interval training ( n = 24) or to a control group ( n = 22). Patients underwent cardiopulmonary exercise testing, echocardiography and Minnesota Living with Heart Failure questionnaire (MLHFQ) at baseline, after three weeks and after three months. After three weeks, peak-VO 2 increased by 17% in the training group ( p = 0.003) with further increase by 10% after three months ( p training group (from 64.6 ± 15.6 to 30.8 ± 12.9, p training group, but not in controls. Left ventricular ejection fraction was not significantly different between the two groups at baseline and after three months. No serious adverse events related to exercise testing or training were observed. Conclusions Interval exercise training early after an episode of heart failure decompensation is safe and effective in improving exercise tolerance and health-related quality of life in selected patients after achievement of clinical stability. Positive effects remained sustained after three months. Further studies are needed to define role and indications for interval exercise training early after heart failure decompensation.

  20. Evaluation and Selection for Drought Tolerance in Iranian Fenugreek (Trigonella foenum-graecum Landraces at Germination and Seedling Growth Stages

    Directory of Open Access Journals (Sweden)

    D. Sadeghzadeh Ahari

    2016-07-01

    Full Text Available Introduction: Fenugreek (Trigonella foneum-graecum L. has been cultivated in vegetable farms at the most parts of Iran. It is an annual crop belonging to the Leguminosae family. It originated from west Asia and Iran and cultivated at mostly in European, Asian and African countries, presently. With distinguished of feeding and medicinal values, low needs to soil conditions and its width adaptability to cultivation in different regions, the range of fenugreek cultivation areas have been extended from America to India. In most parts of Iran there is limiting possibilities for cultivation of horticultural and agricultural crops for the reason of limiting water harvesting and unsuitable rainfall distributions. There is no doubt that introduction of new crops for such conditions could increase variation of crops production and stability of farming systems. Plants landraces have been created in thousands of cultivation years under different climatologically and local cropping systems. They are evolved by natural and artificial selection under environmental conditions where they were grown and there have accumulative adaptive genes for tolerance to biotic and abiotic stresses and are the most precious materials in starting of breeding programs. Germination phase is the most important period that guaranties the growth and establishments of crops. One of the basic activators of germination starters is water and limiting of it (drought is the most important retardant of seed growth during germination period under field condition. Fast germination and emergence of seedling from soil and high preliminary growth rate has been known for one of the drought escape mechanisms for most crops such as chickpea, lentil and bean. In breeding programs of crops, using in vitro method is one of the most used methods in germplasm selection for drought tolerance. This study carried out under laboratory condition in order to evaluate some Iranian fenugreek landraces

  1. Endotoxin increases pulmonary vascular protein permeability in the dog

    International Nuclear Information System (INIS)

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

    1986-01-01

    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

  2. Histological analysis of the association between formocresol and endotoxin in the subcutaneous tissue of mice.

    Science.gov (United States)

    Sant'anna, Ana Teresa; Spolidório, Luis Carlos; Ramalho, Lizeti Toledo Oliveira

    2008-01-01

    This study performed a histological analysis of the effect of formocresol associated to endotoxin (LPS) in the subcutaneous connective tissue of mice. Ninety mice were randomly assigned to 3 groups (n=30). Each animal received one plastic tube implant containing endotoxin solution (10 mg/mL), formocresol (original formula) or a mixture of endotoxin and formocresol. The endotoxin and formocresol groups served as controls. The periods of analysis were 7, 15 and 30 days. At each experimental period, tissue samples were collected and submitted to routine processing for histological analysis. Endotoxin and formocresol produced necrosis and chronic inflammation at 7 and 15 days. At 30 days, the endotoxin group showed no necrosis, while in the formocresol group necrosis persisted. The formocresol-endotoxin association produced necrosis and chronic inflammation in the same way as observed with formocresol at all experimental periods. In conclusion, formocresol seems not to be able to inactive the toxic effects of endotoxin in connective tissues.

  3. Modeling Tolerance Development for the Effect on Heart Rate of the Selective S1P1 Receptor Modulator Ponesimod.

    Science.gov (United States)

    Lott, Dominik; Lehr, Thorsten; Dingemanse, Jasper; Krause, Andreas

    2017-09-15

    Ponesimod is a selective sphingosine-1-phosphate-1 (S1P 1 ) receptor modulator currently under investigation for the treatment of multiple sclerosis. S1P receptor modulators reduce heart rate following treatment initiation. This effect disappears with repeated dosing, enabling development of innovative uptitration regimens to optimize patient safety. There are currently no published pharmacokinetic/pharmacodynamic models describing the heart rate reduction of S1P receptor modulators in humans. The model developed here provides quantification of this effect for ponesimod. A direct-effect I max model with estimated maximum reduction of 45%, tolerance development, and circadian variation best described this effect. The pooled data from nine clinical studies enabled characterization of interindividual variability. The model was used to simulate different treatment regimens to compare the effect of high initial doses vs. gradual uptitration with respect to the occurrence of bradycardia. The results indicate a better safety profile when using gradual uptitration. The model allows studying dosing regimens not clinically tested in silico. © 2017 American Society for Clinical Pharmacology and Therapeutics.

  4. Histological analysis of the association between formocresol and endotoxin in the subcutaneous tissue of mice

    OpenAIRE

    Sant'anna, Ana Teresa; Spolidório, Luis Carlos; Ramalho, Lizeti Toledo Oliveira

    2008-01-01

    This study performed a histological analysis of the effect of formocresol associated to endotoxin (LPS) in the subcutaneous connective tissue of mice. Ninety mice were randomly assigned to 3 groups (n=30). Each animal received one plastic tube implant containing endotoxin solution (10 mg/mL), formocresol (original formula) or a mixture of endotoxin and formocresol. The endotoxin and formocresol groups served as controls. The periods of analysis were 7, 15 and 30 days. At each experimental per...

  5. Bacterial endotoxin adhesion to different types of orthodontic adhesives

    Directory of Open Access Journals (Sweden)

    Priscilla Coutinho ROMUALDO

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

  6. A novel match-line selective charging scheme for high-speed, low-power and noise-tolerant content-addressable memory

    KAUST Repository

    Hasan, Muhammad Mubashwar

    2010-06-01

    Content-addressable memory (CAM) is an essential component for high-speed lookup intensive applications. This paper presents a match-line selective charging technique to increase speed and reduce the energy per bit per search while increasing the noise-tolerance. Simulation in TSMC 0.18 μm technology with 64×72 Ternary CAM shows the match-line energy reduction of 45% compared to the conventional currentsaving scheme with the reduction of minimum cycle time by 68% and the improvement of noise-tolerance by 96%.

  7. 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

  8. Influence of different cleaning practices on endotoxin exposure at sewage treatment plants

    NARCIS (Netherlands)

    Visser, M. J. [=Maaike J.; Spaan, S.; Arts, H. J. J. M.; Smit, L. A. M.; Heederik, D. J. J.

    2006-01-01

    Exposure to endotoxin at sewage treatment plants is associated with an increased prevalence of work-related symptoms in sewage workers. Since cleaning activities are regarded as an important determinant of endotoxin exposure, workers' endotoxin exposure levels during different cleaning activities

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

    Lifescience Database Archive (English)

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

  10. ASTHMA AND RHINITIS INDUCED BY SELECTIVE IMMEDIATE REACTIONS TO PARACETAMOL AND NON-STEROIDAL ANTI-INFLAMMATORY DRUGS IN ASPIRIN TOLERANT SUBJECTS

    Directory of Open Access Journals (Sweden)

    Diana Pérez-Alzate

    2016-07-01

    Full Text Available In subjects with non-steroidal anti-inflammatory drugs (NSAIDs- exacerbated respiratory disease (NERD symptoms are triggered by acetyl salicylic acid (ASA and other strong COX-1 inhibitors, and in some cases by weak COX-1 or by selective COX-2 inhibitors. The mechanism involved is related to prostaglandin pathway inhibition and leukotriene release. Subjects who react to a single NSAID and tolerate others are considered selective responders, and often present urticaria and/or angioedema and anaphylaxis (SNIUAA. An immunological mechanism is implicated in these reactions. However, anecdotal evidence suggests that selective responders who present respiratory airway symptoms may also exist.Our objective was to determine if subjects might develop selective responses to NSAIDs/paracetamol that manifest as upper/lower airways respiratory symptoms. For this purpose we studied patients reporting asthma and/or rhinitis induced by paracetamol or a single NSAID that tolerated ASA. An allergological evaluation plus controlled challenge with ASA was carried out. If ASA tolerance was found, we proceeded with an oral challenge with the culprit drug. The appearance of symptoms was monitored by a clinical questionnaire and by measuring FEV1 and/or nasal airways volume changes pre and post challenge. From a total of 21 initial cases, we confirmed the appearance of nasal and/or bronchial manifestations in ten, characterised by a significant decrease in FEV1% and/or a decrease in nasal volume cavity after drug administration. All cases tolerated ASA.This shows that ASA tolerant subjects with asthma and/or rhinitis induced by paracetamol or a single NSAID without skin/systemic manifestations exist. Whether these patients represent a new clinical phenotype to be included within the current classification of hypersensitivity reactions to NSAIDs requires further investigation.

  11. Identification of X-linked quantitative trait loci affecting cold tolerance in Drosophila melanogaster and fine mapping by selective sweep analysis.

    Science.gov (United States)

    Svetec, Nicolas; Werzner, Annegret; Wilches, Ricardo; Pavlidis, Pavlos; Alvarez-Castro, José M; Broman, Karl W; Metzler, Dirk; Stephan, Wolfgang

    2011-02-01

    Drosophila melanogaster is a cosmopolitan species that colonizes a great variety of environments. One trait that shows abundant evidence for naturally segregating genetic variance in different populations of D. melanogaster is cold tolerance. Previous work has found quantitative trait loci (QTL) exclusively on the second and the third chromosomes. To gain insight into the genetic architecture of cold tolerance on the X chromosome and to compare the results with our analyses of selective sweeps, a mapping population was derived from a cross between substitution lines that solely differed in the origin of their X chromosome: one originates from a European inbred line and the other one from an African inbred line. We found a total of six QTL for cold tolerance factors on the X chromosome of D. melanogaster. Although the composite interval mapping revealed slightly different QTL profiles between sexes, a coherent model suggests that most QTL overlapped between sexes, and each explained around 5-14% of the genetic variance (which may be slightly overestimated). The allelic effects were largely additive, but we also detected two significant interactions. Taken together, this provides evidence for multiple QTL that are spread along the entire X chromosome and whose effects range from low to intermediate. One detected transgressive QTL influences cold tolerance in different ways for the two sexes. While females benefit from the European allele increasing their cold tolerance, males tend to do better with the African allele. Finally, using selective sweep mapping, the candidate gene CG16700 for cold tolerance colocalizing with a QTL was identified. © 2010 Blackwell Publishing Ltd.

  12. 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. Copyright © 2015. Published by Elsevier B.V.

  13. Immunomodulatory effects of honey cannot be distinguished from endotoxin

    DEFF Research Database (Denmark)

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

    2008-01-01

    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....

  14. Screening and selection of most potent diazotrophic cyanobacterial isolate exhibiting natural tolerance to rice field herbicides for exploitation as biofertilizer.

    Science.gov (United States)

    Singh, Surendra; Datta, Pallavi

    2006-01-01

    Periodic applications of heavy dosages of herbicides in modern rice-agriculture are a necessary evil for obtaining high crop productivity. Such herbicides are not only detrimental to weeds but biofertilizer strains of diazotrophic cyanobacteria also. It is therefore, essential to screen and select such biofertilizer strains of diazotrophic cyanobacteria exhibiting natural tolerance to common rice-field herbicides that can be further improved by mutational techniques to make biofertilizer technology a viable one. Therefore, efforts have been made to screen five dominant diazotrophic cyanobacterial forms e.g. filamentous heterocystous Nostoc punctiforme , Nostoc calcicola , Anabaena variabilis and unicellular Gloeocapsa sp. and Aphanocapsa sp. along with standard laboratory strain Nostoc muscorum ISU against increasing concentrations (0-100 mg l(-1) of four commercial grade common rice-field herbicides i.e. Arozin, Butachlor, Alachlor and 2,4-D under diazotrophic growth conditions. The lethal and IGC(50) concentrations for all four herbicides tested were found highest for A. variabilis as compared to other test cyanobacteria. The lowest reduction in chlorophyll a content, photosynthetic oxygen evolution, and N(2)-fixation was found in A. variabilis as compared to other rice field isolates and standard laboratory strain N. muscorum ISU. On the basis of prolong survival potential and lowest reductions in vital metabolic activities tested at IGC(50) concentration of four herbicides, it is concluded that A. variabilis is the most potent and promising cyanobacterial isolate as compared with other forms. This could be further improved by mutational techniques for exploitation as most potential and viable biofertilizer strain.

  15. 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.

  16. Establishment of tolerance to commensal bacteria requires a complex microbiota and is accompanied by decreased intestinal chemokine expression

    DEFF Research Database (Denmark)

    Fink, Lisbeth Nielsen; Metzdorff, S. B.; Zeuthen, Louise

    2012-01-01

    Intricate regulation of tolerance to the intestinal commensal microbiota acquired at birth is critical. We hypothesized that epithelial cell tolerance toward early gram-positive and gram-negative colonizing bacteria is established immediately after birth, as has previously been shown for endotoxin...

  17. Dust, endotoxin, fungi, and bacteria exposure as determined by work task, season, and type of plant in a flower greenhouse.

    Science.gov (United States)

    Thilsing, Trine; Madsen, Anne Mette; Basinas, Ioannis; Schlünssen, Vivi; Tendal, Kira; Bælum, Jesper

    2015-03-01

    working with subtasks in the research and development area for Campanula increased bacteria exposure. This study revealed moderate dust exposure levels compared to the levels observed in other greenhouse productions and other occupations with organic dust exposure such as farming. However, high exposures to bacteria and fungi were detected during selected tasks and the proposed health-based endotoxin exposure limit of 90 EU m(-3) was exceeded in 30% of the samples, which may have health implications for the employees. Exposure levels were found to vary depending on the tasks performed, and thereby results can be used to direct task-based initiatives to reduce workplace exposures. © The Author 2014. Published by Oxford University Press on behalf of the British Occupational Hygiene Society.

  18. Binding of /sup 125/I-labeled endotoxin to bovine, canine, and equine platelets and endotoxin-induced agglutination of canine platelets

    Energy Technology Data Exchange (ETDEWEB)

    Meyers, K.M.; Boehme, M.; Inbar, O.

    1982-10-01

    Endotoxin from Escherichia coli O127:B8, Salmonella abortus-equi and S minnesota induced clumping of some canine platelets (PLT) at a final endotoxin concentration of 1 microgram/ml. Endotoxin-induced clumping of canine PLT was independent of PLT energy-requiring processes, because clumping was observed with canine PLT incubated with 2-deoxy-D-glucose and antimycin A. The PLT responded to adenosine diphosphate before, but not after, incubation with the metabolic inhibitors. Endotoxin induced a slight and inconsistant clumping of bovine and equine PLT at high (mg/ml) endotoxin concentration. High-affinity binding sites could not be demonstrated on canine, bovine, and equine PLT, using /sup 125/I-labeled E coli O127:B8 endotoxin. Nonspecific binding was observed and appeared to be due primarily to an extraneous coat on the PLT surface that was removed by gel filtration. The endotoxin that was bound to PLT did not appear to modify PLT function. An attempt to identify plasma proteins that bound physiologically relevant amounts of endotoxin was not successful. The significance of the endotoxin-induced clumping or lack of it on the pathophysiology of endotoxemia is discussed.

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

    OpenAIRE

    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...

  20. 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

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

    Science.gov (United States)

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

    2017-05-01

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

  2. 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

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

    African Journals Online (AJOL)

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

  4. [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.

  5. Tolerating Zero Tolerance?

    Science.gov (United States)

    Moore, Brian N.

    2010-01-01

    The concept of zero tolerance dates back to the mid-1990s when New Jersey was creating laws to address nuisance crimes in communities. The main goal of these neighborhood crime policies was to have zero tolerance for petty crime such as graffiti or littering so as to keep more serious crimes from occurring. Next came the war on drugs. In federal…

  6. Development of new Dutch Elm disease-tolerant selections for restoration of the American Elm in urban and forested landscapes

    Science.gov (United States)

    C.C. Pinchot; C.E. Flower; K.S. Knight; C. Marks; R. Minocha; D. Lesser; K. Woeste; P.G. Schaberg; B. Baldwin; D.M. Delatte; T.D. Fox; N. Hayes-Plazolles; B. Held; K. Lehtoma; S. Long; S. Mattix; A. Sipes; J.M. Slavicek

    2017-01-01

    The goal of our research and development efforts is to restore American elm (Ulmus americana) as a species in both natural and urban landscapes. Accomplishing this goal requires identification/generation of additional American elm cultivars that are tolerant to Dutch elm disease (DED) caused by Ophiostoma ulmi and O. novo-ulmi, and development of methods to reintroduce...

  7. The selection of radiation tolerant electrical/electronic components for gamma radiation environments in the nuclear power industry

    International Nuclear Information System (INIS)

    Garlick, D.R.

    1984-09-01

    This report briefly describes the mechanisms, units and effects of 1 MeV range gamma radiation on electrical/electronic components and materials. Information is tabulated on the gamma radiation tolerance of a wide range of components and materials. A radiation testing service, based at Harwell, is described. Lists of interested manufacturers and organisations are given. (author)

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

    Science.gov (United States)

    Dungan, Robert S; Leytem, April B

    2009-01-01

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

  9. Histopathological and ultrastructural effects of delta-endotoxins of Bacillus thuringiensis serovar israelensis in the midgut of Simulium pertinax larvae (Diptera, Simuliidae

    Directory of Open Access Journals (Sweden)

    CFG Cavados

    2004-08-01

    Full Text Available The bacterium Bacillus thuringiensis (Bt produces parasporal crystals containing delta-endotoxins responsible for selective insecticidal activity on larvae. Upon ingestion, these crystals are solubilized in the midgut lumen and converted into active toxins that bind to receptors present on the microvilli causing serious damage to the epithelial columnar cells. We investigated the effect of these endotoxins on larvae of the Simulium pertinax, a common black fly in Brazil, using several concentrations during 4 h of the serovar israelensis strain IPS-82 (LFB-FIOCRUZ 584, serotype H-14 type strain of the Institute Pasteur, Paris. Light and electron microscope observations revealed, by time and endotoxin concentration, increasing damages of the larvae midgut epithelium. The most characteristic effects were midgut columnar cell vacuolization, microvilli damages, epithelium cell contents passing into the midgut lumen and finally the cell death. This article is the first report of the histopathological effects of the Bti endotoxins in the midgut of S. pertinax larvae and the data obtained may contribute to a better understanding of the mode of action of this bacterial strain used as bioinsecticide against black fly larvae.

  10. Endotoxin Contamination in Nanomaterials Leads to the Misinterpretation of Immunosafety Results

    Directory of Open Access Journals (Sweden)

    Yang Li

    2017-05-01

    Full Text Available Given the presence of engineered nanomaterials in consumers’ products and their application in nanomedicine, nanosafety assessment is becoming increasingly important. In particular, immunosafety aspects are being actively investigated. In nanomaterial immunosafety testing strategies, it is important to consider that nanomaterials and nanoparticles are very easy to become contaminated with endotoxin, which is a widespread contaminant coming from the Gram-negative bacterial cell membrane. Because of the potent inflammatory activity of endotoxin, contaminated nanomaterials can show inflammatory/toxic effects due to endotoxin, which may mask or misidentify the real biological effects (or lack thereof of nanomaterials. Therefore, before running immunosafety assays, either in vitro or in vivo, the presence of endotoxin in nanomaterials must be evaluated. This calls for using appropriate assays with proper controls, because many nanomaterials interfere at various levels with the commercially available endotoxin detection methods. This also underlines the need to develop robust and bespoke strategies for endotoxin evaluation in nanomaterials.

  11. Physiological and biochemical constituents as predictive appreciation for selection of drought tolerant cultivars in wheat (triticum aestivum L.)

    International Nuclear Information System (INIS)

    Jalal-ud-Din; Khan, S.U.; Gurmani, A.R.

    2012-01-01

    A pot study was undertaken to assess the effect of drought stress imposed at various growth stages on growth, physiological and biochemical attributes of wheat. Five commercial wheat cultivars viz. Chakwal-97, Inqalab-91, Margalla-99, NR-234 and Wafaq-2001 were grown in pots. The plants were subjected to three consecutive drought cycles at tillering, pre-anthesis and milky growth stages. Measurements pertaining to various physiological and biochemical parameters such as relative water content (RWC), proline, superoxide dismutase (SOD), membrane stability index (MSI), yield and yield components were made. Significant reduction in grain yield was observed in all the test varieties when drought was imposed at any growth stage. The reduction was highest (39-64%) when stress was imposed at pre-anthesis followed by tillering growth stage. The wheat variety Wafaq-2001 and Inqalab-91 performed better by giving higher yield and produced greater numbers of filled seeds per spike compared to other varieties. Under water stress proline contents were higher in the drought-tolerant cultivar Wafaq-2001. The same variety showed higher membrane stability index and antioxidant enzymes (SOD) activity under drought stress conditions. The results suggest that pre-anthesis growth stage is the most sensitive towards drought stress. Wheat cultivars: Wafaq-2001 and Inqalab-91 showed best tolerance response against drought stress. Higher proline, RWC and SOD activity under drought stress seems to be the most reliable parameters enabling the discrimination of varieties for drought tolerance. (author)

  12. Endotoxin removal by radio frequency gas plasma (glow discharge)

    Science.gov (United States)

    Poon, Angela

    2011-12-01

    Contaminants remaining on implantable medical devices, even following sterilization, include dangerous fever-causing residues of the outer lipopolysaccharide-rich membranes of Gram-negative bacteria such as the common gut microorganism E. coli. The conventional method for endotoxin removal is by Food & Drug Administration (FDA)-recommended dry-heat depyrogenation at 250°C for at least 45 minutes, an excessively time-consuming high-temperature technique not suitable for low-melting or heat-distortable biomaterials. This investigation evaluated the mechanism by which E. coli endotoxin contamination can be eliminated from surfaces during ambient temperature single 3-minute to cumulative 15-minute exposures to radio-frequency glow discharge (RFGD)-generated residual room air plasmas activated at 0.1-0.2 torr in a 35MHz electrodeless chamber. The main analytical technique for retained pyrogenic bio-activity was the Kinetic Chromogenic Limulus Amebocyte Lysate (LAL) Assay, sufficiently sensitive to document compliance with FDA-required Endotoxin Unit (EU) titers less than 20 EU per medical device by optical detection of enzymatic color development corresponding to < 0.5 EU/ml in sterile water extracts of each device. The main analytical technique for identification of chemical compositions, amounts, and changes during sequential reference Endotoxin additions and subsequent RFGD-treatment removals from infrared (IR)-transparent germanium (Ge) prisms was Multiple Attenuated Internal Reflection (MAIR) infrared spectroscopy sensitive to even monolayer amounts of retained bio-contaminant. KimaxRTM 60 mm x 15 mm and 50mm x 15mm laboratory glass dishes and germanium internal reflection prisms were inoculated with E. coli bacterial endotoxin water suspensions at increments of 0.005, 0.05, 0.5, and 5 EU, and characterized by MAIR-IR spectroscopy of the dried residues on the Ge prisms and LAL Assay of sterile water extracts from both glass and Ge specimens. The Ge prism MAIR

  13. Comparison of Endotoxin Exposure Assessment by Bioaerosol Impinger and Filter-Sampling Methods

    OpenAIRE

    Duchaine, Caroline; Thorne, Peter S.; Mériaux, Anne; Grimard, Yan; Whitten, Paul; Cormier, Yvon

    2001-01-01

    Environmental assessment data collected in two prior occupational hygiene studies of swine barns and sawmills allowed the comparison of concurrent, triplicate, side-by-side endotoxin measurements using air sampling filters and bioaerosol impingers. Endotoxin concentrations in impinger solutions and filter eluates were assayed using the Limulus amebocyte lysate assay. In sawmills, impinger sampling yielded significantly higher endotoxin concentration measurements and lower variances than filte...

  14. Endotoxins in indoor air and settled dust in primary schools in a subtropical climate.

    Science.gov (United States)

    Salonen, Heidi; Duchaine, Caroline; Létourneau, Valérie; Mazaheri, Mandana; Clifford, Sam; Morawska, Lidia

    2013-09-03

    Endotoxins can significantly affect the air quality in school environments. However, there is currently no reliable method for the measurement of endotoxins, and there is a lack of reference values for endotoxin concentrations to aid in the interpretation of measurement results in school settings. We benchmarked the "baseline" range of endotoxin concentration in indoor air, together with endotoxin load in floor dust, and evaluated the correlation between endotoxin levels in indoor air and settled dust, as well as the effects of temperature and humidity on these levels in subtropical school settings. Bayesian hierarchical modeling indicated that the concentration in indoor air and the load in floor dust were generally (school environment and the need for further investigation. Metaregression indicated no relationship between endotoxin concentration and load, which points to the necessity for measuring endotoxin levels in both the air and settled dust. Temperature increases were associated with lower concentrations in indoor air and higher loads in floor dust. Higher levels of humidity may be associated with lower airborne endotoxin concentrations.

  15. Respiratory health effects of exposure to low levels of airborne endotoxin - a systematic review.

    Science.gov (United States)

    Farokhi, Azadèh; Heederik, Dick; Smit, Lidwien A M

    2018-02-08

    Elevated endotoxin levels have been measured in ambient air around livestock farms, which is a cause of concern for neighbouring residents. There is clear evidence that occupational exposure to high concentrations of airborne endotoxin causes respiratory inflammation, respiratory symptoms and lung function decline. However, health effects of exposure to low levels of endotoxin are less well described. The aim of this systematic review is to summarize published associations between exposure to relatively low levels of airborne endotoxin and respiratory health endpoints. Studies investigating respiratory effects of measured or modelled exposure to low levels of airborne endotoxin (average respiratory symptoms and lung function. However, considerable heterogeneity existed in the outcomes of the included studies and no overall estimate could be provided by meta-analysis to quantify the possible relationship. Instead, a best evidence synthesis was performed among studies examining the exposure-response relationship between endotoxin and respiratory outcomes. Significant exposure-response relationships between endotoxin and symptoms and FEV 1 were shown in several studies, with no conflicting findings in the studies included in the best evidence synthesis. Significantly different effects of endotoxin exposure were also seen in vulnerable subgroups (atopics and patients with broncho-obstructive disease) and smokers. Respiratory health effects of exposure to low levels of airborne endotoxin (respiratory health effects, especially in vulnerable subgroups of the population.

  16. Endotoxin level measurement in hemodialysis biofilm using "the whole blood assay".

    Science.gov (United States)

    Marion-Ferey, Karine; Leid, Jeff G; Bouvier, Ghislaine; Pasmore, Mark; Husson, Gilles; Vilagines, Rolland

    2005-06-01

    Biofilms have been found on the inner surface of silicone tubing inside dialysis machines. Endotoxin releasing from those biofilms increases the bioincompatibility of dialysis liquids and leads to long-term inflammatory complications among dialysis patients. Endotoxin measurement is recommended for the control of dialysis liquids. This article describes the use of a new method, the Whole Blood Assay (WBA), for endotoxin quantification in dialysis biofilms. Biofilms were suspended in sterile water by scraping the tubing samples. Diluted blood samples from healthy donors were stimulated overnight with the contaminated suspension. Stimulated mononuclear cells released IL-1beta in response to endotoxins. IL-1beta level was then measured using an ultrasensitive ELISA method. We demonstrated a semilogarithmic model in which the optical densities measured after the ELISA assay increases linearly with the levels of endotoxin. This model allowed the determination of the amount of endotoxins in biofilm samples with a detection limit of 0.032 EU/mL. Most of the time, the amounts of endotoxin measured by the WBA were higher than those measured by the Limulus Amoebocyte Lysate (LAL) assay. This study suggested the presence of "endotoxin-like" compounds different from the lipopolysaccharides that are not detected by the LAL assay. We concluded that the LAL is necessary but insufficient to have a representative quantification of endotoxins that could be hazardous to patient health.

  17. Resistance to Antibiotics, Biocides, Preservatives and Metals in Bacteria Isolated from Seafoods: Co-Selection of Strains Resistant or Tolerant to Different Classes of Compounds

    Directory of Open Access Journals (Sweden)

    José L. Romero

    2017-08-01

    Full Text Available Multi-drug resistant bacteria (particularly those producing extended-spectrum β-lactamases have become a major health concern. The continued exposure to antibiotics, biocides, chemical preservatives, and metals in different settings such as the food chain or in the environment may result in development of multiple resistance or co-resistance. The aim of the present study was to determine multiple resistances (biocides, antibiotics, chemical preservatives, phenolic compounds, and metals in bacterial isolates from seafoods. A 75.86% of the 87 isolates studied were resistant to at least one antibiotic or one biocide, and 6.90% were multiply resistant to at least three biocides and at least three antibiotics. Significant (P < 0.05 moderate or strong positive correlations were detected between tolerances to biocides, between antibiotics, and between antibiotics with biocides and other antimicrobials. A sub-set of 30 isolates selected according to antimicrobial resistance profile and food type were identified by 16S rDNA sequencing and tested for copper and zinc tolerance. Then, the genetic determinants for biocide and metal tolerance and antibiotic resistance were investigated. The selected isolates were identified as Pseudomonas (63.33%, Acinetobacter (13.33%, Aeromonas (13.33%, Shewanella, Proteus and Listeria (one isolate each. Antibiotic resistance determinants detected included sul1 (43.33% of tested isolates, sul2 (6.66%, blaTEM (16.66%, blaCTX−M (16.66%, blaPSE (10.00%, blaIMP (3.33%, blaNDM−1 (3.33%, floR (16.66%, aadA1 (20.0%, and aac(6′-Ib (16.66%. The only biocide resistance determinant detected among the selected isolates was qacEΔ1 (10.00%. A 23.30 of the selected isolates were able to grow on media containing 32 mM copper sulfate, and 46.60% on 8 mM zinc chloride. The metal resistance genes pcoA/copA, pcoR, and chrB were detected in 36.66, 6.66, and 13.33% of selected isolates, respectively. Twelve isolates tested positive for

  18. Influence of Changing Rainfall Patterns on the Yield of Rambutan (Nephelium lappaceum L. and Selection of Genotypes in Known Drought-tolerant Fruit Species for Climate Change Adaptation

    Directory of Open Access Journals (Sweden)

    Pablito M. Magdalita

    2015-06-01

    Full Text Available In fruit crop production, rainfall, water stress, temperature, and wind are key variables for success, and the present changes in rainfall patterns could affect the flowering and yield of the rambutan (Nephelium lappaceum L. Other fruit species like macopa (Syzygium samarangense, siniguelas (Spondias purpurea, and native santol or cotton fruit (Sandoricum koetjape remain productive despite extreme climatic changes. This study assessed the influence of rainfall on rambutan yield and evaluated and selected tree genotypes of known drought-tolerant fruit species. Rambutan yield in a selected farm in Calauan, Laguna, Philippines, dropped remarkably from 152.2 kg/tree in 2008 to 8.6 kg/tree in 2009. This reduction could be attributed to the high rainfall in April 2009 at 334.4 mm, and possibly other environmental factors like temperature, relative humidity, solar radiation, and strong wind. Furthermore, wet months in 2009 also inhibited the flowering of rambutan. However, a low yield obtained in 2010 at 45.5 kg/tree could be partly attributed to the very low rainfall in May 2010 at only 9.1 mm. On the other hand, in relation to changing climate, selection of tree genotypes for use as varieties in known drought- and flood-tolerant fruit species based on important fruit qualities like sweetness, juiciness, and high edible portion was done. Among 103 macopa genotypes, Mc-13, 43, and 91 were selected and the best (i.e. , Mc-13 had sweet (7.15 °Brix and crispy fruits weighing 49.44 g, creamy white (RHCC 155 A, and had high edible portion (EP, 93.22%. Among 114 siniguelas genotypes, Sg-41, 42 and 105 were selected and the best selection (i.e., Sg-41, had sweet (12.50 °Brix and juicy fruit weighing 20.42 g, ruby red (RHCC 59 A, and had high EP (83.27%. Among 101 native santol genotypes, Sn-47, 59, and 74 were selected and the best selection (i.e. , Sn-59 had relatively sweet (5.56 °Brix and juicy fruits weighing 51.96 g, maize yellow (RHCC 21 B, and had

  19. Pharmacokinetics, Pharmacodynamics, Tolerability, and Food Effect of Cenerimod, a Selective S1P₁ Receptor Modulator in Healthy Subjects.

    Science.gov (United States)

    Juif, Pierre-Eric; Baldoni, Daniela; Reyes, Maribel; Wilbraham, Darren; Febbraro, Salvatore; Vaclavkova, Andrea; Hoch, Matthias; Dingemanse, Jasper

    2017-12-06

    The pharmacokinetics, pharmacodynamics, tolerability, and food effect of cenerimod, a potent sphingosine-1-phosphate subtype 1 receptor modulator, were investigated in three sub-studies. Two double-blind, placebo-controlled, randomised studies in healthy male subjects were performed. Cenerimod was administered either as single dose (1, 3, 10 or 25 mg; Study 1) or once daily for 35 days (0.5, 1, 2 or 4 mg; Study 2). A two-period cross-over, open-label study was performed to assess the food effect (1 mg, Study 3). The pharmacokinetic profile of cenerimod was characterised by a t max of 5.0-6.2 h. Terminal half-life after single and multiple doses ranged from 170 to 199 h and 283 to 539 h, respectively. Food had no relevant effect on the pharmacokinetics of cenerimod. A dose-dependent decrease in lymphocyte count was observed after initiation of cenerimod and reached a plateau (maximum change from baseline: -64%) after 20-23 days of treatment. Lymphocyte counts returned to baseline values at end-of-study examination. One serious adverse event of circulatory collapse (25 mg dose group, maximum tolerated dose: 10 mg) and adverse events of mild-to-moderate intensity were reported. Treatment initiation was associated with transient decreases in heart rate and blood pressure at doses >1 and ≥10 mg, respectively.

  20. In vitro and in vivo pharmacological profile of UFP-512, a novel selective delta-opioid receptor agonist; correlations between desensitization and tolerance.

    Science.gov (United States)

    Aguila, B; Coulbault, L; Boulouard, M; Léveillé, F; Davis, A; Tóth, G; Borsodi, A; Balboni, G; Salvadori, S; Jauzac, P; Allouche, S

    2007-12-01

    Delta-opioid receptors (DOP receptors) could represent a novel target in the treatment of depressive disorders. To explore this new field of interest, the development of highly selective DOP receptor agonists is essential. UFP-512 [H-Dmt-Tic-NH-CH(CH2-COOH)-Bid], was recently shown to behave in vitro as a selective and potent DOP receptor agonist and to promote antidepressant- and anxiolytic-like effects in vivo (Vergura et al., 2007). Here, we have characterized the pharmacological properties of UFP-512 and established a link between desensitization and tolerance. Studies were performed in the human neuroblastoma SK-N-BE cells to establish i) binding parameters for UFP-512 ii) signalling pathways activated after acute and chronic treatment iii) regulation (phosphorylation and trafficking) of human DOP (hDOP) receptors after sustained activation by UFP-512. In vivo, we studied UFP-512-induced antidepressant-like effects after acute or chronic treatment in the mouse forced swimming test. In vitro, UFP-512 was a high affinity agonist for DOP receptors. While UFP-512 induced marked phosphorylation of DOP receptors on Ser363, we observed a low desensitization of the cAMP pathway, associated with receptor endocytosis and recycling without any reduction on extracellular signal-regulated protein kinase 1/2 activation. In vivo, acute administration of UFP-512 produced an antidepressant-like effect, without any sign of tolerance after chronic administration. There was a correlation between weak desensitization, significant internalization and recycling of the human DOP receptors and lack of tolerance to UFP-512. This suggests that this compound would be a promising drug prototype for exploring innovative treatments for mood disorders.

  1. The Heterogeneity of Mutational Tolerance in a Protein is Dependent on the Strength of Selective Pressure Correlating with Sectors of Co-evolving Residues

    Science.gov (United States)

    Stiffler, Michael; Ranganathan, Rama

    2011-03-01

    Proteins are capable of tolerating mutations at many positions while still maintaining fold and function. Previous studies have failed to consider how tolerance to random mutagenesis might depend on the strength of selective pressure. To examine this, we measured the fitness of every single point mutation of TEM-1 beta-lactamase across a range of ampicillin concentrations utilizing a novel application of deep-sequencing. We found that the relative mutational robustness between positions varied considerably with respect to ampicillin concentration: at a low ampicillin concentration only a few positions are intolerant of mutations, while at a higher ampicillin concentration many additional positions are as equally intolerant of mutations. Using an analytic method termed statistical coupling analysis (SCA) to measure the co-variation between all positions in a sequence alignment of beta-lactamases revealed sectors of co-evolving positions associated with groups of residues having increased sensitivity to mutagenesis at either low or high ampicillin concentrations. Our findings suggest that nature has ``designed'' proteins to be robust to random mutagenesis by loading the constraints for fitness on discrete networks of co-evolving positions depending on the strength of selective pressure.

  2. A 15-week experimental exposure of pigs to airborne dust with added endotoxin in a continuous flow exposure chamber.

    OpenAIRE

    Jolie, R; Bäckström, L; Olson, L; Chase, C

    1999-01-01

    The purpose of this study was to evaluate the effect of longterm exposure to airborne dust and endotoxin on the respiratory system of pigs. A continuous flow exposure chamber was built for the purpose of exposing pigs to selected airborne contaminants. Pigs (n = 6) were exposed to a combination of a very fine corn/soybean meal (40.6 mg/m3) with added lipopolysaccharide (LPS; 12.4 microg/m3) for 8 h/d over 5 d for 15 wk (75 d of exposure). Control pigs (n = 6) were housed in a room with minima...

  3. Evaluation of Diversity Based on Morphological Variabilities and ISSR Molecular Markers in Iranian Cynodon dactylon (L.) Pers. Accessions to Select and Introduce Cold-Tolerant Genotypes.

    Science.gov (United States)

    Akbari, M; Salehi, H; Niazi, A

    2018-04-01

    The main goals of the present study were to screen Iranian common bermudagrasses to find cold-tolerant accessions and evaluate their genetic and morphological variabilities. In this study, 49 accessions were collected from 18 provinces of Iran. One foreign cultivar of common bermudagrass was used as control. Morphological variation was evaluated based on 14 morphological traits to give information about taxonomic position of Iranian common bermudagrass. Data from morphological traits were evaluated to categorize all accessions as either cold sensitive or tolerant using hierarchical clustering with Ward's method in SPSS software. Inter-Simple Sequence Repeat (ISSR) primers were employed to evaluate genetic variability of accessions. The results of our taxonomic investigation support the existence of two varieties of Cynodon dactylon in Iran: var. dactylon (hairless plant) and var. villosous (plant with hairs at leaf underside and/or upper side surfaces or exterior surfaces of sheath). All 15 primers amplified and gave clear and highly reproducible DNA fragments. In total, 152 fragments were produced, of which 144 (94.73%) being polymorphic. The polymorphic information content (PIC) values ranged from 0.700 to 0.928. The average PIC value obtained with 15 ISSR primers was 0.800, which shows that all primers were informative. Probability identity (PI) and discriminating power between all primers ranged from 0.029 to 0.185 and 0.815 to 0.971, respectively. Genetic data were converted into a binary data matrix. NTSYS software was used for data analysis. Clustering was done by the unweighted pair-group method with arithmetic averages and principle coordinate analysis, separated the accessions into six main clusters. According to both morphological and genetic diversity investigations of accessions, they can be clustered into three groups: cold sensitive, cold semi-tolerant, and cold tolerant. The most cold-tolerant accessions were: Taft, Malayear, Gorgan, Safashahr

  4. Optimal body balance disturbance tolerance skills as a methodological basis for selection of firefighters to solve difficult rescue tasks

    Directory of Open Access Journals (Sweden)

    Władysław Jagiełło

    2014-03-01

    Full Text Available The aim of this study is the methodology of optimal choice of firefighters to solve difficult rescue tasks. 27 firefighters were analyzed: aged from 22–50 years of age, and with 2–27 years of work experience. Body balance disturbance tolerance skills (BBDTS measured by the ‘Rotational Test’ (RT and time of transition (back and forth on a 4 meter beam located 3 meters above the ground, was the criterion for simulation of a rescue task (SRT. RT and SRT were carried out first in a sports tracksuit and then in protective clothing. A total of 4 results of the RT and SRT is the substantive base of the 4 rankings. The correlation of the RT and SRT results with 3 criteria for estimating BBDTS and 2 categories ranged from 0.478 (p<0.01 – 0.884 (p<0.01 and the results of SRT 0.911 (p<0.01. The basic ranking very highly correlated indicators of SRT (0.860 and 0.844, while the 6 indicators of RT only 2 (0.396 and 0.381; p<0.05. There was no correlation between the results of the RT and SRT, but there was an important partial correlation of these variables, but only then was the effect stabilized. The Rotational Test is a simple and easy to use tool for measuring body balance disturbance tolerance skills. However, the BBDTS typology is an accurate criteria for forecasting on this basis, including the results of accurate motor simulations, and the periodic ability of firefighters to solve the most difficult rescue tasks.

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

  7. Origin of endotoxemia influences the metabolic response to endotoxin in dogs

    NARCIS (Netherlands)

    Moeniralam, H. S.; Bemelman, W. A.; Romijn, J. A.; Endert, E.; Ackermans, M. T.; van Lanschot, J. J.; Hermsen, R. C.; Sauerwein, H. P.

    1997-01-01

    Different routes of endotoxin administration have been used to mimic inflammatory and metabolic responses observed during sepsis. Because the origin of endotoxemia may affect the reactions to endotoxin, we compared the induction of tumor necrosis factor (TNF), interleukin-6 (IL-6), hormones, and

  8. Detoxifying Escherichia coli for endotoxin-free production of recombinant proteins.

    Science.gov (United States)

    Mamat, Uwe; Wilke, Kathleen; Bramhill, David; Schromm, Andra Beate; Lindner, Buko; Kohl, Thomas Andreas; Corchero, José Luis; Villaverde, Antonio; Schaffer, Lana; Head, Steven Robert; Souvignier, Chad; Meredith, Timothy Charles; Woodard, Ronald Wesley

    2015-04-16

    Lipopolysaccharide (LPS), also referred to as endotoxin, is the major constituent of the outer leaflet of the outer membrane of virtually all Gram-negative bacteria. The lipid A moiety, which anchors the LPS molecule to the outer membrane, acts as a potent agonist for Toll-like receptor 4/myeloid differentiation factor 2-mediated pro-inflammatory activity in mammals and, thus, represents the endotoxic principle of LPS. Recombinant proteins, commonly manufactured in Escherichia coli, are generally contaminated with endotoxin. Removal of bacterial endotoxin from recombinant therapeutic proteins is a challenging and expensive process that has been necessary to ensure the safety of the final product. As an alternative strategy for common endotoxin removal methods, we have developed a series of E. coli strains that are able to grow and express recombinant proteins with the endotoxin precursor lipid IVA as the only LPS-related molecule in their outer membranes. Lipid IVA does not trigger an endotoxic response in humans typical of bacterial LPS chemotypes. Hence the engineered cells themselves, and the purified proteins expressed within these cells display extremely low endotoxin levels. This paper describes the preparation and characterization of endotoxin-free E. coli strains, and demonstrates the direct production of recombinant proteins with negligible endotoxin contamination.

  9. A method for in Vivo radiolabeling Bacillus thuringiensis native δ-endotoxin crystals

    Science.gov (United States)

    C. Noah Koller; Leah S. Bauer; Robert M. Hollingworth

    1995-01-01

    The entomocidal CryIIIA δ-endotoxin protein of Bacillus thuringiensis var. tenebrionis is distinctive in chemistry and host range. In contrast to other δ-endotoxins, the CryIIIA parasporal crystals are toxic within the acidic midgut environment of several coleopteran species, particularly those in the family...

  10. 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....

  11. 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.

  12. 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.

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

    Directory of Open Access Journals (Sweden)

    Wilmar Correa

    2017-12-01

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

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

    International Nuclear Information System (INIS)

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

    1987-01-01

    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 3 H-thymidine incorporation and direct cell count. Approximately 200 micrograms/ml endotoxin caused a 50% reduction in 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

  16. Personal exposure to dust and endotoxin in Robusta and Arabica coffee processing factories in Tanzania.

    Science.gov (United States)

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

    2013-03-01

    Endotoxin exposure associated with organic dust exposure has been studied in several industries. Coffee cherries that are dried directly after harvest may differ in dust and endotoxin emissions to those that are peeled and washed before drying. The aim of this study was to measure personal total dust and endotoxin levels and to evaluate their determinants of exposure in coffee processing factories. Using Sidekick Casella pumps at a flow rate of 2l/min, total dust levels were measured in the workers' breathing zone throughout the shift. Endotoxin was analyzed using the kinetic chromogenic Limulus amebocyte lysate assay. Separate linear mixed-effects models were used to evaluate exposure determinants for dust and endotoxin. Total dust and endotoxin exposure were significantly higher in Robusta than in Arabica coffee factories (geometric mean 3.41 mg/m(3) and 10 800 EU/m(3) versus 2.10 mg/m(3) and 1400 EU/m(3), respectively). Dry pre-processed coffee and differences in work tasks explained 30% of the total variance for total dust and 71% of the variance for endotoxin exposure. High exposure in Robusta processing is associated with the dry pre-processing method used after harvest. Dust and endotoxin exposure is high, in particular when processing dry pre-processed coffee. Minimization of dust emissions and use of efficient dust exhaust systems are important to prevent the development of respiratory system impairment in workers.

  17. Exhaled nitric oxide in endotoxin-exposed adults: effect modification by smoking and atopy.

    NARCIS (Netherlands)

    Smit, L.A.; Heederik, D.J.J.; Doekes, G.; Wouters, I.M.

    2009-01-01

    OBJECTIVES: Occupational exposure to endotoxin is associated with non-allergic asthma and other airway inflammatory reactions. Little is known about the role of mucosal nitric oxide (NO) production in endotoxin-induced airway inflammation. The objective was to explore exposure-response relationships

  18. 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

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

    Science.gov (United States)

    Ding, Jeak Ling; Ho, Bow

    2010-01-01

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

  20. Efficacy and tolerability of tropisetron in primary fibromyalgia--a highly selective and competitive 5-HT3 receptor antagonist. German Fibromyalgia Study Group.

    Science.gov (United States)

    Färber, L; Stratz, T; Brückle, W; Späth, M; Pongratz, D; Lautenschläger, J; Kötter, I; Zöller, B; Peter, H H; Neeck, G; Alten, R; Müller, W

    2000-01-01

    Based on a potential role for serotonin receptors in fibromyalgia, we investigated the efficacy and tolerability of treatment with tropisetron, a highly selective, competitive inhibitor of the 5-HT3 receptor. In this prospective, multicenter, double-blind, parallel-group, dose-finding study, 418 patients suffering from primary fibromyalgia (ACR criteria) were randomly assigned to receive either placebo, 5 mg, 10 mg or 15 mg tropisetron once daily, respectively. The duration of treatment was 10 days. The clinical response was measured by changes in pain-score, visual analog scale (VAS), and the number of painful tender-points. Treatment with 5 mg tropisetron resulted in a significantly higher response rate (39.2%) when compared with placebo (26.2%) (p=0.033). The absolute reduction in pain-score was -13.5% for 5 mg tropisetron, -13.0% for 10 mg tropisetron, and -6.3% for placebo (pVAS, while a clear trend in terms of clinical benefit was seen with 10 mg tropisetron. The number of painful tender-points was also reduced significantly (p=0.002) in the 5 mg tropisetron group. Of interest, during the 12-month follow-up period, pain intensity of responders on 5 mg and 10 mg tropisetron was still markedly below baseline. The treatment was well tolerated, with gastro-intestinal complaints being the most frequently reported side effects, in keeping with the known safety profile for 5-HT3 receptor antagonists. This study demonstrates the efficacy of short-term treatment with 5 mg tropisetron once daily in primary fibromyalgia. Treatment was well tolerated and prolonged clinical benefits were seen.

  1. 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)

  2. 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.

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

    Directory of Open Access Journals (Sweden)

    Seng Koon Lim

    2014-06-01

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

  4. Clinical safety, tolerability, pharmacokinetics and effects on urinary electrolyte excretion of AZD997, a novel, selective mineralocorticoid receptor modulator.

    Science.gov (United States)

    Erlandsson, Fredrik; Albayaty, Muna; Chialda, Ligia; Ericsson, Hans; Amilon, Carl; Nelander, Karin; Jansson-Löfmark, Rasmus; Wernevik, Linda; Kjaer, Magnus; Bamberg, Krister; Hartleib-Geschwindner, Judith

    2018-02-21

    AZD9977 is the first MR modulator in clinical development exerting similar organ protection as eplerenone with minimal urinary electrolyte effects in pre-clinical studies. The aim was to perform the initial clinical assessment of AZD9977. A first in man trial explored doses from 5-1200 mg. To study effects on urinary electrolyte excretion an additional randomized placebo controlled cross-over 4 period clinical trial was performed. 23 healthy volunteers were administered fludrocortisone alone or in combination with AZD9977, eplerenone or both. AZD9977/eplerenone combination was given to assess if AZD9977 can attenuate eplerenone induced natriuresis. AZD9977 at doses from 5-1200 mg were safe and well tolerated and pharmacokinetics were compatible with further development. AZD9977 exhibited similar effects on urinary ln [Na + ]/ [K + ] as eplerenone when using fludrocortisone as MR agonist, and the combination had an additive effect on ln [Na + K + ]. The results in man contradict the results in rodent models driven by aldosterone, in which AZD9977 has minimal electrolyte effects. Future clinical studies with AZD9977 should be performed in presence of endogenous or exogenous aldosterone to assess potential benefit of AZD9977 in patients. This article is protected by copyright. All rights reserved.

  5. 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.

  6. Toxoplasma gondii Oral Infection Induces Intestinal Inflammation and Retinochoroiditis in Mice Genetically Selected for Immune Oral Tolerance Resistance

    Science.gov (United States)

    Dias, Raul Ramos Furtado; de Carvalho, Eulógio Carlos Queiroz; Leite, Carla Cristina da Silva; Tedesco, Roberto Carlos; Calabrese, Katia da Silva; Silva, Antonio Carlos; DaMatta, Renato Augusto; de Fatima Sarro-Silva, Maria

    2014-01-01

    Toxoplasmosis is a worldwide disease with most of the infections originating through the oral route and generates various pathological manifestations, ranging from meningoencephalitis to retinochoroiditis and inflammatory bowel disease. Animal models for these pathologies are scarce and have limitations. We evaluated the outcome of Toxoplasma gondii oral infection with 50 or 100 cysts of the ME-49 strain in two lines of mice with extreme phenotypes of susceptibility (TS) or resistance (TR) to immune oral tolerance. Therefore, the aim of this study was to evaluate the behaviour of TS and TR mice, orally infected by T. gondii, and determine its value as a model for inflammatory diseases study. Mortality during the acute stage of the infection for TR was 50% for both dosages, while 10 and 40% of the TS died after infection with these respective dosages. In the chronic stage, the remaining TS succumbed while TR survived for 90 days. The TS displayed higher parasite load with lower intestinal inflammation and cellular proliferation, notwithstanding myocarditis, pneumonitis and meningoencephalitis. TR presented massive necrosis of villi and crypt, comparable to inflammatory bowel disease, with infiltration of lymphoid cells in the lamina propria of the intestines. Also, TR mice infected with 100 cysts presented intense cellular infiltrate within the photoreceptor layer of the eyes, changes in disposition and morphology of the retina cell layers and retinochoroiditis. During the infection, high levels of IL-6 were detected in the serum of TS mice and TR mice presented high amounts of IFN-γ and TNF-α. Both mice lineages developed different disease outcomes, but it is emphasized that TR and TS mice presented acute and chronic stages of the infection, demonstrating that the two lineages offer an attractive model for studying toxoplasmosis. PMID:25437299

  7. Toxoplasma gondii oral infection induces intestinal inflammation and retinochoroiditis in mice genetically selected for immune oral tolerance resistance.

    Directory of Open Access Journals (Sweden)

    Raul Ramos Furtado Dias

    Full Text Available Toxoplasmosis is a worldwide disease with most of the infections originating through the oral route and generates various pathological manifestations, ranging from meningoencephalitis to retinochoroiditis and inflammatory bowel disease. Animal models for these pathologies are scarce and have limitations. We evaluated the outcome of Toxoplasma gondii oral infection with 50 or 100 cysts of the ME-49 strain in two lines of mice with extreme phenotypes of susceptibility (TS or resistance (TR to immune oral tolerance. Therefore, the aim of this study was to evaluate the behaviour of TS and TR mice, orally infected by T. gondii, and determine its value as a model for inflammatory diseases study. Mortality during the acute stage of the infection for TR was 50% for both dosages, while 10 and 40% of the TS died after infection with these respective dosages. In the chronic stage, the remaining TS succumbed while TR survived for 90 days. The TS displayed higher parasite load with lower intestinal inflammation and cellular proliferation, notwithstanding myocarditis, pneumonitis and meningoencephalitis. TR presented massive necrosis of villi and crypt, comparable to inflammatory bowel disease, with infiltration of lymphoid cells in the lamina propria of the intestines. Also, TR mice infected with 100 cysts presented intense cellular infiltrate within the photoreceptor layer of the eyes, changes in disposition and morphology of the retina cell layers and retinochoroiditis. During the infection, high levels of IL-6 were detected in the serum of TS mice and TR mice presented high amounts of IFN-γ and TNF-α. Both mice lineages developed different disease outcomes, but it is emphasized that TR and TS mice presented acute and chronic stages of the infection, demonstrating that the two lineages offer an attractive model for studying toxoplasmosis.

  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. Escin attenuates acute lung injury induced by endotoxin in mice.

    Science.gov (United States)

    Xin, Wenyu; Zhang, Leiming; Fan, Huaying; Jiang, Na; Wang, Tian; Fu, Fenghua

    2011-01-18

    Endotoxin causes multiple organ dysfunctions, including acute lung injury (ALI). The current therapeutic strategies for endotoxemia are designed to neutralize one or more of the inflammatory mediators. Accumulating experimental evidence suggests that escin exerts anti-inflammatory and anti-edematous effects. The aim of this study was to evaluate the effect of escin on ALI induced by endotoxin in mice. ALI was induced by injection of lipopolysaccharide (LPS) intravenously. The mice were given dexamethasone or escin before injection of LPS. The mortality rate was recorded. Tumor necrosis factor-α (TNF-α), interleukin 1β (IL-1β) and nitric oxide (NO) were measured. Pulmonary superoxide dismutase (SOD), glutathione peroxidase (GPx) activity, glutathione (GSH), malondialdehyde (MDA) contents, and myeloperoxidase (MPO) activity were also determined. The expression of glucocorticoid receptor (GR) level was detected by Western blotting. Pretreatment with escin could decrease the mortality rate, attenuate lung injury resulted from LPS, down-regulate the level of the inflammation mediators, including NO, TNF-α, and IL-1β, enhance the endogenous antioxidant capacity, and up-regulating the GR expression in lung. The results suggest that escin may have potent protective effect on the LPS-induced ALI by inhibiting of the inflammatory response, and its mechanism involves in up-regulating the GR and enhancing the endogenous antioxidant capacity. Copyright © 2010 Elsevier B.V. All rights reserved.

  10. Quantifying Dustiness, Specific Allergens, and Endotoxin in Bulk Soya Imports

    Directory of Open Access Journals (Sweden)

    Howard J. Mason

    2017-11-01

    Full Text Available Soya is an important bulk agricultural product often transported by sea as chipped beans and/or the bean husks after pelletisation. There are proven allergens in both forms. Bulk handling of soya imports can generate air pollution containing dust, allergens, and pyrogens, posing health risks to dockside workers and surrounding populations. Using an International Organization for Standardization (ISO standardised rotating drum dustiness test in seven imported soya bulks, we compared the generated levels of dust and two major soya allergens in three particle sizes related to respiratory health. Extractable levels of allergen and endotoxin from the bulks showed 30–60 fold differences, with levels of one allergen (hydrophobic seed protein and endotoxin higher in husk. The generated levels of dust and allergens in the three particle sizes also showed very wide variations between bulks, with aerolysed levels of allergen influenced by both the inherent dustiness and the extractable allergen in each bulk. Percentage allergen aerolysed from pelletized husk—often assumed to be of low dustiness—after transportation was not lower than that from chipped beans. Thus, not all soya bulks pose the same inhalation health risk and reinforces the importance of controlling dust generation from handling all soya bulk to as low as reasonably practicable.

  11. Detection of microorganisms, endotoxins and aluminum in mobile dialysis services

    Directory of Open Access Journals (Sweden)

    Priscila Rodrigues de Jesus

    2017-11-01

    Full Text Available Hemodialysis water and dialysates are fundamental in the treatment of kidney disease. During just one hemodialysis session, 120 liters of dialysate are consumed. Thus, it is essential that the parameters of chemical and microbiological quality of the fluids used in dialysis systems are carefully observed. In this study, water samples were collected at 12 hospitals in the state of Rio de Janeiro. The samples were obtained at three points of fluid reservoirs: pre-, post-osmosis and dialysis solution. After collection, colony forming units (CFU, total coliforms and Escherichia coli 100 mL-1 were quantified. Later, isolated colonies and endotoxin content were identified by biochemical assays. Data about total aluminum levels per sample (mg L-1 were also obtained. Samples of all mobile dialysis services and points of collection were contaminated above the levels set out by national laws, in particular by Pseudomonas aeruginosa. Endotoxin levels were also above the recommended by current legislation (> 0.25 EU mL-1. Only three samples contained detectable levels of aluminum, which were found to be above the recommended values for the corresponding resolution (0.01 mg L-1. Finally, there were no observable amounts of total coliforms and E. coli 100 mL-1 sample. The data from this study are an important step forward in the standardization and control of chemical/microbiological quality of mobile dialysis services.

  12. Measurement of operative plasma endotoxin levels in jaundiced and non-jaundiced patients.

    Science.gov (United States)

    Pain, J A; Bailey, M E

    1987-01-01

    A study of portal plasma endotoxin levels was performed using a chromogenic limulus amoebocyte lysate (LAL) assay. The assay proved sensitive and reproducible. In only 1 of 25 healthy subjects was the systemic plasma endotoxin level above 100 pg/ml (equivalent Escherichia coli 0111B4). In 30 non-jaundiced patients undergoing surgery the mean (+SEM) portal plasma endotoxin level (60 + 9 pg/ml) was significantly higher (p less than 0.05) than the mean level in the systemic blood (46 + 6 pg/ml), supporting the concept of endotoxin absorption from the intestine into the portal blood. In 20 patients with obstructive jaundice undergoing surgery 42% of portal, 45% of inferior mesenteric and 35% of systemic venous plasma endotoxin levels were above 100 pg/ml. There were significantly higher levels in the portal (p less than 0.05) and inferior mesenteric (p less than 0.05) compared with the systemic blood. Neither the presence of malignancy nor the duration of surgery appeared to influence endotoxin absorption. The significance of raised plasma endotoxin levels in obstructive jaundice is discussed.

  13. Importance of the personal endotoxin cloud in school-age children with asthma.

    Science.gov (United States)

    Rabinovitch, Nathan; Liu, Andrew H; Zhang, Lening; Rodes, Charles E; Foarde, Karin; Dutton, Steven J; Murphy, James R; Gelfand, Erwin W

    2005-11-01

    A number of studies have observed associations between the amount of endotoxin in urban dust and chronic asthma severity, but a direct relationship between personal exposure to household endotoxin and acute asthma worsening has not yet been defined. We sought to investigate the relationship between day-to-day changes in personal endotoxin exposure and asthma severity. In the winter and spring of 1999 through 2000, endotoxin exposures were monitored in asthmatic schoolchildren by using portable, as opposed to stationary, monitors designed to measure inhalable and respirable particulate matter less than or equal to 2.5 and 10 microm in diameter. Children were followed with daily measurements of FEV(1) and asthma symptoms. Over a 24-hour period, median daily personal endotoxin exposures ranged from 0.08 EU/m(3) (measured at a particulate matter size range Personal exposures were significantly (P personal activities might be better correlated with disease severity. Increases in personal endotoxin exposures were associated with decreased FEV(1) values and increased symptoms. These findings demonstrate the importance of using personal monitoring to both measure and correlate endotoxin exposure with asthma severity.

  14. Endothelin B receptors preserve renal blood flow in a normotensive model of endotoxin-induced acute kidney dysfunction.

    Science.gov (United States)

    Nitescu, Nicoletta; Grimberg, Elisabeth; Ricksten, Sven-Erik; Herlitz, Hans; Guron, Gregor

    2008-03-01

    The aim was to investigate the role of endothelin 1 receptor subtypes in the early renal response to lipopolysaccharide (LPS) during normotensive endotoxemia with acute kidney dysfunction. Endotoxemia was induced in thiobutabarbital-anesthetized rats (n = 9 per group) by infusion of LPS (dosage, 1 mg/kg per hour i.v.). The study groups (1) sham-saline, (2) LPS-saline, (3) LPS-BQ123, (4) LPS-BQ788 and (5) LPS-BQ123 + BQ788 received isotonic saline, the ETA receptor antagonist BQ-123 (dosage, 30 nmol/kg per minute i.v.), and/or the ETB receptor antagonist BQ-788 (dosage, 30 nmol/kg per minute i.v.) before and during 2 h of LPS infusion. Renal clearance measurements, renal blood flow (RBF), and cortical and outer medullary perfusion (laser-Doppler flowmetry) and oxygen tension (Clark-type microelectrodes) were analyzed throughout. Before LPS administration, there were no significant differences between groups in glomerular filtration rate (GFR), RBF, or in cortical (CLDF) and outer medullary perfusion. However, mean arterial pressure (MAP) was elevated in LPS-BQ788 group compared with LPS-BQ123 + BQ788 group (P < 0.05). In saline-treated rats, endotoxin induced an approximate 35% reduction in GFR (P < 0.05), without significant effects on MAP, RBF, or on CLDF and cortical PO2. In addition, LPS increased outer medullary perfusion and PO2 (P < 0.05). The fractional urinary excretion rates of sodium, potassium, and water were not significantly different in LPS-saline group compared with sham-saline group. Neither selective nor combined ETA and ETB receptor blockade improved GFR. In BQ-788-infused rats, endotoxin produced marked reductions in RBF (-18% +/- 4% [P < 0.05]) and CLDF (-18% +/- 2% [P < 0.05]). Similarly, endotoxin decreased RBF (-14% +/- 3% [P < 0.05]) and CLDF (-10% +/- 2% [P < 0.05]) in LPS-BQ123 + BQ788 group. Endotoxin reduced MAP (-22% +/- 4% [P < 0.05]) in BQ-123-treated rats but did not significantly influence MAP in other groups. We conclude that in

  15. 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.

  16. SELEÇÃO PARA TOLERÂNCIA AO ALUMÍNIO EM SOJA TROPICAL SELECTION FOR ALUMINUM TOLERANCE IN TROPICAL SOYBEANS

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    Carlos Roberto Spehar

    2007-09-01

    Full Text Available

    A acidez do solo é fator limitante para a maioria das plantas cultivadas no Cerrado Brasileiro. A toxidez causada por alumínio (Al é especialmente séria na subsuperfície, que permanece ácida após o uso de corretivos, por impedir o crescimento radicular e causar suscetibilidade à seca e desbalanceamento nutricional. Aqui objetivou-se a seleção de genótipos de soja com maior tolerância ao Al, pela associação de experimentos em hidroponia e no campo. Cruzamentos incluindo genótipos selecionados no Cerrado foram realizados. Sementes de indivíduos contrastantes, selecionados em hidroponia na geração F2 pelo crescimento radicular, foram obtidas para avaliação de progênies em F3, no campo, e em F4, novamente em hidroponia. Rendimento de grãos e de biomassa das progênies selecionadas foram superiores aos genitores, no experimento em solo ácido. Esses resultados foram confirmados pelo desempenho em hidroponia, indicando que o método de seleção pode ser empregado com êxito em programas de melhoramento para adaptação de cultivos a condições de acidez subsuperficial do solo.

    PALAVRAS-CHAVE: Acidez sub-superficial; Glycine max; estresse; genótipo; melhoramento de plantas.

    Soil acidity is a limiting factor for most of the cultivated plants in the Brazilian Savannah. Toxicity caused by aluminum (Al is especially serious in the acid subsurface, which remains acidic after soil has been amended, by hindering root growth and causing drought susceptibility and nutritional unbalance. This research aimed at selecting soybean with increased tolerance to Al through association of hydroponics and field experiments. Crosses including savannah adapted genotypes were obtained. Seeds of contrasting individuals, selected in hydroponics at F2 generation for root

  17. Investigating Drought Tolerance in Chickpea Using Genome-Wide Association Mapping and Genomic Selection Based on Whole-Genome Resequencing Data

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    Yongle Li

    2018-02-01

    Full Text Available Drought tolerance is a complex trait that involves numerous genes. Identifying key causal genes or linked molecular markers can facilitate the fast development of drought tolerant varieties. Using a whole-genome resequencing approach, we sequenced 132 chickpea varieties and advanced breeding lines and found more than 144,000 single nucleotide polymorphisms (SNPs. We measured 13 yield and yield-related traits in three drought-prone environments of Western Australia. The genotypic effects were significant for all traits, and many traits showed highly significant correlations, ranging from 0.83 between grain yield and biomass to -0.67 between seed weight and seed emergence rate. To identify candidate genes, the SNP and trait data were incorporated into the SUPER genome-wide association study (GWAS model, a modified version of the linear mixed model. We found that several SNPs from auxin-related genes, including auxin efflux carrier protein (PIN3, p-glycoprotein, and nodulin MtN21/EamA-like transporter, were significantly associated with yield and yield-related traits under drought-prone environments. We identified four genetic regions containing SNPs significantly associated with several different traits, which was an indication of pleiotropic effects. We also investigated the possibility of incorporating the GWAS results into a genomic selection (GS model, which is another approach to deal with complex traits. Compared to using all SNPs, application of the GS model using subsets of SNPs significantly associated with the traits under investigation increased the prediction accuracies of three yield and yield-related traits by more than twofold. This has important implication for implementing GS in plant breeding programs.

  18. Investigating Drought Tolerance in Chickpea Using Genome-Wide Association Mapping and Genomic Selection Based on Whole-Genome Resequencing Data.

    Science.gov (United States)

    Li, Yongle; Ruperao, Pradeep; Batley, Jacqueline; Edwards, David; Khan, Tanveer; Colmer, Timothy D; Pang, Jiayin; Siddique, Kadambot H M; Sutton, Tim

    2018-01-01

    Drought tolerance is a complex trait that involves numerous genes. Identifying key causal genes or linked molecular markers can facilitate the fast development of drought tolerant varieties. Using a whole-genome resequencing approach, we sequenced 132 chickpea varieties and advanced breeding lines and found more than 144,000 single nucleotide polymorphisms (SNPs). We measured 13 yield and yield-related traits in three drought-prone environments of Western Australia. The genotypic effects were significant for all traits, and many traits showed highly significant correlations, ranging from 0.83 between grain yield and biomass to -0.67 between seed weight and seed emergence rate. To identify candidate genes, the SNP and trait data were incorporated into the SUPER genome-wide association study (GWAS) model, a modified version of the linear mixed model. We found that several SNPs from auxin-related genes, including auxin efflux carrier protein (PIN3), p-glycoprotein, and nodulin MtN21/EamA-like transporter, were significantly associated with yield and yield-related traits under drought-prone environments. We identified four genetic regions containing SNPs significantly associated with several different traits, which was an indication of pleiotropic effects. We also investigated the possibility of incorporating the GWAS results into a genomic selection (GS) model, which is another approach to deal with complex traits. Compared to using all SNPs, application of the GS model using subsets of SNPs significantly associated with the traits under investigation increased the prediction accuracies of three yield and yield-related traits by more than twofold. This has important implication for implementing GS in plant breeding programs.

  19. High-protein diet selectively reduces fat mass and improves glucose tolerance in Western-type diet-induced obese rats

    Science.gov (United States)

    Stengel, Andreas; Goebel-Stengel, Miriam; Wang, Lixin; Hu, Eugenia; Karasawa, Hiroshi; Pisegna, Joseph R.

    2013-01-01

    Obesity is an increasing health problem. Because drug treatments are limited, diets remain popular. High-protein diets (HPD) reduce body weight (BW), although the mechanisms are unclear. We investigated physiological mechanisms altered by switching diet induced obesity (DIO) rats from Western-type diet (WTD) to HPD. Male rats were fed standard (SD) or WTD (45% calories from fat). After developing DIO (50% of rats), they were switched to SD (15% calories from protein) or HPD (52% calories from protein) for up to 4 weeks. Food intake (FI), BW, body composition, glucose tolerance, insulin sensitivity, and intestinal hormone plasma levels were monitored. Rats fed WTD showed an increased FI and had a 25% greater BW gain after 9 wk compared with SD (P Diet-induced obese rats switched from WTD to HPD reduced daily FI by 30% on day 1, which lasted to day 9 (−9%) and decreased BW during the 2-wk period compared with SD/SD (P < 0.05). During these 2 wk, WTD/HPD rats lost 72% more fat mass than WTD/SD (P < 0.05), whereas lean mass was unaltered. WTD/HPD rats had lower blood glucose than WTD/SD at 30 min postglucose gavage (P < 0.05). The increase of pancreatic polypeptide and peptide YY during the 2-h dark-phase feeding was higher in WTD/HPD compared with WTD/SD (P < 0.05). These data indicate that HPD reduces BW in WTD rats, which may be related to decreased FI and the selective reduction of fat mass accompanied by improved glucose tolerance, suggesting relevant benefits of HPD in the treatment of obesity. PMID:23883680

  20. Forest tree responses to extreme drought and some biotic events: Towards a selection according to hazard tolerance?

    Science.gov (United States)

    Bréda, Nathalie; Badeau, Vincent

    2008-09-01

    The aim of this paper is to illustrate how some extreme events could affect forest ecosystems. Forest tree response can be analysed using dendroecological methods, as tree-ring widths are strongly controlled by climatic or biotic events. Years with such events induce similar tree responses and are called pointer years. They can result from extreme climatic events like frost, a heat wave, spring water logging, drought or insect damage… Forest tree species showed contrasting responses to climatic hazards, depending on their sensitivity to water shortage or temperature hardening, as illustrated from our dendrochronological database. For foresters, a drought or a pest disease is an extreme event if visible and durable symptoms are induced (leaf discolouration, leaf loss, perennial organs mortality, tree dieback and mortality). These symptoms here are shown, lagging one or several years behind a climatic or biotic event, from forest decline cases in progress since the 2003 drought or attributed to previous severe droughts or defoliations in France. Tree growth or vitality recovery is illustrated, and the functional interpretation of the long lasting memory of trees is discussed. A coupled approach linking dendrochronology and ecophysiology helps in discussing vulnerability of forest stands, and suggests management advices in order to mitigate extreme drought and cope with selective mortality.

  1. Pregnancy promotes tolerance to future offspring by programming selective dysfunction in long-lived maternal T cells.

    Science.gov (United States)

    Barton, Brendan M; Xu, Rong; Wherry, E John; Porrett, Paige M

    2017-04-01

    Fetal antigen available during pregnancy induces the proliferation of maternal T cells. It is unknown, however, whether these antigen-activated T cells differentiate into long-lived memory T cells that are capable of mediating rapid-recall responses to tissue antigens. To test the hypothesis that pregnancy induces an alternative fate in fetal-specific maternal T cells, we used a murine model to track longitudinally fetal-specific T cells in pregnant and postpartum animals and test the response of these cells when challenged with the same antigen during sequential pregnancy or skin transplantation. Fetal-specific CD8 + T cells were robustly primed during pregnancy but failed to acquire robust effector functions. These primed cells persisted long term in postpartum animals, frequently maintained a programmed death 1 (PD-1) + phenotype, and failed to expand or produce cytokines robustly in response to second pregnancy or skin transplantation. However, whereas there was no impact on second pregnancy as a result of the persistence of fetal-primed memory CD8 + T cells in the mother, skin grafts bearing the same antigen were rejected more rapidly. Altogether, our data suggest that fetal antigen exposure during pregnancy induces the differentiation of long-lived maternal CD8 + T cells with context-dependent, selective effector dysfunction. This programmed effector dysfunction provides temporal and systemic restraint of maternal anti-fetal alloreactivity to promote reproductive fitness efficiently, while preserving potentially protective effector T cell responses. © Society for Leukocyte Biology.

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

    Science.gov (United States)

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

    2017-05-01

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

  3. Diet Matters: Endotoxin in the Diet Impacts the Level of Allergic Sensitization in Germ-Free Mice.

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    Martin Schwarzer

    Full Text Available Germ-free animals have been used to define the vital role of commensal bacteria on the maturation of the host immune system. However, the role of bacterial residues in diet in this setting is poorly understood. Here we investigated the effect of bacterial contamination in sterile diet on the level of allergic sensitization in germ-free mice. Sterile grain-based diets ST1 and R03 were tested for the level of bacterial contamination. ST1 contained higher amount of bacterial DNA, approximately ten times more endotoxin, and induced higher, TLR4-dependent, cytokine production in dendritic cells compared to R03. In a germ-free mouse model of sensitization to the major birch pollen allergen Bet v 1, feeding on ST1 for at least two generations was associated with decreased production of allergen-specific IgE and IgG1 antibodies in sera in comparison to R03. Furthermore, reduced levels of allergen-specific and ConA-induced cytokines IL-4, IL-5 and IL-13 accompanied by increased levels of IFN-γ were detected in splenocytes cultures of these mice. Our results show that contamination of experimental diet with bacterial residues, such as endotoxin, significantly affects the development of allergic sensitization in germ-free mice. Therefore, careful selection of sterile food is critical for the outcomes of germ-free or gnotobiotic experimental models of immune-deviated diseases.

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

    Science.gov (United States)

    1992-06-24

    tachycardia, tachypnea, hypertriglyceridemia, thrombocytopenia, metabolic acidosis, acute renal failure, hepatic failure, acute respiratory distress...The symptomatology associated with endotoxemia was first noted in modem times in 1928 by Schwartzman, who first described the systemic disseminated...bilirubin, lactate (which indicates an ameliorating effect on metabolic acidosis), and to bring about an increase in the mean arterial pressure in balxxms

  5. Endotoxin tolerance does not limit mild ischemia-reperfusion injury in humans in vivo.

    NARCIS (Netherlands)

    Draisma, A.; Goeij, M. de; Wouters, C.W.; Riksen, N.P.; Oyen, W.J.G.; Rongen, G.A.P.J.M.; Boerman, O.C.; Deuren, M. van; Hoeven, J.G. van der; Pickkers, P.

    2009-01-01

    Animal studies have shown that previous exposure to lipopolysaccharide (LPS) can limit ischemia-reperfusion injury. We tested whether pretreatment with LPS also protects against ischemia-reperfusion injury in humans in vivo. Fourteen volunteers received bolus injections of incremental dosages of LPS

  6. Endotoxin, Ergosterol, Fungal DNA and Allergens in Dust from Schools in Johor Bahru, Malaysia- Associations with Asthma and Respiratory Infections in Pupils

    Science.gov (United States)

    Norbäck, Dan; Markowicz, Pawel; Cai, Gui-Hong; Hashim, Zailina; Ali, Faridah; Zheng, Yi-Wu; Lai, Xu-Xin; Spangfort, Michael Dho; Larsson, Lennart; Hashim, Jamal Hisham

    2014-01-01

    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. PMID

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

    Science.gov (United States)

    Norbäck, Dan; Markowicz, Pawel; Cai, Gui-Hong; Hashim, Zailina; Ali, Faridah; Zheng, Yi-Wu; Lai, Xu-Xin; Spangfort, Michael Dho; Larsson, Lennart; Hashim, Jamal Hisham

    2014-01-01

    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.

  8. 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.

  9. Effect of air temperature, relative humidity and growth stage on rimsulfuron tolerance in selected field maize hybrids

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    Fuentes Cilia L.

    2002-12-01

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    The effect of temperature (T, relative humidity (RH, and growth stage (GS of six field maize hybrids on the level of tolerance to rimsulfuron [N-((4,6-dimethoxypyrimidin-2-yl aminocarbonyl-3-(ethylsulfonyl-2-pyridincsulfonamide] applied at 20, 40 and 60 g a.i ha-l were evaluated. The hybríds tested are adapted to three

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

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    Sourav P Mukherjee

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

  11. Selection of greenery plants' group tolerant to mineralized human wastes for their inclusion into intrasystem mass exchange of BTLSS

    Science.gov (United States)

    Tikhomirova, Natalia; Tikhomirov, Alexander A.; Kalacheva, Galina; Ushakova, Sofya; Trifonov, Sergey V.; Pavlova, Anastasiya

    2016-07-01

    A possible way solving the inclusion problem of the human liquid wastes containing sodium chloride into intrasystem mass exchange of bio-technical life support system (BTLSS) is selection of such species of greenery plants that can utilize sodium chloride, be edible for a human and have rather a high productivity. Our previous works showed that salt-accumulating halophyte Salicornia europaea L. was a promising candidate for sodium chloride inclusion into BTLSS mass exchange. However, with the aim of creation of more various human diet possibilities a set of greenery plants cultivated by the water culture method was estimated. Based on preliminary experiments the plants Brassica juncea L., Nasturtium officinale R. Br., Lepidium sativum angustifolia L. and Salicornia europaea L. were chosen as the investigation objects. The nutrient solution for greenery plant's cultivation was obtained after harvesting the wheat plants grown on the solution with mineralized human wastes' addition. The results of the first stage of the investigations carried out showed that plants of Brassica juncea and Lepidium sativum angustifolia are unpromising for their inclusion into BTLSS because of a set of physiological characteristics. On the next stage of investigations, an experimental model of closed ecosystem was created. For that purpose the plants of Salicornia europaea and Nasturtium officinale were introduced in the phototroph unit structure. It was determined that there was deficiency of main macronutrients for Salicornia europaea and Nasturtium officinale including sodium for Salicornia europaea. The deficiency had an effect on plants productivity and on carry-over of sodium from solution by Salicornia europaea. Thus in the future experiment it is necessary to carry out desalinization of solution by Salicornia europaea plants. Nasturtium officinale, that is rich in essential fatty acids and contributes into variety of human vegetable diet is planned to be cultivated by the

  12. Research on securing no bacteria and nonfeverish property for disposable medical appliances. Inactivation of endotoxin by Co-60 γ ray

    International Nuclear Information System (INIS)

    Hosobuchi, Kazunari; Tanamoto, Kenichi; Haijima, Yuji.

    1996-01-01

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

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

    International Nuclear Information System (INIS)

    Min, Kyoung-Bok; Min, Jin-Young

    2015-01-01

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

  14. Macrophage subset sensitivity to endotoxin tolerisation by Porphyromonas gingivalis.

    Directory of Open Access Journals (Sweden)

    Andrew D Foey

    Full Text Available Macrophages (MΦs determine oral mucosal responses; mediating tolerance to commensal microbes and food whilst maintaining the capacity to activate immune defences to pathogens. MΦ responses are determined by both differentiation and activation stimuli, giving rise to two distinct subsets; pro-inflammatory M1- and anti-inflammatory/regulatory M2- MΦs. M2-like subsets predominate tolerance induction whereas M1 MΦs predominate in inflammatory pathologies, mediating destructive inflammatory mechanisms, such as those in chronic P.gingivalis (PG periodontal infection. MΦ responses can be suppressed to benefit either the host or the pathogen. Chronic stimulation by bacterial pathogen associated molecular patterns (PAMPs, such as LPS, is well established to induce tolerance. The aim of this study was to investigate the susceptibility of MΦ subsets to suppression by P. gingivalis. CD14(hi and CD14(lo M1- and M2-like MΦs were generated in vitro from the THP-1 monocyte cell line by differentiation with PMA and vitamin D3, respectively. MΦ subsets were pre-treated with heat-killed PG (HKPG and PG-LPS prior to stimulation by bacterial PAMPs. Modulation of inflammation was measured by TNFα, IL-1β, IL-6, IL-10 ELISA and NFκB activation by reporter gene assay. HKPG and PG-LPS differentially suppress PAMP-induced TNFα, IL-6 and IL-10 but fail to suppress IL-1β expression in M1 and M2 MΦs. In addition, P.gingivalis suppressed NFκB activation in CD14(lo and CD14(hi M2 regulatory MΦs and CD14(lo M1 MΦs whereas CD14(hi M1 pro-inflammatory MΦs were refractory to suppression. In conclusion, P.gingivalis selectively tolerises regulatory M2 MΦs with little effect on pro-inflammatory CD14(hi M1 MΦs; differential suppression facilitating immunopathology at the expense of immunity.

  15. Inhalation toxicology models of endotoxin- and bioaerosol-induced inflammation.

    Science.gov (United States)

    Thorne, P S

    2000-11-02

    Inhalation toxicology studies in rodents have proven their usefulness for furthering our understanding of the causal agents, mechanisms, and pathology associated with exposures to environmental endotoxins and bioaerosols. Inhalation animal models are used to determine which components of a mixture are the most important toxicants for inducing the observed adverse outcome. They are used to obtain exposure-response relationships for allergens and pro-inflammatory agents to help elucidate disease mechanisms and contribute quantitative data to the risk assessment process. Inhalation models serve as important adjuncts to epidemiology studies and human exposure studies. They are also useful for establishing phenotype in studies of genetic polymorphisms and disease susceptibility and are widely applied for evaluation of safety and efficacy for potential therapeutic agents. In order to produce reliable data, rigorous exposure chamber design, aerosol generation systems, exposure quantitation and experimental protocols must be utilized.

  16. 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.

  17. Endotoxin-induced inflammation disturbs melatonin secretion in ewe.

    Science.gov (United States)

    Herman, Andrzej Przemysław; Wojtulewicz, Karolina; Bochenek, Joanna; Krawczyńska, Agata; Antushevich, Hanna; Pawlina, Bartosz; Zielińska-Górska, Marlena; Herman, Anna; Romanowicz, Katarzyna; Tomaszewska-Zaremba, Dorota

    2017-12-01

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

  18. High mortality rates occur in copper deficient rats exposed to a normally nonlethal endotoxin treatment

    Energy Technology Data Exchange (ETDEWEB)

    DiSilvestro, R.; Joseph, E.; Yang, F.L. (Ohio State Univ., Columbus (United States))

    1991-03-15

    Endotoxin hepatotoxicity is proposed to occur by processes which could be retarded by 3 copper enzymes: ceruloplasmin, Cu-Zn superoxide dismutase (SOD), and extracellular (EC) SOD. Weanling rats fed low copper for 40 days showed low activity levels of these enzymes, and a very high mortality rate 20 h after endotoxin injection. No rats fed adequate copper died from this treatment. In addition, serum transaminase activities, indicators of liver damage, were elevated by 3 h to a greater extent in the deficient rats than in the adequates. The high susceptibility to endotoxemia in the deficient rats was not associated with low hepatic glutathione, high liver malondialedhyde, nor restricted metallothionein induction 3 h after endotoxin injection. Endotoxin reduced serum EC SOD activities in adequate and deficient rats, but final values were lower in the latter. Studies on roles of specific copper enzymes in resistance to endotoxemia are currently underway.

  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. The role of endotoxin in the pathogenesis of acute bovine laminitis.

    Science.gov (United States)

    Boosman, R; Mutsaers, C W; Klarenbeek, A

    1991-07-01

    To study the possible role of endotoxin in the pathogenesis of bovine laminitis, local and systemic injections of endotoxin (E. coli 0111 B4) with different doses were given to three groups of four cows each. Clinical and haematologic parameters indicated an acute-phase response, including positive plasma ethanol gelation (soluble fibrin), the occurrence of fibrin degradation products and decreased thrombocyte counts. Local Shwartzman reactions were not evoked. Clinical examination of the claws and the gait of the animals revealed no signs of laminitis. However, on histopathological examination of the claw corium signs of laminitis such as vacuolisation of the Stratum basale, lymphocyte and leucocyte infiltration and thrombosis were found. These results indicate that endotoxin indeed may be involved in the pathogenesis of laminitis. For the development of a clinical acute laminitis model in cattle either another dosage, other toxins or factors in addition to the endotoxin used in this experiment are needed.

  1. Effect of saccharides on growth, sporulation rate and δ-endotoxin ...

    African Journals Online (AJOL)

    sucrose, lactose and maltose) as carbon sources on growth and sporulation rate of Bacillus thuringiensis MPK13 as well as δ-endotoxin production was investigated using 400 ml shake flasks culture. Substantially high growth and sporulation ...

  2. 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.

  3. 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.

  4. [Removal of oral Prevotella intermedia Endotoxin by octyl phenyl polyoxyethylene ether extraction method].

    Science.gov (United States)

    Wang, Ai-wu; Liu, Yan; Hu, Kong-xin; Cheng, Qian

    2012-12-01

    To investigate an effective purification method for removing endotoxin from Prevotella intermedia. The main protein ingredients of bacteria prepared from ammonium sulfate precipitation were further treated with octyl phenyl polyoxyethylene ether (Triton X-114), and then processed at 4°C, 37°C and 25°C. The obtained aqueous phase after at least two more cycle repeated operations was assayed for endotoxin by Western blotting, LAL-clotting method, in vitro cell stimulation and in vivo animal experiments. Western blotting and LAL-clotting method demonstrated that the reduction in endotoxin level was greater than 99.99% and recovery of the proteins after endotoxin removal was greater than 90% with Triton X-114 treatment for 3 cycles. The cytokines expression level was lower in both in vitro cell stimulation and in vivo animal experiments than in untreated group (P Prevotella intermedia.

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

    International Nuclear Information System (INIS)

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

    1987-01-01

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

  6. Investigations on the specificity of the Limulus test for the detection of endotoxin.

    Science.gov (United States)

    Wildfeuer, A; Heymer, B; Schleifer, K H; Haferkamp, O

    1974-11-01

    Lysates obtained from amoebocytes of Limulus polyphemus, the horseshoe crab, showed gel formation after the addition of bacterial endotoxin. In contrast to living gram-negative bacteria, viable gram-positive microorganisms did not cause gelation of lysate. Nevertheless, peptidoglycan isolated from the cell walls of various gram-positive organisms did induce the reaction. However, the activity of peptidoglycan was 1,000 to 400,000 times less than that of Escherichia coli lipopolysaccharide. After exposure to lysozyme, peptidoglycan no longer gelled amoebocyte lysate, therefore apparently excluding endotoxin contamination. Gelation of amoebocyte lysate by endotoxin or peptidoglycan was inhibited by different concentrations of sodium polystyrolsulfonate. Whereas these studies confirm the specificity of the Limulus test for bacterial endotoxins, they also indicate that other substances of bacterial origin should be investigated for their ability to gel amoebocyte lysate.

  7. Chromogenic substrates for horseshoe crab clotting enzyme. Its application for the assay of bacterial endotoxins.

    Science.gov (United States)

    Iwanaga, S; Morita, T; Harada, T; Nakamura, S; Niwa, M; Takada, K; Kimura, T; Sakakibara, S

    1978-01-01

    An endotoxin-activated hemocyte lysate from the horseshoe crab (Tachypleus and Limulus) was found to hydrolyze Bz-Ile-Glu-(gamma-OR)-Gly-Arg-p-nitroanilide (PNA), Bz-Val-Gly-Arg-PNA, Boc-Val-Leu-Gly-Arg-PNA, and Boc-Leu-Gly-Arg-PNA, all of which have the COOH-terminal Gly-Arg sequence. This amidase activity was due to a clottting enzyme contained in the lysate. Furthermore, the amidase activity increased by increasing the concentration of bacterial endotoxin (E. coli, 0111-B4) added to the lysate. Therefore, the measurement of the endotoxin-induced amidase activity made it possible to determine the concentration of the endotoxin.

  8. Isolation and sequencing of the cryIC-like delta endotoxin gene from ...

    African Journals Online (AJOL)

    Isolation and sequencing of the cryIC-like delta endotoxin gene from Egyptian strains of Bacillus thuringiensis toxic against Lepidoptera. ME Elkashef, AM Heikal, MEDM Solliman, MI Kobeasy, HA El-Shemy ...

  9. Identification of Sources of Endotoxin Exposure as Input for Effective Exposure Control Strategies.

    Science.gov (United States)

    van Duuren-Stuurman, Birgit; Gröllers-Mulderij, Mariska; van de Runstraat, Annemieke; Duisterwinkel, Anton; Terwoert, Jeroen; Spaan, Suzanne

    2018-02-13

    Aim of the present study is to investigate the levels of endotoxins on product samples from potatoes, onions, and seeds, representing a relevant part of the agro-food industry in the Netherlands, to gather valuable insights in possibilities for exposure control measures early in the process of industrial processing of these products. Endotoxin levels on 330 products samples from companies representing the potato, onion, and seed (processing) industry (four potato-packaging companies, five potato-processing companies, five onion-packaging companies, and four seed-processing companies) were assessed using the Limulus Amboecyte Lysate (LAL) assay. As variation in growth conditions (type of soil, growth type) and product characteristics (surface roughness, dustiness, size, species) are assumed to influence the level of endotoxin on products, different types, and growth conditions were considered when collecting the samples. Additionally, waste material, rotten products, felt material (used for drying), and process water were collected. A large variation in the endotoxin levels was found on samples of potatoes, onions, and seeds (overall geometric standard deviation 17), in the range between 0.7 EU g-1 to 16400000 EU g-1. The highest geometric mean endotoxin levels were found in plant material (319600 EU g-1), followed by soil material (49100 EU g-1) and the outer side of products (9300 EU g-1), indicating that removal of plant and soil material early in the process would be an effective exposure control strategy. The high levels of endotoxins found in the limited number of samples from rotten onions indicate that these rotten onions should also be removed early in the process. Mean endotoxin levels found in waste material (only available for seed processing) is similar to the level found in soil material, although the range is much larger. On uncleaned seeds, higher endotoxin levels were found than on cleaned seeds, indicating that cleaning processes are important

  10. Intestinal Endotoxins as Co-Factors of Liver Injury in Obstructive Jaundice

    OpenAIRE

    Menteş, B. Bülent; Tatlicioğlu, Ertan; Akyol, Gülen; Uluoğlu, Ömer; Sultan, Nedim; Yilmaz, Erdal; Çelebi, Murat; Taneri, Ferit; Ferahköşe, Zafer

    1996-01-01

    The concept of endotoxin-mediated rather than direct liver injury in biliary obsruction was investigated using the experimental rat model of bile duct ligation (BDL) and small bowel bacterial overgrowth (SBBO). Small identical doses of intravenous endotoxin (bacterial LPS) caused a significantly more severe liver injury in rats with BDL, compared with sham-operated rats, suggesting the possible contribution of LPS in this type of liver damage. BDL was then combined with surgica...

  11. Comparison Between the Polymyxins and Gentamicin in Preventing Endotoxin-Induced Intravascular Coagulation and Leukopenia

    Science.gov (United States)

    Corrigan, James J.; Bell, Bobbie M.

    1971-01-01

    Three antimicrobial agents were evaluated as to their ability to neutralize the toxic effects of endotoxin in rabbits. These consisted of two cyclic polypeptides, polymyxin B sulfate and colymycin M (sodium colistimethate), and an aminoglycoside, gentamicin sulfate. Polymyxin B regularly prevented endotoxin-induced leukopenia, thrombocytopenia, and disseminated intravascular coagulation. Colymycin M had similar activity but was not as effective as polymyxin B. Gentamicin demonstrated no neutralizing ability in this study. PMID:4343409

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

    OpenAIRE

    Mokhtar, Guizani; Naoyuki, Funamizu

    2012-01-01

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

  13. 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...

  14. Branchial challenge of normal subjects with the endotoxin of Enterobacter agglomerans isolated from cotton dust.

    Science.gov (United States)

    Jamison, J P; Lowry, R C

    1986-05-01

    Endotoxin produced by a culture of Enterobacter agglomerans isolated from cotton dust was inhaled by 12 normal subjects. No significant airway constriction was obtained in doses equivalent to those experienced in a workshift in a dusty mill. There was a statistically significant difference between this result and the bronchoconstriction that had occurred after flax dust inhalation in the same subjects. It is suggested that Enterobacter agglomerans endotoxin is not the causative agent of the acute bronchoconstriction that follows inhalation of textile dust.

  15. Branchial challenge of normal subjects with the endotoxin of Enterobacter agglomerans isolated from cotton dust.

    OpenAIRE

    Jamison, J P; Lowry, R C

    1986-01-01

    Endotoxin produced by a culture of Enterobacter agglomerans isolated from cotton dust was inhaled by 12 normal subjects. No significant airway constriction was obtained in doses equivalent to those experienced in a workshift in a dusty mill. There was a statistically significant difference between this result and the bronchoconstriction that had occurred after flax dust inhalation in the same subjects. It is suggested that Enterobacter agglomerans endotoxin is not the causative agent of the a...

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

    Science.gov (United States)

    Kuo, Shiu-Ming

    2016-01-01

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

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

    OpenAIRE

    Mizumura, Hikaru; 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...

  18. A non-chromatographic method for the removal of endotoxins from bacteriophages.

    Science.gov (United States)

    Branston, Steven D; Wright, Jason; Keshavarz-Moore, Eli

    2015-08-01

    The Ff filamentous bacteriophages show potential as a new class of therapeutics, displaying utility in materials science as well as pharmaceutical applications. These phages are produced by the infection of E. coli, a Gram-negative bacterium which unavoidably sheds endotoxins into the extracellular space during growth. Since endotoxin molecules are highly immunoreactive, separation from the phage product is of critical importance, particularly those developed for human therapeutic use. The properties of M13, one of the Ff group, present a purification challenge chiefly because the standard scalable method for endotoxin removal from proteins-anion exchange chromatography-is not applicable due to pI similarity between the particles. This article examines the potential of polyethylene glycol (PEG)-NaCl precipitation as a scalable method for the separation of endotoxins from phage M13. Precipitation of M13 by 2% (w/v) PEG 6 000, 500 mM NaCl reduced endotoxin contamination of the phage product by 88%, but additional precipitation rounds did not maintain this proportional decrease. Dynamic light scattering was subsequently used to determine the effectiveness of a detergent to disassociate endotoxin molecules from M13. As a result, PEG-NaCl precipitation was supplemented with up to 2% (v/v) Triton X-100 to improve separation. A 5.7 log10 reduction in endotoxin concentration was achieved over three rounds of precipitation whilst retaining over 97% of the phage. This method compares favorably with the well-known ATPS (Triton X-114) technique for endotoxin removal from protein solutions. © 2015 Wiley Periodicals, Inc.

  19. Cloricromene, a coumarine derivative, protects against lethal endotoxin shock in rats.

    Science.gov (United States)

    Squadrito, F; Altavilla, D; Campo, G M; Calapai, G; Ioculano, M; Zingarelli, B; Saitta, A; Prosdocimi, M; Caputi, A P

    1992-01-14

    Endotoxin shock was induced in male rats by an intravenous (i.v.) injection of Salmonella enteriditis lipopolysaccharide (LPS; 20 mg/kg i.v.). Survival rate, macrophage and serum tumor necrosis factor (TNF-alpha), mean arterial blood pressure (MAP) and white blood cell count were then evaluated. Furthermore the in vitro effect of cloricromene on peritoneal macrophage phagocytosis and TNF-alpha release by primed peritoneal macrophages was investigated. LPS administration caused animal death (0% survival 24 h after endotoxin challenge), hypotension, marked leukopenia and increased the levels of TNF-alpha in both serum and macrophage supernatants. Cloricromene administration (0.5, 1 and 2 mg/kg i.v. 15 min after endotoxin) protected against LPS-induced lethality (100% survival rate 24 h after endotoxin challenge), reverted LPS-induced hypotension and leukopenia, and decreased TNF-alpha in both serum and macrophage supernatants. Finally, cloricromene, added in vitro to peritoneal macrophages collected from endotoxin-treated rats increased macrophage phagocytosis and reduced TNF-alpha formation by activated mononuclear phagocytes. Our data suggest that cloricromene increases survival rate in endotoxin shock through an inhibition of TNF-alpha production.

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

    Directory of Open Access Journals (Sweden)

    Bożena Szermer-Olearnik

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

  1. Endotoxin concentration in treated water of all hemodialysis units in Greece and inquisition of influencing factors.

    Science.gov (United States)

    Arvanitidou, M; Spaia, S; Askepidis, N; Kanetidis, D; Pazarloglou, M; Katsouyannopoulos, V; Vayonas, G

    1999-01-01

    Mains and purified water samples from all hemodialysis centers in Greece were surveyed to assess hemodialysis water quality and the potential consequences on the dialysis population of any microbial and endotoxin contamination. Total heterotrophic bacteria counts were estimated employing the pour-plate method, the membrane filter technique, for total coliforms, fecal coliforms, enterococci and Pseudomonas spp., and the most probable number method for sulfite-reducing clostridia. Overall 170 paired samples were tested. Endotoxin concentrations were assayed in treated water samples, by the Limulus Amebocyte Lysate method, employing a chromogenic substrate. Endotoxin concentration varied from 0-30 EU/ml and exceeded 5 EU/ml in 30.6% of the centers. Endotoxin contaminated samples showed higher counts of total heterotrophic bacteria, fecal coliforms, enterococci and Pseudomonas spp. than the rest of the samples, though the difference was significant only for total heterotrophic bacteria and enterococci. Additionally, samples with endotoxin contamination derived from the mains water had significantly higher counts of fecal coliforms, enterococci and Pseudomonas spp. Endotoxin concentration was correlated with the presence of a deionization system, counts of fecal coliforms in supply water and counts of total coliforms, fecal coliforms and enterococci in treated water. These results demonstrate that hemodialysis centers in Greece need to monitor and preventively maintain the entire hemodialysis system, in order to ensure renal replacement therapy of good quality.

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

    Science.gov (United States)

    Neun, Barry W; Dobrovolskaia, Marina A

    2011-01-01

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

  3. Removal of endotoxin from deionized water using micromachined silicon nanopore membranes

    International Nuclear Information System (INIS)

    Smith, Ross A; Fissell, William H; Fleischman, Aaron J; Roy, Shuvo; Goldman, Ken; Zorman, Christian A

    2011-01-01

    Endotoxins are lipopolysaccharide components of the cell membrane of Gram-negative bacteria that trigger the body's innate immune system and can cause shock and death. Water for medical therapy, including parenteral and dialysate solutions, must be free of endotoxin. This purity is challenging to achieve as many Gram-negative bacteria are endemic in the environment, and can thrive in harsh, nutrient-poor conditions. Current methods for removing endotoxin include distillation and reverse osmosis, both of which are resource intensive processes. Membranes that present an absolute barrier to macromolecular passage may be capable of delivering pure water for biomedical applications. In this work, endotoxin has been filtered from aqueous solutions using silicon nanopore membranes (SNMs) with monodisperse pore size distributions. SNMs with critical pore sizes between 26 and 49 nm were challenged with solutions of deionized water spiked with endotoxin and with Pseudomonas cepacia. The filtrate produced by the SNM from Pseudomonas-contaminated water had <1.0 endotoxin unit (EU) ml −1 , which meets standards for dialysate purity. This approach suggests a technique for single-step cleanup of heavily contaminated water that may be suitable for field or clinical use

  4. Efficacy and Tolerability of an Inhaled Selective Glucocorticoid Receptor Modulator - AZD5423 - in Chronic Obstructive Pulmonary Disease Patients: Phase II Study Results.

    Science.gov (United States)

    Kuna, Piotr; Aurivillius, Magnus; Jorup, Carin; Prothon, Susanne; Taib, Ziad; Edsbäcker, Staffan

    2017-10-01

    AZD5423 is a novel, inhaled, selective glucocorticoid receptor modulator (SGRM), which in an allergen challenge model in asthma patients improved lung function and airway hyper-reactivity. In the current trial, AZD5423 was for the first time tested in patients with chronic obstructive pulmonary disease (COPD). In this double-blind, randomized and parallel group study, we examined airway and systemic effects of two doses of AZD5423, inhaled via Turbuhaler for 12 weeks, in 353 symptomatic patients with COPD (average pre-bronchodilator forced expiratory volume in one-second (FEV1) at screening was 50-52% of predicted normal). Pre-bronchodilator FEV1 was primary variable, with other lung function parameters plus symptoms and 24-hr plasma cortisol being secondary variables. Plasma concentrations of AZD5423 were also measured. Effects were compared against placebo and a reference glucocorticoid receptor agonist control. Neither AZD5423, at doses which have shown to be efficacious in allergen-induced asthma, nor the reference control, at double the approved dose, had any clinically meaningful effect in the patient population studied in regard to lung function or markers of inflammation. Both GR modulators were well tolerated and did suppress 24-hr cortisol. This study suggests that the selected population of patients with COPD does not respond to treatment with AZD5423 as regards lung function, while showing the expected systemic effects. It cannot be ruled out that a favourable lung function response of AZD5423 can be evoked using another experimental setting and/or within a different population of patients with COPD. © 2017 Nordic Association for the Publication of BCPT (former Nordic Pharmacological Society).

  5. 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.

  6. 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.

  7. Long-term interdisciplinary therapy reduces endotoxin level and insulin resistance in obese adolescents.

    Science.gov (United States)

    Lira, Fábio S; Rosa, Jose C; Pimentel, Gustavo D; Santos, Ronaldo V; Carnier, June; Sanches, Priscila L; de Piano, Aline; de Souza, Claudio T; Tock, Lian; Tufik, Sergio; de Mello, Marco T; Seelaender, Marília; Oller do Nascimento, Claudia M; Oyama, Lila M; Dâmaso, Ana R

    2012-09-18

    The purpose of the present study was to assess the dietary fat intake, glucose, insulin, Homeostasis model assessment for insulin resistance HOMA-IR, and endotoxin levels and correlate them with adipokine serum concentrations in obese adolescents who had been admitted to long-term interdisciplinary weight-loss therapy. The present study was a longitudinal clinical intervention of interdisciplinary therapy. Adolescents (n = 18, aged 15-19 y) with a body mass index > 95th percentile were admitted and evaluated at baseline and again after 1 year of interdisciplinary therapy. We collected blood samples, and IL-6, adiponectin, and endotoxin concentrations were measured by ELISA. Food intake was measured using 3-day diet records. In addition, we assessed glucose and insulin levels as well as the homeostasis model assessment for insulin resistance (HOMA-IR). The most important finding from the present investigation was that the long-term interdisciplinary lifestyle therapy decreased dietary fat intake and endotoxin levels and improved HOMA-IR. We observed positive correlations between dietary fat intake and endotoxin levels, insulin levels, and the HOMA-IR. In addition, endotoxin levels showed positive correlations with IL-6 levels, insulin levels and the HOMA-IR. Interestingly, we observed a negative correlation between serum adiponectin and both dietary fat intake and endotoxin levels. The present results indicate an association between dietary fat intake and endotoxin level, which was highly correlated with a decreased pro-inflammatory state and an improvement in HOMA-IR. In addition, this benefits effect may be associated with an increased adiponectin level, which suggests that the interdisciplinary therapy was effective in improving inflammatory pathways.

  8. Long-term interdisciplinary therapy reduces endotoxin level and insulin resistance in obese adolescents

    Directory of Open Access Journals (Sweden)

    Lira Fábio S

    2012-09-01

    Full Text Available Abstract Aim The purpose of the present study was to assess the dietary fat intake, glucose, insulin, Homeostasis model assessment for insulin resistance HOMA-IR, and endotoxin levels and correlate them with adipokine serum concentrations in obese adolescents who had been admitted to long-term interdisciplinary weight-loss therapy. Design The present study was a longitudinal clinical intervention of interdisciplinary therapy. Adolescents (n = 18, aged 15–19 y with a body mass index > 95th percentile were admitted and evaluated at baseline and again after 1 year of interdisciplinary therapy. We collected blood samples, and IL-6, adiponectin, and endotoxin concentrations were measured by ELISA. Food intake was measured using 3-day diet records. In addition, we assessed glucose and insulin levels as well as the homeostasis model assessment for insulin resistance (HOMA-IR. Results The most important finding from the present investigation was that the long-term interdisciplinary lifestyle therapy decreased dietary fat intake and endotoxin levels and improved HOMA-IR. We observed positive correlations between dietary fat intake and endotoxin levels, insulin levels, and the HOMA-IR. In addition, endotoxin levels showed positive correlations with IL-6 levels, insulin levels and the HOMA-IR. Interestingly, we observed a negative correlation between serum adiponectin and both dietary fat intake and endotoxin levels. Conclusions The present results indicate an association between dietary fat intake and endotoxin level, which was highly correlated with a decreased pro-inflammatory state and an improvement in HOMA-IR. In addition, this benefits effect may be associated with an increased adiponectin level, which suggests that the interdisciplinary therapy was effective in improving inflammatory pathways.

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

    International Nuclear Information System (INIS)

    Fukumori, Neuza Taeko Okasaki

    2008-01-01

    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 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 -1 (Endotoxin Units per milliliter). For validation, a dilution series was performed, a twofold dilution of control standard endotoxin (CSE) from 0.5 to 0.03 EU mL -1 , to confirm the labeled sensitivity of the LAL reagent being tested in sterile and non pyrogenic water, in quadruplicate. The same dilution series was performed with the CSE and the product in the 1:100 dilution factor, in three consecutive batches of each radiopharmaceutical. The products 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 valuable quantitative and

  10. Ultra-low concentrations of naloxone selectively antagonize excitatory effects of morphine on sensory neurons, thereby increasing its antinociceptive potency and attenuating tolerance/dependence during chronic cotreatment.

    Science.gov (United States)

    Crain, S M; Shen, K F

    1995-11-07

    Ultra-low picomolar concentrations of the opioid antagonists naloxone (NLX) and naltrexone (NTX) have remarkably potent antagonist actions on excitatory opioid receptor functions in mouse dorsal root ganglion (DRG) neurons, whereas higher nanomolar concentrations antagonize excitatory and inhibitory opioid functions. Pretreatment of naive nociceptive types of DRG neurons with picomolar concentrations of either antagonist blocks excitatory prolongation of the Ca(2+)-dependent component of the action potential duration (APD) elicited by picomolar-nanomolar morphine and unmasks inhibitory APD shortening. The present study provides a cellular mechanism to account for previous reports that low doses of NLX and NTX paradoxically enhance, instead of attenuate, the analgesic effects of morphine and other opioid agonists. Furthermore, chronic cotreatment of DRG neurons with micromolar morphine plus picomolar NLX or NTX prevents the development of (i) tolerance to the inhibitory APD-shortening effects of high concentrations of morphine and (ii) supersensitivity to the excitatory APD-prolonging effects of nanomolar NLX as well as of ultra-low (femtomolar-picomolar) concentrations of morphine and other opioid agonists. These in vitro studies suggested that ultra-low doses of NLX or NTX that selectively block the excitatory effects of morphine may not only enhance the analgesic potency of morphine and other bimodally acting opioid agonists but also markedly attenuate their dependence liability. Subsequent correlative studies have now demonstrated that cotreatment of mice with morphine plus ultra-low-dose NTX does, in fact, enhance the antinociceptive potency of morphine in tail-flick assays and attenuate development of withdrawal symptoms in chronic, as well as acute, physical dependence assays.

  11. Crafting tolerance

    DEFF Research Database (Denmark)

    Kirchner, Antje; Freitag, Markus; Rapp, Carolin

    2011-01-01

    Ongoing changes in social structures, orientation, and value systems confront us with the growing necessity to address and understand transforming patterns of tolerance as well as specific aspects, such as social tolerance. Based on hierarchical analyses of the latest World Values Survey (2005...... results show that specific institutional qualities, which reduce status anxiety, such as inclusiveness, universality, and fairness, prevail over traditional socio-economic, societal, cultural, and democratic explanations....

  12. Characteristics of Endotoxin-Altering Fractions Derived from Normal Serum III. Isolation and Properties of Horse Serum alpha(2)-Macroglobulin.

    Science.gov (United States)

    Yoshioka, M; Konno, S

    1970-05-01

    The endotoxin-altering activity of fractions isolated from normal horse serum was examined by incubation of Salmonella typhosa strain 0-901 endotoxin (Boivin) in a solution of the fraction, and subsequent quantitation of any diminution in the capacity of endotoxin to be precipitated by specific anti-endotoxin antiserum. The horse serum fraction isolated by precipitation with ammonium sulfate at a concentration between 1.6 and 2.7 m was incubated with Pronase PA and then with trypsin. When this partly digested fraction was passed twice through a Sephadex G-200 column and eluted with 0.2 m tris(hydroxymethyl)aminomethane buffer, most of the endotoxinaltering activity was found in the first protein peak designated F-1a. F-1a was found to be homogeneous and corresponded to an alpha(2)-macroglobulin by the techniques of electrophoresis, immunodiffusion, and ultracentrifugation. Approximately 100-fold more F-1a than endotoxin was needed to reduce the antigenicity of the endotoxin by one-half. Alteration was increased when F-1a was incubated with the endotoxin at acid pH or at 45 C rather than at 37 C and was lost after heating F-1a at 56 C for 30 min. N-ethylmaleimide increased the endotoxin-altering activity of horse serum, F-1a, and human plasma fraction III(0), whereas p-chloromercuribenzoate did not. On the other hand, diazonium-1-H-tetrazole, iodoacetic acid, and benzylchloride suppressed the activity of F-1a. When the interaction of endotoxin and F-1a was examined by immunodiffusion techniques, depolymerization of the endotoxin molecule was indicated. The endotoxin-altering factor of horse serum is discussed in relation to the mechanisms of other known reagents, such as deoxycholate and sodium lauryl sulfate.

  13. Characteristics of Endotoxin-Altering Fractions Derived from Normal Serum III. Isolation and Properties of Horse Serum α2-Macroglobulin

    Science.gov (United States)

    Yoshioka, Morimasa; Konno, Seishi

    1970-01-01

    The endotoxin-altering activity of fractions isolated from normal horse serum was examined by incubation of Salmonella typhosa strain 0-901 endotoxin (Boivin) in a solution of the fraction, and subsequent quantitation of any diminution in the capacity of endotoxin to be precipitated by specific anti-endotoxin antiserum. The horse serum fraction isolated by precipitation with ammonium sulfate at a concentration between 1.6 and 2.7 m was incubated with Pronase PA and then with trypsin. When this partly digested fraction was passed twice through a Sephadex G-200 column and eluted with 0.2 m tris(hydroxymethyl)aminomethane buffer, most of the endotoxinaltering activity was found in the first protein peak designated F-1a. F-1a was found to be homogeneous and corresponded to an α2-macroglobulin by the techniques of electrophoresis, immunodiffusion, and ultracentrifugation. Approximately 100-fold more F-1a than endotoxin was needed to reduce the antigenicity of the endotoxin by one-half. Alteration was increased when F-1a was incubated with the endotoxin at acid pH or at 45 C rather than at 37 C and was lost after heating F-1a at 56 C for 30 min. N-ethylmaleimide increased the endotoxin-altering activity of horse serum, F-1a, and human plasma fraction III0, whereas p-chloromercuribenzoate did not. On the other hand, diazonium-1-H-tetrazole, iodoacetic acid, and benzylchloride suppressed the activity of F-1a. When the interaction of endotoxin and F-1a was examined by immunodiffusion techniques, depolymerization of the endotoxin molecule was indicated. The endotoxin-altering factor of horse serum is discussed in relation to the mechanisms of other known reagents, such as deoxycholate and sodium lauryl sulfate. Images PMID:16557754

  14. Measurement of airborne bacteria and endotoxin generated during dental cleaning.

    Science.gov (United States)

    Dutil, Steve; Meriaux, Anne; de Latremoille, Marie-Chantale; Lazure, Louis; Barbeau, Jean; Duchaine, Caroline

    2009-02-01

    Dynamic dental instruments generate abundant aerosols in the work environment. Dental unit waterlines (DUWL) support a large microbial population and can be a significant source of bioaerosols generated during dental treatments. This study was conducted to characterize bioaerosol generation during dental treatments performed in standardized conditions. Culture-based method (R2A, and blood agar with and without O2) and fluorescence microscopy were used. Dental cleaning procedures were performed in an isolated treatment room with controlled ventilation rate. Andersen microbial samplers were used to collect culturable bioaerosols generated before (baseline), during, and after 2 hr of dental treatments. Inhalable dust samplers were used to measure total bioaerosols content in dental hygienist's and patients' breathing zones. AGI-30 were used for the collection of the endotoxin. The use of fluorescence microscopy in combination with culture demonstrated that dental staff and patients were exposed to up to 1.86 E+05 bacteria/m(3) generated during treatments. Fortunately, bioaerosols returned to baseline within 2 hr after the dental procedures. The small diameter of the aerosols generated (< 1 microm) suggests that the risk of contact between the aerosolized bacteria and the respiratory system of exposed individuals is likely to occur.

  15. 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.

  16. Endotoxin Preconditioning Reprograms S1 Tubules and Macrophages to Protect the Kidney.

    Science.gov (United States)

    Hato, Takashi; Zollman, Amy; Plotkin, Zoya; El-Achkar, Tarek M; Maier, Bernhard F; Pay, S Louise; Dube, Shataakshi; Cabral, Pablo; Yoshimoto, Momoko; McClintick, Jeanette; Dagher, Pierre C

    2018-01-01

    Preconditioning with a low dose of endotoxin confers unparalleled protection against otherwise lethal models of sepsis. The mechanisms of preconditioning have been investigated extensively in isolated immune cells such as macrophages. However, the role of tissue in mediating the protective response generated by preconditioning remains unknown. Here, using the kidney as a model organ, we investigated cell type-specific responses to preconditioning. Compared with preadministration of vehicle, endotoxin preconditioning in the cecal ligation and puncture mouse model of sepsis led to significantly enhanced survival and reduced bacterial load in several organs. Furthermore, endotoxin preconditioning reduced serum levels of proinflammatory cytokines, upregulated molecular pathways involved in phagocytosis, and prevented the renal function decline and injury induced in mice by a toxic dose of endotoxin. The protective phenotype involved the clustering of macrophages around S1 segments of proximal tubules, and full renal protection required both macrophages and renal tubular cells. Using unbiased S1 transcriptomic and tissue metabolomic approaches, we identified multiple protective molecules that were operative in preconditioned animals, including molecules involved in antibacterial defense, redox balance, and tissue healing. We conclude that preconditioning reprograms macrophages and tubules to generate a protective environment, in which tissue health is preserved and immunity is controlled yet effective. Endotoxin preconditioning can thus be used as a discovery platform, and understanding the role and participation of both tissue and macrophages will help refine targeted therapies for sepsis. Copyright © 2018 by the American Society of Nephrology.

  17. Plasma endotoxin activity in Eastern grey kangaroos (Macropus giganteus) with lumpy jaw disease.

    Science.gov (United States)

    Sotohira, Yukari; Suzuki, Kazuyuki; Otsuka, Marina; Tsuchiya, Masakazu; Shimamori, Toshio; Nishi, Yasunobu; Tsukano, Kenji; Asakawa, Mitsuhiko

    2017-06-29

    Progressive pyogranulomatous osteomyelitis involving the mandible or maxilla of captive macropods, referred to as "Lumpy jaw disease (LJD)", is one of the most significant causes of illness and death in captive macropods. The aim of the present study was to evaluate the relationship between the severity of LJD and plasma endotoxin activity in kangaroos. Plasma samples obtained from moderate (n=24) and severe LJD (n=12), and healthy kangaroos (n=46), were diluted 1:20 in endotoxin-free water and heated to 80°C for 10 min. Plasma endotoxin activity was measured using the Limulus amebocyte lysate (LAL)-kinetic turbidimetric (KT) assay. Plasma endotoxin activity was higher in kangaroos with severe LJD (0.199 ± 0.157 EU/ml) than in those with moderate LJD (0.051 ± 0.012 EU/ml, P<0.001) and healthy controls (0.057 ± 0.028 EU/ml, P<0.001). Our results suggest that the severity of LJD in captive macropods may be related to the plasma endotoxin activity.

  18. The efficient removal of endotoxins from insulin using quaternized polyethyleneimine-coated porous zirconia.

    Science.gov (United States)

    McNeff, C; Zhao, Q; Almlöf, E; Flickinger, M; Carr, P W

    1999-10-15

    The synthesis and use of a zirconia-based, alkali-stable strong anion-exchange stationary phase are described for the removal of pyrogenic lipopolysaccharides (LPS) from insulin. The strong anion-exchange material is produced by deposition of polyethyleneimine (PEI) onto porous zirconia particles, followed by cross-linking with a novel reagent, 1,2-bis-(2-iodoethoxy) ethane, and quaternization with iodomethane. Physical characterization of the chromatographic support shows that it has an ion-exchange capacity of 0.6 mmol/g, and 82% of the amine sites on the surface are in quaternized form. Isocratic elution of small benzoic acid derivatives shows good column efficiency. The two primary virtues of this material are its chemical stability under alkali conditions (up to pH 13) and its lower hydrophobicity compared to previously described alkali-stable PEI-coated zirconia supports cross-linked with 1,10-diiododecane. Using this new zirconia-based phase, a purification protocol is developed for the efficient removal of Escherichia coli 0111:B4 LPS from bovine insulin samples. An endotoxin clearance rate of up to 1.3 x 10(8) was attained. Endotoxin levels were reduced to less than 5 endotoxin units/ml even at initial contamination levels as high as 5.0 x 10(6) endotoxin units/ml. Furthermore, endotoxin adsorbed to the porous zirconia column may be easily removed (depyrogenated) using alkali for repeated purification cycles. Copyright 1999 Academic Press.

  19. 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.

  20. Fingolimod against endotoxin-induced fetal brain injury in a rat model.

    Science.gov (United States)

    Yavuz, And; Sezik, Mekin; Ozmen, Ozlem; Asci, Halil

    2017-11-01

    Fingolimod is a sphingosine-1-phosphate receptor modulator used for multiple sclerosis treatment and acts on cellular processes such as apoptosis, endothelial permeability, and inflammation. We hypothesized that fingolimod has a positive effect on alleviating preterm fetal brain injury. Sixteen pregnant rats were divided into four groups of four rats each. On gestational day 17, i.p. endotoxin was injected to induce fetal brain injury, followed by i.p. fingolimod (4 mg/kg maternal weight). Hysterotomy for preterm delivery was performed 6 h after fingolimod. The study groups included (i) vehicle controls (i.p. normal saline only); (ii) positive controls (endotoxin plus saline); (iii) saline plus fingolimod; and (iv) endotoxin plus fingolimod treatment. Brain tissues of the pups were dissected for evaluation of interleukin (IL)-6, caspase-3, and S100β on immunohistochemistry. Maternal fingolimod treatment attenuated endotoxin-related fetal brain injury and led to lower immunoreactions for IL-6, caspase-3, and S100β compared with endotoxin controls (P < 0.0001 for all comparisons). Antenatal maternal fingolimod therapy had fetal neuroprotective effects by alleviating preterm birth-related fetal brain injury with inhibitory effects on inflammation and apoptosis. © 2017 Japan Society of Obstetrics and Gynecology.

  1. 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

    , 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...

  2. Optimization of airborne endotoxin exposure assessment: Effects of filter type, transport conditions, extraction solutions, and storage of samples and extracts

    NARCIS (Netherlands)

    Spaan, S.; Heederik, D.J.J.; Thorne, P.S.; Wouters, I.M.

    2007-01-01

    Endotoxin exposure occurs in homes and occupational environments and is known to cause adverse health effects. In order to compare results from different studies and establish standards, airborne endotoxin exposures should be assessed using standardized methods. Although the European Committee for

  3. A biological study establishing the endotoxin limit for in vitro proliferation of human mesenchymal stem cells

    Directory of Open Access Journals (Sweden)

    Yusuke Nomura

    2017-12-01

    Conclusions: Since endotoxins can affect various cellular functions, an endotoxin limit should be set for in vitro MSC cultures. The lowest observed adverse effect level was determined to be 0.1 ng/ml based on the effect on MSC proliferation.

  4. Actions of four organic acids in radix isatidis on endotoxin-neutralization investigated by kinetic turbidimetric assay.

    Science.gov (United States)

    Ma, Li; He, Ying-jun; Li, You; Gong, Mu-xin

    2012-06-01

    To investigate anti-endotoxin action of four OAs reacted with endotoxin by the LAL assay with KTA. Using a incubating kinetic tube reader and kinetic turbidimetric assay (KTA), the concentration-response time curve of endotoxin reacted with limulus amebocyte lysate (LAL) at 37 degrees C were obtained and the action of four organic acids (OAs) on it were investigated. The four OAs were benzoic acid, salicylic acid, syringic acid and 2-amino-benzoic acid from Radix isatidis. Meanwhile, the temperature variation caused by endotoxin with the four OAs was studied by the rabbit pyrogen test (RPT). It was showed that a low concentration (1 mg/mL) of the four OAs had a little effect of anti-endotoxin, and when the concentrations of the four OAs were 30 mg/mL, the endotoxin was neutralized completely. The relationships between the concentrations of endotoxin and the OAs were all linear with correlation coefficients of greater than 0.9995, indicating that the four OAs all had strong anti-endotoxin action, while syringic acid had the strongest action among the four OAs with IC50 of 12.84 mg/mL. The investigations of KTA agreed well with the results obtained by means of RPT.

  5. 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

    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

  6. 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

  7. 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 sepsi...

  8. A NEW ANIMAL-MODEL FOR HUMAN PREECLAMPSIA - ULTRA-LOW-DOSE ENDOTOXIN INFUSION IN PREGNANT RATS

    NARCIS (Netherlands)

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

    OBJECTIVE: An animal model for preeclampsia was developed by means of an ultra-low-dose endotoxin infusion protocol in conscious pregnant rats. STUDY DESIGN: Rats received a permanent jugular vein cannula on day 0 of pregnancy, through which endotoxin (1.0 mu/kg body weight) (n = 10) or saline

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

    DEFF Research Database (Denmark)

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

    2008-01-01

    The aim of this study was to investigate to what extent single nucleotide polymorphisms (SNPs) in promoter regions of genes of Toll-like receptor (TLR)-4, tumour necrosis factor (TNF)-alpha, interleukin (IL)-18, interferon (IFN)-gamma, IL-6 and IL-10 affect the cytokine response during a controlled......-607, IFN-gamma+874, IL-6-174, IL-10-592 and IL-10-1082) and endotoxin-induced changes in plasma levels of TNF-alpha, IL-6 and IL-10. IL-18 levels were unaffected by endotoxin. In conclusion, the investigated SNPs did not affect endotoxin-induced low-grade cytokine production of TNF-alpha, IL-6, IL-18 or IL......-10 in healthy young men. Previous reports of a major heritability factor in the inflammatory response may be due to other target genes or effects in older age groups or women Udgivelsesdato: 2008/4...

  10. Growth and endotoxin production of Yersinia enterocolitica and Enterobacter agglomerans in packed erythrocytes.

    Science.gov (United States)

    Arduino, M J; Bland, L A; Tipple, M A; Aguero, S M; Favero, M S; Jarvis, W R

    1989-07-01

    Since 1987, the Centers for Disease Control investigated six cases of transfusion-associated sepsis. All six patients developed septic shock after receiving units of packed erythrocytes (PRBCs) contaminated with Yersinia enterocolitica (five patients) and Enterobacter agglomerans (one patient); three of the blood recipients died. We studied the growth and endotoxin production of Y. enterocolitica and E. agglomerans in units of PRBCs stored at 4 degrees C for 60 days. When PRBCs were inoculated with 0.1 to 1.0 CFU of these organisms per ml, both Y. enterocolitica and E. agglomerans entered log-phase growth 2 to 3 weeks after inoculation; generation times were 15 and 22 h, respectively. Endotoxin was first detected at 3 weeks following inoculation, and the concentration paralleled the log phase of growth of the strains tested. These data show that prolonged storage of PRBCs at 4 degrees C provides conditions that allow these two organisms to grow and subsequently produce high concentrations of endotoxin.

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

    DEFF Research Database (Denmark)

    Basinas, Ioannis; Schlünssen, Vivi; Heederik, Dick

    2012-01-01

    E-mediated sensitisation to common allergens and self-reported health symptoms were assessed using logistic regression and generalised additive modelling. Adjustments were made for study, age, sex, atopic predisposition, smoking habit and farm childhood. Heterogeneity was assessed by analysis stratified by study. Results......, and levels above 100 EU/m3 significantly increased the risk of chronic bronchitis (peffect modification except for allergic sensitisation. Only among workers without a farm childhood, endotoxin exposure was inversely associated with allergic sensitisation....... Heterogeneity was primarily present for biofuel workers. Conclusions Occupational endotoxin exposure has a protective effect on allergic sensitisation and hay fever but increases the risk for organic dust toxic syndrome and chronic bronchitis. Endotoxin's protective effects are most clearly observed among...

  12. Panum's studies on "putrid poison" 1856. An early description of endotoxin

    DEFF Research Database (Denmark)

    Kolmos, Hans Jørn

    2006-01-01

    In 1855-1856 the Danish physiologist, Peter Ludvig Panum (1820-1885) performed a series of remarkable experiments on "putrid poison", a hypothetical substance claimed to be responsible for the symptoms and signs seen in patients with sepsis. Dogs were given intravenous infusions of putrefying......, but insoluble in alcohol, and with preserved activity after long-term boiling. "Putrid poison" has striking similarities with endotoxin, a cell wall product of Gram-negative bacteria and a powerful inducer of inflammation and septic shock. Thanks to Panum's carefully arranged experiments and meticulous...... recording of observations it is fair to conclude that "putrid poison" was endotoxin, and as such he deserves credit for being the first to have described endotoxin. Panum published his observations twice, in Danish in 1856, and in German in 1874. At first he rejected the possibility that bacteria could play...

  13. Possible mechanism for preterm labor associated with bacterial infection. I. Stimulation of phosphoinositide metabolism by endotoxin in endometrial fibroblasts

    International Nuclear Information System (INIS)

    Khan, A.A.; Imai, A.; Tamaya, T.

    1990-01-01

    Growing evidence suggests an association between intra-amniotic infection and premature initiation of parturition. We recently demonstrated that some factor(s) including endotoxin produced by the organism stimulates endogenous phospholipase A2 resulting in liberation of arachidonic acid and prostaglandin formation. The studies presented in this report were designated to evaluate the mechanism for endotoxin to stimulate phospholipase A2 using human endometrial fibroblasts. Exposure of the fibroblasts to endotoxin from Escherichia coli in the presence of [ 32 P] phosphate increased 32 P-labeling of phosphatidic acid (PA) and phosphatidyl-inositol (PI) in a dose-dependent and a time-dependent manners. The PA labeling occurred without a measurable lag time. These findings demonstrate that the endotoxin stimulates phosphoinositide metabolism in human endometrial fibroblasts by a receptor-mediated mechanism. Membrane phosphoinositide turnover stimulated by endotoxin results in cytosolic Ca 2+ increment, liberation of arachidonic acid, which may be involved in the initiation of parturition

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

    Science.gov (United States)

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

    2016-02-01

    This study evaluated the variation over a year in airborne endotoxin levels in the indoor environment of five university laboratories in Seoul, South Korea, and examined the micro-environmental factors that influenced endotoxin levels. These included temperature, relative humidity, CO2, CO, illumination, and wind velocity. A total of 174 air samples were collected and analyzed using the kinetic limulus amebocyte lysate assay. Endotoxin levels ranged from analysis showed that the environmental factors affecting endotoxin levels were temperature (coefficient=-0.388, p<0.001) and illumination (coefficient=-0.370, p<0.001). Strategies aimed at reducing airborne endotoxin levels in the indoor environments may be most effective if they focus on illumination. Copyright © 2015 Elsevier Inc. All rights reserved.

  15. Endotoxin exposure in sewage treatment workers: investigation of exposure variability and comparison of analytical techniques.

    Science.gov (United States)

    Spaan, Suzanne; Smit, Lidwien A M; Eduard, Wijnand; Larsson, Lennart; Arts, Huib J J M; Wouters, Inge M; Heederik, Dick J J

    2008-01-01

    Objectives were to give an overview of endotoxin exposure and its determinants in sewage treatment workers, and to study exposure to culturable and non-culturable microorganisms and the applicability of the LAL assay in this work environment. In 43 Dutch sewage treatment plants 470 full-shift, 123 task-based personal and 54 stationary inhalable dust samples were collected. Endotoxin concentration was determined with the LAL-assay. Mixed effects models were used to investigate possible determinants of exposure. Simultaneous parallel filter samples, impinger samples and viable total bacteria and Gram-negative bacterial samples were taken to compare analytical techniques. Filter and impinger samples were analyzed with the LAL-assay, gas chromatography-mass spectrometry (GC-MS) and fluorescence microscopy. Endotoxin exposure levels were moderate to low (geometric mean personal exposure 27 EU/m(3), stationary 33 EU/m(3), task-based 64 EU/m(3)), yet differences between jobs and sources and some determinants of exposure were identified. Exposure varied more from day to day than between workers. Concentrations in filter samples were higher and more consistent than in impinger samples. Fungi and Gram-positive bacteria were found in higher levels than Gram-negative bacteria. The LAL assay and GC-MS showed comparable endotoxin levels. Endotoxin exposure in Dutch sewage treatment workers was relatively low. Comparison of sampling and analytical techniques suggests that the LAL-assay did not result in much exposure misclassification. It thus seems justified to perform filter measurements in combination with the LAL-assay to measure endotoxin exposure in sewage treatment plants.

  16. House Dust Endotoxin and Peripheral Leukocyte Counts: Results from Two Large Epidemiologic Studies.

    Science.gov (United States)

    Fessler, Michael B; Carnes, Megan U; Salo, Päivi M; Wilkerson, Jesse; Cohn, Richard D; King, Debra; Hoppin, Jane A; Sandler, Dale P; Travlos, Greg; London, Stephanie; Thorne, Peter; Zeldin, Darryl

    2017-05-31

    The peripheral leukocyte count is a biomarker of inflammation and is associated with human all-cause mortality. Although causes of acute leukocytosis are well-described, chronic environmental determinants of leukocyte number are less well understood. We investigated the relationship between house dust endotoxin concentration and peripheral leukocyte counts in human subjects. The endotoxin–leukocyte relationship was evaluated by linear regression in the National Health and Nutrition Examination Survey (NHANES) 2005–2006 (n=6,254) and the Agricultural Lung Health Study (ALHS; n=1,708). In the ALHS, we tested for a gene [Toll-like Receptor 4 ( TLR4 ), encoding the endotoxin receptor]-by-environment interaction in the endotoxin–leukocyte relationship using regression models with an interaction term. There is a statistically significant, positive association between endotoxin concentration and total leukocyte number [estimated change, 0.186×10 3 /μL (95% CI: 0.070, 0.301×10 3 /μL) per 10-fold change in endotoxin; p=0.004) in the NHANES. Similar positive associations were found for monocytes, lymphocytes, and neutrophils. Stratified analyses revealed possible effect modification by asthma and chronic obstructive pulmonary disease. We observed similar associations in the ALHS. For total leukocytes, there was suggestive evidence in the ALHS of a gene-by-environment interaction for minor allele carrier status at the TLR4 haplotype defined by rs4986790 and rs4986791 (interaction p=0.15). This is, to our knowledge, the first report of an association between house dust endotoxin and leukocyte count in a national survey. The finding was replicated in a farming population. Peripheral leukocyte count may be influenced by residential endotoxin exposure in diverse settings. https://doi.org/10.1289/EHP661.

  17. Endotoxin-induced inflammation disturbs melatonin secretion in ewe

    Directory of Open Access Journals (Sweden)

    Andrzej Przemysław Herman

    2017-12-01

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

  18. Intolerant tolerance.

    Science.gov (United States)

    Khushf, G

    1994-04-01

    The Hyde Amendment and Roman Catholic attempts to put restrictions on Title X funding have been criticized for being intolerant. However, such criticism fails to appreciate that there are two competing notions of tolerance, one focusing on the limits of state force and accepting pluralism as unavoidable, and the other focusing on the limits of knowledge and advancing pluralism as a good. These two types of tolerance, illustrated in the writings of John Locke and J.S. Mill, each involve an intolerance. In a pluralistic context where the free exercise of religion is respected, John Locke's account of tolerance is preferable. However, it (in a reconstructed form) leads to a minimal state. Positive entitlements to benefits like artificial contraception or nontherapeutic abortions can legitimately be resisted, because an intolerance has already been shown with respect to those that consider the benefit immoral, since their resources have been coopted by taxation to advance an end that is contrary to their own. There is a sliding scale from tolerance (viewed as forbearance) to the affirmation of communal integrity, and this scale maps on to the continuum from negative to positive rights.

  19. Variation in the ovine cortisol response to systemic bacterial endotoxin challenge is predominantly determined by signalling within the hypothalamic-pituitary-adrenal axis

    International Nuclear Information System (INIS)

    You Qiumei; Karrow, Niel A.; Cao Honghe; Rodriguez, Alexander; Mallard, Bonnie A.; Boermans, Herman J.

    2008-01-01

    Bi-directional communication between the neuroendocrine and immune systems is designed, in part, to maintain or restore homeostasis during physiological stress. Exposure to endotoxin during Gram-negative bacterial infection for example, elicits the release of pro-inflammatory cytokines that activate the hypothalamic-pituitary-adrenal axis (HPAA). The secretion of adrenal glucocorticoids subsequently down regulates the host inflammatory response, minimizing potential tissue damage. Sequence and epigenetic variants in genes involved in regulating the neuroendocrine and immune systems are likely to contribute to individual differences in the HPAA response, and this may influence the host anti-inflammatory response to toxin exposure and susceptibility to inflammatory disease. In this study, high (HCR) and low (LCR) cortisol responders were selected from a normal population of 110 female sheep challenged iv with Escherichia coli endotoxin (400 ng/kg) to identify potential determinants that contribute to variation in the cortisol response phenotype. This phenotype was stable over several years in the HCR and LCR animals, and did not appear to be attributed to differences in expression of hepatic immune-related genes or systemic pro-inflammatory cytokine concentrations. Mechanistic studies using corticotrophin-releasing factor (0.5 μg/kg body weight), arginine vasopressin (0.5 μg/kg), and adrenocorticotropic hormone (0.5 μg/kg) administered iv demonstrated that variation in this phenotype is largely determined by signalling within the HPAA. Future studies will use this ovine HCR/LCR model to investigate potential genetic and epigenetic variants that may contribute to variation in cortisol responsiveness to bacterial endotoxin

  20. Effect of the selective NMDA NR2B antagonist, ifenprodil, on acute tolerance to ethanol-induced motor impairment in adolescent and adult rats.

    Science.gov (United States)

    Ramirez, Ruby Liane; Varlinskaya, Elena I; Spear, Linda P

    2011-06-01

    Adolescent rats have been observed to be less sensitive than adults to a number of acute ethanol effects, including ethanol-induced motor impairment. These adolescent insensitivities may be related in part to the more rapid emergence of within session (acute) tolerance in adolescents than adults. Adolescent-related alterations in neural systems that serve as ethanol target sites, including changes in NMDA receptor subunit expression, may influence the responsiveness of adolescents to acute ethanol effects. This study explored the role of NMDA NR2B receptors in the development of acute tolerance to ethanol-induced motor impairment in male adolescent [postnatal day (P)28-30] and adult (P68-70) Sprague-Dawley rats. Motor-impairing effects of ethanol on the stationary inclined plane and blood ethanol concentrations (BECs) were examined following challenge at each age with a functionally equivalent ethanol dose (adolescents: 2.25 g/kg; adults: 1.5 g/kg). Data were collected at two postinjection intervals (10 or 60 minutes) to compare rate of recovery from ethanol intoxication with BEC declines using the Radlow approach (Radlow, 1994) and changes in motor impairment/BEC ratios over time for assessing acute tolerance. Both vehicle-treated adolescent and adult animals showed similar acute tolerance development to the motor-impairing effects of ethanol at these functionally equivalent doses on the stationary inclined plane, as indexed by an increasing time-dependent dissociation between BECs and ethanol-induced motor impairment, with motor impairment declining faster than BECs, as well as by significant declines in motor impairment/BEC ratios over time. Acute tolerance development was reliably blocked by administration of the NR2B antagonist, ifenprodil, (5.0 mg/kg), in adult rats, whereas adolescents were affected by a higher dose (10.0 mg/kg). These data support the suggestion that alterations in NMDA receptor systems occurring during adolescence may contribute to

  1. Quantification of cultivable bacteria and endotoxin in post-treatment apical periodontitis before and after chemo-mechanical preparation.

    Science.gov (United States)

    Endo, M S; Martinho, F C; Zaia, A A; Ferraz, C C R; Almeida, J F A; Gomes, B P F A

    2012-10-01

    This clinical study was conducted to quantify cultivable bacteria and endotoxin in root canals with post-treatment apical periodontitis by correlating their levels with clinical features and to evaluate the effect of chemo-mechanical preparation (CMP) with 2 % chlorhexidine gel + 17 % EDTA on bacterial and endotoxin removal/elimination. Moreover, target strict Gram-negative anaerobic bacteria were investigated by polymerase chain reaction (PCR). Fifteen teeth with post-treatment apical periodontitis were sampled before (s1) and after (s2) CMP. Culture techniques determined the number of colony-forming units (CFU). PCR (16S rDNA) and limulus amebocyte lysate (LAL) assay were used for bacterial and endotoxin detection, respectively. Prevotella nigrescens (4/15), Prevotella intermedia (2/15), and Tannerella forsythia (2/15) were the most frequently detected species. Endotoxin was recovered in 100 % of the samples. At s1, bacteria and endotoxin were detected at a median value of 5.14 × 10(3) CFU/mL and 3.96 EU/mL, respectively. Higher levels of endotoxin were related to a larger size of radiolucent area (>5 mm) (p area in the periapical region. Moreover, CMP was effective in reducing both bacterial and endotoxin contents in post-treatment apical periodontitis.

  2. 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, Pkangaroos 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.

  3. Endotoxin pretreatment modifies peristalsis and attenuates the antipropulsive action of adrenoceptor agonists in the guinea-pig small intestine.

    Science.gov (United States)

    Fruhwald, S; Herk, E; Schöll, G; Shahbazian, A; Hammer, H F; Metzler, H; Holzer, P

    2004-04-01

    The action of endotoxin to alter gastrointestinal motility in vivo may reflect a direct effect on the gut or result from vascular and other systemic manifestations of this sepsis model. Here we examined whether in vivo pretreatment of guinea-pigs with endotoxin modifies peristalsis in the isolated gut and influences the antipropulsive action of adrenoceptor agonists. Distension-induced peristalsis was recorded in fluid-perfused segments of the small intestine taken from animals pretreated intraperitoneally with endotoxin (1 mg kg(-1)Escherichia coli lipopolysaccharide) or vehicle 4 or 20 h before. Clonidine, adrenaline, noradrenaline, dopamine and dobutamine inhibited peristalsis with differential potency. Endotoxin pretreatment lowered the peristaltic pressure threshold and altered other parameters of baseline peristalsis in a time-related manner. The potency and efficacy of clonidine to inhibit peristalsis were markedly decreased after endotoxin administration, while the potency of the other test drugs was less attenuated. The antipropulsive action of clonidine in control segments was reduced by yohimbine and prazosin, whereas in segments from endotoxin-pretreated animals it was antagonized by yohimbine but not prazosin. We conclude that systemic endotoxin pretreatment of guinea-pigs modifies baseline peristalsis by an action on the gut and inhibits the antipropulsive action of adrenoceptor agonists through changes in adrenoceptor activity.

  4. 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

  5. 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.

    Science.gov (United States)

    Soncin, Sabrina; Lo Cicero, Viviana; Astori, Giuseppe; Soldati, Gianni; Gola, Mauro; Sürder, Daniel; Moccetti, Tiziano

    2009-09-08

    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. 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. The calculated MVD and endotoxin limit were 780x 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). 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 ensuring the safety and efficacy of the next generation of ATMPs. Personnel

  6. The opiate sufentanil alters the inflammatory, endocrine, and metabolic responses to endotoxin in dogs

    NARCIS (Netherlands)

    Moeniralam, H. S.; Endert, E.; Ackermans, M. T.; van Lanschot, J. J.; Sauerwein, H. P.; Romijn, J. A.

    1998-01-01

    Sufentanil is a synthetic mu-opioid receptor agonist frequently used in anesthesia and critically ill patients. To evaluate the effects of sufentanil on the inflammatory, neuroendocrine, and metabolic responses to endotoxin, we studied six dogs during saline infusion (control), during sufentanil

  7. Efficacy of an extracorporeal endotoxin adsorber system during hyperdynamic porcine endotoxemia.

    Science.gov (United States)

    Bracht, H; Hauser, B; Ivanyi, Z; Asfar, P; Ehrmann, U; Brueckner, U B; Georgieff, M; Radermacher, P; Buttenschon, K

    2009-01-01

    Endotoxemia is a crucial factor in the pathogenesis of sepsis. Elimination of endotoxin is aimed at the reduction of sepsis-related morbidity and lethality. The objective of this study was to examine the impact of an endotoxin adsorber on hemodynamics, O(2) exchange and metabolism during resuscitated porcine endotoxemia. Twenty pigs were randomized into 2 intervention groups (n = 7 each) and 1 control group (n = 6). Endotoxemia was induced by continuous intravenous application of lipopolysaccharide for 8 h. Adsorber therapy was started at the same time as the induction of endotoxemia or 2 h later. An extracorporeal hemoperfusion device using immobilized human serum albumin for endotoxin adsorption was used. Hemodynamic, metabolic and acid-base parameters, as well as the kinetics of interleukin (IL)-6, IL-8, IL-10 and tumor necrosis factor-alpha, were characteristic for endotoxic shock. Endotoxin plasma levels were low (arterial, hepatic and portal vein). None of the parameters were significantly influenced by the adsorber system. Despite typical clinical signs of endotoxemia, the adsorber system had no significant effect on hemodynamic, metabolic and acid-base parameters during endotoxic shock. The reasons for the absence of an effect are elusive; however, failure of the method per se or exceeded capacity of the adsorber cannot be excluded. Copyright (c) 2009 S. Karger AG, Basel.

  8. Pharmacological modulation of the endotoxin-induced increase in plasminogen activator inhibitor activity in rats

    NARCIS (Netherlands)

    Emeis, J.J.; Hoogen, C.M. van den

    1992-01-01

    Pharmacological modulation of the in vivo induction of plasminogen activator inhibitor type-1 (PAI-1) synthesis was studied in rats using the induction of PAI-1 by endotoxin as a model system. Both the cyclooxygenase inhibitors acetylsalicylic acid and indomethacin enhanced PAI-1 induction. The

  9. Differential effects of air conditioning type on residential endotoxin levels in a semi-arid climate.

    Science.gov (United States)

    Johnston, J D; Kruman, B A; Nelson, M C; Merrill, R M; Graul, R J; Hoybjerg, T G; Tuttle, S C; Myers, S J; Cook, R B; Weber, K S

    2017-09-01

    Residential endotoxin exposure is associated with protective and pathogenic health outcomes. Evaporative coolers, an energy-efficient type of air conditioner used in dry climates, are a potential source of indoor endotoxins; however, this association is largely unstudied. We collected settled dust biannually from four locations in homes with evaporative coolers (n=18) and central air conditioners (n=22) in Utah County, Utah (USA), during winter (Jan-Apr) and summer (Aug-Sept), 2014. Dust samples (n=281) were analyzed by the Limulus amebocyte lysate test. Housing factors were measured by survey, and indoor temperature and relative humidity measures were collected during both seasons. Endotoxin concentrations (EU/mg) were significantly higher in homes with evaporative coolers from mattress and bedroom floor samples during both seasons. Endotoxin surface loads (EU/m 2 ) were significantly higher in homes with evaporative coolers from mattress and bedroom floor samples during both seasons and in upholstered furniture during winter. For the nine significant season-by-location comparisons, EU/mg and EU/m 2 were approximately three to six times greater in homes using evaporative coolers. A plausible explanation for these findings is that evaporative coolers serve as a reservoir and distribution system for Gram-negative bacteria or their cell wall components in homes. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  10. Determination of the plasmid size and location of d-endotoxin genes ...

    African Journals Online (AJOL)

    The genes encoding the d-endotoxins of Bacillus thuringiensis are located on plasmids ranging in size from 45 to 1000 kb. Plasmid size and variety are diagnostic features for characterizing subspecies of this aerobic spore-forming crystalliferous entomopathogen. Two of 25 B. thuringiensis isolates obtained from Middle ...

  11. Effect of enrofloxacin treatment on plasma endotoxin during bovine Escherichia coli mastitis

    NARCIS (Netherlands)

    Dosogne, H.; Meyer, E.; Sturk, A.; van Loon, J.; Massart-Leën, A. M.; Burvenich, C.

    2002-01-01

    OBJECTIVE AND DESIGN: To investigate the effect of enrofloxacin on endotoxin resorption during bovine Escherichia coli mastitis. ANIMALS: 12 healthy early post partum Holstein cows. TREATMENT: Mastitis was induced by intramammary infusion of 10(4) cfu E. coli P4:032. Six cows were treated twice

  12. Development of an in vitro Endotoxin Test for Monoolein–Water ...

    African Journals Online (AJOL)

    Purpose: Drugs that are administered by parenteral route must be apyrogenic. The aim of this study was to develop an in vitro endotoxin test for liquid crystalline gels for use as implants, using a monoolein–water liquid crystalline gel as a model. Methods: The gel-clot technique was used. The gel was dissolved first in ...

  13. Endotoxin-induced liver damage in rats is minimized by beta 2-adrenoceptor stimulation.

    NARCIS (Netherlands)

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

    2004-01-01

    OBJECTIVE AND DESIGN: To investigate the effects of beta(2)-adrenoceptor (beta(2)-AR) stimulation on endotoxin-induced liver damage and systemic cytokine levels in rats. SUBJECTS: Standard male Wistar rats. TREATMENT: A disease-model of lipopolysaccharide (LPS)-induced acute systemic inflammation

  14. 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

  15. Modulation of reactive oxygen species production alters responses of lung inflammatory cells to endotoxin

    Czech Academy of Sciences Publication Activity Database

    Krejčová, Daniela; Konopka, Roman; Lojek, Antonín; Číž, Milan; Kubala, Lukáš

    2006-01-01

    Roč. 40, č. 1 (2006), S112-S112 ISSN 1071-5762 R&D Projects: GA ČR(CZ) GA524/06/1197 Institutional research plan: CEZ:AV0Z50040507 Keywords : myeloperoxidase * inflammation * endotoxin Subject RIV: BO - Biophysics

  16. 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

  17. Endotoxin levels in settled airborne dust in European schools : The HITEA school study

    NARCIS (Netherlands)

    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

    2014-01-01

    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

  18. 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...

  19. Development of an in vitro Endotoxin Test for Monoolein–Water ...

    African Journals Online (AJOL)

    Erah

    Loaded Monoolein Gel Intended to Treat. Chronic Osteomyelitis. J. Pharmacol. Toxicol.,. 2008; 3(5): 386-393. 13 Magalahaes PO, Lopes AM, Mazolla PG, Rangel-. Yagui C, Penna TCV, Pessoa Jr A. Methods of. Endotoxins. Removal from. Biological. Preparations: a Review. J Pharm Pharmaceut. Sci. 2007; 10(3): 388-404.

  20. Effects of Bacillus thuringiensis CRY1A(c) δ-endotoxin on growth ...

    African Journals Online (AJOL)

    STORAGESEVER

    2010-01-04

    Jan 4, 2010 ... The recent introduction of Bt maize and Bt cotton transgenic crops into Africa has raised concerns on their potential short and long-term ecological effects on the environment. The effects of Bacillus thuringiensis (Bt) Cry1A(c) δ-endotoxin on the growth, nodulation and productivity of two leguminous.

  1. Effects of Bacillus thuringiensis CRY1A(c) d-endotoxin on growth ...

    African Journals Online (AJOL)

    The recent introduction of Bt maize and Bt cotton transgenic crops into Africa has raised concerns on their potential short and long-term ecological effects on the environment. The effects of Bacillus thuringiensis (Bt) Cry1A(c) d-endotoxin on the growth, nodulation and productivity of two leguminous plants grown in clay soil ...

  2. Endotoxin-induced uveitis in the rat. The significance of intraocular interleukin-6

    NARCIS (Netherlands)

    Hoekzema, R.; Verhagen, C.; van Haren, M.; Kijlstra, A.

    1992-01-01

    The potential role of interleukin-6 (IL-6) was studied as an inflammatory mediator of endotoxin (or lipopolysaccharide [LPS])-induced uveitis (EIU) in the rat. In young Lewis rats, levels of intraocular IL-6, but not serum IL-6, correlated with the severity of uveitis and with aqueous humor protein

  3. Endotoxin induces fibrosis in vascular endothelial cells through a mechanism dependent on transient receptor protein melastatin 7 activity.

    Directory of Open Access Journals (Sweden)

    Cesar Echeverría

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

  4. 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

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

    Science.gov (United States)

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

    2018-03-01

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

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

    Directory of Open Access Journals (Sweden)

    Hussein Ahmed

    2004-01-01

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

  7. The F130S point mutation in the Arabidopsis high-affinity K+ transporter AtHAK5 increases K+ over Na+ and Cs+ selectivity and confers Na+ and Cs+ tolerance to yeast under heterologous expression

    Directory of Open Access Journals (Sweden)

    Fernando eAleman

    2014-09-01

    Full Text Available Potassium (K+ is an essential macronutrient required for plant growth, development and high yield production of crops. Members of group I of the KT/HAK/KUP family of transporters, such as HAK5, are key components for K+ acquisition by plant roots at low external K+ concentrations. Certain abiotic stress conditions such as salinity or Cs+-polluted soils may jeopardize plant K+ nutrition because HAK5-mediated K+ transport is inhibited by Na+ and Cs+. Here, by screening in yeast a randomly-mutated collection of AtHAK5 transporters, a new mutation in AtHAK5 sequence is identified that greatly increases Na+ tolerance. The single point mutation F130S, affecting an amino acid residue conserved in HAK5 transporters from several species, confers high salt tolerance, as well as Cs+ tolerance. This mutation increases more than 100-fold the affinity of AtHAK5 for K+ and reduces the Ki values for Na+ and Cs+, suggesting that the F130 residue may contribute to the structure of the pore region involved in K+ binding. In addition, this mutation increases the Vmax for K+. All this changes occur without increasing the amount of the AtHAK5 protein in yeast and support the idea that this residue is contributing to shape the selectivity filter of the AtHAK5 transporter.

  8. Drought tolerant wheat varieties developed through mutation ...

    African Journals Online (AJOL)

    In search for higher yielding drought tolerant wheat varieties, one of the Kenyan high yielding variety 'Pasa' was irradiated with gamma rays (at 150, 200, and 250gy) in 1997 so as to induce variability and select for drought tolerance. Six mutants ((KM10, KM14, KM15, KM18, KM20 and KM21) were selected at M4 for their ...

  9. Mosapride, a selective serotonin 5-HT4 receptor agonist, and alogliptin, a selective dipeptidyl peptidase-4 inhibitor, exert synergic effects on plasma active GLP-1 levels and glucose tolerance in mice.

    Science.gov (United States)

    Nonogaki, Katsunori; Kaji, Takao

    2015-12-01

    Pharmacologic stimulation of serotonin 5-HT4 receptors increased plasma active glucagon-like-peptide-1 (GLP-1) levels independent of feeding, and that pharmacologic stimulation of 5-HT4 receptors and pharmacologic inhibition of dipeptidyl peptidase-4 exerted synergic effects on plasma active GLP-1 levels and glucose tolerance in mice. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  10. Allocating application to group of consecutive processors in fault-tolerant deadlock-free routing path defined by routers obeying same rules for path selection

    Science.gov (United States)

    Leung, Vitus J [Albuquerque, NM; Phillips, Cynthia A [Albuquerque, NM; Bender, Michael A [East Northport, NY; Bunde, David P [Urbana, IL

    2009-07-21

    In a multiple processor computing apparatus, directional routing restrictions and a logical channel construct permit fault tolerant, deadlock-free routing. Processor allocation can be performed by creating a linear ordering of the processors based on routing rules used for routing communications between the processors. The linear ordering can assume a loop configuration, and bin-packing is applied to this loop configuration. The interconnection of the processors can be conceptualized as a generally rectangular 3-dimensional grid, and the MC allocation algorithm is applied with respect to the 3-dimensional grid.

  11. Infectious Tolerance

    OpenAIRE

    Jonuleit, Helmut; Schmitt, Edgar; Kakirman, Hacer; Stassen, Michael; Knop, Jürgen; Enk, Alexander H.

    2002-01-01

    Regulatory CD4+CD25+ T cells (Treg) are mandatory for maintaining immunologic self-tolerance. We demonstrate that the cell-cell contact–mediated suppression of conventional CD4+ T cells by human CD25+ Treg cells is fixation resistant, independent from membrane-bound TGF-β but requires activation and protein synthesis of CD25+ Treg cells. Coactivation of CD25+ Treg cells with Treg cell–depleted CD4+ T cells results in anergized CD4+ T cells that in turn inhibit the activation of conventional, ...

  12. Infectious Tolerance

    Science.gov (United States)

    Jonuleit, Helmut; Schmitt, Edgar; Kakirman, Hacer; Stassen, Michael; Knop, Jürgen; Enk, Alexander H.

    2002-01-01

    Regulatory CD4+CD25+ T cells (Treg) are mandatory for maintaining immunologic self-tolerance. We demonstrate that the cell-cell contact–mediated suppression of conventional CD4+ T cells by human CD25+ Treg cells is fixation resistant, independent from membrane-bound TGF-β but requires activation and protein synthesis of CD25+ Treg cells. Coactivation of CD25+ Treg cells with Treg cell–depleted CD4+ T cells results in anergized CD4+ T cells that in turn inhibit the activation of conventional, freshly isolated CD4+ T helper (Th) cells. This infectious suppressive activity, transferred from CD25+ Treg cells via cell contact, is cell contact–independent and partially mediated by soluble transforming growth factor (TGF)-β. The induction of suppressive properties in conventional CD4+ Th cells represents a mechanism underlying the phenomenon of infectious tolerance. This explains previously published conflicting data on the role of TGF-β in CD25+ Treg cell–induced immunosuppression. PMID:12119350

  13. 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.

  14. Is the association between pet ownership and indoor endotoxin levels confounded or modified by outdoor residential greenspace?

    Science.gov (United States)

    Fuertes, Elaine; Standl, Marie; Markevych, Iana; Bischof, Wolfgang; Heinrich, Joachim

    2018-06-01

    Pet ownership is consistently associated with higher indoor endotoxin concentrations, but may also be related to the amount of greenspace around the home. This study examined whether the association between pet ownership and higher indoor endotoxin concentrations is confounded or modified by residential greenspace. Information on pet ownership was collected at the time of recruitment of the German LISA birth cohort. Endotoxin levels were measured in settled house dust sampled from mothers' mattresses (N=1197) and living room floors (N=390). Greenspace around the home was assessed as the mean Normalized Difference Vegetation Index in 100m, 500m and 1000m buffers around the home, and as the distance to the nearest urban and natural green space. Linear regression models assessed cross-sectional associations between pet ownership and log-transformed endotoxin levels, adjusted for known predictors of endotoxin levels. Confounding by greenspace was assessed by additionally adjusting the models for each greenspace variable. Effect modification was assessed by including interaction terms between pet ownership and each greenspace variable, and by model stratification. Dog and cat ownership were associated with higher endotoxin levels in mothers' mattresses, whereas only dog ownership was associated with endotoxin levels in the floor samples. All associations were highly robust to further adjustment for greenspace, and there was little evidence to suggest any effect modification (interaction terms had p-values >0.05). Residential greenspace did not confound or modify the association between pet ownership and indoor endotoxin levels. Studies should continue investigating whether pets influence the indoor environment only by their presence, or also by acting as transmission vectors of the outdoors. Copyright © 2018 Elsevier B.V. All rights reserved.

  15. Longitudinal distribution of pulmonary vascular resistance after endotoxin administration in sheep.

    Science.gov (United States)

    Pearl, R G; Baer, E R; Siegel, L C; Benson, G V; Rice, S A

    1992-01-01

    Pulmonary hypertension may increase pulmonary capillary pressure and exacerbate pulmonary edema in acute respiratory failure. The effects of pulmonary hypertension on pulmonary capillary pressure depend on the longitudinal distribution of pulmonary vascular resistance. Since pulmonary hypertension occurs during acute respiratory failure, we hypothesized that acute respiratory failure may produce time-dependent changes in the longitudinal distribution of pulmonary vascular resistance. Therefore, we measured pulmonary capillary pressure and the longitudinal distribution of pulmonary vascular resistance in an animal model of acute respiratory failure. Escherichia coli endotoxin (2.5 to 5.0 micrograms/kg) was administered over a 1-hr period in eight anesthetized sheep. Pulmonary and systemic hemodynamics, including pulmonary artery occlusion pressure (PAOP), pulmonary capillary pressure, and the longitudinal distribution of pulmonary vascular resistance, were measured over the next 5 hrs. Pulmonary capillary pressure was estimated by analysis of the pressure decay following pulmonary artery balloon inflation. Endotoxin administration resulted in sustained pulmonary hypertension for the subsequent 5 hrs of the study. Pulmonary capillary pressure was increased 7 mm Hg above baseline at 0.5 and 0.75 hrs during the infusion of endotoxin but returned to baseline values at 1.5 hrs. Despite sustained pulmonary hypertension, pulmonary capillary pressure remained at baseline values for the duration of the study. Similar to pulmonary capillary pressure, pulmonary venous (or postcapillary) resistance was increased approximately four-fold over baseline at 0.5 and 0.75 hrs after initiating endotoxin administration, but returned to baseline values by the end of endotoxin administration and remained at baseline values throughout the remainder of the study. In contrast, pulmonary arterial (or precapillary) resistance remained at values approximately three times baseline during the

  16. Workplace Determinants of Endotoxin Exposure in Dental Healthcare Facilities in South Africa

    Science.gov (United States)

    Singh, Tanusha S.; Bello, Braimoh; Mabe, Onnicah D.; Renton, Kevin; Jeebhay, Mohamed F.

    2010-01-01

    Objectives: Aerosols generated during dental procedures have been reported to contain endotoxin as a result of bacterial contamination of dental unit water lines. This study investigated the determinants of airborne endotoxin exposure in dental healthcare settings. Methods: The study population included dental personnel (n = 454) from five academic dental institutions in South Africa. Personal air samples (n = 413) in various dental jobs and water samples (n = 403) from dental handpieces and basin taps were collected. The chromogenic-1000 limulus amebocyte lysate assay was used to determine endotoxin levels. Exposure metrics were developed on the basis of individually measured exposures and average levels within each job category. Analysis of variance and multivariate linear regression models were constructed to ascertain the determinants of exposure in the dental group. Results: There was a 2-fold variation in personal airborne endotoxin from the least exposed (administration) to the most exposed (laboratory) jobs (geometric mean levels: 2.38 versus 5.63 EU m−3). Three percent of personal samples were above DECOS recommended exposure limit (50 EU m−3). In the univariate linear models, the age of the dental units explained the most variability observed in the personal air samples (R2 = 0.20, P < 0.001), followed by the season of the year (R2 = 0.11, P < 0.001). Other variables such as institution and total number of dental units per institution also explained a modest degree of variability. A multivariate model explaining the greatest variability (adjusted R2 = 0.40, P < 0.001) included: the age of institution buildings, total number of dental units per institution, ambient temperature, ambient air velocity, endotoxin levels in water, job category (staff versus students), dental unit model type and age of dental unit. Conclusions: Apart from job type, dental unit characteristics are important predictors of airborne endotoxin

  17. Assessing the removal potential of soil-aquifer treatment system (soil column) for endotoxin.

    Science.gov (United States)

    Guizani, Mokhtar; Kato, Hideaki; Funamizu, Naoyuki

    2011-06-01

    Soil-aquifer treatment (SAT) of wastewater is an increasingly valued practice for replenishing aquifers due to ease of operation and low maintenance needs and therefore low cost. In this study, we investigated the fate of endotoxins through laboratory-scale SAT soil columns over a four month period. The effluent of rapid sand filtration was run through the columns under gravity flow conditions. Four SAT columns were packed with four different filter materials (fine sand, medium sand, coarse sand and very coarse sand). The effluent of rapid sand filtration (average dissolved organic carbon (DOC) = 4 mg l(-1) and average endotoxin concentration = 4 EU ml(-1)) was collected from a domestic wastewater treatment plant in Sapporo, Japan. DOC removal ranged from 12.5% to greater than 22.5% during the study, with DOC levels averaging 3.1 and 3.5 mg l(-1) for the SAT columns packed with different soils. Endotoxin transformation exhibited different profiles, depending on the time and soil type. Reduction in endotoxin concentration averaged 64.3% and was as high as 86.7% across the soil columns 1, 2, 3 and 4, respectively. While DOC removal was gradual, the reductions in endotoxin levels were rather rapid and most of the removal was achieved in the top layers. Soil with a larger grain size had lower efficiency in removing endotoxin. Tests were performed to evaluate the transformation of organic matter showing endotoxicity and to determine the mechanisms responsible for changes in the structural and size properties of dissolved organic matter (OM) during SAT. Dissolved OM was fractionated using Sep-Pack C18 Cartridges into hydrophobic and hydrophilic fractions. Dialysis tubes with different molecular weight cut-offs were used to perform size fractions of OM showing endotoxicity. Evaluation of the transformation of organic matter showing endotoxicity during SAT indicated that both hydrophobic and large molecules were reduced. Moreover, experimental findings showed that

  18. Respiratory symptoms, fractional exhaled nitric oxide & endotoxin exposure among female flower farm workers in Ethiopia.

    Science.gov (United States)

    Nigatu, Amare W; Bråtveit, Magne; Deressa, Wakgari; Moen, Bente E

    2015-01-01

    Greenhouse workers are exposed to organic dusts, and they are thereby at risk of developing airway disorders. This study aims to measure personal endotoxin exposure, assess respiratory symptoms and measure fractional exhaled nitric oxide (FeNO) among female flower farm workers in Ethiopia. A cross-sectional study involving female workers (n = 248) from four flower farms was conducted. The workers were interviewed for respiratory symptoms using a standard questionnaire. Workers from two of these farms also participated in personal endotoxin sampling (46 workers, 75 measurements) on glass fiber filters (0.2 μm pore size) inside conductive 25 mm Millipore cassettes for sampling of the "total dust" fraction. They also participated in FeNO (n = 114) measurements with a portable electrochemistry-based sensor. Chi-square and independent t-tests were used to compare categorical and continuous variables respectively. A mixed-effects model was used to analyze exposure determinants. Endotoxin exposure had a geometric mean (GM) of 22.8 endotoxin units (EU)/m(3) with a maximum of 180 EU/m(3). Greenhouse workers had significantly higher endotoxin exposure than workers outside the greenhouses (GM = 26.7 vs. 19.3 EU/m(3) respectively; p farm was 21 months. Greenhouse workers had higher prevalence of self-reported respiratory symptoms than those outside greenhouses. However, after adjusting for education only blocked nose remained significant. The FeNO concentration ranged 5-166 ppb (GM = 14 ppb). Two workers had FeNO concentration above 50 ppb. FeNO levels differs significantly between the farms but there was no difference between workers inside and outside greenhouses. Greenhouse workers at flower farms had higher prevalence of blocked nose than workers outside, which may indicate the presence of rhinitis. Endotoxin exposure was low. There were few workers with objective signs of airway inflammation; this might be because the mean working time in the

  19. 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. Copyright © 2013 American Dairy Science Association

  20. Occupational exposure to endotoxins and lung cancer risk: results of the ICARE Study.

    Science.gov (United States)

    Ben Khedher, Soumaya; Neri, Monica; Guida, Florence; Matrat, Mireille; Cenée, Sylvie; Sanchez, Marie; Menvielle, Gwenn; Molinié, Florence; Luce, Danièle; Stücker, Isabelle

    2017-09-01

    To investigate the role of occupational exposure to endotoxins in lung cancer in a French population-based case-control study (ICARE (Investigation of occupational and environmental causes of respiratory cancers)). Detailed information was collected on the occupational history and smoking habits from 2926 patients with histologically confirmed lung cancer and 3555 matched controls. We evaluated each subject's endotoxin exposure after cross referencing International Standard Classification of Occupations (ISCO) codes (for job tasks) and Nomenclature d'Activités Françaises (NAF) codes (for activity sectors). Endotoxin exposure levels were attributed to each work environment based on literature reports. ORs and 95% CIs were estimated using unconditional logistic regression models and controlled for main confounding factors. An inverse association between exposure to endotoxins and lung cancer was found (OR=0.80, 95% CI 0.66 to 0.95). Negative trends were shown with duration and cumulative exposure, and the risk was decreased decades after exposure cessation (all statistically significant). Lung cancer risk was particularly reduced among workers highly exposed (eg, in dairy, cattle, poultry, pig farms), but also in those weakly exposed (eg, in waste treatment). Statistically significant interactions were shown with smoking, and never/light smokers were more sensitive to an endotoxin effect than heavy smokers (eg, OR=0.14, 95% CI 0.06 to 0.32 and OR=0.80, 95% CI 0.45 to 1.40, respectively, for the quartiles with the highest cumulative exposure, compared with those never exposed). Pronounced inverse associations were shown with adenocarcinoma histological subtype (OR=0.37, 95% CI 0.25 to 0.55 in the highly exposed). Our findings suggest that exposure to endotoxins, even at a low level, reduces the risk of lung cancer. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is

  1. Risk of lung cancer in relation to contiguous windows of endotoxin exposure among female textile workers in Shanghai

    Science.gov (United States)

    Costello, Sadie; Applebaum, Katie M.; Ray, Roberta M.; Astrakianakis, George; Gao, Dao Li; Thomas, David B.; Checkoway, Harvey; Eisen, Ellen A.

    2012-01-01

    Objective Exposure to endotoxin has been consistently associated with a reduced risk of lung cancer. However, there is a paucity of information regarding temporal aspects of this relationship. The objective of this study was to investigate the associations between contiguous windows of endotoxin exposure and risk of lung cancer. Methods Data were reanalyzed from a case-cohort study (602 cases, 3,038 subcohort) of female textile workers in Shanghai, China. Cumulative endotoxin exposure was partitioned into two windows: ≥20 and windows. Results There was an inverse trend of decreasing risk of lung cancer associated with increasing levels of endotoxin exposure ≥20 years before risk (p trend = 0.02). Women in the highest two categories of cumulative exposures had hazard ratios of 0.78 (95% CI 0.60–1.03) and 0.77 (95% CI 0.58–1.02) for lung cancer, respectively, in comparison with unexposed textile workers. There was, however, a weaker association and not statistically significant between lung cancer and endotoxin exposure accumulated in the more recent window (<20 years before risk). Conclusion Results provide further evidence that endotoxin exposure that occurred 20 years or more before risk confers the strongest protection against lung cancer, indicating a possible early anti-carcinogenic effect. Further studies are needed to better understand the underlying biological mechanisms for this effect. PMID:21732048

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

    DEFF Research Database (Denmark)

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

    2002-01-01

    , endotoxin, 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......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 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...

  3. Evaluation of drought tolerance in different growth stages of maize ...

    African Journals Online (AJOL)

    Jane

    2011-10-12

    Oct 12, 2011 ... index; STI: stress tolerance index. condition. MO17 was a tolerant inbred line based on TOL and its low quantity indicates tolerant inbred lines (Table 4). MO17 was low yielding under all conditions. It seems that. TOL had succeeded in selecting genotypes with high yield under stress, but had failed to select ...

  4. 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.

  5. Limulus amebocyte lysate technique (LAL) for bacterial endotoxin control in radiodiagnosis agents (kits) and radioisotopes

    International Nuclear Information System (INIS)

    Morote, M.; Robles, A.; Ramos, B.; Otero, M.

    1997-01-01

    A procedure based on a fast technique of LAL individual kits has been devised to control bacterial endotoxins in radiodiagnosis agents (RDA): HEMTEC, DEIDA, PPI, AMD, GHCa, RENTEC, DMSA, MAA, TSC, HERTEC, DTPA, BRATEC and EDTMP as well as in radioisotopes I-131 and Tc99m. The procedures begins with the determination of the following values, injection volume (IV), endotoxin limits (EL), maximum valid dilution (MVD), total mass (TM), reconstitution volume (RV), concentration (mg/ml), and final dilution (FD). Subsequently, a procedure is carried out to conduct an 'in vitro' control of the radiodiagnosis agents and radioisotopes with LAL individual kits; the procedures includes: reconstitution of the sample to be controlled, dilution, inoculation of the diluted sample in LAL tubes and incubation at 37 o C for an hour. Finally, results are interpreted through the observation of gel formation or not in LAL tubes

  6. 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

    previously been shown to mediate cellular activation in both human and murine immune cells.Objective: We examined the response to different beta-lactoglobulin preparations in naive immune cells.Methods: Splenocytes and cells from mesenteric lymph nodes derived from BALB/c mice bred and maintained on a milk......-marrow-derived dendritic cells was examined.Results: We observed that some commercial beta-lactoglobulin preparations induced pronounced proliferation of both spleen cells and cells from mesenteric lymph nodes; production of TNF-alpha, IL-6, IL-1beta, and IL-10; and an increased level of intracellular glutathione...... 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...

  7. Use of limulus assay to compare the biological activity of peptidoglycan and endotoxin.

    Science.gov (United States)

    Wildfeuer, A; Heymer, B; Spilker, D; Schleifer, K H; Vanek, E; Haferkamp, O

    1975-07-01

    The gelation of a lysate prepared from the amebocytes of Limulus polyphemus, the horseshoe crab, has been used to detect endotoxin-like material in clinical studies and in "in vitro" experiments. The investigation of blood samples from 54 hospitalized patients suspected of having endotoxemia, revealed a positive limulus test in 14. Infections due to gram-positive organisms were not associated with positive assays. These results were in agreement with the observation that living gram-positive microorganisms - in contrast to gram-negative bacteria - did not initiate lysate gelation when studied "in vitro". Only very high concentrations of peptidoglycan, isolated from the cell walls of various gram-positive bacteria, induced the reaction. Therefore, these findings support the view, that the limulus test is relatively specific for the detection of gram-negative bacterial endotoxin and endotoxemia.

  8. Tachypleus lysate test for endotoxin in patients with Gram negative bacterial infections.

    Science.gov (United States)

    Usawattanakul, W; Tharavanij, S; Limsuwan, A

    1979-03-01

    Amoebocyte lysate from the horseshoe crabs (Tachypleus gigas) which abounds in the Gulf of Thailand was used to detect endotoxin in patients with Gram-negative bacteremia, in patients with Gram-positive bacterial infections as well as in the control. The Tachypleus lysate test (TLT) was positive in 94.4% of 36 patients with Gram-negative bacteremia before initiation of antibiotic therapy. Only 4% of 50 healthy individuals were positive and all 7 patients with Gram-positive bacterial infections were negative. The threshold sensitivity of TLT was 0.625 micrograms endotoxin per ml of the plasma. In comparison with the commercial Limulus lysate test (LLT), the TLT was slightly more sensitive in exhibiting higher grade of reaction, eventhough the threshold sensitivity was the same.

  9. Preparation of PVA/amino multi-walled carbon nanotubes nanocomposite microspheres for endotoxin adsorption.

    Science.gov (United States)

    Zong, Wenhui; Chen, Jian; Han, Wenyan; Cheng, Guanghui; Chen, Jie; Wang, Yue; Wang, Weichao; Ou, Lailiang; Yu, Yaoting; Shen, Jie

    2018-02-01

    A novel polyvinyl alcohol-amino multi-walled carbon nanotube (PVA-AMWCNT) nanocomposite microsphere was prepared successfully for the first time and used for endotoxin removal. The resulting AMWCNT modified PVA microsphere was characterized by SEM, Raman spectrum and fluorescence image, which indicated AMWCNT was dispersed into the macropores of PVA microsphere uniformly. The PVA-AMWCNT microspheres showed better adsorption capability and faster adsorption equilibrium for endotoxin in aqueous solution when compared to the PVA microsphere with polymyxin B (PMB) as ligand. More noteworthy, the PVA based microspheres had little nonspecific adsorption in simulated serum. Therefore, PVA-AMWCNT nanocomposite microsphere with an excellent haemocompatibility has a great potential application in clinical blood purification.

  10. The addition of submergence-tolerant Sub1 gene into high yielding MR219 rice variety and analysis of its BC2F3 population in terms of yield and yield contributing characters to select advance lines as a variety

    Directory of Open Access Journals (Sweden)

    Fahim Ahmed

    2016-09-01

    Full Text Available A cross was made between MR219 (high yielding but submergence intolerant and Swarna-Sub1 (submergence tolerant to produce submergence-tolerant rice variety using the marker-assisted backcrossing (MABC method to protect the farmers of low-lying land from flash floods during rain. Knowledge of yield and yield contributing factors plays a vital role in the selection process of a variety. This experim ent was designed to determine the genetic diversity among recently produced different lines of BC2F3 population and also to compare all the lines with MR219 to find the best one. Agronomical, yield and yield contributing data were taken, while genotypic and phenotypic coefficients, variance components and heritability were estimated. Introgression of the target gene, Sub1, was done using tightly linked marker, and also background recovery was measured using simple sequence repeat (SSR markers in different generations. The observed recurrent parent genome (RPG recovery of BC2F2 generation was 95.37%, which indicates high-level similarity between the recurrent parent (MR219 and the resulting lines. Thirty newly developed lines of BC2F3 population, resulting backcross of MR219 and Swarna-Sub1, were planted with four replications following randomised complete block design (RCBD. Newly developed lines were grouped into four clusters based on traits with UPGMA dendrogram and cluster analysis to select the 10 best plants. This study will help the future researchers to select the best plants of a breeding programme after introgression of a gene considering phenotype performances to develop new varieties.

  11. 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

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

    International Nuclear Information System (INIS)

    Guimaraes, B.G.; Garratt, R.C.; Oliva, G.; Lemos, M.V.F.; Arantes, O.M.N.

    1996-01-01

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

  13. Role of Bacterial Endotoxins of Intestinal Origin in Rat Heat Stress Mortality

    Science.gov (United States)

    1982-01-19

    the effect of increased survival associated Received 19 January 1982; accepted in final form 15 July 1982. REFERENCES 1. ALTURA , B. M., AND S. G...Limulus assay for endotoxin. N. Engi. J. Med. 289: 484- 1414-1419, 1968. 485, 1973. 2. ALTURA , B. M., AND S. G. HERSHEY. Acute intestinal ischemia 9...and chronic lung injury. J. App. Physiol: Soc. 10: 361-371, 1971. Respirat. Environ. Exercise Physiol. 47: 577-581, 1979. 3. ALTURA , B. M., C. THAW

  14. Effect of vasopressors on organ blood flow during endotoxin shock in pigs

    International Nuclear Information System (INIS)

    Breslow, M.J.; Miller, C.F.; Parker, S.D.; Walman, A.T.; Traystman, R.J.

    1987-01-01

    A volume-resuscitated porcine endotoxin shock model was used to evaluate the effect on organ blood flow of increasing systemic arterial blood pressure with vasopressors. Administration of 0.05-0.2 mg/kg of Escherichia coli endotoxin (E) reduced mean arterial blood pressure (MAP) to 50 mmHg, decreased systemic vascular resistance to 50% of control, and did not change cardiac output or heart rate. Blood flow measured with radiolabeled microspheres to brain, kidney, spleen, and skeletal muscle was reduced during endotoxin shock, but blood flow to left ventricle, small and large intestine, and stomach remained at pre-endotoxin levels throughout the study period. Four groups of animals were used to evaluate the effect of vasopressor therapy. Vasopressors were administered starting 60 min after E exposure, and the dose of each was titrated to increase MAP to 75 mmHg. Despite the increase in MAP, brain blood flow did not increase in any group. Norepinephrine alone increased blood flow to the left ventricle. The dose of norepinephrine required to increase MAP by 20-25 mmHg during E shock was 30 times the does required for a similar increase in MAP in animals not receiving E. The authors conclude 1) that hypotension in the fluid resuscitated porcine E shock model is primarily the result of peripheral vasodilatation, 2) that the vascular response to vasoconstrictors in this model is markedly attenuated following E administration, 3) that blood pressure elevation with norepinephrine, dopamine, and phenylephrine neither decreases blood flow to any organs nor increases blood flow to organs with reduced flow, and 4) that norepinephrine, dopamine, and phenylephrine affect regional blood flow similarly in this model

  15. Alterations in hepatic gluconeogenic amino acid uptake and gluconeogenesis in the endotoxin treated conscious dog.

    Science.gov (United States)

    Meinz, H; Lacy, D B; Ejiofor, J; McGuinness, O P

    1998-04-01

    We examined the effect of a 240 min intraportal infusion of a nonlethal dose of Escherichia coli endotoxin (.21 g x kg(-1) x min[-1]) on hepatic amino acid and glucose metabolism in chronically catheterized 42 h fasted conscious dogs (n = 8). Hepatic metabolism was assessed using tracer (3-[3H]glucose [U-14C]alanine) and arteriovenous difference techniques. After endotoxin administration net hepatic glucose output increased twofold. Arterial plasma insulin levels decreased by 25%, whereas arterial plasma glucagon and cortisol levels increased 10- and 6-fold, respectively. Arterial lactate levels increased 6.4-fold, whereas net hepatic lactate uptake was not increased. Arterial alanine levels (1.6-fold) and net hepatic alanine uptake (1.3-fold) increased, whereas net hepatic alanine fractional extraction was unaltered. In contrast, the arterial levels of the other gluconeogenic amino acids (glutamine, glycine, serine, and threonine) decreased. Despite this decrease, net uptake of these amino acids by the liver did not decrease, because net hepatic amino acid fractional extraction increased. Total net hepatic gluconeogenic precursor uptake was unaltered (1.1 +/- .1 to 1.3 +/- .3 mg x kg(-1) x min(-1) expressed in glucose equivalents). In summary, gluconeogenesis does not increase after endotoxin administration. Thus, an increase in net hepatic glycogenolysis accounts for the majority of the increase in hepatic glucose production. The lack of an increase in alanine fractional extraction, despite hyperglucagonemia and a rise in the fractional extraction of other gluconeogenic amino acids, suggests that endotoxin specifically impairs hepatic alanine entry in vivo.

  16. Application of Recombinant Factor C Reagent for the Detection of Bacterial Endotoxins in Pharmaceutical Products.

    Science.gov (United States)

    Bolden, Jay; Smith, Kelly

    2017-01-01

    Recombinant Factor C (rFC) is non-animal-derived reagent used to detect bacterial endotoxins in pharmaceutical products. Despite the fact that the reagent was first commercially available nearly 15 years ago, the broad use of rFC in pharmaceutical industry has long been lagging, presumably due to historical single-source supplier concerns and the lack of inclusion in worldwide pharmacopeias. Commercial rFC reagents are now available from multiple manufacturers, thus single sourcing is no longer an issue. We report here the successful validation of several pharmaceutical products by an end-point florescence-based endotoxin method using the rFC reagent. The method is equivalent or superior to the compendia bacterial endotoxins test method. Based on the comparability data and extenuating circumstances, the incorporation of the end point fluorescence technique and rFC reagent in global compendia bacterial endotoxins test chapters is desired and warranted. LAY ABSTRACT: Public health has been protected for over 30 years with the use of a purified blood product of the horseshoe crab, limulus amebocyte lysate. More recently, this blood product can be produced in biotech manufacturing processes, which reduces potential impacts to the horseshoe crab and related species dependent upon the crab, for example, migrating shorebirds. The pharmaceutical industry has been slow to adopt the use of this reagent, Recombinant Factor C (rFC), for various reasons. We evaluated the use of rFC across many pharmaceutical products, and in other feasibility demonstration experiments, and found rFC to be a suitable alternative to the animal-derived limulus amebocyte lysate. Incorporation of rFC and its analytical method into national testing standards would provide an equivalent or better test while continuing to maintain patient safety for those who depend on medicines and while securing pharmaceutical supply chains. In addition, widespread use of this method would benefit existing animal

  17. Fourier Transform Infrared Spectroscopy as a Tool in Analysis of Proteus mirabilis Endotoxins.

    Science.gov (United States)

    Żarnowiec, Paulina; Czerwonka, Grzegorz; Kaca, Wiesław

    2017-01-01

    Fourier transform infrared spectroscopy (FT-IR) was used to scan whole bacterial cells as well as lipopolysaccharides (LPSs, endotoxins) isolated from them. Proteus mirabilis cells, with chemically defined LPSs, served as a model for the ATR FT-IR method. The paper focuses on three steps of infrared spectroscopy: (1) sample preparation, (2) IR scanning, and (3) multivariate analysis of IR data (principal component analysis, PCA).

  18. 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....

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

    OpenAIRE

    Yung-Ray Hsu; Shu-Wen Chang; Chang-Hao Yang; Yi-An Lee; Tzu-Yun Kao

    2016-01-01

    Purpose. The transcellular arginine transportation via cationic amino acid transporter (CAT) is the rate-limiting step in nitric oxide (NO) synthesis, which is crucial in intraocular inflammation. In this study, CAT isoforms and inducible nitric oxide synthase (iNOS) expression was investigated in endotoxin-induced uveitis (EIU). Methods. EIU was induced in Lewis rats by lipopolysaccharide (LPS) injection. In the treatment group, the rats were injected intraperitoneally with the proteasome in...

  20. Elevated Endotoxin Levels in Human Intravenous Immunoglobulin Concentrates Caused by (1->3)-{beta}-D-Glucans.

    Science.gov (United States)

    Buchacher, Andrea; Krause, Dagmar; Wiry, Gerda; Weinberger, Josef

    2010-01-01

    Endotoxins have been measured routinely in the final product and during the production process to produce non-pyrogenic parenterals. Limulus-amoebocyte-lysate-reactive material was found in in-process samples and final product of one of Octapharma's intravenous immunoglobulin (IVIG) preparations. Limulus-amoebocyte-lysate (LAL) is activated by bacterial endotoxins and by (1→3)-β-D-glucans. The contribution of both compounds on the LAL-related signal was determined by three different approaches: (1) using a test specific for (1→3)-β-D-glucans, (2) by addition of β-glucan blocker, and (3) by the use of a recombinant endotoxin assay. It was shown that none of our IVIG concentrates contained elevated endotoxin contents but that the higher LAL reaction could be ascribed to (1→3)-β-D-glucans extracted from cellulose filter pads. The use of an endotoxin test kit highly sensitive for (1→3)-β-D-glucans might lead to false-positive results. (1→3)-β-D-glucans spike solutions did not evoke an increase of temperature in rabbits, suggesting that a pyrogenic reaction is not expected in patients. Endotoxins have been measured routinely in the final product and during the production process to produce non-pyrogenic parenterals. Limulus-amoebocyte-lysate-reactive material was found in in-process samples and final product of one of Octapharma's intravenous immunoglobulin (IVIG) preparations. Limulus-amoebocyte-lysate (LAL) is activated by bacterial endotoxins and by (1→3)-β-D-glucans. The contribution of both compounds on the LAL-related signal was determined by three different approaches: (1) using a test specific for (1→3)-β-D-glucans, (2) by addition of β-glucan blocker, and (3) by the use of a recombinant endotoxin assay. It has been shown that none of our IVIG concentrates contained elevated endotoxin contents but that the higher LAL reaction could be ascribed to (1→3)-β-D-glucans extracted from cellulose filter pads. The use of an endotoxin test kit

  1. Scanning ion-selective electrode technique and X-ray microanalysis provide direct evidence of contrasting Na+ transport ability from root to shoot in salt-sensitive cucumber and salt-tolerant pumpkin under NaCl stress.

    Science.gov (United States)

    Lei, Bo; Huang, Yuan; Sun, Jingyu; Xie, Junjun; Niu, Mengliang; Liu, Zhixiong; Fan, Molin; Bie, Zhilong

    2014-12-01

    Grafting onto salt-tolerant pumpkin rootstock can increase cucumber salt tolerance. Previous studies have suggested that this can be attributed to pumpkin roots with higher capacity to limit the transport of Na(+) to the shoot than cucumber roots. However, the mechanism remains unclear. This study investigated the transport of Na(+) in salt-tolerant pumpkin and salt-sensitive cucumber plants under high (200 mM) or moderate (90 mM) NaCl stress. Scanning ion-selective electrode technique showed that pumpkin roots exhibited a higher capacity to extrude Na(+), and a correspondingly increased H(+) influx under 200 or 90 mM NaCl stress. The 200 mM NaCl induced Na(+)/H(+) exchange in the root was inhibited by amiloride (a Na(+)/H(+) antiporter inhibitor) or vanadate [a plasma membrane (PM) H(+) -ATPase inhibitor], indicating that Na(+) exclusion in salt stressed pumpkin and cucumber roots was the result of an active Na(+)/H(+) antiporter across the PM, and the Na(+)/H(+) antiporter system in salt stressed pumpkin roots was sufficient to exclude Na(+) X-ray microanalysis showed higher Na(+) in the cortex, but lower Na(+) in the stele of pumpkin roots than that in cucumber roots under 90 mM NaCl stress, suggesting that the highly vacuolated root cortical cells of pumpkin roots could sequester more Na(+), limit the radial transport of Na(+) to the stele and thus restrict the transport of Na(+) to the shoot. These results provide direct evidence for pumpkin roots with higher capacity to limit the transport of Na(+) to the shoot than cucumber roots. © 2014 Scandinavian Plant Physiology Society.

  2. 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.

  3. 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

  4. Endotoxin levels correlate positively with a sedentary lifestyle and negatively with highly trained subjects

    Directory of Open Access Journals (Sweden)

    Oyama Lila M

    2010-08-01

    Full Text Available Abstract Introduction A sedentary lifestyle increases the risk of developing cardiovascular disease, obesity, and diabetes. This phenomenon is supported by recent studies suggesting a chronic, low-grade inflammation status. Endotoxin derived from gut flora may be key to the development of inflammation by stimulating the secretion of inflammatory factors. This study aimed to examine plasma inflammatory markers and endotoxin levels in individuals with a sedentary lifestyle and/or in highly trained subjects at rest. Methods: Fourteen male subjects (sedentary lifestyle n = 7; highly trained subjects n = 7 were recruited. Blood samples were collected after an overnight fast (~12 h. The plasmatic endotoxin, plasminogen activator inhibitor type-1 (PAI-1, monocyte chemotactic protein-1 (MCP1, ICAM/CD54, VCAM/CD106 and lipid profile levels were determined. Results Endotoxinemia was lower in the highly trained subject group relative to the sedentary subjects (p Conclusion These results indicate that a lifestyle associated with high-intensity and high-volume exercise induces favorable changes in chronic low-grade inflammation markers and may reduce the risk for diseases such as obesity, diabetes and cardiovascular diseases.

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

    Science.gov (United States)

    Mizumura, Hikaru; Ogura, Norihiko; Aketagawa, Jun; Aizawa, Maki; Kobayashi, Yuki; Kawabata, Shun-Ichiro; Oda, Toshio

    2017-02-01

    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.

  6. Effect of inhaled endotoxin on mucociliary clearance and airway inflammation in mild smokers and nonsmokers.

    Science.gov (United States)

    Bennett, William D; Alexis, Neil E; Almond, Martha; Herbst, Margaret; Zeman, Kirby L; Peden, David B

    2014-12-01

    In healthy nonsmokers, inhaled endotoxin [lipopolysaccharide (LPS)] challenge induces airway neutrophilia and modifies innate immune responses, but the effect on mucociliary clearance (MCC), a key host defense response, is unknown. Although smokers are chronically exposed to LPS through inhaled tobacco smoke, the acute effect of inhaled LPS on both MCC and airway inflammation is also unknown. The purpose of this study was to determine the effect of inhaled LPS on MCC in nonsmokers and mild smokers with normal pulmonary function. We performed an open-label inhalational challenge with 20,000 endotoxin units in healthy adult nonsmokers (n=18) and young adult, mild smokers (n=12). At 4 hr post LPS challenge, we measured MCC over a period of 2 hr, followed by sputum induction to assess markers of airway inflammation. No significant changes in spirometry occurred in either group following LPS challenge. Following LPS, MCC was significantly (psmokers [MCC=10±9% (challenge) vs. 15±8% (baseline), MCC=14±9% (challenge) vs. 16±10% (baseline), respectively]. Both groups showed a significant (psmokers is unaffected by mild endotoxin challenge, likely due to preexisting effects of cigarette smoke on their airway epithelium.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2016-03-01

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

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

    International Nuclear Information System (INIS)

    Norbäck, Dan; Hashim, Jamal Hisham; Markowicz, Pawel; Cai, Gui-Hong; Hashim, Zailina; Ali, Faridah; Larsson, Lennart

    2016-01-01

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

  9. A 15-week experimental exposure of pigs to airborne dust with added endotoxin in a continuous flow exposure chamber.

    Science.gov (United States)

    Jolie, R; Bäckström, L; Olson, L; Chase, C

    1999-04-01

    The purpose of this study was to evaluate the effect of longterm exposure to airborne dust and endotoxin on the respiratory system of pigs. A continuous flow exposure chamber was built for the purpose of exposing pigs to selected airborne contaminants. Pigs (n = 6) were exposed to a combination of a very fine corn/soybean meal (40.6 mg/m3) with added lipopolysaccharide (LPS; 12.4 microg/m3) for 8 h/d over 5 d for 15 wk (75 d of exposure). Control pigs (n = 6) were housed in a room with minimal contamination of these airborne contaminants. Surprisingly, dust in the exposure chamber and the control room was highly contaminated with peptidoglycan. Changes in the lung were monitored by collecting bronchoalveolar lavage (BAL) fluid for cytology at 5 different time points throughout the exposure period. Blood samples were collected at the same time for hematology. A non-specific respiratory inflammatory response was found in exposed and control pigs, as suggested by the increased neutrophils in BAL fluid and the small inflammatory areas in the lung tissue. No macroscopic lung lesions were observed in control or exposed pigs. The findings in the control pigs imply that even low dust concentrations and possibly peptidoglycan contamination can induce cellular changes in the BAL fluid and that a true control pig does not exist. In addition, the exposed pigs developed a mild eosinophilia, indicating an allergic response to the airborne contaminants.

  10. Endotoxin-induced and vaccine-induced systemic inflammation both impair endothelium-dependent vasodilation, but not pulse wave reflection

    Directory of Open Access Journals (Sweden)

    Lind L

    2012-07-01

    Full Text Available Lars Lind,1 Johannes Hulthe,2,3 Annika Johansson,3 Ewa Hedner31Department of Medicine, University Hospital, Uppsala, 2Sahlgrenska Hospital, Gothenburg, 3AstraZeneca Research and Development, Mölndal, SwedenBackground: Inflammation induced by either endotoxin or vaccination has previously been shown to impair endothelium-dependent vasodilation (EDV in healthy young individuals. However, the vascular effects of these two mechanisms of inducing inflammation have not been compared in the same individuals.Methods: Twelve young healthy males were studied at the same time of the day on three occasions in a random order; on one occasion 4 hours following an endotoxin injection (Escherichia coli endotoxin, 20 IU/kg, on another occasion 8 hours following vaccination against Salmonella typhi, and on a third occasion 4 hours following a saline control injection. EDV and endothelium-independent vasodilation (EIDV were evaluated by local infusions of acetylcholine and sodium nitroprusside in the brachial artery, and forearm blood flow was measured with venous occlusion plethysmography. The augmentation index was determined by pulse wave analysis as an index of pulse wave reflection.Results: Both endotoxin and vaccination impaired EDV to a similar degree compared with the saline control (P = 0.005 and P = 0.014, respectively. EIDV was not significantly affected by inflammation. Endotoxin, but not vaccination, increased body temperature and circulating levels of intracellular adhesion molecule-1 and interleukin-6. Augmentation index was not affected by the interventions.Conclusion: Despite the fact that endotoxin induced a more pronounced degree of inflammation than vaccination, both inflammatory challenges impaired EDV to a similar degree, supporting the view that different inflammatory stimuli could induce harmful effects on the vasculature.Keywords: endothelium, endotoxin, vaccination, vasodilation, inflammation

  11. The specificity of Sushi peptides for endotoxin and anionic phospholipids: potential application of POPG as an adjuvant for anti-LPS strategies.

    Science.gov (United States)

    Li, P; Sun, M; Ho, B; Ding, J L

    2006-04-01

    Sushi peptides [S1 (Sushi 1 peptide) and S3] are derived from the LPS (lipopolysaccharide; also known as endotoxin)-binding domains of an LPS-sensitive serine protease, Factor C, from the horseshoe crab (Carcinoscorpius rotundicauda). S1 and S3 interact at high affinity with LPS. The intermolecular disulphide bonding in the S3 dimer is indispensable for its LPS binding, disruption and consequent neutralization. Simultaneously, the specific interaction between the Sushi peptides and bacterial membrane phospholipids further explains the selective propensity of these peptides for the gram-negative bacteria. Our findings yield insights into a complex molecular paradigm in which the juxtaposition of LPS molecules and the anionic phospholipid POPG (1-palmitoyl-2-oleoyl phosphatidylglycerol) on the bacterial outer membrane confers such interfacial properties which create the optimal environment for the interaction between the peptides and bacterial membrane lipids.

  12. Mutational tolerance to carbendazim in Botrytis cinerea

    International Nuclear Information System (INIS)

    Geeson, J.D.

    1978-01-01

    No spontaneous mutation for tolerance to the fungicide carbendazim was detected in c. 10 8 conidia from each of eight carbendazim-sensitive field isolates of Botrytis cinerea. Conidia of B. cinerea were highly insensitive to u.v.-irradiation, although after severe irradiation treatments mutant strains showing the same levels of tolerance as two groups of carbendazim-tolerant field isolates were selected at frequencies of between 10 -9 and 10 -6 of survivors. Mutants with low levels of tolerance (EDsub(50)( -1 carbendazim, 'partially-tolerant') were selected from irradiated conidia obtained from sensitive field isolates and a further series of mutants capable of growth on 10,000 μg ml -1 carbendazim ('fully-tolerant') were selected from irradiated conidia from either partially-tolerant mutants or from partially tolerant field isolates. Both mutation steps were confirmed in similar experiments in which tolerance to an unrelated fungicide, 2,6-dichloro-4-nitroaniline (DCNA), was incorporated as a genetic marker in the parent strains. (author)

  13. Immunization of cows with novel core glycolipid vaccine induces anti-endotoxin antibodies in bovine colostrum.

    Science.gov (United States)

    Cross, Alan S; Karreman, Hubert J; Zhang, Lei; Rosenberg, Zeil; Opal, Steven M; Lees, Andrew

    2014-10-21

    Translocation of gut-derived Gram-negative bacterial (GNB) lipopolysaccharide (LPS, or endotoxin) is a source of systemic inflammation that exacerbates HIV, cardiovascular and gastrointestinal diseases and malnutrition. The oral administration of bovine colostrum (BC) reduces endotoxemia in patients with impaired gut barrier function. Consequently, BC enriched in antibodies to LPS may ameliorate endotoxemia-related morbidities. We developed a detoxified J5 LPS/group B meningococcal outer membrane protein (J5dLPS/OMP) vaccine that induces antibodies against a highly conserved core region of LPS and protects against heterologous GNB infection. We now examine the ability of this vaccine to elicit anti-core endotoxin antibodies in BC. Two cohorts of pregnant cows were immunized with this vaccine in combination with FICA (Cohort 1) or Emulsigen-D (Cohort 2) adjuvants. Antibody responses to the J5 core LPS antigen were measured in both serum and colostrum and compared to antibody levels elicited by a commercially available veterinary vaccine (J5 Bacterin) comprised of heat-killed Escherichia coli O111, J5 mutant bacteria, from which the J5 LPS was purified. The J5dLPS/OMP vaccine induced high titers of serum IgG antibody to J5 LPS in all seven cows. Both IgG and to a lesser extent IgA anti-J5 LPS antibodies were generated in the colostrum. The J5dLPS/OMP vaccine was significantly more immunogenic in mice than was the J5 Bacterin. BC enriched in anti-J5 LPS antibody reduced circulating endotoxin levels in neutropenic rats, a model of "leaky gut". The J5dLPS/OMP vaccine elicits high titers of serum anti-endotoxin antibodies in cows that is passed to the colostrum. This BC enriched in anti-core LPS antibodies has the potential to reduce endotoxemia and ameliorate endotoxin-related systemic inflammation in patients with impaired gut barrier function. Since this vaccine is significantly more immunogenic than the J5 Bacterin vaccine, this J5dLPS/OMP vaccine might prove to be

  14. Sensitive measurement of endotoxin by radio-rocket immunoelectrophoresis using [125I]Staphylococcus aureus protein A

    International Nuclear Information System (INIS)

    Stevens, P.; Alam, S.; Young, L.S.; Chesebro, K.

    1981-01-01

    Antibody directed against the core glycolipid antigen (CGL) of the mutant Salmonella minnesota Re 595 has been shown to cross-react with endotoxin from bacteria within the group Enterobacteriaceae. Using this cross-reactive CGL antibody the authors have developed a sensitive (250 pg) radio-rocket immunoelectrophoretic technique to measure endotoxin. They used the principles of rocket immunoelectrophoresis and increased the sensitivity by using 125 I-labelled staphylococcal protein A which serves as a sensitive probe to bind to the Fc portion of the IgG complexed with antigen. The rocket-shaped [ 125 I]protein A labelled immune complexes were detected by radioautography. The sensitivity is 100-fold greater than conventional Coomassie brilliant blue staining. Measurement of CGL was inhibited by normal human serum. However, the assay had the capacity to quantitate endotoxin in buffer extracts of clinically isolated Escherichia coli, Serratia marcescens, Klebsiella pneumoniae but not Pseudomonas aeruginosa. Analysis of various preparations of CGL obtained from different investigators demonstrated wide variation in their immunoreactivity. Because of the significant cross-reaction to detect various endotoxins this method has the potential to measure endotoxemia and assess the immunochemical quality of various endotoxin preparations. Additionally, the techniques of using [ 125 I]protein A has wide applicability for the sensitive measurement of other antigens. (Auth.)

  15. Measurement of endotoxin levels in blood of hemodialysis Patients by 'Lal' test and comparision of its efficacy with blood culture

    Directory of Open Access Journals (Sweden)

    Gh Vazirzadeh

    2006-01-01

    Full Text Available Introduction: Presently, bacteremia is the principal cause of morbidity in patients undergoing hemodialysis. Gram-negative bacteria account for approximately 50 percent of documented infections. Endotoxins released during lysis of gram negative bacteremia result in inflammatory and defense response by the body and if not treated promptly result in septic shock and ultimately death of the patient. This study describes the detection of endotoxins in blood of patients with bacteremia due to gram - negative bacteria by LAL test. Method: Blood samples of 278 hemodialysis patients were analyzed in this study and pathogens were isolated from blood culture samples. Then, their antibiotic sensitivity was determined. In patients with positive blood culture, endotoxin levels were measured by LAL-test. Results: Frequency of bacteremia in patients was 13.6% . The prevalence of gram – negative bacteremia was 44.7%. E coli were the major pathogens, while staphylococcus aureus was the most common gram positive bacterium. Endotoxin was detected in 15 patients (3.8 ± 1.08 EU/ml . The sensitivity and specificity of endotoxins for gram – negative bacteremia were 88% and 95%, respectively. Conclusion: The results indicate that the LAL method is a fast, sensitive and simple method. There was no significant difference between the results of blood culture and LAL – test ( P > 0.05 .

  16. COMPARISON OF EFFICACY AND TOLERABILITY OF TWO SELECTIVE M3 RECEPTOR ANTAGONISTS SOLIFENACIN AND DARIFENACIN IN WOMEN WITH OVERACTIVE BLADDER – THE SOLIDAR STUDY

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    Igor But

    2018-02-01

    Full Text Available Background. To assess clinical efficacy and tolerance of solifenacin and darifenacin in women with OAB with urgency as primary end point. To estimate the impact of anticholinergic drugs on quality of life of patients and to assess treatment outcome (objective and subjective improvement. Methods. Multicentric, prospective, randomized, head to head, open label pilot study in 100 women with OAB. Patients were randomly assigned into two groups, 50 of them received solifenacine and the remaining 50 patients darifenacin. Study duration was 3 months, patients came to the office 1 and 3 months after inclusion in the study and they always filled out voiding diaries, urgency perception questionnaires as well as the Urogenital Distress Inventory (UDI and Incontinence Impact Questionnaires (IIQ. At the end of the study all patients estimated the success of treatment on the basis of VAS scale. Results. Only 77 patients were enrolled into study. Their average age was 54.8 years, and their BMI amounted to 27.6 kg/m2. The average duration of OAB symptoms and urge urinary incontinence was 86.0 and 46.5 months, respectively. 40 patients received solifenacin and remaining 37 patients darifenacin. Before treatment patients from both groups did not differ statistically significant in any of observed variables. 16 patients did not finish the study (8 solifenacin, 8 darifenacin, mainly due to side effects (8 patients. After 3 months of treatment we observed a significant improvement in severity, frequency and botherness of urgency in all patients, significantly improved were also other OAB symptoms (Table 1. Objective improvement after 1 months was greater in patients with solifenacin (p = 0.033, these patients also experienced significantly less irritative symptoms (p = 0.034. There was no difference in subjective improvement score between both drugs. Quality of life was better after three months in both groups of patients and there was no significant difference

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

    Science.gov (United States)

    Wan, Gwo-Hwa; Yan, Dah-Chin; Tung, Tao-Hsin; Tang, Chin-Sheng; Liu, Chiu-Hsin

    2013-01-01

    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.

  18. Evidence of divergent selection for drought and cold tolerance at landscape and local scales in Abies alba Mill. in the French Mediterranean Alps.

    Science.gov (United States)

    Roschanski, Anna M; Csilléry, Katalin; Liepelt, Sascha; Oddou-Muratorio, Sylvie; Ziegenhagen, Birgit; Huard, Frédéric; Ullrich, Kristian K; Postolache, Dragos; Vendramin, Giovanni G; Fady, Bruno

    2016-02-01

    Understanding local adaptation in forest trees is currently a key research and societal priority. Geographically and ecologically marginal populations provide ideal case studies, because environmental stress along with reduced gene flow can facilitate the establishment of locally adapted populations. We sampled European silver fir (Abies alba Mill.) trees in the French Mediterranean Alps, along the margin of its distribution range, from pairs of high- and low-elevation plots on four different mountains situated along a 170-km east-west transect. The analysis of 267 SNP loci from 175 candidate genes suggested a neutral pattern of east-west isolation by distance among mountain sites. F(ST) outlier tests revealed 16 SNPs that showed patterns of divergent selection. Plot climate was characterized using both in situ measurements and gridded data that revealed marked differences between and within mountains with different trends depending on the season. Association between allelic frequencies and bioclimatic variables revealed eight genes that contained candidate SNPs, of which two were also detected using F(ST) outlier methods. All SNPs were associated with winter drought, and one of them showed strong evidence of selection with respect to elevation. Q(ST)-F(ST) tests for fitness-related traits measured in a common garden suggested adaptive divergence for the date of bud flush and for growth rate. Overall, our results suggest a complex adaptive picture for A. alba in the southern French Alps where, during the east-to-west Holocene recolonization, locally advantageous genetic variants established at both the landscape and local scales. © 2015 John Wiley & Sons Ltd.

  19. Effect of Endotoxin from Serratia marcescens on the Permeability of Vessels in Hepatomas and Carrageenin Granulomas of Rats

    Science.gov (United States)

    Oswald, N. T. A.; Cater, D. B.

    1969-01-01

    Serratia marcescens polysaccharide (endotoxin) was injected i.v. into rats bearing carrageenin-induced granulomas and transplanted hepatomas, and the vascular-permeability changes were studied by injecting Pelikan ink i.v. at various times after the endotoxin (or saline in the controls). When the ink was given at 0 or 1 hr. there was little difference between the treated rats and the controls. But when the ink was given 2 or more hr. after the endotoxin there was a marked increase of the carbon-lined capillaries and venules, compared with controls, in tumour, granuloma, glomeruli, adrenal cortex, lung, lymph-nodes and sometimes heart. The significance of these changes is discussed in relation to the Shwartzman phenomenon, Arthus reaction, and adrenaline-induced vascular spasm producing anoxia. ImagesFigs. 2-5Figs. 6-9 PMID:4304259

  20. Detection of endotoxin in the plasma of patients with gram-negative bacterial sepsis by the Limulus amoebocyte lysate assay.

    Science.gov (United States)

    Pearson, F C; Dubczak, J; Weary, M; Bruszer, G; Donohue, G

    1985-01-01

    A total of 120 Limulus amoebocyte lysate (LAL) determinations were made on plasma obtained from normal, healthy human blood donors. Results demonstrated a mean endotoxin level in blood of 0.02 to 1.57 pg/ml. The amount of Escherichia coli endotoxin added to human plasma samples can be quantitated by both nephelometry and turbidimetry. Endotoxin-spiked samples were shown to be significantly different from unspiked samples. When plasma samples were collected from 45 patients hospitalized at three centers, a strong association was demonstrated between a positive Limulus amoebocyte lysate assay and a septic condition. Sensitivity, specificity, and false-positive and false-negative rates for the Limulus amoebocyte lysate assay as a diagnostic test for gram-negative bacteremia were estimated. PMID:4008617

  1. Host defense peptides of thrombin modulate inflammation and coagulation in endotoxin-mediated shock and Pseudomonas aeruginosa sepsis

    DEFF Research Database (Denmark)

    Kalle, Martina; Papareddy, Praveen; Kasetty, Gopinath

    2012-01-01

    Gram-negative sepsis is accompanied by a disproportionate innate immune response and excessive coagulation mainly induced by endotoxins released from bacteria. Due to rising antibiotic resistance and current lack of other effective treatments there is an urgent need for new therapies. We here...... present a new treatment concept for sepsis and endotoxin-mediated shock, based on host defense peptides from the C-terminal part of human thrombin, found to have a broad and inhibitory effect on multiple sepsis pathologies. Thus, the peptides abrogate pro-inflammatory cytokine responses to endotoxin...... sepsis, the peptide GKY25, while mediating a modest antimicrobial effect, significantly inhibited the pro-inflammatory response, decreased fibrin deposition and leakage in the lungs, as well as reduced mortality. Taken together, the capacity of such thrombin-derived peptides to simultaneously modulate...

  2. Treatment of rat Walker-256 carcinosarcoma with photodynamic therapy and endotoxin irradiated with high-energy electrons

    Science.gov (United States)

    Dima, Vasile F.; Vasiliu, Virgil V.; Laky, Dezideriu; Ionescu, Mircea D.; Dima, Stefan V.

    1994-03-01

    Experiments were performed on five batches of Wistar inbred rats with Walker-256 carcinosarcoma receiving sole treatment photodynamic therapy (PDT), irradiated endotoxin (R-LPS), native-endotoxin (N-LPS), or associated therapy (PDT + R-LPS) and the control batch (saline) consisted of animals with untreated Walker-256 tumors. The results were as follows: the sole treatment (PDT, R-LPS, N-LPS) gave survival rates between 56.3 and 60.7% and cure rates ranging from 32.1 to 37.5%. The `combined' therapy in multiple doses increased significantly (88.6%) the survival rate of tumor bearing rats as well as the highest incidence of complete tumor regression (71.4%). This work demonstrates that `combined' photodynamic and immunotherapy with irradiated endotoxin stimulates cell-mediated antitumoral activity and induces changes in the tumoral incidence in Walker-256 carcinosarcoma in the rat model.

  3. Concentration of airborne Staphylococcus aureus (MRSA and MSSA), total bacteria, and endotoxins in pig farms.

    Science.gov (United States)

    Masclaux, Frederic G; Sakwinska, Olga; Charrière, Nicole; Semaani, Eulalia; Oppliger, Anne

    2013-06-01

    Pigs are very often colonized by Staphylococcus aureus and transmission of such pig-associated S. aureus to humans can cause serious medical, hygiene, and economic problems. The transmission route of zoonotic pathogens colonizing farm animals to humans is not well established and bioaerosols could play an important role. The aim of this study was to assess the potential occupational risk of working with S. aureus-colonized pigs in Switzerland. We estimated the airborne contamination by S. aureus in 37 pig farms (20 nursery and 17 fattening units; 25 in summer, 12 in winter). Quantification of total airborne bacterial DNA, airborne Staphylococcus sp. DNA, fungi, and airborne endotoxins was also performed. In this experiment, the presence of cultivable airborne methicillin-resistant S. aureus (MRSA) CC398 in a pig farm in Switzerland was reported for the first time. Airborne methicillin-sensitive S. aureus (MSSA) was found in ~30% of farms. The average airborne concentration of DNA copy number of total bacteria and Staphylococcus sp. measured by quantitative polymerase chain reaction was very high, respectively reaching values of 75 (± 28) × 10(7) and 35 (± 9.8) × 10(5) copy numbers m(-3) in summer and 96 (± 19) × 10(8) and 40 (± 12) × 10(6) copy numbers m(-3) in winter. Total mean airborne concentrations of endotoxins (1298 units of endotoxin m(-3)) and fungi (5707 colony-forming units m(-3)) exceeded the Swiss recommended values and were higher in winter than in summer. In conclusion, Swiss pig farmers will have to tackle a new emerging occupational risk, which could also have a strong impact on public health. The need to inform pig farmers about biological occupational risks is therefore crucial.

  4. The cardiocirculatory and metabolic effects of endotoxin challenge after canine resuscitated hemorrhagic shock.

    Science.gov (United States)

    Horton, J

    1989-02-01

    Although adequate volume resuscitation has decreased mortality from hemorrhagic shock, recovery in many patients is complicated by sepsis. To determine whether a subject debilitated by hemorrhagic shock would exhibit greater cardiocirculatory dysfunction when challenged with sepsis, ten dogs (Group I) were hemorrhaged to a mean arterial blood pressure of 30 mm Hg. After 2 hours of hypotension, shed blood and lactated Ringer's solution (50 ml/kg) were given, and the dogs were observed for 3 to 6 days. Ten dogs were sham hemorrhage and served as controls (Group II). On the experimental day, all cardiovascular and hemodynamic parameters were measured in both groups of animals before endotoxin challenge. There was no significant difference in cardiac output, stroke volume, stroke work, +dP/dt max, myocardial blood flow, myocardial oxygen metabolism, or acid-base balance in the two groups. Compared to sham-hemorrhaged dogs, resuscitated shock dogs had a significantly lower mean arterial blood pressure (127 +/- 7 vs. 110 +/- 6 mm Hg; p less than 0.05), and heart rate was significantly higher (86 +/- 6 vs. 109 +/- 7 beats/minute; p less than 0.05). Furthermore, maximal rate of left ventricular pressure fall (-dP/dT max) was significantly lower in the animals previously hemorrhaged, suggesting a persistent defect in left ventricular relaxation. Blood glucose and insulin levels were significantly elevated in the resuscitated shocked dogs, likely due to increased circulating catecholamine concentrations and enhanced glycogenolysis. Endotoxin shock caused significant hypotension, acidosis, and impaired regional perfusion in all dogs. In addition, cardiac output, stroke volume, dP/dT, and left ventricular end-diastolic pressure fell and hyperglycemia and hyperinsulinemia occurred in all dogs after endotoxin injection.(ABSTRACT TRUNCATED AT 250 WORDS)

  5. An InDel in the Promoter ofAl-ACTIVATED MALATE TRANSPORTER9Selected during Tomato Domestication Determines Fruit Malate Contents and Aluminum Tolerance.

    Science.gov (United States)

    Ye, Jie; Wang, Xin; Hu, Tixu; Zhang, Fengxia; Wang, Bing; Li, Changxin; Yang, Tianxia; Li, Hanxia; Lu, Yongen; Giovannoni, James J; Zhang, Yuyang; Ye, Zhibiao

    2017-09-01

    Deciphering the mechanism of malate accumulation in plants would contribute to a greater understanding of plant chemistry, which has implications for improving flavor quality in crop species and enhancing human health benefits. However, the regulation of malate metabolism is poorly understood in crops such as tomato ( Solanum lycopersicum ). Here, we integrated a metabolite-based genome-wide association study with linkage mapping and gene functional studies to characterize the genetics of malate accumulation in a global collection of tomato accessions with broad genetic diversity. We report that TFM6 (tomato fruit malate 6), which corresponds to Al-ACTIVATED MALATE TRANSPORTER9 (Sl -ALMT9 in tomato), is the major quantitative trait locus responsible for variation in fruit malate accumulation among tomato genotypes. A 3-bp indel in the promoter region of Sl -ALMT9 was linked to high fruit malate content. Further analysis indicated that this indel disrupts a W-box binding site in the Sl -ALMT9 promoter, which prevents binding of the WRKY transcription repressor Sl-WRKY42, thereby alleviating the repression of Sl -ALMT9 expression and promoting high fruit malate accumulation. Evolutionary analysis revealed that this highly expressed Sl -ALMT9 allele was selected for during tomato domestication. Furthermore, vacuole membrane-localized Sl-ALMT9 increases in abundance following Al treatment, thereby elevating malate transport and enhancing Al resistance. © 2017 American Society of Plant Biologists. All rights reserved.

  6. 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

    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...... of leukocytes were nearly completely abolished after the apical supplementation of PC with CPBS, but not by CPBS alone. Ethanol up to 66 mM was not able to reverse this effect. A considerable part of the therapeutic and preventive effect of PC supplementation in ALD might result from a reduction of ethanol...

  7. Novel endotoxin-sequestering compounds with terephthalaldehyde-bis-guanylhydrazone scaffolds.

    Science.gov (United States)

    Khownium, Kriangsak; Wood, Stewart J; Miller, Kelly A; Balakrishna, Rajalakshmi; Nguyen, Thuan B; Kimbrell, Matthew R; Georg, Gunda I; David, Sunil A

    2006-03-01

    We have shown that lipopolyamines bind to the lipid A moiety of lipopolysaccharide, a constituent of Gram-negative bacterial membranes, and neutralize its toxicity in animal models of endotoxic shock. In an effort to identify non-polyamine scaffolds with similar endotoxin-recognizing features, we had observed an unusually high frequency of hits containing guanylhydrazone scaffolds in high-throughput screens. We now describe the syntheses and preliminary structure-activity relationships in a homologous series of bis-guanylhydrazone compounds decorated with hydrophobic functionalities. These first-generation compounds bind and neutralize lipopolysaccharide with a potency comparable to that of polymyxin B, a peptide antibiotic known to sequester LPS.

  8. Exposure to airborne microorganisms, dust and endotoxin during processing of peppermint and chamomile herbs on farms.

    Science.gov (United States)

    Skórska, Czesława; Sitkowska, Jolanta; Krysińska-Traczyk, Ewa; Cholewa, Grazyna; Dutkiewicz, Jacek

    2005-01-01

    The aim of this study was to determine the levels of microorganisms, dust and endotoxin in the air during processing of peppermint (Mentha piperita) and chamomile (Matricaria recutita) by herb farmers, and to examine the species composition of airborne microflora. Air samples were collected on glass fibre filters by use of personal samplers on 13 farms owned by herb cultivating farmers, located in Lublin province (eastern Poland). The concentrations of total viable microorganisms (bacteria + fungi) in the farm air during processing of peppermint herb were large, within a range from 895.1-6,015.8 x 10(3) cfu/m(3) (median 1,055.3 x 10(3) cfu/m(3)). During processing of chamomile herb they were much lower and varied within a range from 0.88-295.6 x 10(3) cfu/m(3) (median 27.3 x 10(3) cfu/m(3)). Gram-negative bacteria distinctly prevailed during processing of peppermint leaves, forming 46.4-88.5 % of the total airborne microflora. During processing of chamomile herb, Gram-negative bacteria were dominant at 3 out of 6 sampling sites forming 54.7-75.3 % of total microflora, whereas at the remaining 3 sites the most common were fungi forming 46.2-99.9 % of the total count. The species Pantoea agglomerans (synonyms: Erwinia herbicola, Enterobacter agglomerans ), having strong allergenic and endotoxic properties, distinctly prevailed among Gram-negative isolates. Among fungi, the most common species was Alternaria alternata. The concentrations of airborne dust and endotoxin determined on the examined herb farms were large. The concentrations of airborne dust during peppermint and chamomile processing ranged from 86.7-958.9 mg/m(3), and from 1.1-499.2 mg/m(3), respectively (medians 552.3 mg/m(3) and 12.3 mg/m(3)). The concentrations of airborne endotoxin determined during peppermint and chamomile processing were within a wide range 1.53-208.33 microg/m(3) and 0.005-2604.19 microg/m(3) respectively (medians 57.3 microg/m(3) and 0.96 microg/m(3)). In conclusion, farmers

  9. High exposure to endotoxin in farming is associated with less new-onset pollen sensitisation

    DEFF Research Database (Denmark)

    Elholm, Grethe; Schlünssen, Vivi; Doekes, Gert

    2018-01-01

    of 1113 young Danish farmers and rural controls, with a mean age of 19 years at baseline. Sensitisation to birch pollen, grass pollen, cat dander and house dust mite was measured by specific IgE levels in serum samples from baseline and at 15 years' follow-up. Changes in sensitisation were analysed...... was not associated with significant changes in cat dander and house dust mite sensitisation. CONCLUSIONS: High exposure to endotoxin during young adulthood appears to protect against new onset of pollen sensitisation, independent of childhood farm exposure....

  10. Exercise and IL-6 infusion inhibit endotoxin-induced TNF-alpha production in humans

    DEFF Research Database (Denmark)

    Starkie, Rebecca; Ostrowski, Sisse Rye; Jauffred, Sune

    2003-01-01

    and atherosclerosis. To test this hypothesis, we performed three experiments in which eight healthy males either rested (CON), rode a bicycle for 3 h (EX), or were infused with recombinant human IL-6 (rhIL-6) for 3 h while they rested. After 2.5 h, the volunteers received a bolus of Escherichia coli...... exercise and rhIL-6 infusion at physiological concentrations inhibit endotoxin-induced TNF-alpha production in humans. Hence, these data provide the first experimental evidence that physical activity mediates antiinflammatory activity and suggest that the mechanism include IL-6, which is produced...

  11. 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.

  12. 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...... in a human in vivo model of acute systemic low-grade inflammation, we measured circulating inflammatory mediators after intravenous administration of Escherichia coli endotoxin (LPS) bolus injection (0.1 ng/kg of body weight) in young healthy male subjects. The subjects were divided into three groups...

  13. The Effects of Ex Vivo Administration of Granulocyte-Macrophage Colony-Stimulating Factor and Endotoxin on Cytokine Release of Whole Blood Are Determined by Priming Conditions

    Directory of Open Access Journals (Sweden)

    A. Nierhaus

    2017-01-01

    Full Text Available Background. Lipopolysaccharide- (LPS- induced tumour necrosis factor alpha (TNFα secretion in critically ill patients can be considered as a measure of immune responsiveness. It can be enhanced by granulocyte-macrophage colony stimulating factor (GM-CSF. We investigated the effect of GM-CSF on ex vivo stimulated cytokine production using various preincubation regimens in healthy donors and patients with sepsis. Results. The maxima for the stimuli occurred 3 hours after stimulation. In donors, there was an increase (p<0.001 of LPS-induced TNFα levels following incubation with GM-CSF. The simultaneous incubation with GM-CSF and LPS caused an inhibition of TNFα production (p<0.001. Postincubation with GM-CSF did not yield any difference. In patients, preincubation with GM-CSF yielded an enhanced ex vivo TNFα-response when TNFα levels were low. Patients with increased TNFα concentrations did not show a GM-CSF stimulation effect. The GM-CSF preincubation yielded an increase of IL-8 production in patients and donors. Conclusions. This study demonstrates the immune-modulating properties of GM-CSF depending on the absence or presence of LPS or systemic TNFα. The timing of GM-CSF administration may be relevant for the modulation of the immune system in sepsis. The lack of stimulation in patients with high TNFα may represent endotoxin tolerance.

  14. Enhancement of intrinsic antitumor activity in spore-endotoxin mixtures of Bacillus thuringiensis by exposure to ultraviolet radiation

    International Nuclear Information System (INIS)

    Zamola, B.; Karminski-Zamola, G.; Fuks, Z.; Kubovic, M.; Wrishcer, M.

    1985-01-01

    Irradiation of spore-endotoxin mixtures from Bacillus thuringiensis cultures at 254 nm (60 μW cm -2 ) enhances their intrinsic antitumor potency as well as that of either component. The extent of enhancement depends on the length of exposure (optimum: 35 min) and may thus be due to photochemical changes of the endotoxin protein or/and to photoproduction of additional compounds with antitumor activity. Antitumor effects, expressed as survival rates of C57BL/6 mice inoculated with Lewis' mouse lung carcinoma and subjected to treatments 24 h later, depended on the number of doses of preparations administered (mixture, separated components). (author)

  15. 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...... exposed at ZT15. Taken together, this study shows that several key components of humoral immunity in tilapia exhibit circadian rhythms and adapt to photoperiodic changes. Further, results of the bacterial endotoxin challenge suggest that responsiveness of serum humoral factors to a biological insult...

  16. Dust, endotoxin, fungi, and bacteria exposure as determined by work task, season, and type of plant in a flower greenhouse

    DEFF Research Database (Denmark)

    Thilsing, T.; Madsen, A. M.; Basinas, I.

    2015-01-01

    ). Exposure to dust, endotoxin, fungi, and bacteria differed between seasons. Packing Lavandula, sticking, potting, and grading Rhipsalideae, and all examined tasks related to Campanula production except sticking increased dust exposure. Endotoxin exposure was increased during sticking Campanula and pinching...... exposure limit of 90 EU m(-3) was exceeded in 30% of the samples, which may have health implications for the employees. Exposure levels were found to vary depending on the tasks performed, and thereby results can be used to direct task-based initiatives to reduce workplace exposures....

  17. Effects of hydrogen-rich saline on endotoxin-induced uveitis

    Directory of Open Access Journals (Sweden)

    Wei-ming Yan

    2017-01-01

    Full Text Available The therapeutic effects of hydrogen-rich saline (HRS have been reported for a wide range of diseases mainly via selectively reducing the amount of reactive oxygen species. Oxidative stress plays an important role in the pathogenesis of uveitis and endotoxin-induced uveitis (EIU. In this study, we investigated whether HRS can mitigate EIU in rats. Sprague-Dawley rats were randomly divided into Norm group, Model group, HRS group, dexamethasone (DEX group, and rats in the latter three groups were injected with equal amount of lipopolysaccharide (LPS to induce EIU of different severities (by 1 mg/kg of LPS, or 1/8 mg/kg of LPS. Rats in HRS group were injected with HRS intraperitoneally at three different modes to purse an ameliorating effect of EIU (10 mL/kg of HRS immediately after injection of 1 mg/kg of LPS, 20 mL/kg of HRS once a day for 1 week before injection of 1 mg/kg of LPS and at 0, 0.5, 1, 2, 6, 8, 12 hours after LPS administration, or 20 mL/kg of HRS once a day for 1 week before injection of 1/8 mg/kg of LPS, and at 0, 0.5, 1, 2, 6, 8, 12, 24 hours and once a day for 3 weeks after LPS administration. Rats of DEX group were injected with 1 mL/kg of DEX solution intraperitoneally immediately after LPS administration. Rats in Norm and Model groups did not receive any treatment. All rats were examined under slit lamp microscope and graded according to the clinical signs of uveitis. Electroretinogram, quantitative analysis of protein in aqueous humor (AqH and histological examination of iris and ciliary body were also carried out. Our results showed that HRS did not obviously ameliorate the signs of uveitis under slit lamp examination and the inflammatory cells infiltration around iris and cilliary body of EIU induced by 1 mg/kg or 1/8 mg/kg of LPS (P > 0.05, while DEX significantly reduced the inflammation reflected by the above two indicators (P 0.05, while DEX had an obvious therapeutic effect (P < 0.05. However, HRS exerted an inhibition

  18. Selection for Cry3Bb1 resistance in a genetically diverse population of nondiapausing Western Corn Rootworm (Coleoptera: Chrysomelidae)

    Science.gov (United States)

    Five short-diapause laboratory lines of western corn rootworm (Diabrotica virgifera virgifera) were selected for resistance to MON863, a variety of corn genetically modified with the Bacillus thuringiensis (Bt) transgene that expresses the Cry3Bb1 endotoxin. Three of the selecte...

  19. Teaching Tolerance? Associational Diversity and Tolerance Formation

    DEFF Research Database (Denmark)

    Rapp, Carolin; Freitag, Markus

    2015-01-01

    Tolerance is a basic democratic principle that helps civil societies cope with rising levels of diversity stemming from increased immigration and individualism. During the last decade the question of how tolerance may be fostered has dominated debates in public and academic spheres. In this article...

  20. Human alveolar macrophage responses to air pollution particulates are associated with insoluble components of coarse material, including particulate endotoxin.

    Science.gov (United States)

    Soukup, J M; Becker, S

    2001-02-15

    Inhalation of particulate matter in the ambient air has been shown to cause pulmonary morbidity and exacerbate asthma. Alveolar macrophage (AM) are essential for effective removal of inhaled particles and microbes in the lower airways. While some particles minimally effect AM function others inhibit antimicrobial activity or cause cytokine and growth factor production leading to inflammation and tissue remodeling. This study has investigated the effects of water soluble (s) and insoluble (is) components of Chapel Hill, North Carolina ambient particulate matter in the size ranges 0.1-2.5 microm (PM2.5) and 2.5-10 microm (PM10) diameter, on human AM IL-6, TNFalpha, and MCP-1 cytokine production and host defense mechanisms including phagocytosis and oxidant production. Cytokines were found to be induced by isPM10 to a much higher extent (>50-fold) than sPM10, which in turn stimulated production better than isPM2.5, while sPM2.5 was inactive. Previous studies have indicated that endotoxin (ETOX) is a component of sPM10 responsible for cytokine production. Here, it is shown that inhibition of isPM10-induced cytokine production was partially achieved with polymyxin B and LPS-binding protein (LBP), but not with a metal chelator, implicating ETOX as a cytokine-inducing moiety also in isPM10. In addition to inducing cytokines, exposure to isPM10, but not the other PM fractions, also inhibited phagocytosis and oxidant generation in response to yeast. This inhibition was ETOX independent. The decrease in host defenses may be the result of apoptosis in the AM population, which was also found to be specifically caused by isPM10. These results show that the functional capacity of AM is selectively modulated by insoluble components of coarse PM, including the biocontaminant ETOX.

  1. Analusis by 252Cf plasma desorption mass spectrometry of Bordetella pertussis endotoxin after nitrous deamination

    Science.gov (United States)

    Deprun, C.; Karibian, D.; Caroff, M.

    1993-07-01

    Endotoxic lipopolysaccharides (LPSs) are the major components of Gram-negative bacterial outer membrane. Like many amphipathic molecules, they pose problems of heterogeneity, purity, solubility, and aggregation. Nevertheless, PDMS has recently have been applied to unmodified endotoxins composed of LPS having uip to five sugar units in their saccharide chain. The B. Pertussis LPSs, most of which have a dodecasaccharide domain, ahve been analysed by classical methods and the masses of the separate lipid and saccharide domains determined after rupture of the bond linking them. However, the acid treatment employed for these and most chemical analyses can also modify structures in the vicinity of the bond. In order to investigate this biologically-important region, the endotoxin was treated to nitrous deamination, which shortens the saccharide chain to five sugars, but preserves the acid-labile region of the LPS. The PDM spectrum of this derivative, which required new conditions for its desorption, confirmed the structure analysis and demonstrated the presence of at least four molecular species.

  2. Intestinal endotoxins as co-factors of liver injury in obstructive jaundice.

    Science.gov (United States)

    Mentes, B B; Tatlicioglu, E; Akyol, G; Uluoglu, O; Sultan, N; Yilmaz, E; Celebi, M; Taneri, F; Ferahkose, Z

    1996-01-01

    The concept of endotoxin-mediated rather than direct liver injury in biliary obstruction was investigated using the experimental rat model of bile duct ligation (BDL) and small bowel bacterial overgrowth (SBBO). Small identical doses of intravenous endotoxin (bacterial LPS) caused a significantly more severe liver injury in rats with BDL, compared with sham-operated rats, suggesting the possible contribution of LPS in this type of liver damage. BDL was then combined with surgically created jejunal self-filling blind loops, which resulted in SBBO. Plasma LPS level increased significantly, and once again a more severe liver injury, determined by liver histology and serum gamma-glutamyl transpeptidase levels, was observed compared with the control group of rats with BDL+self-emptying blind loops. The data presented suggest that small amounts of exogenous LPS and/or the ordinarily innocous amounts of LPS constantly absorbed from the intestinal tract may be critical in the hepatic damage caused by obstruction of the biliary tract.

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

    Directory of Open Access Journals (Sweden)

    Banun Kusumawardani

    2012-09-01

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

    Science.gov (United States)

    Chen, Xi; Howe, Jörg; Andrä, Jörg; Rössle, Manfred; Richter, Walter; da Silva, Ana Paula Galvão; Krensky, Alan M; Clayberger, Carol; Brandenburg, Klaus

    2007-10-01

    To combat infections by Gram-negative bacteria, it is not only necessary to kill the bacteria but also to neutralize pathogenicity factors such as endotoxin (lipopolysaccharide, LPS). The development of antimicrobial peptides based on mammalian endotoxin-binding proteins is a promising tool in the fight against bacterial infections, and septic shock syndrome. Here, synthetic peptides derived from granulysin (Gra-pep) were investigated in microbiological and biophysical assays to understand their interaction with LPS. We analyzed the influence of the binding of Gra-pep on (1) the acyl chain melting of the hydrophobic moiety of LPS, lipid A, by Fourier-transform spectroscopy, (2) the aggregate structure of LPS by small-angle X-ray scattering and cryo-transmission electron microscopy, and 3) the enthalpy change by isothermal titration calorimetry. In addition, the influence of Gra-pep on the incorporation of LPS and LPS-LBP (lipopolysaccharide-binding protein) complexes into negatively charged liposomes was monitored. Our findings demonstrate a characteristic change in the aggregate structure of LPS into multilamellar stacks in the presence of Gra-pep, but little or no change of acyl chain fluidity. Neutralization of LPS by Gra-pep is not due to a scavenging effect in solution, but rather proceeds after incorporation into target membranes, suggesting a requisite membrane-bound step.

  4. Acute hemodynamic changes during lung recruitment in lavage and endotoxin-induced ALI.

    Science.gov (United States)

    Odenstedt, Helena; Aneman, Anders; Kárason, Sigurbergur; Stenqvist, Ola; Lundin, Stefan

    2005-01-01

    To assess acute cardiorespiratory effects of recruitment manoeuvres in experimental acute lung injury. Experimental study in animal models of acute lung injury. Experimental laboratory at a University Medical Centre. Ten pigs with bronchoalveolar lavage and eight pigs with endotoxin-induced ALI. Two kinds of recruitment manoeuvres during 1 min; a) vital capacity manoeuvres (ViCM) consisting in a sustained inflation at 30 cmH(2)O and 40 cmH(2)O; b) manoeuvres obtained during ongoing pressure-controlled ventilation (PCRM) with peak airway pressure 30 cmH(2)O, positive end-expiratory pressure (PEEP) 15 and peak airway pressure 40, PEEP 20. Recruitment manoeuvres were repeated after volume expansion (dextran 8 ml/kg). Oxygenation, mean arterial, and pulmonary artery pressures, aortic, mesenteric, and renal blood flow were monitored. Lower pressure recruitment manoeuvres (ViCM30 and PCRM30/15) did not significantly improve oxygenation. With ViCM and PCRM at peak airway pressure 40 cmH(2)O, PaO(2) increased to similar levels in both lavage and endotoxin groups. Aortic blood flow was reduced from baseline during PCRM40/20 and ViCM40 by 57+/-3% and 61+/-6% in the lavage group and by 57+/-8% and 82+/-7% (Pdepression may be attenuated using recruitment manoeuvres during ongoing pressure-controlled ventilation and by prior volume expansion.

  5. Chronic lymphocytic leukemia: a paradigm of innate immune cross-tolerance.

    Science.gov (United States)

    Jurado-Camino, Teresa; Córdoba, Raúl; Esteban-Burgos, Laura; Hernández-Jiménez, Enrique; Toledano, Victor; Hernandez-Rivas, Jose-Angel; Ruiz-Sainz, Elena; Cobo, Teresa; Siliceo, María; Perez de Diego, Rebeca; Belda, Cristobal; Cubillos-Zapata, Carolina; López-Collazo, Eduardo

    2015-01-15

    Infections are a significant cause of morbidity and mortality in patients with chronic lymphocytic leukemia (CLL). The pathogenesis of infections is multifactorial and includes hypogammaglobulinemia, conventional therapy with alkylating drugs, and recently, purine analogs and mAb-associated T cells. Patients without these risk factors also suffer from infections, although the mechanism remains unknown. In a cohort of 70 patients with CLL, we demonstrated that their monocytes were locked into a refractory state and were unable to mount a classic inflammatory response to pathogens. In addition, they exhibited the primary features of endotoxin tolerance, including low cytokine production, high phagocytic activity, and impaired Ag presentation. The involvement of miR-146a in this phenomenon was suspected. We found miR-146a target genes, such as IRAK1 and TRAF6, were manifestly downregulated. Our study provides a new explanation for infections in patients with CLL and describes a cross-tolerance between endotoxins and tumors. Copyright © 2015 by The American Association of Immunologists, Inc.

  6. Lactose tolerance tests

    Science.gov (United States)

    Hydrogen breath test for lactose tolerance ... Two common methods include: Lactose tolerance blood test Hydrogen breath test The hydrogen breath test is the preferred method. It measures the amount of hydrogen ...

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

    Science.gov (United States)

    Norbäck, Dan; Hashim, Jamal Hisham; Markowicz, Pawel; Cai, Gui-Hong; Hashim, Zailina; Ali, Faridah; Larsson, Lennart

    2016-03-01

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

  8. Structure and function of cerebral and mesenteric resistance arteries in low-dose endotoxin-infused pregnant rats

    NARCIS (Netherlands)

    Wiegman, Marjon J; Van der Graaf, Anne Marijn; Henning, Robert H.; Zeeman, Gerda G.; Buikema, Hendrik; Faas, Marijke M.

    Objective: Since the cerebrovasculature likely plays a prominent role in the pathophysiology of eclampsia, we assessed the effects of low-dose endotoxin-induced experimental preeclampsia on the function and structure of rat posterior cerebral arteries (PCA) and mesenteric arteries (MA). Methods:

  9. A hybrid Bacillus thuringiensis delta-endotoxin gene gives resistance against a coleopteran and a lepidopteran pest in transgenic potato

    NARCIS (Netherlands)

    Naimov, S.; Dukiandjiev, S.; Maagd, de R.A.

    2003-01-01

    Expression of Bacillus thuringiensis delta-endotoxins has proven to be a successful strategy for obtaining insect resistance in transgenic plants. Drawbacks of expression of a single resistance gene are the limited target spectrum and the potential for rapid adaptation of the pest. Hybrid toxins

  10. Multiple linear regression and artificial neural networks for delta-endotoxin and protease yields modelling of Bacillus thuringiensis.

    Science.gov (United States)

    Ennouri, Karim; Ben Ayed, Rayda; Triki, Mohamed Ali; Ottaviani, Ennio; Mazzarello, Maura; Hertelli, Fathi; Zouari, Nabil

    2017-07-01

    The aim of the present work was to develop a model that supplies accurate predictions of the yields of delta-endotoxins and proteases produced by B. thuringiensis var. kurstaki HD-1. Using available medium ingredients as variables, a mathematical method, based on Plackett-Burman design (PB), was employed to analyze and compare data generated by the Bootstrap method and processed by multiple linear regressions (MLR) and artificial neural networks (ANN) including multilayer perceptron (MLP) and radial basis function (RBF) models. The predictive ability of these models was evaluated by comparison of output data through the determination of coefficient (R 2 ) and mean square error (MSE) values. The results demonstrate that the prediction of the yields of delta-endotoxin and protease was more accurate by ANN technique (87 and 89% for delta-endotoxin and protease determination coefficients, respectively) when compared with MLR method (73.1 and 77.2% for delta-endotoxin and protease determination coefficients, respectively), suggesting that the proposed ANNs, especially MLP, is a suitable new approach for determining yields of bacterial products that allow us to make more appropriate predictions in a shorter time and with less engineering effort.

  11. Characterization of the pH-Mediated solubility of Bacillus thuringiensis var. san diego native δ-endotoxin crystals

    Science.gov (United States)

    C. N. Koller; L. S. Bauer; R. M. Hollingworth

    1992-01-01

    Native crystals of Bacillus thuringiensis var. san diego, a coleopteran-specific δ-endotoxin, were metabolically labeled with [35S]methionine. Specific activity was 82,000 CPM/μg (2.44 Ci/mmol). Using a universal buffer formulated with the same ionic strength at every pH, we determined that...

  12. Endotoxin removal using 6,000 molecular weight cut-off polyacrylonitrile (PAN) and polysulfone (PS) hollow fiber ultrafilters.

    Science.gov (United States)

    Evans-Strickfaden, T T; Oshima, K H; Highsmith, A K; Ades, E W

    1996-01-01

    The removal of pyrogenic and microbial contaminants from high purity water and parental solutions during production is a concern to pharmaceutical manufacturers. In a previous study, 6000 molecular weight cut-off (MWCO) polyacrylonitrile (PAN) and polysulfone (PS) ultrafilters were shown to remove poliovirus (Titer reduction [Tr], > 6 logs) and phages T1 and PP7 (Tr, 7 logs) from ultrapure water, 0.85% saline with 1% trypicase soy broth, and Dulbecco's Eagle minimal essential medium with 10% fetal bovine serum (15). In this study, we evaluated the ability of the 6,000 MWCO PAN and PS ultrafilters to remove purified endotoxin (Escherichia coli lipopolysaccharide) which was added to commercial Water for Irrigation. An endotoxin concentration reduction of > 6 logs was achieved with both PAN and PS ultrafilters (detection sensitivity, 0.0031 EU/ml) when a minimum of 3.92 x 10(3) EU/ml of endotoxin was added to Water for Irrigation. These results indicate that the 6,000 MWCO PAN and PS ultrafilters are very effective in removing endotoxin from fluids such as Water for Irrigation.

  13. The immune response during the luteal phase of the ovarian cycle : increasing sensitivity of human monocytes to endotoxin

    NARCIS (Netherlands)

    Bouman, Annechien; Moes, H; Heineman, MJ; de Leij, LFMH; Faas, MM

    Objective: To test the hypothesis that during the luteal phase of the human ovarian cycle, as compared with the follicular phase, the percentage of cytokines producing peripheral monocytes after in vitro stimulation with endotoxin is increased. Design: Prospective study. Setting: Academic research

  14. Bacillus thuringiensis delta-endotoxin Cry1 hybrid proteins with increased activity against the Colorado potato beetle

    NARCIS (Netherlands)

    Naimov, S.; Weemen-Hendriks, M.; Dukiandjiev, S.; Maagd, de R.A.

    2001-01-01

    Cry1 delta-endotoxins of Bacillus thuringiensis are generally active against lepidopteran insects, but Cry1Ba and Cry1Ia have additional, though low, levels of activity against coleopterans such as the Colorado potato beetle. Here we report the construction of Cry1Ba/Cry1Ia hybrid toxins which have

  15. Oil composition of high-fat diet affects metabolic inflammation differently in connection with endotoxin receptors in mice.

    Science.gov (United States)

    Laugerette, Fabienne; Furet, Jean-Pierre; Debard, Cyrille; Daira, Patricia; Loizon, Emmanuelle; Géloën, Alain; Soulage, Christophe O; Simonet, Claire; Lefils-Lacourtablaise, Jennifer; Bernoud-Hubac, Nathalie; Bodennec, Jacques; Peretti, Noël; Vidal, Hubert; Michalski, Marie-Caroline

    2012-02-01

    Low-grade inflammation observed in obesity is a risk factor for cardiovascular disease. Recent studies revealed that this would be linked to gut-derived endotoxemia during fat digestion in high-fat diets, but nothing is known about the effect of lipid composition. The study was designed to test the impact of oil composition of high-fat diets on endotoxin metabolism and inflammation in mice. C57/Bl6 mice were fed for 8 wk with chow or isocaloric isolipidic diets enriched with oils differing in fatty acid composition: milk fat, palm oil, rapeseed oil, or sunflower oil. In vitro, adipocytes (3T3-L1) were stimulated or not with lipopolysaccharide (LPS; endotoxin) and incubated with different fatty acids. In mice, the palm group presented the highest level of IL-6 in plasma (P palm group was correlated with a greater ratio of LPS-binding protein (LBP)/sCD14 in plasma (P palm group, which was associated with lower inflammation in both plasma and adipose tissue despite higher plasma endotoxemia. Taken together, our results reveal that the palm oil-based diet resulted in the most active transport of LPS toward tissues via high LBP and low sCD14 and the greatest inflammatory outcomes. In contrast, a rapeseed oil-based diet seemed to result in an endotoxin metabolism driven toward less inflammatory pathways. This shows that dietary fat composition can contribute to modulate the onset of low-grade inflammation through the quality of endotoxin receptors.

  16. GLOMERULAR INFLAMMATION IN PREGNANT RATS AFTER INFUSION OF LOW-DOSE ENDOTOXIN - AN IMMUNOHISTOLOGICAL STUDY IN EXPERIMENTAL PREECLAMPSIA

    NARCIS (Netherlands)

    FAAS, MM; SCHUILING, GA; BALLER, JFW; BAKKER, WW

    1995-01-01

    Increased endotoxin sensitivity during pregnancy occurs in many animals, including rats. The mechanism of this phenomenon is not understood. In the present study it was investigated whether this increased sensitivity is reflected by an altered inflammatory pattern. Inflammatory cell influx, the

  17. 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...

  18. Single-use membrane adsorbers for endotoxin removal and purification of endogenous polysialic acid from Escherichia coli K1

    Directory of Open Access Journals (Sweden)

    Ingo de Vries

    2018-03-01

    After downstream processing the final product had neither detectable protein nor DNA contamination. Endotoxin content was below 3 EU mg−1. Investigation of the maximal chain length showed no effect of the harsh sodium hydroxide treatment during DSP on the stability of the polySia. Maximal chain length was ∼98 degree of polymerization.

  19. The Effects of the ND:YAG Laser on In vitro Fibroblast Attachment to Endotoxin Treated Root Surfaces

    Science.gov (United States)

    1991-05-01

    Preparation and Incubation .................................. 17 Scanning Electron Microscopic Observation 17 Data Collection and Analysis...fluorescence was seen regardless of the incubation time or endotoxin concentration. Brushing for one minute with a soft toothbrush removed most of the...However, the surface was brittle and fusion of enamel was not accomplished. Craters were produced at the point of laser impact . Peck and Peck (1967

  1. Relation of structure to function for the US reference standard endotoxin after exposure to 60Co radiation

    International Nuclear Information System (INIS)

    Csako, G.; Suba, E.A.; Ahlgren, A.; Tsai, C.M.; Elin, R.J.

    1986-01-01

    The structure and function of the highly purified US reference standard endotoxin (RSE) were studied after exposure to ionizing radiation from a 60 Co source. With increasing doses of radiation, the trilaminar ribbon-like structure of untreated endotoxin exhibited focal swelling, after which only spherical particles were seen by electron microscopy. These morphological changes were paralleled by the respective loss of O-side chain repeating units and pieces of the R-core from the lipopolysaccharide molecules, as demonstrated by electrophoresis. The biologic function of the irradiated endotoxin was assessed with a variety of tests. At higher doses of radiation, a direct relation was observed between the degradation of the molecular and supramolecular structure and the loss of biologic function. At lower doses of radiation, however, there was variability among the functional assays in their rate of change with progressive irradiation of the RSE. The results suggest that the carbohydrate moiety plays an important role both in determining the supramolecular structure and in modulating certain biologic activities of bacterial endotoxins

  2. Spatial and temporal variation in endotoxin and PM10 concentrations in ambient air in a livestock dense area

    NARCIS (Netherlands)

    de Rooij, Myrna; Heederik, Dick|info:eu-repo/dai/nl/072910542; Borlée, Floor|info:eu-repo/dai/nl/315138661; Hoek, Gerard|info:eu-repo/dai/nl/069553475; Wouters, Inge|info:eu-repo/dai/nl/274156652

    Several studies have reported associations between farming and respiratory health in neighboring residents. Health effects are possibly linked to fine dust and endotoxin emissions from livestock farms. Little is known about levels of these air pollutants in ambient air in livestock dense areas. We

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

    Science.gov (United States)

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

    2016-04-08

    Production of recombinant drugs in process-friendly endotoxin-free bacterial factories targets to a lessened complexity of the purification process combined with minimized biological hazards during product application. The development of nanostructured recombinant materials in innovative nanomedical activities expands such a need beyond plain functional polypeptides to complex protein assemblies. While Escherichia coli has been recently modified for the production of endotoxin-free proteins, no data has been so far recorded regarding how the system performs in the fabrication of smart nanostructured materials. We have here explored the nanoarchitecture and in vitro and in vivo functionalities of CXCR4-targeted, self-assembling protein nanoparticles intended for intracellular delivery of drugs and imaging agents in colorectal cancer. Interestingly, endotoxin-free materials exhibit a distinguishable architecture and altered size and target cell penetrability than counterparts produced in conventional E. coli strains. These variant nanoparticles show an eventual proper biodistribution and highly specific and exclusive accumulation in tumor upon administration in colorectal cancer mice models, indicating a convenient display and function of the tumor homing peptides and high particle stability under physiological conditions. The observations made here support the emerging endotoxin-free E. coli system as a robust protein material producer but are also indicative of a particular conformational status and organization of either building blocks or oligomers. This appears to be promoted by multifactorial stress-inducing conditions upon engineering of the E. coli cell envelope, which impacts on the protein quality control of the cell factory.

  4. Kinetics of intraocular tumor necrosis factor and interleukin-6 in endotoxin-induced uveitis in the rat

    NARCIS (Netherlands)

    de Vos, A. F.; van Haren, M. A.; Verhagen, C.; Hoekzema, R.; Kijlstra, A.

    1994-01-01

    To determine the kinetics of tumor necrosis factor (TNF) and interleukin-6 (IL-6) in serum and aqueous humor of rats with different susceptibilities to endotoxin-induced uveitis (EIU), after footpad injection of lipopolysaccharide (LPS). Samples were collected from EIU-susceptible Lewis rats and

  5. T-bet expression in the iris and spleen parallels disease expression during endotoxin-induced uveitis

    NARCIS (Netherlands)

    Li, B.; Yang, P.; Chu, L.; Zhou, H.; Huang, X.; Zhu, L.; Kijlstra, A.

    2007-01-01

    T lymphocytes have been implicated in the development of endotoxin-induced uveitis (EIU). T-bet is a Th1 cell-specific transcription factor that is involved in differentiation and effector functions. The aim of this study was to investigate kinetics of T-bet expression at the mRNA and protein levels

  6. Selection and characterisation of high ethanol tolerant ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-12-17

    Dec 17, 2008 ... Such sources include cashew, apple juice (Osho, 2005), palm wine (Bechem et al., 2007; Nwachukwu et al., 2006) and fermenting cassava tubers (Oyewole and Odunfa, 1988) among others. Despite the evolving trend of using bacteria for ethanol production, yeast is still the primary choice for fermentation ...

  7. Recognition and Toleration

    DEFF Research Database (Denmark)

    Lægaard, Sune

    2010-01-01

    Recognition and toleration are ways of relating to the diversity characteristic of multicultural societies. The article concerns the possible meanings of toleration and recognition, and the conflict that is often claimed to exist between these two approaches to diversity. Different forms...... or interpretations of recognition and toleration are considered, confusing and problematic uses of the terms are noted, and the compatibility of toleration and recognition is discussed. The article argues that there is a range of legitimate and importantly different conceptions of both toleration and recognition...

  8. Fault Tolerant Feedback Control

    DEFF Research Database (Denmark)

    Stoustrup, Jakob; Niemann, H.

    2001-01-01

    An architecture for fault tolerant feedback controllers based on the Youla parameterization is suggested. It is shown that the Youla parameterization will give a residual vector directly in connection with the fault diagnosis part of the fault tolerant feedback controller. It turns out...... that there is a separation be-tween the feedback controller and the fault tolerant part. The closed loop feedback properties are handled by the nominal feedback controller and the fault tolerant part is handled by the design of the Youla parameter. The design of the fault tolerant part will not affect the design...

  9. Selection and characterisation of an HD1-like Bacillus thuringiensis isolate with a high insecticidal activity against Spodoptera littoralis (Lepidoptera: Noctuidae).

    Science.gov (United States)

    Azzouz, Hichem; Kebaili-Ghribi, Jihene; ben Farhat-Touzri, Dalel; Daoud, Fatma; Fakhfakh, Ines; Tounsi, Slim; Jaoua, Samir

    2014-08-01

    Spodoptera littoralis (Boisduval) larvae are known by their susceptibility to Bacillus thuringiensis subsp. aizawai strains. In order to prevent the appearance of B. thuringiensis (Bt) resistance and to develop economical Bt-based biopesticides, the selection and the characterisation of a B. thuringiensis isolate toxic against S. littoralis larvae and overproducing δ-endotoxins were investigated. Among 124 Tunisian B. thuringiensis isolates assessed against S. littoralis larvae, four isolates showed toxicity similar to and higher than the toxicity of the aizawai strain HD133 and the kurstaki strain HD1 respectively. The plasmid pattern of the selected isolates was similar to that of HD1. Polymerase chain reaction (PCR) analysis using specific primers revealed that these isolates present different gene contents. The only detected gene encoding Spodoptera-specific toxin was cry9. The selected isolates were found to produce bipyramidal and cubic crystals. The assessment of δ-endotoxin production by these isolates showed that BUPM28 produced 43.71 and 80.81% more δ-endotoxin than HD1 and HD133 respectively. The application of osmotic or heat shock stress on the BUPM28 isolate made it possible to enhance δ-endotoxin production by 22 and 23% respectively. On the basis of its potent insecticidal activity and its high level of δ-endotoxin production, the BUPM28 isolate can be considered to be an effective alternative for the control of S. littoralis. © 2013 Society of Chemical Industry.

  10. Mechanical tolerance stackup and analysis

    CERN Document Server

    Fischer, Bryan R

    2004-01-01

    BackgroundDimensioning and TolerancingTolerance Format and Decimal PlacesConverting Plus/Minus Dimensions and Tolerances into Equal Bilaterally Toleranced DimensionsVariation and Sources of VariationTolerance AnalysisWorst-case Tolerance StackupsStatistical Tolerance StackupsGeometric Dimensioning and Tolerancing (GD&T)Converting Plus/Minus Tolerancing to Positional Tolerancing and Projected Tolerance ZonesDiametral and Radial Tolerance StackupsSpecifying Material Condition Modifiers and Their Effect on Tolerance Stackups The Tolerance Stackup SketchThe Tolerance Stackup Report FormTolerance S

  11. Salt tolerance in red clover (Trifolium pratense L.) seedlings

    African Journals Online (AJOL)

    Jane

    2011-08-15

    Aug 15, 2011 ... a result of evaporation and capillary rise of water. Selection of salt tolerant red clover genotypes would allow one to cultivate this crop on saline soils or with saline waters (Sidari et al., 2008). Germination and seedling characteristics are the most viable criteria used for selecting salt tolerance in plants.

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

    Directory of Open Access Journals (Sweden)

    GAO Baoxiu

    2014-01-01

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

  13. Tolerance Optimization for Mechanisms with Lubricated Joints

    International Nuclear Information System (INIS)

    Choi, J.-H.; Lee, S.J.; Choi, D.-H.

    1998-01-01

    This paper addresses an analytical approach to tolerance optimization for planar mechanisms with lubricated joints based on mechanical error analysis. The mobility method is applied to consider the lubrication effects at joints and planar mechanisms are stochastically defined by using the clearance vector model for mechanical error analysis. The uncertainties considered in the analysis are tolerances on link lengths and radial clearances and these are selected as design variables. To show the validity of the proposed method for mechanical error analysis, it is applied to two examples, and the results obtained are compared with those of Monte Carlo simulations. Based on the mechanical error analysis, tolerance optimizations are applied to the examples

  14. Transepithelial activation of human leukocytes by probiotics and commensal bacteria: Role of Enterobacteriaceae-type endotoxin

    DEFF Research Database (Denmark)

    Baeuerlein, Annette; Ackermann, Stefanie; Parlesak, Alexandr

    2009-01-01

    The goal of the current study was to clarify whether commercially available probiotics induce greater trans-epithelial activation of human leukocytes than do commensal, food-derived and pathogenic bacteria and to identify the compounds responsible for this activation. Eleven different bacterial...... strains, and some of their pathogen-associated molecular patterns, were incubated apically on a confluent layer of intestinal epithelial cells (Caco-2), which were basolaterally co-cultured with human mononuclear leukocytes. Only Gram-negative bacteria having Enterobacteriaceae-type endotoxin (commensal...... Escherichia coli K12, probiotic E. coli Nissle, EPEC) induced basolateral production of TNF-alpha, IFN-gamma, IL 6, 8, and 10. Gram-positive probiotics (Lactobacillus spp. and Bifidobacterium spp.) had virtually no effect. In addition, commensals (Enterococcus faecalis, Bacteroides vulgatus) and food...

  15. Transepithelial activation of human leukocytes by probiotics and commensal bacteria: role of Enterobacteriaceae-type endotoxin

    DEFF Research Database (Denmark)

    Bäuerlein, A.; Ackermann, S.; Parlesak, Alexandr

    2009-01-01

    The goal of the current study was to clarify whether commercially available probiotics induce greater trans-epithelial activation of human leukocytes than do commensal, food-derived and pathogenic bacteria and to identify the compounds responsible for this activation. Eleven different bacterial...... strains, and some of their pathogen-associated molecular patterns, were incubated apically on a confluent layer of intestinal epithelial cells (Caco-2), which were basolaterally co-cultured with human mononuclear leukocytes. Only Gram-negative bacteria having Enterobacteriaceae-type endotoxin (commensal...... Escherichia coli K12, probiotic E. coli Nissle, EPEC) induced basolateral production of TNF-alpha, IFN-gamma, IL 6, 8, and 10. Gram-positive probiotics (Lactobacillus spp. and Bifidobacterium spp.) had virtually no effect. In addition, commensals (Enterococcus faecalis, Bacteroides vulgatus) and food...

  16. 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.

  17. Diagnostic and prognostic significance of plasma endotoxin determination in febrile patients with haematological malignancies.

    Science.gov (United States)

    Yoshida, M; Obayashi, T; Tamura, H; Tanaka, S; Kawai, T; Sakamoto, S; Miura, Y

    1994-01-01

    We evaluated the clinical utility of a new endotoxin-specific chromogenic limulus test in febrile patients with haematological malignancies. The specificity is assured by the removal of factor G, which is sensitive to (1-->3)-beta-D-glucan, from horseshoe crab amoebocyte lysate. The sensitivity and specificity of the test to systemic gram-negative bacterial infections were 69.7 and 96.3%, respectively. Meanwhile, gram-negative bacteria grew in only 39.7% of endotoxaemic samples. Thus, it seems appropriate to consider gram-negative bacteraemia and endotoxaemia as different entities. Endotoxaemia was significantly associated with septic shock and infectious death, especially in patients with neutropenia. The new test, the results of which are available within 3 h, should help physicians to recognise this ominous sign early and to initiate a prompt countermeasure to endotoxaemia.

  18. A Meningococcal Outer Membrane Vesicle Vaccine Incorporating Genetically Attenuated Endotoxin Dissociates Inflammation From Immunogenicity

    Directory of Open Access Journals (Sweden)

    David J. Dowling

    2016-12-01

    Full Text Available Background. Group B Neisseria meningitidis, an endotoxin-producing gram-negative bacterium, causes the highest incidence of group B meningococcus (MenB disease in the first year of life. The Bexsero vaccine is indicated in Europe from 8 weeks of age. Endotoxin components of outer membrane vesicles (OMVs or soluble lipopolysaccharide (LPS represent a potential source of inflammation and residual reactogenicity. The purpose of this study was to compare novel candidate MenB vaccine formulations with licensed vaccines, including Bexsero, using age-specific in vitro culture systems.Methods. OMVs from wild type and inactivated lpxL1 gene mutant N. meningitidis strains were characterized in human neonatal and adult in vitro whole blood assays and dendritic cell arrays. OMVs were benchmarked against licensed vaccines, including Bexsero and whole cell pertussis formulations, with respect to Th-polarizing cytokine and PGE2 production, as well as cell surface activation markers (HLA-DR, CD86, CCR7. OMV immunogenicity was assessed in mice.Results. ΔlpxLI native OMVs demonstrated significantly less cytokine induction in human blood and DCs than Bexsero and most of the other pediatric vaccines (e.g., PedvaxHib, EasyFive, Bacillus Calmette–Guérin (BCG tested. Despite a much lower inflammatory profile in vitro than Bexsero, ΔlpxLI native OMVs still had moderate DC maturing ability and induced robust anti-N. meningitidis antibody responses after murine immunization.Conclusions. A meningococcal vaccine comprised of attenuated LPS-based OMVs with a limited inflammatory profile in vitro induces robust antigen-specific immunogenicity in vivo.

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

    Science.gov (United States)

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

    2012-11-01

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

  20. Synthetic anti-endotoxin peptides inhibit cytoplasmic LPS-mediated responses.

    Science.gov (United States)

    Pfalzgraff, Anja; Heinbockel, Lena; Su, Qi; Brandenburg, Klaus; Weindl, Günther

    2017-09-15

    Toll-like receptor (TLR) 4-independent recognition of lipopolysaccharide (LPS) in the cytosol by inflammatory caspases leads to non-canonical inflammasome activation and induction of IL-1 secretion and pyroptosis. The discovery of this novel mechanism has potential implications for the development of effective drugs to treat sepsis since LPS-mediated hyperactivation of caspases is critically involved in endotoxic shock. Previously, we demonstrated that Pep19-2.5, a synthetic anti-endotoxin peptide, efficiently neutralises pathogenicity factors of Gram-negative and Gram-positive bacteria and protects against sepsis in vivo. Here, we report that Pep19-2.5 inhibits the effects of cytoplasmic LPS in human myeloid cells and keratinocytes. In THP-1 monocytes and macrophages, the peptide strongly reduced secretion of IL-1β and LDH induced by intracellular LPS. In contrast, the TLR4 signaling inhibitor TAK-242 abrogates LPS-induced TNF and IL-1β secretion, but not pyroptotic cell death. Furthermore, Pep19-2.5 suppressed LPS-induced HMGB-1 production and caspase-1 activation in THP-1 monocytes. Consistent with this observation, we found impaired IL-1β and IL-1α release in LPS-stimulated primary monocytes in the presence of Pep19-2.5 and reduced LDH release and IL-1B and IL-1A expression in LPS-transfected HaCaT keratinocytes. Additionally, Pep19-2.5 completely abolished IL-1β release induced by LPS/ATP in macrophages via canonical inflammasome activation. In conclusion, we provide evidence that anti-endotoxin peptides inhibit the inflammasome/IL-1 axis induced by cytoplasmic LPS sensing in myeloid cells and keratinocytes and activation of the classical inflammasome by LPS/ATP which may contribute to the protection against bacterial sepsis and skin infections with intracellular Gram-negative bacteria. Copyright © 2017 Elsevier Inc. All rights reserved.

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

    Science.gov (United States)

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

    2004-05-01

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

  2. 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... thuringiensis var. kurstaki encapsulated in killed Pseudomonas fluorescens and the expression plasmid and...

  3. Susceptibilidade de espécies de plantas com potencial de fitorremediação do herbicida sulfentrazone Selection of plants tolerant to sulfentrazone and potential for phytoremediation

    Directory of Open Access Journals (Sweden)

    João Carlos Madalão

    2013-02-01

    species, and five doses of sulfentrazone (0, 200, 400, 800 and 1,600g ha-1. We evaluated the phytotoxicity of the herbicide, plant height and dry weight of shoots and roots. The species showing the least symptoms of phytotoxicity and the highest rates of height and fresh and dry biomass were C. juncea, C. ensiformis and D. lablab, which were selected as tolerant to the herbicide sulfentrazone and with potential for phytoremediation.

  4. Drought tolerance in potato (S. tuberosum L.): Can we learn from drought tolerance research in cereals?

    Science.gov (United States)

    Monneveux, Philippe; Ramírez, David A; Pino, María-Teresa

    2013-05-01

    Drought tolerance is a complex trait of increasing importance in potato. Our knowledge is summarized concerning drought tolerance and water use efficiency in this crop. We describe the effects of water restriction on physiological characteristics, examine the main traits involved, report the attempts to improve drought tolerance through in vitro screening and marker assisted selection, list the main genes involved and analyze the potential interest of native and wild potatoes to improve drought tolerance. Drought tolerance has received more attention in cereals than in potato. The review compares these crops for indirect selection methods available for assessment of drought tolerance related traits, use of genetic resources, progress in genomics, application of water saving techniques and availability of models to anticipate the effects of climate change on yield. It is concluded that drought tolerance improvement in potato could greatly benefit from the transfer of research achievements in cereals. Several promising research directions are presented, such as the use of fluorescence, reflectance, color and thermal imaging and stable isotope techniques to assess drought tolerance related traits, the application of the partial root-zone drying technique to improve efficiency of water supply and the exploitation of stressful memory to enhance hardiness. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

  5. Antimicrobial Tolerance in Biofilms

    OpenAIRE

    Stewart, Philip S.

    2015-01-01

    The tolerance of microorganisms in biofilms to antimicrobial agents is examined through a meta-analysis of literature data. A numerical tolerance factor comparing the rates of killing in the planktonic and biofilm states is defined to provide a quantitative basis for the analysis. Tolerance factors for biocides and antibiotics range over three orders of magnitude. This variation is not explained by taking into account the molecular weight of the agent, the chemistry of the agent, the substrat...

  6. Molecular leakage at implant-abutment connection--in vitro investigation of tightness of internal conical implant-abutment connections against endotoxin penetration.

    Science.gov (United States)

    Harder, Sönke; Dimaczek, Birka; Açil, Yaha; Terheyden, Hendrik; Freitag-Wolf, Sandra; Kern, Matthias

    2010-08-01

    Microleakage has been discussed as a major contributing factor for inflammatory reactions at the implant-abutment connection. In previous studies, the tightness against corpuscular bodies (viable bacteria) has been successfully investigated under static and dynamic conditions. The aim of this study was to investigate the tightness against endotoxins of two implant systems (AstraTech and Ankylos) with conical internal connections under static conditions. The inner parts of eight implants of each system were inoculated with endotoxin. Implants were screwed together with the respective abutments and stored under isostatic conditions in a supernatant of pyrogen-free water for 168 h. Supernatant samples were taken after 5 min, 24 h, 72 h, and 168 h, and endotoxin contamination was determined by the amebocyte-lysate test. Only one implant in the AstraTech group showed no sign of endotoxin contamination after 168 h, while the other implants showed contamination after varying storage times, respectively. The implants in the Ankylos group showed endotoxin contamination after only 5 min of storage in the supernatant solution. The tested internal conical implant-abutment connections appear to be unable to prevent endotoxin leakage. In average, Astra implants showed a higher tightness than Ankylos implants.

  7. Sex differences in the pro-inflammatory cytokine response to endotoxin unfold in vivo but not ex vivo in healthy humans.

    Science.gov (United States)

    Wegner, Alexander; Benson, Sven; Rebernik, Laura; Spreitzer, Ingo; Jäger, Marcus; Schedlowski, Manfred; Elsenbruch, Sigrid; Engler, Harald

    2017-07-01

    Clinical data indicate that inflammatory responses differ across sexes, but the mechanisms remain elusive. Herein, we assessed in vivo and ex vivo cytokine responses to bacterial endotoxin in healthy men and women to elucidate the role of systemic and cellular factors underlying sex differences in inflammatory responses. Participants received an i.v. injection of low-dose endotoxin (0.4 ng/kg body mass), and plasma TNF-α and IL-6 responses were analyzed over a period of 6 h. In parallel, ex vivo cytokine production was measured in endotoxin-stimulated blood samples obtained immediately before in vivo endotoxin administration. As glucocorticoids (GCs) play an important role in the negative feedback regulation of the inflammatory response, we additionally analyzed plasma cortisol concentrations and ex vivo GC sensitivity of cytokine production. Results revealed greater in vivo pro-inflammatory responses in women compared with men, with significantly higher increases in plasma TNF-α and IL-6 concentrations. In addition, the endotoxin-induced rise in plasma cortisol was more pronounced in women. In contrast, no sex differences in ex vivo cytokine production and GC sensitivity were observed. Together, these findings demonstrate major differences in in vivo and ex vivo responses to endotoxin and underscore the importance of systemic factors underlying sex differences in the inflammatory response.

  8. Tolerance: the forgotten child of plant resistance

    Directory of Open Access Journals (Sweden)

    Robert K.D. Peterson

    2017-10-01

    Full Text Available Plant resistance against insect herbivory has greatly focused on antibiosis, whereby the plant has a deleterious effect on the herbivore, and antixenosis, whereby the plant is able to direct the herbivore away from it. Although these two types of resistance may reduce injury and yield loss, they can produce selection pressures on insect herbivores that lead to pest resistance. Tolerance, on the other hand, is a more sustainable pest management strategy because it involves only a plant response and therefore does not cause evolution of resistance in target pest populations. Despite its attractive attributes, tolerance has been poorly studied and understood. In this critical, interpretive review, we discuss tolerance to insect herbivory and the biological and socioeconomic factors that have limited its use in plant resistance and integrated pest management. First, tolerance is difficult to identify, and the mechanisms conferring it are poorly understood. Second, the genetics of tolerance are mostly unknown. Third, several obstacles hinder the establishment of high-throughput phenotyping methods for large-scale screening of tolerance. Fourth, tolerance has received little attention from entomologists because, for most, their primary interest, research training, and funding opportunities are in mechanisms which affect pest biology, not plant biology. Fifth, the efforts of plant resistance are directed at controlling pest populations rather than managing plant stress. We conclude this paper by discussing future research and development activities.

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

    Science.gov (United States)

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

    2011-07-01

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

  10. The effectiveness of various biofiltration substrates in removing bacteria, endotoxins, and dust from ventilation system exhaust from a chicken hatchery.

    Science.gov (United States)

    Tymczyna, L; Chmielowiec-Korzeniowska, A; Drabik, A

    2007-10-01

    The objective of this study was to evaluate the effectiveness of various organic and organic-mineral biofilter media in purifying ventilation exhaust from a chicken hatchery room. Three different substrates were tested. Efficiency levels for the removal of dust, gram-negative bacteria, and bacterial endotoxin were recorded. The microbiological properties of the substrates were also studied. All of the biofilter substrates were highly effective in removing gram-negative bacteria, moderately effective in reducing dust levels, and only slightly effective in removing endotoxin. The substrate that was most efficient in retaining bioaerosols was the organic-mineral medium containing 20% halloysite, 40% compost, and 40% peat, which generally had at least satisfactory efficiency values for removing all of the contaminants tested.

  11. Compromise and Toleration

    DEFF Research Database (Denmark)

    Rostbøll, Christian F.

    Political compromise is akin to toleration, since both consist of an "agreement to disagree." Compromise and toleration also share a predicament of being regarded as ambiguous virtues that require of us to accept something we actually regard as wrong. However, we misunderstand the nature, justifi......Political compromise is akin to toleration, since both consist of an "agreement to disagree." Compromise and toleration also share a predicament of being regarded as ambiguous virtues that require of us to accept something we actually regard as wrong. However, we misunderstand the nature......, justification, and limits of compromise if we see it merely as a matter of toleration. While toleration is mainly a matter of accepting citizens' equal right to co-existence as subjects to law, political compromise includes the parties in making law – it makes them co-authors of law. Toleration entails....... The justification of compromise goes beyond, even if it includes, respect for each individual's right to determine her own conception of the good to an idea of respect for citizens as co-legislators. Second, the limits of compromise are not the same as the limits of toleration. The conditions for being included...

  12. Fault tolerant computing systems

    International Nuclear Information System (INIS)

    Randell, B.

    1981-01-01

    Fault tolerance involves the provision of strategies for error detection damage assessment, fault treatment and error recovery. A survey is given of the different sorts of strategies used in highly reliable computing systems, together with an outline of recent research on the problems of providing fault tolerance in parallel and distributed computing systems. (orig.)

  13. The Paradoxes of Tolerance

    Science.gov (United States)

    Pasamonik, Barbara

    2004-01-01

    Teachers who endeavor to build tolerant attitudes in their students often fall into the trap of political correctness. Political correctness can suspend free reflection on the differences inherent in otherness, which is the subject of tolerance, and creates an ideology of the generalized, abstract Other. As a result, teachers prefer to talk about…

  14. Remember Tolerance Differently

    DEFF Research Database (Denmark)

    Tønder, Lars

    2012-01-01

    This essay questions the linear conception of history which often accompanies the way contemporary democratic theory tends to disavow tolerance's discontinuities and remainders. In the spirit of Foucault's genealogy of descent, the idea is to develop a new sense of tolerance's history, not by inv...

  15. Microencapsulated Bifidobacterium longum subsp. infantis ATCC 15697 Favorably Modulates Gut Microbiota and Reduces Circulating Endotoxins in F344 Rats

    Directory of Open Access Journals (Sweden)

    Laetitia Rodes

    2014-01-01

    Full Text Available The gut microbiota is a bacterial bioreactor whose composition is an asset for human health. However, circulating gut microbiota derived endotoxins cause metabolic endotoxemia, promoting metabolic and liver diseases. This study investigates the potential of orally delivered microencapsulated Bifidobacterium infantis ATCC 15697 to modulate the gut microbiota and reduce endotoxemia in F344 rats. The rats were gavaged daily with saline or microencapsulated B. infantis ATCC 15697. Following 38 days of supplementation, the treated rats showed a significant (P < 0.05 increase in fecal Bifidobacteria (4.34 ± 0.46 versus 2.45 ± 0.25% of total and B. infantis (0.28 ± 0.21 versus 0.52 ± 0.12 % of total and a significant (P < 0.05 decrease in fecal Enterobacteriaceae (0.80 ± 0.45 versus 2.83 ± 0.63% of total compared to the saline control. In addition, supplementation with the probiotic formulation reduced fecal (10.52 ± 0.18 versus 11.29 ± 0.16 EU/mg; P = 0.01 and serum (0.33 ± 0.015 versus 0.30 ± 0.015 EU/mL; P = 0.25 endotoxins. Thus, microencapsulated B. infantis ATCC 15697 modulates the gut microbiota and reduces colonic and serum endotoxins. Future preclinical studies should investigate the potential of the novel probiotic formulation in metabolic and liver diseases.

  16. A study of the effect of Bacillus thuringiensis serotype H14 (subspecies israelensis) delta endotoxin on Musca larva.

    Science.gov (United States)

    Mehrabi, Mohammad Reza; Zoghimofrad, Laya; Mazinani, Mahboubeh; Akbarzadeh, Azim; Rahimi, Amirabbas

    2015-01-01

    House flies (Musca domestica) are of major public health concern in areas with poor sanitation and hygiene conditions. Biological control through the use of parasitoids and pathogens is one of the alternatives to the use of chemical pesticides for control of insects of public health importance. The effects of the delta endotoxin of Bacillus thuringiensis on house fly larval mortality were studied. Gel filtration and SDS-PAGE methods were used for separation and purification of proteins. Delta endotoxin was incubated with larvae in concentrations of 0.43 mg/mL and 0.27 mg/mL in bioassay tests. The results of this study indicated protein crystal toxicity against larvae of the house fly. A concentration of 0.43 mg/mL of this toxin caused 100% mortality in house fly larvae. The LD50 amount of these toxins was calculated as 125 µg/g. The results of this study suggest that the use of the protein crystal including delta endotoxin of Bacillus thuringiensis serotype H14 is an effective weapon in the biological fight against the house fly.

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

    Science.gov (United States)

    Zhou, Yan; Ruan, Zheng; Wen, Yanmei; Yang, Yuhui; Mi, Shumei; Zhou, Lili; Wu, Xin; Ding, Sheng; Deng, Zeyuan; Wu, Guoyao; Yin, Yulong

    2016-03-01

    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 chlorogenic acid-LD group, the area of visceral adipocyte was decreased and liver injury was ameliorated, as compared to LD group. In chlorogenic acid-LD group, serum triglycerides, free fatty acids, hepatic triglycerides and cholesterol were decreased, the proportion of C20:1, C24:1 and C18:3n-6, Δ9-18 and Δ6-desaturase activity index in the liver were decreased, and the proportion of C18:3n-3 acid was increased, compared to the LD group. Moreover, levels of phosphorylated AMP-activated protein kinase, carnitine palmitoyltransferase-I, and fatty acid β-oxidation were increased in chlorogenic acid-LD group compared to LD rats, whereas levels of fatty acid synthase and acetyl-CoA carboxylase were decreased. These findings demonstrate that chlorogenic acid effectively improves hepatic lipid dysregulation in rats by regulating fatty acid metabolism enzymes, stimulating AMP-activated protein kinase activation, and modulating levels of hepatic fatty acids.

  18. The effect of taurine on the relationship between NO, ADMA and homocysteine in endotoxin-mediated inflammation in HUVEC cultures.

    Science.gov (United States)

    Pasaoglu, Ozge Tugce; Turkozkan, Nurten; Ark, Mustafa; Polat, Belgin; Agilli, Mehmet; Yaman, Halil

    2014-10-01

    The aim of our study was to investigate the effect of taurine on the relationship between nitric oxide (NO), asymmetric dimethylarginine (ADMA) and homocysteine (Hcy) in endotoxin-induced human umblical vein endothelial cell (HUVEC) cultures. For this reason, four groups were formed (n=12). Control group consists of HUVEC cultures without any treatment. Lipopolysaccharide (LPS) and LPS+taurine groups were treated with 10 μg/mL endotoxin, 5 μg/mL taurine and endotoxin+taurine (same doses), respectively. Nitrite/nitrate (NOx), ADMA and Hcy levels were measured. There was a significant increase of NOx, ADMA and Hcy in endotoxemia (pTaurine treatment elevated NOx levels significantly (ptaurine and LPS + taurine group compared to control group, while it reduced NOx levels compared to LPS group. In contrast, taurine decreased ADMA levels to the control level both in taurine and taurine+LPS group compared to LPS. Hcy levels increased significantly compared to taurine group (pTaurine was effective on ADMA-NO relationship whereas no beneficial effect was observed in Hcy levels (p<0.05).

  19. Host defense peptides of thrombin modulate inflammation and coagulation in endotoxin-mediated shock and Pseudomonas aeruginosa sepsis.

    Directory of Open Access Journals (Sweden)

    Martina Kalle

    Full Text Available Gram-negative sepsis is accompanied by a disproportionate innate immune response and excessive coagulation mainly induced by endotoxins released from bacteria. Due to rising antibiotic resistance and current lack of other effective treatments there is an urgent need for new therapies. We here present a new treatment concept for sepsis and endotoxin-mediated shock, based on host defense peptides from the C-terminal part of human thrombin, found to have a broad and inhibitory effect on multiple sepsis pathologies. Thus, the peptides abrogate pro-inflammatory cytokine responses to endotoxin in vitro and in vivo. Furthermore, they interfere with coagulation by modulating contact activation and tissue factor-mediated clotting in vitro, leading to normalization of coagulation responses in vivo, a previously unknown function of host defense peptides. In a mouse model of Pseudomonas aeruginosa sepsis, the peptide GKY25, while mediating a modest antimicrobial effect, significantly inhibited the pro-inflammatory response, decreased fibrin deposition and leakage in the lungs, as well as reduced mortality. Taken together, the capacity of such thrombin-derived peptides to simultaneously modulate bacterial levels, pro-inflammatory responses, and coagulation, renders them attractive therapeutic candidates for the treatment of invasive infections and sepsis.

  20. Extreme flooding tolerance in Rorippa

    Science.gov (United States)

    Akman, Melis; Bhikharie, Amit V; Mustroph, Angelika; Sasidharan, Rashmi

    2014-01-01

    Low oxygen stress imposed by floods creates a strong selection force shaping plant ecosystems in flood-prone areas. Plants inhabiting these environments adopt various adaptations and survival strategies to cope with increasing water depths. Two Rorippa species, R. sylvestris and R. amphibia that grow in naturally flooded areas, have high submergence tolerance achieved by the so-called quiescence and escape strategies, respectively. In order to dissect the molecular mechanisms involved in these strategies, we investigated submergence-induced changes in gene expression in flooded roots of Rorippa species. There was a higher induction of glycolysis and fermentation genes and faster carbohydrate reduction in R. amphibia, indicating a higher demand for energy potentially leading to faster mortality by starvation. Moreover, R. sylvestris showed induction of genes improving submergence tolerance, potentially enhancing survival in prolonged floods. Additionally, we compared transcript profiles of these 2 tolerant species to relatively intolerant Arabidopsis and found that only Rorippa species induced various inorganic pyrophosphate dependent genes, alternatives to ATP demanding pathways, thereby conserving energy, and potentially explaining the difference in flooding survival between Rorippa and Arabidopsis. PMID:24525961

  1. 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

  2. Endotoxin-free purification for the isolation of Bovine Viral Diarrhoea Virus E2 protein from insoluble inclusion body aggregates

    Directory of Open Access Journals (Sweden)

    Mahony Timothy J

    2011-07-01

    Full Text Available Abstract Background Protein expression in Escherichia coli may result in the recombinant protein being expressed as insoluble inclusion bodies. In addition, proteins purified from E. coli contain endotoxins which need to be removed for in vivo applications. The structural protein, E2, from Bovine Viral Diarrhoea Virus (BVDV is a major immunogenic determinant, and is an ideal candidate as a subunit vaccine. The E2 protein contains 17 cysteine residues creating difficulties in E. coli expression. In this report we outline a procedure for successfully producing soluble and endotoxin-free BVDV E2 protein from inclusion bodies (IB. Results The expression of a truncated form of BVDV-E2 protein (E2-T1 in E. coli resulted in predominantly aggregated insoluble IB. Solubilisation of E2-T1 with high purity and stability from IB aggregates was achieved using a strong reducing buffer containing 100 mM Dithiothreitol. Refolding by dialysis into 50 mM Tris (pH 7.0 containing 0.2% Igepal CA630 resulted in a soluble but aggregated protein solution. The novel application of a two-phase extraction of inclusion body preparations with Triton X-114 reduced endotoxin in solubilised E2-T1 to levels suitable for in vivo use without affecting protein yields. Dynamic light scattering analyses showed 37.5% of the protein was monomeric, the remaining comprised of soluble aggregates. Mice immunised with E2-T1 developed a high titre antibody response by ELISA. Western hybridisation analysis showed E2-T1 was recognised by sera from immunised mice and also by several BVDV-E2 polyclonal and monoclonal antibodies. Conclusion We have developed a procedure using E. coli to produce soluble E2-T1 protein from IB, and due to their insoluble nature we utilised a novel approach using Triton X-114 to efficiently remove endotoxin. The resultant protein is immunogenic and detectable by BVDV-E2 specific antibodies indicating its usefulness for diagnostic applications and as a subunit

  3. Simple Screening Methods for Drought and Heat Tolerance in Cowpea

    International Nuclear Information System (INIS)

    Singh, B. B.

    2000-10-01

    Success in breeding for drought tolerance has not been as pronounced as for other traits. This is partly due to lack of simple, cheap and reliable screening methods to select drought tolerant plants/progenies from the segregating populations and partly due to complexity of factors involved in drought tolerance. Measuring drought tolerance through physiological parameters is expensive, time consuming and difficult to use for screening large numbers of lines and segregating populations. Since several factors/mechanisms (in shoot and root) operate independently and/or jointly to enable plants to cope with drought stress, drought tolerance appears as a complex trait. However, if these factors/mechanisms can be separated and studied individually, the components leading to drought tolerance will appear less complex and may be easy to manipulate by breeders. We have developed a simple box screening method for shoot drought tolerance in cowpea, which eliminates the effects of roots and permits non-destructive visual identification of shoot dehydration tolerance. We have also developed a 'root-box pin-board' method to study two dimensional root architecture of individual plants. Using these methods, we have identified two mechanisms of shoot drought tolerance in cowpea which are controlled by single dominant genes and major difference for root architecture among cowpea varieties. Combining deep and dense root system with shoot dehydration tolerance results into highly drought tolerant plants

  4. Tolerance of Four Tropical Tree Species to Heavy Petroleum Contamination

    NARCIS (Netherlands)

    Perez-Hernandez, I.; Ochoa-Gaona, S.; Schroeder, R.H.A.; Rivera-Cruz, M.C.; Geissen, V.

    2013-01-01

    Four species of trees were selected to evaluate the tolerance to heavy crude oil contamination by means of a tolerance index integrating germination, height, biomass and survival as variables. Fresh seeds to Cedrela odorata (tropical cedar), Haematoxylum campechianum (tinto bush), Swietenia

  5. Classification and salt tolerance analysis of barley varieties

    NARCIS (Netherlands)

    Katerji, N.; Hoorn, van J.W.; Hamdy, A.; Mastrorilli, M.; Fares, C.; Ceccarelli, S.; Grando, S.; Oweis, T.

    2006-01-01

    Six varieties of barley (Hordeum vulgare), five of which were provided by ICARDA, were tested in a green house experiment for their salt tolerance. Afterwards the ICARDA variety Melusine, selected from this experiment for its combination of high yield and salt tolerance, was compared in a lysimeter

  6. The prevalence of diabetes mellitus and impaired glucose tolerance ...

    African Journals Online (AJOL)

    The prevalence of diabetes mellitus and impaired glucose tolerance (IGT) was determined in 479 urbanised South African blacks (141 men and 338 women) of Zulu descent selected by cluster sampling in a suburb of Durban. All subjects underwent a modified glucose tolerance test whereby fasting and 2-hour postglucose ...

  7. Evaluation of drought tolerance indices among some winter ...

    African Journals Online (AJOL)

    Administrator

    2011-09-14

    watering stress treatments, Irrigation “time- .... STI and GMP were studied for selecting drought-tolerant cultivars in winter rapeseed cultivars. To do so, the 3D scatter plot was used because it allowed drought-tolerant cultivars ...

  8. Evaluation of drought tolerance in different growth stages of maize ...

    African Journals Online (AJOL)

    In order to find the best drought tolerant inbred lines, experiment was performed at the Agricultural College of Islamic Azad University, Shoushtar Branch, Iran during ... Data analysis revealed that the MP, GMP and STI indices were the more accurate criteria for selection of drought tolerant and high yielding inbred lines.

  9. Toleration out of respect?

    DEFF Research Database (Denmark)

    Lægaard, Sune

    2014-01-01

    Under conditions of pluralism different cultures, interests or values can come into conflict, which raises the problem of how to secure peaceful co-existence. The idea of toleration historically emerged as an answer to this problem. Recently Rainer Forst has argued that toleration should not just...... at hand. The respect conception therefore seems to be at best a theoretical idea belonging in ideal-theory, not a useful practical solution to actual conflicts under conditions of pluralism....... that avoids various so-called ‘paradoxes of toleration’. The paper first examines whether Forst’s respect conception can be applied descriptively to distinguish between actual patterns of behaviour and classify different acts of toleration. Then the focus is shifted to toleration out of respect as a normative...

  10. E. coli Endotoxin Modulates the Expression of Sirtuin Proteins in PBMC in Humans

    Directory of Open Access Journals (Sweden)

    Angela Storka

    2013-01-01

    Full Text Available Background. Sirtuin (SIRT proteins are class I histone deacetylases displaying gene regulatory functions in inflammatory, cancer, and metabolic diseases. These SIRT actions involve the nuclear factor κB and its inhibitor IκB pathway. However, the regulation of SIRT in vivo is still unclear. Material and Methods. In a human endotoxemia model, 20 healthy male subjects received an intravenous bolus of 2 ng/kg body weight Escherichia coli endotoxin (LPS. SIRT expression was investigated in peripheral blood mononuclear cells (PBMC with qPCR and Western blot before and 3 hours, 6 hours, and 24 hours after LPS challenge. Additionally, SIRT regulation was studied in vitro in cultivated PBMC after incubation with 20 ng/mL LPS. Results. A downregulation by >40% of SIRT1 mRNA was detectable 3 hours after LPS and of SIRT3 mRNA 6 hours after LPS. SIRT3, IκBα, and IκB-β protein expressions were decreased 3 and 6 hours after LPS. SIRT2 mRNA or protein expression did not change following LPS. These findings were consistent in vitro and associated with augmented phosphorylation of IκB-β. Discussion. In this E. coli endotoxemia model, SIRT1 and SIRT3 mRNA expressions in PBMC in humans were reduced after LPS challenge. This suggests that SIRT may represent an inflammatory target protein in vivo.

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

    Directory of Open Access Journals (Sweden)

    Yung-Ray Hsu

    2016-01-01

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

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

    International Nuclear Information System (INIS)

    Zhou, Yan; Ruan, Zheng; Zhou, Lili; Shu, Xugang; Sun, Xiaohong; Mi, Shumei; Yang, Yuhui; Yin, Yulong

    2016-01-01

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

  13. Cytotoxicity Analysis of Three Bacillus thuringiensis Subsp. israelensis δ-Endotoxins towards Insect and Mammalian Cells

    Science.gov (United States)

    Teixeira Corrêa, Roberto Franco; Ardisson-Araújo, Daniel Mendes Pereira; Monnerat, Rose Gomes; Ribeiro, Bergmann Morais

    2012-01-01

    Three members of the δ-endotoxin group of toxins expressed by Bacillus thuringiensis subsp. israelensis, Cyt2Ba, Cry4Aa and Cry11A, were individually expressed in recombinant acrystalliferous B. thuringiensis strains for in vitro evaluation of their toxic activities against insect and mammalian cell lines. Both Cry4Aa and Cry11A toxins, activated with either trypsin or Spodoptera frugiperda gastric juice (GJ), resulted in different cleavage patterns for the activated toxins as seen by SDS-PAGE. The GJ-processed proteins were not cytotoxic to insect cell cultures. On the other hand, the combination of the trypsin-activated Cry4Aa and Cry11A toxins yielded the highest levels of cytotoxicity to all insect cells tested. The combination of activated Cyt2Ba and Cry11A also showed higher toxic activity than that of toxins activated individually. When activated Cry4Aa, Cry11A and Cyt2Ba were used simultaneously in the same assay a decrease in toxic activity was observed in all insect cells tested. No toxic effect was observed for the trypsin-activated Cry toxins in mammalian cells, but activated Cyt2Ba was toxic to human breast cancer cells (MCF-7) when tested at 20 µg/mL. PMID:23029407

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

    Directory of Open Access Journals (Sweden)

    Carmine Vecchione

    2017-05-01

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

  15. Cytotoxicity analysis of three Bacillus thuringiensis subsp. israelensis δ-endotoxins towards insect and mammalian cells.

    Directory of Open Access Journals (Sweden)

    Roberto Franco Teixeira Corrêa

    Full Text Available Three members of the δ-endotoxin group of toxins expressed by Bacillus thuringiensis subsp. israelensis, Cyt2Ba, Cry4Aa and Cry11A, were individually expressed in recombinant acrystalliferous B. thuringiensis strains for in vitro evaluation of their toxic activities against insect and mammalian cell lines. Both Cry4Aa and Cry11A toxins, activated with either trypsin or Spodoptera frugiperda gastric juice (GJ, resulted in different cleavage patterns for the activated toxins as seen by SDS-PAGE. The GJ-processed proteins were not cytotoxic to insect cell cultures. On the other hand, the combination of the trypsin-activated Cry4Aa and Cry11A toxins yielded the highest levels of cytotoxicity to all insect cells tested. The combination of activated Cyt2Ba and Cry11A also showed higher toxic activity than that of toxins activated individually. When activated Cry4Aa, Cry11A and Cyt2Ba were used simultaneously in the same assay a decrease in toxic activity was observed in all insect cells tested. No toxic effect was observed for the trypsin-activated Cry toxins in mammalian cells, but activated Cyt2Ba was toxic to human breast cancer cells (MCF-7 when tested at 20 µg/mL.

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

    Science.gov (United States)

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

    2007-09-01

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

  17. Endotoxin-induced monocytic microparticles have contrasting effects on endothelial inflammatory responses.

    Science.gov (United States)

    Wen, Beryl; Combes, Valery; Bonhoure, Amandine; Weksler, Babette B; Couraud, Pierre-Olivier; Grau, Georges E R

    2014-01-01

    Septic shock is a severe disease state characterised by the body's life threatening response to infection. Complex interactions between endothelial cells and circulating monocytes are responsible for microvasculature dysfunction contributing to the pathogenesis of this syndrome. Here, we intended to determine whether microparticles derived from activated monocytes contribute towards inflammatory processes and notably vascular permeability. We found that endotoxin stimulation of human monocytes enhances the release of microparticles of varying phenotypes and mRNA contents. Elevated numbers of LPS-induced monocytic microparticles (mMP) expressed CD54 and contained higher levels of transcripts for pro-inflammatory cytokines such as TNF, IL-6 and IL-8. Using a prothrombin time assay, a greater reduction in plasma coagulation time was observed with LPS-induced mMP than with non-stimulated mMP. Co-incubation of mMP with the human brain endothelial cell line hCMEC/D3 triggered their time-dependent uptake and significantly enhanced endothelial microparticle release. Unexpectedly, mMP also modified signalling pathways by diminishing pSrc (tyr416) expression and promoted endothelial monolayer tightness, as demonstrated by endothelial impedance and permeability assays. Altogether, these data strongly suggest that LPS-induced mMP have contrasting effects on the intercellular communication network and display a dual potential: enhanced pro-inflammatory and procoagulant properties, together with protective function of the endothelium.

  18. 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

    Sepsis is a serious health problem associated with a range of infectious diseases in animals and humans. Early events of this syndrome can be mimicked by experimental administration of lipopolysaccharides (LPS). Compared with mice, small ruminants and humans are highly sensitive to LPS, making go...... 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....... goats valuable in inflammatory models. We performed a longitudinal study in eight Norwegian dairy goats that received LPS (0.1 μg/kg, Escherichia coli O26:B6) intravenously. A control group of five goats received corresponding volumes of sterile saline. Clinical examinations were performed continuously...... with the acute phase response, type I interferon signaling, LPS cascade and apoptosis, in addition to cytokines and chemokines were targeted. Pro-inflammatory genes, such as IL1B, CCL3 and IL8, were significantly up-regulated. Interestingly, increased mRNA levels of seven interferon stimulated genes (ISGs) were...

  19. MMP-3 Deficiency Alleviates Endotoxin-Induced Acute Inflammation in the Posterior Eye Segment

    Directory of Open Access Journals (Sweden)

    Inge Van Hove

    2016-11-01

    Full Text Available Matrix metalloproteinase-3 (MMP-3 is known to mediate neuroinflammatory processes by activating microglia, disrupting blood–central nervous system barriers and supporting neutrophil influx into the brain. In addition, the posterior part of the eye, more specifically the retina, the retinal pigment epithelium (RPE and the blood–retinal barrier, is affected upon neuroinflammation, but a role for MMP-3 during ocular inflammation remains elusive. We investigated whether MMP-3 contributes to acute inflammation in the eye using the endotoxin-induced uveitis (EIU model. Systemic administration of lipopolysaccharide induced an increase in MMP-3 mRNA and protein expression level in the posterior part of the eye. MMP-3 deficiency or knockdown suppressed retinal leukocyte adhesion and leukocyte infiltration into the vitreous cavity in mice subjected to EIU. Moreover, retinal and RPE mRNA levels of intercellular adhesion molecule 1 (Icam1, interleukin 6 (Il6, cytokine-inducible nitrogen oxide synthase (Nos2 and tumor necrosis factor α (Tnfα, which are key molecules involved in EIU, were clearly reduced in MMP-3 deficient mice. In addition, loss of MMP-3 repressed the upregulation of the chemokines monocyte chemoattractant protein (MCP-1 and (C-X-C motif ligand 1 (CXCL1. These findings suggest a contribution of MMP-3 during EIU, and its potential use as a therapeutic drug target in reducing ocular inflammation.

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

    Directory of Open Access Journals (Sweden)

    Margaret T Armstrong

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