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Sample records for extracellular traps nets

  1. Neutrophil extracellular traps in dermatology: Caught in the NET.

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    Hoffmann, Jochen H O; Enk, Alexander H

    2016-10-01

    Neutrophil, or polymorphonuclear granulocytes (PMN) constitute the most abundant type of leucocytes in peripheral human blood. One of the major advances in the last decade was the discovery of neutrophil extracellular trap (NET) formation: a process by which neutrophils externalize web-like chromatin strands decorated with antimicrobial peptides. These structures were soon implicated in immune defense and auto-immunity alike and now link neutrophils to the pathogenesis of a variety of diseases of dermatological relevance. Currently, NET formation is mainly subdivided into suicidal and vital NETosis. Controversy exists regarding the capacity of NETs to kill pathogens, and little is known about the way NETs are formed in vivo. Here, we discuss the current terminology, methods for NET quantification, pathways leading to NET formation, and the role of NETs in systemic and cutaneous immune defense and auto-immunity, with a focus on psoriasis and systemic lupus erythematosus.

  2. Yersinia enterocolitica-mediated degradation of neutrophil extracellular traps (NETs).

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    Möllerherm, Helene; Neumann, Ariane; Schilcher, Katrin; Blodkamp, Stefanie; Zeitouni, Nathalie E; Dersch, Petra; Lüthje, Petra; Naim, Hassan Y; Zinkernagel, Annelies S; von Köckritz-Blickwede, Maren

    2015-12-01

    Neutrophil extracellular trap (NET) formation is described as a tool of the innate host defence to fight against invading pathogens. Fibre-like DNA structures associated with proteins such as histones, cell-specific enzymes and antimicrobial peptides are released, thereby entrapping invading pathogens. It has been reported that several bacteria are able to degrade NETs by nucleases and thus evade the NET-mediated entrapment. Here we studied the ability of three different Yersinia serotypes to induce and degrade NETs. We found that the common Yersinia enterocolitica serotypes O:3, O:8 and O:9 were able to induce NETs in human blood-derived neutrophils during the first hour of co-incubation. At later time points, the NET amount was reduced, suggesting that degradation of NETs has occurred. This was confirmed by NET degradation assays with phorbol-myristate-acetate-pre-stimulated neutrophils. In addition, we found that the Yersinia supernatants were able to degrade purified plasmid DNA. The absence of Ca(2+) and Mg(2+) ions, but not that of a protease inhibitor cocktail, completely abolished NET degradation. We therefore postulate that Y. enterocolitica produces Ca(2+)/Mg(2+)-dependent NET-degrading nucleases as shown for some Gram-positive pathogens.

  3. Neutrophil extracellular traps (NETs) and infection-related vascular dysfunction.

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    Gardiner, Elizabeth E; Andrews, Robert K

    2012-11-01

    The innate immune system orchestrated by leukocytes primarily neutrophils, serves to remove dead and dying host cells and to provide protection against invasion by pathogens. Failure of this system results in the onset of sepsis leading to grave consequences for the host. Together with mechanical methods to physically isolate and remove the pathogen, neutrophils also release an important set of proinflammatory biological modulators that mediate recruitment of additional cells to a site of infection and amplify the innate protective response. Additionally, neutrophils release highly charged mixtures of DNA and nuclear proteins named neutrophil extracellular traps (NETs). These electrostatically-charged adhesive networks trigger intrinsic coagulation, limit dispersion and entrap the pathogens. NETs also contain the neutrophil secretary granule-derived serine proteases, neutrophil elastase and cathepsin G, known to regulate the reactivity of both neutrophils and platelets. Since the characterization of NETs in 2004, new studies of their functional effect in vivo continue to expand upon unexpected extracellular roles for DNA, and in doing so renew attention to the haemostatic role of the leukocyte. This review will provide a basic description of NETs and examine current knowledge of this important system of defense, including recent work illustrating a role for NETs in activation of thrombosis. Copyright © 2012 Elsevier Ltd. All rights reserved.

  4. The effect of clindamycin and amoxicillin on neutrophil extracellular trap (NET) release.

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    Bystrzycka, Weronika; Moskalik, Aneta; Sieczkowska, Sandra; Manda-Handzlik, Aneta; Demkow, Urszula; Ciepiela, Olga

    2016-01-01

    Neutrophil extracellular traps (NETs) are threads of nuclear DNA complexed with antimicrobial proteins released by neutrophils to extracellular matrix to bind, immobilise, and kill different pathogens. NET formation is triggered by different physiological and non-physiological stimulants. It is also suggested that antibiotics could be non-physiological compounds that influence NET release. The aim of the study was to investigate the effect of clindamycin and amoxicillin on NET release and the phagocyte function of neutrophils. Neutrophils isolated from healthy donors by density centrifugation method were incubated with amoxicillin or clindamycin for two hours, and then NET release was stimulated with phorbol 12-myristate 13-acetate (PMA). After three hours of incubation with PMA NETs were quantified as amount of extracellular DNA by fluorometry and visualised by immunofluorescent microscopy. The percent of phagocyting cells was measured by flow cytometry. We showed that amoxicillin induces NET formation (increase of extracellular DNA fluorescence, p = 0.03), while clindamycin had no influence on NET release (p > 0.05), as confirmed by quantitative measurement and fluorescent microscopy. Regarding phagocyte function, both antibiotics increased bacterial uptake (43.3% and 61.6% median increase for amoxicillin and clindamycin, respectively). We concluded that the ability of antibiotics to modulate NET release depends on the antibiotic used and is not associated with their ability to influence phagocytosis.

  5. Neutrophil extracellular traps (NETs) and the role of platelets in infection.

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    Andrews, Robert K; Arthur, Jane F; Gardiner, Elizabeth E

    2014-10-01

    In addition to playing a central role in normal haemostasis, platelets make important contributions to host inflammatory and immune responses to injury or infection. Under pathophysiological conditions where platelet function is not tightly controlled, platelets also play critical roles in pathogenic processes underlying cardiovascular disease, uncontrolled inflammation, coagulopathy and in tumour metastasis. Neutrophil extracellular traps (NETs) are webs of histone-modified nuclear material extruded from activated neutrophils during inflammatory responses and these degranulation events can be directly triggered by platelet/neutrophil engagement. Emerging research describes how NETs influence platelet function, particularly in the setting of infection and inflammation. Especially intriguing is the potential for platelet-driven coagulation to be modulated by NETs in plasma and interstitial spaces. These findings also reveal new perspectives related to improved therapy for venous thrombosis.

  6. Automatic determination of NET (neutrophil extracellular traps) coverage in fluorescent microscopy images.

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    Coelho, Luis Pedro; Pato, Catarina; Friães, Ana; Neumann, Ariane; von Köckritz-Blickwede, Maren; Ramirez, Mário; Carriço, João André

    2015-07-15

    Neutrophil extracellular traps (NETs) are believed to be essential in controlling several bacterial pathogens. Quantification of NETs in vitro is an important tool in studies aiming to clarify the biological and chemical factors contributing to NET production, stabilization and degradation. This estimation can be performed on the basis of fluorescent microscopy images using appropriate labelings. In this context, it is desirable to automate the analysis to eliminate both the tedious process of manual annotation and possible operator-specific biases. We propose a framework for the automated determination of NET content, based on visually annotated images which are used to train a supervised machine-learning method. We derive several methods in this framework. The best results are obtained by combining these into a single prediction. The overall Q(2) of the combined method is 93%. By having two experts label part of the image set, we were able to compare the performance of the algorithms to the human interoperator variability. We find that the two operators exhibited a very high correlation on their overall assessment of the NET coverage area in the images (R(2) is 97%), although there were consistent differences in labeling at pixel level (Q(2), which unlike R(2) does not correct for additive and multiplicative biases, was only 89%). Open source software (under the MIT license) is available at https://github.com/luispedro/Coelho2015_NetsDetermination for both reproducibility and application to new data. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  7. Escaping Underground Nets: Extracellular DNases Degrade Plant Extracellular Traps and Contribute to Virulence of the Plant Pathogenic Bacterium Ralstonia solanacearum.

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    Tran, Tuan Minh; MacIntyre, April; Hawes, Martha; Allen, Caitilyn

    2016-06-01

    Plant root border cells have been recently recognized as an important physical defense against soil-borne pathogens. Root border cells produce an extracellular matrix of protein, polysaccharide and DNA that functions like animal neutrophil extracellular traps to immobilize pathogens. Exposing pea root border cells to the root-infecting bacterial wilt pathogen Ralstonia solanacearum triggered release of DNA-containing extracellular traps in a flagellin-dependent manner. These traps rapidly immobilized the pathogen and killed some cells, but most of the entangled bacteria eventually escaped. The R. solanacearum genome encodes two putative extracellular DNases (exDNases) that are expressed during pathogenesis, suggesting that these exDNases contribute to bacterial virulence by enabling the bacterium to degrade and escape root border cell traps. We tested this hypothesis with R. solanacearum deletion mutants lacking one or both of these nucleases, named NucA and NucB. Functional studies with purified proteins revealed that NucA and NucB are non-specific endonucleases and that NucA is membrane-associated and cation-dependent. Single ΔnucA and ΔnucB mutants and the ΔnucA/B double mutant all had reduced virulence on wilt-susceptible tomato plants in a naturalistic soil-soak inoculation assay. The ΔnucA/B mutant was out-competed by the wild-type strain in planta and was less able to stunt root growth or colonize plant stems. Further, the double nuclease mutant could not escape from root border cells in vitro and was defective in attachment to pea roots. Taken together, these results demonstrate that extracellular DNases are novel virulence factors that help R. solanacearum successfully overcome plant defenses to infect plant roots and cause bacterial wilt disease.

  8. Escaping Underground Nets: Extracellular DNases Degrade Plant Extracellular Traps and Contribute to Virulence of the Plant Pathogenic Bacterium Ralstonia solanacearum.

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    Tuan Minh Tran

    2016-06-01

    Full Text Available Plant root border cells have been recently recognized as an important physical defense against soil-borne pathogens. Root border cells produce an extracellular matrix of protein, polysaccharide and DNA that functions like animal neutrophil extracellular traps to immobilize pathogens. Exposing pea root border cells to the root-infecting bacterial wilt pathogen Ralstonia solanacearum triggered release of DNA-containing extracellular traps in a flagellin-dependent manner. These traps rapidly immobilized the pathogen and killed some cells, but most of the entangled bacteria eventually escaped. The R. solanacearum genome encodes two putative extracellular DNases (exDNases that are expressed during pathogenesis, suggesting that these exDNases contribute to bacterial virulence by enabling the bacterium to degrade and escape root border cell traps. We tested this hypothesis with R. solanacearum deletion mutants lacking one or both of these nucleases, named NucA and NucB. Functional studies with purified proteins revealed that NucA and NucB are non-specific endonucleases and that NucA is membrane-associated and cation-dependent. Single ΔnucA and ΔnucB mutants and the ΔnucA/B double mutant all had reduced virulence on wilt-susceptible tomato plants in a naturalistic soil-soak inoculation assay. The ΔnucA/B mutant was out-competed by the wild-type strain in planta and was less able to stunt root growth or colonize plant stems. Further, the double nuclease mutant could not escape from root border cells in vitro and was defective in attachment to pea roots. Taken together, these results demonstrate that extracellular DNases are novel virulence factors that help R. solanacearum successfully overcome plant defenses to infect plant roots and cause bacterial wilt disease.

  9. “The NET outcome”: are neutrophil extracellular traps of any relevance to the pathophysiology of autoimmune disorders in childhood?

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    Stavros Giaglis

    2016-09-01

    Full Text Available Neutrophil extracellular trap (NET formation represents a form of cell death distinct from apoptosis or necrosis, by which invading pathogens are simultaneously entangled and potentially eliminated. Increased NET formation is observed in systemic lupus erythematosus (SLE, rheumatoid arthritis (RA, antineutrophil cytoplasmic antibody (ANCA-associated and small vessel vasculitis (SVV, antiphospholipid antibody syndrome (APS and psoriasis. NETs contribute to the pathogenesis of autoimmunity by exposing cryptic autoepitopes, which may facilitate the generation of autoantibodies, induce the production of interferons and activate the complement cascade. In SLE, augmented disease activity and renal disease are associated with increased NET formation, so that NETs could serve as a marker for the monitoring of disease activity. NETs can additionally cause endothelial cell damage and death and stimulate inflammation in atheromatous plaques, adding to the accelerated atherosclerosis witnessed in autoimmune disease. Since NETs induce production of interferons, assessing the extent of NET formation might facilitate the prediction of IFN-alpha levels and identification of SLE patients with presumably better responses to anti-IFN-alpha therapies or other novel therapeutic concepts, such as N-acetyl-cysteine and inhibitors of DNase 1, and peptidylarginine deiminase 4 (PAD4, which also target NETs. In summary, the study of NETs provides a novel approach to the understanding of autoimmune disease pathogenesis and opens new vistas in the development of sensitive disease markers and therapies.

  10. “The NET Outcome”: Are Neutrophil Extracellular Traps of Any Relevance to the Pathophysiology of Autoimmune Disorders in Childhood?

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    Giaglis, Stavros; Hahn, Sinuhe; Hasler, Paul

    2016-01-01

    Neutrophil extracellular trap (NET) formation represents a form of cell death distinct from apoptosis or necrosis, by which invading pathogens are simultaneously entangled and potentially eliminated. Increased NET formation is observed in systemic lupus erythematosus (SLE), rheumatoid arthritis, antineutrophil cytoplasmic antibody-associated small vessel vasculitis, antiphospholipid antibody syndrome (APS), and psoriasis. NETs contribute to the pathogenesis of autoimmunity by exposing cryptic autoepitopes, which may facilitate the generation of autoantibodies, induce the production of interferons, and activate the complement cascade. In SLE, augmented disease activity and renal disease are associated with increased NET formation, so that NETs could serve as a marker for the monitoring of disease activity. NETs can additionally cause endothelial cell damage and death and stimulate inflammation in atheromatous plaques, adding to the accelerated atherosclerosis witnessed in autoimmune disease. Since NETs induce production of interferons, assessing the extent of NET formation might facilitate the prediction of IFN-alpha levels and identification of SLE patients with presumably better responses to anti-IFN-alpha therapies or other novel therapeutic concepts, such as N-acetyl-cysteine and inhibitors of DNase 1 and peptidylarginine deiminase 4 (PAD4), which also target NETs. In summary, the study of NETs provides a novel approach to the understanding of autoimmune disease pathogenesis in childhood and opens new vistas in the development of sensitive disease markers and targeted therapies. PMID:27679792

  11. Neutrophil Extracellular Traps Identification in Tegumentary Lesions of Patients with Paracoccidioidomycosis and Different Patterns of NETs Generation In Vitro.

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    Amanda Manoel Della Coletta

    Full Text Available Paracoccidioidomycosis (PCM is a systemic mycosis, endemic in most Latin American countries, especially in Brazil. It is caused by the thermo-dimorphic fungus of the genus Paracoccidioides (Paracoccidioides brasiliensis and Paracoccidioides lutzii. Innate immune response plays a crucial role in host defense against fungal infections, and neutrophils (PMNs are able to combat microorganisms with three different mechanisms: phagocytosis, secretion of granular proteins, which have antimicrobial properties, and the most recent described mechanism called NETosis. This new process is characterized by the release of net-like structures called Neutrophil Extracellular Traps (NETs, which is composed of nuclear (decondensed DNA and histones and granular material such as elastase. Several microorganisms have the ability of inducing NETs formation, including gram-positive and gram-negative bacteria, viruses and some fungi. We proposed to identify NETs in tegumentary lesions of patients with PCM and to analyze the interaction between two strains of P. brasiliensis and human PMNs by NETs formation in vitro. In this context, the presence of NETs in vivo was evidenced in tegumentary lesions of patients with PCM by confocal spectrum analyzer. Furthermore, we showed that the high virulent P. brasiliensis strain 18 (Pb18 and the lower virulent strain Pb265 are able to induce different patterns of NETs formation in vitro. The quantification of extracellular DNA corroborates the idea of the ability of P. brasiliensis in inducing NETs release. In conclusion, our data show for the first time the identification of NETs in lesions of patients with PCM and demonstrate distinct patterns of NETs in cultures challenged with fungi in vitro. The presence of NETs components both in vivo and in vitro open new possibilities for the detailed investigation of immunity in PCM.

  12. Selected mucolytic, anti-inflammatory and cardiovascular drugs change the ability of neutrophils to form extracellular traps (NETs).

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    Zawrotniak, Marcin; Kozik, Andrzej; Rapala-Kozik, Maria

    2015-01-01

    Neutrophils form the first line of host defense against infections that combat pathogens using two major mechanisms, the phagocytosis or the release of neutrophil extracellular traps (NETs). The netosis (NET formation) exerts additional, unfavorable effects on the fitness of host cells and is also involved at the sites of lung infection, increasing the mucus viscosity and in the circulatory system where it can influence the intravascular clot formation. Although molecular mechanisms underlying the netosis are still incompletely understood, a role of NADPH oxidase that activates the production of reactive oxygen species (ROS) during the initiation of NETs has been well documented. Since several commonly used drugs can affects the netosis, our current study was aimed to determine the effects of selected mucolytic, anti-inflammatory and cardiovascular drugs on NET formation, with a special emphasis on ROS production and NADPH oxidase activity. The treatment of neutrophils with N-acetylcysteine, ketoprofen and ethamsylate reduced the production of ROS by these cells in a dose-dependent manner. NET formation was also modulated by selected drugs. N-acetylcysteine inhibited the netosis but in the presence of H2O2 this neutrophil ability was restored, indicating that N-acetylcysteine may influence the NET formation by modulating ROS productivity. The administration of ethamsylate led to a significant reduction in NET formation and this effect was not restored by H2O2 or S. aureus, suggesting the unexpected additional side effects of this drug. Ketoprofen seemed to promote ROS-independent NET release, simultaneously inhibiting ROS production. The results, obtained in this study strongly suggest that the therapeutic strategies applied in many neutrophil-mediated diseases should take into account the NET-associated effects.

  13. At the Bench: Neutrophil extracellular traps (NETs) highlight novel aspects of innate immune system involvement in autoimmune diseases.

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    Grayson, Peter C; Kaplan, Mariana J

    2016-02-01

    The putative role of neutrophils in host defense against pathogens is a well-recognized aspect of neutrophil function. The discovery of neutrophil extracellular traps has expanded the known range of neutrophil defense mechanisms and catalyzed a discipline of research focused upon ways in which neutrophils can shape the immunologic landscape of certain autoimmune diseases, including systemic lupus erythematosus. Enhanced neutrophil extracellular trap formation and impaired neutrophil extracellular trap clearance may contribute to immunogenicity in systemic lupus erythematosus and other autoimmune diseases by promoting the externalization of modified autoantigens, inducing synthesis of type I IFNs, stimulating the inflammasome, and activating both the classic and alternative pathways of the complement system. Vasculopathy is a central feature of many autoimmune diseases, and neutrophil extracellular traps may contribute directly to endothelial cell dysfunction, atherosclerotic plaque burden, and thrombosis. The elucidation of the subcellular events of neutrophil extracellular trap formation may generate novel, therapeutic strategies that target the innate immune system in autoimmune and vascular diseases.

  14. A Metabolic Shift toward Pentose Phosphate Pathway Is Necessary for Amyloid Fibril- and Phorbol 12-Myristate 13-Acetate-induced Neutrophil Extracellular Trap (NET) Formation.

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    Azevedo, Estefania P; Rochael, Natalia C; Guimarães-Costa, Anderson B; de Souza-Vieira, Thiago S; Ganilho, Juliana; Saraiva, Elvira M; Palhano, Fernando L; Foguel, Debora

    2015-09-01

    Neutrophils are the main defense cells of the innate immune system. Upon stimulation, neutrophils release their chromosomal DNA to trap and kill microorganisms and inhibit their dissemination. These chromatin traps are termed neutrophil extracellular traps (NETs) and are decorated with granular and cytoplasm proteins. NET release can be induced by several microorganism membrane components, phorbol 12-myristate 13-acetate as well as by amyloid fibrils, insoluble proteinaceous molecules associated with more than 40 different pathologies among other stimuli. The intracellular signaling involved in NET formation is complex and remains unclear for most tested stimuli. Herein we demonstrate that a metabolic shift toward the pentose phosphate pathway (PPP) is necessary for NET release because glucose-6-phosphate dehydrogenase (G6PD), an important enzyme from PPP, fuels NADPH oxidase with NADPH to produce superoxide and thus induce NETs. In addition, we observed that mitochondrial reactive oxygen species, which are NADPH-independent, are not effective in producing NETs. These data shed new light on how the PPP and glucose metabolism contributes to NET formation.

  15. Neutrophil Extracellular Traps and Microcrystals

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    2017-01-01

    Neutrophil extracellular traps represent a fascinating mechanism by which PMNs entrap extracellular microbes. The primary purpose of this innate immune mechanism is thought to localize the infection at an early stage. Interestingly, the ability of different microcrystals to induce NET formation has been recently described. Microcrystals are insoluble crystals with a size of 1–100 micrometers that have different composition and shape. Microcrystals have it in common that they irritate phagocytes including PMNs and typically trigger an inflammatory response. This review is the first to summarize observations with regard to PMN activation and NET release induced by microcrystals. Gout-causing monosodium urate crystals, pseudogout-causing calcium pyrophosphate dehydrate crystals, cholesterol crystals associated with atherosclerosis, silicosis-causing silica crystals, and adjuvant alum crystals are discussed. PMID:28373994

  16. Neutrophil Extracellular Traps and Microcrystals.

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    Rada, Balázs

    2017-01-01

    Neutrophil extracellular traps represent a fascinating mechanism by which PMNs entrap extracellular microbes. The primary purpose of this innate immune mechanism is thought to localize the infection at an early stage. Interestingly, the ability of different microcrystals to induce NET formation has been recently described. Microcrystals are insoluble crystals with a size of 1-100 micrometers that have different composition and shape. Microcrystals have it in common that they irritate phagocytes including PMNs and typically trigger an inflammatory response. This review is the first to summarize observations with regard to PMN activation and NET release induced by microcrystals. Gout-causing monosodium urate crystals, pseudogout-causing calcium pyrophosphate dehydrate crystals, cholesterol crystals associated with atherosclerosis, silicosis-causing silica crystals, and adjuvant alum crystals are discussed.

  17. Neutrophil Extracellular Traps and Microcrystals

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    Balázs Rada

    2017-01-01

    Full Text Available Neutrophil extracellular traps represent a fascinating mechanism by which PMNs entrap extracellular microbes. The primary purpose of this innate immune mechanism is thought to localize the infection at an early stage. Interestingly, the ability of different microcrystals to induce NET formation has been recently described. Microcrystals are insoluble crystals with a size of 1–100 micrometers that have different composition and shape. Microcrystals have it in common that they irritate phagocytes including PMNs and typically trigger an inflammatory response. This review is the first to summarize observations with regard to PMN activation and NET release induced by microcrystals. Gout-causing monosodium urate crystals, pseudogout-causing calcium pyrophosphate dehydrate crystals, cholesterol crystals associated with atherosclerosis, silicosis-causing silica crystals, and adjuvant alum crystals are discussed.

  18. A novel method for high-throughput detection and quantification of neutrophil extracellular traps reveals ROS-independent NET release with immune complexes.

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    Kraaij, Tineke; Tengström, Fredrik C; Kamerling, Sylvia W A; Pusey, Charles D; Scherer, H Ulrich; Toes, Rene E M; Rabelink, Ton J; van Kooten, Cees; Teng, Y K Onno

    2016-06-01

    A newly-described first-line immune defence mechanism of neutrophils is the release of neutrophil extracellular traps (NETs). Immune complexes (ICxs) induce low level NET release. As such, the in vitro quantification of NETs is challenging with current methodologies. In order to investigate the role of NET release in ICx-mediated autoimmune diseases, we developed a highly sensitive and automated method for quantification of NETs. After labelling human neutrophils with PKH26 and extracellular DNA with Sytox green, cells are fixed and automatically imaged with 3-dimensional confocal laser scanning microscopy (3D-CLSM). NET release is then quantified with digital image analysis whereby the NET amount (Sytox green area) is corrected for the number of imaged neutrophils (PKH26 area). A high sensitivity of the assay is achieved by a) significantly augmenting the area of the well imaged (11%) as compared to conventional assays (0.5%) and b) using a 3D imaging technique for optimal capture of NETs, which are topologically superimposed on neutrophils. In this assay, we confirmed low levels of NET release upon human ICx stimulation which were positive for citrullinated histones and neutrophil elastase. In contrast to PMA-induced NET release, ICx-induced NET release was unchanged when co-incubated with diphenyleneiodonium (DPI). We were able to quantify NET release upon stimulation with serum from RA and SLE patients, which was not observed with normal human serum. To our knowledge, this is the first semi-automated assay capable of sensitive detection and quantification of NET release at a low threshold by using 3D CLSM. The assay is applicable in a high-throughput manner and allows the in vitro analysis of NET release in ICx-mediated autoimmune diseases.

  19. Neutrophil extracellular trap (NET)-mediated killing of Pseudomonas aeruginosa: evidence of acquired resistance within the CF airway, independent of CFTR.

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    Young, Robert L; Malcolm, Kenneth C; Kret, Jennifer E; Caceres, Silvia M; Poch, Katie R; Nichols, David P; Taylor-Cousar, Jennifer L; Saavedra, Milene T; Randell, Scott H; Vasil, Michael L; Burns, Jane L; Moskowitz, Samuel M; Nick, Jerry A

    2011-01-01

    The inability of neutrophils to eradicate Pseudomonas aeruginosa within the cystic fibrosis (CF) airway eventually results in chronic infection by the bacteria in nearly 80 percent of patients. Phagocytic killing of P. aeruginosa by CF neutrophils is impaired due to decreased cystic fibrosis transmembrane conductance regulator (CFTR) function and virulence factors acquired by the bacteria. Recently, neutrophil extracellular traps (NETs), extracellular structures composed of neutrophil chromatin complexed with granule contents, were identified as an alternative mechanism of pathogen killing. The hypothesis that NET-mediated killing of P. aeruginosa is impaired in the context of the CF airway was tested. P. aeruginosa induced NET formation by neutrophils from healthy donors in a bacterial density dependent fashion. When maintained in suspension through continuous rotation, P. aeruginosa became physically associated with NETs. Under these conditions, NETs were the predominant mechanism of killing, across a wide range of bacterial densities. Peripheral blood neutrophils isolated from CF patients demonstrated no impairment in NET formation or function against P. aeruginosa. However, isogenic clinical isolates of P. aeruginosa obtained from CF patients early and later in the course of infection demonstrated an acquired capacity to withstand NET-mediated killing in 8 of 9 isolates tested. This resistance correlated with development of the mucoid phenotype, but was not a direct result of the excess alginate production that is characteristic of mucoidy. Together, these results demonstrate that neutrophils can kill P. aeruginosa via NETs, and in vitro this response is most effective under non-stationary conditions with a low ratio of bacteria to neutrophils. NET-mediated killing is independent of CFTR function or bacterial opsonization. Failure of this response in the context of the CF airway may occur, in part, due to an acquired resistance against NET-mediated killing by

  20. Neutrophil extracellular trap (NET-mediated killing of Pseudomonas aeruginosa: evidence of acquired resistance within the CF airway, independent of CFTR.

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    Robert L Young

    Full Text Available The inability of neutrophils to eradicate Pseudomonas aeruginosa within the cystic fibrosis (CF airway eventually results in chronic infection by the bacteria in nearly 80 percent of patients. Phagocytic killing of P. aeruginosa by CF neutrophils is impaired due to decreased cystic fibrosis transmembrane conductance regulator (CFTR function and virulence factors acquired by the bacteria. Recently, neutrophil extracellular traps (NETs, extracellular structures composed of neutrophil chromatin complexed with granule contents, were identified as an alternative mechanism of pathogen killing. The hypothesis that NET-mediated killing of P. aeruginosa is impaired in the context of the CF airway was tested. P. aeruginosa induced NET formation by neutrophils from healthy donors in a bacterial density dependent fashion. When maintained in suspension through continuous rotation, P. aeruginosa became physically associated with NETs. Under these conditions, NETs were the predominant mechanism of killing, across a wide range of bacterial densities. Peripheral blood neutrophils isolated from CF patients demonstrated no impairment in NET formation or function against P. aeruginosa. However, isogenic clinical isolates of P. aeruginosa obtained from CF patients early and later in the course of infection demonstrated an acquired capacity to withstand NET-mediated killing in 8 of 9 isolates tested. This resistance correlated with development of the mucoid phenotype, but was not a direct result of the excess alginate production that is characteristic of mucoidy. Together, these results demonstrate that neutrophils can kill P. aeruginosa via NETs, and in vitro this response is most effective under non-stationary conditions with a low ratio of bacteria to neutrophils. NET-mediated killing is independent of CFTR function or bacterial opsonization. Failure of this response in the context of the CF airway may occur, in part, due to an acquired resistance against NET

  1. Rab27a is essential for the formation of neutrophil extracellular traps (NETs in neutrophil-like differentiated HL60 cells.

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    Tatsumi Kawakami

    Full Text Available Neutrophils play a crucial role in host defence. In response to a variety of inflammatory stimulation, they form neutrophil extracellular traps (NETs. NETs are extracellular structures composed of chromatin fibers decorated with antimicrobial proteins and developing studies indicate that NETs contribute to extracellular microbial killing. While the intracellular signaling pathways that regulate NET formation remain largely unknown, there is growing evidence that generation of reactive oxygen species (ROS is a key event for NET formation. The Rab family small GTPase Rab27a is an important component of the secretory machinery of azurophilic granules in neutrophils. However, the precise mechanism of NET formation and whether or not Rab27a contributes to this process are unknown. Using neutrophil-like differentiated HL60 cells, we show here that Rab27a plays an essential role in both phorbol myristate acetate (PMA- and Candida albicans-induced NET formation by regulating ROS production. Rab27a-knockdown inhibited ROS-positive phagosome formation during complement-mediated phagocytosis. To investigate the role of Rab27a in neutrophil function in detail, both primary human neutrophils and neutrophil-like differentiated HL60 cells were treated with PMA, and NET formation process was assessed by measurement of release of histone H3 into the medium, citrullination of the arginine in position 3 of histone H4 and chase of the nuclear change of the living cells in the co-existence of both cell-permeable and -impermeable nuclear indicators. PMA-induced NET formation occured sequentially in both neutrophil-like differentiated HL60 cells and primary neutrophils, and Rab27a-knockdown clearly inhibited NET formation in association with reduced ROS production. We also found that serum-treated Candida albicans triggers NET formation in a ROS-dependent manner, and that Rab27a-knockdown inhibits this process as well. Our findings demonstrate that Rab27a plays an

  2. Neutrophil extracellular traps in tissue pathology.

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    Nakazawa, Daigo; Kumar, Santosh; Desai, Jyaysi; Anders, Hans-Joachim

    2017-03-01

    Neutrophil extracellular traps (NETs) are innate immune systems against invading pathogens. NETs are characterized as released DNA mixed with cytoplasmic antimicrobial proteins such as myeloperoxidase, proteinase3 and neutrophil elastase. While NETs are thought to have an important role in host defense, recent work has suggested that NETs contribute to tissue injury in non-infectious disease states. Uncontrolled NET formation in autoimmune diseases, metabolic disorders, cancers and thrombotic diseases can exacerbate a disease or even be a major initiator of tissue injury. But spotting NETs in tissues is not easy. Here we review the available histopathological evidence on the presence of NETs in a variety of diseases. We discuss technical difficulties and potential sources of misinterpretation while trying to detect NETs in tissue samples.

  3. Neutrophils cast extracellular traps in response to protozoan parasites.

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    Abi Abdallah, Delbert S; Denkers, Eric Y

    2012-01-01

    Release of extracellular traps by neutrophils is a now well-established phenomenon that contributes to the innate response to extracellular bacterial and fungal pathogens. The importance of NETs during protozoan infection has been less explored, but recent findings suggest an emerging role for release of neutrophil-derived extracellular DNA in response to this class of microbial pathogens. The present review summarizes findings to date regarding elicitation of NETs by Toxoplasma gondii, Plasmodium falciparum, Eimeria bovis, and Leishmania spp.

  4. Signal Inhibitory Receptor on Leukocytes-1 Limits the Formation of Neutrophil Extracellular Traps, but Preserves Intracellular Bacterial Killing

    NARCIS (Netherlands)

    Van Avondt, Kristof; van der Linden, Maarten; Naccache, Paul H; Egan, David A; Meyaard, Linde

    2016-01-01

    In response to microbial invasion, neutrophils release neutrophil extracellular traps (NETs) to trap and kill extracellular microbes. Alternatively, NET formation can result in tissue damage in inflammatory conditions and may perpetuate autoimmune disease. Intervention strategies that are aimed at m

  5. Metabolic requirements for neutrophil extracellular traps formation

    Science.gov (United States)

    Rodríguez-Espinosa, Oscar; Rojas-Espinosa, Oscar; Moreno-Altamirano, María Maximina Bertha; López-Villegas, Edgar Oliver; Sánchez-García, Francisco Javier

    2015-01-01

    As part of the innate immune response, neutrophils are at the forefront of defence against infection, resolution of inflammation and wound healing. They are the most abundant leucocytes in the peripheral blood, have a short lifespan and an estimated turnover of 1010 to 1011 cells per day. Neutrophils efficiently clear microbial infections by phagocytosis and by oxygen-dependent and oxygen-independent mechanisms. In 2004, a new neutrophil anti-microbial mechanism was described, the release of neutrophil extracellular traps (NETs) composed of DNA, histones and anti-microbial peptides. Several microorganisms, bacterial products, as well as pharmacological stimuli such as PMA, were shown to induce NETs. Neutrophils contain relatively few mitochondria, and derive most of their energy from glycolysis. In this scenario we aimed to analyse some of the metabolic requirements for NET formation. Here it is shown that NETs formation is strictly dependent on glucose and to a lesser extent on glutamine, that Glut-1, glucose uptake, and glycolysis rate increase upon PMA stimulation, and that NET formation is inhibited by the glycolysis inhibitor, 2-deoxy-glucose, and to a lesser extent by the ATP synthase inhibitor oligomycin. Moreover, when neutrophils were exposed to PMA in glucose-free medium for 3 hr, they lost their characteristic polymorphic nuclei but did not release NETs. However, if glucose (but not pyruvate) was added at this time, NET release took place within minutes, suggesting that NET formation could be metabolically divided into two phases; the first, independent from exogenous glucose (chromatin decondensation) and, the second (NET release), strictly dependent on exogenous glucose and glycolysis. PMID:25545227

  6. Neutrophil extracellular traps - the dark side of neutrophils

    DEFF Research Database (Denmark)

    Sørensen, Ole E.; Borregaard, Niels

    2016-01-01

    Neutrophil extracellular traps (NETs) were discovered as extracellular strands of decondensed DNA in complex with histones and granule proteins, which were expelled from dying neutrophils to ensnare and kill microbes. NETs are formed during infection in vivo by mechanisms different from those...... originally described in vitro. Citrullination of histones by peptidyl arginine deiminase 4 (PAD4) is central for NET formation in vivo. NETs may spur formation of autoantibodies and may also serve as scaffolds for thrombosis, thereby providing a link among infection, autoimmunity, and thrombosis...

  7. Killing by neutrophil extracellular traps: fact or folklore?

    Science.gov (United States)

    Menegazzi, Renzo; Decleva, Eva; Dri, Pietro

    2012-02-02

    Neutrophil extracellular traps (NETs) are DNA structures released by dying neutrophils and claimed to constitute a new microbicidal mechanism. Killing by NET-forming cells is ascribed to these structures because it is prevented by preincubation with DNase, which has been shown to dismantle NETs, before addition of the target microorganisms. Curiously, the possibility that the microorganisms ensnared in NETs are alive has not been considered. Using Staphylococcus aureus and Candida albicans blastospores, we demonstrate that the microorganisms captured by NETs and thought to be killed are alive because they are released and recovered in cell medium by incubation with DNase. It is concluded that NETs entrap but do not kill microbes.

  8. Lactoferrin Suppresses Neutrophil Extracellular Traps Release in Inflammation.

    Science.gov (United States)

    Okubo, Koshu; Kamiya, Mako; Urano, Yasuteru; Nishi, Hiroshi; Herter, Jan M; Mayadas, Tanya; Hirohama, Daigoro; Suzuki, Kazuo; Kawakami, Hiroshi; Tanaka, Mototsugu; Kurosawa, Miho; Kagaya, Shinji; Hishikawa, Keiichi; Nangaku, Masaomi; Fujita, Toshiro; Hayashi, Matsuhiko; Hirahashi, Junichi

    2016-08-01

    Neutrophils are central players in the innate immune system. They generate neutrophil extracellular traps (NETs), which protect against invading pathogens but are also associated with the development of autoimmune and/or inflammatory diseases and thrombosis. Here, we report that lactoferrin, one of the components of NETs, translocated from the cytoplasm to the plasma membrane and markedly suppressed NETs release. Furthermore, exogenous lactoferrin shrunk the chromatin fibers found in released NETs, without affecting the generation of oxygen radicals, but this failed after chemical removal of the positive charge of lactoferrin, suggesting that charge-charge interactions between lactoferrin and NETs were required for this function. In a model of immune complex-induced NET formation in vivo, intravenous lactoferrin injection markedly reduced the extent of NET formation. These observations suggest that lactoferrin serves as an intrinsic inhibitor of NETs release into the circulation. Thus, lactoferrin may represent a therapeutic lead for controlling NETs release in autoimmune and/or inflammatory diseases.

  9. Neutrophil Extracellular Traps in Periodontitis: A Web of Intrigue.

    Science.gov (United States)

    White, P C; Chicca, I J; Cooper, P R; Milward, M R; Chapple, I L C

    2016-01-01

    Neutrophil extracellular traps (NETs) represent a novel paradigm in neutrophil-mediated immunity. NETs are believed to constitute a highly conserved antimicrobial strategy comprising decondensed nuclear DNA and associated histones that are extruded into the extracellular space. Associated with the web-like strands of DNA is an array of antimicrobial peptides (AMPs), which facilitate the extracellular destruction of microorganisms that become entrapped within the NETs. NETs can be released by cells that remain viable or following a unique form of programmed cell death known as NETosis, which is dependent on the production of reactive oxygen species (ROS) and the decondensing of the nuclear DNA catalyzed by peptidyl arginine deiminase-4. NETs are produced in response to a range of pathogens, including bacteria, viruses, fungi, and protozoa, as well as host-derived mediators. NET release is, however, not without cost, as the concomitant release of cytotoxic molecules can also cause host tissue damage. This is evidenced by a number of immune-mediated diseases, in which excess or dysfunctional NET production, bacterial NET evasion, and decreased NET removal are associated with disease pathogenesis. Periodontitis is the most prevalent infectious-inflammatory disease of humans, characterized by a dysregulated neutrophilic response to specific bacterial species within the subgingival plaque biofilm. Neutrophils are the predominant inflammatory cell involved in periodontitis and have previously been found to exhibit hyperactivity and hyperreactivity in terms of ROS production in chronic periodontitis patients. However, the contribution of ROS-dependent NET formation to periodontal health or disease remains unclear. In this focused review, we discuss the mechanisms, stimuli, and requirements for NET production; the ability of NET-DNA and NET-associated AMPs to entrap and kill pathogens; and the potential immunogenicity of NETs in disease. We also speculate on the potential

  10. Neutrophil extracellular traps involvement in corneal fungal infection

    Science.gov (United States)

    Zhao, Yingying; Zhang, Fan; Wan, Ting; Fan, Fangli; Xie, Xin; Lin, Zhenyun

    2016-01-01

    Purpose Neutrophils release neutrophil extracellular traps (NETs) when defending against invading microorganisms. We investigated the existence of NETs in fungal keratitis. Methods Fourteen patients with unilateral fungal keratitis were included. Detailed information about each patient was recorded, including (1) patient history (onset of symptoms and previous therapy), (2) ocular examination findings by slit-lamp biomicroscopy, (3) laboratory findings from direct smear examination and culture of corneal scrapings, (4) NET formation, and (5) treatment strategy and prognosis. Immunofluorescence staining was used to evaluate the existence of NETs on corneal scrapings. The relationship between the quantification of NETs and the clinical character of the fungal keratitis was identified. Results NETs were identified in all 14 patients. Patients with a higher grade of NET formation and fewer fungal hyphae always showed a good treatment response and a short course of infection. NETs were consistently found mixed with fungal hyphae in the corneal scrapings from infected patients. No statistical significance was found between the grade of NETs formed and the course of infection before presentation, and no relationship between the quantification of NETs and the size of the ulcer was found. Conclusions The results suggest that NETs are involved in fungal keratitis. The number of NETs in infected corneas may provide a tool for evaluating the prognosis for fungal keratitis. PMID:27559290

  11. Vibrio cholerae evades neutrophil extracellular traps by the activity of two extracellular nucleases.

    Directory of Open Access Journals (Sweden)

    Andrea Seper

    Full Text Available The Gram negative bacterium Vibrio cholerae is the causative agent of the secretory diarrheal disease cholera, which has traditionally been classified as a noninflammatory disease. However, several recent reports suggest that a V. cholerae infection induces an inflammatory response in the gastrointestinal tract indicated by recruitment of innate immune cells and increase of inflammatory cytokines. In this study, we describe a colonization defect of a double extracellular nuclease V. cholerae mutant in immunocompetent mice, which is not evident in neutropenic mice. Intrigued by this observation, we investigated the impact of neutrophils, as a central part of the innate immune system, on the pathogen V. cholerae in more detail. Our results demonstrate that V. cholerae induces formation of neutrophil extracellular traps (NETs upon contact with neutrophils, while V. cholerae in return induces the two extracellular nucleases upon presence of NETs. We show that the V. cholerae wild type rapidly degrades the DNA component of the NETs by the combined activity of the two extracellular nucleases Dns and Xds. In contrast, NETs exhibit prolonged stability in presence of the double nuclease mutant. Finally, we demonstrate that Dns and Xds mediate evasion of V. cholerae from NETs and lower the susceptibility for extracellular killing in the presence of NETs. This report provides a first comprehensive characterization of the interplay between neutrophils and V. cholerae along with new evidence that the innate immune response impacts the colonization of V. cholerae in vivo. A limitation of this study is an inability for technical and physiological reasons to visualize intact NETs in the intestinal lumen of infected mice, but we can hypothesize that extracellular nuclease production by V. cholerae may enhance survival fitness of the pathogen through NET degradation.

  12. 36 CFR 2.4 - Weapons, traps and nets.

    Science.gov (United States)

    2010-07-01

    ... 36 Parks, Forests, and Public Property 1 2010-07-01 2010-07-01 false Weapons, traps and nets. 2.4... PROTECTION, PUBLIC USE AND RECREATION § 2.4 Weapons, traps and nets. (a)(1) Except as otherwise provided in... prohibited: (i) Possessing a weapon, trap or net (ii) Carrying a weapon, trap or net (iii) Using a weapon...

  13. Carp neutrophilic granulocyes form extracellular traps via ROS-dependent and independent pathways

    NARCIS (Netherlands)

    Pijanowski, L.; Golbach, L.A.; Kolaczkowska, E.; Scheer, M.H.; Verburg-van Kemenade, B.M.L.; Chadzinska, M.K.

    2013-01-01

    Neutrophil extracellular traps (NETs) have recently been described as an important innate defense mechanism that leads to immobilization and killing of invading pathogens. NETs have been identified in several species, but the mechanisms involved in NET formation and their role in infection have not

  14. DNA is an antimicrobial component of neutrophil extracellular traps.

    Directory of Open Access Journals (Sweden)

    Tyler W R Halverson

    2015-01-01

    Full Text Available Neutrophil extracellular traps (NETs comprise an ejected lattice of chromatin enmeshed with granular and nuclear proteins that are capable of capturing and killing microbial invaders. Although widely employed to combat infection, the antimicrobial mechanism of NETs remains enigmatic. Efforts to elucidate the bactericidal component of NETs have focused on the role of NET-bound proteins including histones, calprotectin and cathepsin G protease; however, exogenous and microbial derived deoxyribonuclease (DNase remains the most potent inhibitor of NET function. DNA possesses a rapid bactericidal activity due to its ability to sequester surface bound cations, disrupt membrane integrity and lyse bacterial cells. Here we demonstrate that direct contact and the phosphodiester backbone are required for the cation chelating, antimicrobial property of DNA. By treating NETs with excess cations or phosphatase enzyme, the antimicrobial activity of NETs is neutralized, but NET structure, including the localization and function of NET-bound proteins, is maintained. Using intravital microscopy, we visualized NET-like structures in the skin of a mouse during infection with Pseudomonas aeruginosa. Relative to other bacteria, P. aeruginosa is a weak inducer of NETosis and is more resistant to NETs. During NET exposure, we demonstrate that P. aeruginosa responds by inducing the expression of surface modifications to defend against DNA-induced membrane destabilization and NET-mediated killing. Further, we show induction of this bacterial response to NETs is largely due to the bacterial detection of DNA. Therefore, we conclude that the DNA backbone contributes both to the antibacterial nature of NETs and as a signal perceived by microbes to elicit host-resistance strategies.

  15. Entamoeba histolytica Trophozoites and Lipopeptidophosphoglycan Trigger Human Neutrophil Extracellular Traps.

    Science.gov (United States)

    Ávila, Eva E; Salaiza, Norma; Pulido, Julieta; Rodríguez, Mayra C; Díaz-Godínez, César; Laclette, Juan P; Becker, Ingeborg; Carrero, Julio C

    2016-01-01

    Neutrophil defense mechanisms include phagocytosis, degranulation and the formation of extracellular traps (NET). These networks of DNA are triggered by several immune and microbial factors, representing a defense strategy to prevent microbial spread by trapping/killing pathogens. This may be important against Entamoeba histolytica, since its large size hinders its phagocytosis. The aim of this study was to determine whether E. histolytica and their lipopeptidophosphoglycan (EhLPPG) induce the formation of NETs and the outcome of their interaction with the parasite. Our data show that live amoebae and EhLPPG, but not fixed trophozoites, induced NET formation in a time and dose dependent manner, starting at 5 min of co-incubation. Although immunofluorescence studies showed that the NETs contain cathelicidin LL-37 in close proximity to amoebae, the trophozoite growth was only affected when ethylene glycol tetra-acetic acid (EGTA) was present during contact with NETs, suggesting that the activity of enzymes requiring calcium, such as DNases, may be important for amoeba survival. In conclusion, E. histolytica trophozoites and EhLPPG induce in vitro formation of human NETs, which did not affect the parasite growth unless a chelating agent was present. These results suggest that NETs may be an important factor of the innate immune response during infection with E. histolytica.

  16. Entamoeba histolytica Trophozoites and Lipopeptidophosphoglycan Trigger Human Neutrophil Extracellular Traps

    Science.gov (United States)

    Ávila, Eva E.; Rodríguez, Mayra C.; Díaz-Godínez, César; Laclette, Juan P.; Becker, Ingeborg; Carrero, Julio C.

    2016-01-01

    Neutrophil defense mechanisms include phagocytosis, degranulation and the formation of extracellular traps (NET). These networks of DNA are triggered by several immune and microbial factors, representing a defense strategy to prevent microbial spread by trapping/killing pathogens. This may be important against Entamoeba histolytica, since its large size hinders its phagocytosis. The aim of this study was to determine whether E. histolytica and their lipopeptidophosphoglycan (EhLPPG) induce the formation of NETs and the outcome of their interaction with the parasite. Our data show that live amoebae and EhLPPG, but not fixed trophozoites, induced NET formation in a time and dose dependent manner, starting at 5 min of co-incubation. Although immunofluorescence studies showed that the NETs contain cathelicidin LL-37 in close proximity to amoebae, the trophozoite growth was only affected when ethylene glycol tetra-acetic acid (EGTA) was present during contact with NETs, suggesting that the activity of enzymes requiring calcium, such as DNases, may be important for amoeba survival. In conclusion, E. histolytica trophozoites and EhLPPG induce in vitro formation of human NETs, which did not affect the parasite growth unless a chelating agent was present. These results suggest that NETs may be an important factor of the innate immune response during infection with E. histolytica. PMID:27415627

  17. Lactoferrin Suppresses Neutrophil Extracellular Traps Release in Inflammation

    Directory of Open Access Journals (Sweden)

    Koshu Okubo

    2016-08-01

    Full Text Available Neutrophils are central players in the innate immune system. They generate neutrophil extracellular traps (NETs, which protect against invading pathogens but are also associated with the development of autoimmune and/or inflammatory diseases and thrombosis. Here, we report that lactoferrin, one of the components of NETs, translocated from the cytoplasm to the plasma membrane and markedly suppressed NETs release. Furthermore, exogenous lactoferrin shrunk the chromatin fibers found in released NETs, without affecting the generation of oxygen radicals, but this failed after chemical removal of the positive charge of lactoferrin, suggesting that charge-charge interactions between lactoferrin and NETs were required for this function. In a model of immune complex-induced NET formation in vivo, intravenous lactoferrin injection markedly reduced the extent of NET formation. These observations suggest that lactoferrin serves as an intrinsic inhibitor of NETs release into the circulation. Thus, lactoferrin may represent a therapeutic lead for controlling NETs release in autoimmune and/or inflammatory diseases.

  18. 36 CFR 1002.4 - Weapons, traps and nets.

    Science.gov (United States)

    2010-07-01

    ... 36 Parks, Forests, and Public Property 3 2010-07-01 2010-07-01 false Weapons, traps and nets. 1002... AND RECREATION § 1002.4 Weapons, traps and nets. (a)(1) Except as otherwise provided in this section, the following are prohibited: (i) Possessing a weapon, trap or net. (ii) Carrying a weapon, trap or...

  19. Human neutrophils produce extracellular traps against Paracoccidioides brasiliensis.

    Science.gov (United States)

    Mejía, Susana P; Cano, Luz E; López, Juan A; Hernandez, Orville; González, Ángel

    2015-05-01

    Neutrophils play an important role as effector cells and contribute to the resistance of the host against microbial pathogens. Neutrophils are able to produce extracellular traps (NETs) in response to medically important fungi, including Aspergillus spp., Candida albicans and Cryptococcus gattii. However, NET production in response to Paracoccidioides brasiliensis has yet to be studied. We have demonstrated that human neutrophils produce NETs against both conidia and yeasts of P. brasiliensis. Although the NADPH oxidase inhibitor diphenyleneiodonium chloride (DPI) did not alter NET production against conidia, it partially suppressed NET formation against P. brasiliensis yeasts. Cytochalasin D or IFN-γ did not affect the production of NETs against the fungus. Additionally, a mutant strain of P. brasiliensis with reduced expression of an alternative oxidase induced significantly higher levels of NETs in comparison with the WT strain. Finally, c.f.u. quantification of P. brasiliensis showed no significant differences when neutrophils were treated with DPI, DNase I or cytochalasin D as compared with untreated cells. These data establish that NET formation by human neutrophils appears to be either dependent or independent of reactive oxygen species production, correlating with the fungal morphotype used for stimulation. However, this mechanism was ineffective in killing the fungus.

  20. P-selectin promotes neutrophil extracellular trap formation in mice.

    Science.gov (United States)

    Etulain, Julia; Martinod, Kimberly; Wong, Siu Ling; Cifuni, Stephen M; Schattner, Mirta; Wagner, Denisa D

    2015-07-01

    Neutrophil extracellular traps (NETs) can be released in the vasculature. In addition to trapping microbes, they promote inflammatory and thrombotic diseases. Considering that P-selectin induces prothrombotic and proinflammatory signaling, we studied the role of this selectin in NET formation. NET formation (NETosis) was induced by thrombin-activated platelets rosetting with neutrophils and was inhibited by anti-P-selectin aptamer or anti-P-selectin glycoprotein ligand-1 (PSGL-1) inhibitory antibody but was not induced by platelets from P-selectin(-/-) mice. Moreover, NETosis was also promoted by P-selectin-immunoglobulin fusion protein but not by control immunoglobulin. We isolated neutrophils from mice engineered to overproduce soluble P-selectin (P-selectin(ΔCT/ΔCT) mice). Although the levels of circulating DNA and nucleosomes (indicative of spontaneous NETosis) were normal in these mice, basal neutrophil histone citrullination and presence of P-selectin on circulating neutrophils were elevated. NET formation after stimulation with platelet activating factor, ionomycin, or phorbol 12-myristate 13-acetate was significantly enhanced, indicating that the P-selectin(ΔCT/ΔCT) neutrophils were primed for NETosis. In summary, P-selectin, cellular or soluble, through binding to PSGL-1, promotes NETosis, suggesting that this pathway is a potential therapeutic target for NET-related diseases.

  1. Influence of complement on neutrophil extracellular trap release induced by bacteria

    DEFF Research Database (Denmark)

    Palmer, Lisa Joanne; Damgaard, Christian; Holmstrup, Palle;

    2016-01-01

    Background and Objectives Neutrophil extracellular trap (NET) release has generally been studied in the absence of serum, or at low concentrations of untreated or heat-inactivated serum. The influence of serum complement on NET release therefore remains unclear. We examined the DNA release induce...

  2. 36 CFR 13.30 - Weapons, traps and nets.

    Science.gov (United States)

    2010-07-01

    ... 36 Parks, Forests, and Public Property 1 2010-07-01 2010-07-01 false Weapons, traps and nets. 13... INTERIOR NATIONAL PARK SYSTEM UNITS IN ALASKA General Provisions § 13.30 Weapons, traps and nets. (a... of this chapter, the following are prohibited— (1) Possessing a weapon, trap, or net; (2) Carrying a...

  3. Autophagy Primes Neutrophils for Neutrophil Extracellular Trap Formation during Sepsis.

    Science.gov (United States)

    Park, So Young; Shrestha, Sanjeeb; Youn, Young-Jin; Kim, Jun-Kyu; Kim, Shin-Yeong; Kim, Hyun Jung; Park, So-Hee; Ahn, Won-Gyun; Kim, Shin; Lee, Myung Goo; Jung, Ki-Suck; Park, Yong Bum; Mo, Eun-Kyung; Ko, Yousang; Lee, Suh-Young; Koh, Younsuck; Park, Myung Jae; Song, Dong-Keun; Hong, Chang-Won

    2017-09-01

    Neutrophils are key effectors in the host's immune response to sepsis. Excessive stimulation or dysregulated neutrophil functions are believed to be responsible for sepsis pathogenesis. However, the mechanisms regulating functional plasticity of neutrophils during sepsis have not been fully determined. We investigated the role of autophagy in neutrophil functions during sepsis in patients with community-acquired pneumonia. Neutrophils were isolated from patients with sepsis and stimulated with phorbol 12-myristate 13-acetate (PMA). The levels of reactive oxygen species generation, neutrophil extracellular trap (NET) formation, and granule release, and the autophagic status were evaluated. The effect of neutrophil autophagy augmentation was further evaluated in a mouse model of sepsis. Neutrophils isolated from patients who survived sepsis showed an increase in autophagy induction, and were primed for NET formation in response to subsequent PMA stimulation. In contrast, neutrophils isolated from patients who did not survive sepsis showed dysregulated autophagy and a decreased response to PMA stimulation. The induction of autophagy primed healthy neutrophils for NET formation and vice versa. In a mouse model of sepsis, the augmentation of autophagy improved survival via a NET-dependent mechanism. These results indicate that neutrophil autophagy primes neutrophils for increased NET formation, which is important for proper neutrophil effector functions during sepsis. Our study provides important insights into the role of autophagy in neutrophils during sepsis.

  4. A thermonuclease of Neisseria gonorrhoeae enhances bacterial escape from killing by neutrophil extracellular traps.

    Science.gov (United States)

    Juneau, Richard A; Stevens, Jacqueline S; Apicella, Michael A; Criss, Alison K

    2015-07-15

    Acute gonorrhea is characterized by neutrophilic inflammation that is insufficient to clear Neisseria gonorrhoeae. Activated neutrophils release extracellular traps (NETs), which are composed of chromatin and decorated with antimicrobial proteins. The N. gonorrhoeae NG0969 open reading frame contains a gene (nuc) that encodes a putatively secreted thermonuclease (Nuc) that contributes to biofilm remodeling. Here, we report that Nuc degrades NETs to help N. gonorrhoeae resist killing by neutrophils. Primary human neutrophils released NETs after exposure to N. gonorrhoeae, but NET integrity declined over time with Nuc-containing bacteria. Recombinant Nuc and conditioned medium from Nuc-containing N. gonorrhoeae degraded human neutrophil DNA and NETs. NETs were found to have antimicrobial activity against N. gonorrhoeae, and Nuc expression enhanced N. gonorrhoeae survival in the presence of neutrophils that released NETs. We propose that Nuc enables N. gonorrhoeae to escape trapping and killing by NETs during symptomatic infection, highlighting Nuc as a multifunctional virulence factor for N. gonorrhoeae.

  5. High Throughput Measurement of Extracellular DNA Release and Quantitative NET Formation in Human Neutrophils In Vitro.

    Science.gov (United States)

    Sil, Payel; Yoo, Dae-Goon; Floyd, Madison; Gingerich, Aaron; Rada, Balazs

    2016-06-18

    Neutrophil granulocytes are the most abundant leukocytes in the human blood. Neutrophils are the first to arrive at the site of infection. Neutrophils developed several antimicrobial mechanisms including phagocytosis, degranulation and formation of neutrophil extracellular traps (NETs). NETs consist of a DNA scaffold decorated with histones and several granule markers including myeloperoxidase (MPO) and human neutrophil elastase (HNE). NET release is an active process involving characteristic morphological changes of neutrophils leading to expulsion of their DNA into the extracellular space. NETs are essential to fight microbes, but uncontrolled release of NETs has been associated with several disorders. To learn more about the clinical relevance and the mechanism of NET formation, there is a need to have reliable tools capable of NET quantitation. Here three methods are presented that can assess NET release from human neutrophils in vitro. The first one is a high throughput assay to measure extracellular DNA release from human neutrophils using a membrane impermeable DNA-binding dye. In addition, two other methods are described capable of quantitating NET formation by measuring levels of NET-specific MPO-DNA and HNE-DNA complexes. These microplate-based methods in combination provide great tools to efficiently study the mechanism and regulation of NET formation of human neutrophils.

  6. Synchronized integrin engagement and chemokine activation is crucial in neutrophil extracellular trap-mediated sterile inflammation.

    Science.gov (United States)

    Rossaint, Jan; Herter, Jan M; Van Aken, Hugo; Napirei, Markus; Döring, Yvonne; Weber, Christian; Soehnlein, Oliver; Zarbock, Alexander

    2014-04-17

    There is emerging evidence that neutrophil extracellular traps (NETs) play important roles in inflammatory processes. Here we report that neutrophils have to be simultaneously activated by integrin-mediated outside-in- and G-protein-coupled receptor (GPCR) signaling to induce NET formation in acute lung injury (ALI), which is associated with a high mortality rate in critically ill patients. NETs consist of decondensed chromatin decorated with granular and cytosolic proteins and they can trap extracellular pathogens. The prerequisite for NET formation is the activation of neutrophils and the release of their DNA. In a neutrophil- and platelet-dependent mouse model of ventilator-induced lung injury (VILI), NETs were found in the lung microvasculature, and circulating NET components increased in the plasma. In this model, blocking integrin-mediated outside-in or either GPCR-signaling or heteromerization of platelet chemokines decreased NET formation and lung injury. Targeting NET components by DNAse1 application or neutrophil elastase-deficient mice protected mice from ALI, whereas DNase1(-/-)/Trap1(m/m) mice had an aggravated ALI, suggesting that NETs directly influence the severity of ALI. These data suggest that NETs form in the lungs during VILI, contribute to the disease process, and thus may be a promising new direction for the treatment of ALI.

  7. In vivo characterization of neutrophil extracellular traps in various organs of a murine sepsis model.

    Directory of Open Access Journals (Sweden)

    Koji Tanaka

    Full Text Available Neutrophil extracellular traps (NETs represent extracellular microbial trapping and killing. Recently, it has been implicated in thrombogenesis, autoimmune disease, and cancer progression. The aim of this study was to characterize NETs in various organs of a murine sepsis model in vivo and to investigate their associations with platelets, leukocytes, or vascular endothelium. NETs were classified as two distinct forms; cell-free NETs that were released away from neutrophils and anchored NETs that were anchored to neutrophils. Circulating cell-free NETs were characterized as fragmented or cotton-like structures, while anchored NETs were characterized as linear, reticular, membranous, or spot-like structures. In septic mice, both anchored and cell-free NETs were significantly increased in postcapillary venules of the cecum and hepatic sinusoids with increased leukocyte-endothelial interactions. NETs were also observed in both alveolar space and pulmonary capillaries of the lung. The interactions of NETs with platelet aggregates, leukocyte-platelet aggregates or vascular endothelium of arterioles and venules were observed in the microcirculation of septic mice. Microvessel occlusions which may be caused by platelet aggregates or leukocyte-platelet aggregates and heterogeneously decreased blood flow were also observed in septic mice. NETs appeared to be associated with the formation of platelet aggregates or leukocyte-platelet aggregates. These observational findings may suggest the adverse effect of intravascular NETs on the host during a sepsis.

  8. Novel role of the antimicrobial peptide LL-37 in the protection of neutrophil extracellular traps against degradation by bacterial nucleases

    OpenAIRE

    Neumann, Ariane; Völlger, Lena; Berends, Evelien T.M.; Molhoek, E. Margo; Stapels, Daphne A.C.; Midon, Marika; Friães, Ana; Pingoud, Alfred; Rooijakkers, Suzan H. M.; Richard L Gallo; Mörgelin, Matthias; Nizet, Victor; Naim, Hassan Y; von Köckritz-Blickwede, Maren

    2014-01-01

    Neutrophil extracellular traps (NETs) have been described as a fundamental innate immune defense mechanism. These NETs consist of a nuclear DNA backbone associated with different antimicrobial peptides (AMPs), which are able to engulf and kill pathogens. The AMP LL-37, a member of the cathelicidin family, is highly present in NETs. However, the function of LL-37 within the NETs is still unknown, since LL-37 loses its antimicrobial activity when bound to DNA in the NETs.

  9. Streptococcus suis Serotype 2 Biofilms Inhibit the Formation of Neutrophil Extracellular Traps

    Science.gov (United States)

    Ma, Fang; Yi, Li; Yu, Ningwei; Wang, Guangyu; Ma, Zhe; Lin, Huixing; Fan, Hongjie

    2017-01-01

    Invasive infections caused by Streptococcus suis serotype 2 (SS2) has emerged as a clinical problem in recent years. Neutrophil extracellular traps (NETs) are an important mechanism for the trapping and killing of pathogens that are resistant to phagocytosis. Biofilm formation can protect bacteria from being killed by phagocytes. Until now, there have only been a few studies that focused on the interactions between bacterial biofilms and NETs. SS2 in both a biofilm state and a planktonic cell state were incubated with phagocytes and NETs, and bacterial survival was assessed. DNase I and cytochalasin B were used to degrade NET DNA or suppress phagocytosis, respectively. Extracellular DNA was stained with impermeable fluorescent dye to quantify NET formation. Biofilm formation increased up to 6-fold in the presence of neutrophils, and biofilms were identified in murine tissue. Both planktonic and biofilm cells induced neutrophils chemotaxis to the infection site, with neutrophils increasing by 85.1 and 73.8%, respectively. The bacteria in biofilms were not phagocytized. The bactericidal efficacy of NETs on the biofilms and planktonic cells were equal; however, the biofilm extracellular matrix can inhibit NET release. Although biofilms inhibit NETs release, NETs appear to be an important mechanism to eliminate SS2 biofilms. This knowledge advances the understanding of biofilms and may aid in the development of treatments for persistent infections with a biofilm component. PMID:28373968

  10. Vitamin C: A Novel Regulator of Neutrophil Extracellular Trap Formation

    Directory of Open Access Journals (Sweden)

    Ramesh Natarajan

    2013-08-01

    Full Text Available Introduction: Neutrophil extracellular trap (NET formation was recently identified as a novel mechanism to kill pathogens. However, excessive NET formation in sepsis can injure host tissues. We have recently shown that parenteral vitamin C (VitC is protective in sepsis. Whether VitC alters NETosis is unknown. Methods: We used Gulo−/− mice as they lack the ability to synthesize VitC. Sepsis was induced by intraperitoneal infusion of a fecal stem solution (abdominal peritonitis, FIP. Some VitC deficient Gulo−/− mice received an infusion of ascorbic acid (AscA, 200 mg/kg 30 min after induction of FIP. NETosis was assessed histologically and by quantification for circulating free DNA (cf-DNA in serum. Autophagy, histone citrullination, endoplasmic reticulum (ER stress, NFκB activation and apoptosis were investigated in peritoneal PMNs. Results: Sepsis produced significant NETs in the lungs of VitC deficient Gulo−/− mice and increased circulating cf-DNA. This was attenuated in the VitC sufficient Gulo−/− mice and in VitC deficient Gulo−/− mice infused with AscA. Polymorphonuclear neutrophils (PMNs from VitC deficient Gulo−/− mice demonstrated increased activation of ER stress, autophagy, histone citrullination, and NFκB activation, while apoptosis was inhibited. VitC also significantly attenuated PMA induced NETosis in PMNs from healthy human volunteers.

  11. Nuclease Expression by Staphylococcus aureus Facilitates Escape from Neutrophil Extracellular Traps

    OpenAIRE

    Berends, Evelien T.M.; Horswill, Alexander R.; Haste, Nina M.; Monestier, Marc; Nizet, Victor; von Köckritz-Blickwede, Maren

    2010-01-01

    Neutrophils are key effectors of the host innate immune response against bacterial infection. Staphylococcus aureus is a preeminent human pathogen, with an ability to produce systemic infections even in previously healthy individuals, thereby reflecting a resistance to effective neutrophil clearance. The recent discovery of neutrophil extracellular traps (NETs) has opened a novel dimension in our understanding of how these specialized leukocytes kill pathogens. NETs consist of a nuclear DNA b...

  12. Neutrophil Extracellular Traps and Its Implications in Inflammation: An Overview

    Science.gov (United States)

    Delgado-Rizo, Vidal; Martínez-Guzmán, Marco A.; Iñiguez-Gutierrez, Liliana; García-Orozco, Alejandra; Alvarado-Navarro, Anabell; Fafutis-Morris, Mary

    2017-01-01

    In addition to physical barriers, neutrophils are considered a part of the first line of immune defense. They can be found in the bloodstream, with a lifespan of 6–8 h, and in tissue, where they can last up to 7 days. The mechanisms that neutrophils utilize for host defense are phagocytosis, degranulation, cytokine production, and, the most recently described, neutrophil extracellular trap (NET) production. NETs are DNA structures released due to chromatin decondensation and spreading, and they thus occupy three to five times the volume of condensed chromatin. Several proteins adhere to NETs, including histones and over 30 components of primary and secondary granules, among them components with bactericidal activity such as elastase, myeloperoxidase, cathepsin G, lactoferrin, pentraxin 3, gelatinase, proteinase 3, LL37, peptidoglycan-binding proteins, and others with bactericidal activity able to destroy virulence factors. Three models for NETosis are known to date. (a) Suicidal NETosis, with a duration of 2–4 h, is the best described model. (b) In vital NETosis with nuclear DNA release, neutrophils release NETs without exhibiting loss of nuclear or plasma membrane within 5–60 min, and it is independent of reactive oxygen species (ROS) and the Raf/MERK/ERK pathway. (c) The final type is vital NETosis with release of mitochondrial DNA that is dependent on ROS and produced after stimuli with GM-CSF and lipopolysaccharide. Recent research has revealed neutrophils as more sophisticated immune cells that are able to precisely regulate their granular enzymes release by ion fluxes and can release immunomodulatory cytokines and chemokines that interact with various components of the immune system. Therefore, they can play a key role in autoimmunity and in autoinflammatory and metabolic diseases. In this review, we intend to show the two roles played by neutrophils: as a first line of defense against microorganisms and as a contributor to the pathogenesis of

  13. Ligation of Signal Inhibitory Receptor on Leukocytes-1 Suppresses the Release of Neutrophil Extracellular Traps in Systemic Lupus Erythematosus

    OpenAIRE

    Kristof Van Avondt; Ruth Fritsch-Stork; Derksen, Ronald H W M; Linde Meyaard

    2013-01-01

    Neutrophil extracellular traps (NETs) have been implicated in the pathogenesis of systemic Lupus erythematosus (SLE), since netting neutrophils release potentially immunogenic autoantigens including histones, LL37, human neutrophil peptide (HNP), and self-DNA. In turn, these NETs activate plasmacytoid dendritic cells resulting in aggravation of inflammation and disease. How suppression of NET formation can be targeted for treatment has not been reported yet. Signal Inhibitory Receptor on Leuk...

  14. The role for neutrophil extracellular traps in cystic fibrosis autoimmunity

    Science.gov (United States)

    Skopelja, Sladjana; Hamilton, B. JoNell; Jones, Jonathan D.; Yang, Mei-Ling; Mamula, Mark; Ashare, Alix; Gifford, Alex H.; Rigby, William F.C.

    2016-01-01

    While respiratory failure in cystic fibrosis (CF) frequently associates with chronic infection by Pseudomonas aeruginosa, no single factor predicts the extent of lung damage in CF. To elucidate other causes, we studied the autoantibody profile in CF and rheumatoid arthritis (RA) patients, given the similar association of airway inflammation and autoimmunity in RA. Even though we observed that bactericidal permeability-increasing protein (BPI), carbamylated proteins, and citrullinated proteins all localized to the neutrophil extracellular traps (NETs), which are implicated in the development of autoimmunity, our study demonstrates striking autoantibody specificity in CF. Particularly, CF patients developed anti-BPI autoantibodies but hardly any anti-citrullinated protein autoantibodies (ACPA). In contrast, ACPA-positive RA patients exhibited no reactivity with BPI. Interestingly, anti-carbamylated protein autoantibodies (ACarPA) were found in both cohorts but did not cross-react with BPI. Contrary to ACPA and ACarPA, anti-BPI autoantibodies recognized the BPI C-terminus in the absence of posttranslational modifications. In fact, we discovered that P. aeruginosa–mediated NET formation results in BPI cleavage by P. aeruginosa elastase, which suggests a novel mechanism in the development of autoimmunity to BPI. In accordance with this model, autoantibodies associated with presence of P. aeruginosa on sputum culture. Finally, our results provide a role for autoimmunity in CF disease severity, as autoantibody levels associate with diminished lung function. PMID:27777975

  15. Neutrophil extracellular traps as innate immune reaction against the emerging apicomplexan parasite Besnoitia besnoiti.

    Directory of Open Access Journals (Sweden)

    Tamara Muñoz Caro

    Full Text Available Besnoitia besnoiti infection in cattle is an important emerging protozoan disease in Europe causing economic losses and severe clinical signs, such as generalized dermatitis, orchitis, and vulvitis in affected animals. Neutrophil extracellular trap (NET formation was recently demonstrated as an important effector mechanism of PMN acting against several invading pathogens. In the present study, interactions of bovine PMN with tachyzoites of B. besnoiti were investigated in this respect in vitro. For the demonstration and quantification of NETs, extracellular DNA was stained by Sytox Orange or Pico Green. Fluorescent illustrations as well as scanning electron microscopy analyses (SEM showed PMN-promoted NET formation rapidly being induced upon contact with B. besnoiti tachyzoites. Co-localization of extracellular DNA with histones, neutrophil elastase (NE and myeloperoxidase (MPO in parasite entrapping structures confirmed the classical characteristics of NET. Exposure of PMN to viable, UV attenuated and dead tachyzoites showed a significant induction of NET formation, but even tachyzoite homogenates significantly promoted NETs when compared to negative controls. NETs were abolished by DNase treatment and were reduced after PMN preincubation with NADPH oxidase-, NE- and MPO-inhibitors. Tachyzoite-triggered NET formation led to parasite entrapment as quantitative assays indicated that about one third of tachyzoites were immobilized in NETs. In consequence, tachyzoites were hampered from active invasion of host cells. Thus, transfer of tachyzoites, previously being confronted with PMN, to adequate host cells resulted in significantly reduced infection rates when compared to PMN-free infection controls. To our knowledge, we here report for the first time B. besnoiti-induced NET formation. Our results indicate that PMN-triggered extracellular traps may represent an important effector mechanism of the host early innate immune response against B

  16. Superoxide Induces Neutrophil Extracellular Trap Formation in a TLR-4 and NOX-Dependent Mechanism

    Science.gov (United States)

    Al-Khafaji, Ahmed B; Tohme, Samer; Yazdani, Hamza Obaid; Miller, David; Huang, Hai; Tsung, Allan

    2016-01-01

    Neutrophils constitute the early innate immune response to perceived infectious and sterile threats. Neutrophil extracellular traps (NETs) are a novel mechanism to counter pathogenic invasion and sequelae of ischemia, including cell death and oxidative stress. Superoxide is a radical intermediate of oxygen metabolism produced by parenchymal and nonparenchymal hepatic cells, and is a hallmark of oxidative stress after liver ischemia-reperfusion (I/R). While extracellular superoxide recruits neutrophils to the liver and initiates sterile inflammatory injury, it is unknown whether superoxide induces the formation of NETs. We hypothesize that superoxide induces NET formation through a signaling cascade involving Toll-like receptor 4 (TLR-4) and neutrophil NADPH oxidase (NOX). We treated neutrophils with extracellular superoxide and observed NET DNA release, histone H3 citrullination and increased levels of MPO-DNA complexes occurring in a TLR-4–dependent manner. Inhibition of superoxide generation by allopurinol and inhibition of NOX by diphenyleneiodonium prevented NET formation. When mice were subjected to warm liver I/R, we found significant NET formation associated with liver necrosis and increased serum ALT in TLR-4 WT but not TLR-4 KO mice. To reduce circulating superoxide, we pretreated mice undergoing I/R with allopurinol and N-acetylcysteine, which resulted in decreased NETs and ameliorated liver injury. Our study demonstrates a requirement for TLR-4 and NOX in superoxide-induced NETs, and suggests involvement of superoxide-induced NETs in pathophysiologic settings. PMID:27453505

  17. Neutrophil extracellular traps that are not degraded in systemic lupus erythematosus activate complement exacerbating the disease.

    Science.gov (United States)

    Leffler, Jonatan; Martin, Myriam; Gullstrand, Birgitta; Tydén, Helena; Lood, Christian; Truedsson, Lennart; Bengtsson, Anders A; Blom, Anna M

    2012-04-01

    Ongoing inflammation including activation of the complement system is a hallmark of systemic lupus erythematosus (SLE). Antimicrobial neutrophil extracellular traps (NETs) are composed of secreted chromatin that may act as a source of autoantigens typical for SLE. In this study, we investigated how complement interacts with NETs and how NET degradation is affected by complement in SLE patients. We found that sera from a subset of patients with active SLE had a reduced ability to degrade in vitro-generated NETs, which was mostly restored when these patients were in remission. Patients that failed to degrade NETs had a more active disease and they also displayed lower levels of complement proteins C4 and C3 in blood. We discovered that NETs activated complement in vitro and that deposited C1q inhibited NET degradation including a direct inhibition of DNase-I by C1q. Complement deposition on NETs may facilitate autoantibody production, and indeed, Abs against NETs and NET epitopes were more pronounced in patients with impaired ability to degrade NETs. NET-bound autoantibodies inhibited degradation but also further increased C1q deposition, potentially exacerbating the disease. Thus, NETs are a potent complement activator, and this interaction may play an important role in SLE. Targeting complement with inhibitors or by removing complement activators such as NETs could be beneficial for patients with SLE.

  18. Participation of dectin-1 receptor on NETs release against Paracoccidioides brasiliensis: Role on extracellular killing.

    Science.gov (United States)

    Bachiega, Tatiana Fernanda; Dias-Melicio, Luciane Alarcão; Fernandes, Reginaldo Keller; de Almeida Balderramas, Helanderson; Rodrigues, Daniela Ramos; Ximenes, Valdecir Farias; de Campos Soares, Ângela Maria Victoriano

    2016-02-01

    Paracoccidioides brasiliensis is a dimorphic fungus from the Paracoccidioides genus, which is the causative agent of paracoccidioidomycosis, a chronic, subacute or acute mycosis, with visceral and cutaneous involvement. This disease that is acquired through inhalation primarily attacks the lungs but, can spread to other organs. Phagocytic cells as neutrophils play an important role during innate immune response against this fungus, but studies on antifungal activities of these cells are scarce. In addition to their ability to eliminate pathogens by phagocytosis and antimicrobial secretions, neutrophils can trap and kill microorganisms by release of extracellular structures composed by DNA and antimicrobial proteins, called neutrophil extracellular traps (NETs). Here, we provide evidence that P. brasiliensis virulent strain (P. brasiliensis 18) induces NETs release. These structures were well evidenced by scanning electron microscopy, and specific NETs compounds such as histone, elastase and DNA were shown by confocal microscopy. In addition, we have shown that dectin-1 receptor is the main PRR to which fungus binds to induce NETS release. Fungi were ensnared by NETs, denoting the role of these structures in confining the fungus, avoiding dissemination. NETs were also shown to be involved in fungus killing, since fungicidal activity detected before and mainly after neutrophils activation with TNF-α, IFN-γ and GM-CSF was significantly inhibited by cocultures treatment with DNAse.

  19. Cytokines induced neutrophil extracellular traps formation: implication for the inflammatory disease condition.

    Directory of Open Access Journals (Sweden)

    Ravi S Keshari

    Full Text Available Neutrophils (PMNs and cytokines have a critical role to play in host defense and systemic inflammatory response syndrome (SIRS. Neutrophil extracellular traps (NETs have been shown to extracellularly kill pathogens, and inflammatory potential of NETs has been shown. Microbial killing inside the phagosomes or by NETs is mediated by reactive oxygen and nitrogen species (ROS/RNS. The present study was undertaken to assess circulating NETs contents and frequency of NETs generation by isolated PMNs from SIRS patients. These patients displayed significant augmentation in the circulating myeloperoxidase (MPO activity and DNA content, while PMA stimulated PMNs from these patients, generated more free radicals and NETs. Plasma obtained from SIRS patients, if added to the PMNs isolated from healthy subjects, enhanced NETs release and free radical formation. Expressions of inflammatory cytokines (IL-1β, TNFα and IL-8 in the PMNs as well as their circulating levels were significantly augmented in SIRS subjects. Treatment of neutrophils from healthy subjects with TNFα, IL-1β, or IL-8 enhanced free radicals generation and NETs formation, which was mediated through the activation of NADPH oxidase and MPO. Pre-incubation of plasma from SIRS with TNFα, IL-1β, or IL-8 antibodies reduced the NETs release. Role of IL-1β, TNFα and IL-8 thus seems to be involved in the enhanced release of NETs in SIRS subjects.

  20. An improved strategy to recover large fragments of functional human neutrophil extracellular traps

    Directory of Open Access Journals (Sweden)

    Lorena eBarrientos

    2013-06-01

    Full Text Available Netosis is a recently described neutrophil function that leads to the release of neutrophil extracellular traps (NETs in response to various stimuli. NETs are filaments of decondensed chromatin associated with granular proteins. In addition to their role against microorganisms, NETs have been implicated in autoimmunity, thrombosis and tissue injury. Access to a standardized source of isolated NETs is needed to better analyze the roles of NETs. The aim of this study was to develop a procedure yielding soluble, well-characterized NET preparations from fresh human neutrophils. The calcium ionophore A23187 was chosen to induce netosis, and the restriction enzyme Alu I was used to prepare large NET fragments. DNA and proteins were detected by electrophoresis and specific labeling. Some NET proteins (histone 3, lactoferrin were quantified by western blotting, and dsDNA was quantified by immunofluorescence. Co-existence of dsDNA and neutrophil proteins confirmed the quality of the NET preparations. Their biological activity was checked by measuring elastase activity and bacterial killing against various strains. Interindividual differences in histone 3, lactoferrin, elastase and dsDNA relative contents were observed in isolated NETs. However, the reproducibility of NET preparation and characterization was validated, suggesting that this interindividual variability was rather related to donor variation than to technical bias. This standardized protocol is suitable for producing, isolating and quantifying functional NETs that could serve as a tool for studying NET effects on immune cells and tissues.

  1. An improved strategy to recover large fragments of functional human neutrophil extracellular traps.

    Science.gov (United States)

    Barrientos, Lorena; Marin-Esteban, Viviana; de Chaisemartin, Luc; Le-Moal, Vanessa Lievin; Sandré, Catherine; Bianchini, Elsa; Nicolas, Valerie; Pallardy, Marc; Chollet-Martin, Sylvie

    2013-01-01

    Netosis is a recently described neutrophil function that leads to the release of neutrophil extracellular traps (NETs) in response to various stimuli. NETs are filaments of decondensed chromatin associated with granular proteins. In addition to their role against microorganisms, NETs have been implicated in autoimmunity, thrombosis, and tissue injury. Access to a standardized source of isolated NETs is needed to better analyze the roles of NETs. The aim of this study was to develop a procedure yielding soluble, well-characterized NET preparations from fresh human neutrophils. The calcium ionophore A23187 was chosen to induce netosis, and the restriction enzyme AluI was used to prepare large NET fragments. DNA and proteins were detected by electrophoresis and specific labeling. Some NET proteins [histone 3, lactoferrin (LF)] were quantified by western blotting, and double-stranded DNA (dsDNA) was quantified by immunofluorescence. Co-existence of dsDNA and neutrophil proteins confirmed the quality of the NET preparations. Their biological activity was checked by measuring elastase (ELA) activity and bacterial killing against various strains. Interindividual differences in histone 3, LF, ELA, and dsDNA relative contents were observed in isolated NETs. However, the reproducibility of NET preparation and characterization was validated, suggesting that this interindividual variability was rather related to donor variation than to technical bias. This standardized protocol is suitable for producing, isolating, and quantifying functional NETs that could serve as a tool for studying NET effects on immune cells and tissues.

  2. Neutrophil extracellular traps directly induce epithelial and endothelial cell death: a predominant role of histones.

    Directory of Open Access Journals (Sweden)

    Mona Saffarzadeh

    Full Text Available Neutrophils play an important role in innate immunity by defending the host organism against invading microorganisms. Antimicrobial activity of neutrophils is mediated by release of antimicrobial peptides, phagocytosis as well as formation of neutrophil extracellular traps (NET. These structures are composed of DNA, histones and granular proteins such as neutrophil elastase and myeloperoxidase. This study focused on the influence of NET on the host cell functions, particularly on human alveolar epithelial cells as the major cells responsible for gas exchange in the lung. Upon direct interaction with epithelial and endothelial cells, NET induced cytotoxic effects in a dose-dependent manner, and digestion of DNA in NET did not change NET-mediated cytotoxicity. Pre-incubation of NET with antibodies against histones, with polysialic acid or with myeloperoxidase inhibitor but not with elastase inhibitor reduced NET-mediated cytotoxicity, suggesting that histones and myeloperoxidase are responsible for NET-mediated cytotoxicity. Although activated protein C (APC did decrease the histone-induced cytotoxicity in a purified system, it did not change NET-induced cytotoxicity, indicating that histone-dependent cytotoxicity of NET is protected against APC degradation. Moreover, in LPS-induced acute lung injury mouse model, NET formation was documented in the lung tissue as well as in the bronchoalveolar lavage fluid. These data reveal the important role of protein components in NET, particularly histones, which may lead to host cell cytotoxicity and may be involved in lung tissue destruction.

  3. Neutrophil extracellular traps contain calprotectin, a cytosolic protein complex involved in host defense against Candida albicans.

    Directory of Open Access Journals (Sweden)

    Constantin F Urban

    2009-10-01

    Full Text Available Neutrophils are the first line of defense at the site of an infection. They encounter and kill microbes intracellularly upon phagocytosis or extracellularly by degranulation of antimicrobial proteins and the release of Neutrophil Extracellular Traps (NETs. NETs were shown to ensnare and kill microbes. However, their complete protein composition and the antimicrobial mechanism are not well understood. Using a proteomic approach, we identified 24 NET-associated proteins. Quantitative analysis of these proteins and high resolution electron microscopy showed that NETs consist of modified nucleosomes and a stringent selection of other proteins. In contrast to previous results, we found several NET proteins that are cytoplasmic in unstimulated neutrophils. We demonstrated that of those proteins, the antimicrobial heterodimer calprotectin is released in NETs as the major antifungal component. Absence of calprotectin in NETs resulted in complete loss of antifungal activity in vitro. Analysis of three different Candida albicans in vivo infection models indicated that NET formation is a hitherto unrecognized route of calprotectin release. By comparing wild-type and calprotectin-deficient animals we found that calprotectin is crucial for the clearance of infection. Taken together, the present investigations confirmed the antifungal activity of calprotectin in vitro and, moreover, demonstrated that it contributes to effective host defense against C. albicans in vivo. We showed for the first time that a proportion of calprotectin is bound to NETs in vitro and in vivo.

  4. Neutrophil extracellular traps contain calprotectin, a cytosolic protein complex involved in host defense against Candida albicans.

    Science.gov (United States)

    Urban, Constantin F; Ermert, David; Schmid, Monika; Abu-Abed, Ulrike; Goosmann, Christian; Nacken, Wolfgang; Brinkmann, Volker; Jungblut, Peter R; Zychlinsky, Arturo

    2009-10-01

    Neutrophils are the first line of defense at the site of an infection. They encounter and kill microbes intracellularly upon phagocytosis or extracellularly by degranulation of antimicrobial proteins and the release of Neutrophil Extracellular Traps (NETs). NETs were shown to ensnare and kill microbes. However, their complete protein composition and the antimicrobial mechanism are not well understood. Using a proteomic approach, we identified 24 NET-associated proteins. Quantitative analysis of these proteins and high resolution electron microscopy showed that NETs consist of modified nucleosomes and a stringent selection of other proteins. In contrast to previous results, we found several NET proteins that are cytoplasmic in unstimulated neutrophils. We demonstrated that of those proteins, the antimicrobial heterodimer calprotectin is released in NETs as the major antifungal component. Absence of calprotectin in NETs resulted in complete loss of antifungal activity in vitro. Analysis of three different Candida albicans in vivo infection models indicated that NET formation is a hitherto unrecognized route of calprotectin release. By comparing wild-type and calprotectin-deficient animals we found that calprotectin is crucial for the clearance of infection. Taken together, the present investigations confirmed the antifungal activity of calprotectin in vitro and, moreover, demonstrated that it contributes to effective host defense against C. albicans in vivo. We showed for the first time that a proportion of calprotectin is bound to NETs in vitro and in vivo.

  5. Extracellular traps are associated with human and mouse neutrophil and macrophage mediated killing of larval Strongyloides stercoralis.

    Science.gov (United States)

    Bonne-Année, Sandra; Kerepesi, Laura A; Hess, Jessica A; Wesolowski, Jordan; Paumet, Fabienne; Lok, James B; Nolan, Thomas J; Abraham, David

    2014-06-01

    Neutrophils are multifaceted cells that are often the immune system's first line of defense. Human and murine cells release extracellular DNA traps (ETs) in response to several pathogens and diseases. Neutrophil extracellular trap (NET) formation is crucial to trapping and killing extracellular pathogens. Aside from neutrophils, macrophages and eosinophils also release ETs. We hypothesized that ETs serve as a mechanism of ensnaring the large and highly motile helminth parasite Strongyloides stercoralis thereby providing a static target for the immune response. We demonstrated that S. stercoralis larvae trigger the release of ETs by human neutrophils and macrophages. Analysis of NETs revealed that NETs trapped but did not kill larvae. Induction of NETs was essential for larval killing by human but not murine neutrophils and macrophages in vitro. In mice, extracellular traps were induced following infection with S. stercoralis larvae and were present in the microenvironment of worms being killed in vivo. These findings demonstrate that NETs ensnare the parasite facilitating larval killing by cells of the immune system.

  6. The Protective Effect against Extracellular Histones Afforded by Long-Pentraxin PTX3 as a Regulator of NETs

    Science.gov (United States)

    Daigo, Kenji; Takamatsu, Yuichiro; Hamakubo, Takao

    2016-01-01

    Pentraxin 3 (PTX3) is a soluble pattern recognition molecule that plays critical roles in innate immunity. Its fundamental functions include recognition of microbes, activation of complement cascades, and opsonization. The findings that PTX3 is one of the component proteins in neutrophil extracellular traps (NETs) and binds with other NET proteins imply the importance of PTX3 in the NET-mediated trapping and killing of bacteria. As NETs play certain critically important host-protective roles, aberrant NET production results in tissue damage. Extracellular histones, the main source of which is considered to be NETs, are mediators of septic death due to their cytotoxicity toward endothelial cells. PTX3 protects against extracellular histones-mediated cytotoxicity through coaggregation. In addition to the anti-bacterial roles performed in coordination with other NET proteins, PTX3 appears to mitigate the detrimental effect of over-activated NETs. A better understanding of the role of the PTX3 complexes in NETs would be expected to lead to new strategies for maintaining a healthy balance between the helpful bactericidal and undesirable detrimental activities of NETs. PMID:27656184

  7. DNase expression allows the pathogen group A streptococcus to escape killing in neutrophil extracellular traps

    OpenAIRE

    Buchanan, John T; Simpson, Amelia J; Aziz, Ramy K.; Liu, George Y.; Kristian, Sascha A.; Kotb, Malak; Feramisco, James; Nizet, Victor

    2006-01-01

    The innate immune response plays a crucial role in satisfactory host resolution of bacterial infection. In response to chemotactic signals, neutrophils are early responding cells that migrate in large numbers to sites of infection. The recent discovery of secreted neutrophil extracellular traps (NETs) composed of DNA and histones opened a novel dimension in our understanding of the microbial killing capacity of these specialized leukocytes. M1 serotype strains of the pathogen Group A Streptoc...

  8. Neutrophil extracellular trap cell death requires both autophagy and superoxide generation

    Institute of Scientific and Technical Information of China (English)

    Quinten Remijsen; Peter Vandenabeele; Tom Vanden Berghe; Ellen Wirawan; Bob Asselbergh; Eef Parthoens; Riet De Rycke; Sam Noppen; Michel Delforge; Jean Willems

    2011-01-01

    Neutrophil extracellular traps(NETs)are extracellular chromatin structures that can trap and degrade microbes.They arise from neutrophils that have activated a cell death program called NET cell death,or NETosis.Activation of NETosis has been shown to involve NADPH oxidase activity,disintegration of the nuclear envelope and most granule membranes,decondensation of nuclear chromatin and formation of NETs.We report that in phorbol myristate acetate(PMA)-stimulated neutrophils,intracellular chromatin decondensation and NET formation follow autophagy and superoxide production,both of which arerequired to mediate PMA-induced NETosis and occur independently of each other.Neutrophils from patients with chronic granulomatous disease,which lack NADPH oxidase activity,still exhibit PMA-induced autophagy.Conversely,PMA-induced NADPH oxidase activity is not affected by pharmacological inhibition of autophagy.Interestingly,inhibition of either autophagy or NADPH oxidase prevents intracellular chromatin decondensation,which is essential for NETosis and NET formation,and results in cell death characterized by hallmarks of apoptosis.These results indicate that apoptosis might function as a backup program for NETosis when autophagy or NADPH oxidase activity is prevented.

  9. Carp neutrophilic granulocytes form extracellular traps via ROS-dependent and independent pathways.

    Science.gov (United States)

    Pijanowski, L; Golbach, L; Kolaczkowska, E; Scheer, M; Verburg-van Kemenade, B M L; Chadzinska, M

    2013-05-01

    Neutrophil extracellular traps (NETs) have recently been described as an important innate defense mechanism that leads to immobilization and killing of invading pathogens. NETs have been identified in several species, but the mechanisms involved in NET formation and their role in infection have not been well determined yet. Here we show that upon in vitro stimulation with different immunostimulants of bacterial, fungal or viral origin, carp neutrophilic granulocytes rapidly release NET structures. We analyzed the composition of these structures and the kinetics of their formation by confocal microscopy, by quantifying the levels of extracellular DNA and the release of enzymes originating from neutrophilic granules: myeloperoxidase, neutrophil elastase and matrix metalloproteinase 9 (MMP-9). Profiles of NET release by carp neutrophils as well as their enzyme composition are stimulus- and time-dependent. This study moreover provides evidence for a stimulus-dependent selective requirement of reactive oxygen species in the process of NET formation. Collectively the results support an evolutionary conserved and strictly regulated mechanism of NET formation in teleost fish.

  10. Are Neutrophil Extracellular Traps Playing a Role in the Parasite Control in Active American Tegumentary Leishmaniasis Lesions?

    Science.gov (United States)

    Morgado, Fernanda Nazaré; Nascimento, Michelle T C; Saraiva, Elvira M; de Oliveira-Ribeiro, Carla; Madeira, Maria de Fátima; da Costa-Santos, Marcela; Vasconcellos, Erica C F; Pimentel, Maria Ines F; Rosandiski Lyra, Marcelo; Schubach, Armando de Oliveira; Conceição-Silva, Fátima

    2015-01-01

    Neutrophil extracellular traps (NETs) have been described as a network of extracellular fibers composed by DNA, histones and various proteins/enzymes. Studies have demonstrated that NETs could be responsible for the trapping and elimination of a variety of infectious agents. In order to verify the presence of NETs in American tegumentary leishmaniasis (ATL) and their relationship with the presence of amastigotes we evaluated active cutaneous lesions of 35 patients before treatment by the detection of parasites, neutrophils (neutrophil elastase) and histones through immunohistochemistry and confocal immunofluorescence. Intact neutrophils could be detected in all ATL lesions. NETs were present in 27 patients (median 1.1; range from 0.1 to 23.5/mm2) with lesion duration ranging from one to seven months. NETs were in close proximity with neutrophils (r = 0.586; p = 0.0001) and amastigotes (r = 0.710; p = 0.0001). Two patterns of NET formation were detected: small homogeneously distributed networks observed in all lesions; and large structures that could be visualized at a lower magnification in lesions presenting at least 20% of neutrophils. Lesions presenting the larger NET formation showed high parasite detection. A correlation between NET size and the number of intact amastigotes was observed (p=0.02). As we detected an association between NET and amastigotes, our results suggest that neutrophil migration and NET formation could be stimulated and maintained by stimuli derived from the parasite burden/parasite antigen in the extracellular environment. The observation of areas containing only antigens not intermingled with NETs (elastase and histone) suggests that the involvement of these structures in the control of parasite burden is a dynamic process in which the formation of NETs is exhausted with the destruction of the parasites. Since NETs were also associated with granulomas, this trapping would favor the activity of macrophages in order to control the parasite

  11. Neutrophils recruited to sites of infection protect from virus challenge by releasing neutrophil extracellular traps.

    Science.gov (United States)

    Jenne, Craig N; Wong, Connie H Y; Zemp, Franz J; McDonald, Braedon; Rahman, Masmudur M; Forsyth, Peter A; McFadden, Grant; Kubes, Paul

    2013-02-13

    Neutrophils mediate bacterial clearance through various mechanisms, including the release of mesh-like DNA structures or neutrophil extracellular traps (NETs) that capture bacteria. Although neutrophils are also recruited to sites of viral infection, their role in antiviral innate immunity is less clear. We show that systemic administration of virus analogs or poxvirus infection induces neutrophil recruitment to the liver microvasculature and the release of NETs that protect host cells from virus infection. After systemic intravenous poxvirus challenge, mice exhibit thrombocytopenia and the recruitment of both neutrophils and platelets to the liver vasculature. Circulating platelets interact with, roll along, and adhere to the surface of adherent neutrophils, forming large, dynamic aggregates. These interactions facilitate the release of NETs within the liver vasculature that are able to protect host cells from poxvirus infection. These findings highlight the role of NETs and early tissue-wide responses in preventing viral infection.

  12. Do neutrophil extracellular traps contribute to the heightened risk of thrombosis in inflammatory diseases?

    Institute of Scientific and Technical Information of China (English)

    Ashish N Rao; Nayef M Kazzaz; Jason S Knight

    2015-01-01

    Thrombotic events,both arterial and venous,are a major health concern worldwide. Further,autoimmune diseases,such as systemic lupus erythematosus,anti-neutrophil cytoplasmic antibody(ANCA)-associated vasculitis,and antiphospholipid syndrome,predispose to thrombosis,and thereby push the risk for these morbid events even higher. In recent years,neutrophils have been identified as important players in both arterial and venous thrombosis. Specifically,chromatin-based structures called neutrophil extracellular traps(NETs) play a key role in activating the coagulation cascade,recruiting platelets,and serving as scaffolding upon which the thrombus can be assembled. At the same time,neutrophils and NETs are emerging as important mediators of pathogenic inflammation in the aforementioned autoimmune diseases. Here,we first review the general role of NETs in thrombosis. We then posit that exaggerated NET release contributes to the prothrombotic diatheses of systemic lupus erythematosus,ANCA-associated vasculitis,and antiphospholipid syndrome.

  13. Abundant neutrophil extracellular traps in thrombus of patient with microscopic polyangiitis

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    Daigo eNakazawa

    2012-11-01

    Full Text Available This is a case study of a patient diagnosed with microscopic polyangiitis (MPA and complicated with deep vein thrombosis (DVT, who died of respiratory failure despite treatment. Autopsy revealed severe crescentic glomerulonephritis and massive alveolar hemorrhage. The thrombus contained abundant neutrophils. Although it is reported that patients with ANCA-associated vasculitis (AAV have an increased risk of DVT, it remains elusive why they are prone to thrombosis. A recent study has demonstrated the presence of neutrophil extracellular traps (NETs, a newly recognized mode of neutrophil cell-death, in glomerular crescents of MPA patients. Interestingly, NETs were identified in the thrombus as well as in the glomerular crescents in the present case. When compared to other thrombi unrelated to MPA, the amount of NETs was significantly greater in the MPA patient. On the other hand, NETs are critically involved in thrombogenesis because histones within NETs can bind platelets and blood coagulants. Although this is important in regard to containment of microbes within NETs, excessive NETs could cause thrombosis. The collective findings suggest the possibility that thrombosis could be critically associated with MPA via NETs, and that NETs could be a therapeutic target in MPA patients.

  14. Respiratory syncytial virus fusion protein promotes TLR-4-dependent neutrophil extracellular trap formation by human neutrophils.

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    Giselle A Funchal

    Full Text Available Acute viral bronchiolitis by Respiratory Syncytial Virus (RSV is the most common respiratory illness in children in the first year of life. RSV bronchiolitis generates large numbers of hospitalizations and an important burden to health systems. Neutrophils and their products are present in the airways of RSV-infected patients who developed increased lung disease. Neutrophil Extracellular Traps (NETs are formed by the release of granular and nuclear contents of neutrophils in the extracellular space in response to different stimuli and recent studies have proposed a role for NETs in viral infections. In this study, we show that RSV particles and RSV Fusion protein were both capable of inducing NET formation by human neutrophils. Moreover, we analyzed the mechanisms involved in RSV Fusion protein-induced NET formation. RSV F protein was able to induce NET release in a concentration-dependent fashion with both neutrophil elastase and myeloperoxidase expressed on DNA fibers and F protein-induced NETs was dismantled by DNase treatment, confirming that their backbone is chromatin. This viral protein caused the release of extracellular DNA dependent on TLR-4 activation, NADPH Oxidase-derived ROS production and ERK and p38 MAPK phosphorylation. Together, these results demonstrate a coordinated signaling pathway activated by F protein that led to NET production. The massive production of NETs in RSV infection could aggravate the inflammatory symptoms of the infection in young children and babies. We propose that targeting the binding of TLR-4 by F protein could potentially lead to novel therapeutic approaches to help control RSV-induced inflammatory consequences and pathology of viral bronchiolitis.

  15. Neutrophils extracellular traps damage Naegleria fowleri trophozoites opsonized with human IgG.

    Science.gov (United States)

    Contis-Montes de Oca, A; Carrasco-Yépez, M; Campos-Rodríguez, R; Pacheco-Yépez, J; Bonilla-Lemus, P; Pérez-López, J; Rojas-Hernández, S

    2016-08-01

    Naegleria fowleri infects humans through the nasal mucosa causing a disease in the central nervous system known as primary amoebic meningoencephalitis (PAM). Polymorphonuclear cells (PMNs) play a critical role in the early phase of N. fowleri infection. Recently, a new biological defence mechanism called neutrophil extracellular traps (NETs) has been attracting attention. NETs are composed of nuclear DNA combined with histones and antibacterial proteins, and these structures are released from the cell to direct its antimicrobial attack. In this work, we evaluate the capacity of N. fowleri to induce the liberation of NETs by human PMN cells. Neutrophils were cocultured with unopsonized or IgG-opsonized N. fowleri trophozoites. DNA, histone, myeloperoxidase (MPO) and neutrophil elastase (NE) were stained, and the formation of NETs was evaluated by confocal microscopy and by quantifying the levels of extracellular DNA. Our results showed N. fowleri induce the liberation of NETs including release of MPO and NE by human PMN cells as exposure interaction time is increased, but N. fowleri trophozoites evaded killing. However, when trophozoites were opsonized, they were susceptible to the neutrophils activity. Therefore, our study suggests that antibody-mediated PMNs activation through NET formation may be crucial for antimicrobial responses against N. fowleri.

  16. Neutrophil extracellular traps in the intestinal mucosa of Eimeria-infected animals

    Institute of Scientific and Technical Information of China (English)

    Tamara Mu?oz-Caro; Liliana Machado Ribeiro da Silva; Zaída Rentería-Solis; Anja Taubert; Carlos Hermosilla

    2016-01-01

    Objective:To investigate the presence of neutrophil extracellular traps(NETs) in vivo by analysing intestinal sections from experimentally Eimeria bovis-and naturally Eimeria arloingi-infected animals.Methods:Intestinal samples of Eimeria arloingi-and Eimeria bovis-infected animals were analysed by using immunohistochemical and fluorescence approach by using monoclonal antibodies.Results:Classical NET components were confirmed by co-localization of extracellular DNA being decorated with neutrophil elastase and histones in Eimeria-infected tissue samples.Here,extrusion of NETs was exclusively detected in intestinal polymorphonuclear neutrophils infiltrating Eimeria-infected sites.In vivo NETs were either found in close proximity or in direct contact to different Eimeria stages suggesting a stage-independent process.NETs were also found within the gut lumen driven by polymorphonuclear neutrophils that were contacting released oocysts.Conclusions:We postulate that NETs might play an important role in innate defence reactions in coccidiosis therefore significantly altering the outcome of infection.

  17. Pyocyanin-enhanced neutrophil extracellular trap formation requires the NADPH oxidase.

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    Balázs Rada

    Full Text Available Beyond intracellular killing, a novel neutrophil-based antimicrobial mechanism has been recently discovered: entrapment and killing by neutrophil extracellular traps (NETs. NETs consist of extruded nuclear DNA webs decorated with granule proteins. Although NET formation is an important innate immune mechanism, uncontrolled NET release damages host tissues and has been linked to several diseases including cystic fibrosis (CF. The major CF airway pathogen Pseudomonas aeruginosa establishes chronic infection. Pseudomonas imbedded within biofilms is protected against the immune system, but maintains chronic inflammation that worsens disease symptoms. Aberrant NET release from recruited neutrophils was found in CF, but the underlying mechanisms remain unclear. One of the most important Pseudomonas virulence factors is pyocyanin, a redox-active pigment that has been associated with diminished lung function in CF. Here we show that pyocyanin promotes NET formation in a time- and dose-dependent manner. Most CF Pseudomonas clinical isolates tested produce pyocyanin in vitro. Pyocyanin-derived reactive oxygen species are required for its NET release. Inhibitor experiments demonstrated involvement of Jun N-terminal Kinase (JNK and phosphatidylinositol 3-Kinase (PI3K in pyocyanin-induced NET formation. Pyocyanin-induced NETs also require the NADPH oxidase because NET release in chronic granulomatous disease neutrophils was greatly reduced. Comparison of neutrophils from gp91phox- and p47phox-deficient patients revealed that pyocyanin-triggered NET formation is proportional to their residual superoxide production. Our studies identify pyocyanin as the first secreted bacterial toxin that enhances NET formation. The involvement of NADPH oxidase in pyocyanin-induced NET formation represents a novel mechanism of pyocyanin toxicity.

  18. Canine Neutrophil Extracellular Traps Release Induced by the Apicomplexan Parasite Neospora caninum In Vitro

    Science.gov (United States)

    Wei, Zhengkai; Hermosilla, Carlos; Taubert, Anja; He, Xuexiu; Wang, Xiaocen; Gong, Pengtao; Li, Jianhua; Yang, Zhengtao; Zhang, Xichen

    2016-01-01

    Neosporosis is considered as one of the main causes of abortion and severe economic losses in dairy industry. The Canis genus serving as one of the confirmed definitive hosts of the apicomplexan parasite Neospora caninum (N. caninum) plays a critical role in its life cycle. However, the effects of N. caninum on its definitive hosts of neutrophils extracellular traps (NETs) formation remain unclear. In the present study, N. caninum tachyzoite-induced canine NETs formation was observed by scanning electron microscopy (SEM). Visualization of DNA decorated with H3, neutrophil elastase (NE), and myeloperoxidase (MPO) within N. caninum tachyzoite-induced NETs were examined using fluorescence confocal microscopy analyses. Furthermore, the formation of canine NETs was quantified using Sytox Green staining, and the LDH levels in supernatants were examined by an LDH Cytotoxicity Assay® kit. The results clearly showed that NETs-like structures were induced by N. caninum tachyzoites, and the major components within these structures induced by N. caninum tachyzoite were further confirmed by fluorescence confocal microscopy visualization. These results suggest that N. caninum tachyzoites strongly induced NETs formation in canine polymorphonuclear neutrophils (PMN). In functional inhibition assays, the blockings of NADPH oxidase, NE, MPO, SOCE, ERK 1/2, and p38 MAPK signaling pathways significantly inhibited N. caninum tachyzoite-induced NETs formation. To our knowledge, this study is the first to report the formation of NETs in canine PMN against N. caninum infection. PMID:27843440

  19. Canine Neutrophil Extracellular Traps Release Induced by the Apicomplexan Parasite Neospora caninum In Vitro.

    Science.gov (United States)

    Wei, Zhengkai; Hermosilla, Carlos; Taubert, Anja; He, Xuexiu; Wang, Xiaocen; Gong, Pengtao; Li, Jianhua; Yang, Zhengtao; Zhang, Xichen

    2016-01-01

    Neosporosis is considered as one of the main causes of abortion and severe economic losses in dairy industry. The Canis genus serving as one of the confirmed definitive hosts of the apicomplexan parasite Neospora caninum (N. caninum) plays a critical role in its life cycle. However, the effects of N. caninum on its definitive hosts of neutrophils extracellular traps (NETs) formation remain unclear. In the present study, N. caninum tachyzoite-induced canine NETs formation was observed by scanning electron microscopy (SEM). Visualization of DNA decorated with H3, neutrophil elastase (NE), and myeloperoxidase (MPO) within N. caninum tachyzoite-induced NETs were examined using fluorescence confocal microscopy analyses. Furthermore, the formation of canine NETs was quantified using Sytox Green staining, and the LDH levels in supernatants were examined by an LDH Cytotoxicity Assay(®) kit. The results clearly showed that NETs-like structures were induced by N. caninum tachyzoites, and the major components within these structures induced by N. caninum tachyzoite were further confirmed by fluorescence confocal microscopy visualization. These results suggest that N. caninum tachyzoites strongly induced NETs formation in canine polymorphonuclear neutrophils (PMN). In functional inhibition assays, the blockings of NADPH oxidase, NE, MPO, SOCE, ERK 1/2, and p38 MAPK signaling pathways significantly inhibited N. caninum tachyzoite-induced NETs formation. To our knowledge, this study is the first to report the formation of NETs in canine PMN against N. caninum infection.

  20. Different virulence of candida albicans is attributed to the ability of escape from neutrophil extracellular traps by secretion of DNase

    Science.gov (United States)

    Zhang, Xiaohuan; Zhao, Sainan; Sun, Luping; Li, Wenqing; Wei, Qiao; Ashman, Robert B; Hu, Yan

    2017-01-01

    Candida albicans is an important opportunistic fungus causing both disseminated and local infections. The discovery of neutrophil extracellular traps (NETs) has presented a new strategy to kill microorganisms in host’s innate immune response. Although it has been reported that NETs can trap and kill both yeast and hyphal forms of C. albicans, the mechanism by which C. albicans escape from NETs has not been fully understood. In this study, the ability of two strains of C. albicans SC5314 and 3683 to escape NETs-mediated killing was compared. It was found that SC5314 induced higher levels of reactive oxygen species (ROS) and expressions of Rac1/2 and more NETs formation by neutrophils, and also generated more deoxyribonucleases (DNase) than 3683 did. However, resistance to neutrophils killing was greater in SC5314 than that of 3683. When extracellular traps were degraded by exogenous DNase I or catalase, and neutrophil phagocytic activity blocked by cytochalasin D, the killing capacity of neutrophils co-cultured with either C. albicans SC5314 or 3683 was significantly decreased. This study indicates that C. albicans can escape from the trapping and killing of NETs by secreting DNase, which offers further insights into the basis for differences in virulence of different strains of C. albicans. PMID:28123633

  1. Neutrophil extracellular traps form a barrier between necrotic and viable areas in acute abdominal inflammation

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    Rostyslav Bilyy

    2016-10-01

    Full Text Available Neutrophils form neutrophil extracellular traps (NETs of decondensed DNA and histones that trap and immobilize particulate matter and microbial pathogens like bacteria. NET aggregates reportedly surround and isolate large objects like monosodium urate (MSU crystals, which cannot be sufficiently cleared from tissues. In the setting of acute necrotizing pancreatitis massive tissue necrosis occurs, which is organized as pancreatic pseudocysts. In contrast to regular cysts, these pseudocysts are not surrounded by epithelial layers. We hypothesize that, instead, the necrotic areas observed in necrotizing pancreatitis are isolated from the surrounding healthy tissues by aggregated NETs. These may form an alternative, putatively transient barrier separating necrotic areas from viable tissue. To test this hypothesis, we investigated histological samples from the necropsy material of internal organs of two patients with necrotizing pancreatitis and peritonitis accompanied by multiple organ failure. Tissues including the inflammatory zone were stained with H&E and evaluated for signs of inflammation. Infiltrating neutrophils and NETs were detected by immunohistochemistry for DNA, neutrophil elastase, and citrullinated histone H3. Interestingly, in severely affected areas of pancreatic necrosis or peritonitis, chromatin stained positive for neutrophil elastase and citrullinated histone H3, and may, therefore, be considered NET-derived. These NET structures formed a layer which separated the necrotic core from the areas of viable tissue remains. A condensed layer of aggregated NETs thus spatially shields and isolates the site of necrosis, thereby limiting the spread of necrosis-associated proinflammatory mediators. We propose that necrotic debris may initiate and/or facilitate the formation of the NET-based surrogate barrier.

  2. Identification and characterization of neutrophil extracellular trap shapes in flow cytometry

    Science.gov (United States)

    Ginley, Brandon; Emmons, Tiffany; Sasankan, Prabhu; Urban, Constantin; Segal, Brahm H.; Sarder, Pinaki

    2017-03-01

    Neutrophil extracellular trap (NET) formation is an alternate immunologic weapon used mainly by neutrophils. Chromatin backbones fused with proteins derived from granules are shot like projectiles onto foreign invaders. It is thought that this mechanism is highly anti-microbial, aids in preventing bacterial dissemination, is used to break down structures several sizes larger than neutrophils themselves, and may have several more uses yet unknown. NETs have been implied to be involved in a wide array of systemic host immune defenses, including sepsis, autoimmune diseases, and cancer. Existing methods used to visually quantify NETotic versus non-NETotic shapes are extremely time-consuming and subject to user bias. These limitations are obstacles to developing NETs as prognostic biomarkers and therapeutic targets. We propose an automated pipeline for quantitatively detecting neutrophil and NET shapes captured using a flow cytometry-imaging system. Our method uses contrast limited adaptive histogram equalization to improve signal intensity in dimly illuminated NETs. From the contrast improved image, fixed value thresholding is applied to convert the image to binary. Feature extraction is performed on the resulting binary image, by calculating region properties of the resulting foreground structures. Classification of the resulting features is performed using Support Vector Machine. Our method classifies NETs from neutrophils without traps at 0.97/0.96 sensitivity/specificity on n = 387 images, and is 1500X faster than manual classification, per sample. Our method can be extended to rapidly analyze whole-slide immunofluorescence tissue images for NET classification, and has potential to streamline the quantification of NETs for patients with diseases associated with cancer and autoimmunity.

  3. Signal Inhibitory Receptor on Leukocytes-1 Limits the Formation of Neutrophil Extracellular Traps, but Preserves Intracellular Bacterial Killing.

    Science.gov (United States)

    Van Avondt, Kristof; van der Linden, Maarten; Naccache, Paul H; Egan, David A; Meyaard, Linde

    2016-05-01

    In response to microbial invasion, neutrophils release neutrophil extracellular traps (NETs) to trap and kill extracellular microbes. Alternatively, NET formation can result in tissue damage in inflammatory conditions and may perpetuate autoimmune disease. Intervention strategies that are aimed at modifying pathogenic NET formation should ideally preserve other neutrophil antimicrobial functions. We now show that signal inhibitory receptor on leukocytes-1 (SIRL-1) attenuates NET release by human neutrophils in response to distinct triggers, including opsonized Staphylococcus aureus and inflammatory danger signals. NET release has different kinetics depending on the stimulus, and rapid NET formation is independent of NADPH oxidase activity. In line with this, we show that NET release and reactive oxygen species production upon challenge with opsonized S. aureus require different signaling events. Importantly, engagement of SIRL-1 does not affect bacterially induced production of reactive oxygen species, and intracellular bacterial killing by neutrophils remains intact. Thus, our studies define SIRL-1 as an intervention point of benefit to suppress NET formation in disease while preserving intracellular antimicrobial defense.

  4. Characterization of Early-Phase Neutrophil Extracellular Traps in Urinary Tract Infections.

    Science.gov (United States)

    Yu, Yanbao; Kwon, Keehwan; Tsitrin, Tamara; Bekele, Shiferaw; Sikorski, Patricia; Nelson, Karen E; Pieper, Rembert

    2017-01-01

    Neutrophils have an important role in the antimicrobial defense and resolution of urinary tract infections (UTIs). Our research suggests that a mechanism known as neutrophil extracellular trap (NET) formation is a defense strategy to combat pathogens that have invaded the urinary tract. A set of human urine specimens with very high neutrophil counts had microscopic evidence of cellular aggregation and lysis. Deoxyribonuclease I (DNase) treatment resulted in disaggregation of such structures, release of DNA fragments and a proteome enriched in histones and azurophilic granule effectors whose quantitative composition was similar to that of previously described in vitro-formed NETs. The effector proteins were further enriched in DNA-protein complexes isolated in native PAGE gels. Immunofluorescence microscopy revealed a flattened morphology of neutrophils associated with decondensed chromatin, remnants of granules in the cell periphery, and myeloperoxidase co-localized with extracellular DNA, features consistent with early-phase NETs. Nuclear staining revealed that a considerable fraction of bacterial cells in these structures were dead. The proteomes of two pathogens, Staphylococcus aureus and Escherichia coli, were indicative of adaptive responses to early-phase NETs, specifically the release of virulence factors and arrest of ribosomal protein synthesis. Finally, we discovered patterns of proteolysis consistent with widespread cleavage of proteins by neutrophil elastase, proteinase 3 and cathepsin G and evidence of citrullination in many nuclear proteins.

  5. Mechanism of interferon-gamma production by monocytes stimulated with myeloperoxidase and neutrophil extracellular traps.

    Science.gov (United States)

    Yamaguchi, Rui; Kawata, Jin; Yamamoto, Toshitaka; Ishimaru, Yasuji; Sakamoto, Arisa; Ono, Tomomichi; Narahara, Shinji; Sugiuchi, Hiroyuki; Hirose, Eiji; Yamaguchi, Yasuo

    2015-08-01

    Neutrophil extracellular traps (NETs) have an important role in antimicrobial innate immunity and release substances that may modulate the immune response. We investigated the effects of soluble factors from NETs and neutrophil granule proteins on human monocyte function by using the Transwell system to prevent cell-cell contact. NET formation was induced by exposing human neutrophils to phorbol myristate acetate (PMA). When monocytes were incubated with PMA alone, expression of interleukin (IL)-4, IL-6, IL-8, and tumor necrosis factor (TNF)-alpha mRNA was upregulated, but IL-10, IL-12, and interferon (IFN)-gamma mRNA were not detected. Incubation of monocytes with NETs enhanced the expression of IL-10 and IFN-gamma mRNA, but not IL-12 mRNA. Myeloperoxidase stimulated IFN-gamma production by monocytes in a dose-dependent manner. Both a nuclear factor-kappaB inhibitor (PDTC) and an intracellular calcium antagonist (TMB-8) prevented upregulation of IFN-gamma production. Neither a combined p38alpha and p38beta inhibitor (SB203580) nor an extracellular signal-regulated kinase inhibitor (PD98059) suppressed IFN-gamma production. Interestingly, a combined p38gamma and p38delta inhibitor (BIRB796) significantly decreased IFN-gamma production. These findings suggest that myeloperoxidase induces IFN-gamma production by monocytes via p38gamma/delta mitogen-activated protein kinase.

  6. Characterization of Early-Phase Neutrophil Extracellular Traps in Urinary Tract Infections

    Science.gov (United States)

    Yu, Yanbao; Kwon, Keehwan; Tsitrin, Tamara; Sikorski, Patricia; Nelson, Karen E.; Pieper, Rembert

    2017-01-01

    Neutrophils have an important role in the antimicrobial defense and resolution of urinary tract infections (UTIs). Our research suggests that a mechanism known as neutrophil extracellular trap (NET) formation is a defense strategy to combat pathogens that have invaded the urinary tract. A set of human urine specimens with very high neutrophil counts had microscopic evidence of cellular aggregation and lysis. Deoxyribonuclease I (DNase) treatment resulted in disaggregation of such structures, release of DNA fragments and a proteome enriched in histones and azurophilic granule effectors whose quantitative composition was similar to that of previously described in vitro-formed NETs. The effector proteins were further enriched in DNA-protein complexes isolated in native PAGE gels. Immunofluorescence microscopy revealed a flattened morphology of neutrophils associated with decondensed chromatin, remnants of granules in the cell periphery, and myeloperoxidase co-localized with extracellular DNA, features consistent with early-phase NETs. Nuclear staining revealed that a considerable fraction of bacterial cells in these structures were dead. The proteomes of two pathogens, Staphylococcus aureus and Escherichia coli, were indicative of adaptive responses to early-phase NETs, specifically the release of virulence factors and arrest of ribosomal protein synthesis. Finally, we discovered patterns of proteolysis consistent with widespread cleavage of proteins by neutrophil elastase, proteinase 3 and cathepsin G and evidence of citrullination in many nuclear proteins. PMID:28129394

  7. The selective estrogen receptor modulator raloxifene inhibits neutrophil extracellular trap formation.

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    Roxana Flores

    2016-12-01

    Full Text Available Raloxifene is a selective estrogen receptor modulator typically prescribed for the prevention/treatment of osteoporosis in postmenopausal women. Although raloxifene is known to have anti-inflammatory properties, its effect on human neutrophils, the primary phagocytic leukocytes of the immune system, remain poorly understood. Here, through a screen of pharmacologically active small molecules, we find that raloxifene prevents neutrophil cell death in response to the classical activator phorbol 12-myristate 13-acetate (PMA, a compound known to induce formation of DNA-based neutrophil extracellular traps (NETs. Inhibition of PMA-induced NET production by raloxifene was confirmed using quantitative and imaging-based assays. Human neutrophils from both male and female donors express the nuclear estrogen receptors ERα and ERβ, known targets of raloxifene. Like raloxifene, selective antagonists of these receptors inhibit PMA-induced NET production. Furthermore, raloxifene inhibited PMA-induced ERK phosphorylation but not reactive oxygen species (ROS production, pathways known to be key modulators of NET production. Finally, we found that raloxifene inhibited PMA-induced, NET-based killing of the leading human bacterial pathogen, methicillin-resistant Staphylococcus aureus (MRSA. Our results reveal that raloxifene is a potent modulator of neutrophil function and NET production.

  8. A role for muscarinic receptors in neutrophil extracellular trap formation and levamisole-induced autoimmunity

    Science.gov (United States)

    Carmona-Rivera, Carmelo; Purmalek, Monica M.; Moore, Erica; Waldman, Meryl; Walter, Peter J.; Garraffo, H. Martin; Phillips, Karran A.; Preston, Kenzie L.; Graf, Jonathan; Grayson, Peter C.

    2017-01-01

    Levamisole, an anthelmintic drug with cholinergic properties, has been implicated in cases of drug-induced vasculitis when added to cocaine for profit purposes. Neutrophil extracellular trap (NET) formation is a cell death mechanism characterized by extrusion of chromatin decorated with granule proteins. Aberrant NET formation and degradation have been implicated in idiopathic autoimmune diseases that share features with levamisole-induced autoimmunity as well as in drug-induced autoimmunity. This study’s objective was to determine how levamisole modulates neutrophil biology and its putative effects on the vasculature. Murine and human neutrophils exposed to levamisole demonstrated enhanced NET formation through engagement of muscarinic subtype 3 receptor. Levamisole-induced NETosis required activation of Akt and the RAF/MEK/ERK pathway, ROS induction through the nicotinamide adenine dinucleotide phosphate oxidase, and peptidylarginine deiminase activation. Sera from two cohorts of patients actively using levamisole-adulterated cocaine displayed autoantibodies against NET components. Cutaneous biopsy material obtained from individuals exposed to levamisole suggests that neutrophils produce NETs in areas of vasculitic inflammation and thrombosis. NETs generated by levamisole were toxic to endothelial cells and impaired endothelium-dependent vasorelaxation. Stimulation of muscarinic receptors on neutrophils by cholinergic agonists may contribute to the pathophysiology observed in drug-induced autoimmunity through the induction of inflammatory responses and neutrophil-induced vascular damage. PMID:28194438

  9. A role for muscarinic receptors in neutrophil extracellular trap formation and levamisole-induced autoimmunity.

    Science.gov (United States)

    Carmona-Rivera, Carmelo; Purmalek, Monica M; Moore, Erica; Waldman, Meryl; Walter, Peter J; Garraffo, H Martin; Phillips, Karran A; Preston, Kenzie L; Graf, Jonathan; Kaplan, Mariana J; Grayson, Peter C

    2017-02-09

    Levamisole, an anthelmintic drug with cholinergic properties, has been implicated in cases of drug-induced vasculitis when added to cocaine for profit purposes. Neutrophil extracellular trap (NET) formation is a cell death mechanism characterized by extrusion of chromatin decorated with granule proteins. Aberrant NET formation and degradation have been implicated in idiopathic autoimmune diseases that share features with levamisole-induced autoimmunity as well as in drug-induced autoimmunity. This study's objective was to determine how levamisole modulates neutrophil biology and its putative effects on the vasculature. Murine and human neutrophils exposed to levamisole demonstrated enhanced NET formation through engagement of muscarinic subtype 3 receptor. Levamisole-induced NETosis required activation of Akt and the RAF/MEK/ERK pathway, ROS induction through the nicotinamide adenine dinucleotide phosphate oxidase, and peptidylarginine deiminase activation. Sera from two cohorts of patients actively using levamisole-adulterated cocaine displayed autoantibodies against NET components. Cutaneous biopsy material obtained from individuals exposed to levamisole suggests that neutrophils produce NETs in areas of vasculitic inflammation and thrombosis. NETs generated by levamisole were toxic to endothelial cells and impaired endothelium-dependent vasorelaxation. Stimulation of muscarinic receptors on neutrophils by cholinergic agonists may contribute to the pathophysiology observed in drug-induced autoimmunity through the induction of inflammatory responses and neutrophil-induced vascular damage.

  10. β2 integrin mediates hantavirus-induced release of neutrophil extracellular traps.

    Science.gov (United States)

    Raftery, Martin J; Lalwani, Pritesh; Krautkrӓmer, Ellen; Peters, Thorsten; Scharffetter-Kochanek, Karin; Krüger, Renate; Hofmann, Jörg; Seeger, Karl; Krüger, Detlev H; Schönrich, Günther

    2014-06-30

    Rodent-borne hantaviruses are emerging human pathogens that cause severe human disease. The underlying mechanisms are not well understood, as hantaviruses replicate in endothelial and epithelial cells without causing any cytopathic effect. We demonstrate that hantaviruses strongly stimulated neutrophils to release neutrophil extracellular traps (NETs). Hantavirus infection induced high systemic levels of circulating NETs in patients and this systemic NET overflow was accompanied by production of autoantibodies to nuclear antigens. Analysis of the responsible mechanism using neutrophils from β2 null mice identified β2 integrin receptors as a master switch for NET induction. Further experiments suggested that β2 integrin receptors such as complement receptor 3 (CR3) and 4 (CR4) may act as novel hantavirus entry receptors. Using adenoviruses, we confirmed that viral interaction with β2 integrin induced strong NET formation. Collectively, β2 integrin-mediated systemic NET overflow is a novel viral mechanism of immunopathology that may be responsible for characteristic aspects of hantavirus-associated disease such as kidney and lung damage.

  11. Canine neutrophil extracellular traps release induced by the apicomplexan parasite Neospora Caninum in vitro

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    Zhengkai Wei

    2016-10-01

    Full Text Available Neosporosis is considered as one of the main causes of abortion and severe economic losses in dairy industry. The Canis genus serving as one of the confirmed definitive hosts of the apicomplexan parasite Neospora caninum (N. caninum plays a critical role in its life cycle. However, the effects of N. caninum on its definitive hosts of neutrophils extracellular traps (NETs formation remain unclear. In the present study, N. caninum tachyzoite-induced canine NETs formation was observed by scanning electron microscopy (SEM. Visualization of DNA decorated with H3, NE and MPO within N. caninum tachyzoite-induced NETs were examined using fluorescence confocal microscopy analyses. Furthermore, the formation of canine NETs was quantified using Sytox Green staining, and the LDH levels in supernatants were examined by an LDH Cytotoxicity Assay® kit. The results clearly showed that NETs-like structures were induced by N. caninum tachyzoites, and the major components within these structures induced by N. caninum tachyzoite were further confirmed by fluorescence confocal microscopy visualization. These results suggest that N. caninum tachyzoites strongly induced NETs formation in canine PMN. In functional inhibition assays, the blockings of NADPH oxidase, NE, MPO, SOCE, ERK 1/2 and p38 MAPK signaling pathways significantly inhibited N. caninum tachyzoite-induced NETs formation, which suggests that N. caninum tachyzoite-induced NETs formation is a NADPH oxidase-, NE-, MPO-, SOCE-, ERK 1/2- and p38 MAPK-dependent cell death process. To our knowledge, this study is the first to report the formation of NETs in canine PMN against N. caninum infection.

  12. Identification of macrophage extracellular trap-like structures in mammary gland adipose tissue: a preliminary study.

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    Sunish eMohanan

    2013-03-01

    Full Text Available PAD4-mediated hypercitrullination of histone H4 arginine 3 (H4R3 has been previously found to promote the formation of Neutrophil Extracellular Traps (NET in inflamed tissues and the resulting histone H4 citrulline 3 (H4Cit3 modification is thought to play a key role in extracellular trap (ET formation by promoting chromatin decondensation. In addition to neutrophils, macrophages have also recently been found to generate functional extracellular traps (METs. However, a role for PADs in ET formation in macrophages has not been previously described. Transcripts for PAD2 and PAD4 are found in mature macrophages and these cells can be induced to citrullinate proteins, thus raising the possibility that PADs may play a direct role in ET formation in macrophages via histone hypercitrullination. In breast and visceral white adipose tissue from obese patients, infiltrating macrophages are often seen to surround dead adipocytes forming characteristic crown-like structures (CLS and the presence of these lesions is associated with increased levels of inflammatory mediators. In light of these observations, we have initiated studies to test whether PADs are expressed in CLS macrophages and whether these macrophages might form METs. Our preliminary findings show that PAD2 (and to a lesser extent, PAD4 is expressed in both in the macrophage cell line (RAW 264.7 and in CLS lesions. Additionally, we provide evidence that macrophage-derived extracellular histones are seen around presumptive macrophages within CLS lesions and that these histones contain the H4Cit3 modification. These initial findings support our hypothesis that obesity-induced adipose tissue inflammation promotes the formation of METs within CLS lesions via PAD-mediated histone hypercitrullination. Subsequent studies are underway to further validate these findings and to investigate the role in PAD-mediated MET formation in CLS function in the mammary gland.

  13. Induction of Neutrophil Extracellular Traps in Shiga Toxin-Associated Hemolytic Uremic Syndrome.

    Science.gov (United States)

    Ramos, Maria Victoria; Mejias, Maria Pilar; Sabbione, Florencia; Fernandez-Brando, Romina Jimena; Santiago, Adriana Patricia; Amaral, Maria Marta; Exeni, Ramon; Trevani, Analia Silvina; Palermo, Marina Sandra

    2016-01-01

    Hemolytic uremic syndrome (HUS), a vascular disease characterized by hemolytic anemia, thrombocytopenia, and acute renal failure, is caused by enterohemorrhagic Shiga toxin (Stx)-producing bacteria, which mainly affect children. Besides Stx, the inflammatory response mediated by neutrophils (PMN) is essential to HUS evolution. PMN can release neutrophil extracellular traps (NET) composed of DNA, histones, and other proteins. Since NET are involved in infectious and inflammatory diseases, the aim of this work was to investigate the contribution of NET to HUS. Plasma from HUS patients contained increased levels of circulating free-DNA and nucleosomes in comparison to plasma from healthy children. Neutrophils from HUS patients exhibited a greater capacity to undergo spontaneous NETosis. NET activated human glomerular endothelial cells, stimulating secretion of the proinflammatory cytokines IL-6 and IL-8. Stx induced PMN activation as judged by its ability to trigger reactive oxygen species production, increase CD11b and CD66b expression, and induce NETosis in PMN from healthy donors. During HUS, NET can contribute to the inflammatory response and thrombosis in the microvasculature and thus to renal failure. Intervention strategies to inhibit inflammatory mechanisms mediated by PMN, such as NETosis, could have a potential therapeutic impact towards amelioration of the severity of HUS.

  14. Neutrophil extracellular traps downregulate lipopolysaccharide-induced activation of monocyte-derived dendritic cells.

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    Barrientos, Lorena; Bignon, Alexandre; Gueguen, Claire; de Chaisemartin, Luc; Gorges, Roseline; Sandré, Catherine; Mascarell, Laurent; Balabanian, Karl; Kerdine-Römer, Saadia; Pallardy, Marc; Marin-Esteban, Viviana; Chollet-Martin, Sylvie

    2014-12-01

    Polymorphonuclear neutrophils (PMN) play a central role in inflammation and participate in its control, notably by modulating dendritic cell (DC) functions via soluble mediators or cell-cell contacts. Neutrophil extracellular traps (NETs) released by PMN could play a role in this context. To evaluate NET effects on DC maturation, we developed a model based on monocyte-derived DC (moDC) and calibrated NETs isolated from fresh human PMN. We found that isolated NETs alone had no discernable effect on moDC. In contrast, they downregulated LPS-induced moDC maturation, as shown by decreased surface expression of HLA-DR, CD80, CD83, and CD86, and by downregulated cytokine production (TNF-α, IL-6, IL-12, IL-23), with no increase in the expression of tolerogenic DC genes. Moreover, the presence of NETs during moDC maturation diminished the capacity of these moDC to induce T lymphocyte proliferation in both autologous and allogeneic conditions, and modulated CD4(+) T lymphocyte polarization by promoting the production of Th2 cytokines (IL-5 and IL-13) and reducing that of Th1 and Th17 cytokines (IFN-γ and IL-17). Interestingly, the expression and activities of the lymphoid chemokine receptors CCR7 and CXCR4 on moDC were not altered when moDC matured in the presence of NETs. Together, these findings reveal a new role for NETs in adaptive immune responses, modulating some moDC functions and thereby participating in the control of inflammation.

  15. Autophagy is induced by anti-neutrophil cytoplasmic Abs and promotes neutrophil extracellular traps formation.

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    Sha, Li-Li; Wang, Huan; Wang, Chen; Peng, Hong-Ying; Chen, Min; Zhao, Ming-Hui

    2016-11-01

    Dysregulated neutrophil extracellular traps (NETs) formation contributes to the pathogenesis of anti-neutrophil cytoplasmic Ab (ANCA)-associated vasculitis (AAV). Increasing evidence indicates that autophagy is involved in the process of NETs formation. In this study, we aimed to investigate whether ANCA could induce autophagy in the process of NETs formation. Autophagy was detected using live cell imaging, microtubule-associated protein light chain 3B (LC3B) accumulation and Western blotting. The results showed that autophagy vacuolization was detected in neutrophils treated with ANCA-positive IgG by live cell imaging. This effect was enhanced by rapamycin, the autophagy inducer, and weakened by 3-methyladenine (3-MA), the autophagy inhibitor. In line with these results, the autophagy marker, LC3B, showed a punctate distribution pattern in the neutrophils stimulated with ANCA-positive IgG. In the presence of rapamycin, LC3B accumulation was further increased; however, this effect was attenuated by 3-MA. Moreover, incubated with ANCA-positive IgG, the NETosis rate significantly increased compared with the unstimulated group. And, the rate significantly increased or decreased in the neutrophils pretreated with rapamycin or 3-MA, respectively, as compared with the cells incubated with ANCA-positive IgG. Overall, this study demonstrates that autophagy is induced by ANCA and promotes ANCA-induced NETs formation.

  16. Efficient neutrophil extracellular trap induction requires mobilization of both intracellular and extracellular calcium pools and is modulated by cyclosporine A.

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    Anurag Kumar Gupta

    Full Text Available Excessive or aberrant generation of neutrophil extracellular traps (NETs has recently become implicated in the underlying aetiology of a number of human pathologies including preeclampsia, systemic lupus erythromatosus, rheumatoid arthritis, auto-antibody induced small vessel vasculitis, coagulopathies such as deep vein thrombosis or pulmonary complications. These results imply that effective pharmacological therapeutic strategies will need to be developed to counter overt NETosis in these and other inflammatory disorders. As calcium flux is implicated in the generation of reactive oxygen species and histone citrullination, two key events in NETosis, we analysed the roles of both extra- and intracellular calcium pools and their modulation by pharmacological agents in the NETotic process in detail. Interleukin-8 (IL-8 was used as a physiological stimulus of NETosis. Our data demonstrate that efficient induction of NETosis requires mobilisation of both extracellular and intracellular calcium pools. Since modulation of the calcineurin pathway by cyclosporine A has been described in neutrophils, we investigated its influence on NETosis. Our data indicate that IL-8 induced NETosis is reduced by ascomycin and cyclosporine A, antagonists of the calcineurin pathway, but not following treatment with rapamycin, which utilizes the mTOR pathway. The action of the G protein coupled receptor phospholipase C pathway appears to be essential for the induction of NETs by IL-8, as NETosis was diminished by treatment with either pertussis toxin, a G-protein inhibitor, the phospholipase C inhibitor, U73122, or staurosporine, an inhibitor of protein kinase C. The data regarding the calcineurin antagonists, ascomycin and cyclosporine A, open the possibility to therapeutically suppress or modulate NETosis. They also provide new insight into the mechanism whereby such immune suppressive drugs render transplant patients susceptible to opportunistic fungal infections.

  17. Iron-chelating agent, deferasirox, inhibits neutrophil activation and extracellular trap formation.

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    Kono, Mari; Saigo, Katsuyasu; Yamamoto, Shiori; Shirai, Kohei; Iwamoto, Shuta; Uematsu, Tomoko; Takahashi, Takayuki; Imoto, Shion; Hashimoto, Makoto; Minami, Yosuke; Wada, Atsushi; Takenokuchi, Mariko; Kawano, Seiji

    2016-10-01

    Iron-chelating agents, which are frequently prescribed to transfusion-dependent patients, have various useful biological effects in addition to chelation. Reactive oxygen species (ROS) produced by neutrophils can cause pulmonary endothelial cell damage, which can lead to acute lung injury (ALI). We previously reported that deferasirox (DFS), an iron-chelating agent, inhibits phorbol myristate acetate (PMA) or formyl-methionyl-leucyl-phenylalanine (fMLP)-induced ROS production in neutrophils, in vitro. Here, we investigate whether DFS inhibits vacuolization in neutrophils and neutrophil extracellular trap (NET) formation. Human neutrophils were incubated with DFS and stimulated with PMA or fMLP. Human neutrophils were separated from heparinized peripheral blood using density gradient centrifugation, and subsequently incubated with DFS. After 10 minutes, neutrophils were stimulated by PMA or fMLP. Vacuole formation was observed by electron microscopy. For observing NET formations using microscopes, immunohistological analyses using citrullinated histone H3 and myeloperoxidase antibodies, and SYTOX Green (an impermeable DNA detection dye) staining, were conducted. NET formation was measured as the quantity of double-stranded DNA (dsDNA), using the AccuBlue Broad Range dsDNA Quantitation Kit. DFS (50 μmol/L) inhibited vacuole formation in the cytoplasm and NET formation. Additionally, 5-100 μmol/L concentration of DFS inhibited the release of dsDNA in a dose-independent manner. We demonstrate that DFS inhibits not only ROS production but also vacuolization and NET formation in neutrophils. These results suggest the possibility of protective effects of DFS against NET-related adverse effects, including ALI and thrombosis.

  18. Low-Frequency Electromagnetic Field Exposure Enhances Extracellular Trap Formation by Human Neutrophils through the NADPH Pathway.

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    Golbach, Lieke A; Scheer, Marleen H; Cuppen, Jan J M; Savelkoul, Huub; Verburg-van Kemenade, B M Lidy

    2015-01-01

    Low-frequency (LF) electromagnetic fields (EMFs) are abundantly present in modern society, and the potential biological consequences of exposure to these fields are under intense debate. Immune cells are suggested as possible target cells, though a clear mechanism is lacking. Considering their crucial role in innate immune activation, we selected an ex vivo exposure set-up with human neutrophils to investigate a possible correlation between neutrophil extracellular trap (NET) formation and LF EMF exposure. Our study shows that formation of NETs is enhanced by LF EMF exposure. Enhanced NET formation leads to increased antimicrobial properties as well as damage to surrounding cells. We found that LF-EMF-induced NET formation is dependent on the NADPH oxidase pathway and production of reactive oxygen species. Additionally, LF EMF exposure does not influence autophagy and PAD4 activity. Our study provides a mechanism by which exposure to LF EMFs could influence the innate immune system.

  19. The Role of Reactive Oxygen Species (ROS in the Formation of Extracellular Traps (ETs in Humans

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    Walter Stoiber

    2015-05-01

    Full Text Available Extracellular traps (ETs are reticulate structures of extracellular DNA associated with antimicrobial molecules. Their formation by phagocytes (mainly by neutrophils: NETs has been identified as an essential element of vertebrate innate immune defense. However, as ETs are also toxic to host cells and potent triggers of autoimmunity, their role between pathogen defense and human pathogenesis is ambiguous, and they contribute to a variety of acute and chronic inflammatory diseases. Since the discovery of ET formation (ETosis a decade ago, evidence has accumulated that most reaction cascades leading to ET release involve ROS. An important new facet was added when it became apparent that ETosis might be directly linked to, or be a variant of, the autophagy cell death pathway. The present review analyzes the evidence to date on the interplay between ROS, autophagy and ETosis, and highlights and discusses several further aspects of the ROS-ET relationship that are incompletely understood. These aspects include the role of NADPH oxidase-derived ROS, the molecular requirements of NADPH oxidase-dependent ETosis, the roles of NADPH oxidase subtypes, extracellular ROS and of ROS from sources other than NADPH oxidase, and the present evidence for ROS-independent ETosis. We conclude that ROS interact with ETosis in a multidimensional manner, with influence on whether ETosis shows beneficial or detrimental effects.

  20. Placental histology and neutrophil extracellular traps in lupus and pre-eclampsia pregnancies

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    Marder, Wendy; Knight, Jason S; Kaplan, Mariana J; Somers, Emily C; Zhang, Xu; O'Dell, Alexander A; Padmanabhan, Vasantha; Lieberman, Richard W

    2016-01-01

    Objective Systemic lupus erythematosus (SLE) is associated with increased risk of adverse pregnancy outcomes, including pre-eclampsia, particularly in association with antiphospholipid antibody syndrome (APS). While significant placental abnormalities are expected in pre-eclampsia, less is known about how lupus activity and APS in pregnancy affect the placenta. We describe placental pathology from a population of lupus pregnancies, several of which were complicated by APS-related thromboses, in which pre-eclampsia and other complications developed. We performed standard histopathological placental review and quantified neutrophils and neutrophil extracellular traps (NETs) in the intervillous space, given the recognised association of NETs with lupus, APS and pre-eclampsia. Methods Pre-eclampsia, SLE and control placentas were scored for histological features, and neutrophils were quantified on H&E and immunohistochemical staining for the granular protein myeloperoxidase. NETs were identified by extracellular myeloperoxidase staining in the setting of decondensed nuclei. Non-parametric analysis was used to evaluate differences in netting and intact neutrophils between groups, with Kruskal–Wallis testing for associations between histological findings and neutrophils. Results Placentas were evaluated from 35 pregnancies: 10 controls, 11 pre-eclampsia, 4 SLE+pre-eclampsia and 10 SLE, including one complicated by catastrophic APS and one complicated by hepatic and splenic vein thromboses during pregnancy. Intrauterine growth restriction and oligohydramnios were observed in lupus cases but not controls. Significantly more NETs were found infiltrating placental intervillous spaces in pre-eclampsia, SLE+pre-eclampsia and all 10 SLE non-pre-eclampsia cases. The ratio of NETs to total neutrophils was significantly increased in all case groups compared with controls. When present, NETs were associated with maternal vasculitis, laminar decidual necrosis, maternal

  1. The Fungal Exopolysaccharide Galactosaminogalactan Mediates Virulence by Enhancing Resistance to Neutrophil Extracellular Traps.

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    Mark J Lee

    2015-10-01

    Full Text Available Of the over 250 Aspergillus species, Aspergillus fumigatus accounts for up to 80% of invasive human infections. A. fumigatus produces galactosaminogalactan (GAG, an exopolysaccharide composed of galactose and N-acetyl-galactosamine (GalNAc that mediates adherence and is required for full virulence. Less pathogenic Aspergillus species were found to produce GAG with a lower GalNAc content than A. fumigatus and expressed minimal amounts of cell wall-bound GAG. Increasing the GalNAc content of GAG of the minimally pathogenic A. nidulans, either through overexpression of the A. nidulans epimerase UgeB or by heterologous expression of the A. fumigatus epimerase Uge3 increased the amount of cell wall bound GAG, augmented adherence in vitro and enhanced virulence in corticosteroid-treated mice to levels similar to A. fumigatus. The enhanced virulence of the overexpression strain of A. nidulans was associated with increased resistance to NADPH oxidase-dependent neutrophil extracellular traps (NETs in vitro, and was not observed in neutropenic mice or mice deficient in NADPH-oxidase that are unable to form NETs. Collectively, these data suggest that cell wall-bound GAG enhances virulence through mediating resistance to NETs.

  2. New aspects on the structure of neutrophil extracellular traps from chronic obstructive pulmonary disease and in vitro generation.

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    Astrid Obermayer

    Full Text Available Polymorphonuclear neutrophils have in recent years attracted new attention due to their ability to release neutrophil extracellular traps (NETs. These web-like extracellular structures deriving from nuclear chromatin have been depicted in ambiguous roles between antimicrobial defence and host tissue damage. NETs consist of DNA strands of varying thickness and are decorated with microbicidal and cytotoxic proteins. Their principal structure has in recent years been characterised at molecular and ultrastructural levels but many features that are of direct relevance to cytotoxicity are still incompletely understood. These include the extent of chromatin decondensation during NET formation and the relative amounts and spatial distribution of the microbicidal components within the NET. In the present work, we analyse the structure of NETs found in induced sputum of patients with acutely exacerbated chronic obstructive pulmonary disease (COPD using confocal laser microscopy and electron microscopy. In vitro induced NETs from human neutrophils serve for purposes of comparison and extended analysis of NET structure. Results demonstrate that COPD sputa are characterised by the pronounced presence of NETs and NETotic neutrophils. We provide new evidence that chromatin decondensation during NETosis is most extensive and generates substantial amounts of double-helix DNA in 'beads-on-a-string' conformation. New information is also presented on the abundance and location of neutrophil elastase (NE and citrullinated histone H3 (citH3. NE occurs in high densities in nearly all non-fibrous constituents of the NETs while citH3 is much less abundant. We conclude from the results that (i NETosis is an integral part of COPD pathology; this is relevant to all future research on the etiology and therapy of the disease; and that (ii release of 'beads-on-a-string' DNA studded with non-citrullinated histones is a common feature of in vivo NETosis; this is of relevance to

  3. The architecture of neutrophil extracellular traps investigated by atomic force microscopy

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    Pires, Ricardo H.; Felix, Stephan B.; Delcea, Mihaela

    2016-07-01

    Neutrophils are immune cells that engage in a suicidal pathway leading to the release of partially decondensed chromatin, or neutrophil extracellular traps (NETs). NETs behave as a double edged sword; they can bind to pathogens thereby ensnaring them and limiting their spread during infection; however, they may bind to host circulating materials and trigger thrombotic events, and are associated with autoimmune disorders. Despite the fundamental role of NETs as part of an immune system response, there is currently a very poor understanding of how their nanoscale properties are reflected in their macroscopic impact. In this work, using a combination of fluorescence and atomic force microscopy, we show that NETs appear as a branching filament network that results in a substantially organized porous structure with openings with 0.03 +/- 0.04 μm2 on average and thus in the size range of small pathogens. Topological profiles typically up to 3 +/- 1 nm in height are compatible with a ``beads on a string'' model of nucleosome chromatin. Typical branch lengths of 153 +/- 103 nm appearing as rigid rods and height profiles of naked DNA in NETs of 1.2 +/- 0.5 nm are indicative of extensive DNA supercoiling throughout NETs. The presence of DNA duplexes could also be inferred from force spectroscopy and the occurrence of force plateaus that ranged from ~65 pN to 300 pN. Proteolytic digestion of NETs resulted in widespread disassembly of the network structure and considerable loss of mechanical properties. Our results suggest that the underlying structure of NETs is considerably organized and that part of its protein content plays an important role in maintaining its mesh architecture. We anticipate that NETs may work as microscopic mechanical sieves with elastic properties that stem from their DNA-protein composition, which is able to segregate particles also as a result of their size. Such a behavior may explain their participation in capturing pathogens and their association

  4. 中性粒细胞网的研究进展%Research progress on neutrophil extracellular traps

    Institute of Scientific and Technical Information of China (English)

    赵赛男; 李文卿

    2011-01-01

    中性粒细胞通过吞噬、脱颗粒和形成中性粒细胞网(NET)等方式抵抗微生物感染。自NET被发现后,研究者们即从一个全新的视角来诠释固有免疫控制和杀伤微生物的机制。本文就NET的结构、形成和在疾病中的作用以及微生物逃逸NET的作用作一综述。%It is believed that neutrophils have three major strategies to resist microbian infection, including phagocytosis, degranulation, and neutrophil extracellular traps (NET) formation. The discovery of NET introduces a brand-new perspective for the researchers from which they make analysis of the mechanism of innate immune system trapping and killing microorganisms. The thesis aims to give a review of NET's structure and formation, as well as the faction in disease and the escape of microorganisms from NET.

  5. Neutrophil Attack Triggers Extracellular Trap-Dependent Candida Cell Wall Remodeling and Altered Immune Recognition.

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    Alex Hopke

    2016-05-01

    Full Text Available Pathogens hide immunogenic epitopes from the host to evade immunity, persist and cause infection. The opportunistic human fungal pathogen Candida albicans, which can cause fatal disease in immunocompromised patient populations, offers a good example as it masks the inflammatory epitope β-glucan in its cell wall from host recognition. It has been demonstrated previously that β-glucan becomes exposed during infection in vivo but the mechanism behind this exposure was unknown. Here, we show that this unmasking involves neutrophil extracellular trap (NET mediated attack, which triggers changes in fungal cell wall architecture that enhance immune recognition by the Dectin-1 β-glucan receptor in vitro. Furthermore, using a mouse model of disseminated candidiasis, we demonstrate the requirement for neutrophils in triggering these fungal cell wall changes in vivo. Importantly, we found that fungal epitope unmasking requires an active fungal response in addition to the stimulus provided by neutrophil attack. NET-mediated damage initiates fungal MAP kinase-driven responses, particularly by Hog1, that dynamically relocalize cell wall remodeling machinery including Chs3, Phr1 and Sur7. Neutrophil-initiated cell wall disruptions augment some macrophage cytokine responses to attacked fungi. This work provides insight into host-pathogen interactions during disseminated candidiasis, including valuable information about how the C. albicans cell wall responds to the biotic stress of immune attack. Our results highlight the important but underappreciated concept that pattern recognition during infection is dynamic and depends on the host-pathogen dialog.

  6. Human Leukocytes Kill Brugia malayi Microfilariae Independently of DNA-Based Extracellular Trap Release.

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    Ciaran J McCoy

    2017-01-01

    Full Text Available Wuchereria bancrofti, Brugia malayi and Brugia timori infect over 100 million people worldwide and are the causative agents of lymphatic filariasis. Some parasite carriers are amicrofilaremic whilst others facilitate mosquito-based disease transmission through blood-circulating microfilariae (Mf. Recent findings, obtained largely from animal model systems, suggest that polymorphonuclear leukocytes (PMNs contribute to parasitic nematode-directed type 2 immune responses. When exposed to certain pathogens PMNs release extracellular traps (NETs in the form of chromatin loaded with various antimicrobial molecules and proteases.In vitro, PMNs expel large amounts of NETs that capture but do not kill B. malayi Mf. NET morphology was confirmed by fluorescence imaging of worm-NET aggregates labelled with DAPI and antibodies to human neutrophil elastase, myeloperoxidase and citrullinated histone H4. A fluorescent, extracellular DNA release assay was used to quantify and observe Mf induced NETosis over time. Blinded video analyses of PMN-to-worm attachment and worm survival during Mf-leukocyte co-culture demonstrated that DNase treatment eliminates PMN attachment in the absence of serum, autologous serum bolsters both PMN attachment and PMN plus peripheral blood mononuclear cell (PBMC mediated Mf killing, and serum heat inactivation inhibits both PMN attachment and Mf killing. Despite the effects of heat inactivation, the complement inhibitor compstatin did not impede Mf killing and had little effect on PMN attachment. Both human PMNs and monocytes, but not lymphocytes, are able to kill B. malayi Mf in vitro and NETosis does not significantly contribute to this killing. Leukocytes derived from presumably parasite-naïve U.S. resident donors vary in their ability to kill Mf in vitro, which may reflect the pathological heterogeneity associated with filarial parasitic infections.Human innate immune cells are able to recognize, attach to and kill B. malayi

  7. Human Leukocytes Kill Brugia malayi Microfilariae Independently of DNA-Based Extracellular Trap Release

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    McCoy, Ciaran J.; Reaves, Barbara J.; Giguère, Steeve; Coates, Ruby; Rada, Balázs

    2017-01-01

    Background Wuchereria bancrofti, Brugia malayi and Brugia timori infect over 100 million people worldwide and are the causative agents of lymphatic filariasis. Some parasite carriers are amicrofilaremic whilst others facilitate mosquito-based disease transmission through blood-circulating microfilariae (Mf). Recent findings, obtained largely from animal model systems, suggest that polymorphonuclear leukocytes (PMNs) contribute to parasitic nematode-directed type 2 immune responses. When exposed to certain pathogens PMNs release extracellular traps (NETs) in the form of chromatin loaded with various antimicrobial molecules and proteases. Principal findings In vitro, PMNs expel large amounts of NETs that capture but do not kill B. malayi Mf. NET morphology was confirmed by fluorescence imaging of worm-NET aggregates labelled with DAPI and antibodies to human neutrophil elastase, myeloperoxidase and citrullinated histone H4. A fluorescent, extracellular DNA release assay was used to quantify and observe Mf induced NETosis over time. Blinded video analyses of PMN-to-worm attachment and worm survival during Mf-leukocyte co-culture demonstrated that DNase treatment eliminates PMN attachment in the absence of serum, autologous serum bolsters both PMN attachment and PMN plus peripheral blood mononuclear cell (PBMC) mediated Mf killing, and serum heat inactivation inhibits both PMN attachment and Mf killing. Despite the effects of heat inactivation, the complement inhibitor compstatin did not impede Mf killing and had little effect on PMN attachment. Both human PMNs and monocytes, but not lymphocytes, are able to kill B. malayi Mf in vitro and NETosis does not significantly contribute to this killing. Leukocytes derived from presumably parasite-naïve U.S. resident donors vary in their ability to kill Mf in vitro, which may reflect the pathological heterogeneity associated with filarial parasitic infections. Conclusions/Significance Human innate immune cells are able to

  8. Comparison of Bottomless Lift Nets and Breder Traps for Sampling Salt-Marsh Nekton

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    U.S. Environmental Protection Agency — Data set contains: the length of mummichogs (Fundulus heteroclitus) caught on lift nets and Breder traps from May to September 2002; the sizes of green crabs caught...

  9. Report on a Cannon Net Trap Workshop Held at Swan Lake National Wildlife Refuge

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    US Fish and Wildlife Service, Department of the Interior — The following is a report of material covered during a workshop on cannon net trapping and banding held at the Swan Lake National Wildlife Refuge, Sumner, Missouri,...

  10. Phenol-Soluble Modulin α Peptide Toxins from Aggressive Staphylococcus aureus Induce Rapid Formation of Neutrophil Extracellular Traps through a Reactive Oxygen Species-Independent Pathway

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    Björnsdottir, Halla; Dahlstrand Rudin, Agnes; Klose, Felix P.; Elmwall, Jonas; Welin, Amanda; Stylianou, Marios; Christenson, Karin; Urban, Constantin F.; Forsman, Huamei; Dahlgren, Claes; Karlsson, Anna; Bylund, Johan

    2017-01-01

    Neutrophils have the ability to capture and kill microbes extracellularly through the formation of neutrophil extracellular traps (NETs). These are DNA and protein structures that neutrophils release extracellularly and are believed to function as a defense mechanism against microbes. The classic NET formation process, triggered by, e.g., bacteria, fungi, or by direct stimulation of protein kinase C through phorbol myristate acetate, is an active process that takes several hours and relies on the production of reactive oxygen species (ROS) that are further modified by myeloperoxidase (MPO). We show here that NET-like structures can also be formed by neutrophils after interaction with phenol-soluble modulin α (PSMα) that are cytotoxic membrane-disturbing peptides, secreted from community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA). The PSMα-induced NETs contained the typical protein markers and were able to capture microbes. The PSMα-induced NET structures were disintegrated upon prolonged exposure to DNase-positive S. aureus but not on exposure to DNase-negative Candida albicans. Opposed to classic NETosis, PSMα-triggered NET formation occurred very rapidly, independently of ROS or MPO, and was also manifest at 4°C. These data indicate that rapid NETs release may result from cytotoxic membrane disturbance by PSMα peptides, a process that may be of importance for CA-MRSA virulence. PMID:28337204

  11. Neutrophil extracellular trap formation is associated with IL-1β and autophagy-related signaling in gout.

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    Ioannis Mitroulis

    Full Text Available BACKGROUND: Gout is a prevalent inflammatory arthritis affecting 1-2% of adults characterized by activation of innate immune cells by monosodium urate (MSU crystals resulting in the secretion of interleukin-1β (IL-1β. Since neutrophils play a major role in gout we sought to determine whether their activation may involve the formation of proinflammatory neutrophil extracellular traps (NETs in relation to autophagy and IL-1β. METHODOLOGY/PRINCIPAL FINDINGS: Synovial fluid neutrophils from six patients with gout crisis and peripheral blood neutrophils from six patients with acute gout and six control subjects were isolated. MSU crystals, as well as synovial fluid or serum obtained from patients with acute gout, were used for the treatment of control neutrophils. NET formation was assessed using immunofluorescence microscopy. MSU crystals or synovial fluid or serum from patients induced NET formation in control neutrophils. Importantly, NET production was observed in neutrophils isolated from synovial fluid or peripheral blood from patients with acute gout. NETs contained the alarmin high mobility group box 1 (HMGB1 supporting their pro-inflammatory potential. Inhibition of phosphatidylinositol 3-kinase signaling or phagolysosomal fusion prevented NET formation, implicating autophagy in this process. NET formation was driven at least in part by IL-1β as demonstrated by experiments involving IL-1β and its inhibitor anakinra. CONCLUSIONS/SIGNIFICANCE: These findings document for the first time that activation of neutrophils in gout is associated with the formation of proinflammatory NETs and links this process to both autophagy and IL-1β. Modulation of the autophagic machinery may represent an additional therapeutic study in crystalline arthritides.

  12. Effects of Aggregatibacter actinomycetemcomitans leukotoxin on neutrophil migration and extracellular trap formation

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    Josefine Hirschfeld

    2016-11-01

    Full Text Available Background: Aggressive periodontitis is associated with the presence of Aggregatibacter actinomycetemcomitans, a leukotoxin (Ltx-producing periodontal pathogen. Ltx has the ability to lyse white blood cells including neutrophils. Objectives: This study was aimed at investigating the interactions between neutrophils and Ltx with regard to the chemotactic properties of Ltx and the release of neutrophil extracellular traps (NETs. Methods: Neutrophils from healthy blood donors were isolated and incubated for 30 min and 3 h with increasing concentrations of Ltx (1, 10, and 100 ng/mL as well as with A. actinomycetemcomitans strains (NCTC 9710 and HK 1651 producing different levels of Ltx. Formation of NETs and cell lysis were assessed by microscopy, fluorescence-based assays, and measurement of released lactate dehydrogenase. Neutrophil migration in response to different Ltx gradients was monitored by real-time video microscopy, and image analysis was performed using ImageJ software. Results: Although Ltx (10 and 100 ng/mL and the leukotoxic A. actinomycetemcomitans strain HK 1651 lysed some neutrophils, other cells were still capable of performing NETosis in a concentration-dependent manner. Low doses of Ltx and the weakly leukotoxic strain NCTC 9710 did not lead to neutrophil lysis, but did induce some NETosis. Furthermore, all three concentrations of Ltx enhanced random neutrophil movement; however, low directional accuracy was observed compared with the positive control (fMLP. Conclusions: The results indicate that Ltx acts both as a neutrophil activator and also causes cell death. In addition, Ltx directly induces NETosis in neutrophils prior to cell lysis. In future studies, the underlying pathways involved in Ltx-meditated neutrophil activation and NETosis need to be investigated further.

  13. Effects of Aggregatibacter actinomycetemcomitans leukotoxin on neutrophil migration and extracellular trap formation

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    Hirschfeld, Josefine; Roberts, Helen M.; Chapple, Iain L. C.; Parčina, Marijo; Jepsen, Søren; Johansson, Anders; Claesson, Rolf

    2016-01-01

    Background Aggressive periodontitis is associated with the presence of Aggregatibacter actinomycetemcomitans, a leukotoxin (Ltx)-producing periodontal pathogen. Ltx has the ability to lyse white blood cells including neutrophils. Objectives This study was aimed at investigating the interactions between neutrophils and Ltx with regard to the chemotactic properties of Ltx and the release of neutrophil extracellular traps (NETs). Methods Neutrophils from healthy blood donors were isolated and incubated for 30 min and 3 h with increasing concentrations of Ltx (1, 10, and 100 ng/mL) as well as with A. actinomycetemcomitans strains (NCTC 9710 and HK 1651) producing different levels of Ltx. Formation of NETs and cell lysis were assessed by microscopy, fluorescence-based assays, and measurement of released lactate dehydrogenase. Neutrophil migration in response to different Ltx gradients was monitored by real-time video microscopy, and image analysis was performed using ImageJ software. Results Although Ltx (10 and 100 ng/mL) and the leukotoxic A. actinomycetemcomitans strain HK 1651 lysed some neutrophils, other cells were still capable of performing NETosis in a concentration-dependent manner. Low doses of Ltx and the weakly leukotoxic strain NCTC 9710 did not lead to neutrophil lysis, but did induce some NETosis. Furthermore, all three concentrations of Ltx enhanced random neutrophil movement; however, low directional accuracy was observed compared with the positive control (fMLP). Conclusions The results indicate that Ltx acts both as a neutrophil activator and also causes cell death. In addition, Ltx directly induces NETosis in neutrophils prior to cell lysis. In future studies, the underlying pathways involved in Ltx-meditated neutrophil activation and NETosis need to be investigated further. PMID:27834173

  14. Integrin-dependent cell adhesion to neutrophil extracellular traps through engagement of fibronectin in neutrophil-like cells

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    Monti, Marcello; Iommelli, Francesca; De Rosa, Viviana; Carriero, Maria Vincenza; Miceli, Roberta; Camerlingo, Rosa; Di Minno, Giovanni; Del Vecchio, Silvana

    2017-01-01

    Neutrophil extracellular traps (NETs), originally recognized as a host defense mechanism, were reported to promote thrombosis and metastatic dissemination of cancer cells. Here we tested the role of integrins α5β1 and ανβ3 in the adhesion of cancer cells to NETs. Neutrophil-like cells stimulated with calcium ionophore (A23187) were used as a stable source of cell-free NETs-enriched suspensions. Using NETs as an adhesion substrate, two human K562 cell lines, differentially expressing α5β1 and ανβ3 integrins, were subjected to adhesion assays in the presence or absence of DNAse 1, blocking antibodies against α5β1 or ανβ3, alone or in combination with DNAse 1, and Proteinase K. As expected DNAse 1 treatment strongly inhibited adhesion of both cell lines to NETs. An equivalent significant reduction of cell adhesion to NETs was obtained after treatment of cells with blocking antibodies against α5β1 or ανβ3 indicating that both integrins were able to mediate cell adhesion to NETs. Furthermore, the combination of DNAse 1 and anti-integrin antibody treatment almost completely blocked cell adhesion. Western blot analysis and immunoprecipitation experiments showed a dose-dependent increase of fibronectin levels in samples from stimulated neutrophil-like cells and a direct or indirect interaction of fibronectin with histone H3. Finally, co-immunolocalization studies with confocal microscopy showed that fibronectin and citrullinated histone H3 co-localize inside the web-structure of NETs. In conclusion, our study showed that α5β1 and ανβ3 integrins mediate cell adhesion to NETs by binding to their common substrate fibronectin. Therefore, in addition to mechanical trapping and aspecific adsorption of different cell types driven by DNA/histone complexes, NETs may provide specific binding sites for integrin-mediated cell adhesion of neutrophils, platelets, endothelial and cancer cells thus promoting intimate interactions among these cells. PMID:28166238

  15. Evaluation of common methods for sampling invertebrate pollinator assemblages: net sampling out-perform pan traps.

    Science.gov (United States)

    Popic, Tony J; Davila, Yvonne C; Wardle, Glenda M

    2013-01-01

    Methods for sampling ecological assemblages strive to be efficient, repeatable, and representative. Unknowingly, common methods may be limited in terms of revealing species function and so of less value for comparative studies. The global decline in pollination services has stimulated surveys of flower-visiting invertebrates, using pan traps and net sampling. We explore the relative merits of these two methods in terms of species discovery, quantifying abundance, function, and composition, and responses of species to changing floral resources. Using a spatially-nested design we sampled across a 5000 km(2) area of arid grasslands, including 432 hours of net sampling and 1296 pan trap-days, between June 2010 and July 2011. Net sampling yielded 22% more species and 30% higher abundance than pan traps, and better reflected the spatio-temporal variation of floral resources. Species composition differed significantly between methods; from 436 total species, 25% were sampled by both methods, 50% only by nets, and the remaining 25% only by pans. Apart from being less comprehensive, if pan traps do not sample flower-visitors, the link to pollination is questionable. By contrast, net sampling functionally linked species to pollination through behavioural observations of flower-visitation interaction frequency. Netted specimens are also necessary for evidence of pollen transport. Benefits of net-based sampling outweighed minor differences in overall sampling effort. As pan traps and net sampling methods are not equivalent for sampling invertebrate-flower interactions, we recommend net sampling of invertebrate pollinator assemblages, especially if datasets are intended to document declines in pollination and guide measures to retain this important ecosystem service.

  16. Production of extracellular traps against Aspergillus fumigatus in vitro and in infected lung tissue is dependent on invading neutrophils and influenced by hydrophobin RodA.

    Science.gov (United States)

    Bruns, Sandra; Kniemeyer, Olaf; Hasenberg, Mike; Aimanianda, Vishukumar; Nietzsche, Sandor; Thywissen, Andreas; Jeron, Andreas; Latgé, Jean-Paul; Brakhage, Axel A; Gunzer, Matthias

    2010-04-29

    Aspergillus fumigatus is the most important airborne fungal pathogen causing life-threatening infections in immunocompromised patients. Macrophages and neutrophils are known to kill conidia, whereas hyphae are killed mainly by neutrophils. Since hyphae are too large to be engulfed, neutrophils possess an array of extracellular killing mechanisms including the formation of neutrophil extracellular traps (NETs) consisting of nuclear DNA decorated with fungicidal proteins. However, until now NET formation in response to A. fumigatus has only been demonstrated in vitro, the importance of neutrophils for their production in vivo is unclear and the molecular mechanisms of the fungus to defend against NET formation are unknown. Here, we show that human neutrophils produce NETs in vitro when encountering A. fumigatus. In time-lapse movies NET production was a highly dynamic process which, however, was only exhibited by a sub-population of cells. NETosis was maximal against hyphae, but reduced against resting and swollen conidia. In a newly developed mouse model we could then demonstrate the existence and measure the kinetics of NET formation in vivo by 2-photon microscopy of Aspergillus-infected lungs. We also observed the enormous dynamics of neutrophils within the lung and their ability to interact with and phagocytose fungal elements in situ. Furthermore, systemic neutrophil depletion in mice almost completely inhibited NET formation in lungs, thus directly linking the immigration of neutrophils with NET formation in vivo. By using fungal mutants and purified proteins we demonstrate that hydrophobin RodA, a surface protein making conidia immunologically inert, led to reduced NET formation of neutrophils encountering Aspergillus fungal elements. NET-dependent killing of Aspergillus-hyphae could be demonstrated at later time-points, but was only moderate. Thus, these data establish that NET formation occurs in vivo during host defence against A. fumigatus, but suggest

  17. Production of extracellular traps against Aspergillus fumigatus in vitro and in infected lung tissue is dependent on invading neutrophils and influenced by hydrophobin RodA.

    Directory of Open Access Journals (Sweden)

    Sandra Bruns

    2010-04-01

    Full Text Available Aspergillus fumigatus is the most important airborne fungal pathogen causing life-threatening infections in immunocompromised patients. Macrophages and neutrophils are known to kill conidia, whereas hyphae are killed mainly by neutrophils. Since hyphae are too large to be engulfed, neutrophils possess an array of extracellular killing mechanisms including the formation of neutrophil extracellular traps (NETs consisting of nuclear DNA decorated with fungicidal proteins. However, until now NET formation in response to A. fumigatus has only been demonstrated in vitro, the importance of neutrophils for their production in vivo is unclear and the molecular mechanisms of the fungus to defend against NET formation are unknown. Here, we show that human neutrophils produce NETs in vitro when encountering A. fumigatus. In time-lapse movies NET production was a highly dynamic process which, however, was only exhibited by a sub-population of cells. NETosis was maximal against hyphae, but reduced against resting and swollen conidia. In a newly developed mouse model we could then demonstrate the existence and measure the kinetics of NET formation in vivo by 2-photon microscopy of Aspergillus-infected lungs. We also observed the enormous dynamics of neutrophils within the lung and their ability to interact with and phagocytose fungal elements in situ. Furthermore, systemic neutrophil depletion in mice almost completely inhibited NET formation in lungs, thus directly linking the immigration of neutrophils with NET formation in vivo. By using fungal mutants and purified proteins we demonstrate that hydrophobin RodA, a surface protein making conidia immunologically inert, led to reduced NET formation of neutrophils encountering Aspergillus fungal elements. NET-dependent killing of Aspergillus-hyphae could be demonstrated at later time-points, but was only moderate. Thus, these data establish that NET formation occurs in vivo during host defence against A

  18. Production of Extracellular Traps against Aspergillus fumigatus In Vitro and in Infected Lung Tissue Is Dependent on Invading Neutrophils and Influenced by Hydrophobin RodA

    Science.gov (United States)

    Aimanianda, Vishukumar; Nietzsche, Sandor; Thywißen, Andreas; Jeron, Andreas; Latgé, Jean-Paul; Brakhage, Axel A.; Gunzer, Matthias

    2010-01-01

    Aspergillus fumigatus is the most important airborne fungal pathogen causing life-threatening infections in immunocompromised patients. Macrophages and neutrophils are known to kill conidia, whereas hyphae are killed mainly by neutrophils. Since hyphae are too large to be engulfed, neutrophils possess an array of extracellular killing mechanisms including the formation of neutrophil extracellular traps (NETs) consisting of nuclear DNA decorated with fungicidal proteins. However, until now NET formation in response to A. fumigatus has only been demonstrated in vitro, the importance of neutrophils for their production in vivo is unclear and the molecular mechanisms of the fungus to defend against NET formation are unknown. Here, we show that human neutrophils produce NETs in vitro when encountering A. fumigatus. In time-lapse movies NET production was a highly dynamic process which, however, was only exhibited by a sub-population of cells. NETosis was maximal against hyphae, but reduced against resting and swollen conidia. In a newly developed mouse model we could then demonstrate the existence and measure the kinetics of NET formation in vivo by 2-photon microscopy of Aspergillus-infected lungs. We also observed the enormous dynamics of neutrophils within the lung and their ability to interact with and phagocytose fungal elements in situ. Furthermore, systemic neutrophil depletion in mice almost completely inhibited NET formation in lungs, thus directly linking the immigration of neutrophils with NET formation in vivo. By using fungal mutants and purified proteins we demonstrate that hydrophobin RodA, a surface protein making conidia immunologically inert, led to reduced NET formation of neutrophils encountering Aspergillus fungal elements. NET-dependent killing of Aspergillus-hyphae could be demonstrated at later time-points, but was only moderate. Thus, these data establish that NET formation occurs in vivo during host defence against A. fumigatus, but suggest

  19. In vivo and in vitro studies on the roles of neutrophil extracellular traps during secondary pneumococcal pneumonia after primary pulmonary influenza infection

    Directory of Open Access Journals (Sweden)

    Anandi eNarayana Moorthy

    2013-03-01

    Full Text Available Seasonal influenza virus infections may lead to debilitating disease, and account for significant fatalities annually worldwide. Most of these deaths are attributed to the complications of secondary bacterial pneumonia. Evidence is accumulating to support the notion that neutrophil extracellular traps (NETs harbor several antibacterial proteins, and trap and kill bacteria. We have previously demonstrated the induction of NETs that contribute to lung tissue injury in severe influenza pneumonia. However, the role of these NETs in secondary bacterial pneumonia is unclear. In this study, we explored whether NETs induced during pulmonary influenza infection have functional significance against infections with Streptococcus pneumoniae and other bacterial and fungal species. Our findings revealed that NETs do not participate in killing of Streptococcus pneumoniae in vivo and in vitro. Dual viral and bacterial infection elevated the bacterial load compared to animals infected with bacteria alone. Concurrently, enhanced lung pathogenesis was observed in dual-infected mice compared to those challenged with influenza virus or bacteria alone. The intensified NETs in dual-infected mice often appeared as clusters that were frequently filled with partially degraded DNA, as evidenced by punctate histone protein staining. The severe pulmonary pathology and excessive NETs generation in dual infection correlated with exaggerated inflammation and damage to the alveolar-capillary barrier. NETs stimulation in vitro did not significantly alter the gene expression of several antimicrobial proteins, and these NETs did not exhibit any bactericidal activity. Fungicidal activity against Candida albicans was observed at similar levels both in presence or absence of NETs. These results substantiate that the NETs released by primary influenza infection do not protect against secondary bacterial infection, but may compromise lung function.

  20. M1 Protein Allows Group A Streptococcal Survival in Phagocyte Extracellular Traps through Cathelicidin Inhibition

    OpenAIRE

    Lauth, Xavier; von Köckritz-Blickwede, Maren; McNamara, Case W; Myskowski, Sandra; Zinkernagel, Annelies S.; Beall, Bernard; Ghosh, Partho; Richard L Gallo; Nizet, Victor

    2009-01-01

    M1 protein contributes to Group A Streptococcus (GAS) systemic virulence by interfering with phagocytosis and through proinflammatory activities when released from the cell surface. Here we identify a novel role of M1 protein in the stimulation of neutrophil and mast cell extracellular trap formation, yet also subsequent survival of the pathogen within these DNA-based innate defense structures. Targeted mutagenesis and heterologous expression studies demonstrate M1 protein promotes resistance...

  1. Differential clearance mechanisms, neutrophil extracellular trap degradation and phagocytosis, are operative in systemic lupus erythematosus patients with distinct autoantibody specificities.

    Science.gov (United States)

    Chauhan, Sudhir Kumar; Rai, Richa; Singh, Vikas Vikram; Rai, Madhukar; Rai, Geeta

    2015-12-01

    Systemic lupus erythematosus (SLE) patients are generally presented with autoantibodies against either dsDNA or RNA-associated antigens (also known as extractable nuclear antigens, ENA) or both. However, the mechanisms and processes that lead to this distinctive autoantibody profile are not well understood. Defects in clearance mechanism i.e. phagocytosis may lead to enhanced microbial and cellular debris of immunogenic potential. In addition to defective phagocytosis, impaired neutrophil extracellular trap (NET) degradation has been recently reported in SLE patients. However, the extent to which both these clearance processes (NET-degradation and phagocytosis) are operative in serologically distinguished subsets of SLE patients is not established. Therefore, in this report, we evaluated NET-degradation and phagocytosis efficiency among SLE patients with different autoantibody specificities. SLE patients were classified into three subsets based on their autoantibody profile (anti-dsDNA, anti-ENA or both) as determined by ELISA. NET-degradation by SLE and control sera was assessed by sytox orange-based fluorescence assay. Neutrophil-mediated phagocytosis in the presence of SLE and control sera was determined by flowcytometry. The segregation of SLE patients revealed significant differences in NET-degradation and phagocytosis in SLE patients with autoantibodies against dsDNA and ENA. We report that NET-degradation efficiency was significantly impaired in SLE patients with anti-dsDNA autoantibodies and not in those with anti-ENA autoantibodies. In contrast to NET-degradation, neutrophil-mediated phagocytosis was impaired in all three subsets independent of autoantibody specificity. These observations suggest that varying clearance mechanisms are operative in SLE subsets with anti-dsDNA or anti-ENA autoantibodies. The results outlined in this manuscript also suggest that sub-grouping of SLE patients could be useful in delineating the molecular and pathological

  2. Anti-Citrullinated Protein Antibodies Are Associated With Neutrophil Extracellular Traps in the Sputum in Relatives of Rheumatoid Arthritis Patients.

    Science.gov (United States)

    Demoruelle, M Kristen; Harrall, Kylie K; Ho, Linh; Purmalek, Monica M; Seto, Nickie L; Rothfuss, Heather M; Weisman, Michael H; Solomon, Joshua J; Fischer, Aryeh; Okamoto, Yuko; Kelmenson, Lindsay B; Parish, Mark C; Feser, Marie; Fleischer, Chelsie; Anderson, Courtney; Mahler, Michael; Norris, Jill M; Kaplan, Mariana J; Cherrington, Brian D; Holers, V Michael; Deane, Kevin D

    2017-06-01

    Studies suggest that rheumatoid arthritis (RA)-related autoimmunity is initiated at a mucosal site. However, the factors associated with the mucosal generation of this autoimmunity are unknown, especially in individuals who are at risk of future RA. Therefore, we tested anti-cyclic citrullinated peptide (anti-CCP) antibodies in the sputum of RA-free first-degree relatives (FDRs) of RA patients and patients with classifiable RA. We evaluated induced sputum and serum samples from 67 FDRs and 20 RA patients for IgA anti-CCP and IgG anti-CCP, with cutoff levels for positivity determined in a control population. Sputum was also evaluated for cell counts, neutrophil extracellular traps (NETs) using sandwich enzyme-linked immunosorbent assays for protein/nucleic acid complexes, and total citrulline. Sputum was positive for IgA and/or IgG anti-CCP in 14 of 20 RA patients (70%) and 17 of 67 FDRs (25%), including a portion of FDRs who were serum anti-CCP negative. In the FDRs, elevations of sputum IgA and IgG anti-CCP were associated with elevated sputum cell counts and NET levels. IgA anti-CCP was associated with ever smoking and with elevated sputum citrulline levels. Anti-CCP is elevated in the sputum of FDRs, including seronegative FDRs, suggesting that the lung may be a site of anti-CCP generation in this population. The association of anti-CCP with elevated cell counts and NET levels in FDRs supports a hypothesis that local airway inflammation and NET formation may drive anti-CCP production in the lung and may promote the early stages of RA development. Longitudinal studies are needed to follow the evolution of these processes relative to the development of systemic autoimmunity and articular RA. © 2017, American College of Rheumatology.

  3. Antimicrobial activity of mast cells: Role and relevance of extracellular DNA traps

    Directory of Open Access Journals (Sweden)

    Helene Möllerherm

    2016-07-01

    Full Text Available Mast cells (MCs have been shown to release their nuclear DNA and subsequently form mast cell extracellular traps (MCETs comparable to neutrophil extracellular traps, which are able to entrap and kill various microbes. The formation of extracellular traps is associated with the disruption of the nuclear membrane, leads to mixing of nuclear compounds with granule components and causes the death of the cell, a process called ETosis. The question arises why do MCs release MCETs although they are very well known as multifunctional long-living sentinel cells? In the first place, MCs are known to play a role during allergic reactions and certain parasitic infections. Nonetheless, they are also critical components of the early host innate immune response to bacterial and fungal pathogens: MCs contribute to the initiation of the early immune response by recruiting effector cells including neutrophils and macrophages by locally releasing inflammatory mediators such as TNF-α. Moreover, various studies exhibit that MCs are able to eliminate microbes through intracellular as well as extracellular antimicrobial mechanisms including MCET-formation similar to that of professional phagocytes. The actual literature leads to the suggestion that MCET-formation is not the result of passive release of DNA and granule proteins during cellular disintegration, but rather an active and controlled process in response to specific stimulation which contributes to the innate host defense. This review will discuss the different known aspects of the antimicrobial activities of MCs with special focus on MCETs and their role and relevance during infection and inflammation.

  4. Antibodies mediate formation of neutrophil extracellular traps in the middle ear and facilitate secondary pneumococcal otitis media.

    Science.gov (United States)

    Short, Kirsty R; von Köckritz-Blickwede, Maren; Langereis, Jeroen D; Chew, Keng Yih; Job, Emma R; Armitage, Charles W; Hatcher, Brandon; Fujihashi, Kohtaro; Reading, Patrick C; Hermans, Peter W; Wijburg, Odilia L; Diavatopoulos, Dimitri A

    2014-01-01

    Otitis media (OM) (a middle ear infection) is a common childhood illness that can leave some children with permanent hearing loss. OM can arise following infection with a variety of different pathogens, including a coinfection with influenza A virus (IAV) and Streptococcus pneumoniae (the pneumococcus). We and others have demonstrated that coinfection with IAV facilitates the replication of pneumococci in the middle ear. Specifically, we used a mouse model of OM to show that IAV facilitates the outgrowth of S. pneumoniae in the middle ear by inducing middle ear inflammation. Here, we seek to understand how the host inflammatory response facilitates bacterial outgrowth in the middle ear. Using B cell-deficient infant mice, we show that antibodies play a crucial role in facilitating pneumococcal replication. We subsequently show that this is due to antibody-dependent neutrophil extracellular trap (NET) formation in the middle ear, which, instead of clearing the infection, allows the bacteria to replicate. We further demonstrate the importance of these NETs as a potential therapeutic target through the transtympanic administration of a DNase, which effectively reduces the bacterial load in the middle ear. Taken together, these data provide novel insight into how pneumococci are able to replicate in the middle ear cavity and induce disease.

  5. Extracellular Trapping of Soil Contaminants by Root Border Cells: New Insights into Plant Defense

    Directory of Open Access Journals (Sweden)

    Martha C. Hawes

    2016-01-01

    Full Text Available Soil and water pollution by metals and other toxic chemicals is difficult to measure and control, and, as such, presents an ongoing global threat to sustainable agriculture and human health. Efforts to remove contaminants by plant-mediated pathways, or “phytoremediation”, though widely studied, have failed to yield consistent, predictable removal of biological and chemical contaminants. Emerging research has revealed that one major limitation to using plants to clean up the environment is that plants are programmed to protect themselves: Like white blood cells in animals, border cells released from plant root tips carry out an extracellular trapping process to neutralize threats and prevent injury to the host. Variability in border cell trapping has been found to be correlated with variation in sensitivity of roots to aluminum, and removal of border cell results in increased Al uptake into the root tip. Studies now have implicated border cells in responses of diverse plant roots to a range of heavy metals, including arsenic, copper, cadmium, lead, mercury, iron, and zinc. A better understanding of border cell extracellular traps and their role in preventing toxin uptake may facilitate efforts to use plants as a nondestructive approach to neutralize environmental threats.

  6. Are sweep net sampling and pitfall trapping compatible with molecular analysis of predation?

    Science.gov (United States)

    Harwood, James D

    2008-08-01

    Molecular analysis of predation enables accurate and reliable elucidation of trophic linkages in complex food webs, but identifying the strength of such interactions can be subject to error. Currently two techniques dominate: monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR). Although the optimization and characterization of these systems ensures their sensitivity and specificity, predator collection protocols such as sweep-netting and vacuum sampling could overestimate feeding rates because of surface-level contamination, yielding positive reactivity or predation within the sampling device. Therefore, two sampling techniques (sweep-net sampling and hand collection) were compared within an alfalfa agroecosystem using a monoclonal antibody-based ELISA to test the hypothesis that cross-contamination is a source of error, i.e., significantly more predators (linyphiid spiders) would test positive for prey (Diptera) proteins. A concurrent study examining the viability of trapping predators into saline solution was also undertaken. No significant differences were found between the proportions of spiders screening positive for Diptera when collected by sweep-net versus hand collection, rejecting the hypothesis that sweep-netting predators for subsequent molecular gut content analysis overestimates predation frequency. ELISA was also capable of detecting prey proteins in predator guts from pitfall traps containing phosphate-buffered saline, indicating the suitability of this approach for the collection and analysis of epigeal predators. Although these results indicate that sweep netting and pitfall trapping into solution is appropriate in this predator-prey and ELISA analysis system, caution should be exercised with other interactions and PCR-based analysis. The likelihood for false-positive reactivity should therefore be considered on a case-by-case basis.

  7. P2Y6 Receptor Antagonist MRS2578 Inhibits Neutrophil Activation and Aggregated Neutrophil Extracellular Trap Formation Induced by Gout-Associated Monosodium Urate Crystals.

    Science.gov (United States)

    Sil, Payel; Hayes, Craig P; Reaves, Barbara J; Breen, Patrick; Quinn, Shannon; Sokolove, Jeremy; Rada, Balázs

    2017-01-01

    Human neutrophils (polymorphonuclear leukocytes [PMNs]) generate inflammatory responses within the joints of gout patients upon encountering monosodium urate (MSU) crystals. Neutrophil extracellular traps (NETs) are found abundantly in the synovial fluid of gout patients. The detailed mechanism of MSU crystal-induced NET formation remains unknown. Our goal was to shed light on possible roles of purinergic signaling and neutrophil migration in mediating NET formation induced by MSU crystals. Interaction of human neutrophils with MSU crystals was evaluated by high-throughput live imaging using confocal microscopy. We quantitated NET levels in gout synovial fluid supernatants and detected enzymatically active neutrophil primary granule enzymes, myeloperoxidase, and human neutrophil elastase. Suramin and PPADS, general P2Y receptor blockers, and MRS2578, an inhibitor of the purinergic P2Y6 receptor, blocked NET formation triggered by MSU crystals. AR-C25118925XX (P2Y2 antagonist) did not inhibit MSU crystal-stimulated NET release. Live imaging of PMNs showed that MRS2578 represses neutrophil migration and blocked characteristic formation of MSU crystal-NET aggregates called aggregated NETs. Interestingly, the store-operated calcium entry channel inhibitor (SK&F96365) also reduced MSU crystal-induced NET release. Our results indicate that the P2Y6/store-operated calcium entry/IL-8 axis is involved in MSU crystal-induced aggregated NET formation, but MRS2578 could have additional effects affecting PMN migration. The work presented in the present study could lead to a better understanding of gouty joint inflammation and help improve the treatment and care of gout patients.

  8. 中性粒细胞胞外诱捕网的研究进展%Current research development of neutrophil extracellular traps

    Institute of Scientific and Technical Information of China (English)

    张晓兰; 宋英莉; 朱辉

    2012-01-01

    中性粒细胞是体内固有免疫系统的主要成员之一,通过杀伤外来病原体来保护宿主.除了能够吞噬消灭病原体以外,中性粒细胞也能通过一种由颗粒蛋白和DNA构成的称作中性粒细胞胞外诱捕网(NETs)的结构来捕获病原体.总结近年来关于NETs的结构、产生及其生物学特性的研究进展,并设想NETs可能成为炎症和相关疾病治疗中新的研究靶点.%Neutrophils are one of the main players in the innate immune system and actively contribute to host defense by killing pathogens.Added to their ability to eliminate microorganisms by phagocytosis,neutrophils can also kill microbes by capturing them in extracellular structures consisting of granule proteins and DNA called neutrophil extracellular traps (NETs).This review summarizes the recent advancements regarding the structure,production and biological relevance of NETs.Moreover,NETs might represent a therapeutic target in inflammation and host disorders.

  9. Afa/Dr diffusely adhering Escherichia coli strain C1845 induces neutrophil extracellular traps that kill bacteria and damage human enterocyte-like cells.

    Science.gov (United States)

    Marin-Esteban, Viviana; Turbica, Isabelle; Dufour, Guillaume; Semiramoth, Nicolas; Gleizes, Aude; Gorges, Roseline; Beau, Isabelle; Servin, Alain L; Lievin-Le Moal, Vanessa; Sandré, Catherine; Chollet-Martin, Sylvie

    2012-05-01

    We recently documented the neutrophil response to enterovirulent diffusely adherent Escherichia coli expressing Afa/Dr fimbriae (Afa/Dr DAEC), using the human myeloid cell line PLB-985 differentiated into fully mature neutrophils. Upon activation, particularly during infections, neutrophils release neutrophil extracellular traps (NETs), composed of a nuclear DNA backbone associated with antimicrobial peptides, histones, and proteases, which entrap and kill pathogens. Here, using fluorescence microscopy and field emission scanning electron microscopy, we observed NET production by PLB-985 cells infected with the Afa/Dr wild-type (WT) E. coli strain C1845. We found that these NETs were able to capture, immobilize, and kill WT C1845 bacteria. We also developed a coculture model of human enterocyte-like Caco-2/TC7 cells and PLB-985 cells previously treated with WT C1845 and found, for the first time, that the F-actin cytoskeleton of enterocyte-like cells is damaged in the presence of bacterium-induced NETs and that this deleterious effect is prevented by inhibition of protease release. These findings provide new insights into the neutrophil response to bacterial infection via the production of bactericidal NETs and suggest that NETs may damage the intestinal epithelium, particularly in situations such as inflammatory bowel diseases.

  10. Factor H Binds to Extracellular DNA Traps Released from Human Blood Monocytes in Response to Candida albicans

    Science.gov (United States)

    Halder, Luke D.; Abdelfatah, Mahmoud A.; Jo, Emeraldo A. H.; Jacobsen, Ilse D.; Westermann, Martin; Beyersdorf, Niklas; Lorkowski, Stefan; Zipfel, Peter F.; Skerka, Christine

    2017-01-01

    Upon systemic infection with human pathogenic yeast Candida albicans (C. albicans), human monocytes and polymorph nuclear neutrophilic granulocytes are the first immune cells to respond and come into contact with C. albicans. Monocytes exert immediate candidacidal activity and inhibit germination, mediate phagocytosis, and kill fungal cells. Here, we show that human monocytes spontaneously respond to C. albicans cells via phagocytosis, decondensation of nuclear DNA, and release of this decondensed DNA in the form of extracellular traps (called monocytic extracellular traps: MoETs). Both subtypes of monocytes (CD14++CD16−/CD14+CD16+) formed MoETs within the first hours upon contact with C. albicans. MoETs were characterized by the presence of citrullinated histone, myeloperoxidase, lactoferrin, and elastase. MoETs were also formed in response to Staphylococcus aureus and Escherichia coli, indicating a general reaction of monocytes to infectious microbes. MoET induction differs from extracellular trap formation in macrophages as MoETs are not triggered by simvastatin, an inhibitor of cholesterol synthesis and inducer of extracellular traps in macrophages. Extracellular traps from both monocytes and neutrophils activate complement and C3b is deposited. However, factor H (FH) binds via C3b to the extracellular DNA, mediates cofactor activity, and inhibits the induction of the inflammatory cytokine interleukin-1 beta in monocytes. Altogether, the results show that human monocytes release extracellular DNA traps in response to C. albicans and that these traps finally bind FH via C3b to presumably support clearance without further inflammation. PMID:28133459

  11. Probing Intracellular Element Concentration Changes during Neutrophil Extracellular Trap Formation Using Synchrotron Radiation Based X-Ray Fluorescence

    Science.gov (United States)

    Niemiec, Maria J.; Laforce, Brecht; Garrevoet, Jan; Vergucht, Eva; De Rycke, Riet; Cloetens, Peter; Urban, Constantin F.; Vincze, Laszlo

    2016-01-01

    High pressure frozen (HPF), cryo-substituted microtome sections of 2 μm thickness containing human neutrophils (white blood cells) were analyzed using synchrotron radiation based X-ray fluorescence (SR nano-XRF) at a spatial resolution of 50 nm. Besides neutrophils from a control culture, we also analyzed neutrophils stimulated for 1–2 h with phorbol myristate acetate (PMA), a substance inducing the formation of so-called Neutrophil Extracellular Traps (or NETs), a defense system again pathogens possibly involving proteins with metal chelating properties. In order to gain insight in metal transport during this process, precise local evaluation of elemental content was performed reaching limits of detection (LODs) of 1 ppb. Mean weight fractions within entire neutrophils, their nuclei and cytoplasms were determined for the three main elements P, S and Cl, but also for the 12 following trace elements: K, Ca, Mn, Fe, Co, Ni, Cu, Zn, Se, Br, Sr and Pb. Statistical analysis, including linear regression provided objective analysis and a measure for concentration changes. The nearly linear Ca and Cl concentration changes in neutrophils could be explained by already known phenomena such as the induction of Ca channels and the uptake of Cl under activation of NET forming neutrophils. Linear concentration changes were also found for P, S, K, Mn, Fe, Co and Se. The observed linear concentration increase for Mn could be related to scavenging of this metal from the pathogen by means of the neutrophil protein calprotectin, whereas the concentration increase of Se may be related to its antioxidant function protecting neutrophils from the reactive oxygen species they produce against pathogens. We emphasize synchrotron radiation based nanoscopic X-ray fluorescence as an enabling analytical technique to study changing (trace) element concentrations throughout cellular processes, provided accurate sample preparation and data-analysis. PMID:27812122

  12. The expanding world of extracellular traps: not only neutrophils but much more

    Directory of Open Access Journals (Sweden)

    Oliver eGoldmann

    2013-01-01

    Full Text Available The release of extracellular traps (ETs is a recently described mechanism of innate immune response to infection. Although ETs have been intensely investigated in the context of neutrophil antimicrobial effector mechanisms, these structures can also be released by other immune cells such as mast cells, eosinophils and macrophages. The different ETs have several features in common, no matter from what type of cells they originated. Thus, all ETs comprise a DNA backbone with embedded antimicrobial peptides, proteases and histones. However, they also exhibit remarkable individual differences such as the subcellular compartments from where the DNA backbone originates (e.g. nucleus, mitochondria, the proportion of responding cells within the pool as well as the molecular mechanism underlying the ETs formation. This review summarizes the knowledge accumulated in recent years regarding the expanding and complex world of ETs and their role in the immune function with a particular focus on immune cells apart from neutrophils.

  13. Role of the activation gate in determining the extracellular potassium dependency of block of HERG by trapped drugs.

    Science.gov (United States)

    Pareja, Kristeen; Chu, Elaine; Dodyk, Katrina; Richter, Kristofer; Miller, Alan

    2013-01-01

    Drug induced long QT syndrome (diLQTS) results primarily from block of the cardiac potassium channel HERG (human-ether-a-go-go related gene). In some cases long QT syndrome can result in the lethal arrhythmia torsade de pointes, an arrhythmia characterized by a rapid heart rate and severely compromised cardiac output. Many patients requiring medication present with serum potassium abnormalities due to a variety of conditions including gastrointestinal dysfunction, renal and endocrine disorders, diuretic use, and aging. Extracellular potassium influences HERG channel inactivation and can alter block of HERG by some drugs. However, block of HERG by a number of drugs is not sensitive to extracellular potassium. In this study, we show that block of WT HERG by bepridil and terfenadine, two drugs previously shown to be trapped inside the HERG channel after the channel closes, is insensitive to extracellular potassium over the range of 0 mM to 20 mM. We also show that bepridil block of the HERG mutant D540K, a mutant channel that is unable to trap drugs, is dependent on extracellular potassium, correlates with the permeant ion, and is independent of HERG inactivation. These results suggest that the lack of extracellular potassium dependency of block of HERG by some drugs may in part be related to the ability of these drugs to be trapped inside the channel after the channel closes.

  14. Recognition of Aspergillus fumigatus hyphae by human plasmacytoid dendritic cells is mediated by dectin-2 and results in formation of extracellular traps.

    Directory of Open Access Journals (Sweden)

    Flávio V Loures

    2015-02-01

    Full Text Available Plasmacytoid dendritic cells (pDCs were initially considered as critical for innate immunity to viruses. However, our group has shown that pDCs bind to and inhibit the growth of Aspergillus fumigatus hyphae and that depletion of pDCs renders mice hypersusceptible to experimental aspergillosis. In this study, we examined pDC receptors contributing to hyphal recognition and downstream events in pDCs stimulated by A. fumigatus hyphae. Our data show that Dectin-2, but not Dectin-1, participates in A. fumigatus hyphal recognition, TNF-α and IFN-α release, and antifungal activity. Moreover, Dectin-2 acts in cooperation with the FcRγ chain to trigger signaling responses. In addition, using confocal and electron microscopy we demonstrated that the interaction between pDCs and A. fumigatus induced the formation of pDC extracellular traps (pETs containing DNA and citrullinated histone H3. These structures closely resembled those of neutrophil extracellular traps (NETs. The microarray analysis of the pDC transcriptome upon A. fumigatus infection also demonstrated up-regulated expression of genes associated with apoptosis as well as type I interferon-induced genes. Thus, human pDCs directly recognize A. fumigatus hyphae via Dectin-2; this interaction results in cytokine release and antifungal activity. Moreover, hyphal stimulation of pDCs triggers a distinct pattern of pDC gene expression and leads to pET formation.

  15. Of Amoebae and Men: Extracellular DNA Traps as an Ancient Cell-Intrinsic Defense Mechanism

    Science.gov (United States)

    Zhang, Xuezhi; Soldati, Thierry

    2016-01-01

    Since the discovery of the formation of DNA-based extracellular traps (ETs) by neutrophils as an innate immune defense mechanism (1), hundreds of articles describe the involvement of ETs in physiological and pathological human and animal conditions [reviewed in Ref. (2), and the previous Frontiers Research Topic on NETosis: http://www.frontiersin.org/books/NETosis_At_the_Intersection_of_Cell_Biology_Microbiology_and_Immunology/195]. Interestingly, a few reports reveal that ETs can be formed by immune cells of more ancient organisms, as far back as the common ancestor of vertebrates and invertebrates (3). Recently, we reported that the Sentinel cells of the multicellular slug of the social amoeba Dictyostelium discoideum also produce ETs to trap and kill slug-invading bacteria [see Box 1; and Figure 1 Ref. (4)]. This is a strong evidence that DNA-based cell-intrinsic defense mechanisms emerged much earlier than thought, about 1.3 billion years ago. Amazingly, using extrusion of DNA as a weapon to capture and kill uningestable microbes has its rationale. During the emergence of multicellularity, a primitive innate immune system developed in the form of a dedicated set of specialized phagocytic cells. This professionalization of immunity allowed the evolution of sophisticated defense mechanisms including the sacrifice of a small set of cells by a mechanism related to NETosis. This altruistic behavior likely emerged in steps, starting from the release of “dispensable” mitochondrial DNA by D. discoideum Sentinel cells. Grounded in this realization, one can anticipate that in the near future, many more examples of the invention and fine-tuning of ETs by early metazoan ancestors will be identified. Consequently, it can be expected that this more complete picture of the evolution of ETs will impact our views of the involvement and pathologies linked to ETs in human and animals. PMID:27458458

  16. Aflatoxin B1 Induces Reactive Oxygen Species-Mediated Autophagy and Extracellular Trap Formation in Macrophages

    Science.gov (United States)

    An, Yanan; Shi, Xiaochen; Tang, Xudong; Wang, Yang; Shen, Fengge; Zhang, Qiaoli; Wang, Chao; Jiang, Mingguo; Liu, Mingyuan; Yu, Lu

    2017-01-01

    Aflatoxins are a group of highly toxic mycotoxins with high carcinogenicity that are commonly found in foods. Aflatoxin B1 (AFB1) is the most toxic member of the aflatoxin family. A recent study reported that AFB1 can induce autophagy, but whether AFB1 can induce extracellular traps (ETs) and the relationships among innate immune responses, reactive oxygen species (ROS), and autophagy and the ETs induced by AFB1 remain unknown. Here, we demonstrated that AFB1 induced a complete autophagic process in macrophages (MΦ) (THP-1 cells and RAW264.7 cells). In addition, AFB1 induced the generation of MΦ ETs (METs) in a dose-dependent manner. In particular, the formation of METs significantly reduced the AFB1 content. Further analysis using specific inhibitors showed that the inhibition of either autophagy or ROS prevented MET formation caused by AFB1, indicating that autophagy and ROS were required for AFB1-induced MET formation. The inhibition of ROS prevented autophagy, indicating that ROS generation occurred upstream of AFB1-induced autophagy. Taken together, these data suggest that AFB1 induces ROS-mediated autophagy and ETs formation and an M1 phenotype in MΦ. PMID:28280716

  17. Neutrophil extracellular traps and bacterial biofilms in middle ear effusion of children with recurrent acute otitis media--a potential treatment target.

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    Ruth B Thornton

    Full Text Available BACKGROUND: Bacteria persist within biofilms on the middle ear mucosa of children with recurrent and chronic otitis media however the mechanisms by which these develop remain to be elucidated. Biopsies can be difficult to obtain from children and their small size limits analysis. METHODS: In this study we aimed to investigate biofilm presence in middle ear effusion (MEE from children with recurrent acute otitis media (rAOM and to determine if these may represent infectious reservoirs similarly to those on the mucosa. We examined this through culture, viability staining and fluorescent in situ hybridisation (FISH to determine bacterial species present. Most MEEs had live bacteria present using viability staining (32/36 and all effusions had bacteria present using the universal FISH probe (26/26. Of these, 70% contained 2 or more otopathogenic species. Extensive DNA stranding was also present. This DNA was largely host derived, representing neutrophil extracellular traps (NETs within which live bacteria in biofilm formations were present. When treated with the recombinant human deoxyribonuclease 1, Dornase alfa, these strands were observed to fragment. CONCLUSIONS: Bacterial biofilms, composed of multiple live otopathogenic species can be demonstrated in the MEEs of children with rAOM and that these contain extensive DNA stranding from NETs. The NETs contribute to the viscosity of the effusion, potentially contributing to its failure to clear as well as biofilm development. Our data indicates that Dornase alfa can fragment these strands and may play a role in future chronic OM treatment.

  18. d(− Lactic Acid-Induced Adhesion of Bovine Neutrophils onto Endothelial Cells Is Dependent on Neutrophils Extracellular Traps Formation and CD11b Expression

    Directory of Open Access Journals (Sweden)

    Pablo Alarcón

    2017-08-01

    Full Text Available Bovine ruminal acidosis is of economic importance as it contributes to reduced milk and meat production. This phenomenon is mainly attributed to an overload of highly fermentable carbohydrate, resulting in increased d(− lactic acid levels in serum and plasma. Ruminal acidosis correlates with elevated acute phase proteins in blood, along with neutrophil activation and infiltration into various tissues leading to laminitis and aseptic polysynovitis. Previous studies in bovine neutrophils indicated that d(− lactic acid decreased expression of L-selectin and increased expression of CD11b to concentrations higher than 6 mM, suggesting a potential role in neutrophil adhesion onto endothelia. The two aims of this study were to evaluate whether d(− lactic acid influenced neutrophil and endothelial adhesion and to trigger neutrophil extracellular trap (NET production (NETosis in exposed neutrophils. Exposure of bovine neutrophils to 5 mM d(− lactic acid elevated NET release compared to unstimulated neutrophil negative controls. Moreover, this NET contains CD11b and histone H4 citrullinated, the latter was dependent on PAD4 activation, a critical enzyme in DNA decondensation and NETosis. Furthermore, NET formation was dependent on d(− lactic acid plasma membrane transport through monocarboxylate transporter 1 (MCT1. d(− lactic acid enhanced neutrophil adhesion onto endothelial sheets as demonstrated by in vitro neutrophil adhesion assays under continuous physiological flow conditions, indicating that cell adhesion was a NET- and a CD11b/ICAM-1-dependent process. Finally, d(− lactic acid was demonstrated for the first time to trigger NETosis in a PAD4- and MCT1-dependent manner. Thus, d(− lactic acid-mediated neutrophil activation may contribute to neutrophil-derived pro-inflammatory processes, such as aseptic laminitis and/or polysynovitis in animals suffering acute ruminal acidosis.

  19. Evidence of eosinophil extracellular trap cell death in COPD: does it represent the trigger that switches on the disease?

    Science.gov (United States)

    Uribe Echevarría, Loli; Leimgruber, Carolina; García González, Jorge; Nevado, Alberto; Álvarez, Ruth; García, Luciana N; Quintar, Amado A; Maldonado, Cristina A

    2017-01-01

    In spite of the numerous studies on chronic obstructive pulmonary disease (COPD), the cellular and molecular basis of the disease’s development remain unclear. Neutrophils and eosinophils are known to be key players in COPD. Recently, neutrophil extracellular trap cell death (NETosis), a mechanism due to decondensation and extrusion of chromatin to form extracellular traps, has been demonstrated in COPD. However, there is limited knowledge about eosinophil extracellular trap cell death (EETosis) and its role in the pathogenesis of COPD. The aim of this study was to evaluate EETosis in stable COPD. Induced sputum obtained from healthy smokers and low exacerbation risk COPD A or B group patients or high exacerbation risk COPD C or D group patients were included. Samples were examined using electron microscopy and immunofluorescence. Healthy smokers (n=10) and COPD A (n=19) group exhibited neutrophilic or paucigranulocytic phenotypes, with NETosis being absent in these patients. In contrast, COPD B (n=29), with eosinophilic or mixed phenotypes, showed EETosis and incipient NETosis. COPD C (n=18) and COPD D groups (n=13) were differentiated from low exacerbation rate-COPD group by the abundant cellular debris, with COPD C group having an eosinophilic pattern and numerous cells undergoing EETosis. A hallmark of this group was the abundant released membranes that often appeared phagocytosed by neutrophils, which coincidentally exhibited early NETosis changes. The COPD D group included patients with a neutrophilic or mixed pattern, with abundant neutrophil extracellular trap-derived material. This study is the first to demonstrate EETosis at different stages of stable COPD. The results suggest a role for eosinophils in COPD pathophysiology, especially at the beginning and during the persistence of the disease, regardless of whether the patient quit smoking, with EETosis debris probably triggering uncontrolled NETosis. The main target of these findings should be young

  20. Peroxiredoxin-glutaredoxin and catalase promote resistance of nontypeable Haemophilus influenzae 86-028NP to oxidants and survival within neutrophil extracellular traps.

    Science.gov (United States)

    Juneau, Richard A; Pang, Bing; Armbruster, Chelsie E; Murrah, Kyle A; Perez, Antonia C; Swords, W Edward

    2015-01-01

    Nontypeable Haemophilus influenzae (NTHI) is a common commensal and opportunistic pathogen of the human airways. For example, NTHI is a leading cause of otitis media and is the most common cause of airway infections associated with chronic obstructive pulmonary disease (COPD). These infections are often chronic/recurrent in nature and involve bacterial persistence within biofilm communities that are highly resistant to host clearance. Our previous work has shown that NTHI within biofilms has increased expression of factors associated with oxidative stress responses. The goal of this study was to define the roles of catalase (encoded by hktE) and a bifunctional peroxiredoxin-glutaredoxin (encoded by pdgX) in resistance of NTHI to oxidants and persistence in vivo. Isogenic NTHI strain 86-028NP mutants lacking hktE and pdgX had increased susceptibility to peroxide. Moreover, these strains had persistence defects in the chinchilla infection model for otitis media, as well as in a murine model for COPD. Additional work showed that pdgX and hktE were important determinants of NTHI survival within neutrophil extracellular traps (NETs), which we have shown to be an integral part of NTHI biofilms in vivo. Based on these data, we conclude that catalase and peroxiredoxin-glutaredoxin are determinants of bacterial persistence during chronic/recurrent NTHI infections that promote bacterial survival within NETs.

  1. The use of an animal-baited net trap for collecting mosquitoes during western equine encephalitis investigations in Argentina.

    Science.gov (United States)

    Mitchell, C J; Darsie, R F; Monath, T P; Sabattini, M S; Daffner, J

    1985-03-01

    A large net trap was used to sample mosquito populations attracted to horses at three sites each in Santa Fe and Rio Negro Provinces, Argentina, during the austral summer of 1984. These provinces, as well as others in Argentina, were affected by a severe epizootic of western equine encephalitis (WEE) during 1982-83. Totals of 2,752 and 6,929 mosquitoes were collected in Santa Fe and Rio Negro Provinces during five and three trap nights, respectively. Culex mosquitoes of the subgenus Culex were predominant (45.8% of total) in the Santa Fe collections, although Aedes albifasciatus also was prevalent (21.7%). The latter species was predominant (95.7% of total) in the Rio Negro collections. The mosquito fauna was less complex (minimum of 6 species) in Rio Negro Province as compared to Santa Fe Province (minimum of 18 species). The advantages of the net trap indicate that this trap can become a useful tool in arbovirus ecology studies in other areas.

  2. 中性粒细胞胞外诱捕网蛋白质的免疫原性%Immunogenicity of proteins derived from neutrophil extracellular traps

    Institute of Scientific and Technical Information of China (English)

    周剑涛; 梅雨珍; 丁海峰

    2014-01-01

    Neutrophil extracellular traps (NETs) are composed of DNA and antimicrobial proteins. These components have diverse functions and subcellular distributions in live cells, which are redistributed and extruded from neutrophils by a death program termed NETosis. Both exuberant NETosis and impaired clearance of NETs have been implicated in the organ damage of autoimmune diseases, such as systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), Felty's syndrome (FS), small vessel vasculitis (SVV), and so on. NETs may represent an important source of neoantigens, where posttranslational modifications and proteolytic cleavage of proteins externalized in the NETs, or abnormal conformation of drug-induced NETs, could promote the generation of autoantibodies in patients with autoimmune diseases. In short, NETs may provide a unique, stimulatory microenvironment that can break normal immune tolerance, and thereby predispose to autoimmunity.%中性粒细胞胞外诱捕网(NETs)由DNA和抗菌蛋白质组成。在活细胞内,这些组分分布于亚细胞并发挥不同的功能。但是,在NETosis期,这些组分重新分布并从中性粒细胞内挤压出来。NETosis过分强盛和NETs清除受损,与自身免疫性疾病的器官损害有关,如系统性红斑狼疮(SLE)、类风湿性关节炎(RA)、Felty's综合征(FS)和小血管炎(SVV)等。NETs可能是体内新抗原的重要来源,蛋白质翻译后的修饰与水解,或药物诱导NETs构象异常能促进自身免疫性疾病患者产生自身抗体。总之, NETs可能提供了一个独特的具有破坏正常免疫耐受,引起自身免疫性的微环境。

  3. IgA Complexes in Plasma and Synovial Fluid of Patients with Rheumatoid Arthritis Induce Neutrophil Extracellular Traps via FcαRI.

    Science.gov (United States)

    Aleyd, Esil; Al, Marjon; Tuk, Cornelis W; van der Laken, Conny J; van Egmond, Marjolein

    2016-12-15

    Autoantibodies, including rheumatoid factor (RF), are an important characteristic of rheumatoid arthritis (RA). Interestingly, several studies reported a correlation between the presence of IgA autoantibodies and worse disease course. We demonstrated previously that triggering the IgA Fc receptor (FcαRI) on neutrophils results in neutrophil recruitment and the release of neutrophil extracellular traps (NETs). Because this can lead to tissue damage, we investigated whether IgA immune complexes in plasma and synovial fluid of RA patients activate neutrophils. RF isotypes were measured with ELISA, and immune complexes were precipitated using polyethylene glycol 6000. Isolated neutrophils were incubated with immune complexes, and activation and release of NETs were determined in the presence or absence of FcαRI-blocking Abs. Plasma and SF of RA patients contained IgM, IgG, and IgA RFs. Patient plasma IgA RF and IgM RF showed a strong correlation. No uptake of IgM and minimal endocytosis of IgG immune complexes by neutrophils was observed, in contrast to avid uptake of IgA complexes. Incubation of neutrophils with immune complexes resulted in the production of reactive oxygen species, as well as the release of NETs, lactoferrin, and chemotactic stimuli. Importantly, activation of neutrophils was reduced when FcαRI was blocked. Neutrophils were activated by IgA immune complexes, which suggests that neutrophils play a role in inducing joint damage in RA patients who have IgA autoantibody complexes, thereby increasing the severity of disease. Blocking FcαRI inhibited neutrophil activation and, as such, may represent an additional attractive novel therapeutic strategy for the treatment of RA.

  4. COF-Net on CNT-Net as a Molecularly Designed, Hierarchical Porous Chemical Trap for Polysulfides in Lithium-Sulfur Batteries.

    Science.gov (United States)

    Yoo, JongTae; Cho, Sung-Ju; Jung, Gwan Yeong; Kim, Su Hwan; Choi, Keun-Ho; Kim, Jeong-Hoon; Lee, Chang Kee; Kwak, Sang Kyu; Lee, Sang-Young

    2016-05-11

    The hierarchical porous structure has garnered considerable attention as a multiscale engineering strategy to bring unforeseen synergistic effects in a vast variety of functional materials. Here, we demonstrate a "microporous covalent organic framework (COF) net on mesoporous carbon nanotube (CNT) net" hybrid architecture as a new class of molecularly designed, hierarchical porous chemical trap for lithium polysulfides (Li2Sx) in Li-S batteries. As a proof of concept for the hybrid architecture, self-standing COF-net on CNT-net interlayers (called "NN interlayers") are fabricated through CNT-templated in situ COF synthesis and then inserted between sulfur cathodes and separators. Two COFs with different micropore sizes (COF-1 (0.7 nm) and COF-5 (2.7 nm)) are chosen as model systems. The effects of the pore size and (boron-mediated) chemical affinity of microporous COF nets on Li2Sx adsorption phenomena are theoretically investigated through density functional theory calculations. Benefiting from the chemical/structural uniqueness, the NN interlayers effectively capture Li2Sx without impairing their ion/electron conduction. Notably, the COF-1 NN interlayer, driven by the well-designed microporous structure, allows for the selective deposition/dissolution (i.e., facile solid-liquid conversion) of electrically inert Li2S. As a consequence, the COF-1 NN interlayer provides a significant improvement in the electrochemical performance of Li-S cells (capacity retention after 300 cycles (at charge/discharge rate = 2.0 C/2.0 C) = 84% versus 15% for a control cell with no interlayer) that lies far beyond those accessible with conventional Li-S technologies.

  5. Comparative proteomic analysis of extracellular vesicles isolated by acoustic trapping or differential centrifugation

    NARCIS (Netherlands)

    Rezeli, Melinda; Gidlöf, Olof; Evander, Mikael; Bryl-Górecka, Paulina; Sathanoori, Ramasri; Gilje, Patrik; Pawlowski, Krzysztof; Horvatovich, Péter; Erlinge, David; Marko-Varga, György; Laurell, Thomas

    2016-01-01

    Extracellular vesicles (ECVs), including microparticles (MPs) and exosomes, are submicron membrane vesicles released by diverse cell types upon activation or stress. Circulating ECVs are potential reservoirs of disease biomarkers, and the complexity of these vesicles is significantly lower compared

  6. 中性粒细胞杀灭病原体的新途径:胞外诱捕网%A new strategy for eliminating invading microbes by neutrophil: extracellular traps

    Institute of Scientific and Technical Information of China (English)

    吴腾飞; 陈福广; 黄清华; 陈伟; 邓旭明; 张巧灵; 刘波

    2013-01-01

    中性粒细胞是天然免疫系统中到达感染部位并清除微生物的第一道免疫防线的主要成员之一.已证实中性粒细胞存在3种杀灭微生物的途径:吞噬清除病原体、分泌抗微生物蛋白颗粒和新近发现的中性粒细胞诱捕网(neutrophil extracellular traps,NETs).NETs由解聚的染色体网状结构和镶嵌其中的抗微生物颗粒蛋白组成,绑定并杀灭多种微生物,包括细菌、真菌、寄生虫和病毒.中性粒细胞通过释放DNA网状结构,形成抗微生物感染的物理屏障和支架结构,限制病原的活动范围,增强抗微生物蛋白的协同作用,并减少对宿主的组织损伤.%Neutrphils as main members in the innate immune response are recruited to infection sites and form the first line of defense. Neutrophils have bee proved to employ three major antimicrobial strategies: engulfment and subsequent elimination of microbes, releasing antimicrobial molecules, and the recently described release of neutrophil extracellular traps (NETs). NETs are composed of decondensed chromatin and antimicrobial proteins, which bind and kill a variety of microbes including bacteria, parasites and fungi. The web-like NETS structure may contribute to microbial containment by forming a physical barrier and a scaffold, to enhance antimicrobial synergy while minimizing damage to host tissues.

  7. Novel anti-bacterial activities of β-defensin 1 in human platelets: suppression of pathogen growth and signaling of neutrophil extracellular trap formation.

    Directory of Open Access Journals (Sweden)

    Bjoern F Kraemer

    2011-11-01

    Full Text Available Human β-defensins (hBD are antimicrobial peptides that curb microbial activity. Although hBD's are primarily expressed by epithelial cells, we show that human platelets express hBD-1 that has both predicted and novel antibacterial activities. We observed that activated platelets surround Staphylococcus aureus (S. aureus, forcing the pathogens into clusters that have a reduced growth rate compared to S. aureus alone. Given the microbicidal activity of β-defensins, we determined whether hBD family members were present in platelets and found mRNA and protein for hBD-1. We also established that hBD-1 protein resided in extragranular cytoplasmic compartments of platelets. Consistent with this localization pattern, agonists that elicit granular secretion by platelets did not readily induce hBD-1 release. Nevertheless, platelets released hBD-1 when they were stimulated by α-toxin, a S. aureus product that permeabilizes target cells. Platelet-derived hBD-1 significantly impaired the growth of clinical strains of S. aureus. hBD-1 also induced robust neutrophil extracellular trap (NET formation by target polymorphonuclear leukocytes (PMNs, which is a novel antimicrobial function of β-defensins that was not previously identified. Taken together, these data demonstrate that hBD-1 is a previously-unrecognized component of platelets that displays classic antimicrobial activity and, in addition, signals PMNs to extrude DNA lattices that capture and kill bacteria.

  8. Research Progress of Neutrophil Extracellular Traps%中性粒细胞外网状陷阱的研究现状

    Institute of Scientific and Technical Information of China (English)

    赫嘉惠; 仲娇月; 李彩莹(综述); 赵德超(审校)

    2016-01-01

    Neutrophils are immune cells which are abundant in the human body playing a vital role in host immune defense,serving as the first line of the host defense against infection.The traditional cognition of neutrophils is more focused on its role in innate immunity.Research found that neutrophils are closely associ-ated with inflammatory immunity defense.And in recent years neutrophils were found to have another way to kill pathogens in addition to its traditional mechanisms-neutrophil extracellular traps(NETs) different from chemotaxis,phagocytosis and direct sterilization methods.Several studies have confirmed that NETs play a dual-directional regulation role on the body′s immune system.%中性粒细胞是人体内含量丰富的免疫细胞,在机体的免疫防御中占有举足轻重的地位,在机体感染的初期防御中扮演着第一道防线的重要角色。传统对中性粒细胞的认知,多集中于其在固有免疫中的作用。但研究发现,中性粒细胞与炎症免疫防御密切相关。并于近年发现了除其通过趋化、吞噬及直接杀菌方式来杀灭病原菌的传统机制外的另一种死亡方式———中性粒细胞外网状陷阱( NETs)。并证实NETs对机体免疫机制的影响起双向调节作用。

  9. A NET Outcome

    Directory of Open Access Journals (Sweden)

    Thea eLu

    2012-12-01

    Full Text Available Neutrophils constitute a critical part of innate immunity and are well known for their ability to phagocytose and kill invading microorganisms. The microbicidal processes employed by neutrophils are highly effective at killing most ingested bacteria and fungi. However, an alternative non-phagocytic antimicrobial mechanism of neutrophils has been proposed whereby microorganisms are eliminated by neutrophil extracellular traps (NETs. NETs are comprised of DNA, histones, and antimicrobial proteins extruded by neutrophils during NETosis, a cell death pathway reported to be distinct from apoptosis, phagocytosis-induced cell death, and necrosis. Although multiple laboratories have reported NETs using various stimuli in vitro, the molecular mechanisms involved in this process have yet to be definitively elucidated, and many questions regarding the formation and putative role or function of NETs in innate host defense remain unanswered. It is with these questions in mind that we provide some reflection and perspective on NETs and NETosis.

  10. Group B Streptococcus Induces Neutrophil Recruitment to Gestational Tissues and Elaboration of Extracellular Traps and Nutritional Immunity

    Science.gov (United States)

    Kothary, Vishesh; Doster, Ryan S.; Rogers, Lisa M.; Kirk, Leslie A.; Boyd, Kelli L.; Romano-Keeler, Joann; Haley, Kathryn P.; Manning, Shannon D.; Aronoff, David M.; Gaddy, Jennifer A.

    2017-01-01

    Streptococcus agalactiae, or Group B Streptococcus (GBS), is a gram-positive bacterial pathogen associated with infection during pregnancy and is a major cause of morbidity and mortality in neonates. Infection of the extraplacental membranes surrounding the developing fetus, a condition known as chorioamnionitis, is characterized histopathologically by profound infiltration of polymorphonuclear cells (PMNs, neutrophils) and greatly increases the risk for preterm labor, stillbirth, or neonatal GBS infection. The advent of animal models of chorioamnionitis provides a powerful tool to study host-pathogen relationships in vivo and ex vivo. The purpose of this study was to evaluate the innate immune response elicited by GBS and evaluate how antimicrobial strategies elaborated by these innate immune cells affect bacteria. Our work using a mouse model of GBS ascending vaginal infection during pregnancy reveals that clinically isolated GBS has the capacity to invade reproductive tissues and elicit host immune responses including infiltration of PMNs within the choriodecidua and placenta during infection, mirroring the human condition. Upon interacting with GBS, murine neutrophils elaborate DNA-containing extracellular traps, which immobilize GBS and are studded with antimicrobial molecules including lactoferrin. Exposure of GBS to holo- or apo-forms of lactoferrin reveals that the iron-sequestration activity of lactoferrin represses GBS growth and viability in a dose-dependent manner. Together, these data indicate that the mouse model of ascending infection is a useful tool to recapitulate human models of GBS infection during pregnancy. Furthermore, this work reveals that neutrophil extracellular traps ensnare GBS and repress bacterial growth via deposition of antimicrobial molecules, which drive nutritional immunity via metal sequestration strategies. PMID:28217556

  11. Far beyond Phagocytosis: Phagocyte-Derived Extracellular Traps Act Efficiently against Protozoan Parasites In Vitro and In Vivo

    Directory of Open Access Journals (Sweden)

    Liliana M. R. Silva

    2016-01-01

    Full Text Available Professional mononuclear phagocytes such as polymorphonuclear neutrophils (PMN, monocytes, and macrophages are considered as the first line of defence against invasive pathogens. The formation of extracellular traps (ETs by activated mononuclear phagocytes is meanwhile well accepted as an effector mechanism of the early host innate immune response acting against microbial infections. Recent investigations showed evidence that ETosis is a widely spread effector mechanism in vertebrates and invertebrates being utilized to entrap and kill bacteria, fungi, viruses, and protozoan parasites. ETs are released in response to intact protozoan parasites or to parasite-specific antigens in a controlled cell death process. Released ETs consist of nuclear DNA as backbone adorned with histones, antimicrobial peptides, and phagocyte-specific granular enzymes thereby producing a sticky extracellular matrix capable of entrapping and killing pathogens. This review summarizes recent data on protozoa-induced ETosis. Special attention will be given to molecular mechanisms of protozoa-induced ETosis and on its consequences for the parasites successful reproduction and life cycle accomplishment.

  12. Far beyond Phagocytosis: Phagocyte-Derived Extracellular Traps Act Efficiently against Protozoan Parasites In Vitro and In Vivo.

    Science.gov (United States)

    Silva, Liliana M R; Muñoz-Caro, Tamara; Burgos, Rafael A; Hidalgo, Maria A; Taubert, Anja; Hermosilla, Carlos

    2016-01-01

    Professional mononuclear phagocytes such as polymorphonuclear neutrophils (PMN), monocytes, and macrophages are considered as the first line of defence against invasive pathogens. The formation of extracellular traps (ETs) by activated mononuclear phagocytes is meanwhile well accepted as an effector mechanism of the early host innate immune response acting against microbial infections. Recent investigations showed evidence that ETosis is a widely spread effector mechanism in vertebrates and invertebrates being utilized to entrap and kill bacteria, fungi, viruses, and protozoan parasites. ETs are released in response to intact protozoan parasites or to parasite-specific antigens in a controlled cell death process. Released ETs consist of nuclear DNA as backbone adorned with histones, antimicrobial peptides, and phagocyte-specific granular enzymes thereby producing a sticky extracellular matrix capable of entrapping and killing pathogens. This review summarizes recent data on protozoa-induced ETosis. Special attention will be given to molecular mechanisms of protozoa-induced ETosis and on its consequences for the parasites successful reproduction and life cycle accomplishment.

  13. Trapped in the Carceral Net: Race, Gender, and the “War on Terror”

    Directory of Open Access Journals (Sweden)

    Yasmin Jiwani

    2011-01-01

    Full Text Available The events of September 11, 2001 unleashed a series of security measures designed ostensibly to protect Western nations from terrorist attacks. Many of these measures were aimed at keeping out potential terrorists and neutralizing potential terrorist activities. However, many were also aimed at citizens within the nation, legitimizing their exclusion and denial of rights. Drawing on the case of Omar Khadr, this paper argues that the carceral net ensnaring Omar Khadr was in operation well before 2001. However, since then, the carceral net has tightened to render Muslim bodies unworthy on the grounds of their putative criminality, and as undeserving victims, unbefitting state intervention and societal sympathy. The colour line, I argue, demarcates bodies that are considered worthy as opposed to those precarious others who can be penalized by the state and whose lives simply do not matter. Race, class and gender intersect and interlock to construct particular representations of victimhood as demonstrated by contemporary media representations of Muslim women.

  14. Eosinophil extracellular DNA traps in allergies = Trampas extracelulares de DNA de eosinófilos en alergias

    Directory of Open Access Journals (Sweden)

    Castrillón Álvarez, Marlon

    2014-04-01

    Full Text Available Allergy is a hypersensitivity reaction initiated by specific immunologic mechanisms. It can be mediated by antibodies or cells, developed in individuals previously sensitized by an allergen. In most cases, the antibody responsible for the allergic reaction is immunoglobulin E (IgE. Depending on the nature and mechanism of entry of the allergen, it will bring about the production of specific IgE affecting certain organs and tissues with specific symptoms. Eosinophil extracellular DNA traps or EETs are one of the mechanisms used by eosinophils during the response phase of allergy; they have not been well studied in terms of their induction, regulation and function. EETs have been detected in inflammatory intestinal processes, autoimmune diseases and multiple infectious diseases, but few investigations have been made about their involvement in allergic diseases. This is a review about the structure of EETs, the molecules involved in their formation, and their possible role in the pathogenesis of allergies. Furthermore, the main aspects of cellular and molecular processes involved in the immunopathogenesis of allergies, and the central aspects of the structure, composition and functioning of eosinophils are reviewed.

  15. Trials with portable screen rooms modified for use as animal-baited net traps for mosquito collection.

    Science.gov (United States)

    Wilton, D P; Darsie, R F; Story, R

    1985-06-01

    Trials in Larimer County, Colorado during July and August 1984, with recreational screen rooms modified as large animal-baited mosquito traps are described. The two units tested are free-standing, portable and require no external support. In all-night trials, 462.5 mosquitoes/trap night were captured with horse bait compared with 367/trap night with CDC light traps. In 2-hour evening comparisons, mosquitoes collected per trapping period totalled 416 for horse-bait traps, 132 for light traps, and 93 for human-bait traps. Animal-baited screen rooms offer an alternative to existing methods for mosquito surveillance.

  16. Myeloperoxidase associated with neutrophil extracellular traps is active and mediates bacterial killing in the presence of hydrogen peroxide.

    Science.gov (United States)

    Parker, Heather; Albrett, Amelia M; Kettle, Anthony J; Winterbourn, Christine C

    2012-03-01

    A variety of inflammatory stimuli induces NETs. These structures consist of a network of chromatin strands associated with predominately granule proteins, including MPO. NETs exhibit antimicrobial activity, which is proposed to augment the more-established mechanism of phagosomal killing. They may also be detrimental to the host in situations such as chronic inflammation or severe sepsis. The objective of this study was to establish whether MPO associated with NETs is active and able to kill bacteria. Neutrophils were stimulated with PMA to release NETs. Peroxidase activity measurements were performed and showed that enzymatically active MPO was released from the neutrophils, 2-4 h after stimulation, concomitant with NET formation. Approximately 30% of the total cellular MPO was released, with the majority bound to the NETs. The bound enzyme retained its activity. Staphylococcus aureus were not killed when added to preformed NETs under our assay conditions. However, addition of H(2)O(2) to the bacteria in the presence of NETs resulted in MPO-dependent killing, which was observed with NETs in situ and with NETs when they were removed from the neutrophils by limited DNase digestion. Our results show that the enzymatic activity of MPO on NETs could contribute to antimicrobial activity or tissue injury when NETs are released from neutrophils at sites of infection or inflammation.

  17. T Cell-Independent Mechanisms Associated with Neutrophil Extracellular Trap Formation and Selective Autophagy in IL-17A-Mediated Epidermal Hyperplasia.

    Science.gov (United States)

    Suzuki, Erika; Maverakis, Emanual; Sarin, Ritu; Bouchareychas, Laura; Kuchroo, Vijay K; Nestle, Frank O; Adamopoulos, Iannis E

    2016-12-01

    IL-17A has been strongly associated with epidermal hyperplasia in many cutaneous disorders. However, because IL-17A is mainly produced by αβ and γδT cells in response to IL-23, the role of T cells and IL-23 has overshadowed any IL-17A-independent actions. In this article, we report that IL-17A gene transfer induces epidermal hyperplasia in Il23r(-/-)Rag1(-/-)- and Tcrδ-deficient mice, which can be prevented by neutrophil depletion. Moreover, adoptive transfer of CD11b(+)Gr-1(hi) cells, after IL-17A gene transfer, was sufficient to phenocopy the disease. We further show that the IL-17A-induced pathology was prevented in transgenic mice with impaired neutrophil extracellular trap formation and/or neutrophils with conditional deletion of the master regulator of selective autophagy, Wdfy3. Our data demonstrate a novel T cell-independent mechanism that is associated with neutrophil extracellular trap formation and selective autophagy in IL-17A-mediated epidermal hyperplasia. Copyright © 2016 by The American Association of Immunologists, Inc.

  18. Gastropod-derived haemocyte extracellular traps entrap metastrongyloid larval stages of Angiostrongylus vasorum, Aelurostrongylus abstrusus and Troglostrongylus brevior

    DEFF Research Database (Denmark)

    Lange, Malin K.; Penagos-Tabares, Felipe; Munoz-Caro, Tamara

    2017-01-01

    -localization studies of haemocyte-derived extracellular DNA with histones and myeloperoxidase in larvae-entrapping structures confirmed classical characteristics of ETs. In vivo exposure of slugs to A. vasorum larvae resulted in InEPTs being extruded from haemocytes in the slug mucous extrapallial space emphasizing...... and Troglostrongylus brevior and investigated for gastropod-derived InEPT formation. Results Phase contrast as well as scanning electron microscopy (SEM) analyses of lungworm larvae-exposed haemocytes revealed ET-like structures to be extruded by haemocytes thereby contacting and ensnaring the parasites. Co...

  19. In vivo induction of cutaneous inflammation results in the accumulation of extracellular trap-forming neutrophils expressing RORγt and IL-17.

    Science.gov (United States)

    Keijsers, Romy R M C; Hendriks, Anke G M; van Erp, Piet E J; van Cranenbroek, Bram; van de Kerkhof, Peter C M; Koenen, Hans J P M; Joosten, Irma

    2014-05-01

    Clinical trials successfully using antibodies targeting IL-17 in psoriasis support the importance of IL-17 in the pathophysiology of this disease. However, there is a debate concerning the source and dynamics of IL-17 production in inflamed skin. Here we characterized IL-17-producing immune cells over time, using two established in vivo models of human skin inflammation that share many histological features with psoriasis, i.e., leukotriene B4 application and tape-stripping. Both treatments revealed a clear influx of neutrophils and T cells. Staining for IL-17 revealed that the majority of IL-17 was expressed by neutrophils and mast cells, in both models. Neutrophils, but not mast cells, coexpressed the IL-17-associated transcription factor RORγt and were able to form extracellular traps. While the presence of mast cells remained steady during the skin inflammatory process, the presence of neutrophils was clearly dynamic in time. Therefore, it is attractive to hypothesize that IL-17+/RORγt+ neutrophils contribute to human skin inflammation in vivo and possibly to the pathogenesis of skin diseases such as psoriasis. Surprisingly, T cells represented a minority of the IL-17-expressing cell population. These observations challenge the classical opinion that IL-17 is predominantly associated with T cells in skin inflammation.

  20. Bordetella parapertussis Circumvents Neutrophil Extracellular Bactericidal Mechanisms

    Science.gov (United States)

    Gorgojo, Juan; Scharrig, Emilia; Gómez, Ricardo M.; Harvill, Eric T.; Rodríguez, Maria Eugenia

    2017-01-01

    B. parapertussis is a whooping cough etiological agent with the ability to evade the immune response induced by pertussis vaccines. We previously demonstrated that in the absence of opsonic antibodies B. parapertussis hampers phagocytosis by neutrophils and macrophages and, when phagocytosed, blocks intracellular killing by interfering with phagolysosomal fusion. But neutrophils can kill and/or immobilize extracellular bacteria through non-phagocytic mechanisms such as degranulation and neutrophil extracellular traps (NETs). In this study we demonstrated that B. parapertussis also has the ability to circumvent these two neutrophil extracellular bactericidal activities. The lack of neutrophil degranulation was found dependent on the O antigen that targets the bacteria to cell lipid rafts, eventually avoiding the fusion of nascent phagosomes with specific and azurophilic granules. IgG opsonization overcame this inhibition of neutrophil degranulation. We further observed that B. parapertussis did not induce NETs release in resting neutrophils and inhibited NETs formation in response to phorbol myristate acetate (PMA) stimulation by a mechanism dependent on adenylate cyclase toxin (CyaA)-mediated inhibition of reactive oxygen species (ROS) generation. Thus, B. parapertussis modulates neutrophil bactericidal activity through two different mechanisms, one related to the lack of proper NETs-inducer stimuli and the other one related to an active inhibitory mechanism. Together with previous results these data suggest that B. parapertussis has the ability to subvert the main neutrophil bactericidal functions, inhibiting efficient clearance in non-immune hosts. PMID:28095485

  1. Proteases, neutrophils, and periodontitis: the NET effect.

    Science.gov (United States)

    Nauseef, William M

    2014-10-01

    Neutrophils exert potent antimicrobial activities in their role as first-line cellular defenders against infection. The synergistic and collective actions of oxidants and granule proteins, including serine proteases, support the microbial killing in phagosomes, where most neutrophil-mediated antimicrobial action occurs. In addition to phagocytosis, specific stimuli prompt neutrophils to extrude a matrix of DNA, histones, and granule proteins to produce neutrophil extracellular traps (NETs), which can trap microbes. Mice lacking the serine proteases necessary for NET production are more susceptible to infection, an observation suggesting that functional NETs are required for host protection. In this issue of the JCI, Sørensen and colleagues characterize neutrophils from a patient with Papillon-Lefèvre syndrome. The patient has an inactivating mutation in the gene encoding dipeptidyl peptidase I, resulting in neutrophils lacking elastase, a serine protease required for NET production. Despite the inability to form NETS, neutrophils from this patient killed pathogens in vitro, and the patient did not exhibit evidence of an increased propensity toward bacterial infections. Together, these results suggest that proteases in human neutrophils are dispensable for protection against bacterial infection and that the ability to generate NETs in vitro does not compromise host defense.

  2.  NET and NETosis – new phenomenon in immunology

    Directory of Open Access Journals (Sweden)

    Natalia Matoszka

    2012-06-01

    Full Text Available  Neutrophils are one of the first cells of the immune system recruited to the site of infection, representing the host’s most effective and numerous front-line defenders. Recently, a novel antimicrobial mechanism of neutrophils has been described: upon activation, they release DNA and a subset of their granule content, forming neutrophil extracellular traps (NETs. These extracellular, chromatin structures, which contain histones and neutrophil granule proteins, can trap and kill a broad spectrum of microbes, including Gram-positive and Gram-negative bacteria, fungi, protozoa and viruses. Some of the pathogens, which are trapped and exposed to high local concentrations of antimicrobial compounds, employ strategies against NET binding, including surface modification and/or degradation of NET by DNases. It has been suggested that NETs are formed during active cell death, recently named NETosis. New data indicate that this novel mechanism of cell death requires interaction between three processes – reactive oxygen species generation, histone citrullination and autophagy – and significantly differs from previously known types of cell death, including apoptosis and necrosis. Moreover, the release of nuclear chromatin was also described for other types of cells – mast cells and eosinophils. Mast cells, like neutrophils, under certain conditions release nuclear chromatin and may undergo a similar active cell death program, while eosinophils release only mitochondrial chromatin, and its release does not lead to the death of these cells.

  3. Ménage-à-trois: The ratio of bicarbonate to CO2 and the pH regulate the capacity of neutrophils to form NETs

    Directory of Open Access Journals (Sweden)

    Christian Maueröder

    2016-12-01

    Full Text Available In this study we identified and characterized the potential of a high ratio of bicarbonate to CO2 and a moderately alkaline pH to render neutrophils prone to undergo neutrophil extracellular trap (NET formation. Both experimental settings increased the rate of spontaneous NET release and potentiated the NET-inducing capacity of phorbol esters (PMA, ionomycin, monosodium urate and LPS. In contrast, an acidic environment impaired neutrophil extracellular trap formation both spontaneous and induced. Our findings indicate that intracellular alkalinization of neutrophils in response to an alkaline environment leads to an increase of intracellular calcium and neutrophil activation. We further found that the anion channel blocker DIDS strongly reduced NET formation induced by bicarbonate. This finding suggests that the effects observed are due to a molecular program that renders neutrophils susceptible to neutrophil extracellular trap formation. Inflammatory foci are characterized by an acidic environment. Our data indicates that NET formation is favored by the higher pH at the border regions of inflamed areas. Moreover our findings highlight the necessity for strict pH control during assays of neutrophil extracellular trap formation.

  4. NET balancing: A problem in inflammatory lung diseases

    Directory of Open Access Journals (Sweden)

    Olivia Z Cheng

    2013-01-01

    Full Text Available Neutrophil extracellular traps (NETs are beneficial antimicrobial defense structures that can help fight against invading pathogens in the host. However, recent studies reveal that NETs exert adverse effects in a number of diseases including many lung diseases. Most of the inflammatory lung diseases are characterized with a massive influx of neutrophils into the airways. Neutrophils contribute to the pathology of these diseases. To date, NETs have been identified in the lungs of cystic fibrosis, acute lung injury, allergic asthma and lungs infected with bacteria, virus, or fungi. These microbes and several host factors can stimulate NET formation, or NETosis. Different forms of NETosis have been identified; these NETotic pathways are dependent on the types of stimuli. All of these pathways however appear to result in the formation of NETs with DNA, modified extracellular histones, proteases and cytotoxic enzymes. Some of the NET components are immunogenic and damaging to the host tissue. Innate immune collectins such as pulmonary surfactant protein D (SP-D binds NETs, and enhances the clearance of dying cells and DNA by alveolar macrophages. In many inflammatory lung diseases, bronchoalveolar SP-D levels are altered and its deficiency results in the accumulation of DNA in the lungs. Some of the other therapeutic molecules under consideration for treating NET-related diseases include DNases, antiproteases, myeloperoxidase inhibitors, peptidylarginine deiminase-4 inhibitors and anti-histone antibodies. Too much of the good thing can be a bad thing. Maintaining the right balance of NET formation and reducing the amounts of NETs that accumulate in the tissues are essential for harnessing the power of NETs with minimal damage to the hosts.

  5. NET amyloidogenic backbone in human activated neutrophils.

    Science.gov (United States)

    Pulze, L; Bassani, B; Gini, E; D'Antona, P; Grimaldi, A; Luini, A; Marino, F; Noonan, D M; Tettamanti, G; Valvassori, R; de Eguileor, M

    2016-03-01

    Activated human neutrophils produce a fibrillar DNA network [neutrophil extracellular traps (NETs)] for entrapping and killing bacteria, fungi, protozoa and viruses. Our results suggest that the neutrophil extracellular traps show a resistant amyloidogenic backbone utilized for addressing reputed proteins and DNA against the non-self. The formation of amyloid fibrils in neutrophils is regulated by the imbalance of reactive oxygen species (ROS) in the cytoplasm. The intensity and source of the ROS signal is determinant for promoting stress-associated responses such as amyloidogenesis and closely related events: autophagy, exosome release, activation of the adrenocorticotrophin hormone/α-melanocyte-stimulating hormone (ACTH/α-MSH) loop and synthesis of specific cytokines. These interconnected responses in human activated neutrophils, that have been evaluated from a morphofunctional and quantitative viewpoint, represent primitive, but potent, innate defence mechanisms. In invertebrates, circulating phagocytic immune cells, when activated, show responses similar to those described previously for activated human neutrophils. Invertebrate cells within endoplasmic reticulum cisternae produce a fibrillar material which is then assembled into an amyloidogenic scaffold utilized to convey melanin close to the invader. These findings, in consideration to the critical role played by NET in the development of several pathologies, could explain the structural resistance of these scaffolds and could provide the basis for developing new diagnostic and therapeutic approaches in immunomediated diseases in which the innate branch of the immune system has a pivotal role.

  6. Extracellular entrapment and degradation of single-walled carbon nanotubes

    Science.gov (United States)

    Farrera, Consol; Bhattacharya, Kunal; Lazzaretto, Beatrice; Andón, Fernando T.; Hultenby, Kjell; Kotchey, Gregg P.; Star, Alexander; Fadeel, Bengt

    2014-05-01

    Neutrophils extrude neutrophil extracellular traps (NETs) consisting of a network of chromatin decorated with antimicrobial proteins to enable non-phagocytic killing of microorganisms. Here, utilizing a model of ex vivo activated human neutrophils, we present evidence of entrapment and degradation of carboxylated single-walled carbon nanotubes (SWCNTs) in NETs. The degradation of SWCNTs was catalyzed by myeloperoxidase (MPO) present in purified NETs and the reaction was facilitated by the addition of H2O2 and NaBr. These results show that SWCNTs can undergo acellular, MPO-mediated biodegradation and imply that the immune system may deploy similar strategies to rid the body of offending microorganisms and engineered nanomaterials.Neutrophils extrude neutrophil extracellular traps (NETs) consisting of a network of chromatin decorated with antimicrobial proteins to enable non-phagocytic killing of microorganisms. Here, utilizing a model of ex vivo activated human neutrophils, we present evidence of entrapment and degradation of carboxylated single-walled carbon nanotubes (SWCNTs) in NETs. The degradation of SWCNTs was catalyzed by myeloperoxidase (MPO) present in purified NETs and the reaction was facilitated by the addition of H2O2 and NaBr. These results show that SWCNTs can undergo acellular, MPO-mediated biodegradation and imply that the immune system may deploy similar strategies to rid the body of offending microorganisms and engineered nanomaterials. Electronic supplementary information (ESI) available: Suppl. Fig. 1 - length distribution of SWCNTs; suppl. Fig. 2 - characterization of pristine vs. oxidized SWCNTs; suppl. Fig. 3 - endotoxin evaluation; suppl. Fig. 4 - NET characterization; suppl. Fig. 5 - UV-Vis/NIR analysis of biodegradation of oxidized SWCNTs; suppl. Fig. 6 - cytotoxicity of partially degraded SWCNTs. See DOI: 10.1039/c3nr06047k

  7. Matters of life and death. How neutrophils die or survive along NET release and is "NETosis" = necroptosis?

    Science.gov (United States)

    Desai, Jyaysi; Mulay, Shrikant R; Nakazawa, Daigo; Anders, Hans-Joachim

    2016-06-01

    Neutrophil extracellular trap (NET) formation is a hallmark of many disorders that involve neutrophil recruitment, tissue damage, and inflammation. As NET formation is often associated with neutrophil death, the term "NETosis" has become popular. Upon discovery that neutrophils may survive NET release, apparent misnomers, such as "vital NETosis," have been proposed. Meanwhile, it has become obvious that certain stimuli can trigger neutrophil necroptosis, a process associated with NET-like chromatin release. Here, we discuss the relationship between NET release and neutrophil death in view highlighting that many assays used in the field do not properly distinguish between the two. An updated nomenclature is needed replacing the term "NETosis" to meet the growing variety of settings leading to chromatin release with and without neutrophil death. Dissecting which triggers of NET release involve which signaling pathway will help to define drugable molecular targets that inhibit NET release and/or neutrophil necrosis in specific disorders.

  8. Activation of PAD4 in NET formation

    Directory of Open Access Journals (Sweden)

    Amanda eRohrbach

    2012-11-01

    Full Text Available Peptidyl arginine deiminases, or PADs, convert arginine residues to the non-ribosomally encoded amino acid citrulline in a variety of protein substrates. PAD4 is expressed in granulocytes and is essential for the formation of neutrophil extracellular traps (NETs via PAD4-mediated histone citrullination. Citrullination of histones is thought to promote NET formation by inducing chromatin decondensation and facilitating the expulsion of chromosomal DNA that is coated with antimicrobial molecules. Numerous stimuli have been reported to lead to PAD4 activation and NET formation. However, how this signaling process proceeds and how PAD4 becomes activated in cells is largely unknown. Herein, we describe the various stimuli and signaling pathways that have been implicated in PAD4 activation and NET formation, including the role of reactive oxygen species generation. To provide a foundation for the above discussion, we first describe PAD4 structure and function, and how these studies led to the development of PAD-specific inhibitors. A comprehensive survey of the receptors and signaling pathways that regulate PAD4 activation will be important for our understanding of innate immunity, and the identification of signaling intermediates in PAD4 activation may also lead to the generation of pharmaceuticals to target NET-related pathogenesis.

  9. 胞外捕获器:固有免疫细胞第三种防御机制的研究进展%Research Progress on Extracellular Traps:the Third kinds of Defence Mechanisms of Innate Immune Cells

    Institute of Scientific and Technical Information of China (English)

    廖成水; Boireau Pascal; 刘明远; 程相朝

    2016-01-01

    胞外捕获器(extracellular traps,ETs)的释放是近来新发现的一种由DNA和颗粒蛋白组成的固有免疫防御机制.作为继吞噬作用、自噬之后的第三种杀伤机制,大量的研究已经证实ETs在体内外可捕获和杀伤绝大多数的细菌、病毒、真菌和寄生虫,作者对ETs的概念、形成的分子机制以及ETs在宿主防御中的作用进行了简要的介绍,这将对固有免疫细胞防御策略的新认识具有重要的意义.

  10. The circadian activity rhythm of mosquitoes through men-net-bait and CO2 lamp-trapping%入帐诱与CO2灯诱对蚊蚋种群昼夜活动节律的研究

    Institute of Scientific and Technical Information of China (English)

    党荣理; 董言德; 郑重; 郭晓霞; 张映梅; 张桂林; 赵彤言

    2011-01-01

    Objective To understand the circadian activity rhythm of mosquitoes in three environments of Beiwan region through men-net-bait and C02 lamp - trapping. Methods The circadian activity rhythm of mosquitoes through men-net-bait and C02 lamp-trapping were carried out in three different environment. Results The same result were obtained as for the species and population composition between men-net-bait and C02 lamp-trapping, and Si. Mculatum showed a little more harmful than mosquito in three environments. Aedes vexans was predominant species with a proportion of over 95% , both An. Maculatum and Ae. Caspius were very few. The same circadian activity rhythm was obtained for Aedes vexans between two methods, and the circadian activity rhythm for Si. Mculatum with a complicated activity curves were very different in three environments. Conclusion Both men-net-bait and C02 lamp - trapping are available for the research on the circadian activity rhythm of mosquitoes, further studies remain to be done with using C02 lamp-trapping to investigate the circadian activity rhythm and population for blackfly.%目的 研究人帐诱法和CO2灯诱法对北湾地区蚊、蚋种群昼夜活动的变化规律.方法 选取3种不同生境同时采用入帐诱法和CO2灯诱法进行昼夜数量动态的调查.结果 2种方法对于危害种类及组成调查结果一致,在3种生境中班布蚋的危害均大于蚊虫,刺扰伊蚊为当地的优势蚊虫,组成占到95%以上,里海伊蚊和米赛按蚊数量很少;对于种群昼夜活动节律,刺扰伊蚊2种方法结果一致,都具有晨峰和昏峰,昏峰明显,数量多,晨峰数量较少,班布蚋的活动曲线较为复杂,不同生境变化较大,但一般具有4个高峰,两种方法调查结果差异较大.结论 人帐诱和CO2灯诱都可作为蚊虫种群数量及活动节律的调查方法,但对于蚋种群CO2灯诱法是否能作为数量及活动节律的调查方法,还需做进一步的研究.

  11. Net Operations

    Institute of Scientific and Technical Information of China (English)

    蒋昌俊; 吴哲辉

    1992-01-01

    Two kinds of net operations.addition and Cartesian production of P/T nets,are introduced.They are defined on the set of underlying net of P/T systems.The conditions for preserving structural properties of Petri net after these operations are discussed.It is shown that the set of P/T nets forms and Abelian group for net addition operation and the inverse net of a P/T net in usual meaning of net theory is exactly the inverse of this P/T net as an element of the P/T net group;and that the set of P/T nets forms an Abelian ring for net addition and Caresian product operations.

  12. The Reusable Astronomy Portal (TRAP)

    Science.gov (United States)

    Donaldson, T.; Rogers, A.; Wallace, G.

    2012-09-01

    The Reusable Astronomy Portal (TRAP) aims to provide a common platform for rapidly deploying Astronomy Archives to the web. TRAP is currently under development for both the VAO Data Discovery Portal and the MAST Multi-Mission Portal (Figure 1). TRAP consists of 2 major software packages: the TRAP Client and the TRAP Server. The TRAP framework allows developers to deploy the Server, connect to data resources, then focus on building custom tools for the Client. TRAP is built upon proven industry technologies including the Ext/JS JavaScript Component Library, Mono.NET Web Services, and JSON message based APIs. The multi-layered architecture of TRAP decouples each layer: Client, Service and Data Access, enabling each to evolve independently over time. Although currently deployed to provide astronomy science data access, the TRAP architecture is flexible enough to thrive in any distributed data environment.

  13. Entamoeba histolytica induces human neutrophils to form NETs.

    Science.gov (United States)

    Ventura-Juarez, J; Campos-Esparza, Mr; Pacheco-Yepez, J; López-Blanco, J A; Adabache-Ortíz, A; Silva-Briano, M; Campos-Rodríguez, R

    2016-08-01

    Entamoeba histolytica invades the intestine and other organs during the pathogenesis of amoebiasis. In the early stages, the host organism responds with an inflammatory infiltrate composed mostly of neutrophils. It has been reported that these immune cells, activated by E. histolytica, exert a protective role by releasing proteolytic enzymes and generating reactive oxygen/nitrogen species (ROS/RNS) and antimicrobial peptides. It is now known that neutrophils also produce neutrophil extracellular traps (NETs), which are able to damage and kill pathogens. Studies have shown that intracellular protozoan pathogens, including Toxoplasma gondi, Plasmodium falciparum and Leishmania spp, induce neutrophils to release NETs and are damaged by them. However, the action of this mechanism has not been explored in relation to E. histolytica trophozoites. Through scanning electron, epifluorescence microscopy and viability assays, we show for first time that during in vitro interaction with E. histolytica trophozoites, human neutrophils released NETs that covered amoebas and reduced amoebic viability. These NETs presented histones, myeloperoxidase and decondensed chromatin. The results suggest that NETs participate in the elimination of the parasite.

  14. NET formation induced by Pseudomonas aeruginosa cystic fibrosis isolates measured as release of myeloperoxidase-DNA and neutrophil elastase-DNA complexes.

    Science.gov (United States)

    Yoo, Dae-goon; Floyd, Madison; Winn, Matthew; Moskowitz, Samuel M; Rada, Balázs

    2014-08-01

    Cystic fibrosis (CF) airway disease is characterized by Pseudomonas aeruginosa infection and recruitment of neutrophil granulocytes. Neutrophil granule components (myeloperoxidase (MPO), human neutrophil elastase (HNE)), extracellular DNA and P. aeruginosa can all be found in the CF respiratory tract and have all been associated with worsening CF lung function. Pseudomonas-induced formation of neutrophil extracellular traps (NETs) offers a likely mechanism for release of MPO, HNE and DNA from neutrophils. NETs are composed of a DNA backbone decorated with granule proteins like MPO and HNE. Here we sought to examine whether CF clinical isolates of Pseudomonas are capable of inducing NET release from human neutrophil granulocytes. We used two methods to quantify NETs. We modified a previously employed ELISA that detects MPO-DNA complexes and established a new HNE-DNA ELISA. We show that these methods reliably quantify MPO-DNA and HNE-DNA complexes, measures of NET formation. We have found that CF isolates of P. aeruginosa stimulate robust respiratory burst and NET release in human neutrophils. By comparing paired "early" and "late" bacterial isolates obtained from the same CF patient we have found that early isolates induced significantly more NET release than late isolates. Our data support that Pseudomonas-induced NET release represents an important mechanism for release of neutrophil-derived CF inflammatory mediators, and confirm that decreased induction of NET formation is required for long-term adaptation of P. aeruginosa to CF airways.

  15. Extracellular fibrils of pathogenic yeast Cryptococcus gattii are important for ecological niche, murine virulence and human neutrophil interactions.

    Directory of Open Access Journals (Sweden)

    Deborah J Springer

    Full Text Available Cryptococcus gattii, an emerging fungal pathogen of humans and animals, is found on a variety of trees in tropical and temperate regions. The ecological niche and virulence of this yeast remain poorly defined. We used Arabidopsis thaliana plants and plant-derived substrates to model C. gattii in its natural habitat. Yeast cells readily colonized scratch-wounded plant leaves and formed distinctive extracellular fibrils (40-100 nm diameter x500-3000 nm length. Extracellular fibrils were observed on live plants and plant-derived substrates by scanning electron microscopy (SEM and by high voltage- EM (HVEM. Only encapsulated yeast cells formed extracellular fibrils as a capsule-deficient C. gattii mutant completely lacked fibrils. Cells deficient in environmental sensing only formed disorganized extracellular fibrils as apparent from experiments with a C. gattii STE12alpha mutant. C. gattii cells with extracellular fibrils were more virulent in murine model of pulmonary and systemic cryptococcosis than cells lacking fibrils. C. gattii cells with extracellular fibrils were also significantly more resistant to killing by human polymorphonuclear neutrophils (PMN in vitro even though these PMN produced elaborate neutrophil extracellular traps (NETs. These observations suggest that extracellular fibril formation could be a structural adaptation of C. gattii for cell-to-cell, cell-to-substrate and/or cell-to- phagocyte communications. Such ecological adaptation of C. gattii could play roles in enhanced virulence in mammalian hosts at least initially via inhibition of host PMN- mediated killing.

  16. Neutrophil Extracellular Traps in Ulcerative Colitis

    DEFF Research Database (Denmark)

    Bennike, Tue Bjerg; Carlsen, Thomas Gelsing; Ellingsen, Torkell

    2015-01-01

    BACKGROUND: The etiology of the inflammatory bowel diseases, including ulcerative colitis (UC), remains incompletely explained. We hypothesized that an analysis of the UC colon proteome could reveal novel insights into the disease etiology. METHODS: Mucosal colon biopsies were taken by endoscopy...

  17. Dexamethasone Inhibits S. aureus-Induced Neutrophil Extracellular Pathogen-Killing Mechanism, Possibly through Toll-Like Receptor Regulation.

    Science.gov (United States)

    Wan, Ting; Zhao, Yingying; Fan, Fangli; Hu, Renjian; Jin, Xiuming

    2017-01-01

    Neutrophils release neutrophil extracellular traps (NETs) in a pathogen-killing process called NETosis. Excessive NETs formation, however, is implicated in disease pathogenesis. Therefore, to understand how NETosis is regulated, we examined the effect of dexamethasone (DXM), an anti-inflammatory drug, on this process and the role of toll-like receptors (TLRs). We stimulated human neutrophils with phorbol 12-myristate 13-acetate (PMA) or Staphylococcus aureus (S. aureus) and quantified NETs formation. We also examined the effect of DXM on the bactericidal effect of NETs and the role of reactive oxygen species (ROS) and nuclear factor (NF)-κB in DXM-regulated NETosis. DXM significantly inhibited S. aureus-induced NETosis and extracellular bacterial killing. ROS production and NF-κB activation were not involved in DXM-regulated NETosis. TLR2 and TLR4, but not TLR5 or TLR6, modified S. aureus-induced NETs formation. Neither DXM nor TLRs were involved in PMA-induced NETosis. Furthermore, TLR2 and TLR4 agonists rescued DXM-inhibited NETosis, and neither TLR2 nor TLR4 antagonists could further inhibit NETosis reduction induced by DXM, indicating that DXM may inhibit NETosis by regulating TLR2 and TLR4. In conclusion, the mechanisms of S. aureus- and PMA-induced NETosis are different. DXM decreases NETs formation independently of oxidant production and NF-κB phosphorylation and possibly via a TLR-dependent mechanism.

  18. Capability of Neutrophils to Form NETs Is Not Directly Influenced by a CMA-Targeting Peptide

    Science.gov (United States)

    Maueröder, Christian; Schall, Nicolas; Meyer, Frédéric; Mahajan, Aparna; Garnier, Benjamin; Hahn, Jonas; Kienhöfer, Deborah; Hoffmann, Markus H.; Muller, Sylviane

    2017-01-01

    During inflammatory reaction, neutrophils exhibit numerous cellular and immunological functions, notably the formation of neutrophil extracellular traps (NETs) and autophagy. NETs are composed of decondensed chromatin fibers coated with various antimicrobial molecules derived from neutrophil granules. NETs participate in antimicrobial defense and can also display detrimental roles and notably trigger some of the immune features of systemic lupus erythematosus (SLE) and other autoimmune diseases. Autophagy is a complex and finely regulated mechanism involved in the cell survival/death balance that may be connected to NET formation. To shed some light on the connection between autophagy and NET formation, we designed a number of experiments in human neutrophils and both in normal and lupus-prone MRL/lpr mice to determine whether the synthetic peptide P140, which is capable of selectively modulating chaperone-mediated autophagy (CMA) in lymphocytes, could alter NET formation. P140/Lupuzor™ is currently being evaluated in phase III clinical trials involving SLE patients. Overall our in vitro and in vivo studies established that P140 does not influence NET formation, cytokine/chemokine production, or CMA in neutrophils. Thus, the beneficial effect of P140/Lupuzor™ in SLE is apparently not directly related to modulation of neutrophil function. PMID:28191006

  19. Bigraphical Nets

    Directory of Open Access Journals (Sweden)

    Ian Mackie

    2013-02-01

    Full Text Available Interaction nets are a graphical model of computation, which has been used to define efficient evaluators for functional calculi, and specifically lambda calculi with patterns. However, the flat structure of interaction nets forces pattern matching and functional behaviour to be encoded at the same level, losing some potential parallelism. In this paper, we introduce bigraphical nets, or binets for short, as a generalisation of interaction nets using ideas from bigraphs and port graphs, and we present a formal notation and operational semantics for binets. We illustrate their expressive power by examples of applications.

  20. Net Gains

    Science.gov (United States)

    Fielker, David

    2008-01-01

    The Easter conference 2008 had several activities which for the author raised the same questions on cube nets in some work with eight-year-olds some time ago. In this article, the author muses on some problems from the Easter conference regarding nets of shapes. (Contains 1 note.)

  1. Trapping ions with lasers

    CERN Document Server

    Cormick, Cecilia; Morigi, Giovanna

    2010-01-01

    This work theoretically addresses the trapping an ionized atom with a single valence electron by means of lasers, analyzing qualitatively and quantitatively the consequences of the net charge of the particle. In our model, the coupling between the ion and the electromagnetic field includes the charge monopole and the internal dipole, within a multipolar expansion of the interaction Hamiltonian. Specifically, we perform a Power-Zienau-Woolley transformation, taking into account the motion of the center of mass. The net charge produces a correction in the atomic dipole which is of order $m_e/M$ with $m_e$ the electron mass and $M$ the total mass of the ion. With respect to neutral atoms, there is also an extra coupling to the laser field which can be approximated by that of the monopole located at the position of the center of mass. These additional effects, however, are shown to be very small compared to the dominant dipolar trapping term.

  2. Net Locality

    DEFF Research Database (Denmark)

    de Souza e Silva, Adriana Araujo; Gordon, Eric

    Provides an introduction to the new theory of Net Locality and the profound effect on individuals and societies when everything is located or locatable. Describes net locality as an emerging form of location awareness central to all aspects of digital media, from mobile phones, to Google Maps, to...... of emerging technologies, from GeoCities to GPS, Wi-Fi, Wiki Me, and Google Android....

  3. Net Locality

    DEFF Research Database (Denmark)

    de Souza e Silva, Adriana Araujo; Gordon, Eric

    Provides an introduction to the new theory of Net Locality and the profound effect on individuals and societies when everything is located or locatable. Describes net locality as an emerging form of location awareness central to all aspects of digital media, from mobile phones, to Google Maps, to...... of emerging technologies, from GeoCities to GPS, Wi-Fi, Wiki Me, and Google Android....

  4. NETs and CF Lung Disease: Current Status and Future Prospects

    Directory of Open Access Journals (Sweden)

    Robert D. Gray

    2015-01-01

    Full Text Available Cystic Fibrosis (CF is the most common fatal monogenic disease among Caucasians. While CF affects multiple organ systems, the principle morbidity arises from progressive destruction of lung architecture due to chronic bacterial infection and inflammation. It is characterized by an innate immune defect that results in colonization of the airways with bacteria such as Staphylococcus aureus and Pseudomonas aeruginosa from an early age. Within the airway microenvironment the innate immune cells including epithelial cells, neutrophils, and macrophages have all been implicated in the host defense defect. The neutrophil, however, is the principal effector cell facilitating bacterial killing, but also participates in lung damage. This is evidenced by a disproportionately elevated neutrophil burden in the airways and increased neutrophil products capable of tissue degradation, such as neutrophil elastase. The CF airways also contain an abundance of nuclear material that may be originating from neutrophils. Neutrophil extracellular traps (NETs are the product of a novel neutrophil death process that involves the expulsion of nuclear material embedded with histones, proteases, and antimicrobial proteins and peptides. NETs have been postulated to contribute to the bacterial killing capacity of neutrophils, however they also function as a source of proteases and other neutrophil products that may contribute to lung injury. Targeting nuclear material with inhaled DNase therapy improves lung function and reduces exacerbations in CF and some of these effects may be due to the degradation of NETs. We critically discuss the evidence for an antimicrobial function of NETs and their potential to cause lung damage and inflammation. We propose that CF animal models that recapitulate the human CF phenotype such as the CFTR−/− pig may be useful in further elucidating a role for NETs.

  5. RESTful NET

    CERN Document Server

    Flanders, Jon

    2008-01-01

    RESTful .NET is the first book that teaches Windows developers to build RESTful web services using the latest Microsoft tools. Written by Windows Communication Foundation (WFC) expert Jon Flanders, this hands-on tutorial demonstrates how you can use WCF and other components of the .NET 3.5 Framework to build, deploy and use REST-based web services in a variety of application scenarios. RESTful architecture offers a simpler approach to building web services than SOAP, SOA, and the cumbersome WS- stack. And WCF has proven to be a flexible technology for building distributed systems not necessa

  6. Evading the joint decision trap

    DEFF Research Database (Denmark)

    Stenbæk, Jørgen; Jensen, Mads Christian Dagnis

    2016-01-01

    is applied, including the concept of the joint-decision trap. The paper finds that the outcome was made possible by compensating the member states that were worst affected by the policy changes. A coalition of net contributors, centered on Germany and the United Kingdom, was influential regarding the overall...

  7. Trapped antihydrogen

    Science.gov (United States)

    Butler, E.; Andresen, G. B.; Ashkezari, M. D.; Baquero-Ruiz, M.; Bertsche, W.; Bowe, P. D.; Cesar, C. L.; Chapman, S.; Charlton, M.; Deller, A.; Eriksson, S.; Fajans, J.; Friesen, T.; Fujiwara, M. C.; Gill, D. R.; Gutierrez, A.; Hangst, J. S.; Hardy, W. N.; Hayden, M. E.; Humphries, A. J.; Hydomako, R.; Jenkins, M. J.; Jonsell, S.; Jørgensen, L. V.; Kemp, S. L.; Kurchaninov, L.; Madsen, N.; Menary, S.; Nolan, P.; Olchanski, K.; Olin, A.; Povilus, A.; Pusa, P.; Rasmussen, C. Ø.; Robicheaux, F.; Sarid, E.; Seif el Nasr, S.; Silveira, D. M.; So, C.; Storey, J. W.; Thompson, R. I.; van der Werf, D. P.; Wurtele, J. S.; Yamazaki, Y.

    Precision spectroscopic comparison of hydrogen and antihydrogen holds the promise of a sensitive test of the Charge-Parity-Time theorem and matter-antimatter equivalence. The clearest path towards realising this goal is to hold a sample of antihydrogen in an atomic trap for interrogation by electromagnetic radiation. Achieving this poses a huge experimental challenge, as state-of-the-art magnetic-minimum atom traps have well depths of only ˜1 T (˜0.5 K for ground state antihydrogen atoms). The atoms annihilate on contact with matter and must be `born' inside the magnetic trap with low kinetic energies. At the ALPHA experiment, antihydrogen atoms are produced from antiprotons and positrons stored in the form of non-neutral plasmas, where the typical electrostatic potential energy per particle is on the order of electronvolts, more than 104 times the maximum trappable kinetic energy. In November 2010, ALPHA published the observation of 38 antiproton annihilations due to antihydrogen atoms that had been trapped for at least 172 ms and then released—the first instance of a purely antimatter atomic system confined for any length of time (Andresen et al., Nature 468:673, 2010). We present a description of the main components of the ALPHA traps and detectors that were key to realising this result. We discuss how the antihydrogen atoms were identified and how they were discriminated from the background processes. Since the results published in Andresen et al. (Nature 468:673, 2010), refinements in the antihydrogen production technique have allowed many more antihydrogen atoms to be trapped, and held for much longer times. We have identified antihydrogen atoms that have been trapped for at least 1,000 s in the apparatus (Andresen et al., Nature Physics 7:558, 2011). This is more than sufficient time to interrogate the atoms spectroscopically, as well as to ensure that they have relaxed to their ground state.

  8. Bonding the foe – NETting neutrophils immobilize the pro-inflammatory monosodium urate crystals

    Directory of Open Access Journals (Sweden)

    Christine eSchorn

    2012-12-01

    Full Text Available In the presence of sodium, uric acid from purine metabolism precipitates as monosodium urate (MSU needles and forms renal calculi or causes gouty arthritis in kidneys and joints, respectively. The latter is characterized by red, hot and swollen arthritic joints.Here we report the in vitro effect of MSU crystals on blood granulocytes and analyse their contribution to granuloma formation and neutrophil extracellular traps (NETs formation (NETosis in synovial fluid of patients with gouty arthritis in vivo. We observed that MSU crystals induce NETosis in vitro in a reactive oxygen species (ROS-dependent manner. Indeed, blocking ROS (e.g. the oxidative burst by various antioxidants partially inhibited NETosis induced by MSU crystals. Analyses of synovial fluids and of tissue sections of patients suffering from gout revealed that NETs are also formed in vivo, especially during acute gouty flares and/or granuloma formation. Since prolonged exposure to NETs carries the risk for the development of chronic inflammation we also studied the opsonisation of NETs, as a prerequisite for their clearance. The established dead cells’ opsonins C3b, galectin-9 and CRP decorated the residual dead cells` corpses and opsonized these for disposal. Surprisingly, all three soluble pattern recognizing molecules spared the spread NET structures. We conclude that (I MSU crystals are strong inducers of ROS-dependent NETosis and (II that the prolonged presence of NET-pathogen or NET-crystal aggregates observed in patients with systemic autoimmunity, especially in those with low serum DNase-1 activity, cannot be compensated by CRP, complement and galectin mediated phagocytic clearance.

  9. Release of Active Peptidyl Arginine Deiminases by Neutrophils Can Explain Production of Extracellular Citrullinated Autoantigens in Rheumatoid Arthritis Synovial Fluid

    Science.gov (United States)

    Spengler, Julia; Lugonja, Božo; Jimmy Ytterberg, A.; Zubarev, Roman A.; Creese, Andrew J.; Pearson, Mark J.; Grant, Melissa M.; Milward, Michael; Lundberg, Karin; Buckley, Christopher D.; Filer, Andrew; Raza, Karim; Cooper, Paul R.; Chapple, Iain L.

    2015-01-01

    Objective In the majority of patients with rheumatoid arthritis (RA), antibodies specifically recognize citrullinated autoantigens that are generated by peptidylarginine deiminases (PADs). Neutrophils express high levels of PAD and accumulate in the synovial fluid (SF) of RA patients during disease flares. This study was undertaken to test the hypothesis that neutrophil cell death, induced by either NETosis (extrusion of genomic DNA–protein complexes known as neutrophil extracellular traps [NETs]) or necrosis, can contribute to production of autoantigens in the inflamed joint. Methods Extracellular DNA was quantified in the SF of patients with RA, patients with osteoarthritis (OA), and patients with psoriatic arthritis (PsA). Release of PAD from neutrophils was investigated by Western blotting, mass spectrometry, immunofluorescence staining, and PAD activity assays. PAD2 and PAD4 protein expression, as well as PAD enzymatic activity, were assessed in the SF of patients with RA and those with OA. Results Extracellular DNA was detected at significantly higher levels in RA SF than in OA SF (P < 0.001) or PsA SF (P < 0.05), and its expression levels correlated with neutrophil concentrations and PAD activity in RA SF. Necrotic neutrophils released less soluble extracellular DNA compared to NETotic cells in vitro (P < 0.05). Higher PAD activity was detected in RA SF than in OA SF (P < 0.05). The citrullinated proteins PAD2 and PAD4 were found attached to NETs and also freely diffused in the supernatant. PAD enzymatic activity was detected in supernatants of neutrophils undergoing either NETosis or necrosis. Conclusion Release of active PAD isoforms into the SF by neutrophil cell death is a plausible explanation for the generation of extracellular autoantigens in RA. PMID:26245941

  10. Trapped antihydrogen

    CERN Document Server

    Butler, E; Ashkezari, M D; Baquero-Ruiz, M; Bertsche, W; Bowe, P D; Cesar, C L; Chapman, S; Charlton, M; Deller, A; Eriksson, S; Fajans, J; Friesen, T; Fujiwara, M C; Gill, D R; Gutierrez, A; Hangst, J S; Hardy, W N; Hayden, M E; Humphries, A J; Hydomako, R; Jenkins, M J; Jonsell, S; Jørgensen, L V; Kemp, S L; Kurchaninov, L; Madsen, N; Menary, S; Nolan, P; Olchanski, K; Olin, A; Povilus, A; Pusa, P; Rasmussen, C Ø; Robicheaux, F; Sarid, E; Seif el Nasr, S; Silveira, D M; So, C; Storey, J W; Thompson, R I; van der Werf, D P; Wurtele, J S; Yamazaki,Y

    2012-01-01

    Precision spectroscopic comparison of hydrogen and antihydrogen holds the promise of a sensitive test of the Charge-Parity-Time theorem and matter-antimatter equivalence. The clearest path towards realising this goal is to hold a sample of antihydrogen in an atomic trap for interrogation by electromagnetic radiation. Achieving this poses a huge experimental challenge, as state-of-the-art magnetic-minimum atom traps have well depths of only ∼1 T (∼0.5 K for ground state antihydrogen atoms). The atoms annihilate on contact with matter and must be ‘born’ inside the magnetic trap with low kinetic energies. At the ALPHA experiment, antihydrogen atoms are produced from antiprotons and positrons stored in the form of non-neutral plasmas, where the typical electrostatic potential energy per particle is on the order of electronvolts, more than 104 times the maximum trappable kinetic energy. In November 2010, ALPHA published the observation of 38 antiproton annihilations due to antihydrogen atoms that had been ...

  11. Caught in the Net: Perineuronal Nets and Addiction

    Directory of Open Access Journals (Sweden)

    Megan Slaker

    2016-01-01

    Full Text Available Exposure to drugs of abuse induces plasticity in the brain and creates persistent drug-related memories. These changes in plasticity and persistent drug memories are believed to produce aberrant motivation and reinforcement contributing to addiction. Most studies have explored the effect drugs of abuse have on pre- and postsynaptic cells and astrocytes; however, more recently, attention has shifted to explore the effect these drugs have on the extracellular matrix (ECM. Within the ECM are unique structures arranged in a net-like manner, surrounding a subset of neurons called perineuronal nets (PNNs. This review focuses on drug-induced changes in PNNs, the molecules that regulate PNNs, and the expression of PNNs within brain circuitry mediating motivation, reward, and reinforcement as it pertains to addiction.

  12. VACUUM TRAP

    Science.gov (United States)

    Gordon, H.S.

    1959-09-15

    An improved adsorption vacuum trap for use in vacuum systems was designed. The distinguishing feature is the placement of a plurality of torsionally deformed metallic fins within a vacuum jacket extending from the walls to the central axis so that substantially all gas molecules pass through the jacket will impinge upon the fin surfaces. T fins are heated by direct metallic conduction, thereby ol taining a uniform temperature at the adeorbing surfaces so that essentially all of the condensible impurities from the evacuating gas are removed from the vacuum system.

  13. Trapped phonons

    CERN Document Server

    Mannarelli, Massimo

    2013-01-01

    We analyze the effect of restricted geometries on the contribution of Nambu-Goldstone bosons (phonons) to the shear viscosity, $\\eta$, of a superfluid. For illustrative purpose we examine a simplified system consisting of a circular boundary of radius $R$, confining a two-dimensional rarefied gas of phonons. Considering the Maxwell-type conditions, we show that phonons that are not in equilibrium with the boundary and that are not specularly reflected exert a shear stress on the boundary. In this case it is possible to define an effective (ballistic) shear viscosity coefficient $\\eta \\propto \\rho_{\\rm ph} \\chi R$, where $\\rho_{\\rm ph}$ is the density of phonons and $\\chi$ is a parameter which characterizes the type of scattering at the boundary. For an optically trapped superfluid our results corroborate the findings of Refs. \\cite{Mannarelli:2012su, Mannarelli:2012eg}, which imply that at very low temperature the shear viscosity correlates with the size of the optical trap and decreases with decreasing tempe...

  14. Extracellular DNA: the tip of root defenses?

    Science.gov (United States)

    Hawes, Martha C; Curlango-Rivera, Gilberto; Wen, Fushi; White, Gerard J; Vanetten, Hans D; Xiong, Zhongguo

    2011-06-01

    This review discusses how extracellular DNA (exDNA) might function in plant defense, and at what level(s) of innate immunity this process might operate. A new role for extracellular factors in mammalian defense has been described in a series of studies. These studies reveal that cells including neutrophils, eosinophils, and mast cells produce 'extracellular traps' (ETs) consisting of histone-linked exDNA. When pathogens are attracted to such ETs, they are trapped and killed. When the exDNA component of ETs is degraded, trapping is impaired and resistance against invasion is reduced. Conversely, mutation of microbial genes encoding exDNases that degrade exDNA results in loss of virulence. This discovery that exDNases are virulence factors opens new avenues for disease control. In plants, exDNA is required for defense of the root tip. Innate immunity-related proteins are among a group of >100 proteins secreted from the root cap and root border cell populations. Direct tests revealed that exDNA also is rapidly synthesized and exported from the root tip. When this exDNA is degraded by the endonuclease DNase 1, root tip resistance to fungal infection is lost; when the polymeric structure is degraded more slowly, by the exonuclease BAL31, loss of resistance to fungal infection is delayed accordingly. The results suggest that root border cells may function in a manner analogous to that which occurs in mammalian cells.

  15. Neutrophil Integrins and Matrix Ligands and NET Release

    Directory of Open Access Journals (Sweden)

    Jonathan S. Reichner

    2016-09-01

    Full Text Available Neutrophils are motile and responsive to tissue injury and infection. As neutrophils emigrate from the bloodstream and migrate towards a site of affliction, they encounter the tissue extracellular matrix (ECM and thereby engage integrins. Our laboratory studies the neutrophilic response to the fungal pathogen Candida albicans either in the filamentous state of the microbe or to the purified pathogen-associated molecular pattern, β-glucan. We have gained an appreciation for the role of integrins in regulating the neutrophil anti-Candida response and how the presence or absence of ECM can drive experimental outcome. The β2 integrin CR3 (Complement Receptor 3; αMβ2; Mac-1; CD11b/CD18 plays an important role in fungal recognition by its ability to bind β-glucan at a unique lectin-like domain. The presence of ECM differentially regulates essential neutrophil anti-fungal functions including chemotaxis, respiratory burst, homotypic aggregation, and the release of neutrophil extracellular traps (NETosis. We have shown that NET release to C. albicans hyphae or immobilized β-glucan occurs rapidly and without the requirement for respiratory burst on ECM. This is in contrast to the more frequently reported mechanisms of NETosis to other pathogens without the context of ECM which occur after a prolonged lag period and require respiratory burst. As expected for an ECM-dependent phenotype, NETosis and other neutrophil functions are dependent on specific β1 integrins. The focus of this review is the role of ECM ligation by neutrophil integrins as it pertains to host defense functions with an emphasis on lessons we have learned studying the anti-Candida response of human neutrophils.

  16. Annual Trapping Proposal 1985

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — This Annual Trapping Plan for the 1984-1985 trapping season at Clarence Cannon NWR outlines rules and regulations for the trapping of beaver and muskrat on the...

  17. NA-NET numerical analysis net

    Energy Technology Data Exchange (ETDEWEB)

    Dongarra, J. (Tennessee Univ., Knoxville, TN (United States). Dept. of Computer Science Oak Ridge National Lab., TN (United States)); Rosener, B. (Tennessee Univ., Knoxville, TN (United States). Dept. of Computer Science)

    1991-12-01

    This report describes a facility called NA-NET created to allow numerical analysts (na) an easy method of communicating with one another. The main advantage of the NA-NET is uniformity of addressing. All mail is addressed to the Internet host na-net.ornl.gov'' at Oak Ridge National Laboratory. Hence, members of the NA-NET do not need to remember complicated addresses or even where a member is currently located. As long as moving members change their e-mail address in the NA-NET everything works smoothly. The NA-NET system is currently located at Oak Ridge National Laboratory. It is running on the same machine that serves netlib. Netlib is a separate facility that distributes mathematical software via electronic mail. For more information on netlib consult, or send the one-line message send index'' to netlib{at}ornl.gov. The following report describes the current NA-NET system from both a user's perspective and from an implementation perspective. Currently, there are over 2100 members in the NA-NET. An average of 110 mail messages pass through this facility daily.

  18. NA-NET numerical analysis net

    Energy Technology Data Exchange (ETDEWEB)

    Dongarra, J. [Tennessee Univ., Knoxville, TN (United States). Dept. of Computer Science]|[Oak Ridge National Lab., TN (United States); Rosener, B. [Tennessee Univ., Knoxville, TN (United States). Dept. of Computer Science

    1991-12-01

    This report describes a facility called NA-NET created to allow numerical analysts (na) an easy method of communicating with one another. The main advantage of the NA-NET is uniformity of addressing. All mail is addressed to the Internet host ``na-net.ornl.gov`` at Oak Ridge National Laboratory. Hence, members of the NA-NET do not need to remember complicated addresses or even where a member is currently located. As long as moving members change their e-mail address in the NA-NET everything works smoothly. The NA-NET system is currently located at Oak Ridge National Laboratory. It is running on the same machine that serves netlib. Netlib is a separate facility that distributes mathematical software via electronic mail. For more information on netlib consult, or send the one-line message ``send index`` to netlib{at}ornl.gov. The following report describes the current NA-NET system from both a user`s perspective and from an implementation perspective. Currently, there are over 2100 members in the NA-NET. An average of 110 mail messages pass through this facility daily.

  19. Cold atom trap with zero residual magnetic field: the ac magneto-optical trap.

    Science.gov (United States)

    Harvey, Matthew; Murray, Andrew James

    2008-10-24

    A novel atom trap is described using alternating current to generate the magnetic B field, together with high speed polarization switching of the damping laser field. This combination produces a trap as effective as a standard magneto-optical trap (MOT), with the advantage that the average B field is zero. No net current is hence induced in surrounding conductive elements, and the B field produced by the ac MOT is found to switch off >300 times faster than a conventional MOT. New experiments can hence be performed, including charged particle probing or detection of the cold target ensemble.

  20. Net Ecosystem Carbon Flux

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Net Ecosystem Carbon Flux is defined as the year-over-year change in Total Ecosystem Carbon Stock, or the net rate of carbon exchange between an ecosystem and the...

  1. Shrew trap efficiency

    DEFF Research Database (Denmark)

    Gambalemoke, Mbalitini; Mukinzi, Itoka; Amundala, Drazo

    2008-01-01

    We investigated the efficiency of four trap types (pitfall, Sherman LFA, Victor snap and Museum Special snap traps) to capture shrews. This experiment was conducted in five inter-riverine forest blocks in the region of Kisangani. The total trapping effort was 6,300, 9,240, 5,280 and 5,460 trap-ni...

  2. Professional Enterprise NET

    CERN Document Server

    Arking, Jon

    2010-01-01

    Comprehensive coverage to help experienced .NET developers create flexible, extensible enterprise application code If you're an experienced Microsoft .NET developer, you'll find in this book a road map to the latest enterprise development methodologies. It covers the tools you will use in addition to Visual Studio, including Spring.NET and nUnit, and applies to development with ASP.NET, C#, VB, Office (VBA), and database. You will find comprehensive coverage of the tools and practices that professional .NET developers need to master in order to build enterprise more flexible, testable, and ext

  3. Annotating Coloured Petri Nets

    DEFF Research Database (Denmark)

    Lindstrøm, Bo; Wells, Lisa Marie

    2002-01-01

    Coloured Petri nets (CP-nets) can be used for several fundamentally different purposes like functional analysis, performance analysis, and visualisation. To be able to use the corresponding tool extensions and libraries it is sometimes necessary to include extra auxiliary information in the CP-ne...... a certain use of the CP-net. We define the semantics of annotations by describing a translation from a CP-net and the corresponding annotation layers to another CP-net where the annotations are an integrated part of the CP-net....... a method which makes it possible to associate auxiliary information, called annotations, with tokens without modifying the colour sets of the CP-net. Annotations are pieces of information that are not essential for determining the behaviour of the system being modelled, but are rather added to support...

  4. Evaluation of capture techniques on lesser prairie-chicken trap injury and survival

    Science.gov (United States)

    Grisham, Blake A.; Boal, Clint W.; Mitchell, Natasia R.; Gicklhorn, Trevor S.; Borsdorf, Philip K.; Haukos, David A.; Dixon, Charles

    2015-01-01

    Ethical treatment of research animals is required under the Animal Welfare Act. This includes trapping methodologies that reduce unnecessary pain and duress. Traps used in research should optimize animal welfare conditions within the context of the proposed research study. Several trapping techniques are used in the study of lesser prairie-chickens, despite lack of knowledge of trap injury caused by the various methods. From 2006 to 2012, we captured 217, 40, and 144 lesser prairie-chickens Tympanuchus pallidicinctus using walk-in funnel traps, rocket nets, and drop nets, respectively, in New Mexico and Texas, to assess the effects of capture technique on injury and survival of the species. We monitored radiotagged, injured lesser prairie-chickens 7–65 d postcapture to assess survival rates of injured individuals. Injuries occurred disproportionately among trap type, injury type, and sex. The predominant injuries were superficial cuts to the extremities of males captured in walk-in funnel traps. However, we observed no mortalities due to trapping, postcapture survival rates of injured birds did not vary across trap types, and the daily survival probability of an injured and uninjured bird was ≥99%. Frequency and intensity of injuries in walk-in funnel traps are due to the passive nature of these traps (researcher cannot select specific individuals for capture) and incidental capture of individuals not needed for research. Comparatively, rocket nets and drop nets allow observers to target birds for capture and require immediate removal of captured individuals from the trap. Based on our results, trap injuries would be reduced if researchers monitor and immediately remove birds from walk-in funnels before they injure themselves; move traps to target specific birds and reduce recaptures; limit the number of consecutive trapping days on a lek; and use proper netting techniques that incorporate quick, efficient, trained handling procedures.

  5. Phylogenic analysis of adhesion related genes Mad1 revealed a positive selection for the evolution of trapping devices of nematode-trapping fungi.

    Science.gov (United States)

    Li, Juan; Liu, Yue; Zhu, Hongyan; Zhang, Ke-Qin

    2016-03-04

    Adhesions, the major components of the extracellular fibrillar polymers which accumulate on the outer surface of adhesive traps of nematode-trapping fungi, are thought to have played important roles during the evolution of trapping devices. Phylogenetic analyses based on the genes related to adhesive materials can be of great importance for understanding the evolution of trapping devices. Recently, AoMad1, one homologous gene of the entomopathogenic fungus Metarhizium anisopliae cell wall protein MAD1, has been functionally characterized as involved in the production of adhesions in the nematode-trapping fungus Arthrobotrys oligospora. In this study, we cloned Mad1 homologous genes from nematode-trapping fungi with various trapping devices. Phylogenetic analyses suggested that species which formed nonadhesive constricting ring (CR) traps more basally placed and species with adhesive traps evolved along two lineages. Likelihood ratio tests (LRT) revealed that significant positive selective pressure likely acted on the ancestral trapping devices including both adhesive and mechanical traps, indicating that the Mad1 genes likely played important roles during the evolution of nematode-trapping fungi. Our study provides new insights into the evolution of trapping devices of nematode-trapping fungi and also contributes to understanding the importance of adhesions during the evolution of nematode-trapping fungi.

  6. Coloured Petri Nets

    DEFF Research Database (Denmark)

    Jensen, Kurt

    1991-01-01

    This paper describes how Coloured Petri Nets (CP-nets) have been developed — from being a promising theoretical model to being a full-fledged language for the design, specification, simulation, validation and implementation of large software systems (and other systems in which human beings and...... use of CP-nets — because it means that the function representation and the translations (which are a bit mathematically complex) no longer are parts of the basic definition of CP-nets. Instead they are parts of the invariant method (which anyway demands considerable mathematical skills...

  7. Programming NET Web Services

    CERN Document Server

    Ferrara, Alex

    2007-01-01

    Web services are poised to become a key technology for a wide range of Internet-enabled applications, spanning everything from straight B2B systems to mobile devices and proprietary in-house software. While there are several tools and platforms that can be used for building web services, developers are finding a powerful tool in Microsoft's .NET Framework and Visual Studio .NET. Designed from scratch to support the development of web services, the .NET Framework simplifies the process--programmers find that tasks that took an hour using the SOAP Toolkit take just minutes. Programming .NET

  8. Game Coloured Petri Nets

    DEFF Research Database (Denmark)

    Westergaard, Michael

    2006-01-01

    This paper introduces the notion of game coloured Petri nets. This allows the modeler to explicitly model what parts of the model comprise the modeled system and what parts are the environment of the modeled system. We give the formal definition of game coloured Petri nets, a means of reachability...... analysis of this net class, and an application of game coloured Petri nets to automatically generate easy-to-understand visualizations of the model by exploiting the knowledge that some parts of the model are not interesting from a visualization perspective (i.e. they are part of the environment...

  9. The Chemorepulsive Protein Semaphorin 3A and Perineuronal Net-Mediated Plasticity

    NARCIS (Netherlands)

    de Winter, F; Kwok, J C F; Fawcett, J W; Vo, T T; Carulli, D; Verhaagen, J

    2016-01-01

    During postnatal development, closure of critical periods coincides with the appearance of extracellular matrix structures, called perineuronal nets (PNN), around various neuronal populations throughout the brain. The absence or presence of PNN strongly correlates with neuronal plasticity. It is not

  10. Extracellular calcium sensing and extracellular calcium signaling

    Science.gov (United States)

    Brown, E. M.; MacLeod, R. J.; O'Malley, B. W. (Principal Investigator)

    2001-01-01

    The cloning of a G protein-coupled extracellular Ca(2+) (Ca(o)(2+))-sensing receptor (CaR) has elucidated the molecular basis for many of the previously recognized effects of Ca(o)(2+) on tissues that maintain systemic Ca(o)(2+) homeostasis, especially parathyroid chief cells and several cells in the kidney. The availability of the cloned CaR enabled the development of DNA and antibody probes for identifying the CaR's mRNA and protein, respectively, within these and other tissues. It also permitted the identification of human diseases resulting from inactivating or activating mutations of the CaR gene and the subsequent generation of mice with targeted disruption of the CaR gene. The characteristic alterations in parathyroid and renal function in these patients and in the mice with "knockout" of the CaR gene have provided valuable information on the CaR's physiological roles in these tissues participating in mineral ion homeostasis. Nevertheless, relatively little is known about how the CaR regulates other tissues involved in systemic Ca(o)(2+) homeostasis, particularly bone and intestine. Moreover, there is evidence that additional Ca(o)(2+) sensors may exist in bone cells that mediate some or even all of the known effects of Ca(o)(2+) on these cells. Even more remains to be learned about the CaR's function in the rapidly growing list of cells that express it but are uninvolved in systemic Ca(o)(2+) metabolism. Available data suggest that the receptor serves numerous roles outside of systemic mineral ion homeostasis, ranging from the regulation of hormonal secretion and the activities of various ion channels to the longer term control of gene expression, programmed cell death (apoptosis), and cellular proliferation. In some cases, the CaR on these "nonhomeostatic" cells responds to local changes in Ca(o)(2+) taking place within compartments of the extracellular fluid (ECF) that communicate with the outside environment (e.g., the gastrointestinal tract). In others

  11. Small Mammal Trapping 2003

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — Small mammal traps were placed in the Baring division and in the Edmunds division of Moosehom National Wildlife Refuge. There were a total of 98 traps set for up to...

  12. St. Croix trap study

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — The data set contains detailed information about the catch from 600 trap stations around St. Croix. Data fields include species caught, size data, trap location...

  13. Global Liquidity Trap

    OpenAIRE

    Fujiwara, Ippei; NAKAJIMA Tomoyuki; Sudo, Nao; Teranishi, Yuki

    2011-01-01

    In this paper we consider a two-country New Open Economy Macroeconomics model, and analyze the optimal monetary policy when countries cooperate in the face of a "global liquidity trap" -- i.e., a situation where the two countries are simultaneously caught in liquidity traps. The notable features of the optimal policy in the face of a global liquidity trap are history dependence and international dependence. The optimality of history dependent policy is confirmed as in local liquidity trap. A ...

  14. Ion trap simulation tools.

    Energy Technology Data Exchange (ETDEWEB)

    Hamlet, Benjamin Roger

    2009-02-01

    Ion traps present a potential architecture for future quantum computers. These computers are of interest due to their increased power over classical computers stemming from the superposition of states and the resulting capability to simultaneously perform many computations. This paper describes a software application used to prepare and visualize simulations of trapping and maneuvering ions in ion traps.

  15. Kunstige neurale net

    DEFF Research Database (Denmark)

    Hørning, Annette

    1994-01-01

    Artiklen beskæftiger sig med muligheden for at anvende kunstige neurale net i forbindelse med datamatisk procession af naturligt sprog, specielt automatisk talegenkendelse.......Artiklen beskæftiger sig med muligheden for at anvende kunstige neurale net i forbindelse med datamatisk procession af naturligt sprog, specielt automatisk talegenkendelse....

  16. SolNet

    DEFF Research Database (Denmark)

    Jordan, Ulrike; Vajen, Klaus; Bales, Chris;

    2014-01-01

    SolNet, founded in 2006, is the first coordinated International PhD education program on Solar Thermal Engineering. The SolNet network is coordinated by the Institute of Thermal Engineering at Kassel University, Germany. The network offers PhD courses on solar heating and cooling, conference...

  17. Net zero water

    CSIR Research Space (South Africa)

    Lindeque, M

    2013-01-01

    Full Text Available Is it possible to develop a building that uses a net zero amount of water? In recent years it has become evident that it is possible to have buildings that use a net zero amount of electricity. This is possible when the building is taken off...

  18. Net Zero Energy Buildings

    DEFF Research Database (Denmark)

    Marszal, Anna Joanna; Bourrelle, Julien S.; Musall, Eike

    2010-01-01

    and identify possible renewable energy supply options which may be considered in calculations. Finally, the gap between the methodology proposed by each organisation and their respective national building code is assessed; providing an overview of the possible changes building codes will need to undergo......The international cooperation project IEA SHC Task 40 / ECBCS Annex 52 “Towards Net Zero Energy Solar Buildings”, attempts to develop a common understanding and to set up the basis for an international definition framework of Net Zero Energy Buildings (Net ZEBs). The understanding of such buildings...... and how the Net ZEB status should be calculated differs in most countries. This paper presents an overview of Net ZEBs energy calculation methodologies proposed by organisations representing eight different countries: Austria, Canada, Denmark, Germany, Italy, Norway, Switzerland and the USA. The different...

  19. Superconducting microfabricated ion traps

    CERN Document Server

    Wang, Shannon X; Labaziewicz, Jaroslaw; Dauler, Eric; Berggren, Karl; Chuang, Isaac L

    2010-01-01

    We fabricate superconducting ion traps with niobium and niobium nitride and trap single 88Sr ions at cryogenic temperatures. The superconducting transition is verified and characterized by measuring the resistance and critical current using a 4-wire measurement on the trap structure, and observing change in the rf reflection. The lowest observed heating rate is 2.1(3) quanta/sec at 800 kHz at 6 K and shows no significant change across the superconducting transition, suggesting that anomalous heating is primarily caused by noise sources on the surface. This demonstration of superconducting ion traps opens up possibilities for integrating trapped ions and molecular ions with superconducting devices.

  20. Pro NET Best Practices

    CERN Document Server

    Ritchie, Stephen D

    2011-01-01

    Pro .NET Best Practices is a practical reference to the best practices that you can apply to your .NET projects today. You will learn standards, techniques, and conventions that are sharply focused, realistic and helpful for achieving results, steering clear of unproven, idealistic, and impractical recommendations. Pro .NET Best Practices covers a broad range of practices and principles that development experts agree are the right ways to develop software, which includes continuous integration, automated testing, automated deployment, and code analysis. Whether the solution is from a free and

  1. Getting to Net Zero

    Energy Technology Data Exchange (ETDEWEB)

    2016-09-01

    The technology necessary to build net zero energy buildings (NZEBs) is ready and available today, however, building to net zero energy performance levels can be challenging. Energy efficiency measures, onsite energy generation resources, load matching and grid interaction, climatic factors, and local policies vary from location to location and require unique methods of constructing NZEBs. It is recommended that Components start looking into how to construct and operate NZEBs now as there is a learning curve to net zero construction and FY 2020 is just around the corner.

  2. Net Zero Energy Buildings

    DEFF Research Database (Denmark)

    Marszal, Anna Joanna; Bourrelle, Julien S.; Musall, Eike

    2010-01-01

    The international cooperation project IEA SHC Task 40 / ECBCS Annex 52 “Towards Net Zero Energy Solar Buildings”, attempts to develop a common understanding and to set up the basis for an international definition framework of Net Zero Energy Buildings (Net ZEBs). The understanding of such buildings...... and identify possible renewable energy supply options which may be considered in calculations. Finally, the gap between the methodology proposed by each organisation and their respective national building code is assessed; providing an overview of the possible changes building codes will need to undergo...

  3. Extracellular Gd-CA

    DEFF Research Database (Denmark)

    Thomsen, Henrik S; Marckmann, Peter

    2008-01-01

    Until recently it was believed that extracellular gadolinium-based contrast agents were safe for both the kidneys and all other organs within the dose range up to 0.3 mmol/kg body weight. However, in 2006, it was demonstrated that some gadolinium-based contrast agents may trig the development of ...

  4. Expansion Nets and Expansion Processes of Elementary Net Systems

    Institute of Scientific and Technical Information of China (English)

    曹存根

    1995-01-01

    Occurrence nets are insufficient to precisely describe executions of elementary net systems with contacts.Traditionally,S-complementation is used for removal of contacts from the systems.Although the main behavior and properties of the original elementary net systems are preserved during S-complementation,their topologies may be changed greatly.This paper introduces a new kind of nets-expansion nets-for representing behavior of elementary net systems.As shown in the paper,expansion nets are natural as well as sufficient for describing the precise behavior of elementary net systems with or without contactks.

  5. PhysioNet

    Data.gov (United States)

    U.S. Department of Health & Human Services — The PhysioNet Resource is intended to stimulate current research and new investigations in the study of complex biomedical and physiologic signals. It offers free...

  6. Trap style influences wild pig behavior and trapping success

    Science.gov (United States)

    Williams, B.L.; Holtfreter, R.W.; Ditchkoff, S.S.; Grand, J.B.

    2011-01-01

    Despite the efforts of many natural resource professionals, wild pig (Sus scrofa) populations are expanding in many areas of the world. Although many creative techniques for controlling pig populations are being explored, trapping has been and still is themost commonly usedmethod of population control formany public and private land managers. We conducted an observational study to examine the efficiency of 2 frequently used trap styles: a small, portable box-style trap and a larger, semi-permanent, corral-style trap.We used game cameras to examine patterns of trap entry by wild pigs around each style of trap, and we conducted a trapping session to compare trapping success between trap styles. Adult female and juvenile wild pigs entered both styles of trap more readily than did adult males, and adult males seemed particularly averse to entering box traps. Less than 10% of adult male visits to box traps resulted in entries, easily the least percentage of any class at any style of trap. Adult females entered corral traps approximately 2.2 times more often per visit than box traps and re-entered corral traps >2 times more frequently. Juveniles entered and reentered both box and corral traps at similar rates. Overall (all-class) entry-per-visit rates at corral traps (0.71) were nearly double that of box traps (0.37). Subsequent trapping data supported these preliminary entry data; the capture rate for corral traps was >4 times that of box traps. Our data suggest that corral traps are temporally and economically superior to box traps with respect to efficiency; that is, corral traps effectively trap more pigs per trap night at a lower cost per pig than do box traps. ?? 2011 The Wildlife Society.

  7. Programming NET 35

    CERN Document Server

    Liberty, Jesse

    2009-01-01

    Bestselling author Jesse Liberty and industry expert Alex Horovitz uncover the common threads that unite the .NET 3.5 technologies, so you can benefit from the best practices and architectural patterns baked into the new Microsoft frameworks. The book offers a Grand Tour" of .NET 3.5 that describes how the principal technologies can be used together, with Ajax, to build modern n-tier and service-oriented applications. "

  8. Game Coloured Petri Nets

    DEFF Research Database (Denmark)

    Westergaard, Michael

    2006-01-01

    This paper introduces the notion of game coloured Petri nets. This allows the modeler to explicitly model what parts of the model comprise the modeled system and what parts are the environment of the modeled system. We give the formal definition of game coloured Petri nets, a means of reachabilit......, and not controllable by the system itself, or they are part of the system itself and therefore we need not worry about them)....

  9. Microfabricated ion trap array

    Science.gov (United States)

    Blain, Matthew G.; Fleming, James G.

    2006-12-26

    A microfabricated ion trap array, comprising a plurality of ion traps having an inner radius of order one micron, can be fabricated using surface micromachining techniques and materials known to the integrated circuits manufacturing and microelectromechanical systems industries. Micromachining methods enable batch fabrication, reduced manufacturing costs, dimensional and positional precision, and monolithic integration of massive arrays of ion traps with microscale ion generation and detection devices. Massive arraying enables the microscale ion traps to retain the resolution, sensitivity, and mass range advantages necessary for high chemical selectivity. The reduced electrode voltage enables integration of the microfabricated ion trap array with on-chip circuit-based rf operation and detection electronics (i.e., cell phone electronics). Therefore, the full performance advantages of the microfabricated ion trap array can be realized in truly field portable, handheld microanalysis systems.

  10. Vascular endothelial growth factor trap-eye and trap technology: Aflibercept from bench to bedside.

    Science.gov (United States)

    Al-Halafi, Ali M

    2014-09-01

    Anti-vascular endothelial growth factor (VEGF) currently used to treat eye diseases have included monoclonal antibodies, antibody fragments, and an aptamer. A different method of achieving VEGF blockade in retinal diseases includes the concept of a cytokine trap. Cytokine traps technology are being evaluated for the treatment of various diseases that are driven by excessive cytokine levels. Traps consist of two extracellular cytokine receptor domains fused together to form a human immunoglobulin G (IgG). Aflibercept/VEGF trap-eye (VTE) is a soluble fusion protein, which combines ligand-binding elements taken from the extracellular components of VEGF receptors 1 and 2 fused to the Fc portion of IgG. This protein contains all human amino acid sequences, which minimizes the potential for immunogenicity in human patients. This review presents the latest data on VTE in regard to the pharmacokinetics, dosage and safety, preclinical and clinical experiences. Method of the literature search: A systematic search of the literature was conducted on PubMed, Scopus, and Google Scholar with no limitation on language or year of publication databases. It was oriented to articles published for VTE in preclinical and clinical studies and was focused on the pharmacokinetics, dosage and safety of VTE.

  11. Neutral atom traps.

    Energy Technology Data Exchange (ETDEWEB)

    Pack, Michael Vern

    2008-12-01

    This report describes progress in designing a neutral atom trap capable of trapping sub millikelvin atom in a magnetic trap and shuttling the atoms across the atom chip from a collection area to an optical cavity. The numerical simulation and atom chip design are discussed. Also, discussed are preliminary calculations of quantum noise sources in Kerr nonlinear optics measurements based on electromagnetically induced transparency. These types of measurements may be important for quantum nondemolition measurements at the few photon limit.

  12. Towards trapped antihydrogen

    Science.gov (United States)

    Jørgensen, L. V.; Andresen, G.; Bertsche, W.; Boston, A.; Bowe, P. D.; Cesar, C. L.; Chapman, S.; Charlton, M.; Fajans, J.; Fujiwara, M. C.; Funakoshi, R.; Gill, D. R.; Hangst, J. S.; Hayano, R. S.; Hydomako, R.; Jenkins, M. J.; Kurchaninov, L.; Madsen, N.; Nolan, P.; Olchanski, K.; Olin, A.; Page, R. D.; Povilus, A.; Robicheaux, F.; Sarid, E.; Silveira, D. M.; Storey, J. W.; Thompson, R. I.; van der Werf, D. P.; Wurtele, J. S.; Yamazaki, Y.; Alpha Collaboration

    2008-02-01

    Substantial progress has been made in the last few years in the nascent field of antihydrogen physics. The next big step forward is expected to be the trapping of the formed antihydrogen atoms using a magnetic multipole trap. ALPHA is a new international project that started to take data in 2006 at CERN's Antiproton Decelerator facility. The primary goal of ALPHA is stable trapping of cold antihydrogen atoms to facilitate measurements of its properties. We discuss the status of the ALPHA project and the prospects for antihydrogen trapping.

  13. Towards trapped antihydrogen

    CERN Document Server

    Jorgensen, L V; Bertsche, W; Boston, A; Bowe, P D; Cesar, C L; Chapman, S; Charlton, M; Fajans, J; Fujiwara, M C; Funakoshi, R; Gill, D R; Hangst, J S; Hayano, R S; Hydomako, R; Jenkins, M J; Kurchaninov, L; Madsen, N; Nolan, P; Olchanski, K; Olin, A; Page, R D; Povilus, A; Robicheaux, F; Sarid, E; Silveira, D M; Storey, J W; Thompson, R I; van der Werf, D P; Wurtele, J S; Yamazaki, Y

    2008-01-01

    Substantial progress has been made in the last few years in the nascent field of antihydrogen physics. The next big step forward is expected to be the trapping of the formed antihydrogen atoms using a magnetic multipole trap. ALPHA is a new international project that started to take data in 2006 at CERN’s Antiproton Decelerator facility. The primary goal of ALPHA is stable trapping of cold antihydrogen atoms to facilitate measurements of its properties. We discuss the status of the ALPHA project and the prospects for antihydrogen trapping.

  14. Health safety nets can break cycles of poverty and disease: a stochastic ecological model.

    Science.gov (United States)

    Plucinski, Mateusz M; Ngonghala, Calistus N; Bonds, Matthew H

    2011-12-07

    The persistence of extreme poverty is increasingly attributed to dynamic interactions between biophysical processes and economics, though there remains a dearth of integrated theoretical frameworks that can inform policy. Here, we present a stochastic model of disease-driven poverty traps. Whereas deterministic models can result in poverty traps that can only be broken by substantial external changes to the initial conditions, in the stochastic model there is always some probability that a population will leave or enter a poverty trap. We show that a 'safety net', defined as an externally enforced minimum level of health or economic conditions, can guarantee ultimate escape from a poverty trap, even if the safety net is set within the basin of attraction of the poverty trap, and even if the safety net is only in the form of a public health measure. Whereas the deterministic model implies that small improvements in initial conditions near the poverty-trap equilibrium are futile, the stochastic model suggests that the impact of changes in the location of the safety net on the rate of development may be strongest near the poverty-trap equilibrium.

  15. Search for trapped antihydrogen

    Science.gov (United States)

    Andresen, G. B.; Ashkezari, M. D.; Baquero-Ruiz, M.; Bertsche, W.; Bowe, P. D.; Bray, C. C.; Butler, E.; Cesar, C. L.; Chapman, S.; Charlton, M.; Fajans, J.; Friesen, T.; Fujiwara, M. C.; Gill, D. R.; Hangst, J. S.; Hardy, W. N.; Hayano, R. S.; Hayden, M. E.; Humphries, A. J.; Hydomako, R.; Jonsell, S.; Jørgensen, L. V.; Kurchaninov, L.; Lambo, R.; Madsen, N.; Menary, S.; Nolan, P.; Olchanski, K.; Olin, A.; Povilus, A.; Pusa, P.; Robicheaux, F.; Sarid, E.; Seif El Nasr, S.; Silveira, D. M.; So, C.; Storey, J. W.; Thompson, R. I.; van der Werf, D. P.; Wilding, D.; Wurtele, J. S.; Yamazaki, Y.; Alpha Collaboration

    2011-01-01

    We present the results of an experiment to search for trapped antihydrogen atoms with the ALPHA antihydrogen trap at the CERN Antiproton Decelerator. Sensitive diagnostics of the temperatures, sizes, and densities of the trapped antiproton and positron plasmas have been developed, which in turn permitted development of techniques to precisely and reproducibly control the initial experimental parameters. The use of a position-sensitive annihilation vertex detector, together with the capability of controllably quenching the superconducting magnetic minimum trap, enabled us to carry out a high-sensitivity and low-background search for trapped synthesised antihydrogen atoms. We aim to identify the annihilations of antihydrogen atoms held for at least 130 ms in the trap before being released over ∼30 ms. After a three-week experimental run in 2009 involving mixing of 10 7 antiprotons with 1.3×10 positrons to produce 6×10 antihydrogen atoms, we have identified six antiproton annihilation events that are consistent with the release of trapped antihydrogen. The cosmic ray background, estimated to contribute 0.14 counts, is incompatible with this observation at a significance of 5.6 sigma. Extensive simulations predict that an alternative source of annihilations, the escape of mirror-trapped antiprotons, is highly unlikely, though this possibility has not yet been ruled out experimentally.

  16. Heteroptera attracted to butterfly traps baited with fish or shrimp carrion

    Science.gov (United States)

    Records of Heteroptera collected at butterfly traps baited with fish or shrimp carrion during collecting trips to Bolivia, Brazil, Colombia, Ecuador, and Peru are presented. Traps consisted of a cylinder of net fabric (about 35 cm diam, 75 cm length) attached on the top and bottom to square pieces ...

  17. Trapping radioactive ions

    CERN Document Server

    Kluge, Heinz-Jürgen

    2004-01-01

    Trapping devices for atomic and nuclear physics experiments with radioactive ions are becoming more and more important at accelerator facilities. While about ten years ago only one online Penning trap experiment existed, namely ISOLTRAP at ISOLDE/CERN, meanwhile almost every radioactive beam facility has installed or plans an ion trap setup. This article gives an overview on ion traps in the operation, construction or planing phase which will be used for fundamental studies with short-lived radioactive nuclides such as mass spectrometry, laser spectroscopy and nuclear decay spectroscopy. In addition, this article summarizes the use of gas cells and radiofrequency quadrupole (Paul) traps at different facilities as a versatile tool for ion beam manipulation like retardation, cooling, bunching, and cleaning.

  18. Coloured Petri Nets

    DEFF Research Database (Denmark)

    Jensen, Kurt

    1991-01-01

    use of CP-nets — because it means that the function representation and the translations (which are a bit mathematically complex) no longer are parts of the basic definition of CP-nets. Instead they are parts of the invariant method (which anyway demands considerable mathematical skills...... the different kinds of analysis. It has, however, turned out that it only is necessary to turn to functions when we deal with invariant analysis, and this means that we now use the expression representation for all purposes — except for the calculation of invariants. This change is important for the practical...

  19. Biological Petri Nets

    CERN Document Server

    Wingender, E

    2011-01-01

    It was suggested some years ago that Petri nets might be well suited to modeling metabolic networks, overcoming some of the limitations encountered by the use of systems employing ODEs (ordinary differential equations). Much work has been done since then which confirms this and demonstrates the usefulness of this concept for systems biology. Petri net technology is not only intuitively understood by scientists trained in the life sciences, it also has a robust mathematical foundation and provides the required degree of flexibility. As a result it appears to be a very promising approach to mode

  20. 1985-86 Trapping Proposal

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — This Annual Trapping Plan for the 1985-1986 trapping season at Clarence Cannon NWR outlines rules and regulations for the trapping of beaver and muskrat on the...

  1. Safety nets or straitjackets?

    DEFF Research Database (Denmark)

    Ilsøe, Anna

    2012-01-01

    Does regulation of working hours at national and sector level impose straitjackets, or offer safety nets to employees seeking working time flexibility? This article compares legislation and collective agreements in the metal industries of Denmark, Germany and the USA. The industry has historically...

  2. Nonmetro Net Outmigration Stops.

    Science.gov (United States)

    Cromartie, John B.

    1992-01-01

    Annual population losses from net migration for nonmetro areas declined from 0.38-0.20 percent during the period of 1988-91. However, annual inmigration and outmigration flows were consistently above 1.5 million (about 3 percent of nonmetro population). During the three-year period, nonmetro areas consistently lost young adults and those with…

  3. Net4Care platform

    DEFF Research Database (Denmark)

    2012-01-01

    , that in turn enables general practitioners and clinical staff to view observations. Use the menus above to explore the site's information resources. To get started, follow the short Hello, World! tutorial. The Net4Care project is funded by The Central Denmark Region and EU via Caretech Innovation....

  4. Coloured Petri Nets

    CERN Document Server

    Jensen, Kurt

    2009-01-01

    Coloured Petri Nets (CPN) is a graphical language for modelling and validating concurrent and distributed systems, and other systems in which concurrency plays a major role. This book introduces the constructs of the CPN modelling language and presents the related analysis methods. It provides a comprehensive road map for the practical use of CPN.

  5. Mobile robot sense net

    Science.gov (United States)

    Konolige, Kurt G.; Gutmann, Steffen; Guzzoni, Didier; Ficklin, Robert W.; Nicewarner, Keith E.

    1999-08-01

    Mobile robot hardware and software is developing to the point where interesting applications for groups of such robots can be contemplated. We envision a set of mobots acting to map and perform surveillance or other task within an indoor environment (the Sense Net). A typical application of the Sense Net would be to detect survivors in buildings damaged by earthquake or other disaster, where human searchers would be put a risk. As a team, the Sense Net could reconnoiter a set of buildings faster, more reliably, and more comprehensibly than an individual mobot. The team, for example, could dynamically form subteams to perform task that cannot be done by individual robots, such as measuring the range to a distant object by forming a long baseline stereo sensor form a pari of mobots. In addition, the team could automatically reconfigure itself to handle contingencies such as disabled mobots. This paper is a report of our current progress in developing the Sense Net, after the first year of a two-year project. In our approach, each mobot has sufficient autonomy to perform several tasks, such as mapping unknown areas, navigating to specific positions, and detecting, tracking, characterizing, and classifying human and vehicular activity. We detail how some of these tasks are accomplished, and how the mobot group is tasked.

  6. Net4Care platform

    DEFF Research Database (Denmark)

    2012-01-01

    , that in turn enables general practitioners and clinical staff to view observations. Use the menus above to explore the site's information resources. To get started, follow the short Hello, World! tutorial. The Net4Care project is funded by The Central Denmark Region and EU via Caretech Innovation....

  7. Game Theory .net.

    Science.gov (United States)

    Shor, Mikhael

    2003-01-01

    States making game theory relevant and accessible to students is challenging. Describes the primary goal of GameTheory.net is to provide interactive teaching tools. Indicates the site strives to unite educators from economics, political and computer science, and ecology by providing a repository of lecture notes and tests for courses using…

  8. Nematode-Trapping Fungi.

    Science.gov (United States)

    Jiang, Xiangzhi; Xiang, Meichun; Liu, Xingzhong

    2017-01-01

    Nematode-trapping fungi are a unique and intriguing group of carnivorous microorganisms that can trap and digest nematodes by means of specialized trapping structures. They can develop diverse trapping devices, such as adhesive hyphae, adhesive knobs, adhesive networks, constricting rings, and nonconstricting rings. Nematode-trapping fungi have been found in all regions of the world, from the tropics to Antarctica, from terrestrial to aquatic ecosystems. They play an important ecological role in regulating nematode dynamics in soil. Molecular phylogenetic studies have shown that the majority of nematode-trapping fungi belong to a monophyletic group in the order Orbiliales (Ascomycota). Nematode-trapping fungi serve as an excellent model system for understanding fungal evolution and interaction between fungi and nematodes. With the development of molecular techniques and genome sequencing, their evolutionary origins and divergence, and the mechanisms underlying fungus-nematode interactions have been well studied. In recent decades, an increasing concern about the environmental hazards of using chemical nematicides has led to the application of these biological control agents as a rapidly developing component of crop protection.

  9. Extracellular Matrix Proteins

    Directory of Open Access Journals (Sweden)

    Linda Christian Carrijo-Carvalho

    2012-01-01

    Full Text Available Lipocalin family members have been implicated in development, regeneration, and pathological processes, but their roles are unclear. Interestingly, these proteins are found abundant in the venom of the Lonomia obliqua caterpillar. Lipocalins are β-barrel proteins, which have three conserved motifs in their amino acid sequence. One of these motifs was shown to be a sequence signature involved in cell modulation. The aim of this study is to investigate the effects of a synthetic peptide comprising the lipocalin sequence motif in fibroblasts. This peptide suppressed caspase 3 activity and upregulated Bcl-2 and Ki-67, but did not interfere with GPCR calcium mobilization. Fibroblast responses also involved increased expression of proinflammatory mediators. Increase of extracellular matrix proteins, such as collagen, fibronectin, and tenascin, was observed. Increase in collagen content was also observed in vivo. Results indicate that modulation effects displayed by lipocalins through this sequence motif involve cell survival, extracellular matrix remodeling, and cytokine signaling. Such effects can be related to the lipocalin roles in disease, development, and tissue repair.

  10. Microfabricated Waveguide Atom Traps.

    Energy Technology Data Exchange (ETDEWEB)

    Jau, Yuan-Yu

    2017-09-01

    A nano - scale , microfabricated waveguide structure can in - principle be used to trap atoms in well - defined locations and enable strong photon - atom interactions . A neutral - atom platf orm based on this microfabrication technology will be pre - aligned , which is especially important for quantum - control applications. At present, there is still no reported demonstration of evanescent - field atom trapping using a microfabricated waveguide structure. We described the capabilities established by our team for future development of the waveguide atom - trapping technology at SNL and report our studies to overcome the technical challenges of loading cold atoms into the waveguide atom traps, efficient and broadband optical coupling to a waveguide, and the waveguide material for high - power optical transmission. From the atomic - physics and the waveguide modeling, w e have shown that a square nano - waveguide can be utilized t o achieve better atomic spin squeezing than using a nanofiber for first time.

  11. Induction of trap formation in nematode-trapping fungi by bacteria-released ammonia.

    Science.gov (United States)

    Su, H N; Xu, Y Y; Wang, X; Zhang, K Q; Li, G H

    2016-04-01

    A total of 11 bacterial strains were assayed for bacteria-induced trap formation in the nematode-trapping fungus Arthrobotrys oligospora YMF1·01883 with two-compartmented Petri dish. These strains were identified on the basis of their 16S rRNA gene sequences. Volatile organic compounds (VOCs) of eight isolates were extracted using solid-phase micro-extraction (SPME) and their structures were identified based on gas chromatography-mass spectrometry (GC-MS). At the same time, all isolates were used for quantitative measurement of ammonia by the indophenol blue method. The effects of pure commercial compounds on inducement of trap formation in A. oligospora were tested. Taken together, results demonstrated that the predominant bacterial volatile compound inducing trap formation was ammonia. Meanwhile, ammonia also played a role in other nematode-trapping fungi, including Arthrobotrys guizhouensis YMF1·00014, producing adhesive nets; Dactylellina phymatopaga YMF1·01474, producing adhesive knobs; Dactylellina cionopaga YMF1·01472, producing adhesive columns and Drechslerella brochopaga YMF1·01829, producing constricting rings.

  12. Search For Trapped Antihydrogen

    CERN Document Server

    Andresen, Gorm B; Baquero-Ruiz, Marcelo; Bertsche, William; Bowe, Paul D; Bray, Crystal C; Butler, Eoin; Cesar, Claudio L; Chapman, Steven; Charlton, Michael; Fajans, Joel; Friesen, Tim; Fujiwara, Makoto C; Gill, David R; Hangst, Jeffrey S; Hardy, Walter N; Hayano, Ryugo S; Hayden, Michael E; Humphries, Andrew J; Hydomako, Richard; Jonsell, Svante; Jørgensen, Lars V; Kurchaninov, Lenoid; Lambo, Ricardo; Madsen, Niels; Menary, Scott; Nolan, Paul; Olchanski, Konstantin; Olin, Art; Povilus, Alexander; Pusa, Petteri; Robicheaux, Francis; Sarid, Eli; Nasr, Sarah Seif El; Silveira, Daniel M; So, Chukman; Storey, James W; Thompson, Robert I; van der Werf, Dirk P; Wilding, Dean; Wurtele, Jonathan S; Yamazaki, Yasunori

    2011-01-01

    We present the results of an experiment to search for trapped antihydrogen atoms with the ALPHA antihydrogen trap at the CERN Antiproton Decelerator. Sensitive diagnostics of the temperatures, sizes, and densities of the trapped antiproton and positron plasmas have been developed, which in turn permitted development of techniques to precisely and reproducibly control the initial experimental parameters. The use of a position-sensitive annihilation vertex detector, together with the capability of controllably quenching the superconducting magnetic minimum trap, enabled us to carry out a high-sensitivity and low-background search for trapped synthesised antihydrogen atoms. We aim to identify the annihilations of antihydrogen atoms held for at least 130 ms in the trap before being released over ~30 ms. After a three-week experimental run in 2009 involving mixing of 10^7 antiprotons with 1.3 10^9 positrons to produce 6 10^5 antihydrogen atoms, we have identified six antiproton annihilation events that are consist...

  13. Microfabricated cylindrical ion trap

    Science.gov (United States)

    Blain, Matthew G.

    2005-03-22

    A microscale cylindrical ion trap, having an inner radius of order one micron, can be fabricated using surface micromachining techniques and materials known to the integrated circuits manufacturing and microelectromechanical systems industries. Micromachining methods enable batch fabrication, reduced manufacturing costs, dimensional and positional precision, and monolithic integration of massive arrays of ion traps with microscale ion generation and detection devices. Massive arraying enables the microscale cylindrical ion trap to retain the resolution, sensitivity, and mass range advantages necessary for high chemical selectivity. The microscale CIT has a reduced ion mean free path, allowing operation at higher pressures with less expensive and less bulky vacuum pumping system, and with lower battery power than conventional- and miniature-sized ion traps. The reduced electrode voltage enables integration of the microscale cylindrical ion trap with on-chip integrated circuit-based rf operation and detection electronics (i.e., cell phone electronics). Therefore, the full performance advantages of microscale cylindrical ion traps can be realized in truly field portable, handheld microanalysis systems.

  14. Extracellular matrix structure.

    Science.gov (United States)

    Theocharis, Achilleas D; Skandalis, Spyros S; Gialeli, Chrysostomi; Karamanos, Nikos K

    2016-02-01

    Extracellular matrix (ECM) is a non-cellular three-dimensional macromolecular network composed of collagens, proteoglycans/glycosaminoglycans, elastin, fibronectin, laminins, and several other glycoproteins. Matrix components bind each other as well as cell adhesion receptors forming a complex network into which cells reside in all tissues and organs. Cell surface receptors transduce signals into cells from ECM, which regulate diverse cellular functions, such as survival, growth, migration, and differentiation, and are vital for maintaining normal homeostasis. ECM is a highly dynamic structural network that continuously undergoes remodeling mediated by several matrix-degrading enzymes during normal and pathological conditions. Deregulation of ECM composition and structure is associated with the development and progression of several pathologic conditions. This article emphasizes in the complex ECM structure as to provide a better understanding of its dynamic structural and functional multipotency. Where relevant, the implication of the various families of ECM macromolecules in health and disease is also presented.

  15. Army Net Zero Prove Out. Net Zero Waster Best Practices

    Science.gov (United States)

    2014-11-18

    Colorado • Fort Bliss, Texas These sites served as test beds for the Army’s Net Zero Initiative, specifically, Net Zero Water , and the Army provided...have sustainability, energy efficiency, water conservation, recycling, pollution prevention, and green procurement programs in place that they can...ARMY NET ZERO PROVE OUT Final Net Zero Water Best Practices November 18, 2014 Distribution A Approved for public release

  16. Microbially enhanced carbon capture and storage by mineral-trapping and solubility-trapping.

    Science.gov (United States)

    Mitchell, Andrew C; Dideriksen, Knud; Spangler, Lee H; Cunningham, Alfred B; Gerlach, Robin

    2010-07-01

    The potential of microorganisms for enhancing carbon capture and storage (CCS) via mineral-trapping (where dissolved CO(2) is precipitated in carbonate minerals) and solubility trapping (as dissolved carbonate species in solution) was investigated. The bacterial hydrolysis of urea (ureolysis) was investigated in microcosms including synthetic brine (SB) mimicking a prospective deep subsurface CCS site with variable headspace pressures [p(CO(2))] of (13)C-CO(2). Dissolved Ca(2+) in the SB was completely precipitated as calcite during microbially induced hydrolysis of 5-20 g L(-1) urea. The incorporation of carbonate ions from (13)C-CO(2) ((13)C-CO(3)(2-)) into calcite increased with increasing p((13)CO(2)) and increasing urea concentrations: from 8.3% of total carbon in CaCO(3) at 1 g L(-1) to 31% at 5 g L(-1), and 37% at 20 g L(-1). This demonstrated that ureolysis was effective at precipitating initially gaseous [CO(2)(g)] originating from the headspace over the brine. Modeling the change in brine chemistry and carbonate precipitation after equilibration with the initial p(CO(2)) demonstrated that no net precipitation of CO(2)(g) via mineral-trapping occurred, since urea hydrolysis results in the production of dissolved inorganic carbon. However, the pH increase induced by bacterial ureolysis generated a net flux of CO(2)(g) into the brine. This reduced the headspace concentration of CO(2) by up to 32 mM per 100 mM urea hydrolyzed because the capacity of the brine for carbonate ions was increased, thus enhancing the solubility-trapping capacity of the brine. Together with the previously demonstrated permeability reduction of rock cores at high pressure by microbial biofilms and resilience of biofilms to supercritical CO(2), this suggests that engineered biomineralizing biofilms may enhance CCS via solubility-trapping, mineral formation, and CO(2)(g) leakage reduction.

  17. Net one, net two: the primary care network income statement.

    Science.gov (United States)

    Halley, M D; Little, A W

    1999-10-01

    Although hospital-owned primary care practices have been unprofitable for most hospitals, some hospitals are achieving competitive advantage and sustainable practice operations. A key to the success of some has been a net income reporting tool that separates practice operating expenses from the costs of creating and operating a network of practices to help healthcare organization managers, physicians, and staff to identify opportunities to improve the network's financial performance. This "Net One, Net Two" reporting allows operations leadership to be held accountable for Net One expenses and strategic leadership to be held accountable for Net Two expenses.

  18. Proof nets for lingusitic analysis

    NARCIS (Netherlands)

    Moot, R.C.A.

    2002-01-01

    This book investigates the possible linguistic applications of proof nets, redundancy free representations of proofs, which were introduced by Girard for linear logic. We will adapt the notion of proof net to allow the formulation of a proof net calculus which is soundand complete for the multimoda

  19. Boom Booom Net Radio

    DEFF Research Database (Denmark)

    Grimshaw, Mark Nicholas; Yong, Louisa; Dobie, Ian

    1999-01-01

    of an existing Internet radio station; Boom Booom Net Radio. Whilst necessity dictates some use of technology-related terminology, wherever possible we have endeavoured to keep such jargon to a minimum and to either explain it in the text or to provide further explanation in the appended glossary.......Internet radio is one of the growth areas of the Internet but, as this article will show, is fraught with difficulties and frustration for both the modestly-funded broadcaster (bitcaster) and the listener. The article will illustrate some of these problems by means of a short case study...

  20. Safety nets or straitjackets?

    DEFF Research Database (Denmark)

    Ilsøe, Anna

    2012-01-01

    Does regulation of working hours at national and sector level impose straitjackets, or offer safety nets to employees seeking working time flexibility? This article compares legislation and collective agreements in the metal industries of Denmark, Germany and the USA. The industry has historically...... been trend-setting for collective bargaining in all three countries, but with very different effects on working time. Organized decentralization seems to pave the way for fewer straitjackets, whereas the opposite seems to be the case with regard to disorganized decentralization....

  1. Master Robotic Net

    Directory of Open Access Journals (Sweden)

    Vladimir Lipunov

    2010-01-01

    Full Text Available The main goal of the MASTER-Net project is to produce a unique fast sky survey with all sky observed over a single night down to a limiting magnitude of 19-20. Such a survey will make it possible to address a number of fundamental problems: search for dark energy via the discovery and photometry of supernovae (including SNIa, search for exoplanets, microlensing effects, discovery of minor bodies in the Solar System, and space-junk monitoring. All MASTER telescopes can be guided by alerts, and we plan to observe prompt optical emission from gamma-ray bursts synchronously in several filters and in several polarization planes.

  2. High-level Petri Nets

    DEFF Research Database (Denmark)

    to their work can be found in the individual papers and in the available bibliographies of Petri nets, e.g., Stefan Dress et. al: Bibliography of Petri Nets. In: G. Rozenberg (ed.): Advances in Petri Nets 1987, Lecture Notes in Computer Science, vol. 266, Springer-Verlag 1987, 309-451. Updated versions...... of this bibliography will appear in Advances of Petri Nets with regular intervals. In the preparation of the book the editors have been assisted by an advisory board consisting of the following well-known Petri net researchers: M. Ajmone-Marsan, H.J. Genrich, C. Girault, W. Reisig, M. Silva and P.S. Thiagarajan...

  3. Penning trap at IGISOL

    Energy Technology Data Exchange (ETDEWEB)

    Szerypo, J. E-mail: jerzy.szerypo@phys.jyu.fi; Jokinen, A.; Kolhinen, V.S.; Nieminen, A.; Rinta-Antila, S.; Aeystoe, J

    2002-04-22

    The IGISOL facility at the Department of Physics of the University of Jyvaeskylae (JYFL) is delivering radioactive beams of short-lived exotic nuclei, in particular the neutron-rich isotopes from the fission reaction. These nuclei are studied with the nuclear spectroscopy methods. In order to substantially increase the quality and sensitivity of such studies, the beam should undergo beam handling: cooling, bunching and isobaric purification. The first two processes are performed with the use of an RFQ cooler/buncher. The isobaric purification will be made by a Penning trap placed after the RF-cooler element. This contribution describes the current status of the Penning trap project and its future prospects. The latter comprise the precise nuclear mass measurements, nuclear spectroscopy in the Penning trap interior as well as the laser spectroscopy on the extracted beams.

  4. NETS FOR PEACH PROTECTED CULTIVATION

    Directory of Open Access Journals (Sweden)

    Evelia Schettini

    2012-06-01

    Full Text Available The aim of this paper was to investigate the radiometric properties of coloured nets used to protect a peach cultivation. The modifications of the solar spectral distribution, mainly in the R and FR wavelength band, influence plant photomorphogenesis by means of the phytochrome and cryptochrome. The phytochrome response is characterized in terms of radiation rate in the red wavelengths (R, 600-700 nm to that in the farred radiation (FR, 700-800 nm, i.e. the R/FR ratio. The effects of the blue radiation (B, 400-500 nm is investigated by the ratio between the blue radiation and the far-red radiation, i.e. the B/FR ratio. A BLUE net, a RED net, a YELLOW net, a PEARL net, a GREY net and a NEUTRAL net were tested in Bari (Italy, latitude 41° 05’ N. Peach trees were located in pots inside the greenhouses and in open field. The growth of the trees cultivated in open field was lower in comparison to the growth of the trees grown under the nets. The RED, PEARL, YELLOW and GREY nets increased the growth of the trees more than the other nets. The nets positively influenced the fruit characteristics, such as fruit weight and flesh firmness.

  5. Trapping molecules on chips

    CERN Document Server

    Santambrogio, Gabriele

    2015-01-01

    In the last years, it was demonstrated that neutral molecules can be loaded on a microchip directly from a supersonic beam. The molecules are confined in microscopic traps that can be moved smoothly over the surface of the chip. Once the molecules are trapped, they can be decelerated to a standstill, for instance, or pumped into selected quantum states by laser light or microwaves. Molecules are detected on the chip by time-resolved spatial imaging, which allows for the study of the distribution in the phase space of the molecular ensemble.

  6. Redesigning octopus traps

    Directory of Open Access Journals (Sweden)

    Eduarda Gomes

    2014-06-01

    In order to minimise the identified problems in the actual traps, the present work proposes a new design with the aim of reducing the volume and weight during transport, and also during onshore storage. Alternative materials to avoid corrosion and formation of encrustations were also proposed.

  7. Art/Net/Work

    DEFF Research Database (Denmark)

    Andersen, Christian Ulrik; Lindstrøm, Hanne

    2006-01-01

    The seminar Art|Net|Work deals with two important changes in our culture. On one side, the network has become essential in the latest technological development. The Internet has entered a new phase, Web 2.0, including the occurrence of as ‘Wiki’s’, ‘Peer-2-Peer’ distribution, user controlled...... taxonomies (‘Folksonomy’) and ‘Weblogs’. Also, platforms, programming and software are today very often created in open communities – as seen in the ‘Free/Open Source’ movement. On the other side, following the technological development, the network also has become essential in the art sphere. Artists focus...... on the ‘network’ itself as a phenomenon and are often using technological networks as a mean of production and distribution. This changes the artistic practice and the distribution channels of art works – and the traditional notions of ‘work’, ‘origin’ and ‘rights’ are increasingly perceived as limiting...

  8. Boom Booom Net Radio

    DEFF Research Database (Denmark)

    Grimshaw, Mark Nicholas; Yong, Louisa; Dobie, Ian

    1999-01-01

    Internet radio is one of the growth areas of the Internet but, as this article will show, is fraught with difficulties and frustration for both the modestly-funded broadcaster (bitcaster) and the listener. The article will illustrate some of these problems by means of a short case study of an exi......Internet radio is one of the growth areas of the Internet but, as this article will show, is fraught with difficulties and frustration for both the modestly-funded broadcaster (bitcaster) and the listener. The article will illustrate some of these problems by means of a short case study...... of an existing Internet radio station; Boom Booom Net Radio. Whilst necessity dictates some use of technology-related terminology, wherever possible we have endeavoured to keep such jargon to a minimum and to either explain it in the text or to provide further explanation in the appended glossary....

  9. Extracellular vesicles for drug delivery

    NARCIS (Netherlands)

    Vader, Pieter; Mol, Emma A; Pasterkamp, Gerard; Schiffelers, Raymond M

    Extracellular vesicles (EVs) are cell-derived membrane vesicles, and represent an endogenous mechanism for intercellular communication. Since the discovery that EVs are capable of functionally transferring biological information, the potential use of EVs as drug delivery vehicles has gained

  10. NetView technical research

    Science.gov (United States)

    1993-01-01

    This is the Final Technical Report for the NetView Technical Research task. This report is prepared in accordance with Contract Data Requirements List (CDRL) item A002. NetView assistance was provided and details are presented under the following headings: NetView Management Systems (NMS) project tasks; WBAFB IBM 3090; WPAFB AMDAHL; WPAFB IBM 3084; Hill AFB; McClellan AFB AMDAHL; McClellan AFB IBM 3090; and Warner-Robins AFB.

  11. Optical investigation of intra- and extra-cellular dynamics

    DEFF Research Database (Denmark)

    Leijnse, Natascha

    In this thesis we investigated intra- and extra-cellular dynamics with the help of optical tweezers and confocal imaging. Firstly, we measured in vivo diffusion inside endothelial cells by tracking lipid granules and compared the results to in vitro diffusion of polystyrene beads in wormlike...... micelles. Furthermore, we studied F-actin dynamics in, and curvature sensing of, a receptor and its ligand on artificially pulled membrane tethers. Force measurements between a cell and an approaching optically trapped bead were conducted in order to probe the endothelial cell's glycocalyx. Finally...

  12. Optical investigation of intra- and extra-cellular dynamics

    DEFF Research Database (Denmark)

    Leijnse, Natascha

    In this thesis we investigated intra- and extra-cellular dynamics with the help of optical tweezers and confocal imaging. Firstly, we measured in vivo diffusion inside endothelial cells by tracking lipid granules and compared the results to in vitro diffusion of polystyrene beads in wormlike...... micelles. Furthermore, we studied F-actin dynamics in, and curvature sensing of, a receptor and its ligand on artificially pulled membrane tethers. Force measurements between a cell and an approaching optically trapped bead were conducted in order to probe the endothelial cell's glycocalyx. Finally...

  13. ASP.NET MVC 2

    CERN Document Server

    Chiaretta, Simone

    2010-01-01

    This Wrox Blox is for .NET developers who already have experiencedeveloping web applications with ASP.NET MVC and want to learn thenew features introduced in the new release of ASP.NET MVC. ASP.NETMVC 2 has been built with the goal to increase productivity andmake ASP.NET MVC ready for enterprise development. This Wrox Bloxguides the reader through the new features by explaining how to usedata scaffolding and automatic validation, how to write customvalidators, how to create multi-area projects, and how and when touse async controllers. It also covers the new automatic HTMLEncoding feature int

  14. Characterizing optical dipole trap via fluorescence of trapped cesium atoms

    Institute of Scientific and Technical Information of China (English)

    LIU Tao; GENG Tao; YAN Shubin; LI Gang; ZHANG Jing; WANG Junmin; PENG Kunchi; ZHANG Tiancai

    2006-01-01

    Optical dipole trap (ODT) is becoming an important tool of manipulating neutral atoms. In this paper ODT is realized with a far-off resonant laser beam strongly focused in the magneto-optical trap (MOT) of cesium atoms. The light shift is measured by simply monitoring the fluorescence of the atoms in the magneto-optical trap and the optical dipole trap simultaneously. The advantages of our experimental scheme are discussed, and the effect of the beam waist and power on the potential of dipole trap as well as heating rate is analyzed.

  15. Traps for neutral radioactive atoms

    CERN Document Server

    Sprouse, G D; Grossman, J S; Orozco, L A; Pearson, M R

    2002-01-01

    We describe several methods for efficiently injecting a small number of radioactive atoms into a laser trap. The characteristics of laser traps that make them desirable for physics experiments are discussed and several different experimental directions are described. We describe recent experiments with the alkali element Fr and point to future directions of the neutral atom trapping program.

  16. Receptor-like activity evoked by extracellular ADP in Arabidopsis root epidermal plasma membrane.

    Science.gov (United States)

    Demidchik, Vadim; Shang, Zhonglin; Shin, Ryoung; Colaço, Renato; Laohavisit, Anuphon; Shabala, Sergey; Davies, Julia M

    2011-07-01

    Extracellular purine nucleotides are implicated in the control of plant development and stress responses. While extracellular ATP is known to activate transcriptional pathways via plasma membrane (PM) NADPH oxidase and calcium channel activation, very little is known about signal transduction by extracellular ADP. Here, extracellular ADP was found to activate net Ca(2+) influx in roots of Arabidopsis (Arabidopsis thaliana) and transiently elevate cytosolic free Ca(2+) in root epidermal protoplasts. An inward Ca(2+)-permeable conductance in root epidermal PM was activated within 1 s of ADP application and repeated application evoked a smaller current. Such response speed and densitization are consistent with operation of equivalents to animal ionotropic purine receptors, although to date no equivalent genes for such receptors have been identified in higher plants. In contrast to ATP, extracellular ADP did not evoke accumulation of intracellular reactive oxygen species. While high concentrations of ATP caused net Ca(2+) efflux from roots, equivalent concentrations of ADP caused net influx. Overall the results point to a discrete ADP signaling pathway, reliant on receptor-like activity at the PM.

  17. Receptor-Like Activity Evoked by Extracellular ADP in Arabidopsis Root Epidermal Plasma Membrane1

    Science.gov (United States)

    Demidchik, Vadim; Shang, Zhonglin; Shin, Ryoung; Colaço, Renato; Laohavisit, Anuphon; Shabala, Sergey; Davies, Julia M.

    2011-01-01

    Extracellular purine nucleotides are implicated in the control of plant development and stress responses. While extracellular ATP is known to activate transcriptional pathways via plasma membrane (PM) NADPH oxidase and calcium channel activation, very little is known about signal transduction by extracellular ADP. Here, extracellular ADP was found to activate net Ca2+ influx in roots of Arabidopsis (Arabidopsis thaliana) and transiently elevate cytosolic free Ca2+ in root epidermal protoplasts. An inward Ca2+-permeable conductance in root epidermal PM was activated within 1 s of ADP application and repeated application evoked a smaller current. Such response speed and densitization are consistent with operation of equivalents to animal ionotropic purine receptors, although to date no equivalent genes for such receptors have been identified in higher plants. In contrast to ATP, extracellular ADP did not evoke accumulation of intracellular reactive oxygen species. While high concentrations of ATP caused net Ca2+ efflux from roots, equivalent concentrations of ADP caused net influx. Overall the results point to a discrete ADP signaling pathway, reliant on receptor-like activity at the PM. PMID:21562328

  18. Detection of Trapped Antihydrogen

    CERN Document Server

    Hydomako, Richard Allan

    The ALPHA experiment is an international effort to produce, trap, and perform precision spectroscopic measurements on antihydrogen (the bound state of a positron and an antiproton). Based at the Antiproton Decelerator (AD) facility at CERN, the ALPHA experiment has recently magnetically confined antihydrogen atoms for the first time. A crucial element in the observation of trapped antihydrogen is ALPHA’s silicon vertexing detector. This detector contains sixty silicon modules arranged in three concentric layers, and is able to determine the three-dimensional location of the annihilation of an antihydrogen atom by reconstructing the trajectories of the produced annihilation products. This dissertation focuses mainly on the methods used to reconstruct the annihilation location. Specifically, the software algorithms used to identify and extrapolate charged particle tracks are presented along with the routines used to estimate the annihilation location from the convergence of the identified tracks. It is shown...

  19. Coherence in Microchip Traps

    CERN Document Server

    Treutlein, P; Steinmetz, T; Hänsch, T W; Reichel, J; Treutlein, Philipp; Hommelhoff, Peter; Steinmetz, Tilo; H\\"ansch, Theodor W.; Reichel, Jakob

    2003-01-01

    We report the coherent manipulation of internal states of neutral atoms in a magnetic microchip trap. Coherence lifetimes exceeding 1 s are observed with atoms at distances of $4-130 \\mu$m from the microchip surface. The coherence lifetime in the microtrap is independent of atom-surface distance and agrees well with the results of similar measurements in macroscopic magnetic traps. Due to the absence of surface-induced decoherence, a miniaturized atomic clock with a relative stability in the $10^{-13}$ range can be realized. For applications in quantum information processing, we propose to use microwave near-fields in the proximity of chip wires to create potentials that depend on the internal state of the atoms.

  20. Ion Trap Quantum Computing

    Science.gov (United States)

    2011-12-01

    an inspiring speech at the MIT Physics of Computation 1st Conference in 1981, Feynman proposed the development of a computer that would obey the...on ion trap based 36 quantum computing for physics and computer science students would include lecture notes, slides, lesson plans, a syllabus...reading lists, videos, demonstrations, and laboratories. 37 LIST OF REFERENCES [1] R. P. Feynman , “Simulating physics with computers,” Int. J

  1. Water-Trapped Worlds

    CERN Document Server

    Menou, Kristen

    2013-01-01

    Although tidally-locked habitable planets orbiting nearby M-dwarf stars are among the best astronomical targets to search for extrasolar life, they may also be deficient in volatiles and water. Climate models for this class of planets show atmospheric transport of water from the dayside to the nightside, where it is precipitated as snow and trapped as ice. Since ice only slowly flows back to the dayside upon accumulation, the resulting hydrological cycle can trap a large amount of water in the form of nightside ice. Using ice sheet dynamical and thermodynamical constraints, I illustrate how planets with less than about a quarter the Earth's oceans could trap most of their surface water on the nightside. This would leave their dayside, where habitable conditions are met, potentially dry. The amount and distribution of residual liquid water on the dayside depend on a variety of geophysical factors, including the efficiency of rock weathering at regulating atmospheric CO2 as dayside ocean basins dry-up. Water-tr...

  2. DMAH ozone measurement net

    Directory of Open Access Journals (Sweden)

    D. Pagès

    2006-01-01

    Full Text Available The complexity of the study of tropospheric ozone lies in the fact that it is a secondary pollutant. It is not emitted by a source, instead its concentration in the air depends on other compounds (especially the nitrogen oxides emitted by motor vehicles and the volatile organic compounds emitted by the industry and the vegetation and meteorological factors (especially solar radiation and temperature. The European legislation compells to make measurements of the tropospheric ozone due to its effects on people (fatigue, irritation of the mucous membranes, aggravation of asthma ... and on environment (decrease of the production of cereals, synergy with plagues .... The measuring net in Catalonia belongs to the Department of Environment and Housing (DMAH. It has a pyramidal structure and it allows a surveillance to notify in case of exceeding a certain threshold. From the registered data of last years it is shown that the number of incidences is related to meteorology. They are more frequent during afternoon and the behaviour of this pollutant is different according to the proximity of the point of measurement to the sources of its precursors.

  3. Net4Care Ecosystem Website

    DEFF Research Database (Denmark)

    Christensen, Henrik Bærbak; Hansen, Klaus Marius; Rasmussen, Morten

    2012-01-01

    The Net4Care platform provides the ability to quickly create quality tele-health applications interfacing with HL7 and IHE/XDS. Net4Care features a modular server- and client-side based on OSGi and aims at providing support for a telemedicine software ecosystem. A typical deployment scenario is a...

  4. Linear Logic on Petri Nets

    DEFF Research Database (Denmark)

    Engberg, Uffe Henrik; Winskel, Glynn

    This article shows how individual Petri nets form models of Girard's intuitionistic linear logic. It explores questions of expressiveness and completeness of linear logic with respect to this interpretation. An aim is to use Petri nets to give an understanding of linear logic and give some apprai...

  5. A Small Universal Petri Net

    Directory of Open Access Journals (Sweden)

    Dmitry A. Zaitsev

    2013-09-01

    Full Text Available A universal deterministic inhibitor Petri net with 14 places, 29 transitions and 138 arcs was constructed via simulation of Neary and Woods' weakly universal Turing machine with 2 states and 4 symbols; the total time complexity is exponential in the running time of their weak machine. To simulate the blank words of the weakly universal Turing machine, a couple of dedicated transitions insert their codes when reaching edges of the working zone. To complete a chain of a given Petri net encoding to be executed by the universal Petri net, a translation of a bi-tag system into a Turing machine was constructed. The constructed Petri net is universal in the standard sense; a weaker form of universality for Petri nets was not introduced in this work.

  6. Relevance of extracellular DNA in rhizosphere

    Science.gov (United States)

    Pietramellara, Giacomo; Ascher, Judith; Baraniya, Divyashri; Arfaioli, Paola; Ceccherini, Maria Teresa; Hawes, Martha

    2013-04-01

    One of the most promising areas for future development is the manipulation of the rhizosphere to produce sustainable and efficient agriculture production systems. Using Omics approaches, to define the distinctive features of eDNA systems and structures, will facilitate progress in rhizo-enforcement and biocontrol studies. The relevance of these studies results clear when we consider the plethora of ecological functions in which eDNA is involved. This fraction can be actively extruded by living cells or discharged during cellular lysis and may exert a key role in the stability and variability of the soil bacterial genome, resulting also a source of nitrogen and phosphorus for plants due to the root's capacity to directly uptake short DNA fragments. The adhesive properties of the DNA molecule confer to eDNA the capacity to inhibit or kill pathogenic bacteria by cation limitation induction, and to facilitate formation of biofilm and extracellular traps (ETs), that may protect microorganisms inhabiting biofilm and plant roots against pathogens and allelopathic substances. The ETs are actively extruded by root border cells when they are dispersed in the rhizosphere, conferring to plants the capacity to extend an endogenous pathogen defence system outside the organism. Moreover, eDNA could be involved in rhizoremediation in heavy metal polluted soil acting as a bioflotation reagent.

  7. Modelling Inteligent Systems with Level Petri Nets

    Directory of Open Access Journals (Sweden)

    Corina BOCĂNEALĂ

    2008-12-01

    Full Text Available Level Petri Nets are formalism for modeling hierarchical multi-agent system. They are a Petri nets extension, allowing tokens to be nets themselves. This paper is inspired by two classes of level Petri nets: object Petri nets and nested Petri nets. We present some concepts from the artificial intelligence field and we use them to illustrate the modeling power of Petri nets with multi levels.

  8. C-terminus of TRAP in Staphylococcus can enhance the activity of lysozyme and lysostaphin

    Institute of Scientific and Technical Information of China (English)

    Guang Yang; Ningsheng Shao; Yaping Gao; Jiannan Feng; Yong Huang; Shaohua Li; Yu Liu; Chuan Liu; Ming Fan; Beifen Shen

    2008-01-01

    In Staphylococcus aureus, the target of RNAⅢ activating protein (TRAP) is a membrane-associated protein whose Cterminus can be used as a vaccine to provide protection against staphylococcal infection. Here, we show for the first time by surface plasmon resonance and enzyme-linked immunosorbent assay that TRAP can specifically bind lysozyme and iysostaphin through its C-terminus (amino acids 155-167) and enhance lysozomal activities in vitro. It was also found that the traP mutant strain is more resistant to iysostaphin than wild-type. Our previous data showed that the C-terminus of TRAP might be extracellular. So our results suggested that the C-terminus of TRAP could act as the specific targeting protein of the lysozyme/lysostaphin on the S. aureus cell wall and the biological significance of the interaction might be to facilitate lysozyme/lysostaphin-mediated cell iysis.

  9. Are mist nets a menace for threatened insects? A first attempt to gather quantitative data

    DEFF Research Database (Denmark)

    Carpaneto, Giuseppe Maria; Mazziotta, Adriano; Maltzeff, Paolo

    2011-01-01

    We attempt to identify the threat to flying beetles imposed by mist nets deployed for bird studies in an area of Mediterranean mosaic of evergreen scrub and deciduous forest in Italy. The present paper is the first attempt to identify the non-target species which die in these artificial traps...

  10. Atomic Coherent Trapping and Properties of Trapped Atom

    Institute of Scientific and Technical Information of China (English)

    YANG Guo-Jian; XIA Li-Xin; XIE Min

    2006-01-01

    Based on the theory of velocity-selective coherent population trapping, we investigate an atom-laser system where a pair of counterpropagating laser fields interact with a three-level atom. The influence of the parametric condition on the properties of the system such as velocity at which the atom is selected to be trapped, time needed for finishing the coherent trapping process, and possible electromagnetically induced transparency of an altrocold atomic medium,etc., is studied.

  11. Cryogenic resonator design for trapped ion experiments in Paul traps

    CERN Document Server

    Brandl, Matthias F; Monz, Thomas; Blatt, Rainer

    2016-01-01

    Trapping ions in Paul traps requires high radio-frequency voltages, which are generated using resonators. When operating traps in a cryogenic environment, an in-vacuum resonator showing low loss is crucial to limit the thermal load to the cryostat. In this study, we present a guide for the design and production of compact, shielded cryogenic resonators. We produced and characterized three different types of resonators and furthermore demonstrate efficient impedance matching of these resonators at cryogenic temperatures.

  12. Nonresonance adiabatic photon trap

    CERN Document Server

    Popov, S S; Burdakov, A V; Ushkova, M Yu

    2016-01-01

    Concept of high efficiency photon storage based on adiabatic confinement between concave mirrors is presented and experimentally investigated. The approach is insensitive to typical for Fabri-Perot cells requirements on quality of accumulated radiation, tolerance of resonator elements and their stability. Experiments have been carried out with the trap, which consists from opposed concave cylindrical mirrors and conjugated with them spherical mirrors. In result, high efficiency for accumulation of radiation with large angular spread and spectrum width has been confirmed. As radiation source a commercial fiber laser has been used.

  13. Atom trap trace analysis

    Energy Technology Data Exchange (ETDEWEB)

    Lu, Z.-T.; Bailey, K.; Chen, C.-Y.; Du, X.; Li, Y.-M.; O' Connor, T. P.; Young, L.

    2000-05-25

    A new method of ultrasensitive trace-isotope analysis has been developed based upon the technique of laser manipulation of neutral atoms. It has been used to count individual {sup 85}Kr and {sup 81}Kr atoms present in a natural krypton sample with isotopic abundances in the range of 10{sup {minus}11} and 10{sup {minus}13}, respectively. The atom counts are free of contamination from other isotopes, elements,or molecules. The method is applicable to other trace-isotopes that can be efficiently captured with a magneto-optical trap, and has a broad range of potential applications.

  14. Conformal Nets II: Conformal Blocks

    Science.gov (United States)

    Bartels, Arthur; Douglas, Christopher L.; Henriques, André

    2017-03-01

    Conformal nets provide a mathematical formalism for conformal field theory. Associated to a conformal net with finite index, we give a construction of the `bundle of conformal blocks', a representation of the mapping class groupoid of closed topological surfaces into the category of finite-dimensional projective Hilbert spaces. We also construct infinite-dimensional spaces of conformal blocks for topological surfaces with smooth boundary. We prove that the conformal blocks satisfy a factorization formula for gluing surfaces along circles, and an analogous formula for gluing surfaces along intervals. We use this interval factorization property to give a new proof of the modularity of the category of representations of a conformal net.

  15. Pro asynchronous programming with .NET

    CERN Document Server

    Blewett, Richard; Ltd, Rock Solid Knowledge

    2014-01-01

    Pro Asynchronous Programming with .NET teaches the essential skill of asynchronous programming in .NET. It answers critical questions in .NET application development, such as: how do I keep my program responding at all times to keep my users happy how do I make the most of the available hardware how can I improve performanceIn the modern world, users expect more and more from their applications and devices, and multi-core hardware has the potential to provide it. But it takes carefully crafted code to turn that potential into responsive, scalable applications.With Pro Asynchronous Programming

  16. Conformal Nets II: Conformal Blocks

    Science.gov (United States)

    Bartels, Arthur; Douglas, Christopher L.; Henriques, André

    2017-08-01

    Conformal nets provide a mathematical formalism for conformal field theory. Associated to a conformal net with finite index, we give a construction of the `bundle of conformal blocks', a representation of the mapping class groupoid of closed topological surfaces into the category of finite-dimensional projective Hilbert spaces. We also construct infinite-dimensional spaces of conformal blocks for topological surfaces with smooth boundary. We prove that the conformal blocks satisfy a factorization formula for gluing surfaces along circles, and an analogous formula for gluing surfaces along intervals. We use this interval factorization property to give a new proof of the modularity of the category of representations of a conformal net.

  17. Petri Nets and Semantics of System Descriptions

    DEFF Research Database (Denmark)

    Jensen, Kurt; Kyng, Morten

    1982-01-01

    This paper discusses the use of Petri nets as a semantic tool in the design of languages and in the construction and analysis of system descriptions. The topics treated are: Languages based on nets. The problem of time in nets. Nets and related models. Nets and formal semantics. Parallel program...

  18. Nanofriction in cold ion traps.

    Science.gov (United States)

    Benassi, A; Vanossi, A; Tosatti, E

    2011-01-01

    Sliding friction between crystal lattices and the physics of cold ion traps are so far non-overlapping fields. Two sliding lattices may either stick and show static friction or slip with dynamic friction; cold ions are known to form static chains, helices or clusters, depending on the trapping conditions. Here we show, based on simulations, that much could be learnt about friction by sliding, through, for example, an electric field, the trapped ion chains over a corrugated potential. Unlike infinite chains, in which the theoretically predicted Aubry transition to free sliding may take place, trapped chains are always pinned. Yet, a properly defined static friction still vanishes Aubry-like at a symmetric-asymmetric structural transition, found for decreasing corrugation in both straight and zig-zag trapped chains. Dynamic friction is also accessible in ringdown oscillations of the ion trap. Long theorized static and dynamic one-dimensional friction phenomena could thus become accessible in future cold ion tribology.

  19. Extracellular vesicles for drug delivery

    NARCIS (Netherlands)

    Vader, Pieter; Mol, Emma A; Pasterkamp, Gerard; Schiffelers, Raymond M

    2016-01-01

    Extracellular vesicles (EVs) are cell-derived membrane vesicles, and represent an endogenous mechanism for intercellular communication. Since the discovery that EVs are capable of functionally transferring biological information, the potential use of EVs as drug delivery vehicles has gained consider

  20. Sweeping beauty: is grassland arthropod community composition effectively estimated by sweep netting?

    Science.gov (United States)

    Spafford, Ryan D; Lortie, Christopher J

    2013-01-01

    Arthropods are critical ecosystem components due to their high diversity and sensitivity to perturbation. Furthermore, due to their ease of capture they are often the focus of environmental health surveys. There is much debate regarding the best sampling method to use in these surveys. Sweep netting and pan trapping are two sampling methods commonly used in agricultural arthropod surveys, but have not been contrasted in natural grassland systems at the community level. The purpose of this study was to determine whether sweep netting was effective at estimating arthropod diversity at the community level in grasslands or if supplemental pan trapping was needed. Arthropods were collected from grassland sites in Montana, USA, in the summer of 2011. The following three standardized evaluation criteria (consistency, reliability, and precision) were developed to assess the efficacy of sweep netting and pan trapping, based on analyses of variations in arthropod abundances, species richness, evenness, capture frequency, and community composition. Neither sampling method was sufficient in any criteria to be used alone for community-level arthropod surveys. On a taxa-specific basis, however, sweep netting was consistent, reliable, and precise for Thysanoptera, infrequently collected (i.e., rare) insects, and Arachnida, whereas pan trapping was consistent, reliable, and precise for Collembola and bees, which is especially significant given current threats to the latter's populations worldwide. Species-level identifications increase the detected dissimilarity between sweep netting and pan trapping. We recommend that community-level arthropod surveys use both sampling methods concurrently, at least in grasslands, but likely in most nonagricultural systems. Target surveys, such as monitoring bee communities in fragmented grassland habitat or where detailed information on behavior of the target arthropod groups is available can in some instances employ singular methods. As a

  1. KM3NeT

    CERN Multimedia

    KM3NeT is a large scale next-generation neutrino telescope located in the deep waters of the Mediterranean Sea, optimized for the discovery of galactic neutrino sources emitting in the TeV energy region.

  2. Understanding Net Zero Energy Buildings

    DEFF Research Database (Denmark)

    Salom, Jaume; Widén, Joakim; Candanedo, José

    2011-01-01

    Although several alternative definitions exist, a Net-Zero Energy Building (Net ZEB) can be succinctly described as a grid-connected building that generates as much energy as it uses over a year. The “net-zero” balance is attained by applying energy conservation and efficiency measures...... and by incorporating renewable energy systems. While based on annual balances, a complete description of a Net ZEB requires examining the system at smaller time-scales. This assessment should address: (a) the relationship between power generation and building loads and (b) the resulting interaction with the power grid....... This paper presents and categorizes quantitative indicators suitable to describe both aspects of the building’s performance. These indicators, named LMGI - Load Matching and Grid Interaction indicators, are easily quantifiable and could complement the output variables of existing building simulation tools...

  3. PolicyNet Publication System

    Data.gov (United States)

    Social Security Administration — The PolicyNet Publication System project will merge the Oracle-based Policy Repository (POMS) and the SQL-Server CAMP system (MSOM) into a new system with an Oracle...

  4. Understanding Net Zero Energy Buildings

    DEFF Research Database (Denmark)

    Salom, Jaume; Widén, Joakim; Candanedo, José

    2011-01-01

    Although several alternative definitions exist, a Net-Zero Energy Building (Net ZEB) can be succinctly described as a grid-connected building that generates as much energy as it uses over a year. The “net-zero” balance is attained by applying energy conservation and efficiency measures...... and by incorporating renewable energy systems. While based on annual balances, a complete description of a Net ZEB requires examining the system at smaller time-scales. This assessment should address: (a) the relationship between power generation and building loads and (b) the resulting interaction with the power grid....... This paper presents and categorizes quantitative indicators suitable to describe both aspects of the building’s performance. These indicators, named LMGI - Load Matching and Grid Interaction indicators, are easily quantifiable and could complement the output variables of existing building simulation tools...

  5. Comparison study of trapped water in human erythrocytes by EPC and DSC method and its application

    Institute of Scientific and Technical Information of China (English)

    ZHAO; Gang; HE; Liqun; GUO; Xiaojie; LIU; Zhong; LUO; Dawei

    2004-01-01

    The novel differential scanning calorimetry method for determining trapped water volume of human red blood cell during freezing process has been reexamined. Results show that the final erythrocyte volume is 53% of its isotonic volume after freezing to -40℃. An electronic particle counter (MultisizerTM III, Beckman Coulter Inc., USA) was used to measure cell volume changes in response to hypertonic solution. Using this approach, when extracellular solution was 3186 mOsm, the equilibrium cell volume was found to be 57% of its isotonic value. Both results indicate that 34%-40% of intracellular water is trapped and cannot respond to osmotic difference between intra- and extracellular solution. These findings are consistent with the published data: at least 20%-32% of the isotonic cell water volume is retained within RBCs during freezing. Some applications of the values of trapped water are addressed.

  6. Proof nets for display logic

    CERN Document Server

    Moot, Richard

    2007-01-01

    This paper explores several extensions of proof nets for the Lambek calculus in order to handle the different connectives of display logic in a natural way. The new proof net calculus handles some recent additions to the Lambek vocabulary such as Galois connections and Grishin interactions. It concludes with an exploration of the generative capacity of the Lambek-Grishin calculus, presenting an embedding of lexicalized tree adjoining grammars into the Lambek-Grishin calculus.

  7. The Net Reclassification Index (NRI)

    DEFF Research Database (Denmark)

    Pepe, Margaret S.; Fan, Jing; Feng, Ziding;

    2015-01-01

    The Net Reclassification Index (NRI) is a very popular measure for evaluating the improvement in prediction performance gained by adding a marker to a set of baseline predictors. However, the statistical properties of this novel measure have not been explored in depth. We demonstrate the alarming...... performance improvement, such as measures derived from the receiver operating characteristic curve, the net benefit function, and the Brier score, cannot be large due to poorly fitting risk functions....

  8. Petri Net Tool Overview 1986

    DEFF Research Database (Denmark)

    Jensen, Kurt; Feldbrugge, Frits

    1987-01-01

    This paper provides an overview of the characteristics of all currently available net based tools. It is a compilation of information provided by tool authors or contact persons. A concise one page overview is provided as well.......This paper provides an overview of the characteristics of all currently available net based tools. It is a compilation of information provided by tool authors or contact persons. A concise one page overview is provided as well....

  9. Process of Petri Nets Extension

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    To describe the dynamic semantics for the network computing, the concept on process is presented based on the semantic model with variable, resource and relation. Accordingly, the formal definition of process and the mapping rules from the specification of Petri nets extension to process are discussed in detail respectively. Based on the collective concepts of process, the specification of dynamic semantics also is constructed as a net system. Finally, to illustrate process intuitively, an example is specified completely.

  10. Oxidation of intramyocellular lipids is dependent on mitochondrial function and the availability of extracellular fatty acids

    DEFF Research Database (Denmark)

    Corpeleijn, Eva; Hessvik, Nina P; Bakke, Siril S;

    2010-01-01

    (2) trapping system and measured under various conditions of extracellular OA (5 or 100 microM) and glucose (0.1 or 5.0 mM) and the absence or presence of mitochondrial uncoupling [carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP)]. First, increased extracellular OA availability (5 vs. 100...... microM) reduced ICL(OX) by 37%. No differences in total lipolysis were observed between low and high OA availability. Uncoupling with FCCP restored ICL(OX) to basal levels during high OA availability. Mitochondrial mass was positively related to ICL(OX), but only in myotubes from lean individuals...

  11. Electron traps in semiconducting polymers: exponential versus Gaussian trap distribution

    NARCIS (Netherlands)

    Nicolai, H.T.; Mandoc, M.M.; Blom, P.W.M.

    2011-01-01

    The low electron currents in poly(dialkoxy-p-phenylene vinylene) (PPV) derivatives and their steep voltage dependence are generally explained by trap-limited conduction in the presence of an exponential trap distribution. Here we demonstrate that the electron transport of several PPV derivatives can

  12. Electron traps in semiconducting polymers : Exponential versus Gaussian trap distribution

    NARCIS (Netherlands)

    Nicolai, H. T.; Mandoc, M. M.; Blom, P. W. M.

    2011-01-01

    The low electron currents in poly(dialkoxy-p-phenylene vinylene) (PPV) derivatives and their steep voltage dependence are generally explained by trap-limited conduction in the presence of an exponential trap distribution. Here we demonstrate that the electron transport of several PPV derivatives can

  13. 29 CFR 4204.13 - Net income and net tangible assets tests.

    Science.gov (United States)

    2010-07-01

    ... 29 Labor 9 2010-07-01 2010-07-01 false Net income and net tangible assets tests. 4204.13 Section....13 Net income and net tangible assets tests. (a) General. The criteria under this section are that either— (1) Net income test. The purchaser's average net income after taxes for its three most...

  14. The Honey Trap

    DEFF Research Database (Denmark)

    Wagner, Michael

    Michael F. Wagner: The Honey Trap –The democratization of leisure through automobilism The automobile has achieved a central position in modern everyday life as an essential artefact to mobility. This raises the question how automobiles have been mediated for mass consumption? The central thesis...... in the article is that the culture of Danish automobilism was constructed around and appropriated through leisure activities conducted primarily by the automobile consumer’s organisation Touring Club de Danemark (FDM). The general purpose for the consumer organisation has been to create a cultural identity...... and a material reality of democratic participation linking ‘Car and Leisure’, a term that has been a central motto for the organization during many decades. The keyword in this activity was ‘Free’ celebrating the manner in which the privately owned automobile secured a maximum of freedom to the owner. The paper...

  15. Efficacy of the Olyset Duo net against insecticide-resistant mosquito vectors of malaria.

    Science.gov (United States)

    Ngufor, Corine; N'Guessan, Raphael; Fagbohoun, Josias; Todjinou, Damien; Odjo, Abibath; Malone, David; Ismail, Hanafy; Akogbeto, Martin; Rowland, Mark

    2016-09-14

    Olyset Duo is a new long-lasting insecticidal net treated with permethrin (a pyrethroid) and pyriproxyfen, an insect growth regulator that disrupts the maturation of oocytes in mosquitoes exposed to the net. We tested the Olyset Duo net against pyrethroid-resistant Anopheles gambiae mosquitoes, which transmit malaria parasites, in laboratory bioassays and in a trial in Benin using experimental huts that closely resemble local habitations. Host-seeking mosquitoes that entered to feed were free to contact the occupied nets and were collected the next morning from exit traps. Surviving blood-fed mosquitoes were observed for effects on reproduction. Control nets were treated with pyrethroid only or pyriproxyfen only, and nets were tested unwashed and after 20 standardized washes. The Olyset Duo net showed improved efficacy and wash resistance relative to the pyrethroid-treated net in terms of mosquito mortality and prevention of blood feeding. The production of offspring among surviving blood-fed A. gambiae in the hut trial was reduced by the pyriproxyfen-treated net and the Olyset Duo net both before washing (90 and 71% reduction, respectively) and after washing (38 and 43% reduction, respectively). The degree of reproductive suppression in the hut trial was predicted by laboratory tunnel tests but not by cone bioassays. The overall reduction in reproductive rate of A. gambiae with the Olyset Duo net in the trial was 94% with no washing and 78% after 20 washes. The Olyset Duo net has the potential to provide community control of mosquito populations and reduce malaria transmission in areas of high insecticide resistance.

  16. Extracellular vesicles: Exosomes, microvesicles, and friends

    NARCIS (Netherlands)

    Raposo, G.; Stoorvogel, W.|info:eu-repo/dai/nl/074352385

    2013-01-01

    Cells release into the extracellular environment diverse types of membrane vesicles of endosomal and plasma membrane origin called exosomes and microvesicles, respectively. These extracellular vesicles (EVs) represent an important mode of intercellular communication by serving as vehicles for

  17. Extracellular enzyme kinetics scale with resource availability

    Science.gov (United States)

    Microbial community metabolism relies on external digestion, mediated by extracellular enzymes that break down complex organic matter into molecules small enough for cells to assimilate. We analyzed the kinetics of 40 extracellular enzymes that mediate the degradation and assimi...

  18. Quantum computing with trapped ions

    Energy Technology Data Exchange (ETDEWEB)

    Hughes, R.J.

    1998-01-01

    The significance of quantum computation for cryptography is discussed. Following a brief survey of the requirements for quantum computational hardware, an overview of the ion trap quantum computation project at Los Alamos is presented. The physical limitations to quantum computation with trapped ions are analyzed and an assessment of the computational potential of the technology is made.

  19. The ALPHA antihydrogen trapping apparatus

    Science.gov (United States)

    Amole, C.; Andresen, G. B.; Ashkezari, M. D.; Baquero-Ruiz, M.; Bertsche, W.; Bowe, P. D.; Butler, E.; Capra, A.; Carpenter, P. T.; Cesar, C. L.; Chapman, S.; Charlton, M.; Deller, A.; Eriksson, S.; Escallier, J.; Fajans, J.; Friesen, T.; Fujiwara, M. C.; Gill, D. R.; Gutierrez, A.; Hangst, J. S.; Hardy, W. N.; Hayano, R. S.; Hayden, M. E.; Humphries, A. J.; Hurt, J. L.; Hydomako, R.; Isaac, C. A.; Jenkins, M. J.; Jonsell, S.; Jørgensen, L. V.; Kerrigan, S. J.; Kurchaninov, L.; Madsen, N.; Marone, A.; McKenna, J. T. K.; Menary, S.; Nolan, P.; Olchanski, K.; Olin, A.; Parker, B.; Povilus, A.; Pusa, P.; Robicheaux, F.; Sarid, E.; Seddon, D.; Seif El Nasr, S.; Silveira, D. M.; So, C.; Storey, J. W.; Thompson, R. I.; Thornhill, J.; Wells, D.; van der Werf, D. P.; Wurtele, J. S.; Yamazaki, Y.

    2014-01-01

    The ALPHA collaboration, based at CERN, has recently succeeded in confining cold antihydrogen atoms in a magnetic minimum neutral atom trap and has performed the first study of a resonant transition of the anti-atoms. The ALPHA apparatus will be described herein, with emphasis on the structural aspects, diagnostic methods and techniques that have enabled antihydrogen trapping and experimentation to be achieved.

  20. Cryogenic silicon surface ion trap

    CERN Document Server

    Niedermayr, Michael; Kumph, Muir; Partel, Stefan; Edlinger, Johannes; Brownnutt, Michael; Blatt, Rainer

    2014-01-01

    Trapped ions are pre-eminent candidates for building quantum information processors and quantum simulators. They have been used to demonstrate quantum gates and algorithms, quantum error correction, and basic quantum simulations. However, to realise the full potential of such systems and make scalable trapped-ion quantum computing a reality, there exist a number of practical problems which must be solved. These include tackling the observed high ion-heating rates and creating scalable trap structures which can be simply and reliably produced. Here, we report on cryogenically operated silicon ion traps which can be rapidly and easily fabricated using standard semiconductor technologies. Single $^{40}$Ca$^+$ ions have been trapped and used to characterize the trap operation. Long ion lifetimes were observed with the traps exhibiting heating rates as low as $\\dot{\\bar{n}}=$ 0.33 phonons/s at an ion-electrode distance of 230 $\\mu$m. These results open many new avenues to arrays of micro-fabricated ion traps.

  1. Accretion discs trapped near corotation

    NARCIS (Netherlands)

    D'Angelo, C.R.; Spruit, H.C.

    2012-01-01

    We show that discs accreting on to the magnetosphere of a rotating star can end up in a trapped state, in which the inner edge of the disc stays near the corotation radius, even at low and varying accretion rates. The accretion in these trapped states can be steady or cyclic; we explore these states

  2. The ALPHA antihydrogen trapping apparatus

    Energy Technology Data Exchange (ETDEWEB)

    Amole, C. [Department of Physics and Astronomy, York University, Toronto ON Canada, M3J 1P3 (Canada); Andresen, G.B. [Department of Physics and Astronomy, Aarhus University, DK-8000 Aarhus C (Denmark); Ashkezari, M.D. [Department of Physics, Simon Fraser University, Burnaby, BC Canada, V5A 1S6 (Canada); Baquero-Ruiz, M. [Department of Physics, University of California at Berkeley, Berkeley, CA 94720-7300 (United States); Bertsche, W. [Department of Physics, College of Science, Swansea University, Swansea SA2 8PP (United Kingdom); School of Physics and Astronomy, University of Manchester, Manchester M13 9PL (United Kingdom); The Cockcroft Institute, Warrington WA4 4AD (United Kingdom); Bowe, P.D. [Department of Physics and Astronomy, Aarhus University, DK-8000 Aarhus C (Denmark); Butler, E. [Physics Department, CERN, CH-1211 Geneva 23 (Switzerland); Capra, A. [Department of Physics and Astronomy, York University, Toronto ON Canada, M3J 1P3 (Canada); Carpenter, P.T. [Department of Physics, Auburn University, Auburn, AL 36849-5311 (United States); Cesar, C.L. [Instituto de Física, Universidade Federal do Rio de Janeiro, Rio de Janeiro 21941-972 (Brazil); Chapman, S. [Department of Physics, University of California at Berkeley, Berkeley, CA 94720-7300 (United States); Charlton, M.; Deller, A.; Eriksson, S. [Department of Physics, College of Science, Swansea University, Swansea SA2 8PP (United Kingdom); Escallier, J. [Brookhaven National Laboratory, Upton, NY 11973 (United States); Fajans, J. [Department of Physics, University of California at Berkeley, Berkeley, CA 94720-7300 (United States); Friesen, T. [Department of Physics and Astronomy, University of Calgary, Calgary AB, Canada, T2N 1N4 (Canada); Fujiwara, M.C.; Gill, D.R. [TRIUMF, 4004 Wesbrook Mall, Vancouver BC, Canada V6T 2A3 (Canada); Gutierrez, A. [Department of Physics and Astronomy, University of British Columbia, Vancouver BC, Canada V6T 1Z4 (Canada); and others

    2014-01-21

    The ALPHA collaboration, based at CERN, has recently succeeded in confining cold antihydrogen atoms in a magnetic minimum neutral atom trap and has performed the first study of a resonant transition of the anti-atoms. The ALPHA apparatus will be described herein, with emphasis on the structural aspects, diagnostic methods and techniques that have enabled antihydrogen trapping and experimentation to be achieved.

  3. Extracellular secretion of recombinant proteins

    Science.gov (United States)

    Linger, Jeffrey G.; Darzins, Aldis

    2014-07-22

    Nucleic acids encoding secretion signals, expression vectors containing the nucleic acids, and host cells containing the expression vectors are disclosed. Also disclosed are polypeptides that contain the secretion signals and methods of producing polypeptides, including methods of directing the extracellular secretion of the polypeptides. Exemplary embodiments include cellulase proteins fused to secretion signals, methods to produce and isolate these polypeptides, and methods to degrade lignocellulosic biomass.

  4. High-level Petri Nets

    DEFF Research Database (Denmark)

    of some of the most important papers on the application and theory of high-level Petri nets. In this way it makes the relevant literature more available. It is our hope that the book will be a useful source of information and that, e.g., it can be used in the organization of Petri net courses. To make...... there is only one kind of token and this means that the state of a place is described by an integer (and in many cases even by a boolean). In high-level nets each token can carry a complex information/data - which, e.g., may describe the entire state of a process or a data base. Today most practical...... by other papers. Thus, e.g., none of the original papers introducing the first versions of high-level Petri nets have been included. The introductions to the individual sections mention a number of researchers who have contributed to the development of high-level Petri nets. Detailed references...

  5. The trapped human experiment.

    Science.gov (United States)

    Huo, R; Agapiou, A; Bocos-Bintintan, V; Brown, L J; Burns, C; Creaser, C S; Devenport, N A; Gao-Lau, B; Guallar-Hoyas, C; Hildebrand, L; Malkar, A; Martin, H J; Moll, V H; Patel, P; Ratiu, A; Reynolds, J C; Sielemann, S; Slodzynski, R; Statheropoulos, M; Turner, M A; Vautz, W; Wright, V E; Thomas, C L P

    2011-12-01

    This experiment observed the evolution of metabolite plumes from a human trapped in a simulation of a collapsed building. Ten participants took it in turns over five days to lie in a simulation of a collapsed building and eight of them completed the 6 h protocol while their breath, sweat and skin metabolites were passed through a simulation of a collapsed glass-clad reinforced-concrete building. Safety, welfare and environmental parameters were monitored continuously, and active adsorbent sampling for thermal desorption GC-MS, on-line and embedded CO, CO(2) and O(2) monitoring, aspirating ion mobility spectrometry with integrated semiconductor gas sensors, direct injection GC-ion mobility spectrometry, active sampling thermal desorption GC-differential mobility spectrometry and a prototype remote early detection system for survivor location were used to monitor the evolution of the metabolite plumes that were generated. Oxygen levels within the void simulator were allowed to fall no lower than 19.1% (v). Concurrent levels of carbon dioxide built up to an average level of 1.6% (v) in the breathing zone of the participants. Temperature, humidity, carbon dioxide levels and the physiological measurements were consistent with a reproducible methodology that enabled the metabolite plumes to be sampled and characterized from the different parts of the experiment. Welfare and safety data were satisfactory with pulse rates, blood pressures and oxygenation, all within levels consistent with healthy adults. Up to 12 in-test welfare assessments per participant and a six-week follow-up Stanford Acute Stress Response Questionnaire indicated that the researchers and participants did not experience any adverse effects from their involvement in the study. Preliminary observations confirmed that CO(2), NH(3) and acetone were effective markers for trapped humans, although interactions with water absorbed in building debris needed further study. An unexpected observation from the NH(3

  6. 1986-87 Annual Trapping Plan

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — This Annual Trapping Plan for the 1986-87 trapping season at Clarence Cannon NWR outlines rules and regulations for the trapping of beaver, muskrat, raccoon,...

  7. MATT: Multi Agents Testing Tool Based Nets within Nets

    Directory of Open Access Journals (Sweden)

    Sara Kerraoui

    2016-12-01

    As part of this effort, we propose a model based testing approach for multi agent systems based on such a model called Reference net, where a tool, which aims to providing a uniform and automated approach is developed. The feasibility and the advantage of the proposed approach are shown through a short case study.

  8. Sublogarithmic uniform Boolean proof nets

    CERN Document Server

    Aubert, Clément

    2012-01-01

    Using a proofs-as-programs correspondence, Terui was able to compare two models of parallel computation: Boolean circuits and proof nets for multiplicative linear logic. Mogbil et. al. gave a logspace translation allowing us to compare their computational power as uniform complexity classes. This paper presents a novel translation in AC0 and focuses on a simpler restricted notion of uniform Boolean proof nets. We can then encode constant-depth circuits and compare complexity classes below logspace, which were out of reach with the previous translations.

  9. Implementing NetScaler VPX

    CERN Document Server

    Sandbu, Marius

    2014-01-01

    An easy-to-follow guide with detailed step-by step-instructions on how to implement the different key components in NetScaler, with real-world examples and sample scenarios.If you are a Citrix or network administrator who needs to implement NetScaler in your virtual environment to gain an insight on its functionality, this book is ideal for you. A basic understanding of networking and familiarity with some of the different Citrix products such as XenApp or XenDesktop is a prerequisite.

  10. Net4Care PHMR Library

    DEFF Research Database (Denmark)

    2014-01-01

    The Net4Care PHMR library contains a) A GreenCDA approach for constructing a data object representing a PHMR document: SimpleClinicalDocument, and b) A Builder which can produce a XML document representing a valid Danish PHMR (following the MedCom profile) document from the SimpleClinicalDocument......The Net4Care PHMR library contains a) A GreenCDA approach for constructing a data object representing a PHMR document: SimpleClinicalDocument, and b) A Builder which can produce a XML document representing a valid Danish PHMR (following the MedCom profile) document from the Simple...

  11. Development of a gravid trap for collecting live malaria vectors Anopheles gambiae s.l.

    Directory of Open Access Journals (Sweden)

    Sisay Dugassa

    Full Text Available BACKGROUND: Effective malaria vector control targeting indoor host-seeking mosquitoes has resulted in fewer vectors entering houses in many areas of sub-Saharan Africa, with the proportion of vectors outdoors becoming more important in the transmission of this disease. This study aimed to develop a gravid trap for the outdoor collection of the malaria vector Anopheles gambiae s.l. based on evaluation and modification of commercially available gravid traps. METHODS: Experiments were implemented in an 80 m(2 semi-field system where 200 gravid Anopheles gambiae s.s. were released nightly. The efficacy of the Box, CDC and Frommer updraft gravid traps was compared. The Box gravid trap was tested to determine if the presence of the trap over water and the trap's sound affected catch size. Mosquitoes approaching the treatment were evaluated using electrocuting nets or detergents added to the water in the trap. Based on the results, a new gravid trap (OviART trap that provided an open, unobstructed oviposition site was developed and evaluated. RESULTS: Box and CDC gravid traps collected similar numbers (relative rate (RR 0.8, 95% confidence interval (CI 0.6-1.2; p = 0.284, whereas the Frommer trap caught 70% fewer mosquitoes (RR 0.3, 95% CI 0.2-0.5; p < 0.001. The number of mosquitoes approaching the Box trap was significantly reduced when the trap was positioned over a water-filled basin compared to an open pond (RR 0.7 95% CI 0.6-0.7; p < 0.001. This effect was not due to the sound of the trap. Catch size increased by 60% (RR 1.6, 1.2-2.2; p = 0.001 with the new OviART trap. CONCLUSION: Gravid An. Gambiae s.s. females were visually deterred by the presence of the trapping device directly over the oviposition medium. Based on these investigations, an effective gravid trap was developed that provides open landing space for egg-laying Anopheles.

  12. First Attempts at Antihydrogen Trapping in ALPHA

    Science.gov (United States)

    Andresen, G. B.; Bertsche, W.; Bowe, P. D.; Bray, C. C.; Butler, E.; Cesar, C. L.; Chapman, S.; Charlton, M.; Fajans, J.; Fujiwara, M. C.; Funakoshi, R.; Gill, D. R.; Hangst, J. S.; Hardy, W. N.; Hayano, R. S.; Hayden, M. E.; Humphries, A. J.; Hydomako, R.; Jenkins, M. J.; Jørgensen, L. V.; Kurchaninov, L.; Lambo, R.; Madsen, N.; Nolan, P.; Olchanski, K.; Olin, A.; Page, R. D.; Povilus, A.; Pusa, P.; Robicheaux, F.; Sarid, E.; El Nasr, S. Seif; Silveira, D. M.; Storey, J. W.; Thompson, R. I.; van der Werf, D. P.; Wurtele, J. S.; Yamazaki, Y.

    2008-08-01

    The ALPHA apparatus is designed to produce and trap antihydrogen atoms. The device comprises a multifunction Penning trap and a superconducting, neutral atom trap having a minimum-B configuration. The atom trap features an octupole magnet for transverse confinement and solenoidal mirror coils for longitudinal confinement. The magnetic trap employs a fast shutdown system to maximize the probability of detecting the annihilation of released antihydrogen. In this article we describe the first attempts to observe antihydrogen trapping.

  13. Feedback traps for virtual potentials

    CERN Document Server

    Gavrilov, Momčilo

    2016-01-01

    Feedback traps are tools for trapping and manipulating single charged objects, such as molecules in solution. An alternative to optical tweezers and other single-molecule techniques, they use feedback to counteract the Brownian motion of a molecule of interest. The trap first acquires information about a molecule's position and then applies an electric feedback force to move the molecule. Since electric forces are stronger than optical forces at small scales, feedback traps are the best way to trap single molecules without "touching" them. Feedback traps can do more than trap molecules: They can also subject a target object to forces that are calculated to be the gradient of a desired potential function U(x). If the feedback loop is fast enough, it creates a virtual potential whose dynamics will be very close to those of a particle in an actual potential U(x). But because the dynamics are entirely a result of the feedback loop--absent the feedback, there is only an object diffusing in a fluid--we are free to ...

  14. Trapping tsetse flies on water

    Directory of Open Access Journals (Sweden)

    Laveissière C.

    2011-05-01

    Full Text Available Riverine tsetse flies such as Glossina palpalis gambiensis and G. tachinoides are the vectors of human and animal trypanosomoses in West Africa. Despite intimate links between tsetse and water, to our knowledge there has never been any attempt to design trapping devices that would catch tsetse on water. In mangrove (Guinea one challenging issue is the tide, because height above the ground for a trap is a key factor affecting tsetse catches. The trap was mounted on the remains of an old wooden dugout, and attached with rope to nearby branches, thereby allowing it to rise and fall with the tide. Catches showed a very high density of 93.9 flies/”water-trap”/day, which was significantly higher (p < 0.05 than all the catches from other habitats where the classical trap had been used. In savannah, on the Comoe river of South Burkina Faso, the biconical trap was mounted on a small wooden raft anchored to a stone, and catches were compared with the classical biconical trap put on the shores. G. p. gambiensis and G. tachinoides densities were not significantly different from those from the classical biconical one. The adaptations described here have allowed to efficiently catch tsetse on the water, which to our knowledge is reported here for the first time. This represents a great progress and opens new opportunities to undertake studies on the vectors of trypanosomoses in mangrove areas of Guinea, which are currently the areas showing the highest prevalences of sleeping sickness in West Africa. It also has huge potential for tsetse control using insecticide impregnated traps in savannah areas where traps become less efficient in rainy season. The Guinean National control programme has already expressed its willingness to use such modified traps in its control campaigns in Guinea, as has the national PATTEC programme in Burkina Faso during rainy season.

  15. A reservoir trap for antiprotons

    CERN Document Server

    Smorra, Christian; Franke, Kurt; Nagahama, Hiroki; Schneider, Georg; Higuchi, Takashi; Van Gorp, Simon; Blaum, Klaus; Matsuda, Yasuyuki; Quint, Wolfgang; Walz, Jochen; Yamazaki, Yasunori; Ulmer, Stefan

    2015-01-01

    We have developed techniques to extract arbitrary fractions of antiprotons from an accumulated reservoir, and to inject them into a Penning-trap system for high-precision measurements. In our trap-system antiproton storage times > 1.08 years are estimated. The device is fail-safe against power-cuts of up to 10 hours. This makes our planned comparisons of the fundamental properties of protons and antiprotons independent from accelerator cycles, and will enable us to perform experiments during long accelerator shutdown periods when background magnetic noise is low. The demonstrated scheme has the potential to be applied in many other precision Penning trap experiments dealing with exotic particles.

  16. Pattern formation with trapped ions

    CERN Document Server

    Lee, Tony E

    2010-01-01

    We propose an experiment to study collective behavior in a nonlinear medium of trapped ions. Using laser cooling and heating and an anharmonic trap potential, one can turn an ion into a nonlinear van der Pol-Duffing oscillator. A chain of ions interacting electrostatically has stable plane waves for all parameters. The system also behaves like an excitable medium, since a sufficiently large perturbation generates a travelling pulse. Small chains exhibit multistability and limit cycles. We account for noise from spontaneous emission in the amplitude equation and find that the patterns are observable for realistic experimental parameters. The tunability of ion traps makes them an exciting setting to study nonequilibrium statistical physics.

  17. Candidate gene analysis of the human natural killer-1 carbohydrate pathway and perineuronal nets in schizophrenia: B3GAT2 is associated with disease risk and cortical surface area

    DEFF Research Database (Denmark)

    Kähler, Anna K; Djurovic, Srdjan; Rimol, Lars M

    2011-01-01

    The Human Natural Killer-1 carbohydrate (HNK-1) is involved in neurodevelopment and synaptic plasticity. Extracellular matrix structures called perineuronal nets, condensed around subsets of neurons and proximal dendrites during brain maturation, regulate synaptic transmission and plasticity....

  18. A New Atom Trap The Annular Shell Atom Trap (ASAT)

    CERN Document Server

    Pilloff, H S; Pilloff, Herschel S.; Horbatsch, Marko

    2002-01-01

    In the course of exploring some aspects of atom guiding in a hollow, optical fiber, a small negative potential energy well was found just in front of the repulsive or guiding barrier. This results from the optical dipole and the van der Waals potentials. The ground state for atoms bound in this negative potential well was determined by numerically solving the Schrodinger eq. and it was found that this negative well could serve as an atom trap. This trap is referred to as the Annular Shell Atom Trap or ASAT because of the geometry of the trapped atoms which are located in the locus of points defining a very thin annular shell just in front of the guiding barrier. A unique feature of the ASAT is the compression of the atoms from the entire volume to the volume of the annular shell resulting in a very high density of atoms in this trap. This trap may have applications to very low temperatures using evaporative cooling and possibly the formation of BEC. Finally, a scheme is discussed for taking advantage of the d...

  19. Surgery for GEP-NETs

    DEFF Research Database (Denmark)

    Knigge, Ulrich; Hansen, Carsten Palnæs

    2012-01-01

    Surgery is the only treatment that may cure the patient with gastroentero-pancreatic (GEP) neuroendocrine tumours (NET) and neuroendocrine carcinomas (NEC) and should always be considered as first line treatment if R0/R1 resection can be achieved. The surgical and interventional procedures for GEP...

  20. The Net Generation Cheating Challenge

    Science.gov (United States)

    Milliron, Valerie; Sandoe, Kent

    2008-01-01

    Integral to higher education, academic integrity stands as a cornerstone of academic life. However, compelling evidence of widespread academic dishonesty among Net-Generation students threatens to undermine both the environment of trust that nourishes integrity and the safeguards that help ensure it. Working from their experience with widespread…

  1. Reference Guide Microsoft.NET

    NARCIS (Netherlands)

    Zee M van der; Verspaij GJ; Rosbergen S; IMP; NMD

    2003-01-01

    Met behulp van het rapport kunnen ontwikkelaars, beheerders en betrokken managers bij ICT projecten meer inzicht krijgen in de .NET technologie en een goede keuze maken in de inzetbaarheid van deze technologie. Het rapport geeft de bevindingen en conclusies van een verkennende studie naar het nu

  2. Reference Guide Microsoft.NET

    NARCIS (Netherlands)

    Zee M van der; Verspaij GJ; Rosbergen S; IMP; NMD

    2003-01-01

    Met behulp van het rapport kunnen ontwikkelaars, beheerders en betrokken managers bij ICT projecten meer inzicht krijgen in de .NET technologie en een goede keuze maken in de inzetbaarheid van deze technologie. Het rapport geeft de bevindingen en conclusies van een verkennende studie naar het

  3. Reference Guide Microsoft.NET

    NARCIS (Netherlands)

    Zee M van der; Verspaij GJ; Rosbergen S; IMP; NMD

    2003-01-01

    Met behulp van het rapport kunnen ontwikkelaars, beheerders en betrokken managers bij ICT projecten meer inzicht krijgen in de .NET technologie en een goede keuze maken in de inzetbaarheid van deze technologie. Het rapport geeft de bevindingen en conclusies van een verkennende studie naar het nu

  4. Petri Nets in Cryptographic Protocols

    DEFF Research Database (Denmark)

    Crazzolara, Federico; Winskel, Glynn

    2001-01-01

    A process language for security protocols is presented together with a semantics in terms of sets of events. The denotation of process is a set of events, and as each event specifies a set of pre and postconditions, this denotation can be viewed as a Petri net. By means of an example we illustrat...

  5. Communicating with the Net Generation

    Science.gov (United States)

    2011-03-11

    narcissistic ―me‖ generation to a perceived lack of respect for authority. The Net Generation displays high self-esteem and confidence but appear much more...by our adversaries, especially by terrorist organizations that continually look for vulnerabilities to exploit. Location- based social networking

  6. Inhibition of PAD4 activity is sufficient to disrupt mouse and human NET formation

    Science.gov (United States)

    Lewis, Huw D.; Liddle, John; Coote, Jim E.; Atkinson, Stephen J.; Barker, Michael D.; Bax, Benjamin, D.; Bicker, Kevin L.; Bingham, Ryan P.; Campbell, Matthew; Chen, Yu Hua; Chung, Chun-wa; Craggs, Peter D.; Davis, Rob P.; Eberhard, Dirk; Joberty, Gerard; Lind, Kenneth E.; Locke, Kelly; Maller, Claire; Martinod, Kimberly; Patten, Chris; Polyakova, Oxana; Rise, Cecil E.; Rüdiger, Martin; Sheppard, Robert J.; Slade, Daniel J.; Thomas, Pamela; Thorpe, Jim; Yao, Gang; Drewes, Gerard; Wagner, Denisa D.; Thompson, Paul R.; Prinjha, Rab K.; Wilson, David M.

    2015-01-01

    PAD4 has been strongly implicated in the pathogenesis of autoimmune, cardiovascular and oncological diseases, through clinical genetics and gene disruption in mice. Novel, selective PAD4 inhibitors binding to a calcium-deficient form of the PAD4 enzyme have, for the first time, validated the critical enzymatic role of human and mouse PAD4 in both histone citrullination and neutrophil extracellular trap formation. The therapeutic potential of PAD4 inhibitors can now be explored. PMID:25622091

  7. Innovation: the classic traps.

    Science.gov (United States)

    Kanter, Rosabeth Moss

    2006-11-01

    these traps.

  8. Trap closure and prey retention in Venus flytrap (Dionaea muscipula) temporarily reduces photosynthesis and stimulates respiration.

    Science.gov (United States)

    Pavlovic, Andrej; Demko, Viktor; Hudák, Ján

    2010-01-01

    The carnivorous plant Venus flytrap (Dionaea muscipula) produces a rosette of leaves: each leaf is divided into a lower part called the lamina and an upper part, the trap, with sensory trigger hairs on the adaxial surface. The trap catches prey by very rapid closure, within a fraction of a second of the trigger hairs being touched twice. Generation of action potentials plays an important role in closure. Because electrical signals are involved in reduction of the photosynthetic rate in different plant species, we hypothesized that trap closure and subsequent movement of prey in the trap will result in transient downregulation of photosynthesis, thus representing the energetic costs of carnivory associated with an active trapping mechanism, which has not been previously described. Traps were enclosed in a gas exchange cuvette and the trigger hairs irritated with thin wire, thus simulating insect capture and retention. Respiration rate was measured in darkness (RD). In the light, net photosynthetic rate (AN), stomatal conductance (gs) and intercellular CO2 concentration (ci) were measured, combined with chlorophyll fluorescence imaging. Responses were monitored in the lamina and trap separately. Irritation of trigger hairs resulted in decreased AN and increased RD, not only immediately after trap closure but also during the subsequent period when prey retention was simulated in the closed trap. Stomatal conductance remained stable, indicating no stomatal limitation of AN, so ci increased. At the same time, the effective quantum yield of photosystem II (PSII) decreased transiently. The response was confined mainly to the digestive zone of the trap and was not observed in the lamina. Stopping mechanical irritation resulted in recovery of AN, RD and PSII. We put forward the first experimental evidence for energetic demands and carbon costs during insect trapping and retention in carnivorous plants, providing a new insight into the cost/benefit model of carnivory.

  9. 26 CFR 1.172-4 - Net operating loss carrybacks and net operating loss carryovers.

    Science.gov (United States)

    2010-04-01

    ... 26 Internal Revenue 3 2010-04-01 2010-04-01 false Net operating loss carrybacks and net operating... Corporations (continued) § 1.172-4 Net operating loss carrybacks and net operating loss carryovers. (a) General provisions—(1) Years to which loss may be carried—(i) In general. In order to compute the net operating...

  10. Trapping Triatominae in Silvatic Habitats

    Directory of Open Access Journals (Sweden)

    Noireau François

    2002-01-01

    Full Text Available Large-scale trials of a trapping system designed to collect silvatic Triatominae are reported. Live-baited adhesive traps were tested in various ecosystems and different triatomine habitats (arboreal and terrestrial. The trials were always successful, with a rate of positive habitats generally over 20% and reaching 48.4% for palm trees of the Amazon basin. Eleven species of Triatominae belonging to the three genera of public health importance (Triatoma, Rhodnius and Panstrongylus were captured. This trapping system provides an effective way to detect the presence of triatomines in terrestrial and arboreal silvatic habitats and represents a promising tool for ecological studies. Various lines of research are contemplated to improve the performance of this trapping system.

  11. Magneto optical trapping of Barium

    CERN Document Server

    De, S; Jungmann, K; Willmann, L

    2008-01-01

    First laser cooling and trapping of the heavy alkaline earth element barium has been achieved based on the strong 6s$^2$ $^1$S$_0$ - 6s6p $^1$P$_1$ transition for the main cooling. Due to the large branching into metastable D-states several additional laser driven transitions are required to provide a closed cooling cycle. A total efficiency of $0.4(1) \\cdot 10^{-2}$ for slowing a thermal atomic beam and capturing atoms into a magneto optical trap was obtained. Trapping lifetimes of more than 1.5 s were observed. This lifetime is shortened at high laser intensities by photo ionization losses. The developed techniques will allow to extend significantly the number of elements that can be optically cooled and trapped.

  12. Seismic fault zone trapped noise

    National Research Council Canada - National Science Library

    Hillers, G; Campillo, M; Ben‐Zion, Y; Roux, P

    2014-01-01

    Systematic velocity contrasts across and within fault zones can lead to head and trapped waves that provide direct information on structural units that are important for many aspects of earthquake and fault mechanics...

  13. Extracellular nucleotide signaling in plants

    Energy Technology Data Exchange (ETDEWEB)

    Stacey, Gary [Univ. of Missouri, Columbia, MO (United States)

    2016-09-08

    Over the life of this funded project, our research group identified and characterized two key receptor proteins in plants; one mediating the innate immunity response to chitin and the other elucidating the key receptor for extracellular ATP. In the case of chitin recognition, we recently described the quaternary structure of this receptor, shedding light on how the receptor functions. Perhaps more importantly, we demonstrated that all plants have the ability to recognize both chitin oligomers and lipochitooligosacchardes, fundamentally changing how the community views the evolution of these systems and strategies that might be used, for example, to extend symbiotic nitrogen fixation to non-legumes. Our discovery of DORN1 opens a new chapter in plant physiology documenting conclusively that eATP is an important extracellular signal in plants, as it is in animals. At this point, we cannot predict just how far reaching this discovery may prove to be but we are convinced that eATP signaling is fundamental to plant growth and development and, hence, we believe that the future will be very exciting for the study of DORN1 and its overall function in plants.

  14. Single cell cloning and recombinant monoclonal antibodies generation from RA synovial B cells reveal frequent targeting of citrullinated histones of NETs

    Science.gov (United States)

    Corsiero, Elisa; Bombardieri, Michele; Carlotti, Emanuela; Pratesi, Federico; Robinson, William; Migliorini, Paola; Pitzalis, Costantino

    2016-01-01

    Objectives Rheumatoid arthritis (RA) is characterised by breach of self-tolerance towards citrullinated antigens with generation of anti-citrullinated peptide/proteins antibodies (ACPA). Currently, the nature and source of citrullinated antigens driving the humoral autoimmune response within synovial ectopic lymphoid structures (ELS) is a crucial unknown aspect of RA pathogenesis. Here we characterised the autoreactive B-cell response of lesional B cells isolated from ELS+RA synovium. Methods Single synovial tissue CD19+cells were Fluorescence Activated Cell Sorting (FACS)-sorted and VH/VL Ig genes cloned to generate recombinant monoclonal antibodies (rmAbs) from patients with ELS+/ACPA+RA. Results RA-rmAbs immunoreactivity analysis provided the following key findings: (1) in a chIP-based array containing 300 autoantigens and in a ‘citrullinome’ multiplex assay, a strong reactivity against citrullinated histones H2A/H2B (citH2A/H2B) was observed in ∼40% of RA-rmAbs, followed by cit-fibrinogen and cit-vimentin; (2) anti-citH2A/H2B-reactive RA-rmAbs (but not anti-citH2A/H2B negative) selectively recognised neutrophil extracellular traps (NETs) from peripheral blood and/or RA joint neutrophils; (3) anti-citH2A/citH2B and anti-NET immunobinding was dependent on affinity maturation and was completely abrogated following reversion of hypermutated IgVH/VL genes to germline sequences; (4) ELS+ (not ELS−) RA synovial tissues engrafted into Severe Combined ImmunoDeficiency (SCID) mice released human anti-citH2A/citH2B and anti-NET antibodies in association with the intra-graft expression of CXCL13 and lymphotoxin (LT)-β, two master regulators of ELS. Conclusion We provided novel evidence that B cells differentiated within synovial ELS in the RA joints frequent target deiminated proteins which could be generated during NETosis of RA synovial neutrophils including histones. Thus, NETs could represent a source of citrullinated antigens fuelling the ACPA autoimmune

  15. Army Net Zero Prove Out. Army Net Zero Training Report

    Science.gov (United States)

    2014-11-20

    reclaim system does not enable Fort Carson to achieve its net zero water goal, which is to reclaim water through the WWTP equal to or greater than the...Class A Recycled Water Plant WWTP rapid infiltration basin #1 (of 4) 426,000 gal water supply storage reservoir Primary Lagoon for WWTP 6 7...cost of water = Longer time to payback  Double charged for incoming and outgoing  Army owned WWTP – true cost of water = water/sewer rates It

  16. Army Net Zero Prove Out. Integrated Net Zero Best Practices

    Science.gov (United States)

    2014-11-18

    Program Office (Office of the Assistant Secretary of the Army for Installations and Environment) 5850 21st Street, Building 211, Second Floor Fort...of energy, water, and waste to capture and commercialize the resource value and/or enhance the ecological productivity of land, water, and air. A...water, and waste to capture and commercialize the resource value and/or enhance the ecological productivity of land, water, and air. A Net Zero

  17. Process expression of bounded Petri nets

    Institute of Scientific and Technical Information of China (English)

    吴哲辉

    1996-01-01

    The concept of process expression of bounded Petri nets is presented.Moreover,an algorithm to find the process expression for a bounded Petri net is given.A process expression of a bounded Petri net is a regular expression whose every alphabet symbol represents a basic subprocess of the net.The regular set expressed by the regular expression is the set of all surjective processes of a bounded Petri net.A surjective process of a bounded Petri net is a process of this net in which every s-cut corresponds to a reachable marking of the net.Therefore,all surjective processes of a bounded Petri net can be obtained as long as its process expression and the basic subprocess represented by the alphabet symbols of the process expression are given.

  18. Are mist nets a menace for threatened insects? A first attempt to gather quantitative data

    DEFF Research Database (Denmark)

    Carpaneto, Giuseppe Maria; Mazziotta, Adriano; Maltzeff, Paolo

    2011-01-01

    level and represent a bycatch of mist net techniques, especially in early spring and at sunrise, when their swarming and reproductive activity is high. When summed with other factors affecting these beetles, such as habitat loss and fragmentation, this tool may represent a threat for vulnerable insect...... populations. Trapping effects may also represent a depletion of food resource for some insectivorous birds. For these reasons we recommend cooperative research between ornithologists and entomologists to fully study the bycatch effects of mist netting on flying insects and their effects on food availability...

  19. Capturing birds with mist nets: A review

    Science.gov (United States)

    Keyes, B.E.; Grue, C.E.

    1982-01-01

    Herein we have tried to provide a comprehensive review of mist-netting techniques suitable for both novice and experienced netters. General mist-netting procedures and modifications developed by netters for particular bird species and habitats are included. Factors which influence capture success, including site selection, net specifications and placement, weather, and time of day, are discussed. Guidelines are presented for the care of netted birds and the use of mist-net data in the study of bird communities. The advantages of the use of mist nets over other methods of capturing birds are also discussed.

  20. Complexity Metrics for Workflow Nets

    DEFF Research Database (Denmark)

    Lassen, Kristian Bisgaard; van der Aalst, Wil M.P.

    2009-01-01

    , etc. It seems obvious that the complexity of the model contributes to design errors and a lack of understanding. It is not easy to measure complexity, however. This paper presents three complexity metrics that have been implemented in the process analysis tool ProM. The metrics are defined...... analysts have difficulties grasping the dynamics implied by a process model. Recent empirical studies show that people make numerous errors when modeling complex business processes, e.g., about 20 percent of the EPCs in the SAP reference model have design flaws resulting in potential deadlocks, livelocks...... for a subclass of Petri nets named Workflow nets, but the results can easily be applied to other languages. To demonstrate the applicability of these metrics, we have applied our approach and tool to 262 relatively complex Protos models made in the context of various student projects. This allows us to validate...

  1. Net4Care PHMR Library

    DEFF Research Database (Denmark)

    2014-01-01

    The Net4Care PHMR library contains a) A GreenCDA approach for constructing a data object representing a PHMR document: SimpleClinicalDocument, and b) A Builder which can produce a XML document representing a valid Danish PHMR (following the MedCom profile) document from the SimpleClinicalDocument......The Net4Care PHMR library contains a) A GreenCDA approach for constructing a data object representing a PHMR document: SimpleClinicalDocument, and b) A Builder which can produce a XML document representing a valid Danish PHMR (following the MedCom profile) document from the Simple......ClinicalDocument object. This software component is adopted by several Danish companies as PHMR engine for telemedical applications, and is used in the process of writing the Danish PHMR standard....

  2. Behavior Relativity of Petri Nets

    Institute of Scientific and Technical Information of China (English)

    蒋昌俊; 王怀清; 廖少毅

    2002-01-01

    This paper presents a novel methodology for modelling and analyzing of behav-ior relations of concurrent systems. The set of all firing sequences of a Petri net is an importanttool for describing the dynamic behavior of concurrent systems. In this paper, the behaviorrelativity of two concurrent subsystems in their synchronous composition is presented. Such be-havior relativities, including Controlled Relativity, United Relativity, Interactive Relativity andExclusive Relativity, are defined respectively. The properties of the relativities are discussed indetail. The analysis method for these properties is based on minimum T-invariants, when twosubsystems are live bounded Petri nets. A well-known example has also been analyaed usingthe new methodology to demonstrate the advantages of the proposed methodology.

  3. Prosocial behavior on the Net.

    Science.gov (United States)

    Sproull, Lee

    2011-01-01

    Volunteers and charitable organizations contribute significantly to community welfare through their prosocial behavior: that is, discretionary behavior such as assisting, comforting, sharing, and cooperating intended to help worthy beneficiaries. This essay focuses on prosocial behavior on the Internet. It describes how offline charitable organizations are using the Net to become more efficient and effective. It also considers entirely new models of Net-based volunteer behavior directed at creating socially beneficial information goods and services. After exploring the scope and diversity of online prosocial behavior, the essay focuses on ways to encourage this kind of behavior through appropriate task and social structures, motivational signals, and trust indicators. It concludes by asking how local offline communities ultimately could be diminished or strengthened as prosocial behavior increases online.

  4. Microinstrument gradient-force optical trap.

    Science.gov (United States)

    Collins, S D; Baskin, R J; Howitt, D G

    1999-10-01

    A micromachined fiber-optic trap is presented. The trap consists of four single-mode, 1064-nm optical intersection. The beam fibers mounted in a micromachined silicon and glass housing. Micromachining provides the necessary precision to align the four optical fibers so that the outputs have a common intersection forms a strong three-dimensional gradient-force trap with trapping forces comparable with that of optical tweezers. Characterization of the multibeam fiber trap is illustrated for capture of polystyrene microspheres, computer simulations of the trap stiffness, and experimental determination of the trapping forces.

  5. Genomic mechanisms accounting for the adaptation to parasitism in nematode-trapping fungi.

    Directory of Open Access Journals (Sweden)

    Tejashwari Meerupati

    2013-11-01

    Full Text Available Orbiliomycetes is one of the earliest diverging branches of the filamentous ascomycetes. The class contains nematode-trapping fungi that form unique infection structures, called traps, to capture and kill free-living nematodes. The traps have evolved differently along several lineages and include adhesive traps (knobs, nets or branches and constricting rings. We show, by genome sequencing of the knob-forming species Monacrosporium haptotylum and comparison with the net-forming species Arthrobotrys oligospora, that two genomic mechanisms are likely to have been important for the adaptation to parasitism in these fungi. Firstly, the expansion of protein domain families and the large number of species-specific genes indicated that gene duplication followed by functional diversification had a major role in the evolution of the nematode-trapping fungi. Gene expression indicated that many of these genes are important for pathogenicity. Secondly, gene expression of orthologs between the two fungi during infection indicated that differential regulation was an important mechanism for the evolution of parasitism in nematode-trapping fungi. Many of the highly expressed and highly upregulated M. haptotylum transcripts during the early stages of nematode infection were species-specific and encoded small secreted proteins (SSPs that were affected by repeat-induced point mutations (RIP. An active RIP mechanism was revealed by lack of repeats, dinucleotide bias in repeats and genes, low proportion of recent gene duplicates, and reduction of recent gene family expansions. The high expression and rapid divergence of SSPs indicate a striking similarity in the infection mechanisms of nematode-trapping fungi and plant and insect pathogens from the crown groups of the filamentous ascomycetes (Pezizomycotina. The patterns of gene family expansions in the nematode-trapping fungi were more similar to plant pathogens than to insect and animal pathogens. The observation

  6. Net sustainable buildings: Approaching future

    Science.gov (United States)

    Bojić, Milorad

    2012-11-01

    For a net sustainable building (NSB), the paper develops its definition, and gives its characteristics. The NSB should provide the maximum thermal comfort were its energy consumption is totally covered by energy of renewable origin during its life cycle. The covered energy consumption should be operational and embodied. The used renewable energy may be that of sun and of anergy. The building may be furthermore evaluated regarding its status with regard to the mismatch, primary energy, emergy, exergy, fossil energy, carbon and dissemination.

  7. 1990 Joint Military Net Assessment

    Science.gov (United States)

    2007-11-02

    the Office of Net Assessment 6 Deputy Director for Theater Assessments and Planning, Office of the Assistant Secretary of Defense for Program Analysis ...countermeasures electronic countermeasures European Economic Community electronic warfare -F- FOFA FY follow-on-forces attack fiscal year -G... analysis of current and projected capabilities of US forces, assisted by allies where appropriate, to deter war and, if deterrence fails, to terminate the

  8. Live trapping of hawks and owls

    Science.gov (United States)

    Stewart, R.E.; Cope, J.B.; Robbins, C.S.

    1945-01-01

    1. Hawks of six species (80 individuals) and owls of five species (37 individuals) were trapped for banding from November 1, 1943, to. May 26,1944. 2. In general, pole traps proved better than hand-operated traps or automatic traps using live bait. 3. Verbail pole traps proved very efficient, and were much more humane than padded steel traps because they rarely injured a captured bird. 4: Unbaited Verbail traps took a variety of raptors, in rough proportion to their local abundance, although slightly more of beneficial species were caught than of harmful types. 5. Hawks and owls were retrapped more readily in Verbail traps than in other types tried. 6. The number of song birds caught in Verbail traps was negligible. 7. Crows and vultures were not taken in Verbail traps, but possibly could be caught with bait.

  9. Advanced Near Net Shape Technology

    Science.gov (United States)

    Vickers, John

    2015-01-01

    The objective of the Advanced Near Net Shape Technology (ANNST) project is to radically improve near net shape manufacturing methods from the current Technology/ Manufacturing Readiness Levels (TRL/MRL 3-4) to the point where they are viable candidates (TRL/ MRL-6) for shortening the time and cost for insertion of new aluminum alloys and revolutionary manufacturing methods into the development/improvement of space structures. Conventional cyrotank manufacturing processes require fabrication of multiple pieces welded together to form a complete tank. A variety of near net shape manufacturing processes has demonstrated excellent potential for enabling single-piece construction of components such as domes, barrels, and ring frames. Utilization of such processes can dramatically reduce the extent of welding and joining needed to construct cryogenic tanks and other aerospace structures. The specific focus of this project is to successfully mature the integrally stiffened cylinder (ISC) process in which a single-piece cylinder with integral stiffeners is formed in one spin/flow forming process. Structural launch vehicle components, like cryogenic fuel tanks (e.g., space shuttle external tank), are currently fabricated via multipiece assembly of parts produced through subtractive manufacturing techniques. Stiffened structural panels are heavily machined from thick plate, which results in excessive scrap rates. Multipiece construction requires welds to assemble the structure, which increases the risk for defects and catastrophic failures.

  10. Activation of perineuronal net-expressing excitatory neurons during associative memory encoding and retrieval

    Science.gov (United States)

    Morikawa, Shota; Ikegaya, Yuji; Narita, Minoru; Tamura, Hideki

    2017-01-01

    Perineuronal nets (PNNs), proteoglycan-rich extracellular matrix structures, are thought to be expressed around inhibitory neurons and contribute to critical periods of brain function and synaptic plasticity. However, in some specific brain regions such as the amygdala, PNNs were predominantly expressed around excitatory neurons. These neurons were recruited during auditory fear conditioning and memory retrieval. Indeed, the activation of PNN-expressing excitatory neurons predicted cognitive performance. PMID:28378772

  11. Analysis of extracellular RNA by digital PCR

    Directory of Open Access Journals (Sweden)

    Kenji eTakahashi

    2014-06-01

    Full Text Available The transfer of extracellular RNA is emerging as an important mechanism for intracellular communication. The ability for the transfer of functionally active RNA molecules from one cell to another within vesicles such as exosomes enables a cell to modulate cellular signaling and biological processes within recipient cells. The study of extracellular RNA requires sensitive methods for the detection of these molecules. In this methods article, we will describe protocols for the detection of such extracellular RNA using sensitive detection technologies such as digital PCR. These protocols should be valuable to researchers interested in the role and contribution of extracellular RNA to tumor cell biology.

  12. Extracellular Molecules Involved in Cancer Cell Invasion

    Energy Technology Data Exchange (ETDEWEB)

    Stivarou, Theodora; Patsavoudi, Evangelia, E-mail: epatsavoudi@pasteur.gr [Department of Biochemistry, Hellenic Pasteur Institute, Athens 11521 (Greece); Technological Educational Institute of Athens, Egaleo, Athens 12210 (Greece)

    2015-01-26

    Nowadays it is perfectly clear that understanding and eradicating cancer cell invasion and metastasis represent the crucial, definitive points in cancer therapeutics. During the last two decades there has been a great interest in the understanding of the extracellular molecular mechanisms involved in cancer cell invasion. In this review, we highlight the findings concerning these processes, focusing in particular on extracellular molecules, including extracellular matrix proteins and their receptors, growth factors and their receptors, matrix metalloproteinases and extracellular chaperones. We report the molecular mechanisms underlying the important contribution of this pool of molecules to the complex, multi-step phenomenon of cancer cell invasion.

  13. Extracellular Molecules Involved in Cancer Cell Invasion

    Directory of Open Access Journals (Sweden)

    Theodora Stivarou

    2015-01-01

    Full Text Available Nowadays it is perfectly clear that understanding and eradicating cancer cell invasion and metastasis represent the crucial, definitive points in cancer therapeutics. During the last two decades there has been a great interest in the understanding of the extracellular molecular mechanisms involved in cancer cell invasion. In this review, we highlight the findings concerning these processes, focusing in particular on extracellular molecules, including extracellular matrix proteins and their receptors, growth factors and their receptors, matrix metalloproteinases and extracellular chaperones. We report the molecular mechanisms underlying the important contribution of this pool of molecules to the complex, multi-step phenomenon of cancer cell invasion.

  14. A magnetic particle micro-trap for large trapping surfaces

    KAUST Repository

    Gooneratne, Chinthaka P.

    2012-01-08

    Manipulation of micron-size magnetic particles of the superparamagnetic type contributes significantly in many applications like controlling the antibody/antigen binding process in immunoassays. Specifically, more target biomolecules can be attached/tagged and analyzed since the three dimensional structure of the magnetic particles increases the surface to volume ratio. Additionally, such biomolecular-tagged magnetic particles can be easily manipulated by an external magnetic field due to their superparamagnetic behavior. Therefore, magnetic particle- based immunoassays are extensively applied in micro-flow cytometry. The design of a square-loop micro-trap as a magnetic particle manipulator as well as numerical and experimental analysis is presented. Experimental results showed that the micro-trap could successfully trap and concentrate magnetic particles from a large to a small area with a high spatial range.

  15. Long Term RadNet Quality Data

    Data.gov (United States)

    U.S. Environmental Protection Agency — This RadNet Quality Data Asset includes all data since initiation and when ERAMS was expanded to become RadNet, name changed to reflect new mission. This includes...

  16. Introduktion til udvikling af bayesianske net

    DEFF Research Database (Denmark)

    Aagaard, Morten

    2007-01-01

    Bogen introducerer på let og uformelt de centrale grundbegreber i bayesianske net og introducerer til en udviklingsmetode for software, som inkluderer bayesianske net. Metoden kaldes BNAD og inkluderer aktiviteterne preprojekt, kvalitativ modellering, kvantitativ modellering samt...

  17. Special Section on Coloured Petri Nets

    DEFF Research Database (Denmark)

    1998-01-01

    Special section on coloured Petri nets, their basic concepts, analysis methods, tool support and industrial applications.......Special section on coloured Petri nets, their basic concepts, analysis methods, tool support and industrial applications....

  18. RadNet Databases and Reports

    Science.gov (United States)

    EPA’s RadNet data are available for viewing in a searchable database or as PDF reports. Historical and current RadNet monitoring data are used to estimate long-term trends in environmental radiation levels.

  19. Self-assisted optothermal trapping of gold nanorods under two-photon excitation

    Science.gov (United States)

    Chen, Hongtao; Gratton, Enrico; Digman, Michelle A.

    2016-09-01

    We report a self-assisted optothermal trapping and patterning of gold nanorods (GNRs) on glass surfaces with a femtosecond laser. We show that GNRs are not only the trapping targets, but also can enhance the optothermal trapping of other particles. This trapping phenomenon is the net result of thermophoresis and a convective flow caused by localized heating. The heating is due to the conversion of absorbed photons into heat at GNR’s longitudinal surface plasmon resonance (LSPR) wavelength. First, we investigated the optothermal trapping of GNRs at their LSPR wavelength on the glass surface with as low as 0.5 mW laser power. The trapping range was observed to be larger than a typical field of view, e.g. 210 µm  ×  210 µm here. Second, by adjusting the distance between the laser focus and the glass surface, ring patterns of GNRs on the glass surface were obtained. These patterns could be controlled by the laser power and the numerical aperture of the microscope objective. Moreover, we examined the spectral emission of GNRs under different trapping conditions using the spectral phasor approach to reveal the temperature and association status of GNRs. Our study will help understanding manipulation of flows in solution and in biological systems that can be applied in future investigations of GNR-induced heating and flows.

  20. Human Leukocytes Kill Brugia malayi Microfilariae Independently of DNA-Based Extracellular Trap Release

    OpenAIRE

    Ciaran J. McCoy; Reaves, Barbara J.; Giguère, Steeve; Coates, Ruby; Rada, Balázs; Wolstenholme, Adrian J.

    2017-01-01

    Background Wuchereria bancrofti, Brugia malayi and Brugia timori infect over 100 million people worldwide and are the causative agents of lymphatic filariasis. Some parasite carriers are amicrofilaremic whilst others facilitate mosquito-based disease transmission through blood-circulating microfilariae (Mf). Recent findings, obtained largely from animal model systems, suggest that polymorphonuclear leukocytes (PMNs) contribute to parasitic nematode-directed type 2 immune responses. When expos...

  1. Do Size and Insecticide Treatment Matter? Evaluation of Different Nets against Phlebotomus argentipes, the Vector of Visceral Leishmaniasis in Nepal.

    Directory of Open Access Journals (Sweden)

    Murari Lal Das

    Full Text Available In the Indian subcontinent, Leishmania donovani, the parasite causing visceral leishmaniasis (VL is transmitted by the sand fly vector Phlebotomus argentipes. Long lasting insecticide treated nets (LN have been postulated as alternative or complement to Indoor Residual Spraying but there are few field studies evaluating the entomological efficacy of different nets against this vector. We conducted two crossover trials in a VL endemic area in Nepal to compare the barrier effect of (1 LN with different mesh sizes (156 holes/inch2 vs 625 holes/inch2 and (2 alpha-cypermethrin treated LN and untreated nets having the same mesh size (156 holes/inch2. Each crossover trial had two arms consisting of a sequence of two different nets for 8 nights. We used 10 cattle sheds per trial. A cow placed under the net was used as bait. CDC light traps placed inside the nets were used to evaluate the number of P. argentipes crossing the net barrier. Negative binomial generalized estimating equation (GEE population-averaged models adjusted by night and sequence were used to estimate the barrier effect of the different nets. The crossover trials conducted in a rural village in Morang district (South-eastern Nepal demonstrated that reducing the size of the holes in treated nets (625 holes/inch2 increased the barrier effect of LN by 77% (95% confidence interval (CI: 56%-88% compared with treated nets with larger holes (156 holes/inch2. Treating nets with alpha-cypermethrin reduced the number of P. argentipes captured inside the nets by 77% (95% CI: 27%-93% compared with untreated nets. The effectiveness and acceptability of finer mesh pyrethroid treated LN should be tested for VL prevention in a randomized controlled trial.

  2. Do Size and Insecticide Treatment Matter? Evaluation of Different Nets against Phlebotomus argentipes, the Vector of Visceral Leishmaniasis in Nepal

    Science.gov (United States)

    Das, Murari Lal; Rowland, Mark; Austin, James W.; De Lazzari, Elisa; Picado, Albert

    2014-01-01

    In the Indian subcontinent, Leishmania donovani, the parasite causing visceral leishmaniasis (VL) is transmitted by the sand fly vector Phlebotomus argentipes. Long lasting insecticide treated nets (LN) have been postulated as alternative or complement to Indoor Residual Spraying but there are few field studies evaluating the entomological efficacy of different nets against this vector. We conducted two crossover trials in a VL endemic area in Nepal to compare the barrier effect of (1) LN with different mesh sizes (156 holes/inch2 vs 625 holes/inch2) and (2) alpha-cypermethrin treated LN and untreated nets having the same mesh size (156 holes/inch2). Each crossover trial had two arms consisting of a sequence of two different nets for 8 nights. We used 10 cattle sheds per trial. A cow placed under the net was used as bait. CDC light traps placed inside the nets were used to evaluate the number of P. argentipes crossing the net barrier. Negative binomial generalized estimating equation (GEE) population-averaged models adjusted by night and sequence were used to estimate the barrier effect of the different nets. The crossover trials conducted in a rural village in Morang district (South-eastern Nepal) demonstrated that reducing the size of the holes in treated nets (625 holes/inch2) increased the barrier effect of LN by 77% (95% confidence interval (CI): 56%–88%) compared with treated nets with larger holes (156 holes/inch2). Treating nets with alpha-cypermethrin reduced the number of P. argentipes captured inside the nets by 77% (95% CI: 27%–93%) compared with untreated nets. The effectiveness and acceptability of finer mesh pyrethroid treated LN should be tested for VL prevention in a randomized controlled trial. PMID:25494099

  3. Trap induction and trapping in eight nematode-trapping fungi (Orbiliaceae) as affected by juvenile stage of Caenorhabditis elegans.

    Science.gov (United States)

    Xie, Hongyan; Aminuzzaman, F M; Xu, Lingling; Lai, Yiling; Li, Feng; Liu, Xingzhong

    2010-06-01

    This study measured trap induction and trapping on agar disks as affected by juvenile stages (J1, J2, J3, and J4) of the nematode Caenorhabditis elegans and by species of nematode-trapping fungi. Eight species of nematode-trapping fungi belonging to the family Orbiliaceae and producing four kinds of traps were studied: adhesive network-forming Arthrobotrys oligospora, A. vermicola, and A. eudermata, constricting ring-forming Drechslerella brochopaga, and Dr. stenobrocha, adhesive column-forming Dactylellina cionopaga, and adhesive knob-forming Da. ellipsospora, and Da. drechsleri. The number of traps induced generally increased with increasing juvenile stages of C. elegans. The ability to capture the juveniles tended to be similar among isolates that produced the same kind of trap but differed among species that produced different kinds of traps. Trapping by Dr. stenobrocha and Da. cionopaga was correlated with trap number and with juvenile stage. A. oligospora and A. vermicola respectively captured more than 92 and 88% of the J1, J3, and J4 but captured a lower percentage of J2. The knob-producing isolates captured more younger than elder juveniles. Partial correlation analyses demonstrated that the trap induction of the most fungal species positively correlated with the juvenile size and motility, which was juvenile stage dependent. Overall, trap induction and trapping correlated with C. elegans juvenile stage (size and motility) in six species of trapping fungi.

  4. NetBeans IDE 8 cookbook

    CERN Document Server

    Salter, David

    2014-01-01

    If you're a Java developer of any level using NetBeans and want to learn how to get the most out of NetBeans, then this book is for you. Learning how to utilize NetBeans will provide a firm foundation for your Java application development.

  5. Net analyte signal calculation for multivariate calibration

    NARCIS (Netherlands)

    Ferre, J.; Faber, N.M.

    2003-01-01

    A unifying framework for calibration and prediction in multivariate calibration is shown based on the concept of the net analyte signal (NAS). From this perspective, the calibration step can be regarded as the calculation of a net sensitivity vector, whose length is the amount of net signal when the

  6. 78 FR 72451 - Net Investment Income Tax

    Science.gov (United States)

    2013-12-02

    ... Revenue Service 26 CFR Part 1 RIN 1545-BL74 Net Investment Income Tax AGENCY: Internal Revenue Service...). These regulations provide guidance on the computation of net investment income. The regulations affect... lesser of: (A) The individual's net investment income for such taxable year, or (B) the excess (if...

  7. 47 CFR 69.302 - Net investment.

    Science.gov (United States)

    2010-10-01

    ... 47 Telecommunication 3 2010-10-01 2010-10-01 false Net investment. 69.302 Section 69.302... Apportionment of Net Investment § 69.302 Net investment. (a) Investment in Accounts 2001, 1220 and Class B Rural...) Investment in Accounts 2002, 2003 and to the extent such inclusions are allowed by this Commission,...

  8. No such thing as the Net Generation

    NARCIS (Netherlands)

    Dr. H.M.M. van Vliet

    2009-01-01

    The propagandization of a Net Generation adds nothing to our understanding of the digital behaviour of young people. Indeed, it is becoming increasingly obvious that the whole concept of a Net Generation rests on incorrect assumptions. Hence, arguments based on a Net Generation are not only irreleva

  9. 10 CFR 436.20 - Net savings.

    Science.gov (United States)

    2010-01-01

    ... Life Cycle Cost Analyses § 436.20 Net savings. For a retrofit project, net savings may be found by subtracting life cycle costs based on the proposed project from life cycle costs based on not having it. For a new building design, net savings is the difference between the life cycle costs of an...

  10. Nets, tiles, and metal-organic frameworks

    Directory of Open Access Journals (Sweden)

    Michael O’Keeffe

    2014-12-01

    Full Text Available An account is given of the basic nets that are important in the description and design of metal-organic framework (MOF structures. These are generally of minimal transitivity, a concept which is explained. Derived nets are defined and the advantages of using derived nets to describe the topology of MOF frameworks with multiple branch points are emphasized.

  11. The phenomenology of trapped inflation

    CERN Document Server

    Pearce, Lauren; Sorbo, Lorenzo

    2016-01-01

    Trapped inflation is a mechanism in which particle production from the moving inflaton is the main source of friction in the inflaton equation of motion. The produced fields source inflaton perturbations, which dominate over the vacuum ones. We employ the set of equations for the inflaton zero mode and its perturbations which was developed in the original work on trapped inflation, and which we extend to second order in the perturbations. We build on this study by updating the experimental constraints, and by replacing the existing approximate solutions with more accurate ones. We obtain a different numerical value for the amplitude of the scalar power spectrum, and a parametrically different result for the bispectrum. This has implications for the allowed region of parameter space in models of trapped inflation, and for some of the phenomenological results obtained in this region. The main results in the allowed region are the following: monomial inflaton potentials, such as $V \\propto \\varphi,\\, \\varphi^2$ ...

  12. Promoter trapping in Magnaporthe grisea

    Institute of Scientific and Technical Information of China (English)

    LIU Xiao-hong; LU Jian-ping; WANG Jiao-yu; MIN Hang; LIN Fu-cheng

    2006-01-01

    Application of promoter trapping based on transformation in Magnaporthe grisea is reported in this paper. Two promoter-trapping vectors, designated as pCBGFP and pEGFPHPH, were constructed and transformed into protoplasts of M.grisea. A library of 1077 transformants resistant to hygromycin B was generated. Of which, 448 transformants were found to express eGFP gene in different structures ofM. grisea. Three transformants grew slowly, 5 transformants decreased in conidiafion and 7 transformants reduced in pathogenicity greatly among these 448 transformants. Eleven transformants were checked by genomic southern blot randomly, and 9 of which were single-copy insertions. The promoter trapping technique has been applied successfully in M. grisea and can be used as a tool for functional genomic analysis.

  13. Accretion discs trapped near corotation

    OpenAIRE

    D'Angelo, C.R.; Spruit, H.C.

    2012-01-01

    We show that discs accreting on to the magnetosphere of a rotating star can end up in a trapped state, in which the inner edge of the disc stays near the corotation radius, even at low and varying accretion rates. The accretion in these trapped states can be steady or cyclic; we explore these states over a wide range of parameter space. We find two distinct regions of instability: one related to the buildup and release of mass in the disc outside corotation, and the other to mass storage with...

  14. Dysprosium magneto-optical traps

    CERN Document Server

    Youn, Seo Ho; Ray, Ushnish; Lev, Benjamin L

    2010-01-01

    Magneto-optical traps (MOTs) of highly magnetic lanthanides open the door to explorations of novel phases of strongly correlated matter such as lattice supersolids and quantum liquid crystals. We recently reported the first MOTs of the five high abundance isotopes of the most magnetic atom, dysprosium. Described here are details of the experimental technique employed for repumper-free Dy MOTs containing up to half a billion atoms. Extensive characterization of the MOTs' properties---population, temperature, loading, metastable decay dynamics, trap dynamics---is provided.

  15. Feedback traps for virtual potentials

    Science.gov (United States)

    Gavrilov, Momčilo; Bechhoefer, John

    2017-03-01

    Feedback traps are tools for trapping and manipulating single charged objects, such as molecules in solution. An alternative to optical tweezers and other single-molecule techniques, they use feedback to counteract the Brownian motion of a molecule of interest. The trap first acquires information about a molecule's position and then applies an electric feedback force to move the molecule. Since electric forces are stronger than optical forces at small scales, feedback traps are the best way to trap single molecules without `touching' them (e.g. by putting them in a small box or attaching them to a tether). Feedback traps can do more than trap molecules: they can also subject a target object to forces that are calculated to be the gradient of a desired potential function U(x). If the feedback loop is fast enough, it creates a virtual potential whose dynamics will be very close to those of a particle in an actual potential U(x). But because the dynamics are entirely a result of the feedback loop-absent the feedback, there is only an object diffusing in a fluid-we are free to specify and then manipulate in time an arbitrary potential U(x,t). Here, we review recent applications of feedback traps to studies on the fundamental connections between information and thermodynamics, a topic where feedback plays an even more fundamental role. We discuss how recursive maximum-likelihood techniques allow continuous calibration, to compensate for drifts in experiments that last for days. We consider ways to estimate work and heat, using them to measure fluctuating energies to a precision of ±0.03 kT over these long experiments. Finally, we compare work and heat measurements of the costs of information erasure, the Landauer limit of kT ln 2 per bit of information erased. We argue that, when you want to know the average heat transferred to a bath in a long protocol, you should measure instead the average work and then infer the heat using the first law of thermodynamics. This

  16. Rotation sensing with trapped ions

    CERN Document Server

    Campbell, W C

    2016-01-01

    We present a protocol for using trapped ions to measure rotations via matter-wave Sagnac interferometry. The trap allows the interferometer to enclose a large area in a compact apparatus through repeated round-trips in a Sagnac geometry. We show how a uniform magnetic field can be used to close the interferometer over a large dynamic range in rotation speed and measurement bandwidth without losing contrast. Since this technique does not require the ions to be confined in the Lamb-Dicke regime, thermal states with many phonons should be sufficient for operation.

  17. Extracellular DNA in oral microbial biofilms.

    Science.gov (United States)

    Jakubovics, Nicholas S; Burgess, J Grant

    2015-07-01

    The extracellular matrix of microbial biofilms is critical for surface adhesion and nutrient homeostasis. Evidence is accumulating that extracellular DNA plays a number of important roles in biofilm integrity and formation on hard and soft tissues in the oral cavity. Here, we summarise recent developments in the field and consider the potential of targeting DNA for oral biofilm control.

  18. NET 40 Generics Beginner's Guide

    CERN Document Server

    Mukherjee, Sudipta

    2012-01-01

    This is a concise, practical guide that will help you learn Generics in .NET, with lots of real world and fun-to-build examples and clear explanations. It is packed with screenshots to aid your understanding of the process. This book is aimed at beginners in Generics. It assumes some working knowledge of C# , but it isn't mandatory. The following would get the most use out of the book: Newbie C# developers struggling with Generics. Experienced C++ and Java Programmers who are migrating to C# and looking for an alternative to other generic frameworks like STL and JCF would find this book handy.

  19. Complexity Metrics for Workflow Nets

    DEFF Research Database (Denmark)

    Lassen, Kristian Bisgaard; van der Aalst, Wil M.P.

    2009-01-01

    Process modeling languages such as EPCs, BPMN, flow charts, UML activity diagrams, Petri nets, etc.\\ are used to model business processes and to configure process-aware information systems. It is known that users have problems understanding these diagrams. In fact, even process engineers and system......, etc. It seems obvious that the complexity of the model contributes to design errors and a lack of understanding. It is not easy to measure complexity, however. This paper presents three complexity metrics that have been implemented in the process analysis tool ProM. The metrics are defined...

  20. Mapping PUNITY to UniNet

    Institute of Scientific and Technical Information of China (English)

    ZHOU GuoFu(周国富); YUAN ChongYi(袁崇义)

    2003-01-01

    To solve the problems of the interleaving assumption and the single resource inPUNITY (Petri net and UNITY) and Petri net respectively, this paper proposes a set of mappingrules from PUNITY to UniNet. Based on these rules, problems of one field can be transformed toproblems of the other field and powerful tools of Petri net and UNITY can be used. The papergives a sketch of the mapping rules and applies the rules to an example. Meanwhile, the mappingrules can help computer to translate PUNITY to UniNet easily.

  1. Petri nets semantics ofπ-calculus

    Institute of Scientific and Technical Information of China (English)

    Zhenhua YU; Yuanli CAI; Haiping XU

    2008-01-01

    As π-calculus based on the interleaving semantics cannot depict the true concurrency and has few supporting tools,it is translated into Petri nets.π-calculus is divided into basic elements,sequence,concurrency,choice and recursive modules.These modules are translated into Petri nets to construct a complicated system.Petri nets semantics for π-calculus visualize system structure as well as system behaviors.The structural analysis techniques allow direct qualitative analysis of the system properties on the structure of the nets.Finally,Petri nets semantics for π-calculus are illustrated by applying them to mobile telephone systems.

  2. Towards a Standard for Modular Petri Nets

    DEFF Research Database (Denmark)

    Kindler, Ekkart; Petrucci, Laure

    2009-01-01

    as a mechanism for modules in Petri nets that is independent from a particular version of Petri nets and that can mimic many composition mechanisms by a simple import and export concept. Due to its generality, the semantics of modular PNML was only informally defined. Moreover, modular PNML did not define which...... concepts could or should be subject to import and export in high-level Petri nets. In this paper, we formalise a minimal version of modular high-level Petri nets, which is based on the concepts of modular PNML. This shows that modular PNML can be formalised once a specific version of Petri net is fixed...

  3. Delta Semantics Defined By Petri Nets

    DEFF Research Database (Denmark)

    Jensen, Kurt; Kyng, Morten; Madsen, Ole Lehrmann

    This report is identical to an earlier version of May 1978 except that Chapter 5 has been revised. A new paper: "A Petri Net Definition of a System Description Language", DAIMI, April 1979, 20 pages, extends the Petri net model to include a data state representing the program variables. Delta...... and the possibility of using predicates to specify state changes. In this paper a formal semantics for Delta is defined and analysed using Petri nets. Petri nets was chosen because the ideas behind Petri nets and Delta concide on several points. A number of proposals for changes in Delta, which resulted from...

  4. Verifying Time Petri Nets by Linear Programming

    Institute of Scientific and Technical Information of China (English)

    李宣东

    2001-01-01

    The paper proposes an approach to solving some verification problems of time Petri nets using linear programming. The approach is based on the observation that for loop-closed time Petri nets, it is only necessary to investigate a finite prefix of an untimed run of the underlying Petri net. Using the technique the paper gives solutions to reachability and bounded delay timing analysis problems. For both problems algorithms are given, that are decision procedures for loop-closed time Petri nets, and semi-decision procedures for general time Petri nets.

  5. Assessment of a Cross-Vane Trap as a Tool for Sampling the Invasive Megacopta cribraria (Hemiptera: Plataspidae) in Soybean with Associated Evaluations of Female Reproductive Status.

    Science.gov (United States)

    Stubbins, Francesca L; Greene, Jeremy K; Toews, Michael D; Reay-Jones, Francis P F

    2016-10-01

    Megacopta cribraria (F.) (Hemiptera: Plataspidae) is an established pest of soybean, Glycine max (L.) Merrill, in the southeastern United States. Populations of this pest in soybean are typically estimated using sweep nets, a time-consuming procedure. Cross-vane traps may provide an alternative to using sweep nets. The relationship between trap and sweep-net sampling, and the effect of distance from field edges on M. cribraria densities were studied in 2013 and 2014. Adults were detected in the traps up to 3 wk before they were present in the sweep-net samples. Adults and nymphs collected from trap and sweep-net sampling decreased from the field edge toward the field interior. Across all dates and distances, there was a strong positive association between the two sampling methods (r = 0.60), but only 36% of location analyses by week showed significant association. Females collected using both sampling methods were dissected and rated based on egg development. Female populations of all reproductive maturities dispersed into soybean, but proportions varied with sampling method. Results provide a foundation for work into understanding dispersal cues, with implications for management in soybean. Cross-vane traps may serve as an early-season tool to monitor populations, but more studies are needed before robust sampling recommendations can be defined.

  6. Super-Earths as Failed Cores in Orbital Migration Traps

    Science.gov (United States)

    Hasegawa, Yasuhiro

    2016-11-01

    I explore whether close-in super-Earths were formed as rocky bodies that failed to grow fast enough to become the cores of gas giants before the natal protostellar disk dispersed. I model the failed cores’ inward orbital migration in the low-mass or type I regime to stopping points at distances where the tidal interaction with the protostellar disk applies zero net torque. The three kinds of migration traps considered are those due to the dead zone's outer edge, the ice line, and the transition from accretion to starlight as the disk's main heat source. As the disk disperses, the traps move toward final positions near or just outside 1 au. Planets at this location exceeding about 3 M ⊕ open a gap, decouple from their host traps, and migrate inward in the high-mass or type II regime to reach the vicinity of the star. I synthesize the population of planets that formed in this scenario, finding that a fraction of the observed super-Earths could have been failed cores. Most super-Earths that formed this way have more than 4 M ⊕, so their orbits when the disks dispersed were governed by type II migration. These planets have solid cores surrounded by gaseous envelopes. Their subsequent photoevaporative mass loss is most effective for masses originally below about 6 M ⊕. The failed core scenario suggests a division of the observed super-Earth mass-radius diagram into five zones according to the inferred formation history.

  7. Super-Earths as Failed Cores in Orbital Migration Traps

    CERN Document Server

    Hasegawa, Yasuhiro

    2016-01-01

    We explore whether close-in super-Earths were formed as rocky bodies that failed to grow fast enough to become the cores of gas giants before the natal protostellar disk dispersed. We model the failed cores' inward orbital migration in the low-mass or type I regime, to stopping points at distances where the tidal interaction with the protostellar disk applies zero net torque. The three kinds of migration traps considered are those due to the dead zone's outer edge, the ice line, and the transition from accretion to starlight as the disk's main heat source. As the disk disperses, the traps move toward final positions near or just outside 1~au. Planets at this location exceeding about 3~M$_\\oplus$ open a gap, decouple from their host trap, and migrate inward in the high-mass or type II regime to reach the vicinity of the star. We synthesize the population of planets formed in this scenario, finding that some fraction of the observed super-Earths can be failed cores. Most super-Earths formed this way have more t...

  8. Comparison of Bottomless Lift Nets and Breder Traps for Sampling Salt-Marsh Nekton

    Science.gov (United States)

    Vegetated salt-marsh surfaces provide refuge, forage, and spawning habitat for estuarine nekton, yet are threatened by accelerating rates of sea-level rise in southern New England and elsewhere. Nekton responses to ongoing marsh surface changes need to be evaluated with effective...

  9. Microfabricated linear Paul-Straubel ion trap

    Science.gov (United States)

    Mangan, Michael A.; Blain, Matthew G.; Tigges, Chris P.; Linker, Kevin L.

    2011-04-19

    An array of microfabricated linear Paul-Straubel ion traps can be used for mass spectrometric applications. Each ion trap comprises two parallel inner RF electrodes and two parallel outer DC control electrodes symmetric about a central trap axis and suspended over an opening in a substrate. Neighboring ion traps in the array can share a common outer DC control electrode. The ions confined transversely by an RF quadrupole electric field potential well on the ion trap axis. The array can trap a wide array of ions.

  10. Molecular characterization of a new Babesia bovis thrombospondin-related anonymous protein (BbTRAP2.

    Directory of Open Access Journals (Sweden)

    Mohamad Alaa Terkawi

    Full Text Available A gene encoding a Babesia bovis protein that shares significant degree of similarity to other apicomplexan thrombospondin-related anonymous proteins (TRAPs was found in the genomic database and designated as BbTRAP2. Recombinant protein containing a conserved region of BbTRAP2 was produced in E. coli. A high antigenicity of recombinant BbTRAP2 (rBbTRAP2 was observed with field B. bovis-infected bovine sera collected from geographically different regions of the world. Moreover, antiserum against rBbTRAP2 specifically reacted with the authentic protein by Western blot analysis and an indirect fluorescent antibody test. Three bands corresponding to 104-, 76-, and 44-kDa proteins were identified in the parasite lysates and two bands of 76- and 44-kDa proteins were detected in the supernatant of cultivated parasites, indicating that BbTRAP2 was proteolytically processed and shed into the culture. Apical and surface localizations of BbTRAP2 were observed in the intracellular and extracellular parasites, respectively, by confocal laser microscopic examination. Moreover, native BbTRAP2 was precipitated by bovine erythrocytes, suggesting its role in the attachment to erythrocytes. Furthermore, the specific antibody to rBbTRAP2 inhibited the growth of B. bovis in a concentration-dependent manner. Consistently, pre-incubation of the free merozoites with the antibody to rBbTRAP2 resulted in an inhibition of the parasite invasion into host erythrocytes. Interestingly, the antibody to rBbTRAP2 was the most inhibitive for the parasite's growth as compared to those of a set of antisera produced against different recombinant proteins, including merozoite surface antigen 2c (BbMSA-2c, rhoptry-associated protein 1 C-terminal (BbRAP-1CT, and spherical body protein 1 (BbSBP-1. These results suggest that BbTRAP2 might be a potential candidate for development of a subunit vaccine against B. bovis infection.

  11. TRAP1 controls mitochondrial fusion/fission balance through Drp1 and Mff expression.

    Directory of Open Access Journals (Sweden)

    Hironori Takamura

    Full Text Available Mitochondria are dynamic organelles that change in response to extracellular stimuli. These changes are essential for normal mitochondrial/cellular function and are controlled by a tight balance between two antagonistic pathways that promote fusion and fission. Although some molecules have been identified to mediate the mitochondrial fusion and fission process, the underlying mechanisms remain unclear. Tumor necrosis factor receptor-associated protein 1 (TRAP1 is a mitochondrial molecule that regulates a variety of mitochondrial functions. Here, we examined the role of TRAP1 in the regulation of morphology. Stable TRAP1 knockdown cells showed abnormal mitochondrial morphology, and we observed significant decreases in dynamin-related protein 1 (Drp1 and mitochondrial fission factor (Mff, mitochondrial fission proteins. Similar results were obtained by transient knockdown of TRAP1 in two different cell lines, SH-SY5Y neuroblastoma cells and KNS-42 glioma cells. However, TRAP1 knockdown did not affect expression levels of fusion proteins. The reduction in Drp1 and Mff protein levels was rescued following treatment with the proteasome inhibitor MG132. These results suggest that TRAP1 regulates the expression of fission proteins and controls mitochondrial fusion/fission, which affects mitochondrial/cellular function.

  12. Persistent cellular motion control and trapping using mechanotactic signaling.

    Directory of Open Access Journals (Sweden)

    Xiaoying Zhu

    Full Text Available Chemotactic signaling and the associated directed cell migration have been extensively studied owing to their importance in emergent processes of cellular aggregation. In contrast, mechanotactic signaling has been relatively overlooked despite its potential for unique ways to artificially signal cells with the aim to effectively gain control over their motile behavior. The possibility of mimicking cellular mechanotactic signals offers a fascinating novel strategy to achieve targeted cell delivery for in vitro tissue growth if proven to be effective with mammalian cells. Using (i optimal level of extracellular calcium ([Ca(2+]ext = 3 mM we found, (ii controllable fluid shear stress of low magnitude (σ < 0.5 Pa, and (iii the ability to swiftly reverse flow direction (within one second, we are able to successfully signal Dictyostelium discoideum amoebae and trigger migratory responses with heretofore unreported control and precision. Specifically, we are able to systematically determine the mechanical input signal required to achieve any predetermined sequences of steps including straightforward motion, reversal and trapping. The mechanotactic cellular trapping is achieved for the first time and is associated with a stalling frequency of 0.06 ~ 0.1 Hz for a reversing direction mechanostimulus, above which the cells are effectively trapped while maintaining a high level of directional sensing. The value of this frequency is very close to the stalling frequency recently reported for chemotactic cell trapping [Meier B, et al. (2011 Proc Natl Acad Sci USA 108:11417-11422], suggesting that the limiting factor may be the slowness of the internal chemically-based motility apparatus.

  13. Mass Trapping for Anastrepha suspensa

    Science.gov (United States)

    ABSTRACT In field tests conducted in south Florida to test grape juice as an alternative inexpensive bait for Anastrepha suspensa Loew, high numbers of Zaprionus indianus Gupta were captured in traps baited with aqueous grape juice. These experiments included comparisons of grape juice with standard...

  14. Quantum Games in ion traps

    Energy Technology Data Exchange (ETDEWEB)

    Buluta, Iulia Maria [Department of Quantum Engineering and Systems Science, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656 (Japan)]. E-mail: noa@lyman.q.t.u-tokyo.ac.jp; Fujiwara, Shingo [Department of Quantum Engineering and Systems Science, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656 (Japan)]. E-mail: fujiwara@lyman.q.t.u-tokyo.ac.jp; Hasegawa, Shuichi [Department of Quantum Engineering and Systems Science, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656 (Japan)]. E-mail: hasegawa@q.t.u-tokyo.ac.jp

    2006-10-09

    We propose a general, scalable framework for implementing two-choices-multiplayer Quantum Games in ion traps. In particular, we discuss two famous examples: the Quantum Prisoners' Dilemma and the Quantum Minority Game. An analysis of decoherence due to intensity fluctuations in the applied laser fields is also provided.

  15. Optical trapping of gold aerosols

    DEFF Research Database (Denmark)

    Schmitt, Regina K.; Pedersen, Liselotte Jauffred; Taheri, S. M.

    2015-01-01

    Aerosol trapping has proven challenging and was only recently demonstrated.1 This was accomplished by utilizing an air chamber designed to have a minimum of turbulence and a laser beam with a minimum of aberration. Individual gold nano-particles with diameters between 80 nm and 200 nm were trappe...

  16. Optical trapping of coated microspheres

    NARCIS (Netherlands)

    Bormuth, V.; Jannasch, A.; Ander, M.; van Kats, C.M.; van Blaaderen, A.; Howard, J.; Schäffer, E.

    2008-01-01

    In an optical trap, micron-sized dielectric particles are held by a tightly focused laser beam. The optical force on the particle is composed of an attractive gradient force and a destabilizing scattering force. We hypothesized that using anti-reflection-coated microspheres would reduce scattering a

  17. Optical trapping of coated microspheres.

    Science.gov (United States)

    Bormuth, Volker; Jannasch, Anita; Ander, Marcel; van Kats, Carlos M; van Blaaderen, Alfons; Howard, Jonathon; Schäffer, Erik

    2008-09-01

    In an optical trap, micron-sized dielectric particles are held by a tightly focused laser beam. The optical force on the particle is composed of an attractive gradient force and a destabilizing scattering force. We hypothesized that using anti-reflection-coated microspheres would reduce scattering and lead to stronger trapping. We found that homogeneous silica and polystyrene microspheres had a sharp maximum trap stiffness at a diameter of around 800 nm--the trapping laser wavelength in water--and that a silica coating on a polystyrene microsphere was a substantial improvement for larger diameters. In addition, we noticed that homogeneous spheres of a correct size demonstrated anti-reflective properties. Our results quantitatively agreed with Mie scattering calculations and serve as a proof of principle. We used a DNA stretching experiment to confirm the large linear range in detection and force of the coated microspheres and performed a high-force motor protein assay. These measurements show that the surfaces of the coated microspheres are compatible with biophysical assays.

  18. Funnel traps capture a higher proportion of juvenile Great Tits Parus major than automatic traps

    Science.gov (United States)

    Senar, J.C.; Domenech, J.; Conroy, M.J.

    1999-01-01

    We compared capture rates of Great Tits at funnel traps, where several birds can be captured at once so that some decoy effect may appear, to those obtained at automatic traps, where only one bird can be trapped at a time, at trapping stations in northeastern Spain. Juvenile birds were mainly captured at funnel traps (79% of juvenile captures), whereas adult plumaged birds were captured at both types of traps (51% of captures were at the funnel traps) (test between ages, PTits had lower body condition as measured by ptilochronology (P<0.01). These birds are more easily trapped in funnel traps, which may be acting as decoy traps, and thus are vulnerable to the same kinds of biases (eg age or body condition) that have been previously documented for decoy traps.

  19. 50 CFR 697.19 - Trap limits and trap tag requirements for vessels fishing with lobster traps.

    Science.gov (United States)

    2010-10-01

    ... vessels fishing with lobster traps. 697.19 Section 697.19 Wildlife and Fisheries FISHERY CONSERVATION AND... requirements for vessels fishing with lobster traps. (a) Trap limits for vessels fishing or authorized to fish... management area designation certificate or valid limited access American lobster permit specifying one...

  20. Extracellular DNA metabolism in Haloferax volcanii

    Directory of Open Access Journals (Sweden)

    Scott eChimileski

    2014-02-01

    Full Text Available Extracellular DNA is found in all environments and is a dynamic component of the micro-bial ecosystem. Microbial cells produce and interact with extracellular DNA through many endogenous mechanisms. Extracellular DNA is processed and internalized for use as genetic information and as a major source of macronutrients, and plays several key roles within prokaryotic biofilms. Hypersaline sites contain some of the highest extracellular DNA con-centrations measured in nature–a potential rich source of carbon, nitrogen and phosphorus for halophilic microorganisms. We conducted DNA growth studies for the halophilic archaeon Haloferax volcanii DS2 and show that this model Halobacteriales strain is capable of using exogenous double-stranded DNA as a nutrient. Further experiments with varying medium composition, DNA concentration and DNA types revealed that DNA is utilized primarily as a phosphorus source, that growth on DNA is concentration-dependent and that DNA isolated from different sources is metabolized selectively, with a bias against highly divergent methylated DNA sources. Additionally, fluorescence microscopy experiments showed that labeled DNA colocalized with Haloferax volcanii cells. The gene Hvo_1477 was also identified using a comparative genomic approach as a factor likely to be involved in extracellular DNA processing at the cell surface, and deletion of Hvo_1477 created an H. volcanii strain deficient in its ability to grow on extracellular DNA. Widespread distribution of Hvo_1477 homologs in archaea suggests metabolism of extracellular DNA may be of broad ecological and physiological relevance in this domain of life.

  1. The QuarkNet Collaboration

    Science.gov (United States)

    Erzberger, A.

    2003-12-01

    QuarkNet is a long-term high school education project, supported by NSF and DOE and carried out by a collaboration of university and laboratory research groups. These research groups are part of major international particle physics experiments, including those at CERN in Switzerland, Fermilab in Illinois, and SLAC in California. Goals and Objectives: A major goal is to engage students and teachers in authentic scientific research; they gain a first-hand understanding of research and its application in the inquiry method of learning. Teachers enhance their content knowledge, increase their abilities to solve science-related problems, engage students in scientific inquiry, and develop responsibility for their own professional development. Students learn fundamental physics and are motivated by current research questions as they analyze real data. A second goal is to engage particle physicists with current issues in science education, including their understanding of the National Science Education Standards and local science education needs and what constitutes age-appropriate content. Project Design: Working with physicists nationwide, we have established a project framework with three program areas-teacher research experience, teacher development programs, and online resources and inquiry-based activities. Eight-week research appointments allow teachers to experience scientific research first-hand. In teacher institutes the next summer these teachers and scientists lead a group of teachers through a short research scenario lasting two to three weeks and assist them in creating similar scenarios for their students. When fully implemented QuarkNet will support centers associated with 60 particle physics research groups at universities and laboratories in the U. S. The QuarkNet website provides: - Experimental data for use in inquiry-based activities. - Opportunities for communication and collaboration among physicists, teachers and students. - A place for students to

  2. Scaling ion traps for quantum computing

    CSIR Research Space (South Africa)

    Uys, H

    2010-09-01

    Full Text Available The design, fabrication and preliminary testing of a chipscale, multi-zone, surface electrode ion trap is reported. The modular design and fabrication techniques used are anticipated to advance scalability of ion trap quantum computing architectures...

  3. A Portable Root-door Hog Trap

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — Since the early 1960's, over 4,500 wild hogs have been relocated through Florida Game and Fresh Water Fish Commission trapping operations. The trap now used by the...

  4. Trapping Plan: Clarence Cannon National Wildlife Refuge

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — The Clarence Cannon NWR trapping plan outlines trapping areas, species, regulations, equipment, and seasons. This plan will allow harvest of a renewable natural...

  5. Trapping Plan: Necedah National Wildlife Refuge

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — The Necedah NWR trapping plan outlines trapping areas, species, regulations, equipment, and seasons. This plan will allow harvest of a renewable natural resource...

  6. Oxford ion-trap quantum computing project.

    Science.gov (United States)

    Lucas, D M; Donald, C J S; Home, J P; McDonnell, M J; Ramos, A; Stacey, D N; Stacey, J-P; Steane, A M; Webster, S C

    2003-07-15

    We describe recent progress in the development of an ion-trap quantum information processor. We discuss the choice of ion species and describe recent experiments on read-out for a ground-state qubit and photoionization trap loading.

  7. Benton Lake National Wildlife Refuge : Trapping Plan

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — The Benton Lake NWR trapping plan outlines trapping areas, species, regulations, equipment, and seasons. This plan will allow harvest of a renewable natural resource...

  8. Trap split with Laguerre-Gaussian beams

    CERN Document Server

    Kazemi, Seyedeh Hamideh; Mahmoud, Mohammad

    2016-01-01

    The optical trapping techniques have been extensively used in physics, biophysics, micro-chemistry, and micro-mechanics to allow trapping and manipulation of materials ranging from particles, cells, biological substances, and polymers to DNA and RNA molecules. In this Letter, we present a convenient and effective way to generate a novel phenomenon of trapping, named trap split, in a conventional four-level double-$\\Lambda$ atomic system driven by four femtosecond Laguerre-Gaussian laser pulses. We find that trap split can be always achieved when atoms are trapped by such laser pulses, as compared to Gaussian ones. This work would greatly facilitate the trapping and manipulating the particles and generation of trap split. It may also suggest the possibility of extension into new research fields, such as micro-machining and biophysics.

  9. Optical and Magnetic Trapping of Potassium 39

    Science.gov (United States)

    Ensher, Jason; Cornell, Eric; Cataliotti, Francesco; Fort, Chiara; Marin, Francesco; Prevedelli, Marco; Inguscio, Massimo; Ricci, Leonardo; Tino, Guglielmo

    1998-05-01

    We present measurments of optical trapping and cooling and magnetic trapping of ^39K in a double-MOT apparatus. (Optics Lett. 21, 290(1996)) We have measured light-assisted collisional loss rates from our second MOT over a range of trap light intensities. At an intensity of 10 mW/cm^2 we find a loss rate parameter β of 2 x 10-11 cc/s. β increases with trap light intensity and is consistent with the values measured by Williamson and Walker (JOSA B 12, 1393 (1995)). We also present studies of the temperature of atoms in a MOT of ^39K. Under certain conditions of repump light intensity and trap light detuning we measure temperatures nearly as low as the Doppler Limit. Finally, we report on prelimiary results of magnetic trapping in which we have trapped several 10^7 atoms in a quadrupole magnetic trap.

  10. Detection of superoxide production in stimulated and unstimulated living cells using new cyclic nitrone spin traps.

    Science.gov (United States)

    Abbas, Kahina; Hardy, Micael; Poulhès, Florent; Karoui, Hakim; Tordo, Paul; Ouari, Olivier; Peyrot, Fabienne

    2014-06-01

    Reactive oxygen species (ROS), including superoxide anion and hydrogen peroxide (H2O2), have a diverse array of physiological and pathological effects within living cells depending on the extent, timing, and location of their production. For measuring ROS production in cells, the ESR spin trapping technique using cyclic nitrones distinguishes itself from other methods by its specificity for superoxide and hydroxyl radical. However, several drawbacks, such as the low spin trapping rate and the spontaneous and cell-enhanced decomposition of the spin adducts to ESR-silent products, limit the application of this method to biological systems. Recently, new cyclic nitrones bearing a triphenylphosphonium (Mito-DIPPMPO) or a permethylated β-cyclodextrin moiety (CD-DIPPMPO) have been synthesized and their spin adducts demonstrated increased stability in buffer. In this study, a comparison of the spin trapping efficiency of these new compounds with commonly used cyclic nitrone spin traps, i.e., 5,5-dimethyl-1-pyrroline N-oxide (DMPO), and analogs BMPO, DEPMPO, and DIPPMPO, was performed on RAW 264.7 macrophages stimulated with phorbol 12-myristate 13-acetate. Our results show that Mito-DIPPMPO and CD-DIPPMPO enable a higher detection of superoxide adduct, with a low (if any) amount of hydroxyl adduct. CD-DIPPMPO, especially, appears to be a superior spin trap for extracellular superoxide detection in living macrophages, allowing measurement of superoxide production in unstimulated cells for the first time. The main rationale put forward for this extreme sensitivity is that the extracellular localization of the spin trap prevents the reduction of the spin adducts by ascorbic acid and glutathione within cells.

  11. High concentration effects of neutral-potential-well interface traps on recombination dc current-voltage lineshape in metal-oxide-silicon transistors

    Science.gov (United States)

    Chen, Zuhui; Jie, Bin B.; Sah, Chih-Tang

    2008-11-01

    Steady-state Shockley-Read-Hall kinetics is employed to explore the high concentration effect of neutral-potential-well interface traps on the electron-hole recombination direct-current current-voltage (R-DCIV) properties in metal-oxide-silicon field-effect transistors. Extensive calculations include device parameter variations in neutral-trapping-potential-well electron interface-trap density NET (charge states 0 and -1), dopant impurity concentration PIM, oxide thickness Xox, forward source/drain junction bias VPN, and transistor temperature T. It shows significant distortion of the R-DCIV lineshape by the high concentrations of the interface traps. The result suggests that the lineshape distortion observed in past experiments, previously attributed to spatial variation in surface impurity concentration and energy distribution of interface traps in the silicon energy gap, can also arise from interface-trap concentration along surface channel region.

  12. Dynamic array of dark optical traps

    DEFF Research Database (Denmark)

    Daria, V.R.; Rodrigo, P.J.; Glückstad, J.

    2004-01-01

    A dynamic array of dark optical traps is generated for simultaneous trapping and arbitrary manipulation of multiple low-index microstructures. The dynamic intensity patterns forming the dark optical trap arrays are generated using a nearly loss-less phase-to-intensity conversion of a phase-encode...... optical traps for simultaneous manipulation of hollow "air-filled" glass microspheres suspended in an aqueous medium. (C) 2004 American Institute of Physics....

  13. Bound states in string nets

    Science.gov (United States)

    Schulz, Marc Daniel; Dusuel, Sébastien; Vidal, Julien

    2016-11-01

    We discuss the emergence of bound states in the low-energy spectrum of the string-net Hamiltonian in the presence of a string tension. In the ladder geometry, we show that a single bound state arises either for a finite tension or in the zero-tension limit depending on the theory considered. In the latter case, we perturbatively compute the binding energy as a function of the total quantum dimension. We also address this issue in the honeycomb lattice where the number of bound states in the topological phase depends on the total quantum dimension. Finally, the internal structure of these bound states is analyzed in the zero-tension limit.

  14. Bound states in string nets

    CERN Document Server

    Schulz, M D; Vidal, J

    2016-01-01

    We discuss the emergence of bound states in the low-energy spectrum of the string-net Hamiltonian in the presence of a string tension. In the ladder geometry, we show that a single bound state arises either for a finite tension or in the zero-tension limit depending on the theory considered. In the latter case, we perturbatively compute the binding energy as a function of the total quantum dimension. We also address this issue in the honeycomb lattice where the number of bound states in the topological phase depends on the total quantum dimension. Finally, the internal structure of these bound states is analyzed in the zero-tension limit.

  15. -Net Approach to Sensor -Coverage

    Directory of Open Access Journals (Sweden)

    Fusco Giordano

    2010-01-01

    Full Text Available Wireless sensors rely on battery power, and in many applications it is difficult or prohibitive to replace them. Hence, in order to prolongate the system's lifetime, some sensors can be kept inactive while others perform all the tasks. In this paper, we study the -coverage problem of activating the minimum number of sensors to ensure that every point in the area is covered by at least sensors. This ensures higher fault tolerance, robustness, and improves many operations, among which position detection and intrusion detection. The -coverage problem is trivially NP-complete, and hence we can only provide approximation algorithms. In this paper, we present an algorithm based on an extension of the classical -net technique. This method gives an -approximation, where is the number of sensors in an optimal solution. We do not make any particular assumption on the shape of the areas covered by each sensor, besides that they must be closed, connected, and without holes.

  16. NETS - Danish participation. Final report

    Energy Technology Data Exchange (ETDEWEB)

    Alsen, S. (Grontmij - Carl Bro, Glostrup (Denmark)); Theel, C. (Baltic Sea Solutions, Holeby (Denmark))

    2008-12-15

    Within the NICe-funded project 'Nordic Environmental Technology Solutions (NETS)' a new type of networking at the Nordic level was organized in order to jointly exploit the rapidly growing market potential in the environmental technology sector. The project aimed at increased and professionalized commercialization of Nordic Cleantech in energy and water business segments through 1) closer cooperation and joint marketing activities, 2) a website, 3) cleantech product information via brochures and publications 4) and participating in relevant trade fairs and other industry events. Facilitating business-to-business activities was another core task for the NETS project partners from Norway, Sweden, Finland and Denmark with the aim to encourage total solutions for combined Cleantech system offers. The project has achieved to establish a Cleantech register of 600 Nordic Cleantech companies, a network of 86 member enterprises, produced several publications and brochures for direct technology promotion and a website for direct access to company profiles and contact data. The project partners have attended 14 relevant international Cleantech trade fairs and conferences and facilitated business-to-business contacts added by capacity building offers through two company workshops. The future challenge for the project partners and Nordic Cleantech will be to coordinate the numerous efforts within the Nordic countries in order to reach concerted action and binding of member companies for reliable services, an improved visibility and knowledge exchange. With Cleantech's growing market influence and public awareness, the need to develop total solutions is increasing likewise. Marketing efforts should be encouraged cross-sectional and cross-border among the various levels of involved actors from both the public and the private sector. (au)

  17. The Aarhus Ion Micro-Trap Project

    DEFF Research Database (Denmark)

    Miroshnychenko, Yevhen; Nielsen, Otto; Poulsen, Gregers

    and installed in an ultra high vacuum chamber, which includes an ablation oven for all-optical loading of the trap [2]. The next steps on the project are to demonstrate the operation of the micro-trap and the cooling of ions using fiber delivered light. [1] D. Grant, Development of Micro-Scale Ion traps, Master...

  18. The Nanocluster Trap endstation at BESSY II

    Directory of Open Access Journals (Sweden)

    Tobias Lau

    2017-05-01

    Full Text Available The Nanocluster Trap endstation at BESSY II combines a cryogenic linear radio-frequency ion trap with an applied magnetic field for x-ray magnetic circular dichroism studies of cold and size-selected trapped ions. Applications include atomic, molecular, and cluster ions as well as ionic complexes.

  19. Superdense Coding via Hot Trapped Ions

    Institute of Scientific and Technical Information of China (English)

    QIN Tao; FENG Mang; GAO Ke-Lin

    2004-01-01

    Superdense coding plays an important role in quantum information and can be performed with trapped ions. By confining the ions in a linear trap or a trap-cavity setup, we propose schemes to implement a reliable superdense coding by means of bichromatic radiation method. Experimental feasibility and reliability for achieving our schemes is discussed in detail.

  20. Microchip-Based Trapped-Atom Clocks

    CERN Document Server

    Vuletic, Vladan; Schleier-Smith, Monika H

    2011-01-01

    This is a chapter of a recently published book entitled Atom Chips, edited by Jakob Reichel and Vladan Vuletic. The contents of this chapter include: Basic Principles; Atomic-Fountain versus Trapped-Atom Clocks; Optical-Transition Clocks versus Microwave Clocks; Clocks with Magnetically Trapped Atoms--Fundamental Limits and Experimental Demonstrations; Readout in Trapped-Atom Clocks; and Spin Squeezing.

  1. Petri Nets for Biologically Motivated Computing

    Directory of Open Access Journals (Sweden)

    J. Kleijn

    2011-01-01

    Full Text Available Petri nets are a general and well-established model of concurrent and distributed computation and behaviour, including that taking place in biological systems. In this survey paper, we are concerned with intrinsic relationships between Petri nets and two formal models inspired by aspects of the functioning of the living cell: membrane systems and reaction systems. In particular, we are interested in the benefits that can result from establishing strong semantical links between Petri nets and membrane systems and reaction systems. We first discuss Petri nets with localities reflecting the compartmentalisation modelled in membrane systems. Then special attention is given to set-nets, a new Petri net model for reaction systems and their qualitative approach to the investigation of the processes carried out by biochemical reactions taking place in the living cell.

  2. and extracellular laccase isoenzymes from Pleurotus ostreatus ...

    African Journals Online (AJOL)

    ZMG

    Colonia Vicentina, Delegación Iztapalapa, 09340 México D.F., México. ... In this study, extracellular laccase enzymes produced by Pleurotus ostreatus was identified in .... the intracellular forms), through the modified zymography method of.

  3. Alternative methods for characterization of extracellular vesicles

    Directory of Open Access Journals (Sweden)

    Fatemeh eMomen-Heravi

    2012-09-01

    Full Text Available Extracellular vesicles are nano-sized vesicles released by all cells in vitro as well as in vivo. Their role has been implicated mainly in cell-cell communication, but also in disease biomarkers and more recently in gene delivery. They represent a snapshot of the cell status at the moment of release and carry bioreactive macromolecules such as nucleic acids, proteins and lipids. A major limitation in this emerging new field is the availability/awareness of techniques to isolate and properly characterize Extracellular vesicles. The lack of gold standards makes comparing different studies very difficult and may potentially hinder some Extracellular vesicles -specific evidence. Characterization of Extracellular vesicles has also recently seen many advances with the use of Nanoparticle Tracking Analysis (NTA, flow cytometry, cryo-EM instruments and proteomic technologies. In this review, we discuss the latest developments in translational technologies involving characterization methods including the facts in their support and the challenges they face.

  4. Factors associated with mosquito net use by individuals in households owning nets in Ethiopia

    Directory of Open Access Journals (Sweden)

    Graves Patricia M

    2011-12-01

    Full Text Available Abstract Background Ownership of insecticidal mosquito nets has dramatically increased in Ethiopia since 2006, but the proportion of persons with access to such nets who use them has declined. It is important to understand individual level net use factors in the context of the home to modify programmes so as to maximize net use. Methods Generalized linear latent and mixed models (GLLAMM were used to investigate net use using individual level data from people living in net-owning households from two surveys in Ethiopia: baseline 2006 included 12,678 individuals from 2,468 households and a sub-sample of the Malaria Indicator Survey (MIS in 2007 included 14,663 individuals from 3,353 households. Individual factors (age, sex, pregnancy; net factors (condition, age, net density; household factors (number of rooms [2006] or sleeping spaces [2007], IRS, women's knowledge and school attendance [2007 only], wealth, altitude; and cluster level factors (rural or urban were investigated in univariate and multi-variable models for each survey. Results In 2006, increased net use was associated with: age 25-49 years (adjusted (a OR = 1.4, 95% confidence interval (CI 1.2-1.7 compared to children U5; female gender (aOR = 1.4; 95% CI 1.2-1.5; fewer nets with holes (Ptrend = 0.002; and increasing net density (Ptrend [all nets in HH good] = 1.6; 95% CI 1.2-2.1; increasing net density (Ptrend [per additional space] = 0.6, 95% CI 0.5-0.7; more old nets (aOR [all nets in HH older than 12 months] = 0.5; 95% CI 0.3-0.7; and increasing household altitude (Ptrend Conclusion In both surveys, net use was more likely by women, if nets had fewer holes and were at higher net per person density within households. School-age children and young adults were much less likely to use a net. Increasing availability of nets within households (i.e. increasing net density, and improving net condition while focusing on education and promotion of net use, especially in school-age children

  5. Scalar Trapping and Saxion Cosmology

    CERN Document Server

    Moroi, Takeo; Nakayama, Kazunori; Takimoto, Masahiro

    2013-01-01

    We study in detail the dynamics of a scalar field in thermal bath with symmetry breaking potential. In particular, we focus on the process of trapping of a scalar field at an enhanced symmetry point through the thermal/non-thermal particle production, taking into account the interactions of produced particles with the standard model particles. As an explicit example, we revisit the saxion dynamics with an initial amplitude much larger than the Peccei-Quinn scale and show that the saxion trapping phenomenon happens for the most cases and it often leads to thermal inflation. We also study the saxion dynamics after thermal inflation, and it is shown that thermal dissipation effect on the saxion can relax the axion overproduction problem from the saxion decay.

  6. Fundamental physics in particle traps

    CERN Document Server

    Vogel, Manuel

    2014-01-01

    This volume provides detailed insight into the field of precision spectroscopy and fundamental physics with particles confined in traps. It comprises experiments with electrons and positrons, protons and antiprotons, antimatter and highly charged ions, together with corresponding theoretical background. Such investigations represent stringent tests of quantum electrodynamics and the Standard model, antiparticle and antimatter research, test of fundamental symmetries, constants, and their possible variations with time and space. They are key to various aspects within metrology such as mass measurements and time standards, as well as promising to further developments in quantum information processing. The reader obtains a valuable source of information suited for beginners and experts with an interest in fundamental studies using particle traps.

  7. Tachyon Physics with Trapped Ions

    CERN Document Server

    Lee, Tony E; Cheng, Xiao-Hang; Lamata, Lucas; Solano, Enrique

    2015-01-01

    It has been predicted that particles with imaginary mass, called tachyons, would be able to travel faster than the speed of light. So far, there has not been any experimental evidence for tachyons in either natural or engineered systems. Here, we propose how to experimentally simulate Dirac tachyons with trapped ions. Quantum measurement on a Dirac particle simulated by a trapped ion causes it to have an imaginary mass so that it may travel faster than the effective speed of light. We show that a Dirac tachyon must have spinor-motion entanglement in order to be superluminal. We also show that it exhibits significantly more Klein tunneling than a normal Dirac particle. We provide numerical simulations with realistic ion systems and show that our scheme is feasible with current technology.

  8. Bacterial binding to extracellular proteins - in vitro adhesion

    DEFF Research Database (Denmark)

    Schou, C.; Fiehn, N.-E.

    1999-01-01

    Viridans streptococci, bacterial adherence, extracellular matrix proteins, surface receptors, endocarditis......Viridans streptococci, bacterial adherence, extracellular matrix proteins, surface receptors, endocarditis...

  9. Evaluating a portable cylindrical bait trap to capture diamondback terrapins in salt marsh

    Science.gov (United States)

    Henry, Paula F.; Haramis, Michael; Day, Daniel

    2016-01-01

    Diamondback terrapins (Malaclemys terrapin) are currently in decline across much of their historical range, and demographic data on a regional scale are needed to identify where their populations are at greatest risk. Because terrapins residing in salt marshes are difficult to capture, we designed a cylindrical bait trap (CBT) that could be deployed in shallow tidal waters. From 2003 to 2006, trials were conducted with CBTs in the Chesapeake Bay, Maryland (USA) to determine terrapin sex, size, and age distribution within 3 salt marsh interior habitats—open bays, tidal guts, and broken marshes—using 15 traps/habitat. Analyses based on 791 total captures with CBTs indicate that smaller terrapins, (i.e., adult male and subadult) were more prevalent within the transecting tidal guts and broken marshes, whereas the adult females were more evenly distributed among habitats, including open bays. Subadult females made up the largest percent of catch in the CBTs deployed within the 3 marsh interior habitats. During a 12-day trial in which we compared capture performance of CBTs and modified fyke nets along open shorelines during the nesting season, fyke nets outperformed CBTs by accounting for 95.2% of the 604 terrapin captures. Although the long drift leads of the fyke nets proved more effective for intercepting along-shore travel of adult female terrapins during the nesting season, CBTs provided a more effective means of live-trapping terrapins within the shallow interior marshes.

  10. The extracellular RNA complement of Escherichia coli.

    Science.gov (United States)

    Ghosal, Anubrata; Upadhyaya, Bimal Babu; Fritz, Joëlle V; Heintz-Buschart, Anna; Desai, Mahesh S; Yusuf, Dilmurat; Huang, David; Baumuratov, Aidos; Wang, Kai; Galas, David; Wilmes, Paul

    2015-01-21

    The secretion of biomolecules into the extracellular milieu is a common and well-conserved phenomenon in biology. In bacteria, secreted biomolecules are not only involved in intra-species communication but they also play roles in inter-kingdom exchanges and pathogenicity. To date, released products, such as small molecules, DNA, peptides, and proteins, have been well studied in bacteria. However, the bacterial extracellular RNA complement has so far not been comprehensively characterized. Here, we have analyzed, using a combination of physical characterization and high-throughput sequencing, the extracellular RNA complement of both outer membrane vesicle (OMV)-associated and OMV-free RNA of the enteric Gram-negative model bacterium Escherichia coli K-12 substrain MG1655 and have compared it to its intracellular RNA complement. Our results demonstrate that a large part of the extracellular RNA complement is in the size range between 15 and 40 nucleotides and is derived from specific intracellular RNAs. Furthermore, RNA is associated with OMVs and the relative abundances of RNA biotypes in the intracellular, OMV and OMV-free fractions are distinct. Apart from rRNA fragments, a significant portion of the extracellular RNA complement is composed of specific cleavage products of functionally important structural noncoding RNAs, including tRNAs, 4.5S RNA, 6S RNA, and tmRNA. In addition, the extracellular RNA pool includes RNA biotypes from cryptic prophages, intergenic, and coding regions, of which some are so far uncharacterised, for example, transcripts mapping to the fimA-fimL and ves-spy intergenic regions. Our study provides the first detailed characterization of the extracellular RNA complement of the enteric model bacterium E. coli. Analogous to findings in eukaryotes, our results suggest the selective export of specific RNA biotypes by E. coli, which in turn indicates a potential role for extracellular bacterial RNAs in intercellular communication. © 2015 The

  11. Lift enhancement by trapped vortex

    Science.gov (United States)

    Rossow, Vernon J.

    1992-01-01

    The viewgraphs and discussion of lift enhancement by trapped vortex are provided. Efforts are continuously being made to find simple ways to convert wings of aircraft from an efficient cruise configuration to one that develops the high lift needed during landing and takeoff. The high-lift configurations studied here consist of conventional airfoils with a trapped vortex over the upper surface. The vortex is trapped by one or two vertical fences that serve as barriers to the oncoming stream and as reflection planes for the vortex and the sink that form a separation bubble on top of the airfoil. Since the full three-dimensional unsteady flow problem over the wing of an aircraft is so complicated that it is hard to get an understanding of the principles that govern the vortex trapping process, the analysis is restricted here to the flow field illustrated in the first slide. It is assumed that the flow field between the two end plates approximates a streamwise strip of the flow over a wing. The flow between the endplates and about the airfoil consists of a spanwise vortex located between the suction orifices in the endplates. The spanwise fence or spoiler located near the nose of the airfoil serves to form a separated flow region and a shear layer. The vorticity in the shear layer is concentrated into the vortex by withdrawal of fluid at the suction orifices. As the strength of the vortex increases with time, it eventually dominates the flow in the separated region so that a shear or vertical layer is no longer shed from the tip of the fence. At that point, the vortex strength is fixed and its location is such that all of the velocity contributions at its center sum to zero thereby making it an equilibrium point for the vortex. The results of a theoretical analysis of such an idealized flow field are described.

  12. TRAP SYNDROME: A RARE ENTITY

    OpenAIRE

    Varsha; Kanan; Chanda,, Abhra; Qazi,, S.

    2014-01-01

    TRAP (Twin Reversed Arterial Perfusion) sequence is a rare complication of monozygotic twin pregnancy. It is due to defect in early pregnancy where there is defective arterial anastomosis in placenta. This results in a fetus with an absent heart (acardiac twin). This acardiac twin has a poorly developed upper body and head also. The pump twin (with a normal heart) is also at a risk of heart failure and problems related to preterm birth1.

  13. TRAP SYNDROME: A RARE ENTITY

    Directory of Open Access Journals (Sweden)

    Varsha

    2014-04-01

    Full Text Available TRAP (Twin Reversed Arterial Perfusion sequence is a rare complication of monozygotic twin pregnancy. It is due to defect in early pregnancy where there is defective arterial anastomosis in placenta. This results in a fetus with an absent heart (acardiac twin. This acardiac twin has a poorly developed upper body and head also. The pump twin (with a normal heart is also at a risk of heart failure and problems related to preterm birth1.

  14. Trapper readies trap for lizard

    Science.gov (United States)

    2000-01-01

    State-licensed animal trapper James Dean sets the open door of an animal trap on KSC. He hopes to catch a large monitor lizard spotted recently near S.R. 3, a route into the Center, by several area residents. The lizard is not a native of the area, and possibly a released pet. Dean is working with the cooperation of KSC and the Merritt Island National Wildlife Refuge.

  15. NASA Net Zero Energy Buildings Roadmap

    Energy Technology Data Exchange (ETDEWEB)

    Pless, S.; Scheib, J.; Torcellini, P.; Hendron, B.; Slovensky, M.

    2014-10-01

    In preparation for the time-phased net zero energy requirement for new federal buildings starting in 2020, set forth in Executive Order 13514, NASA requested that the National Renewable Energy Laboratory (NREL) to develop a roadmap for NASA's compliance. NASA detailed a Statement of Work that requested information on strategic, organizational, and tactical aspects of net zero energy buildings. In response, this document presents a high-level approach to net zero energy planning, design, construction, and operations, based on NREL's first-hand experience procuring net zero energy construction, and based on NREL and other industry research on net zero energy feasibility. The strategic approach to net zero energy starts with an interpretation of the executive order language relating to net zero energy. Specifically, this roadmap defines a net zero energy acquisition process as one that sets an aggressive energy use intensity goal for the building in project planning, meets the reduced demand goal through energy efficiency strategies and technologies, then adds renewable energy in a prioritized manner, using building-associated, emission- free sources first, to offset the annual energy use required at the building; the net zero energy process extends through the life of the building, requiring a balance of energy use and production in each calendar year.

  16. Modeling of economic systems with Petri nets

    Directory of Open Access Journals (Sweden)

    Pavel Valeryevich Skorodumov

    2014-09-01

    Full Text Available Modeling is one of the most important tools to study complex systems. Components of both continuous and discrete nature are present in the behavior of contemporary economic systems. The article uses formalism of nested hybrid Petri nets as a tool to study complex economic systems. The author describes basic approaches of simulation modelling, concepts of classical Petri nets, modified means of nested hybrid Petri nets, benefits of their use for systems modeling. The article presents the concept of a universal system of simulation modeling. On the basis of considered approaches the article proposes to develop a universal system of simulation modeling on the basis of the modified machine Petri nets

  17. Pro Agile NET Development with Scrum

    CERN Document Server

    Blankenship, Jerrel; Millett, Scott

    2011-01-01

    Pro Agile .NET Development with SCRUM guides you through a real-world ASP.NET project and shows how agile methodology is put into practice. There is plenty of literature on the theory behind agile methodologies, but no book on the market takes the concepts of agile practices and applies these in a practical manner to an end-to-end ASP.NET project, especially the estimating, requirements and management aspects of a project. Pro Agile .NET Development with SCRUM takes you through the initial stages of a project - gathering requirements and setting up an environment - through to the development a

  18. Professional Visual Basic 2010 and .NET 4

    CERN Document Server

    Sheldon, Bill; Sharkey, Kent

    2010-01-01

    Intermediate and advanced coverage of Visual Basic 2010 and .NET 4 for professional developers. If you've already covered the basics and want to dive deep into VB and .NET topics that professional programmers use most, this is your book. You'll find a quick review of introductory topics-always helpful-before the author team of experts moves you quickly into such topics as data access with ADO.NET, Language Integrated Query (LINQ), security, ASP.NET web programming with Visual Basic, Windows workflow, threading, and more. You'll explore all the new features of Visual Basic 2010 as well as all t

  19. Pro ASP.NET MVC 4

    CERN Document Server

    Freeman, Adam

    2012-01-01

    The ASP.NET MVC 4 Framework is the latest evolution of Microsoft's ASP.NET web platform. It provides a high-productivity programming model that promotes cleaner code architecture, test-driven development, and powerful extensibility, combined with all the benefits of ASP.NET. ASP.NET MVC 4 contains a number of significant advances over previous versions. New mobile and desktop templates (employing adaptive rendering) are included together with support for jQuery Mobile for the first time. New display modes allow your application to select views based on the browser that's making the request whi

  20. Discrete, Continuous, and Hybrid Petri Nets

    CERN Document Server

    David, Rene

    2010-01-01

    Petri Nets were introduced and still successfully used to analyze and model discrete event systems especially in engineering and computer sciences such as in automatic control. Recently this discrete Petri Nets formalism was successfully extended to continuous and hybrid systems. This monograph presents a well written and clearly organized introduction in the standard methods of Petri Nets with the aim to reach an accurate understanding of continuous and hybrid Petri Nets, while preserving the consistency of basic concepts throughout the book. The book is a monograph as well as a didactic tool

  1. Towards a Standard for Modular Petri Nets

    DEFF Research Database (Denmark)

    Kindler, Ekkart; Petrucci, Laure

    2009-01-01

    When designing complex systems, mechanisms for structuring, composing, and reusing system components are crucial. Today, there are many approaches for equipping Petri nets with such mechanisms. In the context of defining a standard interchange format for Petri nets, modular PNML was defined....... Moreover, we present and discuss some more advanced features of modular Petri nets that could be included in the standard. This way, we provide a formal foundation and a basis for a discussion of features to be included in the upcoming standard of a module concept for Petri nets in general and for high...

  2. Extracellular DNA-induced antimicrobial peptide resistance mechanisms in Pseudomonas aeruginosa.

    Directory of Open Access Journals (Sweden)

    Shawn eLewenza

    2013-02-01

    Full Text Available Extracellular DNA (eDNA is in the environment, bodily fluids, in the matrix of biofilms, and accumulates at infection sites. Extracellular DNA can function as a nutrient source, a universal biofilm matrix component and an innate immune effector in extracellular DNA traps. In biofilms, eDNA is required for attachment, aggregation and stabilization of microcolonies. We have recently shown that eDNA can sequester divalent metal cations, which has interesting implications on antibiotic resistance. Extracellular DNA binds metal cations and thus activates the Mg2+-responsive PhoPQ and PmrAB two-component systems. In Pseudomonas aeruginosa and many other Gram-negative bacteria, the PhoPQ/PmrAB systems control various genes required for virulence and resisting killing by antimicrobial peptides, including the pmr genes (PA3552-PA3559 that are responsible for the addition of aminoarabinose to lipid A. The PA4773-PA4775 genes are a second DNA-induced cluster and are required for the production of spermidine on the outer surface, which protects the outer membrane from antimicrobial peptide treatment. Both modifications mask the negative surface charges and limit membrane damage by antimicrobial peptides. DNA-enriched biofilms or planktonic cultures have increased antibiotic resistance phenotypes to antimicrobial peptides and aminoglycosides. These dual antibiotic resistance and immune evasion strategies may be expressed in DNA-rich environments and contribute to long-term survival.

  3. Antiparticle plasmas for antihydrogen trapping

    Science.gov (United States)

    Charlton, M.; Andresen, G. B.; Ashkezari, M. D.; Baquero-Ruiz, M.; Bertsche, W.; Bowe, P. D.; Butler, E.; Carpenter, P. T.; Cesar, C. L.; Chapman, S.; Eriksson, S.; Fajans, J.; Friesen, T.; Fujiwara, M. C.; Gill, D. R.; Gutierrez, A.; Hangst, J. S.; Hardy, W. N.; Hayano, R. S.; Hayden, M. E.; Humphries, A. J.; Hurt, J. L.; Hydomako, R.; Jonsell, S.; Kurchaninov, L.; Madsen, N.; Menary, S.; Nolan, P.; Olchanski, K.; Olin, A.; Povilus, A.; Pusa, P.; Robicheaux, F.; Sarid, E.; Silveira, D. M.; So, C.; Storey, J. W.; Thompson, R. I.; van der Werf, D. P.; Wurtele, J. S.; Yamazaki, Y.

    2012-05-01

    Over the last decades it has become routine to form beams of positrons and antiprotons and to use them to produce trapped samples of both species for a variety of purposes. Positrons can be captured efficiently, for instance using a buffer-gas system, and in such quantities to form dense, single component plasmas useful for antihydrogen formation. The latter is possible using developments of techniques for dynamically capturing and then cooling antiprotons ejected from the Antiproton Decelerator at CERN. The antiprotons can then be manipulated by cloud compression and evaporative cooling to form tailored plasmas. We will review recent advances that have allowed antihydrogen atoms to be confined for the first time in a shallow magnetic minimum neutral atom trap superimposed upon the region in which the antiparticles are held and mixed. A new mixing technique has been developed to help achieve this using autoresonant excitation of the centreofmass longitudinal motion of an antiproton cloud. This allows efficient antihydrogen formation without imparting excess energy to the antiprotons and helps enhance the probability of trapping the anti-atom.

  4. Transcriptome of extracellular vesicles released by hepatocytes.

    Directory of Open Access Journals (Sweden)

    Felix Royo

    Full Text Available The discovery that the cells communicate through emission of vesicles has opened new opportunities for better understanding of physiological and pathological mechanisms. This discovery also provides a novel source for non-invasive disease biomarker research. Our group has previously reported that hepatocytes release extracellular vesicles with protein content reflecting the cell-type of origin. Here, we show that the extracellular vesicles released by hepatocytes also carry RNA. We report the messenger RNA composition of extracellular vesicles released in two non-tumoral hepatic models: primary culture of rat hepatocytes and a progenitor cell line obtained from a mouse foetal liver. We describe different subpopulations of extracellular vesicles with different densities and protein and RNA content. We also show that the RNA cargo of extracellular vesicles released by primary hepatocytes can be transferred to rat liver stellate-like cells and promote their activation. Finally, we provide in vitro and in vivo evidence that liver-damaging drugs galactosamine, acetaminophen, and diclofenac modify the RNA content of these vesicles. To summarize, we show that the extracellular vesicles secreted by hepatocytes contain various RNAs. These vesicles, likely to be involved in the activation of stellate cells, might become a new source for non-invasive identification of the liver toxicity markers.

  5. Hybrid ion, atom and light trap

    CERN Document Server

    Jyothi, S; Ram, N Bhargava; Rangwala, S A

    2013-01-01

    We present an unique experimental arrangement which permits the simultaneous trapping and cooling of ions and neutral atoms, within a Fabry-Perot (FP) cavity. The versatility of this hybrid trap experiment enables a variety of studies with trapped mixtures. The motivations behind the production of such a hybrid trap system are explained, followed by details of how the experiment is put together. Several experiments that have been performed with this system are presented and some opportunities with this system are discussed. However the primary emphasis is focussed on the aspects that pertain to the trapped ions, in this hybrid system.

  6. Collisional blockade in microscopic optical dipole traps.

    Science.gov (United States)

    Schlosser, N; Reymond, G; Grangier, P

    2002-07-08

    We analyze the operating regimes of a very small optical dipole trap, loaded from a magneto-optical trap, as a function of the atom loading rate, i.e., the number of atoms per second entering the dipole trap. We show that, when the dipole trap volume is small enough, a "collisional blockade" mechanism locks the average number of trapped atoms on the value 0.5 over a large range of loading rates. We also discuss the "weak loading" and "strong loading" regimes outside the blockade range, and we demonstrate experimentally the existence of these three regimes.

  7. In-trap conversion electron spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Weissman, L. E-mail: weissman@nscl.msu.edu; Ames, F.; Aeysto, J.; Forstner, O.; Reisinger, K.; Rinta-Antila, S

    2002-10-21

    The Penning trap REXTRAP at ISOLDE was used to test the feasibility of in-trap conversion electron spectroscopy. The results of simulations, experiments with solid conversion electron sources as well as first on-line tests with trapped radioactive ions are presented. In addition to obtaining high-resolution spectroscopic data, the detection of conversion electrons was found to be a useful tool for the diagnostics of the trap operation. The tests proved the feasibility of in-trap spectroscopy but also revealed some potential problems to be addressed in the future.

  8. In-trap conversion electron spectroscopy

    CERN Document Server

    Weissman, L; Äystö, J; Forstner, O; Reisinger, K; Rinta-Antila, S

    2002-01-01

    The Penning trap REXTRAP at ISOLDE was used to test the feasibility of in-trap conversion electron spectroscopy. The results of simulations, experiments with solid conversion electron sources as well as first on-line and tests with trapped radioactive ions are presented. In addition to obtaining high-resolution spectroscopic data, the detection of conversion electrons was found to be a useful tool for the diagnostics of the trap operation. The tests proved the feasibility of in-trap spectroscopy but also revealed some potential problems to be addressed in the future.

  9. Effect of trapped charge accumulation on the retention of charge trapping memory

    Energy Technology Data Exchange (ETDEWEB)

    Jin Rui; Liu Xiaoyan; Du Gang; Kang Jinfeng; Han Ruqi, E-mail: xyliu@ime.pku.edu.cn [Institute of Microelectronics, Peking University, Beijing, 100871 (China)

    2010-12-15

    The accumulation process of trapped charges in a TANOS cell during P/E cycling is investigated via numerical simulation. The recombination process between trapped charges is an important issue on the retention of charge trapping memory. Our results show that accumulated trapped holes during P/E cycling can have an influence on retention, and the recombination mechanism between trapped charges should be taken into account when evaluating the retention capability of TANOS. (semiconductor devices)

  10. Efficacy of commercial traps and food odor attractants for mass trapping of Anastrepha ludens (Diptera: Tephritidae).

    Science.gov (United States)

    Lasa, Rodrigo; Velázquez, Olinda E; Ortega, Rafael; Acosta, Emilio

    2014-02-01

    One of the most important factors for the success of a mass trapping strategy to control a fruit fly involves the selection of an effective trap-lure combination. Because different species of fruit flies respond differently to the physical characteristics of a trap and to bait volatiles, the evaluation of commercial traps and lures that have proved useful against other tephtrids is necessary to determine their efficacy for mass trapping of Anastrepha ludens (Loew) (Diptera: Tephritidae). Under caged conditions, a commercial hemispherical trap with lateral holes (Maxitrap Plus) proved more attractive to A. ludens (both sexes) than five other commercial traps that were all baited with hydrolyzed protein. Among these traps, bottom invaginated traps and traps with invaginated lateral holes constructed with transparent cylinders had the best physical retention properties. When evaluated under field conditions, the lure was critical for the efficacy of the trap, and one of the traps that performed poorly in attraction and retention cage tests (MS2) resulted as one of the most effective traps when baited with CeraTrap lure. Considering the use of different trap models under field conditions, CeraTrap liquid bait was more effective in A. ludens capture than Biolure dry synthetic bait, but both lures were not replaced during the entire course of the experiment. The percentage of captured females was also slightly higher using CeraTrap lure (67.2%) than using Biolure baits (54.5-58.8%). In field tests, 75-81% of females were mated and no significant differences were observed among trap-lure combinations. Trap selectivity against nontarget adult lacewings also differed among trap-lure combinations.

  11. SEMICONDUCTOR DEVICES Effect of trapped charge accumulation on the retention of charge trapping memory

    Science.gov (United States)

    Rui, Jin; Xiaoyan, Liu; Gang, Du; Jinfeng, Kang; Ruqi, Han

    2010-12-01

    The accumulation process of trapped charges in a TANOS cell during P/E cycling is investigated via numerical simulation. The recombination process between trapped charges is an important issue on the retention of charge trapping memory. Our results show that accumulated trapped holes during P/E cycling can have an influence on retention, and the recombination mechanism between trapped charges should be taken into account when evaluating the retention capability of TANOS.

  12. Evaluating harvest-based control of invasive fish with telemetry: Performance of sea lamprey traps in the Great Lakes

    Science.gov (United States)

    Holbrook, Christopher; Bergstedt, Roger A.; Barber, Jessica M.; Bravener, Gale A; Jones, Michael L.; Krueger, Charles C.

    2016-01-01

    Physical removal (e.g., harvest via traps or nets) of mature individuals may be a cost-effective or socially acceptable alternative to chemical control strategies for invasive species, but requires knowledge of the spatial distribution of a population over time. We used acoustic telemetry to determine the current and possible future role of traps to control and assess invasive sea lampreys, Petromyzon marinus, in the St. Marys River, the connecting channel between Lake Superior and Lake Huron. Exploitation rates (i.e., fractions of an adult sea lamprey population removed by traps) at two upstream locations were compared among three years and two points of entry to the system. Telemetry receivers throughout the drainage allowed trap performance (exploitation rate) to be partitioned into two components: proportion of migrating sea lampreys that visited trap sites (availability) and proportion of available sea lampreys that were caught by traps (local trap efficiency). Estimated exploitation rates were well below those needed to provide population control in the absence of lampricides and were limited by availability and local trap efficiency. Local trap efficiency estimates for acoustic-tagged sea lampreys were lower than analogous estimates regularly obtained using traditional mark–recapture methods, suggesting that abundance had been previously underestimated. Results suggested major changes would be required to substantially increase catch, including improvements to existing traps, installation of new traps, or other modifications to attract and retain more sea lampreys. This case study also shows how bias associated with telemetry tags can be estimated and incorporated in models to improve inferences about parameters that are directly relevant to fishery management.

  13. Potential benefits, limitations and target product-profiles of odor-baited mosquito traps for malaria control in Africa.

    Directory of Open Access Journals (Sweden)

    Fredros O Okumu

    Full Text Available BACKGROUND: Traps baited with synthetic human odors have been proposed as suitable technologies for controlling malaria and other mosquito-borne diseases. We investigated the potential benefits of such traps for preventing malaria transmission in Africa and the essential characteristics that they should possess so as to be effective. METHODS AND PRINCIPAL FINDINGS: An existing mathematical model was reformulated to distinguish availability of hosts for attack by mosquitoes from availability of blood per se. This adaptation allowed the effects of pseudo-hosts such as odor-baited mosquito traps, which do not yield blood but which can nonetheless be attacked by the mosquitoes, to be simulated considering communities consisting of users and non-users of insecticide-treated nets (ITNs, currently the primary malaria prevention method. We determined that malaria transmission declines as trap coverage (proportion of total availability of all hosts and pseudo hosts that traps constitute increases. If the traps are more attractive than humans and are located in areas where mosquitoes are most abundant, 20-130 traps per 1000 people would be sufficient to match the impact of 50% community-wide ITN coverage. If such traps are used to complement ITNs, malaria transmission can be reduced by 99% or more in most scenarios representative of Africa. However, to match cost-effectiveness of ITNs, the traps delivery, operation and maintenance would have to cost a maximum of US$4.25 to 27.61 per unit per year. CONCLUSIONS AND SIGNIFICANCE: Odor-baited mosquito traps might potentially be effective and affordable tools for malaria control in Africa, particularly if they are used to complement, rather than replace, existing methods. We recommend that developers should focus on super-attractive baits and cheaper traps to enhance cost-effectiveness, and that the most appropriate way to deploy such technologies is through vertical delivery mechanisms.

  14. Towards an early warning system for Rhodesian sleeping sickness in savannah areas: man-like traps for tsetse flies.

    Directory of Open Access Journals (Sweden)

    Glyn A Vale

    Full Text Available BACKGROUND: In the savannahs of East and Southern Africa, tsetse flies (Glossina spp. transmit Trypanosoma brucei rhodesiense which causes Rhodesian sleeping sickness, the zoonotic form of human African trypanosomiasis. The flies feed mainly on wild and domestic animals and are usually repelled by humans. However, this innate aversion to humans can be undermined by environmental stresses on tsetse populations, so increasing disease risk. To monitor changes in risk, we need traps designed specifically to quantify the responsiveness of savannah tsetse to humans, but the traps currently available are designed to simulate other hosts. METHODOLOGY/PRINCIPAL FINDINGS: In Zimbabwe, two approaches were made towards developing a man-like trap for savannah tsetse: either modifying an ox-like trap or creating new designs. Tsetse catches from a standard ox-like trap used with and without artificial ox odor were reduced by two men standing nearby, by an average of 34% for Glossina morsitans morsitans and 56% for G. pallidipes, thus giving catches more like those made by hand-nets from men. Sampling by electrocuting devices suggested that the men stopped flies arriving near the trap and discouraged trap-entering responses. Most of human repellence was olfactory, as evidenced by the reduction in catches when the trap was used with the odor of hidden men. Geranyl acetone, known to occur in human odor, and dispensed at 0.2 mg/h, was about as repellent as human odor but not as powerfully repellent as wood smoke. New traps looking and smelling like men gave catches like those from men. CONCLUSION/SIGNIFICANCE: Catches from the completely new man-like traps seem too small to give reliable indices of human repellence. Better indications would be provided by comparing the catches of an ox-like trap either with or without artificial human odor. The chemistry and practical applications of the repellence of human odor and smoke deserve further study.

  15. Algae commensal community in Genlisea traps

    Directory of Open Access Journals (Sweden)

    Konrad Wołowski

    2011-01-01

    Full Text Available The community of algae occurring in Genlisea traps and on the external traps surface in laboratory conditions were studied. A total of 29 taxa were found inside the traps, with abundant diatoms, green algae (Chlamydophyceae and four morphotypes of chrysophytes stomatocysts. One morphotype is described as new for science. There are two ways of algae getting into Genlisea traps. The majority of those recorded inside the traps, are mobile; swimming freely by flagella or moving exuding mucilage like diatoms being ablate to colonize the traps themselves. Another possibility is transport of algae by invertebrates such as mites and crustaceans. In any case algae in the Genlisea traps come from the surrounding environment. Two dominant groups of algae (Chladymonas div. and diatoms in the trap environment, show ability to hydrolyze phosphomonoseters. We suggest that algae in carnivorous plant traps can compete with plant (host for organic phosphate (phosphomonoseters. From the spectrum and ecological requirements of algal species found in the traps, environment inside the traps seems to be acidic. However, further studies are needed to test the relations between algae and carnivorous plants both in laboratory conditions and in the natural environment. All the reported taxa are described briefly and documented with 74 LM and SEM micrographs.

  16. Multipole Traps as Tools in Environmental Studies

    CERN Document Server

    Mihalcea, Bogdan M; Giurgiu, Liviu C; Groza, Andreea; Surmeian, Agavni; Ganciu, Mihai; Filinov, Vladimir; Lapitsky, Dmitry; Deputatova, Lidiya; Vasilyak, Leonid; Pecherkin, Vladimir; Vladimirov, Vladimir; Syrovatka, Roman

    2015-01-01

    Trapping of microparticles, nanoparticles and aerosols is an issue of major interest for physics and chemistry. We present a setup intended for microparticle trapping in multipole linear Paul trap geometries, operating under Standard Ambient Temperature and Pressure (SATP) conditions. A 16-electrode linear trap geometry has been designed and tested, with an aim to confine a larger number of particles with respect to quadrupole traps and thus enhance the signal to noise ratio, as well as to study microparticle dynamical stability in electrodynamic fields. Experimental tests and numerical simulations suggest that multipole traps are very suited for high precision mass spectrometry measurements in case of different microparticle species or to identify the presence of certain aerosols and polluting agents in the atmosphere. Particle traps represent versatile tools for environment monitoring or for the study of many-body Coulomb systems and dusty plasmas.

  17. The Aarhus Ion Micro-Trap Project

    DEFF Research Database (Denmark)

    Miroshnychenko, Yevhen; Nielsen, Otto; Poulsen, Gregers

    As part of our involvement in the EU MICROTRAP project, we have designed, manufactured and assembled a micro-scale ion trap with integrated optical fibers. These prealigned fibers will allow delivering cooling laser light to single ions. Therefore, such a trap will not require any direct optical...... access for laser cooling. All the parts for the trap have been made in our institute [1]. The electrodes and the spacers were laser cut in the collaboration with the group of P.  Balling. In our group we have developed a technique to manufacture lensed optical fibers. The trap is now assembled...... and installed in an ultra high vacuum chamber, which includes an ablation oven for all-optical loading of the trap [2]. The next steps on the project are to demonstrate the operation of the micro-trap and the cooling of ions using fiber delivered light. [1] D. Grant, Development of Micro-Scale Ion traps, Master...

  18. Field Responses of Anopheles gambiae Complex (Diptera: Culicidae) in Liberia using Yeast-Generated Carbon Dioxide and Synthetic Lure-Baited Light Traps

    Science.gov (United States)

    2013-07-01

    VECTOR-BORNE DISEASES, SURVEILLANCE, PREVENTION Field Responses of Anopheles gambiae Complex (Diptera: Culicidae) in Liberia using Yeast-Generated...and light, bed-net, tent, andodor-baited traps (Mboera 2005). TheCenters forDiseaseControl and Prevention (CDC) light trap with its typical 4Ð6 W...using paper- clips. Although primarily developed and used to attract day ßying Stegomyia ( Aedes ) mosquitoes, blends of this lureÕs primary ingredients

  19. Modeling and Optimizing RF Multipole Ion Traps

    Science.gov (United States)

    Fanghaenel, Sven; Asvany, Oskar; Schlemmer, Stephan

    2016-06-01

    Radio frequency (rf) ion traps are very well suited for spectroscopy experiments thanks to the long time storage of the species of interest in a well defined volume. The electrical potential of the ion trap is determined by the geometry of its electrodes and the applied voltages. In order to understand the behavior of trapped ions in realistic multipole traps it is necessary to characterize these trapping potentials. Commercial programs like SIMION or COMSOL, employing the finite difference and/or finite element method, are often used to model the electrical fields of the trap in order to design traps for various purposes, e.g. introducing light from a laser into the trap volume. For a controlled trapping of ions, e.g. for low temperature trapping, the time dependent electrical fields need to be known to high accuracy especially at the minimum of the effective (mechanical) potential. The commercial programs are not optimized for these applications and suffer from a number of limitations. Therefore, in our approach the boundary element method (BEM) has been employed in home-built programs to generate numerical solutions of real trap geometries, e.g. from CAD drawings. In addition the resulting fields are described by appropriate multipole expansions. As a consequence, the quality of a trap can be characterized by a small set of multipole parameters which are used to optimize the trap design. In this presentation a few example calculations will be discussed. In particular the accuracy of the method and the benefits of describing the trapping potentials via multipole expansions will be illustrated. As one important application heating effects of cold ions arising from non-ideal multipole fields can now be understood as a consequence of imperfect field configurations.

  20. Price smarter on the Net.

    Science.gov (United States)

    Baker, W; Marn, M; Zawada, C

    2001-02-01

    Companies generally have set prices on the Internet in two ways. Many start-ups have offered untenably low prices in a rush to capture first-mover advantage. Many incumbents have simply charged the same prices on-line as they do off-line. Either way, companies are missing a big opportunity. The fundamental value of the Internet lies not in lowering prices or making them consistent but in optimizing them. After all, if it's easy for customers to compare prices on the Internet, it's also easy for companies to track customers' behavior and adjust prices accordingly. The Net lets companies optimize prices in three ways. First, it lets them set and announce prices with greater precision. Different prices can be tested easily, and customers' responses can be collected instantly. Companies can set the most profitable prices, and they can tap into previously hidden customer demand. Second, because it's so easy to change prices on the Internet, companies can adjust prices in response to even small fluctuations in market conditions, customer demand, or competitors' behavior. Third, companies can use the clickstream data and purchase histories that it collects through the Internet to segment customers quickly. Then it can offer segment-specific prices or promotions immediately. By taking full advantage of the unique possibilities afforded by the Internet to set prices with precision, adapt to changing circumstances quickly, and segment customers accurately, companies can get their pricing right. It's one of the ultimate drivers of e-business success.

  1. Petri nets and other models of concurrency

    DEFF Research Database (Denmark)

    Nielsen, Mogens; Sassone, Vladimiro

    1998-01-01

    This paper retraces, collects, and summarises contributions of the authors - in collaboration with others - on the theme of Petri nets and their categorical relationships to other models of concurrency.......This paper retraces, collects, and summarises contributions of the authors - in collaboration with others - on the theme of Petri nets and their categorical relationships to other models of concurrency....

  2. Incremental Net Effects in Multiple Regression

    Science.gov (United States)

    Lipovetsky, Stan; Conklin, Michael

    2005-01-01

    A regular problem in regression analysis is estimating the comparative importance of the predictors in the model. This work considers the 'net effects', or shares of the predictors in the coefficient of the multiple determination, which is a widely used characteristic of the quality of a regression model. Estimation of the net effects can be a…

  3. A Brief Introduction to Coloured Petri Nets

    DEFF Research Database (Denmark)

    Jensen, Kurt

    1997-01-01

    Coloured Petri Nets (CP-nets or CPN) is a graphical oriented language for design, specification, simulation and verification of systems. It is in particular well- suited for systems in which communication, synchronisation and resource sharing are important. Typical examples of application areas a...

  4. Net Tuition Trends in the United States

    Science.gov (United States)

    Gillen, Andrew; Coleman, James; Zhong, Hans

    2008-01-01

    This report documents the latest trends in net tuition for American higher education. Affordability has become a topic of concern for many, but there is a lack of information on the relevant concepts of tuition, primarily published vs. net. This report seeks to shed light on this topic. While few doubt that published tuition rates (sticker price)…

  5. Reachability via Compositionality in Petri nets

    OpenAIRE

    Sobocinski, Paweł; Stephens, Owen

    2013-01-01

    We introduce a novel technique for checking reachability in Petri nets that relies on a recently introduced compositional algebra of nets. We prove that the technique is correct, and discuss our implementation. We report promising experimental results on some well-known examples.

  6. Net analyte signal based statistical quality control

    NARCIS (Netherlands)

    Skibsted, E.T.S.; Boelens, H.F.M.; Westerhuis, J.A.; Smilde, A.K.; Broad, N.W.; Rees, D.R.; Witte, D.T.

    2005-01-01

    Net analyte signal statistical quality control (NAS-SQC) is a new methodology to perform multivariate product quality monitoring based on the net analyte signal approach. The main advantage of NAS-SQC is that the systematic variation in the product due to the analyte (or property) of interest is sep

  7. Link-Net : improving railcar fleet management

    Energy Technology Data Exchange (ETDEWEB)

    Mancini, G.; Partington, M. [Procor Limited, Oakville, ON (Canada)

    1997-12-31

    Information about Link-Net, a new rail car information facility available on Netscape, was presented. Link-Net was developed by Ontario`s Procor Limited. It can be used to find a comprehensive list of data on rail fleet management, down to details about the status of single cars. 6 figs.

  8. Net Shape Manufacturing of Aeroengine Components

    Science.gov (United States)

    2006-05-01

    Direct laser fabrication (DLF) and Net shape HIPping are two net shape manufacturing technologies which are being studied in the IRC. The direct... laser fabrication technique can produce 3-D components from their CAD files from metal powder and using a laser, the movement of which follows the paths

  9. 78 FR 72393 - Net Investment Income Tax

    Science.gov (United States)

    2013-12-02

    ... Investment Income Tax; Final and Proposed Rules #0;#0;Federal Register / Vol. 78, No. 231 / Monday, December... Parts 1 and 602 RIN 1545-BK44 Net Investment Income Tax AGENCY: Internal Revenue Service (IRS), Treasury... Investment Income Tax and the computation of Net Investment Income. The regulations affect...

  10. Using the MVC architecture on . NET platform

    OpenAIRE

    Ježek, David

    2011-01-01

    This thesis deals with usage of MVC (Model View Controller) technology in web development on ASP.NET platform from Microsoft. Mainly it deals with latest version of framework ASP.NET MVC 3. First part describes MVC architecture and the second describes usage of MVC in certain parts of web application an comparing with PHP.

  11. Policy for the Net and the Internet.

    Science.gov (United States)

    Braman, Sandra

    1995-01-01

    Reviews the literature on Internet information policy: an awareness of new policy issues, attempts to apply existing legal regulations to new processes, and the need for new policies for the Net. Explores policy issues that affect the structure of government, the economy, society, designers of the Net, and international issues. Contains 658…

  12. Asynchronous stream processing with S-Net

    NARCIS (Netherlands)

    Grelck, C.; Scholz, S.-B.; Shafarenko, A.

    2010-01-01

    We present the rationale and design of S-Net, a coordination language for asynchronous stream processing. The language achieves a near-complete separation between the application code, written in any conventional programming language, and the coordination/communication code written in S-Net. Our app

  13. Biotic and abiotic factors influencing growth rate and production of traps by the nematode-trapping fungus Duddingtonia flagrans when induced by Cooperia oncophora larvae

    DEFF Research Database (Denmark)

    Grønvold, J.; Wolstrup, J.; Nansen, P.

    1999-01-01

    A series of experiments on corn meal agar was carried out to evaluate the efficacy of the nematode-trapping fungus Duddingtonia flagrans in different abiotic and biotic conditions which occur in cow pats. Above a concentration of 50 parasitic larvae (L-3) cm(-2) the fungus produced a maximum...... of between 500 and 600 nets cm(-2) at 20 degrees C in 2 days on the surface of corn meal agar. There were no differences in the trap-producing capacity of three strains of D. flagrans (CIII4, CI3 and Trol A). On agar at 30 degrees and 20 degrees C, the fungus responded to Coaperia oncophora L-3 very quickly...... will be ready to attack parasitic larvae, when the oxygen tension increases as a result of, for example the activity of the coprophilic fauna. Artificial light giving 3000-3400 Lux on the surface of the agar significantly depressed the growth rate and the production of trapping nets in D, flagrans (CI3...

  14. Net Charge Fluctuation and String Fragmentation

    Institute of Scientific and Technical Information of China (English)

    SABen-Hao; CAIXu; TAIAn; ZHOUDai-Mei

    2004-01-01

    We present the simulation results of the net charge fluctuation in Au+Au collisions at √Snn=130 GeV from a dynamic model, JPCIAE, and its revisions. The simulations are done for the quark-gluon matter, the directly produced pions, the pion matter, and the hadron matter. The simulated net charge fluctuation of the quark-gluon matter is close to the thermal model prediction for the quark-gluon gas. However, the discrepancy exists comparing the simulated net charge fluctuation for directly produced pions and the pion matter with the thermal model prediction for pion gas and the resonance pion gas, respectively. The net charge fluctuation of hadron matter from default JPCIAE simulations is nearly 3.5 times larger than quark-gluon matter. A discussion is given for the net charge fluctuation as an evidence of QGP phase transition.

  15. Discrete, continuous, and hybrid petri nets

    CERN Document Server

    David, René

    2004-01-01

    Petri nets do not designate a single modeling formalism. In fact, newcomers to the field confess sometimes to be a little puzzled by the diversity of formalisms that are recognized under this "umbrella". Disregarding some extensions to the theoretical modeling capabilities, and looking at the level of abstraction of the formalisms, Condition/Event, Elementary, Place/Transition, Predicate/Transition, Colored, Object Oriented... net systems are frequently encountered in the literature. On the other side, provided with appropriate interpretative extensions, Controled Net Systems, Marking Diagrams (the Petri net generalization of State Diagrams), or the many-many variants in which time can be explicitly incorporated -Time(d), Deterministic, (Generalized) Stochastic, Fuzzy...- are defined. This represents another way to define practical formalisms that can be obtained by the "cro- product" of the two mentioned dimensions. Thus Petri nets constitute a modeling paradigm, understandable in a broad sense as "the total...

  16. Status and Outlook of CHIP-TRAP: the Central Michigan University High Precision Penning Trap

    CERN Document Server

    Redshaw, Matthew; Hawks, Paul; Gamage, Nadeesha D; Hunt, Curtis; Kandegedara, Rathnayake M E B; Ratnayake, Ishara S; Sharp, Lance

    2015-01-01

    At Central Michigan University we are developing a high-precision Penning trap mass spectrometer (CHIP-TRAP)that will focus on measurements with long-lived radioactive isotopes. CHIP-TRAP will consist of a pair of hyperbolic precision-measurement Penning traps, and a cylindrical capture/?filter trap in a 12 T magnetic field. Ions will be produced by external ion sources, including a laser ablation source, and transported to the capture trap at low energies enabling ions of a given m=q ratio to be selected via their time-of-flight. In the capture trap, contaminant ions will be removed with a mass-selective rf dipole excitation and the ion of interest will be transported to the measurement traps. A phase-sensitive image charge detection technique will be used for simultaneous cyclotron frequency measurements on single ions in the two precision traps, resulting in a reduction in statistical uncertainty due to magnetic field fluctuations.

  17. Discriminating between antihydrogen and mirror-trapped antiprotons in a minimum-B trap

    CERN Document Server

    Amole, C; Ashkezari, M D; Baquero-Ruiz, M; Bertsche, W; Butler, E; Cesar, C L; Chapman, S; Charlton, M; Deller, A; Eriksson, S; Fajans, J; Friesen, T; Fujiwara, M C; Gill, D R; Gutierrez, A; Hangst, J S; Hardy, W N; Hayden, M E; Humphries, A J; Hydomako, R; Kurchaninov, L; Jonsell, S; Madsen, N; Menary, S; Nolan, P; Olchanski, K; Olin, A; Povilus, A; Pusa, P; Robicheaux, F; Sarid, E; Silveira, D M; So, C; Storey, J W; Thompson, R I; van der Werf, D P; Wurtele, J S

    2012-01-01

    Recently, antihydrogen atoms were trapped at CERN in a magnetic minimum (minimum-B) trap formed by superconducting octupole and mirror magnet coils. The trapped antiatoms were detected by rapidly turning off these magnets, thereby eliminating the magnetic minimum and releasing any antiatoms contained in the trap. Once released, these antiatoms quickly hit the trap wall, whereupon the positrons and antiprotons in the antiatoms annihilated. The antiproton annihilations produce easily detected signals; we used these signals to prove that we trapped antihydrogen. However, our technique could be confounded by mirror-trapped antiprotons, which would produce seemingly-identical annihilation signals upon hitting the trap wall. In this paper, we discuss possible sources of mirror-trapped antiprotons and show that antihydrogen and antiprotons can be readily distinguished, often with the aid of applied electric fields, by analyzing the annihilation locations and times. We further discuss the general properties of antipr...

  18. Neural Net Safety Monitor Design

    Science.gov (United States)

    Larson, Richard R.

    2007-01-01

    The National Aeronautics and Space Administration (NASA) at the Dryden Flight Research Center (DFRC) has been conducting flight-test research using an F-15 aircraft (figure 1). This aircraft has been specially modified to interface a neural net (NN) controller as part of a single-string Airborne Research Test System (ARTS) computer with the existing quad-redundant flight control system (FCC) shown in figure 2. The NN commands are passed to FCC channels 2 and 4 and are cross channel data linked (CCDL) to the other computers as shown. Numerous types of fault-detection monitors exist in the FCC when the NN mode is engaged; these monitors would cause an automatic disengagement of the NN in the event of a triggering fault. Unfortunately, these monitors still may not prevent a possible NN hard-over command from coming through to the control laws. Therefore, an additional and unique safety monitor was designed for a single-string source that allows authority at maximum actuator rates but protects the pilot and structural loads against excessive g-limits in the case of a NN hard-over command input. This additional monitor resides in the FCCs and is executed before the control laws are computed. This presentation describes a floating limiter (FL) concept1 that was developed and successfully test-flown for this program (figure 3). The FL computes the rate of change of the NN commands that are input to the FCC from the ARTS. A window is created with upper and lower boundaries, which is constantly floating and trying to stay centered as the NN command rates are changing. The limiter works by only allowing the window to move at a much slower rate than those of the NN commands. Anywhere within the window, however, full rates are allowed. If a rate persists in one direction, it will eventually hit the boundary and be rate-limited to the floating limiter rate. When this happens, a persistent counter begins and after a limit is reached, a NN disengage command is generated. The

  19. Pro visual C++/CLI and the net 35 platform

    CERN Document Server

    Fraser, Stephen

    2008-01-01

    Pro Visual C++/CLI and the .NET 3.5 Platform is about writing .NET applications using C++/CLI. While readers are learning the ins and outs of .NET application development, they will also be learning the syntax of C++, both old and new to .NET. Readers will also gain a good understanding of the .NET architecture. This is truly a .NET book applying C++ as its development language not another C++ syntax book that happens to cover .NET.

  20. Vertebrate extracellular preovulatory and postovulatory egg coats.

    Science.gov (United States)

    Menkhorst, Ellen; Selwood, Lynne

    2008-11-01

    Extracellular egg coats deposited by maternal or embryonic tissues surround all vertebrate conceptuses during early development. In oviparous species, the time of hatching from extracellular coats can be considered equivalent to the time of birth in viviparous species. Extracellular coats must be lost during gestation for implantation and placentation to occur in some viviparous species. In the most recent classification of vertebrate extracellular coats, Boyd and Hamilton (Cleavage, early development and implantation of the egg. In: Parkes AS (ed.), Marshall's Physiology of Reproduction, vol. 2, 3rd ed. London: Longmans, Green & Co; 1961:1-126) defined the coat synthesized by the oocyte during oogenesis as primary and the coat deposited by follicle cells surrounding the oocyte as secondary. Tertiary egg coats are those synthesized and deposited around the primary or secondary coat by the maternal reproductive tract. This classification is difficult to reconcile with recent data collected using modern molecular biological techniques that can accurately establish the site of coat precursor synthesis and secretion. We propose that a modification to the classification by Boyd and Hamilton is required. Vertebrate egg coats should be classed as belonging to the following two broad groups: the preovulatory coat, which is deposited during oogenesis by the oocyte or follicle cells, and the postovulatory coats, which are deposited after fertilization by the reproductive tract or conceptus. This review discusses the origin and classification of vertebrate extracellular preovulatory and postovulatory coats and illustrates what is known about coat homology between the vertebrate groups.