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Sample records for acid sphingomyelinase asm

  1. Crystal structure of mammalian acid sphingomyelinase.

    PubMed

    Gorelik, Alexei; Illes, Katalin; Heinz, Leonhard X; Superti-Furga, Giulio; Nagar, Bhushan

    2016-01-01

    Acid sphingomyelinase (ASMase, ASM, SMPD1) converts sphingomyelin into ceramide, modulating membrane properties and signal transduction. Inactivating mutations in ASMase cause Niemann-Pick disease, and its inhibition is also beneficial in models of depression and cancer. To gain a better understanding of this critical therapeutic target, we determined crystal structures of mammalian ASMase in various conformations. The catalytic domain adopts a calcineurin-like fold with two zinc ions and a hydrophobic track leading to the active site. Strikingly, the membrane interacting saposin domain assumes either a closed globular conformation independent from the catalytic domain, or an open conformation, which establishes an interface with the catalytic domain essential for activity. Structural mapping of Niemann-Pick mutations reveals that most of them likely destabilize the protein's fold. This study sheds light on the molecular mechanism of ASMase function, and provides a platform for the rational development of ASMase inhibitors and therapeutic use of recombinant ASMase. PMID:27435900

  2. Crystal structure of mammalian acid sphingomyelinase

    PubMed Central

    Gorelik, Alexei; Illes, Katalin; Heinz, Leonhard X.; Superti-Furga, Giulio; Nagar, Bhushan

    2016-01-01

    Acid sphingomyelinase (ASMase, ASM, SMPD1) converts sphingomyelin into ceramide, modulating membrane properties and signal transduction. Inactivating mutations in ASMase cause Niemann–Pick disease, and its inhibition is also beneficial in models of depression and cancer. To gain a better understanding of this critical therapeutic target, we determined crystal structures of mammalian ASMase in various conformations. The catalytic domain adopts a calcineurin-like fold with two zinc ions and a hydrophobic track leading to the active site. Strikingly, the membrane interacting saposin domain assumes either a closed globular conformation independent from the catalytic domain, or an open conformation, which establishes an interface with the catalytic domain essential for activity. Structural mapping of Niemann–Pick mutations reveals that most of them likely destabilize the protein's fold. This study sheds light on the molecular mechanism of ASMase function, and provides a platform for the rational development of ASMase inhibitors and therapeutic use of recombinant ASMase. PMID:27435900

  3. Identification of novel functional inhibitors of acid sphingomyelinase.

    PubMed

    Kornhuber, Johannes; Muehlbacher, Markus; Trapp, Stefan; Pechmann, Stefanie; Friedl, Astrid; Reichel, Martin; Mühle, Christiane; Terfloth, Lothar; Groemer, Teja W; Spitzer, Gudrun M; Liedl, Klaus R; Gulbins, Erich; Tripal, Philipp

    2011-01-01

    We describe a hitherto unknown feature for 27 small drug-like molecules, namely functional inhibition of acid sphingomyelinase (ASM). These entities named FIASMAs (Functional Inhibitors of Acid SphingoMyelinAse), therefore, can be potentially used to treat diseases associated with enhanced activity of ASM, such as Alzheimer's disease, major depression, radiation- and chemotherapy-induced apoptosis and endotoxic shock syndrome. Residual activity of ASM measured in the presence of 10 µM drug concentration shows a bimodal distribution; thus the tested drugs can be classified into two groups with lower and higher inhibitory activity. All FIASMAs share distinct physicochemical properties in showing lipophilic and weakly basic properties. Hierarchical clustering of Tanimoto coefficients revealed that FIASMAs occur among drugs of various chemical scaffolds. Moreover, FIASMAs more frequently violate Lipinski's Rule-of-Five than compounds without effect on ASM. Inhibition of ASM appears to be associated with good permeability across the blood-brain barrier. In the present investigation, we developed a novel structure-property-activity relationship by using a random forest-based binary classification learner. Virtual screening revealed that only six out of 768 (0.78%) compounds of natural products functionally inhibit ASM, whereas this inhibitory activity occurs in 135 out of 2028 (6.66%) drugs licensed for medical use in humans. PMID:21909365

  4. Identification of Novel Functional Inhibitors of Acid Sphingomyelinase

    PubMed Central

    Trapp, Stefan; Pechmann, Stefanie; Friedl, Astrid; Reichel, Martin; Mühle, Christiane; Terfloth, Lothar; Groemer, Teja W.; Spitzer, Gudrun M.; Liedl, Klaus R.; Gulbins, Erich; Tripal, Philipp

    2011-01-01

    We describe a hitherto unknown feature for 27 small drug-like molecules, namely functional inhibition of acid sphingomyelinase (ASM). These entities named FIASMAs (Functional Inhibitors of Acid SphingoMyelinAse), therefore, can be potentially used to treat diseases associated with enhanced activity of ASM, such as Alzheimer's disease, major depression, radiation- and chemotherapy-induced apoptosis and endotoxic shock syndrome. Residual activity of ASM measured in the presence of 10 µM drug concentration shows a bimodal distribution; thus the tested drugs can be classified into two groups with lower and higher inhibitory activity. All FIASMAs share distinct physicochemical properties in showing lipophilic and weakly basic properties. Hierarchical clustering of Tanimoto coefficients revealed that FIASMAs occur among drugs of various chemical scaffolds. Moreover, FIASMAs more frequently violate Lipinski's Rule-of-Five than compounds without effect on ASM. Inhibition of ASM appears to be associated with good permeability across the blood-brain barrier. In the present investigation, we developed a novel structure-property-activity relationship by using a random forest-based binary classification learner. Virtual screening revealed that only six out of 768 (0.78%) compounds of natural products functionally inhibit ASM, whereas this inhibitory activity occurs in 135 out of 2028 (6.66%) drugs licensed for medical use in humans. PMID:21909365

  5. Regulation of hematogenous tumor metastasis by acid sphingomyelinase.

    PubMed

    Carpinteiro, Alexander; Becker, Katrin Anne; Japtok, Lukasz; Hessler, Gabriele; Keitsch, Simone; Požgajovà, Miroslava; Schmid, Kurt W; Adams, Constantin; Müller, Stefan; Kleuser, Burkhard; Edwards, Michael J; Grassmé, Heike; Helfrich, Iris; Gulbins, Erich

    2015-06-01

    Metastatic dissemination of cancer cells is the ultimate hallmark of malignancy and accounts for approximately 90% of human cancer deaths. We investigated the role of acid sphingomyelinase (Asm) in the hematogenous metastasis of melanoma cells. Intravenous injection of B16F10 melanoma cells into wild-type mice resulted in multiple lung metastases, while Asm-deficient mice (Smpd1(-/-) mice) were protected from pulmonary tumor spread. Transplanting wild-type platelets into Asm-deficient mice reinstated tumor metastasis. Likewise, Asm-deficient mice were protected from hematogenous MT/ret melanoma metastasis to the spleen in a mouse model of spontaneous tumor metastasis. Human and mouse melanoma cells triggered activation and release of platelet secretory Asm, in turn leading to ceramide formation, clustering, and activation of α5β1 integrins on melanoma cells finally leading to adhesion of the tumor cells. Clustering of integrins by applying purified Asm or C16 ceramide to B16F10 melanoma cells before intravenous injection restored trapping of tumor cells in the lung in Asm-deficient mice. This effect was revertable by arginine-glycine-aspartic acid peptides, which are known inhibitors of integrins, and by antibodies neutralizing β1 integrins. These findings indicate that melanoma cells employ platelet-derived Asm for adhesion and metastasis. PMID:25851537

  6. Regulation of hematogenous tumor metastasis by acid sphingomyelinase

    PubMed Central

    Carpinteiro, Alexander; Becker, Katrin Anne; Japtok, Lukasz; Hessler, Gabriele; Keitsch, Simone; Požgajovà, Miroslava; Schmid, Kurt W; Adams, Constantin; Müller, Stefan; Kleuser, Burkhard; Edwards, Michael J; Grassmé, Heike; Helfrich, Iris; Gulbins, Erich

    2015-01-01

    Metastatic dissemination of cancer cells is the ultimate hallmark of malignancy and accounts for approximately 90% of human cancer deaths. We investigated the role of acid sphingomyelinase (Asm) in the hematogenous metastasis of melanoma cells. Intravenous injection of B16F10 melanoma cells into wild-type mice resulted in multiple lung metastases, while Asm-deficient mice (Smpd1−/− mice) were protected from pulmonary tumor spread. Transplanting wild-type platelets into Asm-deficient mice reinstated tumor metastasis. Likewise, Asm-deficient mice were protected from hematogenous MT/ret melanoma metastasis to the spleen in a mouse model of spontaneous tumor metastasis. Human and mouse melanoma cells triggered activation and release of platelet secretory Asm, in turn leading to ceramide formation, clustering, and activation of α5β1 integrins on melanoma cells finally leading to adhesion of the tumor cells. Clustering of integrins by applying purified Asm or C16 ceramide to B16F10 melanoma cells before intravenous injection restored trapping of tumor cells in the lung in Asm-deficient mice. This effect was revertable by arginine-glycine-aspartic acid peptides, which are known inhibitors of integrins, and by antibodies neutralizing β1 integrins. These findings indicate that melanoma cells employ platelet-derived Asm for adhesion and metastasis. PMID:25851537

  7. Elevation of Serum Acid Sphingomyelinase Activity in Acute Kawasaki Disease.

    PubMed

    Konno, Yuuki; Takahashi, Ikuko; Narita, Ayuko; Takeda, Osamu; Koizumi, Hiromi; Tamura, Masamichi; Kikuchi, Wataru; Komatsu, Akira; Tamura, Hiroaki; Tsuchida, Satoko; Noguchi, Atsuko; Takahashi, Tsutomu

    2015-01-01

    Kawasaki disease (KD) is an acute systemic vasculitis that affects both small and medium-sized vessels including the coronary arteries in infants and children. Acid sphingomyelinase (ASM) is a lysosomal glycoprotein that hydrolyzes sphingomyelin to ceramide, a lipid, that functions as a second messenger in the regulation of cell functions. ASM activation has been implicated in numerous cellular stress responses and is associated with cellular ASM secretion, either through alternative trafficking of the ASM precursor protein or by means of an unidentified mechanism. Elevation of serum ASM activity has been described in several human diseases, suggesting that patients with diseases involving vascular endothelial cells may exhibit a preferential elevation of serum ASM activity. As acute KD is characterized by systemic vasculitis that could affect vascular endothelial cells, the elevation of serum ASM activity should be considered in these patients. In the present study, serum ASM activity in the sera of 15 patients with acute KD was determined both before and after treatment with infusion of high-dose intravenous immunoglobulin (IVIG), a first-line treatment for acute KD. Serum ASM activity before IVIG was significantly elevated in KD patients when compared to the control group (3.85 ± 1.46 nmol/0.1 ml/6 h vs. 1.15 ± 0.10 nmol/0.1 ml/6 h, p < 0.001), suggesting that ASM activation may be involved in the pathophysiology of this condition. Serum ASM activity before IVIG was significantly correlated with levels of C-reactive protein (p < 0.05). These results suggest the involvement of sphingolipid metabolism in the pathophysiology of KD. PMID:26447086

  8. Inhibition of acid sphingomyelinase by tricyclic antidepressants and analogons

    PubMed Central

    Beckmann, Nadine; Sharma, Deepa; Gulbins, Erich; Becker, Katrin Anne; Edelmann, Bärbel

    2014-01-01

    Amitriptyline, a tricyclic antidepressant, has been used in the clinic to treat a number of disorders, in particular major depression and neuropathic pain. In the 1970s the ability of tricyclic antidepressants to inhibit acid sphingomyelinase (ASM) was discovered. The enzyme ASM catalyzes the hydrolysis of sphingomyelin to ceramide. ASM and ceramide were shown to play a crucial role in a wide range of diseases, including cancer, cystic fibrosis, diabetes, Alzheimer's disease, and major depression, as well as viral (e.g., measles virus) and bacterial (e.g., Staphylococcus aureus, Pseudomonas aeruginosa) infections. Ceramide molecules may act in these diseases by the alteration of membrane biophysics, the self-association of ceramide molecules within the cell membrane and the ultimate formation of larger ceramide-enriched membrane domains/platforms. These domains were shown to serve the clustering of certain receptors such as CD95 and may also act in the above named diseases. The potential to block the generation of ceramide by inhibiting the ASM has opened up new therapeutic approaches for the treatment of these conditions. Since amitriptyline is one of the longest used clinical drugs and side effects are well studied, it could potentially become a cheap and easily accessible medication for patients suffering from these diseases. In this review, we aim to provide an overview of current in vitro and in vivo studies and clinical trials utilizing amitriptyline to inhibit ASM and contemplate possible future applications of the drug. PMID:25228885

  9. Ceramide formation mediated by acid sphingomyelinase facilitates endosomal escape of caliciviruses.

    PubMed

    Shivanna, Vinay; Kim, Yunjeong; Chang, Kyeong-Ok

    2015-09-01

    Our recent results demonstrated that bile acids facilitate virus escape from the endosomes into the cytoplasm for successful replication of porcine enteric calicivirus (PEC). We report a novel finding that bile acids can be substituted by cold treatment for endosomal escape and virus replication. This endosomal escape by cold treatment or bile acids is associated with ceramide formation by acid sphingomyelinase (ASM). ASM catalyzes hydrolysis of sphingomyelin into ceramide, which is known to destabilize lipid bilayer. Treatment of LLC-PK cells with bile acids or cold led to ceramide formation, and small molecule antagonists or siRNA of ASM blocked ceramide formation in the endosomes and significantly reduced PEC replication. Inhibition of ASM resulted in the retention of PEC, feline calicivirus or murine norovirus in the endosomes in correlation with reduced viral replication. These results suggest the importance of viral escape from the endosomes for the replication of various caliciviruses. PMID:25985440

  10. Acid Sphingomyelinase Gene Deficiency Ameliorates the Hyperhomocysteinemia-Induced Glomerular Injury in Mice

    PubMed Central

    Boini, Krishna M.; Xia, Min; Li, Caixia; Zhang, Chun; Payne, Lori P.; Abais, Justine M.; Poklis, Justin L.; Hylemon, Philip B.; Li, Pin-Lan

    2011-01-01

    Hyperhomocysteinemia (hHcys) enhances ceramide production, leading to the activation of NADPH oxidase and consequent glomerular oxidative stress and sclerosis. The present study was performed to determine whether acid sphingomyelinase (Asm), a ceramide-producing enzyme, is implicated in the development of hHcys-induced glomerular oxidative stress and injury. Uninephrectomized Asm-knockout (Asm−/−) and wild-type (Asm+/+) mice, with or without Asm short hairpin RNA (shRNA) transfection, were fed a folate-free (FF) diet for 8 weeks, which significantly elevated the plasma Hcys level compared with mice fed normal chow. By using in vivo molecular imaging, we found that transfected shRNAs were expressed in the renal cortex starting on day 3 and continued for 24 days. The FF diet significantly increased renal ceramide production, Asm mRNA and activity, urinary total protein and albumin excretion, glomerular damage index, and NADPH-dependent superoxide production in the renal cortex from Asm+/+ mice compared with that from Asm−/− or Asm shRNA-transfected wild-type mice. Immunofluorescence analysis showed that the FF diet decreased the expression of podocin but increased desmin and ceramide levels in glomeruli from Asm+/+ mice but not in those from Asm−/− and Asm shRNA-transfected wild-type mice. In conclusion, our observations reveal that Asm plays a pivotal role in mediating podocyte injury and glomerular sclerosis associated with NADPH oxidase–associated local oxidative stress during hHcys. PMID:21893018

  11. Role of acid sphingomyelinase bioactivity in human CD4+ T-cell activation and immune responses

    PubMed Central

    Bai, A; Kokkotou, E; Zheng, Y; Robson, S C

    2015-01-01

    Acid sphingomyelinase (ASM), a lipid hydrolase enzyme, has the potential to modulate various cellular activation responses via the generation of ceramide and by interaction with cellular receptors. We have hypothesized that ASM modulates CD4+ T-cell receptor activation and impacts immune responses. We first observed interactions of ASM with the intracellular domains of both CD3 and CD28. ASM further mediates T-cell proliferation after anti-CD3/CD28 antibody stimulation and alters CD4+ T-cell activation signals by generating ceramide. We noted that various pharmacological inhibitors of ASM or knockdown of ASM using small hairpin RNA inhibit CD3/CD28-mediated CD4+ T-cell proliferation and activation. Furthermore, such blockade of ASM bioactivity by biochemical inhibitors and/or molecular-targeted knockdown of ASM broadly abrogate T-helper cell responses. In conclusion, we detail immune, pivotal roles of ASM in adaptive immune T-cell responses, and propose that these pathways might provide novel targets for the therapy of autoimmune and inflammatory diseases. PMID:26203857

  12. Role of Acid Sphingomyelinase in the Regulation of Social Behavior and Memory.

    PubMed

    Zoicas, Iulia; Reichel, Martin; Gulbins, Erich; Kornhuber, Johannes

    2016-01-01

    Major depressive disorder is often associated with deficits in social and cognitive functioning. Mice transgenic for acid sphingomyelinase (t-ASM) were previously shown to have a depressive-like phenotype, which could be normalized by antidepressant treatment. Here, we investigated whether t-ASM mice show deficits in social behavior and memory performance, and whether these possible deficits might be normalized by amitriptyline treatment. Our results revealed that ASM overexpression altered the behavior of mice in a sex-dependent manner. As such, t-ASM female, but not male, mice showed an impaired social preference and a depressive- and anxiogenic-like phenotype, which could be normalized by amitriptyline treatment. Both male and female t-ASM mice showed unaltered preference for social novelty, novel object recognition, and social and object discrimination abilities. Amitriptyline treatment impaired novel object recognition and object discrimination abilities in female, but not in male, wild-type mice, while female t-ASM mice showed unaltered novel object recognition and object discrimination abilities. This study suggests that female t-ASM mice represent a model of depression with comorbid anxiety and social deficits, without memory impairments. It further suggests that ASM overexpression has a protective role against the detrimental effects of amitriptyline on female, but not on male, non-social (object) memory. PMID:27598773

  13. Thrombin stimulates albumin transcytosis in lung microvascular endothelial cells via activation of acid sphingomyelinase.

    PubMed

    Kuebler, Wolfgang M; Wittenberg, Claudia; Lee, Warren L; Reppien, Eike; Goldenberg, Neil M; Lindner, Karsten; Gao, Yizhuo; Winoto-Morbach, Supandi; Drab, Marek; Mühlfeld, Christian; Dombrowsky, Heike; Ochs, Matthias; Schütze, Stefan; Uhlig, Stefan

    2016-04-15

    Transcellular albumin transport occurs via caveolae that are abundant in lung microvascular endothelial cells. Stimulation of albumin transcytosis by proinflammatory mediators may contribute to alveolar protein leak in lung injury, yet the regulation of albumin transport and its underlying molecular mechanisms are so far incompletely understood. Here we tested the hypothesis that thrombin may stimulate transcellular albumin transport across lung microvascular endothelial cells in an acid-sphingomyelinase dependent manner. Thrombin increased the transport of fluorescently labeled albumin across confluent human lung microvascular endothelial cell (HMVEC-L) monolayers to an extent that markedly exceeds the rate of passive diffusion. Thrombin activated acid sphingomyelinase (ASM) and increased ceramide production in HMVEC-L, but not in bovine pulmonary artery cells, which showed little albumin transport in response to thrombin. Thrombin increased total caveolin-1 (cav-1) content in both whole cell lysates and lipid rafts from HMVEC-L, and this effect was blocked by inhibition of ASM or de novo protein biosynthesis. Thrombin-induced uptake of albumin into lung microvascular endothelial cells was confirmed in isolated-perfused lungs by real-time fluorescence imaging and electron microscopy of gold-labeled albumin. Inhibition of ASM attenuated thrombin-induced albumin transport both in confluent HMVEC-L and in intact lungs, whereas HMVEC-L treatment with exogenous ASM increased albumin transport and enriched lipid rafts in cav-1. Our findings indicate that thrombin stimulates transcellular albumin transport in an acid sphingomyelinase-dependent manner by inducing de novo synthesis of cav-1 and its recruitment to membrane lipid rafts. PMID:26851257

  14. Alveolar lipoproteinosis in an acid sphingomyelinase-deficient mouse model of Niemann-Pick disease.

    PubMed

    Ikegami, Machiko; Dhami, Rajwinder; Schuchman, Edward H

    2003-03-01

    Types A and B Niemann-Pick disease (NPD) are lipid storage disorders caused by the deficient activity of acid sphingomyelinase (ASM). In humans, NPD is associated with the dysfunction of numerous organs including the lung. Gene targeting of the ASM gene in transgenic mice produced an animal model with features typical of NPD, including pulmonary inflammation. To assess mechanisms by which ASM perturbed lung function, we studied lung morphology, surfactant content, and metabolism in ASM-deficient mice in vivo. Pulmonary inflammation, with increased cellular infiltrates and the accumulation of alveolar material, was associated with alterations in surfactant content. Saturated phosphatidylcholine (SatPC) content was increased twofold, and sphingomyelin content was increased 5.5-fold in lungs of the ASM knockout (ASMKO) mice. Additional sphingomyelin enhanced the sensitivity of surfactant inhibition by plasma proteins. Clearance of SatPC from the lungs of ASMKO mice was decreased. Catabolism of SatPC by alveolar macrophages from the ASMKO mouse was significantly decreased, likely accounting for decreased pulmonary SatPC in vivo. In summary, ASM is required for normal surfactant catabolism by alveolar macrophages in vivo. Alterations in surfactant composition, including increased sphingomyelin content, contributed to the abnormal surfactant function observed in the ASM-deficient mouse. PMID:12495943

  15. Acid sphingomyelinase modulates the autophagic process by controlling lysosomal biogenesis in Alzheimer’s disease

    PubMed Central

    Lee, Jong Kil; Jin, Hee Kyung; Park, Min Hee; Kim, Bo-ra; Lee, Phil Hyu; Nakauchi, Hiromitsu; Carter, Janet E.; He, Xingxuan; Schuchman, Edward H.

    2014-01-01

    In Alzheimer’s disease (AD), abnormal sphingolipid metabolism has been reported, although the pathogenic consequences of these changes have not been fully characterized. We show that acid sphingomyelinase (ASM) is increased in fibroblasts, brain, and/or plasma from patients with AD and in AD mice, leading to defective autophagic degradation due to lysosomal depletion. Partial genetic inhibition of ASM (ASM+/−) in a mouse model of familial AD (FAD; amyloid precursor protein [APP]/presenilin 1 [PS1]) ameliorated the autophagocytic defect by restoring lysosomal biogenesis, resulting in improved AD clinical and pathological findings, including reduction of amyloid-β (Aβ) deposition and improvement of memory impairment. Similar effects were noted after pharmacologic restoration of ASM to the normal range in APP/PS1 mice. Autophagic dysfunction in neurons derived from FAD patient induced pluripotent stem cells (iPSCs) was restored by partial ASM inhibition. Overall, these results reveal a novel mechanism of ASM pathogenesis in AD that leads to defective autophagy due to impaired lysosomal biogenesis and suggests that partial ASM inhibition is a potential new therapeutic intervention for the disease. PMID:25049335

  16. Acid Sphingomyelinase Gene Knockout Ameliorates Hyperhomocysteinemic Glomerular Injury in Mice Lacking Cystathionine-β-Synthase

    PubMed Central

    Boini, Krishna M.; Xia, Min; Abais, Justine M.; Xu, Ming; Li, Cai-xia; Li, Pin-Lan

    2012-01-01

    Acid sphingomyelinase (ASM) has been implicated in the development of hyperhomocysteinemia (hHcys)-induced glomerular oxidative stress and injury. However, it remains unknown whether genetically engineering of ASM gene produces beneficial or detrimental action on hHcys-induced glomerular injury. The present study generated and characterized the mice lacking cystathionine β-synthase (Cbs) and Asm mouse gene by cross breeding Cbs+/− and Asm+/− mice. Given that the homozygotes of Cbs−/−/Asm−/− mice could not survive for 3 weeks. Cbs+/−/Asm+/+, Cbs+/−/Asm+/− and Cbs+/−/Asm−/− as well as their Cbs wild type littermates were used to study the role of Asm−/− under a background of Cbs+/− with hHcys. HPLC analysis revealed that plasma Hcys level was significantly elevated in Cbs heterozygous (Cbs+/−) mice with different copies of Asm gene compared to Cbs+/+ mice with different Asm gene copies. Cbs+/−/Asm+/+ mice had significantly increased renal Asm activity, ceramide production and O2.− level compared to Cbs+/+/Asm+/+, while Cbs+/−/Asm−/− mice showed significantly reduced renal Asm activity, ceramide production and O2.− level due to increased plasma Hcys levels. Confocal microscopy demonstrated that colocalization of podocin with ceramide was much lower in Cbs+/−/Asm−/− mice compared to Cbs+/−/Asm+/+ mice, which was accompanied by a reduced glomerular damage index, albuminuria and proteinuria in Cbs+/−/Asm−/− mice. Immunofluorescent analyses of the podocin, nephrin and desmin expression also illustrated less podocyte damages in the glomeruli from Cbs+/−/Asm−/− mice compared to Cbs+/−/Asm+/+ mice. In in vitro studies of podocytes, hHcys-enhanced O2.− production, desmin expression, and ceramide production as well as decreases in VEGF level and podocin expression in podocytes were substantially attenuated by prior treatment with amitriptyline, an Asm inhibitor. In conclusion, Asm gene knockout or

  17. Acid sphingomyelinase activity is regulated by membrane lipids and facilitates cholesterol transfer by NPC2[S

    PubMed Central

    Oninla, Vincent O.; Breiden, Bernadette; Babalola, Jonathan O.; Sandhoff, Konrad

    2014-01-01

    During endocytosis, membrane components move to intraluminal vesicles of the endolysosomal compartment for digestion. At the late endosomes, cholesterol is sorted out mainly by two sterol-binding proteins, Niemann-Pick protein type C (NPC)1 and NPC2. To study the NPC2-mediated intervesicular cholesterol transfer, we developed a liposomal assay system. (Abdul-Hammed, M., B. Breiden, M. A. Adebayo, J. O. Babalola, G. Schwarzmann, and K. Sandhoff. 2010. Role of endosomal membrane lipids and NPC2 in cholesterol transfer and membrane fusion. J. Lipid Res. 51: 1747–1760.) Anionic lipids stimulate cholesterol transfer between liposomes while SM inhibits it, even in the presence of anionic bis(monoacylglycero)phosphate (BMP). Preincubation of vesicles containing SM with acid sphingomyelinase (ASM) (SM phosphodiesterase, EC 3.1.4.12) results in hydrolysis of SM to ceramide (Cer), which enhances cholesterol transfer. Besides SM, ASM also cleaves liposomal phosphatidylcholine. Anionic phospholipids derived from the plasma membrane (phosphatidylglycerol and phosphatidic acid) stimulate SM and phosphatidylcholine hydrolysis by ASM more effectively than BMP, which is generated during endocytosis. ASM-mediated hydrolysis of liposomal SM was also stimulated by incorporation of diacylglycerol (DAG), Cer, and free fatty acids into the liposomal membranes. Conversely, phosphatidylcholine hydrolysis was inhibited by incorporation of cholesterol, Cer, DAG, monoacylglycerol, and fatty acids. Our data suggest that SM degradation by ASM is required for physiological secretion of cholesterol from the late endosomal compartment, and is a key regulator of endolysosomal lipid digestion. PMID:25339683

  18. Acid sphingomyelinase regulates glucose and lipid metabolism in hepatocytes through AKT activation and AMP-activated protein kinase suppression

    PubMed Central

    Osawa, Yosuke; Seki, Ekihiro; Kodama, Yuzo; Suetsugu, Atsushi; Miura, Kouichi; Adachi, Masayuki; Ito, Hiroyasu; Shiratori, Yoshimune; Banno, Yoshiko; Olefsky, Jerrold M.; Nagaki, Masahito; Moriwaki, Hisataka; Brenner, David A.; Seishima, Mitsuru

    2011-01-01

    Acid sphingomyelinase (ASM) regulates the homeostasis of sphingolipids, including ceramides and sphingosine-1-phosphate (S1P). Because sphingolipids regulate AKT activation, we investigated the role of ASM in hepatic glucose and lipid metabolism. Initially, we overexpressed ASM in the livers of wild-type and diabetic db/db mice by adenovirus vector (Ad5ASM). In these mice, glucose tolerance was improved, and glycogen and lipid accumulation in the liver were increased. Using primary cultured hepatocytes, we confirmed that ASM increased glucose uptake, glycogen deposition, and lipid accumulation through activation of AKT and glycogen synthase kinase-3β. In addition, ASM induced up-regulation of glucose transporter 2 accompanied by suppression of AMP-activated protein kinase (AMPK) phosphorylation. Loss of sphingosine kinase-1 (SphK1) diminished ASM-mediated AKT phosphorylation, but exogenous S1P induced AKT activation in hepatocytes. In contrast, SphK1 deficiency did not affect AMPK activation. These results suggest that the SphK/S1P pathway is required for ASM-mediated AKT activation but not for AMPK inactivation. Finally, we found that treatment with high-dose glucose increased glycogen deposition and lipid accumulation in wild-type hepatocytes but not in ASM−/− cells. This result is consistent with glucose intolerance in ASM−/− mice. In conclusion, ASM modulates AKT activation and AMPK inactivation, thus regulating glucose and lipid metabolism in the liver.—Osawa, Y., Seki, E., Kodama, Y., Suetsugu, A., Miura, K., Adachi, M., Ito, H., Shiratori, Y., Banno, Y., Olefsky, J. M., Nagaki, M., Moriwaki, H., Brenner, D. A., Seishima, M. Acid sphingomyelinase regulates glucose and lipid metabolism in hepatocytes through AKT activation and AMP-activated protein kinase suppression. PMID:21163859

  19. Activation of Membrane NADPH Oxidase Associated with Lysosome-Targeted Acid Sphingomyelinase in Coronary Endothelial Cells

    PubMed Central

    Bao, Jun-Xiang; Jin, Si; Zhang, Fan; Wang, Zheng-Chao; Li, Ningjun

    2010-01-01

    Abstract This study explored the mechanism mediating the aggregation of membrane NADPH oxidase (NOX) subunits and subsequent activation of this enzyme in bovine coronary arterial endothelial cells (CAECs). With confocal microscopy, we found that FasL stimulated lipid rafts (LRs) clustering with NOX subunit aggregation and acid sphingomyelinase (ASM) gathering, which was blocked by the siRNA of sortilin, an intracellular protein responsible for the binding and targeting of ASM to lysosomes. Correspondingly, FasL-induced O2·− production through NOX in LRs fractions was abolished by sortilin siRNA. Further, with flow-cytometry and fluorescence resonance energy transfer (FRET) analysis, we surprisingly demonstrated that after FasL stimulation, sortilin was exposed to cell membranes from lysosomes together with Lamp-1 and ASM, and these lysosomal components were aggregated and form a signaling complex in cell membranes. With co-immunoprecipitation, lysosomal sortilin and ASM were found to interact more strongly when CAECs were stimulated by FasL. Functionally, inhibition of either sortilin expression, lysosome function, LRs clustering, or NOX activity significantly attenuated FasL-induced decrease in nitric oxide (NO) levels. It is concluded that lysosome-targeted ASM, through sortilin, is able to traffic to and expose to cell-membrane surface, which may lead to LRs clustering and NOX activation in CAECs. Antioxid. Redox Signal. 12, 703–712. PMID:19761405

  20. Structure of Human Acid Sphingomyelinase Reveals the Role of the Saposin Domain in Activating Substrate Hydrolysis.

    PubMed

    Xiong, Zi-Jian; Huang, Jingjing; Poda, Gennady; Pomès, Régis; Privé, Gilbert G

    2016-07-31

    Acid sphingomyelinase (ASM) is a lysosomal phosphodiesterase that catalyzes the hydrolysis of sphingomyelin to produce ceramide and phosphocholine. While other lysosomal sphingolipid hydrolases require a saposin activator protein for full activity, the ASM polypeptide incorporates a built-in N-terminal saposin domain and does not require an external activator protein. Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. The saposin domain is tightly associated along an edge of the large, bowl-shaped catalytic domain and adopts an open form that exposes a hydrophobic concave surface approximately 30Å from the catalytic center. The calculated electrostatic potential of the enzyme is electropositive at the acidic pH of the lysosome, consistent with the strict requirement for the presence of acidic lipids in target membranes. Docking studies indicate that sphingomyelin binds with the ceramide-phosphate group positioned at the binuclear zinc center and molecular dynamic simulations indicate that the intrinsic flexibility of the saposin domain is important for monomer-dimer exchange and for membrane interactions. Overall, ASM uses a combination of electrostatic and hydrophobic interactions to cause local disruptions of target bilayers in order to bring the lipid headgroup to the catalytic center in a membrane-bound reaction. PMID:27349982

  1. Instigation of NLRP3 inflammasome activation and glomerular injury in mice on the high fat diet: role of acid sphingomyelinase gene.

    PubMed

    Boini, Krishna M; Xia, Min; Koka, Saisudha; Gehr, Todd W; Li, Pin-Lan

    2016-04-01

    Ceramide has been reported to initiate inflammasome formation and activation in obesity and different pathological conditions. The present study was performed to explore the role of acid sphingomyelinase (Asm) in the development of high fat diet (HFD)-induced inflammasome and activation and consequent glomerular injury. Asm knockout (Asm(-/-)) and wild type (Asm(+/+)) mice with or without Asm short hairpin RNA (shRNA) transfection were fed a HFD or normal chow for 12 weeks to produce obesity and associated glomerular injury. HFD significantly enhanced the Asm activity, ceramide production, colocalization of Nlrp3 (Nod-like receptor protein 3) with ASC (apoptosis-associated speck-like protein) or Caspase-1, NADPH-dependent superoxide (O2(•-)) production in glomeruli of Asm(+/+) mice than in control diet-fed mice. However, such HFD-induced increases in Asm activity, ceramide production, colocalization of Nlrp3 with ASC or Caspase-1, superoxide (O(2•-)) production was attenuated in Asm(-/-) or Asm shRNA-transfected wild-type mice. In consistency with decreased inflammasome formation, the caspase-1 activity and IL-1β production was significantly attenuated in Asm(-/-) or Asm shRNA-transfected wild-type mice fed a HFD. Morphological examinations showed that HFD-induced profound injury in glomeruli of Asm(+/+) mice which was markedly attenuated in Asm(-/-) mice. The decreased glomerular damage index in Asm(-/-) mice was accompanied by attenuated proteinuria. Fluorescent immunohistochemical examinations using podocin as a podocyte marker showed that inflammasome formation induced by the HFD were mostly located in podocytes as demonstrated by co-localization of podocin with Nlrp3. In conclusion, these observations disclose a pivotal role of Asm in the HFD-induced inflammasome formation and consequent glomerular inflammation and injury. PMID:26980705

  2. Alterations of the Ceramide Metabolism in the Peri-Infarct Cortex Are Independent of the Sphingomyelinase Pathway and Not Influenced by the Acid Sphingomyelinase Inhibitor Fluoxetine.

    PubMed

    Brunkhorst, R; Friedlaender, F; Ferreirós, N; Schwalm, S; Koch, A; Grammatikos, G; Toennes, S; Foerch, C; Pfeilschifter, J; Pfeilschifter, W

    2015-01-01

    Ceramides induce important intracellular signaling pathways, modulating proliferation, migration, apoptosis, and inflammation. However, the relevance of the ceramide metabolism in the reconvalescence phase after stroke is unclear. Besides its well-known property as a selective serotonin reuptake inhibitor, fluoxetine has been reported to inhibit the acid sphingomyelinase (ASM), a key regulator of ceramide levels which derives ceramide from sphingomyelin. Furthermore, fluoxetine has shown therapeutic potential in a randomized controlled rehabilitation trial in stroke patients. Our aim was to investigate and modulate ceramide concentrations in the peri-infarct cortex, whose morphological and functional properties correlate with long-term functional outcome in stroke. We show that certain ceramide species are modulated after experimental stroke and that these changes do not result from alterations of ASM activity, but rather from nontranscriptional induction of the ceramide de novo pathway. Unexpectedly, although reducing lesion size, fluoxetine did not improve functional outcome in our model and had no significant influence on ASM activity or the concentration of ceramides. The ceramide metabolism could emerge as a potential therapeutic target in the reconvalescence phase after stroke, as its accumulation in the peri-infarct cortex potentially influences membrane functions as well as signaling events in the tissue essential for neurological recovery. PMID:26605090

  3. Inhibition of Acid Sphingomyelinase Depletes Cellular Phosphatidylserine and Mislocalizes K-Ras from the Plasma Membrane.

    PubMed

    Cho, Kwang-Jin; van der Hoeven, Dharini; Zhou, Yong; Maekawa, Masashi; Ma, Xiaoping; Chen, Wei; Fairn, Gregory D; Hancock, John F

    2015-01-01

    K-Ras must localize to the plasma membrane for biological activity; thus, preventing plasma membrane interaction blocks K-Ras signal output. Here we show that inhibition of acid sphingomyelinase (ASM) mislocalizes both the K-Ras isoforms K-Ras4A and K-Ras4B from the plasma membrane to the endomembrane and inhibits their nanoclustering. We found that fendiline, a potent ASM inhibitor, reduces the phosphatidylserine (PtdSer) and cholesterol content of the inner plasma membrane. These lipid changes are causative because supplementation of fendiline-treated cells with exogenous PtdSer rapidly restores K-Ras4A and K-Ras4B plasma membrane binding, nanoclustering, and signal output. Conversely, supplementation with exogenous cholesterol restores K-Ras4A but not K-Ras4B nanoclustering. These experiments reveal different operational pools of PtdSer on the plasma membrane. Inhibition of ASM elevates cellular sphingomyelin and reduces cellular ceramide levels. Concordantly, delivery of recombinant ASM or exogenous ceramide to fendiline-treated cells rapidly relocalizes K-Ras4B and PtdSer to the plasma membrane. K-Ras4B mislocalization is also recapitulated in ASM-deficient Neimann-Pick type A and B fibroblasts. This study identifies sphingomyelin metabolism as an indirect regulator of K-Ras4A and K-Ras4B signaling through the control of PtdSer plasma membrane content. It also demonstrates the critical and selective importance of PtdSer to K-Ras4A and K-Ras4B plasma membrane binding and nanoscale spatial organization. PMID:26572827

  4. Inhibition of Acid Sphingomyelinase Depletes Cellular Phosphatidylserine and Mislocalizes K-Ras from the Plasma Membrane

    PubMed Central

    Cho, Kwang-jin; van der Hoeven, Dharini; Zhou, Yong; Maekawa, Masashi; Ma, Xiaoping; Chen, Wei

    2015-01-01

    K-Ras must localize to the plasma membrane for biological activity; thus, preventing plasma membrane interaction blocks K-Ras signal output. Here we show that inhibition of acid sphingomyelinase (ASM) mislocalizes both the K-Ras isoforms K-Ras4A and K-Ras4B from the plasma membrane to the endomembrane and inhibits their nanoclustering. We found that fendiline, a potent ASM inhibitor, reduces the phosphatidylserine (PtdSer) and cholesterol content of the inner plasma membrane. These lipid changes are causative because supplementation of fendiline-treated cells with exogenous PtdSer rapidly restores K-Ras4A and K-Ras4B plasma membrane binding, nanoclustering, and signal output. Conversely, supplementation with exogenous cholesterol restores K-Ras4A but not K-Ras4B nanoclustering. These experiments reveal different operational pools of PtdSer on the plasma membrane. Inhibition of ASM elevates cellular sphingomyelin and reduces cellular ceramide levels. Concordantly, delivery of recombinant ASM or exogenous ceramide to fendiline-treated cells rapidly relocalizes K-Ras4B and PtdSer to the plasma membrane. K-Ras4B mislocalization is also recapitulated in ASM-deficient Neimann-Pick type A and B fibroblasts. This study identifies sphingomyelin metabolism as an indirect regulator of K-Ras4A and K-Ras4B signaling through the control of PtdSer plasma membrane content. It also demonstrates the critical and selective importance of PtdSer to K-Ras4A and K-Ras4B plasma membrane binding and nanoscale spatial organization. PMID:26572827

  5. Green tea polyphenol epigallocatechin-O-gallate induces cell death by acid sphingomyelinase activation in chronic myeloid leukemia cells.

    PubMed

    Huang, Yuhui; Kumazoe, Motofumi; Bae, Jaehoon; Yamada, Shuhei; Takai, Mika; Hidaka, Shiori; Yamashita, Shuya; Kim, Yoonhee; Won, Yeongseon; Murata, Motoki; Tsukamoto, Shuntaro; Tachibana, Hirofumi

    2015-09-01

    An epidemiological study showed that green tea consumption is associated with a reduced risk of hematopoietic malignancy. The major green tea polyphenol epigallocatechin‑3-O-gallate (EGCG) is reported to have anticancer effects. Chronic myeloid leukemia (CML) is a major hematopoietic malignancy characterized by expansion of myeloid cells. In the present study, we showed EGCG-induced acid sphingomyelinase (ASM) activation and lipid raft clustering in CML cells. The ASM inhibitor desipramine significantly reduced EGCG-induced cell death. Protein kinase Cδ is a well‑known kinase that plays an important role in ASM activation. We observed EGCG-induced phosphorylation of protein kinase Cδ at Ser664. Importantly, EGCG-induced ASM activation was significantly reduced by pretreatment of CML cells with the soluble guanylate cyclase inhibitor NS2028, suggesting that EGCG induced ASM activation through the cyclic guanosine monophosphate (cGMP)-dependent pathway. Indeed, pharmacological inhibition of a cGMP-negative regulator enhanced the anti-CML effect of EGCG. These results indicate that EGCG-induced cell death via the cGMP/ASM pathway in CML cells. PMID:26135316

  6. Green tea polyphenol epigallocatechin-O-gallate induces cell death by acid sphingomyelinase activation in chronic myeloid leukemia cells

    PubMed Central

    HUANG, YUHUI; KUMAZOE, MOTOFUMI; BAE, JAEHOON; YAMADA, SHUHEI; TAKAI, MIKA; HIDAKA, SHIORI; YAMASHITA, SHUYA; KIM, YOONHEE; WON, YEONGSEON; MURATA, MOTOKI; TSUKAMOTO, SHUNTARO; TACHIBANA, HIROFUMI

    2015-01-01

    An epidemiological study showed that green tea consumption is associated with a reduced risk of hematopoietic malignancy. The major green tea polyphenol epigallocatechin-3-O-gallate (EGCG) is reported to have anticancer effects. Chronic myeloid leukemia (CML) is a major hematopoietic malignancy characterized by expansion of myeloid cells. In the present study, we showed EGCG-induced acid sphingomyelinase (ASM) activation and lipid raft clustering in CML cells. The ASM inhibitor desipramine significantly reduced EGCG-induced cell death. Protein kinase Cδ is a well-known kinase that plays an important role in ASM activation. We observed EGCG-induced phos-phorylation of protein kinase Cδ at Ser664. Importantly, EGCG-induced ASM activation was significantly reduced by pretreatment of CML cells with the soluble guanylate cyclase inhibitor NS2028, suggesting that EGCG induced ASM activation through the cyclic guanosine monophosphate (cGMP)-dependent pathway. Indeed, pharmacological inhibition of a cGMP-negative regulator enhanced the anti-CML effect of EGCG. These results indicate that EGCG-induced cell death via the cGMP/ASM pathway in CML cells. PMID:26135316

  7. Acid Sphingomyelinase-Derived Ceramide Regulates ICAM-1 Function during T Cell Transmigration across Brain Endothelial Cells.

    PubMed

    Lopes Pinheiro, Melissa A; Kroon, Jeffrey; Hoogenboezem, Mark; Geerts, Dirk; van Het Hof, Bert; van der Pol, Susanne M A; van Buul, Jaap D; de Vries, Helga E

    2016-01-01

    Multiple sclerosis (MS) is a chronic demyelinating disorder of the CNS characterized by immune cell infiltration across the brain vasculature into the brain, a process not yet fully understood. We previously demonstrated that the sphingolipid metabolism is altered in MS lesions. In particular, acid sphingomyelinase (ASM), a critical enzyme in the production of the bioactive lipid ceramide, is involved in the pathogenesis of MS; however, its role in the brain vasculature remains unknown. Transmigration of T lymphocytes is highly dependent on adhesion molecules in the vasculature such as intercellular adhesion molecule-1 (ICAM-1). In this article, we hypothesize that ASM controls T cell migration by regulating ICAM-1 function. To study the role of endothelial ASM in transmigration, we generated brain endothelial cells lacking ASM activity using a lentiviral shRNA approach. Interestingly, although ICAM-1 expression was increased in cells lacking ASM activity, we measured a significant decrease in T lymphocyte adhesion and consequently transmigration both in static and under flow conditions. As an underlying mechanism, we revealed that upon lack of endothelial ASM activity, the phosphorylation of ezrin was perturbed as well as the interaction between filamin and ICAM-1 upon ICAM-1 clustering. Functionally this resulted in reduced microvilli formation and impaired transendothelial migration of T cells. In conclusion, in this article, we show that ASM coordinates ICAM-1 function in brain endothelial cells by regulating its interaction with filamin and phosphorylation of ezrin. The understanding of these underlying mechanisms of T lymphocyte transmigration is of great value to develop new strategies against MS lesion formation. PMID:26597010

  8. The emerging role of acid sphingomyelinase in autophagy.

    PubMed

    Perrotta, Cristiana; Cervia, Davide; De Palma, Clara; Assi, Emma; Pellegrino, Paolo; Bassi, Maria Teresa; Clementi, Emilio

    2015-05-01

    Autophagy, the main intracellular process of cytoplasmic material degradation, is involved in cell survival and death. Autophagy is regulated at various levels and novel modulators of its function are being continuously identified. An intriguing recent observation is that among these modulators is the sphingolipid metabolising enzyme, Acid Sphingomyelinase (A-SMase), already known to play a fundamental role in apoptotic cell death participating in several pathophysiological conditions. In this review we analyse and discuss the relationship between autophagy and A-SMase describing how A-SMase may regulate it and defining, for the first time, the existence of an A-SMase-autophagy axis. The imbalance of this axis plays a role in cancer, nervous system, cardiovascular, and hepatic disorders. PMID:25666706

  9. Lack of Acid Sphingomyelinase Induces Age-Related Retinal Degeneration

    PubMed Central

    Wu, Bill X.; Fan, Jie; Boyer, Nicholas P.; Jenkins, Russell W.; Koutalos, Yiannis; Hannun, Yusuf A.; Crosson, Craig E.

    2015-01-01

    Background Mutations of acid sphingomyelinase (ASMase) cause Niemann–Pick diseases type A and B, which are fatal inherited lipid lysosomal storage diseases, characterized with visceral organ abnormalities and neurodegeneration. However, the effects of suppressing retinal ASMase expression are not understood. The goal of this study was to determine if the disruption of ASMase expression impacts the retinal structure and function in the mouse, and begin to investigate the mechanisms underlying these abnormalities. Methods Acid sphingomyelinase knockout (ASMase KO) mice were utilized to study the roles of this sphingolipid metabolizing enzyme in the retina. Electroretinogram and morphometric analysis were used to assess the retinal function and structure at various ages. Sphingolipid profile was determined by liquid chromatography-mass spectrometry. Western blots evaluated the level of the autophagy marker LC3-II. Results When compared to control animals, ASMase KO mice exhibited significant age-dependent reduction in ERG a- and b-wave amplitudes. Associated with these functional deficits, morphometric analysis revealed progressive thinning of retinal layers; however, the most prominent degeneration was observed in the photoreceptor and outer nuclear layer. Additional analyses of ASMase KO mice revealed early reduction in ERG c-wave amplitudes and increased lipofuscin accumulation in the retinal pigment epithelium (RPE). Sphingolipid analyses showed abnormal accumulation of sphingomyelin and sphingosine in ASMase KO retinas. Western blot analyses showed a higher level of the autophagosome marker LC3-II. Conclusions These studies demonstrate that ASMase is necessary for the maintenance of normal retinal structure and function. The early outer retinal dysfunction, outer segment degeneration, accumulation of lipofuscin and autophagosome markers provide evidence that disruption of lysosomal function contributes to the age-dependent retinal degeneration exhibited by

  10. Instigation of NLRP3 inflammasome activation and glomerular injury in mice on the high fat diet: role of acid sphingomyelinase gene

    PubMed Central

    Boini, Krishna M.; Xia, Min; Koka, Saisudha; Gehr, Todd W.; Li, Pin-Lan

    2016-01-01

    Ceramide has been reported to initiate inflammasome formation and activation in obesity and different pathological conditions. The present study was performed to explore the role of acid sphingomyelinase (Asm) in the development of high fat diet (HFD)-induced inflammasome and activation and consequent glomerular injury. Asm knockout (Asm−/−) and wild type (Asm+/+) mice with or without Asm short hairpin RNA (shRNA) transfection were fed a HFD or normal chow for 12 weeks to produce obesity and associated glomerular injury. HFD significantly enhanced the Asm activity, ceramide production, colocalization of Nlrp3 (Nod-like receptor protein 3) with ASC (apoptosis-associated speck-like protein) or Caspase-1, NADPH-dependent superoxide (O2•−) production in glomeruli of Asm+/+mice than in control diet-fed mice. However, such HFD-induced increases in Asm activity, ceramide production, colocalization of Nlrp3 with ASC or Caspase-1, superoxide (O2•−) production was attenuated in Asm−/− or Asm shRNA-transfected wild-type mice. In consistency with decreased inflammasome formation, the caspase-1 activity and IL-1β production was significantly attenuated in Asm−/− or Asm shRNA-transfected wild-type mice fed a HFD. Morphological examinations showed that HFD-induced profound injury in glomeruli of Asm+/+ mice which was markedly attenuated in Asm−/− mice. The decreased glomerular damage index in Asm−/− mice was accompanied by attenuated proteinuria. Fluorescent immunohistochemical examinations using podocin as a podocyte marker showed that inflammasome formation induced by the HFD were mostly located in podocytes as demonstrated by co-localization of podocin with Nlrp3. In conclusion, these observations disclose a pivotal role of Asm in the HFD-induced inflammasome formation and consequent glomerular inflammation and injury. PMID:26980705

  11. Heat Shock Protein 70.1 (Hsp70.1) Affects Neuronal Cell Fate by Regulating Lysosomal Acid Sphingomyelinase*

    PubMed Central

    Zhu, Hong; Yoshimoto, Tanihiro; Yamashima, Tetsumori

    2014-01-01

    The inducible expression of heat shock protein 70.1 (Hsp70.1) plays cytoprotective roles in its molecular chaperone function. Binding of Hsp70 to an endolysosomal phospholipid, bis(monoacylglycero)phosphate (BMP), has been recently shown to stabilize lysosomal membranes by enhancing acid sphingomyelinase (ASM) activity in cancer cells. Using the monkey experimental paradigm, we have reported that calpain-mediated cleavage of oxidized Hsp70.1 causes neurodegeneration in the hippocampal cornu ammonis 1 (CA1), whereas expression of Hsp70.1 in the motor cortex without calpain activation contributes to neuroprotection. However, the molecular mechanisms of the lysosomal destabilization/stabilization determining neuronal cell fate have not been elucidated. To elucidate whether regulation of lysosomal ASM could affect the neuronal fate, we analyzed Hsp70.1-BMP binding and ASM activity by comparing the motor cortex and the CA1. We show that Hsp70.1 being localized at the lysosomal membrane, lysosomal lipid BMP levels, and the lipid binding domain of Hsp70.1 are crucial for Hsp70.1-BMP binding. In the postischemic motor cortex, Hsp70.1 being localized at the lysosomal membrane could bind to BMP without calpain activation and decreased BMP levels, resulting in increasing ASM activity and lysosomal stability. However, in the postischemic CA1, calpain activation and a concomitant decrease in the lysosomal membrane localization of Hsp70.1 and BMP levels may diminish Hsp70.1-BMP binding, resulting in decreased ASM activity and lysosomal rupture with leakage of cathepsin B into the cytosol. A TUNEL assay revealed the differential neuronal vulnerability between the CA1 and the motor cortex. These results suggest that regulation of ASM activation in vivo by Hsp70.1-BMP affects lysosomal stability and neuronal survival or death after ischemia/reperfusion. PMID:25074941

  12. An acidic sphingomyelinase Type C activity from Mycobacterium tuberculosis.

    PubMed

    Castro-Garza, Jorge; González-Salazar, Francisco; Quinn, Frederick D; Karls, Russell K; De La Garza-Salinas, Laura Hermila; Guzmán-de la Garza, Francisco J; Vargas-Villarreal, Javier

    2016-01-01

    Sphingomyelinases (SMases) catalyze the hydrolysis of sphingomyelin to ceramide and phosphorylcholine. Sphingolipids are recognized as diverse and dynamic regulators of a multitude of cellular processes mediating cell cycle control, differentiation, stress response, cell migration, adhesion, and apoptosis. Bacterial SMases are virulence factors for several species of pathogens. Whole cell extracts of Mycobacterium tuberculosis strains H37Rv and CDC1551 were assayed using [N-methyl-(14)C]-sphingomyelin as substrate. Acidic Zn(2+)-dependent SMase activity was identified in both strains. Peak SMase activity was observed at pH 5.5. Interestingly, overall SMase activity levels from CDC1551 extracts are approximately 1/3 of those of H37Rv. The presence of exogenous SMase produced by M. tuberculosis during infection may interfere with the normal host inflammatory response thus allowing the establishment of infection and disease development. This Type C activity is different from previously identified M. tuberculosis SMases. Defining the biochemical characteristics of M. tuberculosis SMases helps to elucidate the roles that these enzymes play during infection and disease. PMID:26948102

  13. Targeting acid sphingomyelinase reduces cardiac ceramide accumulation in the post-ischemic heart.

    PubMed

    Klevstig, Martina; Ståhlman, Marcus; Lundqvist, Annika; Scharin Täng, Margareta; Fogelstrand, Per; Adiels, Martin; Andersson, Linda; Kolesnick, Richard; Jeppsson, Anders; Borén, Jan; Levin, Malin C

    2016-04-01

    Ceramide accumulation is known to accompany acute myocardial ischemia, but its role in the pathogenesis of ischemic heart disease is unclear. In this study, we aimed to determine how ceramides accumulate in the ischemic heart and to determine if cardiac function following ischemia can be improved by reducing ceramide accumulation. To investigate the association between ceramide accumulation and heart function, we analyzed myocardial left ventricle biopsies from subjects with chronic ischemia and found that ceramide levels were higher in biopsies from subjects with reduced heart function. Ceramides are produced by either de novo synthesis or hydrolysis of sphingomyelin catalyzed by acid and/or neutral sphingomyelinase. We used cultured HL-1 cardiomyocytes to investigate these pathways and showed that acid sphingomyelinase activity rather than neutral sphingomyelinase activity or de novo sphingolipid synthesis was important for hypoxia-induced ceramide accumulation. We also used mice with a partial deficiency in acid sphingomyelinase (Smpd1(+/-) mice) to investigate if limiting ceramide accumulation under ischemic conditions would have a beneficial effect on heart function and survival. Although we showed that cardiac ceramide accumulation was reduced in Smpd1(+/-) mice 24h after an induced myocardial infarction, this reduction was not accompanied by an improvement in heart function or survival. Our findings show that accumulation of cardiac ceramides in the post-ischemic heart is mediated by acid sphingomyelinase. However, targeting ceramide accumulation in the ischemic heart may not be a beneficial treatment strategy. PMID:26930027

  14. Requirement of translocated lysosomal V1 H+-ATPase for activation of membrane acid sphingomyelinase and raft clustering in coronary endothelial cells

    PubMed Central

    Xu, Ming; Xia, Min; Li, Xiao-Xue; Han, Wei-Qing; Boini, Krishna M.; Zhang, Fan; Zhang, Yang; Ritter, Joseph K; Li, Pin-Lan

    2012-01-01

    Acid sphingomyelinase (ASM) mediates the formation of membrane raft (MR) redox signalosomes in a process that depends on a local acid microenvironment in coronary arterial endothelial cells (CAECs). However, it is not known how this local acid microenvironment is formed and maintained. The present study hypothesized that lysosomal V1 H+-ATPase provides a hospitable acid microenvironment for activation of ASM when lysosomes traffic and fuse into the cell membrane. Confocal microscopy showed that local pH change significantly affected MRs, with more fluorescent patches under low pH. Correspondingly, the ASM product, ceramide, increased locally in the cell membrane. Electron spin resonance assay showed that local pH increase significantly inhibited NADPH oxidase–mediated production of O2−. in CAECs. Direct confocal microscopy demonstrated that Fas ligand resulted in localized areas of decreased pH around CAEC membranes. The inhibitors of both lysosomal fusion and H+-ATPase apparently attenuated FasL-caused pH decrease. V1 H+-ATPase accumulation and activity on cell membranes were substantially suppressed by the inhibitors of lysosomal fusion or H+-ATPase. These results provide the first direct evidence that translocated lysosomal V1 H+-ATPase critically contributes to the formation of local acid microenvironment to facilitate activation of ASM and consequent MR aggregation, forming MR redox signalosomes and mediating redox signaling in CAECs. PMID:22357614

  15. Suppression of Acid Sphingomyelinase Protects the Retina from Ischemic Injury

    PubMed Central

    Fan, Jie; Wu, Bill X.; Crosson, Craig E.

    2016-01-01

    Purpose Acid sphingomyelinase (ASMase) catalyzes the hydrolysis of sphingomyelin to ceramide and mediates multiple responses involved in inflammatory and apoptotic signaling. However, the role ASMase plays in ischemic retinal injury has not been investigated. The purpose of this study was to investigate how reduced ASMase expression impacts retinal ischemic injury. Methods Changes in ceramide levels and ASMase activity were determined by high performance liquid chromatography-tandem mass spectrometry analysis and ASMase activity. Retinal function and morphology were assessed by electroretinography (ERG) and morphometric analyses. Levels of TNF-α were determined by ELISA. Activation of p38 MAP kinase was assessed by Western blot analysis. Results In wild-type mice, ischemia produced a significant increase in retinal ASMase activity and ceramide levels. These increases were associated with functional deficits as measured by ERG analysis and significant structural degeneration in most retinal layers. In ASMase+/− mice, retinal ischemia did not significantly alter ASMase activity, and the rise in ceramide levels were significantly reduced compared to levels in retinas from wild-type mice. In ASMase+/− mice, functional and morphometric analyses of ischemic eyes revealed significantly less retinal degeneration than in injured retinas from wild-type mice. The ischemia-induced increase in retinal TNF-α levels was suppressed by the administration of the ASMase inhibitor desipramine, or by reducing ASMase expression. Conclusions Our results demonstrate that reducing ASMase expression provides partial protection from ischemic injury. Hence, the production of ceramide and subsequent mediators plays a role in the development of ischemic retinal injury. Modulating ASMase may present new opportunities for adjunctive therapies when treating retinal ischemic disorders. PMID:27571014

  16. Secretory sphingomyelinase in health and disease.

    PubMed

    Kornhuber, Johannes; Rhein, Cosima; Müller, Christian P; Mühle, Christiane

    2015-06-01

    Acid sphingomyelinase (ASM), a key enzyme in sphingolipid metabolism, hydrolyzes sphingomyelin to ceramide and phosphorylcholine. In mammals, the expression of a single gene, SMPD1, results in two forms of the enzyme that differ in several characteristics. Lysosomal ASM (L-ASM) is located within the lysosome, requires no additional Zn2+ ions for activation and is glycosylated mainly with high-mannose oligosaccharides. By contrast, the secretory ASM (S-ASM) is located extracellularly, requires Zn2+ ions for activation, has a complex glycosylation pattern and has a longer in vivo half-life. In this review, we summarize current knowledge regarding the physiology and pathophysiology of S-ASM, including its sources and distribution, molecular and cellular mechanisms of generation and regulation and relevant in vitro and in vivo studies. Polymorphisms or mutations of SMPD1 lead to decreased S-ASM activity, as detected in patients with Niemann-Pick disease B. Thus, lower serum/plasma activities of S-ASM are trait markers. No genetic causes of increased S-ASM activity have been identified. Instead, elevated activity is the result of enhanced release (e.g., induced by lipopolysaccharide and cytokine stimulation) or increased enzyme activation (e.g., induced by oxidative stress). Increased S-ASM activity in serum or plasma is a state marker of a wide range of diseases. In particular, high S-ASM activity occurs in inflammation of the endothelium and liver. Several studies have demonstrated a correlation between S-ASM activity and mortality induced by severe inflammatory diseases. Serial measurements of S-ASM reveal prolonged activation and, therefore, the measurement of this enzyme may also provide information on past inflammatory processes. Thus, S-ASM may be both a promising clinical chemistry marker and a therapeutic target. PMID:25803076

  17. A Novel Mechanism of Lysosomal Acid Sphingomyelinase Maturation

    PubMed Central

    Jenkins, Russell W.; Idkowiak-Baldys, Jolanta; Simbari, Fabio; Canals, Daniel; Roddy, Patrick; Riner, Clarke D.; Clarke, Christopher J.; Hannun, Yusuf A.

    2011-01-01

    Acid sphingomyelinase (aSMase) catalyzes the hydrolysis of sphingomyelin (SM) to form the bioactive lipid ceramide (Cer). Notably, aSMase exists in two forms: a zinc (Zn2+)-independent lysosomal aSMase (L-SMase) and a Zn2+-dependent secreted aSMase (S-SMase) that arise from alternative trafficking of a single protein precursor. Despite extensive investigation into the maturation and trafficking of aSMase, the exact identity of mature L-SMase has remained unclear. Here, we describe a novel mechanism of aSMase maturation involving C-terminal proteolytic processing within, or in close proximity to, endolysosomes. Using two different C-terminal-tagged constructs of aSMase (V5, DsRed), we demonstrate that aSMase is processed from a 75-kDa, Zn2+-activated proenzyme to a mature 65 kDa, Zn2+-independent L-SMase. L-SMase is recognized by a polyclonal Ab to aSMase, but not by anti-V5 or anti-DsRed antibodies, suggesting that the C-terminal tag is lost during maturation. Furthermore, indirect immunofluorescence staining demonstrated that mature L-SMase colocalized with the lysosomal marker LAMP1, whereas V5-aSMase localized to the Golgi secretory pathway. Moreover, V5-aSMase possessed Zn2+-dependent activity suggesting it may represent the common protein precursor of S-SMase and L-SMase. Importantly, the 65-kDa L-SMase, but not V5-aSMase, was sensitive to the lysosomotropic inhibitor desipramine, co-fractionated with lysosomes, and migrated at the same Mr as partially purified human aSMase. Finally, three aSMase mutants containing C-terminal Niemann-Pick mutations (R600H, R600P, ΔR608) exhibited defective proteolytic maturation. Taken together, these results demonstrate that mature L-SMase arises from C-terminal proteolytic processing of pro-aSMase and suggest that impaired C-terminal proteolysis may lead to severe defects in L-SMase function. PMID:21098024

  18. Improved sensitivity of an acid sphingomyelinase activity assay using a C6:0 sphingomyelin substrate.

    PubMed

    Chuang, Wei-Lien; Pacheco, Joshua; Cooper, Samantha; Kingsbury, Jonathan S; Hinds, John; Wolf, Pavlina; Oliva, Petra; Keutzer, Joan; Cox, Gerald F; Zhang, Kate

    2015-06-01

    Short-chain C6-sphingomyelin is an artificial substrate that was used in an acid sphingomyelinase activity assay for a pilot screening study of patients with Niemann-Pick disease types A and B. Using previously published multiplex and single assay conditions, normal acid sphingomyelinase activity levels (i.e. false negative results) were observed in two sisters with Niemann-Pick B who were compound heterozygotes for two missense mutations, p.C92W and p.P184L, in the SMPD1 gene. Increasing the sodium taurocholate detergent concentration in the assay buffer lowered the activity levels of these two patients into the range observed with other patients with clear separation from normal controls. PMID:26937397

  19. Improved sensitivity of an acid sphingomyelinase activity assay using a C6:0 sphingomyelin substrate

    PubMed Central

    Chuang, Wei-Lien; Pacheco, Joshua; Cooper, Samantha; Kingsbury, Jonathan S.; Hinds, John; Wolf, Pavlina; Oliva, Petra; Keutzer, Joan; Cox, Gerald F.; Zhang, Kate

    2015-01-01

    Short-chain C6-sphingomyelin is an artificial substrate that was used in an acid sphingomyelinase activity assay for a pilot screening study of patients with Niemann–Pick disease types A and B. Using previously published multiplex and single assay conditions, normal acid sphingomyelinase activity levels (i.e. false negative results) were observed in two sisters with Niemann–Pick B who were compound heterozygotes for two missense mutations, p.C92W and p.P184L, in the SMPD1 gene. Increasing the sodium taurocholate detergent concentration in the assay buffer lowered the activity levels of these two patients into the range observed with other patients with clear separation from normal controls. PMID:26937397

  20. Niemann-Pick disease: A frequent missense mutation in the acid sphingomyelinase gene of Ashkenazi Jewish type A and B patients

    SciTech Connect

    Levran, O.; Desnick, R.J.; Schuchan, E.H. )

    1991-05-01

    Although the A and B subtypes of Niemann-Pick disease (NPD) both result from the deficient activity of acid sphingomyelinase and the lysosomal accumulation of sphingomyelin, they have remarkably distinct phenotypes. Type A disease is a fatal neurodegenerative disorder of infancy, whereas type B disease has no neurologic manifestations and is characterized primarily by reticuloendothelial involvement and survival into adulthood. Both disorders are more frequent among individuals of Ashkenazi Jewish ancestry than in the general population. The recent isolation and characterization of cDNA and genomic sequences encoding ASM has facilitated investigation of the molecular lesions causing the NPD subtypes. Total RNA was reverse-transcribed, and the ASm cDNA from an Ashkenazi Jewish type A patient was specifically amplified by the polymerase chain reaction (PCR). Molecular analysis of the PCR products revealed a G{r arrow} T transversion of nucleotide 1,487, which occurred at a CpG dinucleotide and predicted an Arg{r arrow} Leu substitution and that both parents and several other relatives were heterozygous. These findings identify a frequent missense mutation among NPD patients of Ashkenazi Jewish ancestry that results in neuronopathic type A disease when homoallelic and can result in the nonneuronopathic type B phenotype when heteroallelic. The identification of this ASM mutation in Ashkenazi Jewish patients should facilitate the prevention of NPD in this population by carrier detection with molecular diagnostic techniques.

  1. Optimization of a histopathological biomarker for sphingomyelin accumulation in acid sphingomyelinase deficiency.

    PubMed

    Taksir, Tatyana V; Johnson, Jennifer; Maloney, Colleen L; Yandl, Emily; Griffiths, Denise; Thurberg, Beth L; Ryan, Susan

    2012-08-01

    Niemann-Pick disease (types A and B), or acid sphingomyelinase deficiency, is an inherited deficiency of acid sphingomyelinase, resulting in intralysosomal accumulation of sphingomyelin in cells throughout the body, particularly within those of the reticuloendothelial system. These cellular changes result in hepatosplenomegaly and pulmonary infiltrates in humans. A knockout mouse model mimics many elements of human ASMD and is useful for studying disease histopathology. However, traditional formalin-fixation and paraffin embedding of ASMD tissues dissolves sphingomyelin, resulting in tissues with a foamy cell appearance, making quantitative analysis of the substrate difficult. To optimize substrate fixation and staining, a modified osmium tetroxide and potassium dichromate postfixation method was developed to preserve sphingomyelin in epon-araldite embedded tissue and pulmonary cytology specimens. After processing, semi-thin sections were incubated with tannic acid solution followed by staining with toluidine blue/borax. This modified method provides excellent preservation and staining contrast of sphingomyelin with other cell structures. The resulting high-resolution light microscopy sections permit digital quantification of sphingomyelin in light microscopic fields. A lysenin affinity stain for sphingomyelin was also developed for use on these semi-thin epon sections. Finally, ultrathin serial sections can be cut from these same tissue blocks and stained for ultrastructural examination by electron microscopy. PMID:22614361

  2. Clearance of Hepatic Sphingomyelin by Olipudase Alfa Is Associated With Improvement in Lipid Profiles in Acid Sphingomyelinase Deficiency.

    PubMed

    Thurberg, Beth L; Wasserstein, Melissa P; Jones, Simon A; Schiano, Thomas D; Cox, Gerald F; Puga, Ana Cristina

    2016-09-01

    Acid sphingomyelinase deficiency (ASMD; Niemann-Pick disease type A and B) is a lysosomal storage disorder characterized by abnormal intracellular sphingomyelin (SM) accumulation. Prominent liver involvement results in hepatomegaly, fibrosis/cirrhosis, abnormal liver chemistries, and a proatherogenic lipid profile. Olipudase alfa (recombinant human ASM) is in clinical development as an investigational enzyme replacement therapy for the non-neurological manifestations of ASMD. In a phase 1b study conducted to evaluate the safety and tolerability of within-patient dose escalation with olipudase alfa, measurement of SM levels in liver biopsies was used as a pharmacodynamic biomarker of substrate burden. Five adult patients with non neuronopathic ASMD received escalating doses of olipudase alfa every 2 weeks for 26 weeks. Liver biopsies obtained at baseline and 26 weeks after treatment were evaluated for SM storage by histomorphometric analysis, biochemistry, and electron microscopy. Biopsies were also assessed for inflammation and fibrosis, and for the association of SM levels with liver volume, liver function tests, and lipid profiles. At baseline, SM storage present in Kupffer cells and hepatocytes ranged from 9.8% to 53.8% of the microscopic field. After 26 weeks of treatment, statistically significant reductions in SM (P<0.0001) measured by morphometry were seen in 4 patients with evaluable liver biopsies. The 26-week biopsy of the fifth patient was insufficient for morphometric quantitation. Posttreatment SM levels ranged from 1.2% to 9.5% of the microscopic field, corresponding to an 84% to 92% relative reduction from baseline. Improvements in liver volume, liver function tests, and lipid profiles were also observed. This study illustrates the utility of SM assessment by liver biopsy as a pharmacodynamic biomarker of disease burden in these patients. PMID:27340749

  3. Acid sphingomyelinase is activated in sickle cell erythrocytes and contributes to inflammatory microparticle generation in SCD.

    PubMed

    Awojoodu, Anthony O; Keegan, Philip M; Lane, Alicia R; Zhang, Yuying; Lynch, Kevin R; Platt, Manu O; Botchwey, Edward A

    2014-09-18

    Sphingolipids are a class of lipids containing a backbone of sphingoid bases that can be produced de novo through the reaction of palmitate and serine and further metabolized through the activity of various enzymes to produce intermediates with diverse roles in cellular processes and signal transduction. One of these intermediates, sphingosine 1-phosphate (S1P), is stored at high concentrations (1 μM) in red blood cells (RBCs) and directs a wide array of cellular processes mediated by 5 known G-protein coupled receptors (S1P1-S1P5). In this study, we show that RBC membrane alterations in sickle cell disease enhance the activation acid sphingomyelinase by 13%, resulting in increased production and storage of sphingosine (2.6-fold) and S1P (3.5-fold). We also show that acid sphingomyelinase enhances RBC-derived microparticle (MP) generation. These MPs are internalized by myeloid cells and promote proinflammatory cytokine secretion and endothelial cell adhesion, suggesting that potential crosstalk between circulating inflammatory cells and MPs may contribute to the inflammation-rooted pathogenesis of the disease. Treatment with amitriptyline reduces MP generation in vitro and in vivo and might be used to mitigate inflammatory processes in sickle cell disease. PMID:25075126

  4. Acidic Sphingomyelinase Controls Hepatic Stellate Cell Activation and in Vivo Liver Fibrogenesis

    PubMed Central

    Moles, Anna; Tarrats, Núria; Morales, Albert; Domínguez, Marlene; Bataller, Ramón; Caballería, Juan; García-Ruiz, Carmen; Fernández-Checa, José C.; Marí, Montserrat

    2010-01-01

    The mechanisms linking hepatocellular death, hepatic stellate cell (HSC) activation, and liver fibrosis are largely unknown. Here, we investigate whether acidic sphingomyelinase (ASMase), a known regulator of death receptor and stress-induced hepatocyte apoptosis, plays a role in liver fibrogenesis. We show that selective stimulation of ASMase (up to sixfold), but not neutral sphingomyelinase, occurs during the transdifferentiation/activation of primary mouse HSCs into myofibroblast-like cells, coinciding with cathepsin B (CtsB) and D (CtsD) processing. ASMase inhibition or genetic down-regulation by small interfering RNA blunted CtsB/D processing, preventing the activation and proliferation of mouse and human HSCs (LX2 cells). In accordance, HSCs from heterozygous ASMase mice exhibited decreased CtsB/D processing, as well as lower levels of α-smooth muscle actin expression and proliferation. Moreover, pharmacological CtsB inhibition reproduced the antagonism of ASMase in preventing the fibrogenic properties of HSCs, without affecting ASMase activity. Interestingly, liver fibrosis induced by bile duct ligation or carbon tetrachloride administration was reduced in heterozygous ASMase mice compared with that in wild-type animals, regardless of their sensitivity to liver injury in either model. To provide further evidence for the ASMase-CtsB pathway in hepatic fibrosis, liver samples from patients with nonalcoholic steatohepatitis were studied. CtsB and ASMase mRNA levels increased eight- and threefold, respectively, in patients compared with healthy controls. These findings illustrate a novel role of ASMase in HSC biology and liver fibrogenesis by regulating its downstream effectors CtsB/D. PMID:20651240

  5. Essential role for acid sphingomyelinase-inhibited autophagy in melanoma response to cisplatin.

    PubMed

    Cervia, Davide; Assi, Emma; De Palma, Clara; Giovarelli, Matteo; Bizzozero, Laura; Pambianco, Sarah; Di Renzo, Ilaria; Zecchini, Silvia; Moscheni, Claudia; Vantaggiato, Chiara; Procacci, Patrizia; Clementi, Emilio; Perrotta, Cristiana

    2016-05-01

    The sphingolipid metabolising enzyme Acid Sphingomyelinase (A-SMase) has been recently shown to inhibit melanoma progression and correlate inversely to tumour grade. In this study we have investigated the role of A-SMase in the chemo-resistance to anticancer treatmentusing mice with melanoma allografts and melanoma cells differing in terms of expression/activity of A-SMase. Since autophagy is emerging as a key mechanism in tumour growth and chemo-resistance, we have also investigated whether an action of A-SMase in autophagy can explain its role. Melanoma sensitivity to chemotherapeutic agent cisplatin in terms of cell viability/apoptosis, tumour growth, and animal survival depended directly on the A-SMase levels in tumoural cells. A-SMase action was due to inhibition of autophagy through activation of Akt/mammalian target of rapamycin (mTOR) pathway. Treatment of melanoma-bearing mice with the autophagy inhibitor chloroquine restored sensitivity to cisplatin of tumours expressing low levels of A-SMase while no additive effects were observed in tumours characterised by sustained A-SMase levels. The fact that A-SMase in melanomas affects mTOR-regulated autophagy and plays a central role in cisplatin efficacy encourages pre-clinical testing on the modulation of A-SMase levels/activity as possible novel anti-neoplastic strategy. PMID:27107419

  6. Modulation of Acid Sphingomyelinase in Melanoma Reprogrammes the Tumour Immune Microenvironment.

    PubMed

    Assi, Emma; Cervia, Davide; Bizzozero, Laura; Capobianco, Annalisa; Pambianco, Sarah; Morisi, Federica; De Palma, Clara; Moscheni, Claudia; Pellegrino, Paolo; Clementi, Emilio; Perrotta, Cristiana

    2015-01-01

    The inflammatory microenvironment induces tumours to acquire an aggressive and immunosuppressive behaviour. Since acid sphingomyelinase (A-SMase) downregulation in melanoma was shown to determine a malignant phenotype, we aimed here to elucidate the role of A-SMase in the regulation of tumour immunogenic microenvironment using in vivo melanoma models in which A-SMase was either downregulated or maintained at constitutively high levels. We found high levels of inflammatory factors in low A-SMase expressing tumours, which also displayed an immunosuppressive/protumoural microenvironment: high levels of myeloid-derived suppressor cells (MDSCs) and regulatory T lymphocytes (Tregs), as well as low levels of dendritic cells (DCs). In contrast, the restoration of A-SMase in melanoma cells not only reduced tumour growth and immunosuppression, but also induced a high recruitment at tumour site of effector immune cells with an antitumoural function. Indeed, we observed a poor homing of MDSCs and Tregs and the increased recruitment of CD8(+) and CD4(+) T lymphocytes as well as the infiltration of DCs and CD8(+)/CD44(high) T lymphocytes. This study demonstrates that change of A-SMase expression in cancer cells is sufficient per se to tune in vivo melanoma growth and that A-SMase levels modulate immune cells at tumour site. This may be taken into consideration in the setting of therapeutic strategies. PMID:26101462

  7. Acid sphingomyelinase-ceramide system in steatohepatitis: a novel target regulating multiple pathways.

    PubMed

    Garcia-Ruiz, Carmen; Mato, Jose M; Vance, Dennis; Kaplowitz, Neil; Fernández-Checa, José C

    2015-01-01

    Steatohepatitis (SH) is an intermediate stage of fatty liver disease and is one of the most common causes of chronic liver disease worldwide that may progress to cirrhosis and liver cancer. SH encompasses alcoholic and non-alcoholic steatohepatitis, the latter being of particular concern as it is associated with obesity and insulin resistance and has become a major cause of liver transplantation. The molecular mechanisms governing the transition from steatosis to SH are not fully understood. Here we discuss emerging data indicating that the acid sphingomyelinase (ASMase), a specific mechanism of ceramide generation, is required for the activation of key pathways that regulate steatosis, fibrosis and lipotoxicity, including endoplasmic reticulum stress, autophagy and lysosomal membrane permeabilization. Moreover, ASMase modulates alterations of the methionine cycle and phosphatidylcholine homeostasis, two crucial events involved in SH that regulate methylation reactions, antioxidant defence and membrane integrity. These new findings suggest that targeting ASMase in combination with restoring methionine metabolism and phosphatidylcholine levels may be of utility in the treatment of SH. PMID:25281863

  8. Cholesterol-mediated activation of acid sphingomyelinase disrupts autophagy in the retinal pigment epithelium

    PubMed Central

    Toops, Kimberly A.; Tan, Li Xuan; Jiang, Zhichun; Radu, Roxana A.; Lakkaraju, Aparna

    2015-01-01

    Autophagy is an essential mechanism for clearing damaged organelles and proteins within the cell. As with neurodegenerative diseases, dysfunctional autophagy could contribute to blinding diseases such as macular degeneration. However, precisely how inefficient autophagy promotes retinal damage is unclear. In this study, we investigate innate mechanisms that modulate autophagy in the retinal pigment epithelium (RPE), a key site of insult in macular degeneration. High-speed live imaging of polarized adult primary RPE cells and data from a mouse model of early-onset macular degeneration identify a mechanism by which lipofuscin bisretinoids, visual cycle metabolites that progressively accumulate in the RPE, disrupt autophagy. We demonstrate that bisretinoids trap cholesterol and bis(monoacylglycero)phosphate, an acid sphingomyelinase (ASMase) cofactor, within the RPE. ASMase activation increases cellular ceramide, which promotes tubulin acetylation on stabilized microtubules. Live-imaging data show that autophagosome traffic and autophagic flux are inhibited in RPE with acetylated microtubules. Drugs that remove excess cholesterol or inhibit ASMase reverse this cascade of events and restore autophagosome motility and autophagic flux in the RPE. Because accumulation of lipofuscin bisretinoids and abnormal cholesterol homeostasis are implicated in macular degeneration, our studies suggest that ASMase could be a potential therapeutic target to ensure the efficient autophagy that maintains RPE health. PMID:25378587

  9. Structural Basis for Nucleotide Hydrolysis by the Acid Sphingomyelinase-like Phosphodiesterase SMPDL3A.

    PubMed

    Gorelik, Alexei; Illes, Katalin; Superti-Furga, Giulio; Nagar, Bhushan

    2016-03-18

    Sphingomyelin phosphodiesterase, acid-like 3A (SMPDL3A) is a member of a small family of proteins founded by the well characterized lysosomal enzyme, acid sphingomyelinase (ASMase). ASMase converts sphingomyelin into the signaling lipid, ceramide. It was recently discovered that, in contrast to ASMase, SMPDL3A is inactive against sphingomyelin and, surprisingly, can instead hydrolyze nucleoside diphosphates and triphosphates, which may play a role in purinergic signaling. As none of the ASMase-like proteins has been structurally characterized to date, the molecular basis for their substrate preferences is unknown. Here we report crystal structures of murine SMPDL3A, which represent the first structures of an ASMase-like protein. The catalytic domain consists of a central mixed β-sandwich surrounded by α-helices. Additionally, SMPDL3A possesses a unique C-terminal domain formed from a cluster of four α-helices that appears to distinguish this protein family from other phosphoesterases. We show that SMDPL3A is a di-zinc-dependent enzyme with an active site configuration that suggests a mechanism of phosphodiester hydrolysis by a metal-activated water molecule and protonation of the leaving group by a histidine residue. Co-crystal structures of SMPDL3A with AMP and α,β-methylene ADP (AMPCP) reveal that the substrate binding site accommodates nucleotides by establishing interactions with their base, sugar, and phosphate moieties, with the latter the major contributor to binding affinity. Our study provides the structural basis for SMPDL3A substrate specificity and sheds new light on the function of ASMase-like proteins. PMID:26792860

  10. Acid sphingomyelinase (aSMase) deficiency leads to abnormal microglia behavior and disturbed retinal function

    SciTech Connect

    Dannhausen, Katharina; Karlstetter, Marcus; Caramoy, Albert; Volz, Cornelia; Jägle, Herbert; Liebisch, Gerhard; Utermöhlen, Olaf; Langmann, Thomas

    2015-08-21

    Mutations in the acid sphingomyelinase (aSMase) coding gene sphingomyelin phosphodiesterase 1 (SMPD1) cause Niemann-Pick disease (NPD) type A and B. Sphingomyelin storage in cells of the mononuclear phagocyte system cause hepatosplenomegaly and severe neurodegeneration in the brain of NPD patients. However, the effects of aSMase deficiency on retinal structure and microglial behavior have not been addressed in detail yet. Here, we demonstrate that retinas of aSMase{sup −/−} mice did not display overt neuronal degeneration but showed significantly reduced scotopic and photopic responses in electroretinography. In vivo fundus imaging of aSMase{sup −/−} mice showed many hyperreflective spots and staining for the retinal microglia marker Iba1 revealed massive proliferation of retinal microglia that had significantly enlarged somata. Nile red staining detected prominent phospholipid inclusions in microglia and lipid analysis showed significantly increased sphingomyelin levels in retinas of aSMase{sup −/−} mice. In conclusion, the aSMase-deficient mouse is the first example in which microglial lipid inclusions are directly related to a loss of retinal function. - Highlights: • aSMase-deficient mice show impaired retinal function and reactive microgliosis. • aSMase-deficient microglia express pro-inflammatory transcripts. • aSMase-deficient microglia proliferate and have increased cell body size. • In vivo imaging shows hyperreflective spots in the fundus of aSMase-deficient mice. • aSMase-deficient microglia accumulate sphingolipid-rich intracellular deposits.

  11. Propionibacterium acnes CAMP Factor and Host Acid Sphingomyelinase Contribute to Bacterial Virulence: Potential Targets for Inflammatory Acne Treatment

    PubMed Central

    Nakatsuji, Teruaki; Tang, De-chu C.; Zhang, Liangfang; Gallo, Richard L.; Huang, Chun-Ming

    2011-01-01

    Background In the progression of acne vulgaris, the disruption of follicular epithelia by an over-growth of Propionibacterium acnes (P. acnes) permits the bacteria to spread and become in contact with various skin and immune cells. Methodology/Principal Findings We have demonstrated in the present study that the Christie, Atkins, Munch-Peterson (CAMP) factor of P. acnes is a secretory protein with co-hemolytic activity with sphingomyelinase that can confer cytotoxicity to HaCaT keratinocytes and RAW264.7 macrophages. The CAMP factor from bacteria and acid sphingomyelinase (ASMase) from the host cells were simultaneously present in the culture supernatant only when the cells were co-cultured with P. acnes. Either anti-CAMP factor serum or desipramine, a selective ASMase inhibitor, significantly abrogated the P. acnes-induced cell death of HaCaT and RAW264.7 cells. Intradermal injection of ICR mouse ears with live P. acnes induced considerable ear inflammation, macrophage infiltration, and an increase in cellular soluble ASMase. Suppression of ASMase by systemic treatment with desipramine significantly reduced inflammatory reaction induced by intradermal injection with P. acnes, suggesting the contribution of host ASMase in P. acnes-induced inflammatory reaction in vivo. Vaccination of mice with CAMP factor elicited a protective immunity against P. acnes-induced ear inflammation, indicating the involvement of CAMP factor in P. acnes-induced inflammation. Most notably, suppression of both bacterial CAMP factor and host ASMase using vaccination and specific antibody injection, respectively, cooperatively alleviated P. acnes-induced inflammation. Conclusions/Significance These findings envision a novel infectious mechanism by which P. acnes CAMP factor may hijack host ASMase to amplify bacterial virulence to degrade and invade host cells. This work has identified both CAMP factor and ASMase as potential molecular targets for the development of drugs and vaccines against

  12. Role of acid sphingomyelinase in the age-dependent dysregulation of sphingolipids turnover in the tissues of rats.

    PubMed

    Babenko, Nataliya A; Garkavenko, Vladimir V; Storozhenko, Galina V; Timofiychuk, Olga A

    2016-04-01

    Old age-associated pathologies usually coincide with altered sphingolipid metabolism. In the present article, the role of acid sphingomyelinase (ASMase) in the age-dependent changes of sphingomyelin (SM) and ceramide contents in the tissues has been investigated by means of ASMase inhibitors, imipramine and zoledronic acid. It has been determined that ceramide content and ceramide/SM ratio increased, while SM level decreased in the heart, liver, blood serum and skeletal muscles of 24-month old rats in contrast to 3-month old animals. Injections of imipramine or zoledronic acid to 24-month old rats resulted in significant downregulation of ASMase in the liver and skeletal and heart muscles. The both inhibitors decreased the ceramide content and ceramide/SM ratio and increased the SM content in all tissues studied, except the heart, of old rats to the levels close to those observed in the young animals. Long-term treatment of rats by inhibitors, which have different mechanisms of action on ASMase, exerts the similar, but not equal effects on enzyme activity and SM turnover. In summary, the data above strongly suggest that the age-dependent up-regulation of ASMase plays an important role in the modulation of ceramide and SM contents in rat tissues and that imipramine and zoledronic acid are useful tools for SM turnover manipulation at old age. PMID:26830134

  13. Caspase-8 and caspase-7 sequentially mediate proteolytic activation of acid sphingomyelinase in TNF-R1 receptosomes

    PubMed Central

    Edelmann, Bärbel; Bertsch, Uwe; Tchikov, Vladimir; Winoto-Morbach, Supandi; Perrotta, Cristiana; Jakob, Marten; Adam-Klages, Sabine; Kabelitz, Dieter; Schütze, Stefan

    2011-01-01

    We previously demonstrated that tumour necrosis factor (TNF)-induced ceramide production by endosomal acid sphingomyelinase (A-SMase) couples to apoptosis signalling via activation of cathepsin D and cleavage of Bid, resulting in caspase-9 and caspase-3 activation. The mechanism of TNF-mediated A-SMase activation within the endolysosomal compartment is poorly defined. Here, we show that TNF-induced A-SMase activation depends on functional caspase-8 and caspase-7 expression. The active forms of all three enzymes, caspase-8, caspase-7 and A-SMase, but not caspase-3, colocalize in internalized TNF receptosomes. While caspase-8 and caspase-3 are unable to induce activation of purified pro-A-SMase, we found that caspase-7 mediates A-SMase activation by direct interaction resulting in proteolytic cleavage of the 72-kDa pro-A-SMase zymogen at the non-canonical cleavage site after aspartate 253, generating an active 57 kDa A-SMase molecule. Caspase-7 down modulation revealed the functional link between caspase-7 and A-SMase, confirming proteolytic cleavage as one further mode of A-SMase activation. Our data suggest a signalling cascade within TNF receptosomes involving sequential activation of caspase-8 and caspase-7 for induction of A-SMase activation by proteolytic cleavage of pro-A-SMase. PMID:21157428

  14. A novel cisplatin mediated apoptosis pathway is associated with acid sphingomyelinase and FAS proapoptotic protein activation in ovarian cancer.

    PubMed

    Maurmann, L; Belkacemi, L; Adams, N R; Majmudar, P M; Moghaddas, S; Bose, R N

    2015-07-01

    Platinum-based anticancer drugs, including cisplatin and carboplatin, have been cornerstones in the treatment of solid tumors. We report here that these DNA-damaging agents, particularly cisplatin, induce apoptosis through plasma membrane disruption, triggering FAS death receptor via mitochondrial (intrinsic) pathways. Our objectives were to: quantify the composition of membrane metabolites; and determine the potential involvement of acid sphingomyelinase (ASMase) in the FAS-mediated apoptosis in ovarian cancer after cisplatin treatment. The resulting analysis revealed enhanced apoptosis as measured by: increased phosphocholine, and glycerophosphocholine; elevated cellular energetics; and phosphocreatine and nucleoside triphosphate concentrations. The plasma membrane alterations were accompanied by increased ASMase activity, leading to the upregulation of FAS, FASL and related pro-apoptotic BAX and PUMA genes. Moreover FAS, FASL, BAX, PUMA, CASPASE-3 and -9 proteins were upregulated. Our findings implicate ASMase activity and the intrinsic pathways in cisplatin-mediated membrane demise, and contribute to our understanding of the mechanisms by which ovarian tumors may become resistant to cisplatin. PMID:25846011

  15. Acid sphingomyelinase determines melanoma progression and metastatic behaviour via the microphtalmia-associated transcription factor signalling pathway

    PubMed Central

    Bizzozero, L; Cazzato, D; Cervia, D; Assi, E; Simbari, F; Pagni, F; De Palma, C; Monno, A; Verdelli, C; Querini, P R; Russo, V; Clementi, E; Perrotta, C

    2014-01-01

    Melanoma is a rapidly growing and highly metastatic cancer with high mortality rates, for which a resolutive treatment is lacking. Identification of novel therapeutic strategies and biomarkers of tumour stage is thus of particular relevance. We report here on a novel biomarker and possible candidate therapeutic target, the sphingolipid metabolising enzyme acid sphingomyelinase (A-SMase). A-SMase expression correlates inversely with tumour stage in human melanoma biopsies. Studies in a mouse model of melanoma and on cell lines derived from mouse and human melanomas demonstrated that A-SMase levels of expression actually determine the malignant phenotype of melanoma cells in terms of pigmentation, tumour progression, invasiveness and metastatic ability. The action of A-SMase is mediated by the activation of the extracellular signal-regulated kinase, the subsequent proteasomal degradation of the Microphtalmia-associated transcription factor (Mitf) and inhibition of cyclin-dependent kinase 2, Bcl-2 and c-Met, downstream targets of Mitf involved in tumour cell proliferation, survival and metastatisation. PMID:24317198

  16. A new fluorimetric enzyme assay for the diagnosis of Niemann-Pick A/B, with specificity of natural sphingomyelinase substrate.

    PubMed

    van Diggelen, O P; Voznyi, Ya V; Keulemans, J L M; Schoonderwoerd, K; Ledvinova, J; Mengel, E; Zschiesche, M; Santer, R; Harzer, K

    2005-01-01

    6-Hexadecanoylamino-4-methylumbelliferylphosphorylcholine (HMUPC) was shown to be a specific substrate for the determination of acid (lysosomal) sphingomyelinase (ASM; gene SMPD1). Fibroblasts (n = 27) and leukocytes (n = 8) from both the A and B types of Niemann-Pick disease showed < 6% and < 10% of mean normal ASM activity, respectively. Niemann-Pick A or B patients bearing the Q292K mutation had apparently normal ASM activity with our new artificial substrate. These patients with false-normal sphingomyelinase activity, however, could readily be detected by determining the extent of inhibition of enzymatic hydrolysis of the artificial substrate HMU-PC by an unlabelled natural substrate, in particular lysosphingomyelin. This approach is generally applicable. Our novel assay for ASM combines the ease of a rapid and robust enzyme assay using a fluorogenic substrate with the specificity of an ASM assay using a natural substrate. Such assays are obviously more convenient to the diagnostic laboratory, since radiolabelled substrates are not required. PMID:16151905

  17. Regulation of dynein-mediated autophagosomes trafficking by ASM in CASMCs

    PubMed Central

    Li, Pin-Lan; Nguyen, Thaison; Li, Xiang; Zhang, Yang

    2016-01-01

    Acid sphingomyelinase (ASM; gene symbol Smpd1) has been shown to play a crucial role in autophagy maturation by controlling lysosomal fusion with autophagosomes in coronary arterial smooth muscle cells (CASMCs). However, the underlying molecular mechanism by which ASM controls autophagolysosomal fusion remains unknown. In primary cultured CASMCs, lysosomal Ca2+ induced by 7-ketocholesterol (7-Ket, an atherogenic stimulus and autophagy inducer) was markedly attenuated by ASM deficiency or TRPML1 gene silencing suggesting that ASM signaling is required for TRPML1 channel activity and subsequent lysosomal Ca2+ release. In these CASMCs, ASM deficiency or TRPML1 gene silencing markedly inhibited 7-Ket-induced dynein activation. In addition, 7-Ket-induced autophagosome trafficking, an event associated with lysosomal Ca2+ release and dynein activity, was significantly inhibited in ASM-deficient (Smpd1−/−) CASMCs compared to that in Smpd1+/+ CASMCs. Finally, overexpression of TRPML1 proteins restored 7-Ket-induced lysosomal Ca2+ release and autophagosome trafficking in Smpd1−/− CASMCs. Collectively, these results suggest that ASM plays a critical role in regulating lysosomal TRPML1-Ca2+ signaling and subsequent dynein-mediated autophagosome trafficking, which leads its role in controlling autophagy maturation in CASMCs under atherogenic stimulation. PMID:26709800

  18. Neutral Sphingomyelinase 2

    PubMed Central

    Filosto, Simone; Castillo, Sianna; Danielson, Aaron; Franzi, Lisa; Khan, Elaine; Kenyon, Nick; Last, Jerold; Pinkerton, Kent; Tuder, Rubin; Goldkorn, Tzipora

    2011-01-01

    Chronic obstructive pulmonary disease (COPD) is caused by exposure to cigarette smoke (CS). One mechanism of CS-induced lung injury is aberrant generation of ceramide, which leads to elevated apoptosis of epithelial and endothelial cells in the alveolar spaces. Recently, we discovered that CS-induced ceramide generation and apoptosis in pulmonary cells is governed by neutral sphingomyelinase (nSMase) 2. In the current experiments, we expanded our studies to investigate whether nSMase2 governs ceramide generation and apoptosis in vivo using rodent and human models of CS-induced lung injury. We found that exposure of mice or rats to CS leads to colocalizing elevations of ceramide levels and terminal deoxynucleotidyl transferase mediated X-dUTP nick end labeling–positive cells in lung tissues. These increases are nSMase2 dependent, and are abrogated by treatment with N-acetyl cysteine or anti-nSMase2 small interfering RNA (siRNA). We further showed that mice that are heterozygous for nSMase2 demonstrate significant decrease in ceramide generation after CS exposure, whereas acidic sphingomyelinase (aSMase) knockout mice maintain wild-type ceramide levels, confirming our previous findings (in human airway epithelial cells) that only nSMase2, and not aSMase, is activated by CS exposure. Lastly, we found that lung tissues from patients with emphysema (smokers) display significantly higher levels of nSMase2 expression compared with lung tissues from healthy control subjects. Taken together, these data establish the central in vivo role of nSMase2 in ceramide generation, aberrant apoptosis, and lung injury under CS exposure, underscoring its promise as a novel target for the prevention of CS-induced airspace destruction. PMID:20448054

  19. [Structure and function of sphingomyelinase].

    PubMed

    Oda, Masataka

    2009-10-01

    Bacillus cereus is one that causes of opportunistic human infections. Sphingomyelinase produced by B. cereus is assumed a virulence factor for the infection. Sphingomyelinase from Bacillus cereus (Bc-SMase) is Mg(2+)-containing metalloenzyme. Bc-SMase is a family of neutral SMase (nSMase) and mimics the actions of the endogenous mammalian nSMase in causing differentiation, development, and apoptosis. Bc-SMase may be a good model for the poorly characterized mammalian nSMase. Activation of Bc-SMase by divalent metal ions was in the order Co(2+)>Mn(2+)>Mg(2+)>Ca(2+)>Sr(2+). Crystal structure analysis of Bc-SMase bound to Co(2+), Mg(2+), or Ca(2+) revealed that the water-bridged double divalent metal ions at the center of the cleft in both the Co(2+)- and Mg(2+)-bound forms is the catalytic architecture required for sphingomyelinase activity. In contrast, the architecture of Ca(2+) binding at the site showed only one binding site. A further single metal-binding site existed at one side edge of the cleft. Based on the highly conserved nature of amino acid residues of the binding sites, the crystal structure of Bc-SMase with Mg(2+) or Co(2+) provided a common structural framework applicable to phosphohydrolases belonging to the DNase I-like folding superfamily. In addition, our analysis provided evidence that beta-hairpin containing the aromatic amino acid residues and the metal ion of the side-edge participate in binding to sphinogmyelin and membranes containing sphingomyelin. This article summarized current knowledge of characteristics and mode of action of Bc-SMase. PMID:19797879

  20. Identification and Biochemical Characterization of an Acid Sphingomyelinase-Like Protein from the Bacterial Plant Pathogen Ralstonia solanacearum that Hydrolyzes ATP to AMP but Not Sphingomyelin to Ceramide

    PubMed Central

    Airola, Michael V.; Tumolo, Jessica M.; Snider, Justin; Hannun, Yusuf A.

    2014-01-01

    Acid sphingomyelinase (aSMase) is a human enzyme that catalyzes the hydrolysis of sphingomyelin to generate the bioactive lipid ceramide and phosphocholine. ASMase deficiency is the underlying cause of the genetic diseases Niemann-Pick Type A and B and has been implicated in the onset and progression of a number of other human diseases including cancer, depression, liver, and cardiovascular disease. ASMase is the founding member of the aSMase protein superfamily, which is a subset of the metallophosphatase (MPP) superfamily. To date, MPPs that share sequence homology with aSMase, termed aSMase-like proteins, have been annotated and presumed to function as aSMases. However, none of these aSMase-like proteins have been biochemically characterized to verify this. Here we identify RsASML, previously annotated as RSp1609: acid sphingomyelinase-like phosphodiesterase, as the first bacterial aSMase-like protein from the deadly plant pathogen Ralstonia solanacearum based on sequence homology with the catalytic and C-terminal domains of human aSMase. A biochemical characterization of RsASML does not support a role in sphingomyelin hydrolysis but rather finds RsASML capable of acting as an ATP diphosphohydrolase, catalyzing the hydrolysis of ATP and ADP to AMP. In addition, RsASML displays a neutral, not acidic, pH optimum and prefers Ni2+ or Mn2+, not Zn2+, for catalysis. This alters the expectation that all aSMase-like proteins function as acid SMases and expands the substrate possibilities of this protein superfamily to include nucleotides. Overall, we conclude that sequence homology with human aSMase is not sufficient to predict substrate specificity, pH optimum for catalysis, or metal dependence. This may have implications to the biochemically uncharacterized human aSMase paralogs, aSMase-like 3a (aSML3a) and aSML3b, which have been implicated in cancer and kidney disease, respectively, and assumed to function as aSMases. PMID:25144372

  1. Hexachlorobenzene-induced alterations on neutral and acidic sphingomyelinases and serine palmitoyltransferase activities. A time course study in two strains of rats.

    PubMed

    Billi de Catabbi, S C; Setton-Advruj, C P; Sterin-Speziale, N; San Martín de Viale, L C; Cochón, A C

    2000-08-21

    Hexachlorobenzene (HCB) induces porphyria both in humans and rodents, and hepatocarcinoma in rodents. In a previous work we observed that HCB produces a continuous decrease in hepatic sphingomyelin (SM) content in Wistar rats. A distinguishing characteristic of sphingolipids breakdown products is their participation in anti-proliferative and apoptotic processes and in the suppression of oncogenesis. As a first step to elucidate the role of SM decrease in the hepatotoxicity induced by HCB, the present study evaluates the metabolic causes of the continuous decrease in hepatic SM content observed in Wistar rats with HCB intoxication, and its relation with porphyria development. For this purpose, the time-course (3, 7, 15, 21 and 28 days) of the effects of HCB on hepatic SM levels and on some of the enzymes of SM synthesis (serine palmitoyltransferase, SPT) and catabolism (sphingomyelinases, SMases) was followed, using two strains of rats differing in their susceptibility to acquire porphyria: Chbb THOM (low) and Wistar (high). HCB (1 g kg(-1) b.w. per day) was administered by gastric intubation as an aqueous suspension. After 5 days of HCB treatment, animals were allowed a 2-day recovery period without HCB administration. Two phases in the HCB-induced damages to sphingolipid metabolism were observed. The first stage (7 days of treatment), common to both strains of rats, was characterized by a decrease in hepatic SM levels (17-25%) and in SPT activity (50-43%), while strain differences were found for the later stage. In Chbb THOM rats, hepatic SM content was restored to normal values concomitantly with an increase in SPT activity (44%, at day 28), and without any increase in SM catabolism. In addition, the level of the other phospholipids was not altered. In Wistar rats, hepatic SM levels decreased continuously throughout the experiment, accompanied by increases in SPT, acidic sphingomyelinase (A-SMase) and neutral sphingomyelinase (N-SMase) activities (86, 28.5 and 78

  2. Lysosomal sphingomyelinase is not solicited for apoptosis signaling.

    PubMed

    Bezombes, C; Ségui, B; Cuvillier, O; Bruno, A P; Uro-Coste, E; Gouazé, V; Andrieu-Abadie, N; Carpentier, S; Laurent, G; Salvayre, R; Jaffrézou, J P; Levade, T

    2001-02-01

    Stress-induced activation of an acidic sphingomyelinase leading to generation of ceramide, an important lipid mediator, has been associated with apoptosis; however, the implication of this hydrolase has been questioned. The present study aimed at re-evaluating the role of this lysosomal enzyme in apoptosis initiated by different apoptotic inducers. The sensitivity of a series of acid sphingomyelinase-deficient cell lines derived from Niemann-Pick disease patients to stress-induced apoptosis was investigated. We have now shown that stress stimuli, such as anthracyclines, ionizing radiation, and Fas ligation trigger similar apoptotic hallmarks in normal and acid sphingomyelinase-deficient cell lines. Retrovirus-mediated gene correction of enzyme deficiency in Niemann-Pick cells does not modify response to apoptosis. Ceramide production is comparable in normal and Niemann-Pick cells, and increased activity of neutral sphingomyelinase is observed. Thus, our findings cast serious doubts that lysosomal sphingomyelinase activation is responsible for stress-induced apoptosis of cultured cells. PMID:11156942

  3. Molecular modeling of human alkaline sphingomyelinase.

    PubMed

    Suresh, Panneer Selvam; Olubiyi, Olujide; Thirunavukkarasu, Chinnasamy; Strodel, Birgit; Kumar, Muthuvel Suresh

    2011-01-01

    Alkaline sphingomyelinase, which is expressed in the human intestine and hydrolyses sphingomyelin, is a component of the plasma and the lysosomal membranes. Hydrolase of sphingomyelin generates ceramide, sphingosine, and sphingosine 1-phosphate that have regulatory effects on vital cellular functions such as proliferation, differentiation, and apoptosis. The enzyme belongs to the Nucleotide Pyrophosphatase/Phosphodiesterase family and it differs in structural similarity with acidic and neutral sphingomyelinase. In the present study we modeled alkaline sphingomyelinase using homology modeling based on the structure of Nucleotide Pyrophosphatase/Phosphodiesterase from Xanthomonas axonopodis with which it shares 34% identity. Homology modeling was performed using Modeller9v7. We found that Cys78 and Cys394 form a disulphide bond. Further analysis shows that Ser76 may be important for the function of this enzyme, which is supported by the findings of Wu et al. (2005), that S76F abolishes the activity completely. We found that the residues bound to Zn(2+) are conserved and geometrically similar with the template. Molecular Dynamics simulations were carried out for the modeled protein to observe the effect of Zinc metal ions. It was observed that the metal ion has little effect with regard to the stability but induces increased fluctuations in the protein. These analyses showed that Zinc ions play an important role in stabilizing the secondary structure and in maintaining the compactness of the active site. PMID:21544170

  4. TRAIL-Death Receptor 4 Signaling via Lysosome Fusion and Membrane Raft Clustering In Coronary Arterial Endothelial Cells: Evidence from ASM Knockout Mice

    PubMed Central

    Li, Xiang; Han, Wei-Qing; Boini, Krishna M.; Xia, Min; Zhang, Yang; Li, Pin-Lan

    2012-01-01

    Tumor necrosis factor (TNF)-related apoptosis inducing ligand (TRAIL) and its receptor death receptor 4 (DR4) have been implicated in the development of endothelial dysfunction and atherosclerosis. However, the signaling mechanism mediating DR4 activation and leading to endothelial injury remains unclear. We recently demonstrated that ceramide production via hydrolysis of membrane sphingomyelin by acid sphingomyelinase (ASM) results in membrane raft (MRs) clustering and formation of important redox signaling platforms, which play a crucial role in amplifying redox signaling in endothelial cells leading to endothelial dysfunction. The present study aims to investigate whether TRAIL triggers MR clustering via lysosome fusion and ASM activation, thereby conducting transmembrane redox signaling and changing endothelial function. Using confocal microscopy, we found that TRAIL induced MR clustering and its co-localization with DR4 in coronary arterial endothelial cells (CAECs) isolated from wild-type (Smpd1+/+) mice. Further, TRAIL triggered ASM translocation, ceramide production and NADPH oxidase aggregation in MR clusters in Smpd1+/+ CAECs, whereas these observations were not found in Smpd1−/− CAECs. Moreover, ASM deficiency reduced TRAIL-induced O2−· production in CAECs and abolished TRAIL-induced impairment on endothelium-dependent vasodilation in small resistance arteries. By measuring fluorescence resonance energy transfer (FRET), we found that Lamp-1 (lysosome membrane marker protein) and ganglioside GM1 (MR marker) were trafficking together in Smpd1+/+ CAECs, which was absent in Smpd1−/− CAECs. Consistently, fluorescence imaging of living cells with specific lysosome probes demonstrated that TRAIL-induced lysosome fusion with membrane was also absent in Smpd1−/− CAECs. Taken together, these results suggest that ASM is essential for TRAIL-induced lysosomal trafficking and fusion with membrane and formation of MR redox signaling platforms, which may

  5. Synthesis of substrates for periodate-coupled assay of phospholipases C and sphingomyelinases.

    PubMed

    Larsen, Kira Løw; Andersen, Rokhsana J; Brask, Jesper

    2016-09-01

    A series of 4-nitrophenyl (pNP) and 4-methylumbelliferyl (4MU) substrate analogues of phosphatidyl choline (PC) and phosphatidic acid (PA) were synthesized from 4-bromo-1-butene by ether formation, olefin epoxidation and ring opening with the phosphate head group. The pNP PC analogue, 4-(4-nitrophenoxy)-2-hydroxy-butyl-1-phosphoryl choline (1) was evaluated in assays of fungal sphingomyelinases, also displaying phospholipase C activity. Reactions were terminated with a periodate-containing stop solution, leading to liberation of pNP, quantified spectrophotometrically in an end-point measurement. A kinetic evaluation of sphingomyelinases from Kionochaeta sp. and Penicillium emersonii showed relatively high KM and low kcat values for this substrate, limiting its practical applicability in assays with low sphingomyelinase concentrations. PMID:27444331

  6. Generation of biologically active linear and cyclic peptides has revealed a unique fine specificity of rituximab and its possible cross-reactivity with acid sphingomyelinase-like phosphodiesterase 3b precursor.

    PubMed

    Perosa, Federico; Favoino, Elvira; Caragnano, Maria Antonietta; Dammacco, Franco

    2006-02-01

    Heterogeneity of the effector functions displayed by rituximab and other anti-CD20 monoclonal antibodies (mAbs) apparently recognizing the same CD20 epitope suggests that additional mechanisms, probably related to mAb fine specificity, are responsible for B-cell depletion. To improve our understanding of rituximab's function, its fine specificity was investigated by means of phage display peptide library (PDPL)-expressing 7-mer cyclic (c7c) or 7-/12-mer linear peptides. Rituximab-specific c7c PDPL-derived clone insert sequences expressed the motif A(S)NPS overlapping the human CD20 170ANPS173. P172 was the most critical for rituximab binding, since its replacement with S172 (of mouse CD20) abolished the reactivity. The WPXWLE motif expressed by the linear PDPL-derived clone insert sequences could only be aligned to the reverse-oriented 161WPXWLE156 of acid sphingomyelinase-like phosphodiesterase 3b precursor (ASMLPD), though linear peptides bearing WPXWLE competed with cyclic ones for rituximab-paratope binding. Anti-CD20 mAb 1F5 only displayed a reactivity profile similar to that of rituximab, which also reacted with ASMLPD-derived peptides. Peptides induced antibodies with specificity and effector functions similar to those of rituximab. Our results show a unique fine specificity of rituximab, define the molecular basis for the lack of rituximab reactivity with mouse CD20 (mCD20), and the potential of targeting CD20 in an active immunotherapy setting. A possible rituximab interaction with ASMLPD is suggested. PMID:16223774

  7. Bacterial Sphingomyelinases and Phospholipases as Virulence Factors.

    PubMed

    Flores-Díaz, Marietta; Monturiol-Gross, Laura; Naylor, Claire; Alape-Girón, Alberto; Flieger, Antje

    2016-09-01

    Bacterial sphingomyelinases and phospholipases are a heterogeneous group of esterases which are usually surface associated or secreted by a wide variety of Gram-positive and Gram-negative bacteria. These enzymes hydrolyze sphingomyelin and glycerophospholipids, respectively, generating products identical to the ones produced by eukaryotic enzymes which play crucial roles in distinct physiological processes, including membrane dynamics, cellular signaling, migration, growth, and death. Several bacterial sphingomyelinases and phospholipases are essential for virulence of extracellular, facultative, or obligate intracellular pathogens, as these enzymes contribute to phagosomal escape or phagosomal maturation avoidance, favoring tissue colonization, infection establishment and progression, or immune response evasion. This work presents a classification proposal for bacterial sphingomyelinases and phospholipases that considers not only their enzymatic activities but also their structural aspects. An overview of the main physiopathological activities is provided for each enzyme type, as are examples in which inactivation of a sphingomyelinase- or a phospholipase-encoding gene impairs the virulence of a pathogen. The identification of sphingomyelinases and phospholipases important for bacterial pathogenesis and the development of inhibitors for these enzymes could generate candidate vaccines and therapeutic agents, which will diminish the impacts of the associated human and animal diseases. PMID:27307578

  8. A novel mitochondrial sphingomyelinase in zebrafish cells.

    PubMed

    Yabu, Takeshi; Shimuzu, Akio; Yamashita, Michiaki

    2009-07-24

    Sphingolipids are important signaling molecules in many biological processes, but little is known regarding their physiological roles in the mitochondrion. We focused on the biochemical characters of a novel sphingomyelinase (SMase) and its function in mitochondrial ceramide generation in zebrafish embryonic cells. The cloned SMase cDNA encoded a polypeptide of 545 amino acid residues (putative molecular weight, 61,300) containing a mitochondrial localization signal (MLS) and a predicted transmembrane domain. The mature endogenous enzyme was predicted to have a molecular weight of 57,000, and matrix-assisted laser de sorption ionization time-of-flight mass spectrometry analysis indicated that the N-terminal amino acid residue of the mature enzyme was Ala-36. The purified enzyme optimally hydrolyzed [(14)C]sphingomyelin in the presence of 10 mm Mg(2+) at pH 7.5. In HEK293 cells that overexpressed SMase cDNA, the enzyme was localized to the mitochondrial fraction, whereas mutant proteins lacking MLS or both the MLS and the transmembrane domain were absent from the mitochondrial fraction. Endogenous SMase protein co-localized with a mitochondrial cytostaining marker. Using a protease protection assay, we found that SMase was distributed throughout the intermembrane space and/or the inner membrane of the mitochondrion. Furthermore, the overexpression of SMase in HEK293 cells induced ceramide generation and sphingomyelin hydrolysis in the mitochondrial fraction. Antisense phosphorothioate oligonucleotide-induced knockdown repressed ceramide generation and sphingomyelin hydrolysis in the mitochondrial fraction in zebrafish embryonic cells. These observations indicate that SMase catalyzes the hydrolysis of sphingomyelin and generates ceramide in mitochondria in fish cells. PMID:19429680

  9. Characterization of secretory sphingomyelinase activity, lipoprotein sphingolipid content and LDL aggregation in ldlr−/− mice fed on a high-fat diet

    PubMed Central

    Deevska, Gergana M.; Sunkara, Manjula; Morris, Andrew J.; Nikolova-Karakashian, Mariana N.

    2012-01-01

    The propensity of LDLs (low-density lipoproteins) for aggregation and/or oxidation has been linked to their sphingolipid content, specifically the levels of SM (sphingomyelin) and ceramide. To investigate this association in vivo, ldlr (LDL receptor)-null mice (ldlr−/−) were fed on a modified (atherogenic) diet containing saturated fats and cholesterol. The diet led to significantly elevated SM content in all serum lipoproteins. In contrast, ceramide increased only in the LDL particles. MS-based analyses of the lipid acyl chain composition revealed a marked elevation in C16:0 fatty acid in SM and ceramide, consistent with the prevalence of palmitic acid in the modified diet. The diet also led to increased activity of the S-SMase [secretory SMase (sphingomyelinase)], a protein that is generated by ASMase (acid SMase) and acts on serum LDL. An increased macrophage secretion seemed to be responsible for the elevated S-SMase activity. ASMase-deficient mice (asm−/−/ldlr−/−) lacked S-SMase activity and were protected from diet-induced elevation in LDL ceramide. LDL from asm−/−/ldlr−/− mice fed on the modified diet were less aggregated and oxidized than LDL from asm+/+/ldlr−/− mice. When tested in vitro, the propensity for aggregation was dependent on the SM level: only LDL from animals on modified diet that have high SM content aggregated when treated with recombinant S-SMase. In conclusion, LDL-SM content and S-SMase activity are up-regulated in mice fed on an atherogenic diet. S-SMase mediates diet-induced changes in LDL ceramide content and aggregation. S-SMase effectiveness in inducing aggregation is dependent on diet-induced enrichment of LDL with SM, possibly through increased hepatic synthesis. PMID:22712892

  10. Neutral sphingomyelinase and breast cancer research.

    PubMed

    Hwang, Sun-Yong; Kim, Tae-Hee; Lee, Hae-Hyeog

    2015-04-01

    Our understanding of the functions of neutral sphingomyelinase (N-SMase) signaling has advanced over the past decade. In this review, we focus on the roles and regulation of N-SMase 1, N-SMase 2, N-SMase 3, an enzyme that generates the bioactive lipid ceramide through the hydrolysis of the membrane lipid sphingomyelin. A large body of work has now implicated N-SMase 2 in a diverse set of cellular functions, physiological processes, and disease pathologies. We focus on different aspects of this enzyme's regulation from transcriptional, post-translational, and biochemical. Furthermore, we expected N-SMase involvement in cellular processes including inflammatory signaling, cell growth, apoptosis, and tumor necrosis factor which in turn play important roles in pathologies such as cancer metastasis, variable disease, and other organ system disorders. Lastly, we examine avenues where targeted N-SMase inhibition may be clinically beneficial in disease scenarios. PMID:26046034

  11. Review and Application of ASME NOG-1 and ASME NUM-1-2000

    NASA Technical Reports Server (NTRS)

    Lytle, Bradford P.; Delgado, H. (Technical Monitor)

    2002-01-01

    The intent of the workshop is to review the application of the ASME Nuclear Crane Standards ASME NOG-1 and ASME NUM-1-2000. The ASME Nuclear Crane standards provide a basis for purchasing overhead handling equipment with enhanced safety features, based upon accepted engineering principles, and including performance and environmental parameters specific to nuclear facilities.

  12. ASME Code Efforts Supporting HTGRs

    SciTech Connect

    D.K. Morton

    2010-09-01

    In 1999, an international collaborative initiative for the development of advanced (Generation IV) reactors was started. The idea behind this effort was to bring nuclear energy closer to the needs of sustainability, to increase proliferation resistance, and to support concepts able to produce energy (both electricity and process heat) at competitive costs. The U.S. Department of Energy has supported this effort by pursuing the development of the Next Generation Nuclear Plant, a high temperature gas-cooled reactor. This support has included research and development of pertinent data, initial regulatory discussions, and engineering support of various codes and standards development. This report discusses the various applicable American Society of Mechanical Engineers (ASME) codes and standards that are being developed to support these high temperature gascooled reactors during construction and operation. ASME is aggressively pursuing these codes and standards to support an international effort to build the next generation of advanced reactors so that all can benefit.

  13. ASME Code Efforts Supporting HTGRs

    SciTech Connect

    D.K. Morton

    2011-09-01

    In 1999, an international collaborative initiative for the development of advanced (Generation IV) reactors was started. The idea behind this effort was to bring nuclear energy closer to the needs of sustainability, to increase proliferation resistance, and to support concepts able to produce energy (both electricity and process heat) at competitive costs. The U.S. Department of Energy has supported this effort by pursuing the development of the Next Generation Nuclear Plant, a high temperature gas-cooled reactor. This support has included research and development of pertinent data, initial regulatory discussions, and engineering support of various codes and standards development. This report discusses the various applicable American Society of Mechanical Engineers (ASME) codes and standards that are being developed to support these high temperature gascooled reactors during construction and operation. ASME is aggressively pursuing these codes and standards to support an international effort to build the next generation of advanced reactors so that all can benefit.

  14. ASME Code Efforts Supporting HTGRs

    SciTech Connect

    D.K. Morton

    2012-09-01

    In 1999, an international collaborative initiative for the development of advanced (Generation IV) reactors was started. The idea behind this effort was to bring nuclear energy closer to the needs of sustainability, to increase proliferation resistance, and to support concepts able to produce energy (both electricity and process heat) at competitive costs. The U.S. Department of Energy has supported this effort by pursuing the development of the Next Generation Nuclear Plant, a high temperature gas-cooled reactor. This support has included research and development of pertinent data, initial regulatory discussions, and engineering support of various codes and standards development. This report discusses the various applicable American Society of Mechanical Engineers (ASME) codes and standards that are being developed to support these high temperature gascooled reactors during construction and operation. ASME is aggressively pursuing these codes and standards to support an international effort to build the next generation of advanced reactors so that all can benefit.

  15. Experience and extensions to the ASM2 family of models.

    PubMed

    Dudley, J; Buck, G; Ashley, R; Jack, A

    2002-01-01

    The development of ASM2 has created a complex model for biological phosphorus removal. Most of the published work on calibrating this model has focused on the design of experiments to maximise information with which to calibrate the model, or the use of hourly data collected around and within an aeration tank. But many sewage works do not collect such data, nor have such instrumentation. The application of ASM2 with sparse data collected at a low frequency, and mostly only input-output, is considered in this paper, based on data collected at a Swedish sewage works. This paper shows that ASM2 can be calibrated with such measurements. This paper also looks at a modification to ASM2d to better handle heterotrophic usage of volatile fatty acids, and the use of this model to study the effects of large increases in in-sewer storage on sewage treatment works. Concern about the generation of large quantities of VFAs, and their effect on the sewage treatment processes, was unfounded. PMID:11989871

  16. Risk based ASME Code requirements

    SciTech Connect

    Gore, B.F.; Vo, T.V.; Balkey, K.R.

    1992-09-01

    The objective of this ASME Research Task Force is to develop and to apply a methodology for incorporating quantitative risk analysis techniques into the definition of in-service inspection (ISI) programs for a wide range of industrial applications. An additional objective, directed towards the field of nuclear power generation, is ultimately to develop a recommendation for comprehensive revisions to the ISI requirements of Section XI of the ASME Boiler and Pressure Vessel Code. This will require development of a firm technical basis for such requirements, which does not presently exist. Several years of additional research will be required before this can be accomplished. A general methodology suitable for application to any industry has been defined and published. It has recently been refined and further developed during application to the field of nuclear power generation. In the nuclear application probabilistic risk assessment (PRA) techniques and information have been incorporated. With additional analysis, PRA information is used to determine the consequence of a component rupture (increased reactor core damage probability). A procedure has also been recommended for using the resulting quantified risk estimates to determine target component rupture probability values to be maintained by inspection activities. Structural risk and reliability analysis (SRRA) calculations are then used to determine characteristics which an inspection strategy must posess in order to maintain component rupture probabilities below target values. The methodology, results of example applications, and plans for future work are discussed.

  17. Role of sphingomyelinase in mitochondrial ceramide accumulation during reperfusion.

    PubMed

    Ramírez-Camacho, I; Bautista-Pérez, R; Correa, F; Buelna-Chontal, M; Román-Anguiano, N G; Medel-Franco, M; Medina-Campos, O N; Pedraza-Chaverri, J; Cano-Martínez, A; Zazueta, C

    2016-10-01

    Ceramide accumulation in mitochondria has been associated with reperfusion damage, but the underlying mechanisms are not clearly elucidated. In this work we investigate the role of sphingomyelinases in mitochondrial ceramide accumulation, its effect on reactive oxygen species production, as well as on mitochondrial function by using the sphingomyelinase inhibitor, tricyclodecan-9-yl-xanthogenate (D609). Correlation between neutral sphingomyelinase (nSMase) activity and changes in ceramide content were performed in whole tissue and in isolated mitochondria from reperfused hearts. Overall results demonstrated that D609 treatment attenuates cardiac dysfuncion, mitochondrial injury and oxidative stress. Ceramide was accumulated in mitochondria, but not in the microsomal fraction of the ischemic-reperfused (I/R) group. In close association, the activity of nSMase increased, whereas glutathione (GSH) levels diminished in mitochondria after reperfusion. On the other hand, reduction of ceramide levels in mitochondria from I/R+D609 hearts correlated with diminished nSMase activity, coupling of oxidative phosphorylation and with mitochondrial integrity maintenance. These results suggest that mitochondrial nSMase activity contributes to compartmentation and further accumulation of ceramide in mitochondria, deregulating their function during reperfusion. PMID:27479697

  18. Localization of cholesterol in sphingomyelinase-treated fibroblasts.

    PubMed Central

    Pörn, M I; Slotte, J P

    1995-01-01

    The distribution of cellular unesterified cholesterol was studied in fibroblasts, which had been depleted of plasma membrane sphingomyelin by exposure to exogenous sphingomyelinase. This treatment has previously been shown to induce an increase in cholesterol esterification, a decrease in the biosynthesis of cholesterol, and a decreased susceptibility of cell cholesterol to oxidation with cholesterol oxidase. When the cellular localization of cholesterol was studied with fluorescent filipin staining, sphingomyelin depletion did not cause any visible changes in the filipin-cholesterol staining pattern, suggesting that the major part of cellular cholesterol was retained in the plasma membrane after sphingomyelinase treatment. After the oxidation of cell-surface cholesterol with cholesterol oxidase, the plasma membrane was no longer stained by filipin, but the plasma membrane cholesterol of sphingomyelin-depleted cells appeared to be resistant to oxidation with cholesterol oxidase when sphingomyelinase was used as an oxidation-promoting agent. However, the use of hypotonic buffer or phosphatidylcholine-specific phospholipase C together with cholesterol oxidase resulted in a complete oxidation of the cell-surface cholesterol in sphingomyelin-depleted cells, as evidenced by the filipin-cholesterol staining pattern. Similar results were obtained when [3H]cholesterol-labelled fibroblasts were used for determination of the susceptibility to cholesterol oxidation. The kinetics of [3H]cholesterol oxidation in sphingomyelin-depleted cells with cholesterol oxidase in hypotonic buffer indicated that approximately 85% of the cellular cholesterol still resided in the plasma membrane after sphingomyelin depletion. These results are contradictory to earlier reports on sphingomyelinase-induced changes in cellular cholesterol distribution and suggest that minor changes in the kinetics of cholesterol transport from the plasma membrane to the endoplasmic reticulum may be responsible

  19. Sphingomyelinase-Like Phosphodiesterase 3b Expression Levels Determine Podocyte Injury Phenotypes in Glomerular Disease

    PubMed Central

    Yoo, Tae-Hyun; Pedigo, Christopher E.; Guzman, Johanna; Correa-Medina, Mayrin; Wei, Changli; Villarreal, Rodrigo; Mitrofanova, Alla; Leclercq, Farah; Faul, Christian; Li, Jing; Kretzler, Matthias; Nelson, Robert G.; Lehto, Markku; Forsblom, Carol; Groop, Per-Henrik; Reiser, Jochen; Burke, George William

    2015-01-01

    Diabetic kidney disease (DKD) is the most common cause of ESRD in the United States. Podocyte injury is an important feature of DKD that is likely to be caused by circulating factors other than glucose. Soluble urokinase plasminogen activator receptor (suPAR) is a circulating factor found to be elevated in the serum of patients with FSGS and causes podocyte αVβ3 integrin-dependent migration in vitro. Furthermore, αVβ3 integrin activation occurs in association with decreased podocyte-specific expression of acid sphingomyelinase-like phosphodiesterase 3b (SMPDL3b) in kidney biopsy specimens from patients with FSGS. However, whether suPAR-dependent αVβ3 integrin activation occurs in diseases other than FSGS and whether there is a direct link between circulating suPAR levels and SMPDL3b expression in podocytes remain to be established. Our data indicate that serum suPAR levels are also elevated in patients with DKD. However, unlike in FSGS, SMPDL3b expression was increased in glomeruli from patients with DKD and DKD sera-treated human podocytes, where it prevented αVβ3 integrin activation by its interaction with suPAR and led to increased RhoA activity, rendering podocytes more susceptible to apoptosis. In vivo, inhibition of acid sphingomyelinase reduced proteinuria in experimental DKD but not FSGS, indicating that SMPDL3b expression levels determined the podocyte injury phenotype. These observations suggest that SMPDL3b may be an important modulator of podocyte function by shifting suPAR-mediated podocyte injury from a migratory phenotype to an apoptotic phenotype and that it represents a novel therapeutic glomerular disease target. PMID:24925721

  20. Sphingomyelinase-like phosphodiesterase 3b expression levels determine podocyte injury phenotypes in glomerular disease.

    PubMed

    Yoo, Tae-Hyun; Pedigo, Christopher E; Guzman, Johanna; Correa-Medina, Mayrin; Wei, Changli; Villarreal, Rodrigo; Mitrofanova, Alla; Leclercq, Farah; Faul, Christian; Li, Jing; Kretzler, Matthias; Nelson, Robert G; Lehto, Markku; Forsblom, Carol; Groop, Per-Henrik; Reiser, Jochen; Burke, George William; Fornoni, Alessia; Merscher, Sandra

    2015-01-01

    Diabetic kidney disease (DKD) is the most common cause of ESRD in the United States. Podocyte injury is an important feature of DKD that is likely to be caused by circulating factors other than glucose. Soluble urokinase plasminogen activator receptor (suPAR) is a circulating factor found to be elevated in the serum of patients with FSGS and causes podocyte αVβ3 integrin-dependent migration in vitro. Furthermore, αVβ3 integrin activation occurs in association with decreased podocyte-specific expression of acid sphingomyelinase-like phosphodiesterase 3b (SMPDL3b) in kidney biopsy specimens from patients with FSGS. However, whether suPAR-dependent αVβ3 integrin activation occurs in diseases other than FSGS and whether there is a direct link between circulating suPAR levels and SMPDL3b expression in podocytes remain to be established. Our data indicate that serum suPAR levels are also elevated in patients with DKD. However, unlike in FSGS, SMPDL3b expression was increased in glomeruli from patients with DKD and DKD sera-treated human podocytes, where it prevented αVβ3 integrin activation by its interaction with suPAR and led to increased RhoA activity, rendering podocytes more susceptible to apoptosis. In vivo, inhibition of acid sphingomyelinase reduced proteinuria in experimental DKD but not FSGS, indicating that SMPDL3b expression levels determined the podocyte injury phenotype. These observations suggest that SMPDL3b may be an important modulator of podocyte function by shifting suPAR-mediated podocyte injury from a migratory phenotype to an apoptotic phenotype and that it represents a novel therapeutic glomerular disease target. PMID:24925721

  1. ASM Conference on Prokaryotic Development

    SciTech Connect

    Kaplan, H. B.

    2005-07-13

    Support was provided by DOE for the 2nd ASM Conference on Prokaryotic Development. The final conference program and abstracts book is attached. The conference presentations are organized around topics that are central to the current research areas in prokaryotic development. The program starts with topics that involve relatively simple models systems and ends with systems that are more complex. The topics are: i) the cell cycle, ii) the cytoskeleton, iii) morphogenesis, iv) developmental transcription, v) signaling, vi) multicellularity, and vii) developmental diversity and symbiosis. The best-studied prokaryotic development model systems will be highlighted at the conference through research presentations by leaders in the field. Many of these systems are also model systems of relevance to the DOE mission including carbon sequestration (Bradyrizobium, Synechococcus), energy production (Anabaena, Rhodobacter) and bioremediation (Caulobacter, Mesorhizobium). In addition, many of the highlighted organisms have important practical applications; the actinomycetes and myxobacteria produce antimicrobials that are of commercial interest. It is certain that the cutting-edge science presented at the conference will be applicable to the large group of bacteria relevant to the DOE mission.

  2. Sphingosine inhibits sphingomyelinase-induced cholesteryl ester formation in cultured fibroblasts.

    PubMed

    Härmälä, A S; Pörn, M I; Slotte, J P

    1993-12-01

    We have in this study examined the effects of sphingosine, a possible secondary degradation product following sphingomyelin hydrolysis, on cholesterol homeostasis in cultured human fibroblasts treated with sphingomyelinase. The activation of cholesterol esterification, which resulted from the degradation of plasma membrane sphingomyelin (by sphingomyelinase), was observed to be effectively blocked by sphingosine (half-maximal dose 6-7 microM). The inhibitory action of sphingosine could not be reproduced with other amines (e.g., dodecyl amine or imipramine). The onset of inhibition of cholesteryl ester formation by sphingosine was rapid (maximal effect within 15 min). Sphingosine itself had no spontaneous effects on the distribution of cellular cholesterol. At concentrations below 10 microM, sphingosine was not cytotoxic, as determined by cellular trypan blue permeability. The inhibitory action of sphingosine on cholesteryl ester formation apparently did not result from a direct inhibition of acyl-CoA cholesterol acyltransferase (ACAT), since the activity of this enzyme was unaffected by sphingosine (10 microM) in a cell-free homogenate, using [14C]oleoyl-CoA as a co-substrate. Sphingosine was also unable to prevent the formation of activated fatty acids (oleoyl-CoA), since acyl-CoA synthetase activity in a cell-free homogenate was not inhibited by sphingosine (at 5 microM). The cellular cholesteryl ester cycle (i.e., the neutral cholesteryl ester hydrolase) was unaffected by sphingosine (at 5 microM). Down-regulation of PKC activity (24 h exposure of cells to 100 nM (62 ng/ml) phorbol ester) did not affect the sphingomyelinase-induced stimulation of [3H]cholesteryl ester formation. In addition, the sphingosine-induced inhibition of [3H]cholesteryl ester formation was not reversed in the presence of phorbol ester (short-term exposures), suggesting that the effect of sphingosine was not mediated via PKC. In conclusion, we have shown that sphingosine is an inhibitor

  3. The ASME Code today -- Challenges, threats, opportunities

    SciTech Connect

    Canonico, D.A.

    1995-12-01

    Since its modest beginning as a single volume in 1914 the ASME Code, or some of its parts, is recognized today in 48 of the United States and all providence`s of Canada. The ASME Code today is composed of 25 books including two Code Case books. These books cover the new construction of boilers and pressure vessels and the new construction and In-Service-Inspection of Nuclear Power Plant components. The ASME accredits all manufacturers of boilers and pressure vessels built to the ASME Code. There are approximately 7650 symbol stamps issued throughout the world. Over 23% of the symbol stamps have been issued outside the USA and Canada. The challenge to the ASME Code is to be accepted as the world standard for pressure boundary components. There are activities underway to achieve that goal. The ASME Code is being revised to make it a more friendly document to entities outside of North America. To achieve that end there are specific tasks underway which are described here.

  4. Sphingomyelinase D/Ceramide 1-Phosphate in Cell Survival and Inflammation

    PubMed Central

    Rivera, Io-Guané; Ordoñez, Marta; Presa, Natalia; Gomez-Larrauri, Ana; Simón, Jorge; Trueba, Miguel; Gomez-Muñoz, Antonio

    2015-01-01

    Sphingolipids are major constituents of biological membranes of eukaryotic cells. Many studies have shown that sphingomyelin (SM) is a major phospholipid in cell bilayers and is mainly localized to the plasma membrane of cells, where it serves both as a building block for cell architecture and as a precursor of bioactive sphingolipids. In particular, upregulation of (C-type) sphingomyelinases will produce ceramide, which regulates many physiological functions including apoptosis, senescence, or cell differentiation. Interestingly, the venom of some arthropodes including spiders of the genus Loxosceles, or the toxins of some bacteria such as Corynebacterium tuberculosis, or Vibrio damsela possess high levels of D-type sphingomyelinase (SMase D). This enzyme catalyzes the hydrolysis of SM to yield ceramide 1-phosphate (C1P), which promotes cell growth and survival and is a potent pro-inflammatory agent in different cell types. In particular, C1P stimulates cytosolic phospholipase A2 leading to arachidonic acid release and the subsequent formation of eicosanoids, actions that are all associated to the promotion of inflammation. In addition, C1P potently stimulates macrophage migration, which has also been associated to inflammatory responses. Interestingly, this action required the interaction of C1P with a specific plasma membrane receptor, whereas accumulation of intracellular C1P failed to stimulate chemotaxis. The C1P receptor is coupled to Gi proteins and activates of the PI3K/Akt and MEK/ERK1-2 pathways upon ligation with C1P. The proposed review will address novel aspects on the control of inflammatory responses by C1P and will highlight the molecular mechanisms whereby C1P exerts these actions. PMID:25938271

  5. Neutral sphingomyelinase 2: a novel target in cigarette smoke-induced apoptosis and lung injury.

    PubMed

    Filosto, Simone; Castillo, Sianna; Danielson, Aaron; Franzi, Lisa; Khan, Elaine; Kenyon, Nick; Last, Jerold; Pinkerton, Kent; Tuder, Rubin; Goldkorn, Tzipora

    2011-03-01

    Chronic obstructive pulmonary disease (COPD) is caused by exposure to cigarette smoke (CS). One mechanism of CS-induced lung injury is aberrant generation of ceramide, which leads to elevated apoptosis of epithelial and endothelial cells in the alveolar spaces. Recently, we discovered that CS-induced ceramide generation and apoptosis in pulmonary cells is governed by neutral sphingomyelinase (nSMase) 2. In the current experiments, we expanded our studies to investigate whether nSMase2 governs ceramide generation and apoptosis in vivo using rodent and human models of CS-induced lung injury. We found that exposure of mice or rats to CS leads to colocalizing elevations of ceramide levels and terminal deoxynucleotidyl transferase mediated X-dUTP nick end labeling-positive cells in lung tissues. These increases are nSMase2 dependent, and are abrogated by treatment with N-acetyl cysteine or anti-nSMase2 small interfering RNA (siRNA). We further showed that mice that are heterozygous for nSMase2 demonstrate significant decrease in ceramide generation after CS exposure, whereas acidic sphingomyelinase (aSMase) knockout mice maintain wild-type ceramide levels, confirming our previous findings (in human airway epithelial cells) that only nSMase2, and not aSMase, is activated by CS exposure. Lastly, we found that lung tissues from patients with emphysema (smokers) display significantly higher levels of nSMase2 expression compared with lung tissues from healthy control subjects. Taken together, these data establish the central in vivo role of nSMase2 in ceramide generation, aberrant apoptosis, and lung injury under CS exposure, underscoring its promise as a novel target for the prevention of CS-induced airspace destruction. PMID:20448054

  6. ASME Material Challenges for Advanced Reactor Concepts

    SciTech Connect

    Piyush Sabharwall; Ali Siahpush

    2013-07-01

    This study presents the material Challenges associated with Advanced Reactor Concept (ARC) such as the Advanced High Temperature Reactor (AHTR). ACR are the next generation concepts focusing on power production and providing thermal energy for industrial applications. The efficient transfer of energy for industrial applications depends on the ability to incorporate cost-effective heat exchangers between the nuclear heat transport system and industrial process heat transport system. The heat exchanger required for AHTR is subjected to a unique set of conditions that bring with them several design challenges not encountered in standard heat exchangers. The corrosive molten salts, especially at higher temperatures, require materials throughout the system to avoid corrosion, and adverse high-temperature effects such as creep. Given the very high steam generator pressure of the supercritical steam cycle, it is anticipated that water tube and molten salt shell steam generators heat exchanger will be used. In this paper, the ASME Section III and the American Society of Mechanical Engineers (ASME) Section VIII requirements (acceptance criteria) are discussed. Also, the ASME material acceptance criteria (ASME Section II, Part D) for high temperature environment are presented. Finally, lack of ASME acceptance criteria for thermal design and analysis are discussed.

  7. Sphingomyelinase Activity of Trichomonas vaginalis Extract and Subfractions

    PubMed Central

    González-Salazar, Francisco; Garza-González, Jesús N.; Hernandez-Luna, Carlos E.; Mata-Cárdenas, Benito David; Carranza-Rosales, Pilar; Castro-Garza, Jorge Enrique; Hernández-García, Magda Elizabeth; Vargas-Villarreal, Javier

    2013-01-01

    Trichomoniasis is one of the most common acute sexually transmitted curable diseases, and it is disseminated worldwide generating more than 170 million cases annually. Trichomonas vaginalis is the parasite that causes trichomoniasis and has the ability to destroy cell monolayers of the vaginal mucosa in vitro. Sphingomyelinases (SMase) are enzymes that catalyze the hydrolysis of sphingomyelin into ceramide and phosphorylcholine. Ceramide appears to be a second messenger lipid in programmed apoptosis, cell differentiation, and cell proliferation. Sphingomyelinase is probably a major source of ceramide in cells. Signal transduction mediated by ceramide leads cells to produce cytokine induced apoptosis during several inflammatory responses. SMase are also relevant toxins in several microorganisms. The main objective of this research is to identify SMase activity of T. vaginalis in the total extract (TE), P30, and S30 subfractions from brooked trophozoites. It was found that these fractions of T. vaginalis have SMase activity, which comes principally from P30 subfraction and was mainly type C. Enzymatic activity of SMase increased linearly with time and is pH dependent with two peaks by pH 5.5 and pH 7.5. The addition of manganese to the reaction mixture increased the SMase activity by 1.97. PMID:24024206

  8. Improvement of ASME NH for Grade 91

    SciTech Connect

    Bernard Riou

    2007-10-09

    This report has been prepared in the context of Task 3 of the ASME/DOE Gen IV material project. It has been identified that creep-fatigue evaluation procedures presently available in ASME (1) and RCC-MR (2) have been mainly developed for austenitic stainless steels and may not be suitable for cyclic softening materials such as mod 9 Cr 1 Mo steel (grade 91). The aim of this document is, starting from experimental test results, to perform a review of the procedures and, if necessary, provide recommendations for their improvements.

  9. Proceedings: 1985 ASME-EPRI Radwaste Workshop

    SciTech Connect

    1985-07-01

    This year's ASME-EPRI workshop targeted three main topics: meeting federal requirements, using mobile services, and cutting costs. A highlight of the workshop was a detailed report on the mobile services for radwaste management that are now available to utilities from outside contractors.

  10. Identification of New Sphingomyelinases D in Pathogenic Fungi and Other Pathogenic Organisms

    PubMed Central

    Neshich, Goran; Ortega, José Miguel; Granier, Claude; Chávez-Olortegui, Carlos; Molina, Franck; Felicori, Liza

    2013-01-01

    Sphingomyelinases D (SMases D) or dermonecrotic toxins are well characterized in Loxosceles spider venoms and have been described in some strains of pathogenic microorganisms, such as Corynebacterium sp. After spider bites, the SMase D molecules cause skin necrosis and occasional severe systemic manifestations, such as acute renal failure. In this paper, we identified new SMase D amino acid sequences from various organisms belonging to 24 distinct genera, of which, 19 are new. These SMases D share a conserved active site and a C-terminal motif. We suggest that the C-terminal tail is responsible for stabilizing the entire internal structure of the SMase D Tim barrel and that it can be considered an SMase D hallmark in combination with the amino acid residues from the active site. Most of these enzyme sequences were discovered from fungi and the SMase D activity was experimentally confirmed in the fungus Aspergillus flavus. Because most of these novel SMases D are from organisms that are endowed with pathogenic properties similar to those evoked by these enzymes alone, they might be associated with their pathogenic mechanisms. PMID:24223912

  11. ASME Task Group report on primary stress

    SciTech Connect

    Pastor, T.P.; Hechmer, J.

    1997-02-01

    This paper considers the subject of primary stress as applied in pressure vessel design carried out in accordance with the ASME Boiler and Pressure Vessel Code (1992). The paper is the result of discussions held by members of the Task Group on Primary Stress. Specific subjects discussed include methods for calculating primary stresses, the ASME limits on primary stresses and their meaning, use of the Code stress classification table in pressure vessel design, and commentary on the use of state-of-the-art analysis techniques to design pressure vessels and satisfy Code primary stress limits. A modified definition for primary stress is given, and examples for evaluating primary stresses for different geometries is provided.

  12. Current Activities of the ASME Subgroup NUPACK

    SciTech Connect

    Gerald M. Foster; D. Keith Morton; Paul McConnell

    2007-10-01

    Current activities of the American Society of Mechanical Engineers (ASME), Section III Subgroup on Containment Systems for Spent Fuel High-Level Waste Transport Packagings (also known as Subgroup NUPACK) are reviewed with emphasis on the recent revision of Subsection WB. Also, brief insightson new proposals for the development of rules for internal support structures and for a strain-based acceptance criteria are provided.

  13. Proteolysis sensitizes LDL particles to phospholipolysis by secretory phospholipase A2 group V and secretory sphingomyelinase

    PubMed Central

    Plihtari, Riia; Hurt-Camejo, Eva; Öörni, Katariina; Kovanen, Petri T.

    2010-01-01

    LDL particles that enter the arterial intima become exposed to proteolytic and lipolytic modifications. The extracellular hydrolases potentially involved in LDL modification include proteolytic enzymes, such as chymase, cathepsin S, and plasmin, and phospholipolytic enzymes, such as secretory phospholipases A2 (sPLA2-IIa and sPLA2-V) and secretory acid sphingomyelinase (sSMase). Here, LDL was first proteolyzed and then subjected to lipolysis, after which the effects of combined proteolysis and lipolysis on LDL fusion and on binding to human aortic proteoglycans (PG) were studied. Chymase and cathepsin S led to more extensive proteolysis and release of peptide fragments from LDL than did plasmin. sPLA2-IIa was not able to hydrolyze unmodified LDL, and even preproteolysis of LDL particles failed to enhance lipolysis by this enzyme. However, preproteolysis with chymase and cathepsin S accelerated lipolysis by sPLA2-V and sSMase, which resulted in enhanced fusion and proteoglycan binding of the preproteolyzed LDL particles. Taken together, the results revealed that proteolysis sensitizes the LDL particles to hydrolysis by sPLA2-V and sSMase. By promoting fusion and binding of LDL to human aortic proteoglycans, the combination of proteolysis and phospholipolysis of LDL particles potentially enhances extracellular accumulation of LDL-derived lipids during atherogenesis. PMID:20124257

  14. The neutral sphingomyelinase-2 is involved in angiogenic signaling triggered by oxidized LDL.

    PubMed

    Camaré, Caroline; Augé, Nathalie; Pucelle, Mélanie; Saint-Lebes, Bertrand; Grazide, Marie-Hélène; Nègre-Salvayre, Anne; Salvayre, Robert

    2016-04-01

    Capillaries of the external part of the normal arterial wall constitute the vasa vasorum network. In atherosclerotic lesions, neovascularization occurs in areas of intimal hyperplasia where it may promote plaque expansion, and intraplaque hemorrhage. Oxidized LDL that are present in atherosclerotic areas activate various angiogenic signaling pathways, including reactive oxygen species and the sphingosine kinase/sphingosine-1-phosphate pathway. We aimed to investigate whether oxidized LDL-induced angiogenesis requires neutral sphingomyelinase-2 activation and the neutral sphingomyelinase-2/sphingosine kinase-1 pathway. The role of neutral sphingomyelinase-2 in angiogenic signaling was investigated in Human Microvascular Endothelial Cells (HMEC-1) forming capillary tube on Matrigel and in vivo in the Matrigel plug assay in C57BL/6 mice and in the chicken chorioallantoic membrane model. Low concentration of human oxidized LDL elicits HMEC-1 capillary tube formation and neutral sphingomyelinase-2 activation, which were blocked by neutral sphingomyelinase-2 inhibitors, GW4869 and specific siRNA. This angiogenic effect was mimicked by low concentration of C6-Ceramide and was inhibited by sphingosine kinase-1 inhibitors. Upstream of neutral sphingomyelinase-2, oxidized LDL-induced activation required LOX-1, reactive oxygen species generation by NADPH oxidase and p38-MAPK activation. Inhibition of sphingosine kinase-1 blocked the angiogenic response and triggered HMEC-1 apoptosis. Low concentration of oxidized LDL was angiogenic in vivo, both in the Matrigel plug assay in mice and in the chorioallantoic membrane model, and was blocked by GW4869. In conclusion, low oxLDL concentration triggers sprouting angiogenesis that involves ROS-induced activation of the neutral sphingomyelinase-2/sphingosine kinase-1 pathway, and is effectively inhibited by GW4869. PMID:26855418

  15. Niemann-Pick disease: prenatal diagnoses and studies of sphingomyelinase activities.

    PubMed

    Wenger, D A; Wharton, C; Sattler, M; Clark, C

    1978-01-01

    Five pregnancies were monitored for couples at-risk for having a child with some form of Niemann-Pick disease (NPD). Three of these were for the classic Type A form in which affected children usually have less than 1% of normal sphingomyelinase activity. Two of these pregnancies were terminated after the cultured amniotic-fluid cells were determined to have less than 1% of normal sphingomyelinase activity (0.4 and 0.6 nmole/mg protein/hr versus the control mean of 61.7). In the other pregnancy at risk for Type A NPD near normal activity was measured and it was continued to term. The two other pregnancies were monitored for couples in which severely affected children were found to have partially deficient sphingomyelinase activity (about 20% of normal) in cultured skin fibroblasts. Cultural amniotic-fluid cells from one of these pregnancies also had about 20% of control sphingomyelinase activity, but the woman underwent a spontaneous abortion soon after the cells were received and no studies on the fetus were done. The other sample was taken at the time of abortion for social reasons. In this case the cultured amniotic-fluid cells and cultured fetal skin fibroblasts gave normal sphingomyelinase activity. Enzymatic studies on tissues from the two fetuses predicted to be affected with Type A NPD confirmed the prenatal diagnosis. Studies of sphingomyelinase activity in the brains from these fetuses and from a child who died with Type A NPD indicated significant levels of activity when measured at pH 7.4 in the presence of magnesium. The higher level of the pH 7.4 sphingomyelinase activity in developing brain may indicate some important role in normal brain development. PMID:233699

  16. Inhibitors of the sphingomyelin cycle: Sphingomyelin synthases and sphingomyelinases.

    PubMed

    Adada, Mohamad; Luberto, Chiara; Canals, Daniel

    2016-05-01

    Sphingolipids are a class of bioactive lipids, which are key modulators of an increasing number of physiologic and pathophysiologic processes that include cell cycle, apoptosis, angiogenesis, stress and inflammatory responses. Sphingomyelin is an important structural component of biological membranes, and one of the end-points in the synthesis of sphingolipids. Mainly synthetized in the Golgi apparatus, sphingomyelin is transported to all other biological membranes. Upon stimulation, sphingomyelin can be hydrolyzed to ceramide by 5 different sphingomyelinases. The diversity and cellular topology of ceramide allow it to exert multiple biologies. Furthermore, ceramide can be metabolized to many other bioactive sphingolipids. Ceramide, coming from sphingomyelin or other complex sphingolipids, can be hydrolyzed to sphingosine, which can easily change cellular localization. In turn, sphingosine can be recycled to ceramide and to sphingomyelin in the endoplasmic reticulum, completing the sphingomyelin cycle. Our understanding of the roles of various sphingolipids in the regulation of different cellular processes has come from studying the enzymes that regulate these sphingolipids, and their manipulation. The use of pharmacologic inhibitors has been critical for their study, as well as being promising bullets for disease treatment. Some of these diseases involving the sphingomyelin cycle include cancer, inflammation, atherosclerosis, diabetes and some rare diseases such as Niemann-Pick disease. This review will focus on the enzymes involved in the sphingomyelin cycle, their history, and their involvement in pathophysiological processes. Finally, it will describe in details all the small molecules that are being used to inhibit these enzymes and their use in therapeutics. PMID:26200918

  17. ASME post construction pressure technology codes

    SciTech Connect

    Sims, J.R.

    1996-12-01

    The need to continue to operate pressurized equipment and other facilities in a safe, reliable and cost effective manner has led to the development of many new approaches to in-service inspection, flaw evaluation, and repair. Interest on the part of users, regulatory authorities and others in standardizing these approaches has led to the formation of a new ASME Main Committee on Post Construction under the Board on Pressure Technology Codes and Standards, and a new Division of the Pressure Vessel Research Council on Continued Operation of Equipment. This paper provides a brief overview of these activities.

  18. First results of a scientific validation of Swarm ASM data

    NASA Astrophysics Data System (ADS)

    Chulliat, Arnaud; Alken, Patrick; Hulot, Gauthier

    2014-05-01

    The Absolute Scalar Magnetometer (ASM) is an optically pumped helium-4 magnetometer providing scalar measurements of the Earth's magnetic field onboard each of the three Swarm satellites. Its performance (accuracy, noise level, data cleanliness) is expected to be unprecedented among satellite scalar magnetometers. The main function of the ASMs during the Swarm mission will be to calibrate the vector field magnetometers. As part of the commissioning of the ASM instrument, we will compare preliminary ASM data with predictions from various recent geomagnetic models. Special focus will be placed on the DMSP-MAG-1 model, which relies on geomagnetic data from the Defense Meteorological Satellite Program (DMSP) until 2013.

  19. Identification of Novel Anionic Phospholipid Binding Domains in Neutral Sphingomyelinase 2 with Selective Binding Preference*

    PubMed Central

    Wu, Bill X.; Clarke, Christopher J.; Matmati, Nabil; Montefusco, David; Bartke, Nana; Hannun, Yusuf A.

    2011-01-01

    Sphingolipids such as ceramide are recognized as vital regulators of many biological processes. Neutral sphingomyelinase 2 (nSMase2) is one of the key enzymes regulating ceramide production. It was previously shown that the enzymatic activity of nSMase2 was dependent on anionic phospholipids (APLs). In this study, the structural requirements for APL-selective binding of nSMase2 were determined and characterized. Using lipid-protein overlay assays, nSMase2 interacted specifically and directly with several APLs, including phosphatidylserine and phosphatidic acid. Lipid-protein binding studies of deletion mutants identified two discrete APL binding domains in the N terminus of nSMase2. Further, mutagenesis experiments pinpointed the core sequences and major cationic amino acids in the domains that are necessary for the cooperative activation of nSMase2 by APLs. The first domain included the first amino-terminal hydrophobic segment and Arg-33, which were essential for nSMase2 to interact with APLs. The second binding domain was comprised of the second hydrophobic segment and Arg-92 and Arg-93. Moreover, mutation of one or both domains decreased APL binding and APL-dependent catalytic activity of nSMase2. Further, mutation of both domains in nSMase2 reduced its plasma membrane localization. Finally, these binding domains are also important for the capability of nSMase2 to rescue the defects of yeast lacking the nSMase homologue, ISC1. In conclusion, these data have identified the APL binding domains of nSMase2 for the first time. The analysis of interactions between nSMase2 and APLs will contribute to our understanding of signaling pathways mediated by sphingolipid metabolites. PMID:21550973

  20. Role of sph2 Gene Regulation in Hemolytic and Sphingomyelinase Activities Produced by Leptospira interrogans

    PubMed Central

    Narayanavari, Suneel A.; Lourdault, Kristel; Sritharan, Manjula; Haake, David A.; Matsunaga, James

    2015-01-01

    Pathogenic members of the genus Leptospira are the causative agents of leptospirosis, a neglected disease of public and veterinary health concern. Leptospirosis is a systemic disease that in its severest forms leads to renal insufficiency, hepatic dysfunction, and pulmonary failure. Many strains of Leptospira produce hemolytic and sphingomyelinase activities, and a number of candidate leptospiral hemolysins have been identified based on sequence similarity to well-characterized bacterial hemolysins. Five of the putative hemolysins are sphingomyelinase paralogs. Although recombinant forms of the sphingomyelinase Sph2 and other hemolysins lyse erythrocytes, none have been demonstrated to contribute to the hemolytic activity secreted by leptospiral cells. In this study, we examined the regulation of sph2 and its relationship to hemolytic and sphingomyelinase activities produced by several L. interrogans strains cultivated under the osmotic conditions found in the mammalian host. The sph2 gene was poorly expressed when the Fiocruz L1-130 (serovar Copenhageni), 56601 (sv. Lai), and L495 (sv. Manilae) strains were cultivated in the standard culture medium EMJH. Raising EMJH osmolarity to physiological levels with sodium chloride enhanced Sph2 production in all three strains. In addition, the Pomona subtype kennewicki strain LC82-25 produced substantially greater amounts of Sph2 during standard EMJH growth than the other strains, and sph2 expression increased further by addition of salt. When 10% rat serum was present in EMJH along with the sodium chloride supplement, Sph2 production increased further in all strains. Osmotic regulation and differences in basal Sph2 production in the Manilae L495 and Pomona strains correlated with the levels of secreted hemolysin and sphingomyelinase activities. Finally, a transposon insertion in sph2 dramatically reduced hemolytic and sphingomyelinase activities during incubation of L. interrogans at physiologic osmolarity

  1. Enhanced colonic tumorigenesis in alkaline sphingomyelinase (NPP7) knockout mice.

    PubMed

    Chen, Ying; Zhang, Ping; Xu, Shu-Chang; Yang, Liping; Voss, Ulrikke; Ekblad, Eva; Wu, Yunjin; Min, Yalan; Hertervig, Erik; Nilsson, Åke; Duan, Rui-Dong

    2015-01-01

    Intestinal alkaline sphingomyelinase (alk-SMase) generates ceramide and inactivates platelet-activating factor (PAF) and was previously suggested to have anticancer properties. The direct evidence is still lacking. We studied colonic tumorigenesis in alk-SMase knockout (KO) mice. Formation of aberrant crypt foci (ACF) was examined after azoxymethane (AOM) injection. Tumor was induced by AOM alone, a conventional AOM/dextran sulfate sodium (DSS) treatment, and an enhanced AOM/DSS method. β-Catenin was determined by immunohistochemistry, PAF levels by ELISA, and sphingomyelin metabolites by mass spectrometry. Without treatment, spontaneous tumorigenesis was not identified but the intestinal mucosa appeared thicker in KO than in wild-type (WT) littermates. AOM alone induced more ACF in KO mice but no tumors 28 weeks after injection. However, combination of AOM/DSS treatments induced colonic tumors and the incidence was significantly higher in KO than in WT mice. By the enhanced AOM/DSS method, tumor number per mouse increased 4.5 times and tumor size 1.8 times in KO compared with WT mice. Although all tumors were adenomas in WT mice, 32% were adenocarcinomas in KO mice. Compared with WT mice, cytosol expression of β-catenin was significantly increased and nuclear translocation in tumors was more pronounced in KO mice. Lipid analysis showed decreased ceramide in small intestine and increased sphingosine-1-phosphate (S1P) in both small intestine and colon in nontreated KO mice. PAF levels in feces were significantly higher in the KO mice after AOM/DSS treatment. In conclusion, lack of alk-SMase markedly increases AOM/DSS-induced colonic tumorigenesis associated with decreased ceramide and increased S1P and PAF levels. PMID:25381265

  2. 78 FR 37848 - ASME Code Cases Not Approved for Use

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-06-24

    ...The U.S. Nuclear Regulatory Commission (NRC) is issuing for public comment draft regulatory guide (DG), DG-1233, ``ASME Code Cases not Approved for Use.'' This regulatory guide lists the American Society of Mechanical Engineers (ASME) Code Cases that the NRC has determined not to be acceptable for use on a generic...

  3. Kinetic steps for the hydrolysis of sphingomyelin by Bacillus cereus sphingomyelinase in lipid monolayers.

    PubMed

    Fanani, M L; Maggio, B

    2000-11-01

    The sphingomyelinase (Sphmase) activity degrading sphingomyelin (Sphm) monolayers shows a slow-reaction latency period before exhibiting constant rate catalysis. These two kinetic regions are regulated independently by the lateral surface pressure and by lipids that are biomodulators of cell function such as ceramide, glycosphingolipids, fatty acids, and lysophospholipids. Knowledge of the interfacial adsorption of Sphmase, precatalytic activation, initiation of effective catalysis, and the corresponding kinetic parameters is necessary for studying the level at which different lipids modulate the activity. We dissected some kinetic steps and determined the rate constants for degradation of Sphm, under controlled intermolecular organization, by Sphmase. Six models, adapted to two dimensions, were used to elucidate possible mechanisms for the interfacial activation of Sphmase during the lag time. The models consider enzyme binding to the substrate monolayer and a subsequent, essentially irreversible interfacial activation; this is supported experimentally by monolayer transfer experiments. Some mechanisms involve enzyme-substrate binding and associated states of the enzyme in the bulk subphase or at the interface, prior to complete activation. The activity of Sphmase is consistent with kinetics involving enzyme partitioning into the interface followed by substrate association, and by a process that proceeds with bimolecular kinetic dependence on the interfacial Sphmase concentration, and a subsequent slow step of activation. A possible equilibrium between the apparent monomolecular and bimolecular activated states of the interfacial enzyme, coupled to a slow activation, constitute rate-limiting steps that can explain the existence of lag time and the achievement of a maximum constant rate of degradation of Sphm monolayers by Sphmase. PMID:11060353

  4. Sphingomyelinase D from venoms of Loxosceles spiders: evolutionary insights from cDNA sequences and gene structure.

    PubMed

    Binford, Greta J; Cordes, Matthew H J; Wells, Michael A

    2005-04-01

    Loxosceles spider venoms cause dermonecrosis in mammalian tissues. The toxin sphingomyelinase D (SMaseD) is a sufficient causative agent in lesion formation and is only known in these spiders and a few pathogenic bacteria. Similarities between spider and bacterial SMaseD in molecular weights, pIs and N-terminal amino acid sequence suggest an evolutionary relationship between these molecules. We report three cDNA sequences from venom-expressed mRNAs, analyses of amino acid sequences, and partial characterization of gene structure of SMaseD homologs from Loxosceles arizonica with the goal of better understanding the evolution of this toxin. Sequence analyses indicate SMaseD is a single domain protein and a divergent member of the ubitiquous, broadly conserved glycerophosphoryl diester phosphodiesterase family (GDPD). Bacterial SMaseDs are not identifiable as homologs of spider SMaseD or GDPD family members. Amino acid sequence similarities do not afford clear distinction between independent origin of toxic SMaseD activity in spiders and bacteria and origin in one lineage by ancient horizontal transfer from the other. The SMaseD genes span at least 6500bp and contain at least 5 introns. Together, these data indicate L. arizonica SMaseD has been evolving within a eukaryotic genome for a long time ruling out origin by recent transfer from bacteria. PMID:15777950

  5. Neutral sphingomyelinase-2 is a redox sensitive enzyme: role of catalytic cysteine residues in regulation of enzymatic activity through changes in oligomeric state

    PubMed Central

    Dotson, P. Patrick; Karakashian, Alexander A.; Nikolova-Karakashian, Mariana N.

    2015-01-01

    Neutral sphingomyelinase-2 (nSMase-2) is the major sphingomyelinase activated in response to pro-inflammatory cytokines and during oxidative stress. It is a membrane-bound 655 amino acid protein containing 22 cysteine residues. In this study, we expressed recombinant mouse nSMase-2 protein in Escherichia coli, and investigated whether nSMase-2 is a redox sensitive enzyme. Our results demonstrate that nSMase-2 exists as both monomers and multimers that are associated with high and low enzymatic activity respectively. Mutational analysis of nSMase-2 identified within its C-terminal catalytic domain several oxidant-sensitive cysteine residues that were shown to be involved in enzyme oligomerization. Changing Cys617 to Ser for example is a gain-of-function mutation associated with a decreased propensity for oligomerization. Alternatively, nSMase-2 expression in a bacterial strain that lacks endogenous thioredoxin, Rosetta-gami2, results in increased oligomer formation and lower enzyme activity. Phenotypic rescue was accomplished by treating nSMase-2 lysates with recombinant human thioredoxin. This indicates that nSMase-2 may be a novel substrate for thioredoxin. FRET analysis confirmed the presence of nSMase-2 multimers in mammalian HEK cells and their localization to the plasma membrane. In conclusion, our results identify nSMase-2 as a redox-sensitive enzyme, whose basal activity is influenced by thioredoxin-mediated changes in its oligomeric state. PMID:25287744

  6. Neutral sphingomyelinase-2 is a redox sensitive enzyme: role of catalytic cysteine residues in regulation of enzymatic activity through changes in oligomeric state.

    PubMed

    Dotson, P Patrick; Karakashian, Alexander A; Nikolova-Karakashian, Mariana N

    2015-02-01

    Neutral sphingomyelinase-2 (nSMase-2) is the major sphingomyelinase activated in response to pro-inflammatory cytokines and during oxidative stress. It is a membrane-bound 655 amino acid protein containing 22 cysteine residues. In this study, we expressed recombinant mouse nSMase-2 protein in Escherichia coli, and investigated whether nSMase-2 is a redox sensitive enzyme. Our results demonstrate that nSMase-2 exists as both monomers and multimers that are associated with high and low enzymatic activity respectively. Mutational analysis of nSMase-2 identified within its C-terminal catalytic domain several oxidant-sensitive cysteine residues that were shown to be involved in enzyme oligomerization. Changing Cys(617) to Ser for example is a gain-of-function mutation associated with a decreased propensity for oligomerization. Alternatively, nSMase-2 expression in a bacterial strain that lacks endogenous thioredoxin, Rosetta-gami2, results in increased oligomer formation and lower enzyme activity. Phenotypic rescue was accomplished by treating nSMase-2 lysates with recombinant human thioredoxin. This indicates that nSMase-2 may be a novel substrate for thioredoxin. FRET analysis confirmed the presence of nSMase-2 multimers in mammalian HEK cells and their localization to the plasma membrane. In conclusion, our results identify nSMase-2 as a redox-sensitive enzyme, whose basal activity is influenced by thioredoxin-mediated changes in its oligomeric state. PMID:25287744

  7. In vitro effects of fat, FA, and cholesterol on sphingomyelin hydrolysis induced by rat intestinal alkaline sphingomyelinase.

    PubMed

    Liu, Jian-Jun; Nilsson, Ake; Duan, Rui-Dong

    2002-05-01

    Dietary sphingomyelin (SM) may have regulatory effects on cell proliferation and tumorigenesis in the colon. Alkaline sphingomyelinase (SMase) is the major enzyme responsible for hydrolysis of SM in the gut. Previously we purified the enzyme and showed that the presence of glycerophospholipids inhibited SM hydrolysis induced by alkaline SMase in vitro. In the present work, we studied the effects of TG, DG, FA, ceramide, and cholesterol on SM hydrolysis catalyzed by purified alkaline SMase. The results showed that both TG (triolein and tristearin) and DG (1,2-dioleoyl-sn-glycerol and 1,2-distearoyl-rac-glycerol) inhibited the activity of alkaline SMase. 1-Monooleoyl-rac-glycerol, 1-monostearoyl-rac-glycerol, stearic acid, oleic acid, linoleic acid, linolenic acid, and arachidonic acid stimulated the activity of alkaline SMase at 0.4-0.8 mM concentrations but inhibited the enzyme at higher concentrations. There was no difference between the effects induced by saturated and unsaturated FA. A short-chain FA such as lauric acid had a stronger stimulatory effect at low concentrations and weaker inhibitory effect at high concentrations than long-chain FA. Choosing linoleic acid as an example, we found that FA had similar effects on both alkaline SMase and neutral SMase. Cholesterol and ceramide when mixed with FA to increase its solubility in bile salt micelles inhibited SMase activity. In conclusion, glycerides, FA, ceramide, and cholesterol influence SM hydrolysis catalyzed by intestinal alkaline SMase. The presence of lipids in the diet may thus influence the course of SM digestion in the gut and thereby the exposure of colon to SM metabolites. PMID:12056588

  8. Accelerator System Model (ASM) user manual with physics and engineering model documentation. ASM version 1.0

    SciTech Connect

    1993-07-01

    The Accelerator System Model (ASM) is a computer program developed to model proton radiofrequency accelerators and to carry out system level trade studies. The ASM FORTRAN subroutines are incorporated into an intuitive graphical user interface which provides for the {open_quotes}construction{close_quotes} of the accelerator in a window on the computer screen. The interface is based on the Shell for Particle Accelerator Related Codes (SPARC) software technology written for the Macintosh operating system in the C programming language. This User Manual describes the operation and use of the ASM application within the SPARC interface. The Appendix provides a detailed description of the physics and engineering models used in ASM. ASM Version 1.0 is joint project of G. H. Gillespie Associates, Inc. and the Accelerator Technology (AT) Division of the Los Alamos National Laboratory. Neither the ASM Version 1.0 software nor this ASM Documentation may be reproduced without the expressed written consent of both the Los Alamos National Laboratory and G. H. Gillespie Associates, Inc.

  9. Neutral sphingomyelinase 2 modulates cytotoxic effects of protopanaxadiol on different human cancer cells

    PubMed Central

    2013-01-01

    Background Some of ginsenosides, root extracts from Panax ginseng, exert cytotoxicity against cancer cells through disruption of membrane subdomains called lipid rafts. Protopanaxadiol (PPD) exhibits the highest cytotoxic effect among 8 ginsenosides which we evaluated for anti-cancer activity. We investigated if PPD disrupts lipid rafts in its cytotoxic effects and also the possible mechanisms. Methods Eight ginsenosides were evaluated using different cancer cells and cell viability assays. The potent ginsenoside, PPD was investigated for its roles in lipid raft disruption and downstream pathways to apoptosis of cancer cells. Anti-cancer effects of PPD was also investigated in vivo using mouse xenograft model. Results PPD consistently exerts its potent cytotoxicity in 2 cell survival assays using 5 different cancer cell lines. PPD disrupts lipid rafts in different ways from methyl-β-cyclodextrin (MβCD) depleting cholesterol out of the subdomains, since lipid raft proteins were differentially modulated by the saponin. During disruption of lipid rafts, PPD activated neutral sphingomyelinase 2 (nSMase 2) hydrolyzing membrane sphingomyelins into pro-apoptotic intracellular ceramides. Furthermore, PPD demonstrated its anti-cancer activities against K562 tumor cells in mouse xenograft model, confirming its potential as an adjunct or chemotherapeutic agent by itself in vivo. Conclusions This study demonstrates that neutral sphingomyelinase 2 is responsible for the cytotoxicity of PPD through production of apoptotic ceramides from membrane sphingomyelins. Thus neutral sphingomyelinase 2 and its relevant mechanisms may potentially be employed in cancer chemotherapies. PMID:23889969

  10. Sphingomyelinase D Activity in Model Membranes: Structural Effects of in situ Generation of Ceramide-1-Phosphate

    PubMed Central

    Stock, Roberto P.; Brewer, Jonathan; Wagner, Kerstin; Ramos-Cerrillo, Blanca; Duelund, Lars; Jernshøj, Kit Drescher; Olsen, Lars Folke; Bagatolli, Luis A.

    2012-01-01

    The toxicity of Loxosceles spider venom has been attributed to a rare enzyme, sphingomyelinase D, which transforms sphingomyelin to ceramide-1-phosphate. The bases of its inflammatory and dermonecrotic activity, however, remain unclear. In this work the effects of ceramide-1-phosphate on model membranes were studied both by in situ generation of this lipid using a recombinant sphingomyelinase D from the spider Loxosceles laeta and by pre-mixing it with sphingomyelin and cholesterol. The systems of choice were large unilamellar vesicles for bulk studies (enzyme kinetics, fluorescence spectroscopy and dynamic light scattering) and giant unilamellar vesicles for fluorescence microscopy examination using a variety of fluorescent probes. The influence of membrane lateral structure on the kinetics of enzyme activity and the consequences of enzyme activity on the structure of target membranes containing sphingomyelin were examined. The findings indicate that: 1) ceramide-1-phosphate (particularly lauroyl ceramide-1-phosphate) can be incorporated into sphingomyelin bilayers in a concentration-dependent manner and generates coexistence of liquid disordered/solid ordered domains, 2) the activity of sphingomyelinase D is clearly influenced by the supramolecular organization of its substrate in membranes and, 3) in situ ceramide-1-phosphate generation by enzymatic activity profoundly alters the lateral structure and morphology of the target membranes. PMID:22558302

  11. Role of Inositol Phosphosphingolipid Phospholipase C1, the Yeast Homolog of Neutral Sphingomyelinases in DNA Damage Response and Diseases.

    PubMed

    Tripathi, Kaushlendra

    2015-01-01

    Sphingolipids play a very crucial role in many diseases and are well-known as signaling mediators in many pathways. Sphingolipids are produced during the de novo process in the ER (endoplasmic reticulum) from the nonsphingolipid precursor and comprise both structural and bioactive lipids. Ceramide is the central core of the sphingolipid pathway, and its production has been observed following various treatments that can induce several different cellular effects including growth arrest, DNA damage, apoptosis, differentiation, and senescence. Ceramides are generally produced through the sphingomyelin hydrolysis and catalyzed by the enzyme sphingomyelinase (SMase) in mammals. Presently, there are many known SMases and they are categorized into three groups acid SMases (aSMases), alkaline SMases (alk-SMASES), and neutral SMases (nSMases). The yeast homolog of mammalians neutral SMases is inositol phosphosphingolipid phospholipase C. Yeasts generally have inositol phosphosphingolipids instead of sphingomyelin, which may act as a homolog of mammalian sphingomyelin. In this review, we shall explain the structure and function of inositol phosphosphingolipid phospholipase C1, its localization inside the cells, mechanisms, and its roles in various cell responses during replication stresses and diseases. This review will also give a new basis for our understanding for the mechanisms and nature of the inositol phosphosphingolipid phospholipase C1/nSMase. PMID:26346287

  12. Age‐related remodeling of small arteries is accompanied by increased sphingomyelinase activity and accumulation of long‐chain ceramides

    PubMed Central

    Ohanian, Jacqueline; Liao, Aiyin; Forman, Simon P.; Ohanian, Vasken

    2014-01-01

    Abstract The structure and function of large arteries alters with age leading to increased risk of cardiovascular disease. Age‐related large artery remodeling and arteriosclerosis is associated with increased collagen deposition, inflammation, and endothelial dysfunction. Bioactive sphingolipids are known to regulate these processes, and are also involved in aging and cellular senescence. However, less is known about age‐associated alterations in small artery morphology and function or whether changes in arterial sphingolipids occur in aging. We show that mesenteric small arteries from old sheep have increased lumen diameter and media thickness without a change in media to lumen ratio, indicative of outward hypertrophic remodeling. This remodeling occurred without overt changes in blood pressure or pulse pressure indicating it was a consequence of aging per se. There was no age‐associated change in mechanical properties of the arteries despite an increase in total collagen content and deposition of collagen in a thickened intima layer in arteries from old animals. Analysis of the sphingolipid profile showed an increase in long‐chain ceramide (C14–C20), but no change in the levels of sphingosine or sphingosine‐1‐phosphate in arteries from old compared to young animals. This was accompanied by a parallel increase in acid and neutral sphingomyelinase activity in old arteries compared to young. This study demonstrates remodeling of small arteries during aging that is accompanied by accumulation of long‐chain ceramides. This suggests that sphingolipids may be important mediators of vascular aging. PMID:24872355

  13. Neutral sphingomyelinase 2 is required for cytokine-induced skeletal muscle calpain activation.

    PubMed

    Supinski, Gerald S; Alimov, Alexander P; Wang, Lin; Song, Xiao-Hong; Callahan, Leigh A

    2015-09-15

    Calpain contributes to infection-induced diaphragm dysfunction but the upstream mechanism(s) responsible for calpain activation are poorly understood. It is known, however, that cytokines activate neutral sphingomyelinase (nSMase) and nSMase has downstream effects with the potential to increase calpain activity. We tested the hypothesis that infection-induced skeletal muscle calpain activation is a consequence of nSMase activation. We administered cytomix (20 ng/ml TNF-α, 50 U/ml IL-1β, 100 U/ml IFN-γ, 10 μg/ml LPS) to C2C12 muscle cells to simulate the effects of infection in vitro and studied mice undergoing cecal ligation puncture (CLP) as an in vivo model of infection. In cell studies, we assessed sphingomyelinase activity, subcellular calcium levels, and calpain activity and determined the effects of inhibiting sphingomyelinase using chemical (GW4869) and genetic (siRNA to nSMase2 and nSMase3) techniques. We assessed diaphragm force and calpain activity and utilized GW4869 to inhibit sphingomyelinase in mice. Cytomix increased cytosolic and mitochondrial calcium levels in C2C12 cells (P < 0.001); addition of GW4869 blocked these increases (P < 0.001). Cytomix also activated calpain, increasing calpain activity (P < 0.02), and the calpain-mediated cleavage of procaspase 12 (P < 0.001). Procaspase 12 cleavage was attenuated by either GW4869 (P < 0.001), BAPTA-AM (P < 0.001), or siRNA to nSMase2 (P < 0.001) but was unaffected by siRNA to nSMase3. GW4869 prevented CLP-induced diaphragm calpain activation and diaphragm weakness in mice. These data suggest that nSMase2 activation is required for the development of infection-induced diaphragm calpain activation and muscle weakness. As a consequence, therapies that inhibit nSMase2 in patients may prevent infection-induced skeletal muscle dysfunction. PMID:26138644

  14. Risk analysis of nuclear safeguards regulations. [Aggregated Systems Model (ASM)

    SciTech Connect

    Al-Ayat, R.A.; Altman, W.D.; Judd, B.R.

    1982-06-01

    The Aggregated Systems Model (ASM), a probabilisitic risk analysis tool for nuclear safeguards, was applied to determine benefits and costs of proposed amendments to NRC regulations governing nuclear material control and accounting systems. The objective of the amendments was to improve the ability to detect insiders attempting to steal large quantities of special nuclear material (SNM). Insider threats range from likely events with minor consequences to unlikely events with catastrophic consequences. Moreover, establishing safeguards regulations is complicated by uncertainties in threats, safeguards performance, and consequences, and by the subjective judgments and difficult trade-offs between risks and safeguards costs. The ASM systematically incorporates these factors in a comprehensive, analytical framework. The ASM was used to evaluate the effectiveness of current safeguards and to quantify the risk of SNM theft. Various modifications designed to meet the objectives of the proposed amendments to reduce that risk were analyzed. Safeguards effectiveness was judged in terms of the probability of detecting and preventing theft, the expected time to detection, and the expected quantity of SNM diverted in a year. Data were gathered in tours and interviews at NRC-licensed facilities. The assessment at each facility was begun by carefully selecting scenarios representing the range of potential insider threats. A team of analysts and facility managers assigned probabilities for detection and prevention events in each scenario. Using the ASM we computed the measures of system effectiveness and identified cost-effective safeguards modifications that met the objectives of the proposed amendments.

  15. Biochemical identification of a neutral sphingomyelinase 1 (NSM1)-like enzyme as the major NSM activity in the DT40 B-cell line: absence of a role in the apoptotic response to endoplasmic reticulum stress.

    PubMed Central

    Fensome, Amanda C; Josephs, Michelle; Katan, Matilda; Rodrigues-Lima, Fernando

    2002-01-01

    DT40 cells have approx. 10-fold higher Mg2+-dependent neutral sphingomyelinase (NSM) activity in comparison with other B-cell lines and contain very low acidic sphingomyelinase activity. Purification of this activity from DT40 cell membranes suggested the presence of one major NSM isoform. Although complete purification of this isoform could not be achieved, partially purified fractions were examined further with regard to the known characteristics of previously partially purified NSMs and the two cloned enzymes exhibiting in vitro NSM activity (NSM1 and NSM2). For a direct comparative study, highly purified brain preparations, purified NSM1 protein and Bacillus cereus enzyme were used. Analysis of the enzymic properties of the partially purified DT40 NSM, such as cation dependence, substrate specificity, redox regulation and stimulation by phosphatidylserine, together with the localization of this enzyme to the endoplasmic reticulum (ER), suggested that this NSM from DT40 cells corresponds to NSM1. Further studies aimed to correlate presence of the high levels of this NSM1-like activity in DT40 cells with the ability of these cells to accumulate ceramide and undergo apoptosis. When DT40 cells were stimulated to apoptose by a variety of agents, including the ER stress, an increase in endogenous ceramide levels was observed. However, these responses were not enhanced compared with another B-cell line (Nalm-6), characterized by low sphingomyelinase activity. In addition, DT40 cells were not more susceptible to ceramide accumulation and apoptosis when exposed to the ER stress compared with other apoptotic agents. Inhibition of de novo synthesis of ceramide partially inhibited its accumulation, indicating that the ceramide production in DT40 cells could be complex and, under some conditions, could involve both sphingomyelin hydrolysis and ceramide synthesis. PMID:12071841

  16. WERITAS: weighted ensemble of regional image textures for ASM segmentation

    NASA Astrophysics Data System (ADS)

    Toth, Robert; Doyle, Scott; Rosen, Mark; Kalyanpur, Arjun; Pungavkar, Sona; Bloch, B. Nicolas; Genega, Elizabeth; Rofsky, Neil; Lenkinski, Robert; Madabhushi, Anant

    2009-02-01

    In this paper we present WERITAS, which is based in part on the traditional Active Shape Model (ASM) segmentation system. WERITAS generates multiple statistical texture features, and finds the optimal weighted average of those texture features by maximizing the correlation between the Euclidean distance to the ground truth and the Mahalanobis distance to the training data. The weighted average is used a multi-resolution segmentation system to more accurately detect the object border. A rigorous evaluation was performed on over 200 clinical images comprising of prostate images and breast images from 1.5 Tesla and 3 Tesla MRI machines via 6 distinct metrics. WERITAS was tested against a traditional multi-resolution ASM in addition to an ASM system which uses a plethora of random features to determine if the selection of features is improving the results rather than simply the use of multiple features. The results indicate that WERITAS outperforms all other methods to a high degree of statistical significance. For 1.5T prostate MRI images, the overlap from WERITAS is 83%, the overlap from the random features is 81%, and the overlap from the traditional ASM is only 66%. In addition, using 3T prostate MRI images, the overlap from WERITAS is 77%, the overlap from the random features is 54%, and the overlap from the traditional ASM is 59%, suggesting the usefulness of WERITAS. The only metrics in which WERITAS was outperformed did not hold any degree of statistical significance. WERITAS is a robust, efficient, and accurate segmentation system with a wide range of applications.

  17. Operating nuclear plant feedback to ASME and French codes

    SciTech Connect

    Journet, J.; O`Donnell, W.J.

    1996-12-01

    The French have an advantage in nuclear plant operating experience feedback due to the highly centralized nature of their nuclear industry. There is only one utility in charge of design as well as operations (EDF) and only one reactor vendor (Framatome). The ASME Code has played a key role in resolving technical issues in the design and operation of nuclear plants since the inception of nuclear power. The committee structure of the Code brings an ideal combination of senior technical people with both broad and specialized experience to bear on complex how safe is safe enough technical issues. The authors now see an even greater role for the ASME Code in a proposed new regulatory era for the US nuclear industry. The current legalistic confrontational regulatory era has been quite destructive. There now appears to be a real opportunity to begin a new era of technical consensus as the primary means for resolving safety issues. This change can quickly be brought about by having the industry take operating plant problems and regulatory technical issues directly to the ASME Code for timely resolution. Surprisingly, there is no institution in the US nuclear industry with such a mandate. In fact, the industry is organized to feedback through the Nuclear Regulatory Commission issues which could be far better resolved through the ASME Code. Major regulatory benefits can be achieved by closing this loop and providing systematic interaction with the ASME Code. The essential elements of a new regulatory era and ideas for organizing US institutional industry responsibilities, taken from the French experience, are described in this paper.

  18. 75 FR 24323 - American Society of Mechanical Engineers (ASME) Codes and New and Revised ASME Code Cases

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-05-04

    ... by reference into the regulations in a final rule dated September 10, 2008 (73 FR 52730), as corrected on October 2, 2008 (73 FR 57235), incorporating Section III of the 2004 Edition of the ASME B&PV..., 2008 (73 FR 52729). The NRC follows a three-step process to determine acceptability of new...

  19. Molecular Evolution, Functional Variation, and Proposed Nomenclature of the Gene Family That Includes Sphingomyelinase D in Sicariid Spider Venoms

    PubMed Central

    Bodner, Melissa R.; Cordes, Matthew H.J.; Baldwin, Katherine L.; Rynerson, Melody R.; Burns, Scott N.; Zobel-Thropp, Pamela A.

    2009-01-01

    The venom enzyme sphingomyelinase D (SMase D) in the spider family Sicariidae (brown or fiddleback spiders [Loxosceles] and six-eyed sand spiders [Sicarius]) causes dermonecrosis in mammals. SMase D is in a gene family with multiple venom-expressed members that vary in functional specificity. We analyze molecular evolution of this family and variation in SMase D activity among crude venoms using a data set that represents the phylogenetic breadth of Loxosceles and Sicarius. We isolated a total of 190 nonredundant nucleotide sequences encoding 168 nonredundant amino acid sequences of SMase D homologs from 21 species. Bayesian phylogenies support two major clades that we name α and β, within which we define seven and three subclades, respectively. Sequences in the α clade are exclusively from New World Loxosceles and Loxosceles rufescens and include published genes for which expression products have SMase D and dermonecrotic activity. The β clade includes paralogs from New World Loxosceles that have no, or reduced, SMase D and no dermonecrotic activity and also paralogs from Sicarius and African Loxosceles of unknown activity. Gene duplications are frequent, consistent with a birth-and-death model, and there is evidence of purifying selection with episodic positive directional selection. Despite having venom-expressed SMase D homologs, venoms from New World Sicarius have reduced, or no, detectable SMase D activity, and Loxosceles in the Southern African spinulosa group have low SMase D activity. Sequence conservation mapping shows >98% conservation of proposed catalytic residues of the active site and around a plug motif at the opposite end of the TIM barrel, but α and β clades differ in conservation of key residues surrounding the apparent substrate binding pocket. Based on these combined results, we propose an inclusive nomenclature for the gene family, renaming it SicTox, and discuss emerging patterns of functional diversification. PMID:19042943

  20. New oleyl glycoside as anti-cancer agent that targets on neutral sphingomyelinase.

    PubMed

    Romero-Ramírez, Lorenzo; García-Álvarez, Isabel; Casas, Josefina; Barreda-Manso, M Asunción; Yanguas-Casás, Natalia; Nieto-Sampedro, Manuel; Fernández-Mayoralas, Alfonso

    2015-09-15

    We designed and synthesized two anomeric oleyl glucosaminides as anti-cancer agents where the presence of a trifluoroacetyl group close to the anomeric center makes them resistant to hydrolysis by hexosaminidases. The oleyl glycosides share key structural features with synthetic and natural oleyl derivatives that have been reported to exhibit anti-cancer properties. While both glycosides showed antiproliferative activity on cancer cell lines, only the α-anomer caused endoplasmic reticulum (ER) stress and cell death on C6 glioma cells. Analysis of sphingolipids and glycosphingolipds in cells treated with the glycosides showed that the α-anomer caused a drastic accumulation of ceramide and glucosylceramide and reduction of lactosylceramide and GM3 ganglioside at concentrations above a threshold of 20 μM. In order to understand how ceramide levels increase in response to α-glycoside treatment, further investigations were done using specific inhibitors of sphingolipid metabolic pathways. The pretreatment with 3-O-methylsphingomyelin (a neutral sphingomyelinase inhibitor) restored sphingomyelin levels together with the lactosylceramide and GM3 ganglioside levels and prevented the ER stress and cell death caused by the α-glycoside. The results indicated that the activation of neutral sphingomyelinase is the main cause of the alterations in sphingolipids that eventually lead to cell death. The new oleyl glycoside targets a key enzyme in sphingolipid metabolism with potential applications in cancer therapy. PMID:26206186

  1. Mechanism of induction of complement susceptibility of erythrocytes by spider and bacterial sphingomyelinases

    PubMed Central

    Tambourgi, Denise V; De Sousa Da Silva, Marcelo; Billington, Stephen J; Gonçalves De Andrade, Rute M; Magnoli, Fábio C; Songer, J Glenn; Van Den Berg, Carmen W

    2002-01-01

    We have recently shown that the sphingomyelinase toxins P1 and P2 from the venom of the spider Loxosceles intermedia induce complement (C)-dependent lysis of autologous erythrocytes by induction of the cleavage of cell surface glycophorins through activation of an endogenous metalloproteinase facilitating the activation of the alternative pathway of C. Phospholipase D (PLD) from Corynebacterium pseudotuberculosis shows some degree of homology with the spider sphingomyelinases and can induce similar clinical symptoms to those observed after spider envenomation. The aim of this study was to investigate if the bacterial PLD-induced haemolysis of human erythrocytes was C dependent and if cleavage of glycophorins occurred. We show here that haemolysis of both PLD- and P1-treated human erythrocytes was C dependent, but while PLD-mediated haemolysis was dependent on activation of the classical pathway of C, P1 induced lysis via both the classical and alternative pathways. P1, but not PLD, induced cleavage of glycophorins and no change in expression of complement regulators was induced by either of the toxins. In both cases, annexin V binding sites were exposed, suggesting that the membrane asymmetry had been disturbed causing exposure of phosphatidylserine to the cell surface. Our results suggest that C susceptibility induced by L. intermedia and C. pseudotuberculosis PLD is a result of exposure of phosphatidylserine, and the higher potency of P1 toxin can be explained by its additional effect of cleavage of glycophorins. PMID:12225367

  2. Neutral Sphingomyelinase in Physiological and Measles Virus Induced T Cell Suppression

    PubMed Central

    Collenburg, Lena; Grassmé, Heike; Schneider-Schaulies, Sibylle

    2014-01-01

    T cell paralysis is a main feature of measles virus (MV) induced immunosuppression. MV contact mediated activation of sphingomyelinases was found to contribute to MV interference with T cell actin reorganization. The role of these enzymes in MV-induced inhibition of T cell activation remained equally undefined as their general role in regulating immune synapse (IS) activity which relies on spatiotemporal membrane patterning. Our study for the first time reveals that transient activation of the neutral sphingomyelinase 2 (NSM2) occurs in physiological co-stimulation of primary T cells where ceramide accumulation is confined to the lamellum (where also NSM2 can be detected) and excluded from IS areas of high actin turnover. Genetic ablation of the enzyme is associated with T cell hyper-responsiveness as revealed by actin dynamics, tyrosine phosphorylation, Ca2+-mobilization and expansion indicating that NSM2 acts to suppress overshooting T cell responses. In line with its suppressive activity, exaggerated, prolonged NSM2 activation as occurring in co-stimulated T cells following MV exposure was associated with aberrant compartmentalization of ceramides, loss of spreading responses, interference with accumulation of tyrosine phosphorylated protein species and expansion. Altogether, this study for the first time reveals a role of NSM2 in physiological T cell stimulation which is dampening and can be abused by a virus, which promotes enhanced and prolonged NSM2 activation to cause pathological T cell suppression. PMID:25521388

  3. Recent changes to ASME B31.3

    SciTech Connect

    Becht, C. IV; Frikken, D.R.; Bane, E.J.

    1996-07-01

    The code for process piping, ASME B31.3 Chemical Plant and Petroleum Refinery Piping, has undergone significant changes and additions in recent years. This includes many aspects of design, materials, and fabrication. Included are substantial changes to material impact testing requirements, qualification procedures for unlisted components, coverage of bellows expansion joints, and safety relief set pressure requirements. This paper provides an update on some of these recent changes to the Code together with some background on reasons for the changes.

  4. ASME code post weld heat treatment requirements: Current issues

    SciTech Connect

    McEnerney, J.W.

    1995-12-31

    An understanding of current ASME Code PWHT issues requires consideration of various governing factors. These include: historical perspective regarding rule development and needs for PWHT, design criteria dictating the purpose, potential adverse effects, validity of existing and/or need for additional global PWHT parameters, and fundamental questions and/or inadequate current requirements regarding local PWHT. Various activities to address the current issues are in progress at different code task groups and organizations.

  5. Proceedings of the 1994 ASME/IEEE joint railroad conference

    SciTech Connect

    Hawthorne, K.L.; Hill, J.R.

    1994-12-31

    The proceedings contains 22 papers from the IEEE/ASME Joint Railroad Conference. Topics discussed include flange bearing crossing frogs; rapid transit; vehicle on-board computer systems; total transport system control functions; automated train dispatching system; wheel thermal damage limits; AC propulsion; AC traction drives; electric power supplies; failure detection and identification; cargo transportation; and critical speed for railroad vehicles. Papers within the scope of the data base have been processed separately.

  6. Sphingomyelinase promotes oxidant production and skeletal muscle contractile dysfunction through activation of NADPH oxidase

    PubMed Central

    Loehr, James A.; Abo-Zahrah, Reem; Pal, Rituraj; Rodney, George G.

    2015-01-01

    Elevated concentrations of sphingomyelinase (SMase) have been detected in a variety of diseases. SMase has been shown to increase muscle derived oxidants and decrease skeletal muscle force; however, the sub-cellular site of oxidant production has not been elucidated. Using redox sensitive biosensors targeted to the mitochondria and NADPH oxidase (Nox2), we demonstrate that SMase increased Nox2-dependent ROS and had no effect on mitochondrial ROS in isolated FDB fibers. Pharmacological inhibition and genetic knockdown of Nox2 activity prevented SMase induced ROS production and provided protection against decreased force production in the diaphragm. In contrast, genetic overexpression of superoxide dismutase within the mitochondria did not prevent increased ROS production and offered no protection against decreased diaphragm function in response to SMase. Our study shows that SMase induced ROS production occurs in specific sub-cellular regions of skeletal muscle; however, the increased ROS does not completely account for the decrease in muscle function. PMID:25653619

  7. Roles and regulation of neutral sphingomyelinase-2 in cellular and pathological processes.

    PubMed

    Shamseddine, Achraf A; Airola, Michael V; Hannun, Yusuf A

    2015-01-01

    Our understanding of the functions of ceramide signaling has advanced tremendously over the past decade. In this review, we focus on the roles and regulation of neutral sphingomyelinase 2 (nSMase2), an enzyme that generates the bioactive lipid ceramide through the hydrolysis of the membrane lipid sphingomyelin. A large body of work has now implicated nSMase2 in a diverse set of cellular functions, physiological processes, and disease pathologies. We discuss different aspects of this enzyme's regulation from transcriptional, post-translational, and biochemical. Furthermore, we highlight nSMase2 involvement in cellular processes including inflammatory signaling, exosome generation, cell growth, and apoptosis, which in turn play important roles in pathologies such as cancer metastasis, Alzheimer's disease, and other organ systems disorders. Lastly, we examine avenues where targeted nSMase2-inhibition may be clinically beneficial in disease scenarios. PMID:25465297

  8. Roles and regulation of Neutral Sphingomyelinase-2 in cellular and pathological processes

    PubMed Central

    Shamseddine, Achraf A.; Airola, Michael V.; Hannun, Yusuf A.

    2015-01-01

    Our understanding of the functions of ceramide signaling has advanced tremendously over the past decade. In this review, we focus on the roles and regulation of neutral sphingomyelinase 2 (nSMase2), an enzyme that generates the bioactive lipid ceramide through the hydrolysis of the membrane lipid sphingomyelin. A large body of work has now implicated nSMase2 in a diverse set of cellular functions, physiological processes, and disease pathologies. We discuss different aspects of this enzyme’s regulation from transcriptional, post-translational, and biochemical. Furthermore, we highlight nSMase2 involvement in cellular processes including inflammatory signaling, exosome generation, cell growth, and apoptosis, which in turn play important roles in pathologies such as cancer metastasis, Alzheimer’s disease, and other organ systems disorders. Lastly, we examine avenues where targeted nSMase2-inhibition may be clinically beneficial in disease scenarios. PMID:25465297

  9. Src regulates cigarette smoke-induced ceramide generation via neutral sphingomyelinase 2 in the airway epithelium.

    PubMed

    Chung, Samuel; Vu, Simon; Filosto, Simone; Goldkorn, Tzipora

    2015-06-01

    We previously demonstrated that the neutral sphingomyelinase (nSMase) 2 is the sole sphingomyelinase activated during cigarette smoke (CS)-induced oxidative stress of human airway epithelial cells, leading to ceramide generation and subsequent apoptosis of affected cells. Since then, we reported that nSMase2 is a phosphoprotein, the degree of enzymatic activity and stability of which are dictated by its degree of phosphorylation. Simultaneously, the non-receptor tyrosine kinase and proto-oncogene Src has increasingly become a target of interest in both smoking-related lung injury, such as chronic obstructive pulmonary disease, and lung cancer. Within this context, we tested and now present Src as a regulator of ceramide generation via modulation of nSMase2 phosphorylation and activity during CS-induced oxidative stress. Specifically, we provide evidence that Src activity is necessary for both CS-induced ceramide accumulation in vivo (129/Sv mice) and in vitro (human airway epithelial cells) and for nSMase2 activity during CS-induced oxidative stress. Moreover, because nSMase2 is exclusively phosphorylated on serines, we show that this occurs through Src-dependent activation of the serine/threonine kinase p38 mitogen-activated protein kinase during oxidative stress. Finally, we provide evidence that Src and p38 mitogen-activated protein kinase activities are critical for regulating nSMase2 phosphorylation. This study provides insights into a molecular target involved in smoking-related lung injury, represented here as nSMase2, and its modulation by the oncogene Src. PMID:25347576

  10. Src Regulates Cigarette Smoke–Induced Ceramide Generation via Neutral Sphingomyelinase 2 in the Airway Epithelium

    PubMed Central

    Chung, Samuel; Vu, Simon; Filosto, Simone

    2015-01-01

    We previously demonstrated that the neutral sphingomyelinase (nSMase) 2 is the sole sphingomyelinase activated during cigarette smoke (CS)–induced oxidative stress of human airway epithelial cells, leading to ceramide generation and subsequent apoptosis of affected cells. Since then, we reported that nSMase2 is a phosphoprotein, the degree of enzymatic activity and stability of which are dictated by its degree of phosphorylation. Simultaneously, the non–receptor tyrosine kinase and proto-oncogene Src has increasingly become a target of interest in both smoking-related lung injury, such as chronic obstructive pulmonary disease, and lung cancer. Within this context, we tested and now present Src as a regulator of ceramide generation via modulation of nSMase2 phosphorylation and activity during CS-induced oxidative stress. Specifically, we provide evidence that Src activity is necessary for both CS-induced ceramide accumulation in vivo (129/Sv mice) and in vitro (human airway epithelial cells) and for nSMase2 activity during CS-induced oxidative stress. Moreover, because nSMase2 is exclusively phosphorylated on serines, we show that this occurs through Src-dependent activation of the serine/threonine kinase p38 mitogen-activated protein kinase during oxidative stress. Finally, we provide evidence that Src and p38 mitogen-activated protein kinase activities are critical for regulating nSMase2 phosphorylation. This study provides insights into a molecular target involved in smoking-related lung injury, represented here as nSMase2, and its modulation by the oncogene Src. PMID:25347576

  11. Cambinol, a Novel Inhibitor of Neutral Sphingomyelinase 2 Shows Neuroprotective Properties

    PubMed Central

    Figuera-Losada, Mariana; Stathis, Marigo; Dorskind, Joelle M.; Thomas, Ajit G.; Bandaru, Veera Venkata Ratnam; Yoo, Seung-Wan; Westwood, Nicholas J.; Rogers, Graeme W.; McArthur, Justin C.; Haughey, Norman J.; Slusher, Barbara S.; Rojas, Camilo

    2015-01-01

    Ceramide is a bioactive lipid that plays an important role in stress responses leading to apoptosis, cell growth arrest and differentiation. Ceramide production is due in part to sphingomyelin hydrolysis by sphingomyelinases. In brain, neutral sphingomyelinase 2 (nSMase2) is expressed in neurons and increases in its activity and expression have been associated with pro-inflammatory conditions observed in Alzheimer’s disease, multiple sclerosis and human immunodeficiency virus (HIV-1) patients. Increased nSMase2 activity translates into higher ceramide levels and neuronal cell death, which can be prevented by chemical or genetic inhibition of nSMase2 activity or expression. However, to date, there are no soluble, specific and potent small molecule inhibitor tool compounds for in vivo studies or as a starting point for medicinal chemistry optimization. Moreover, the majority of the known inhibitors were identified using bacterial, bovine or rat nSMase2. In an attempt to identify new inhibitor scaffolds, two activity assays were optimized as screening platform using the recombinant human enzyme. First, active hits were identified using a fluorescence-based high throughput compatible assay. Then, hits were confirmed using a 14C sphingomyelin-based direct activity assay. Pharmacologically active compounds and approved drugs were screened using this strategy which led to the identification of cambinol as a novel uncompetitive nSMase2 inhibitor (Ki = 7 μM). The inhibitory activity of cambinol for nSMase2 was approximately 10-fold more potent than for its previously known target, silence information regulator 1 and 2 (SIRT1/2). Cambinol decreased tumor necrosis factor-α or interleukin-1 β-induced increases of ceramide and cell death in primary neurons. A preliminary study of cambinol structure and activity allowed the identification of the main structural features required for nSMase2 inhibition. Cambinol and its analogs may be useful as nSMase2 inhibitor tool

  12. ASME PTC 46 -- Acceptance test code for overall plant performance

    SciTech Connect

    Friedman, J.R.; Yost, J.G.

    1999-11-01

    ASME published PTC 46 in 1996 after five years of development. PTC 46 is the first industry standard providing explicit procedures for conducting acceptance tests to determine the overall thermal performance and output of power generating units. It is applicable to any heat cycle power generating unit. This survey paper provides an overview of PTC 46 and discusses how PTC 46 can be used for acceptance testing of new combined cycle and fossil steam power generating units. Several technical papers have been previously presented that provide more detailed information and discussion on the use of PTC 46 in acceptance testing.

  13. Significant issues and changes for ANSI/ASME OM-1 1981, part 1, ASME OMc code-1994, and ASME OM Code-1995, Appendix I, inservice testing of pressure relief devices in light water reactor power plants

    SciTech Connect

    Seniuk, P.J.

    1996-12-01

    This paper identifies significant changes to the ANSI/ASME OM-1 1981, Part 1, and ASME Omc Code-1994 and ASME OM Code-1995, Appendix I, {open_quotes}Inservice Testing of Pressure Relief Devices in Light-Water Reactor Power Plants{close_quotes}. The paper describes changes to different Code editions and presents insights into the direction of the code committee and selected topics to be considered by the ASME O&M Working Group on pressure relief devices. These topics include scope issues, thermal relief valve issues, as-found and as-left set-pressure determinations, exclusions from testing, and cold setpoint bench testing. The purpose of this paper is to describe some significant issues being addressed by the O&M Working Group on Pressure Relief Devices (OM-1). The writer is currently the chair of OM-1 and the statements expressed herein represents his personal opinion.

  14. Purification and characterization of two Listeria ivanovii cytolysins, a sphingomyelinase C and a thiol-activated toxin (ivanolysin O).

    PubMed Central

    Vazquez-Boland, J A; Dominguez, L; Rodriguez-Ferri, E F; Suarez, G

    1989-01-01

    The strong bizonal hemolysis on blood agar and the positive CAMP reaction with Rhodococcus equi denotes the production of two different cytolytic factors by Listeria ivanovii. One was characterized as a thiol-activated (SH) cytolysin of 61 kilodaltons and was termed ivanolysin O (ILO) since data suggested that it is different from listeriolysin O, the SH-cytolysin produced by Listeria monocytogenes. The other is a 27-kilodalton hemolytic sphingomyelinase C that was found to be the cytolytic factor responsible for the halo of incomplete hemolysis synergistically enhanced by R. equi exosubstances. When thiol-disulfide exchange affinity chromatography and gel filtration were applied to the purification of ILO from concentrated L. ivanovii culture supernatants, the copurification of the two cytolysins was observed. This phenomenon seems to be due to the formation of intermolecular disulfide bonds between ILO and the sphingomyelinase, since the latter was found to contain free SH groups, not essential for the activity. These SH groups could react with the single cysteine residue characteristically present in the SH-cytolysins, forming a dimeric cytolytic complex. The purification of ILO was achieved by a further gel filtration with a reducing agent (dithiothreitol) in the eluent. A method for the purification of the sphingomyelinase based on selective sequestration of ILO from the L. ivanovii concentrated culture supernatant by the SH cytolysin target molecule cholesterol and thiol-disulfide affinity chromatography is described. Images PMID:2553614

  15. ASM Based Synthesis of Handwritten Arabic Text Pages.

    PubMed

    Dinges, Laslo; Al-Hamadi, Ayoub; Elzobi, Moftah; El-Etriby, Sherif; Ghoneim, Ahmed

    2015-01-01

    Document analysis tasks, as text recognition, word spotting, or segmentation, are highly dependent on comprehensive and suitable databases for training and validation. However their generation is expensive in sense of labor and time. As a matter of fact, there is a lack of such databases, which complicates research and development. This is especially true for the case of Arabic handwriting recognition, that involves different preprocessing, segmentation, and recognition methods, which have individual demands on samples and ground truth. To bypass this problem, we present an efficient system that automatically turns Arabic Unicode text into synthetic images of handwritten documents and detailed ground truth. Active Shape Models (ASMs) based on 28046 online samples were used for character synthesis and statistical properties were extracted from the IESK-arDB database to simulate baselines and word slant or skew. In the synthesis step ASM based representations are composed to words and text pages, smoothed by B-Spline interpolation and rendered considering writing speed and pen characteristics. Finally, we use the synthetic data to validate a segmentation method. An experimental comparison with the IESK-arDB database encourages to train and test document analysis related methods on synthetic samples, whenever no sufficient natural ground truthed data is available. PMID:26295059

  16. ASM Based Synthesis of Handwritten Arabic Text Pages

    PubMed Central

    Dinges, Laslo; Al-Hamadi, Ayoub; Elzobi, Moftah; El-etriby, Sherif; Ghoneim, Ahmed

    2015-01-01

    Document analysis tasks, as text recognition, word spotting, or segmentation, are highly dependent on comprehensive and suitable databases for training and validation. However their generation is expensive in sense of labor and time. As a matter of fact, there is a lack of such databases, which complicates research and development. This is especially true for the case of Arabic handwriting recognition, that involves different preprocessing, segmentation, and recognition methods, which have individual demands on samples and ground truth. To bypass this problem, we present an efficient system that automatically turns Arabic Unicode text into synthetic images of handwritten documents and detailed ground truth. Active Shape Models (ASMs) based on 28046 online samples were used for character synthesis and statistical properties were extracted from the IESK-arDB database to simulate baselines and word slant or skew. In the synthesis step ASM based representations are composed to words and text pages, smoothed by B-Spline interpolation and rendered considering writing speed and pen characteristics. Finally, we use the synthetic data to validate a segmentation method. An experimental comparison with the IESK-arDB database encourages to train and test document analysis related methods on synthetic samples, whenever no sufficient natural ground truthed data is available. PMID:26295059

  17. 3D automatic anatomy recognition based on iterative graph-cut-ASM

    NASA Astrophysics Data System (ADS)

    Chen, Xinjian; Udupa, Jayaram K.; Bagci, Ulas; Alavi, Abass; Torigian, Drew A.

    2010-02-01

    We call the computerized assistive process of recognizing, delineating, and quantifying organs and tissue regions in medical imaging, occurring automatically during clinical image interpretation, automatic anatomy recognition (AAR). The AAR system we are developing includes five main parts: model building, object recognition, object delineation, pathology detection, and organ system quantification. In this paper, we focus on the delineation part. For the modeling part, we employ the active shape model (ASM) strategy. For recognition and delineation, we integrate several hybrid strategies of combining purely image based methods with ASM. In this paper, an iterative Graph-Cut ASM (IGCASM) method is proposed for object delineation. An algorithm called GC-ASM was presented at this symposium last year for object delineation in 2D images which attempted to combine synergistically ASM and GC. Here, we extend this method to 3D medical image delineation. The IGCASM method effectively combines the rich statistical shape information embodied in ASM with the globally optimal delineation capability of the GC method. We propose a new GC cost function, which effectively integrates the specific image information with the ASM shape model information. The proposed methods are tested on a clinical abdominal CT data set. The preliminary results show that: (a) it is feasible to explicitly bring prior 3D statistical shape information into the GC framework; (b) the 3D IGCASM delineation method improves on ASM and GC and can provide practical operational time on clinical images.

  18. 46 CFR 57.02-2 - Adoption of section IX of the ASME Code.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 2 2010-10-01 2010-10-01 false Adoption of section IX of the ASME Code. 57.02-2 Section 57.02-2 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) MARINE ENGINEERING WELDING... accordance with section IX of the ASME (American Society of Mechanical Engineers) Code, as limited,...

  19. 46 CFR 57.02-2 - Adoption of section IX of the ASME Code.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 2 2013-10-01 2013-10-01 false Adoption of section IX of the ASME Code. 57.02-2 Section 57.02-2 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) MARINE ENGINEERING WELDING... accordance with section IX of the ASME (American Society of Mechanical Engineers) Code, as limited,...

  20. 46 CFR 57.02-2 - Adoption of section IX of the ASME Code.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 2 2012-10-01 2012-10-01 false Adoption of section IX of the ASME Code. 57.02-2 Section 57.02-2 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) MARINE ENGINEERING WELDING... accordance with section IX of the ASME (American Society of Mechanical Engineers) Code, as limited,...

  1. ASME Nuclear Crane Standards for Enhanced Crane Safety and Increased Profit

    NASA Astrophysics Data System (ADS)

    Parkhurst, Stephen N.

    2000-01-01

    The ASME NOG-1 standard, 'Rules for Construction of Overhead and Gantry Cranes', covers top running cranes for nuclear facilities; with the ASME NUM-1 standard, 'Rules for Construction of Cranes, Monorails, and Hoists', covering the single girder, underhung, wall and jib cranes, as well as the monorails and hoists. These two ASME nuclear crane standards provide criteria for designing, inspecting and testing overhead handling equipment with enhanced safety to meet the 'defense-in-depth' approach of the United States Nuclear Regulatory Commission (USNRC) documents NUREG 0554 and NUREG 0612. In addition to providing designs for enhanced safety, the ASME nuclear crane standards provide a basis for purchasing overhead handling equipment with standard safety features, based upon accepted engineering principles, and including performance and environmental parameters specific to nuclear facilities. The ASME NOG-1 and ASME NUM-1 standards not only provide enhanced safety for handling a critical load, but also increase profit by minimizing the possibility of load drops, by reducing cumbersome operating restrictions, and by providing the foundation for a sound licensing position. The ASME nuclear crane standards can also increase profit by providing the designs and information to help ensure that the right standard equipment is purchased. Additionally, the ASME nuclear crane standards can increase profit by providing designs and information to help address current issues, such as the qualification of nuclear plant cranes for making 'planned engineered lifts' for steam generator replacement and decommissioning.

  2. A Review & Assessment of Current Operating Conditions Allowable Stresses in ASME Section III Subsection NH

    SciTech Connect

    R. W. Swindeman

    2009-12-14

    The current operating condition allowable stresses provided in ASME Section III, Subsection NH were reviewed for consistency with the criteria used to establish the stress allowables and with the allowable stresses provided in ASME Section II, Part D. It was found that the S{sub o} values in ASME III-NH were consistent with the S values in ASME IID for the five materials of interest. However, it was found that 0.80 S{sub r} was less than S{sub o} for some temperatures for four of the materials. Only values for alloy 800H appeared to be consistent with the criteria on which S{sub o} values are established. With the intent of undertaking a more detailed evaluation of issues related to the allowable stresses in ASME III-NH, the availabilities of databases for the five materials were reviewed and augmented databases were assembled.

  3. Enhanced Sphingomyelinase Activity Contributes to the Apoptotic Capacity of Electronegative Low-Density Lipoprotein.

    PubMed

    Ke, Liang-Yin; Chan, Hua-Chen; Chen, Chih-Chieh; Lu, Jonathan; Marathe, Gopal K; Chu, Chih-Sheng; Chan, Hsiu-Chuan; Wang, Chung-Ya; Tung, Yi-Ching; McIntyre, Thomas M; Yen, Jeng-Hsien; Chen, Chu-Huang

    2016-02-11

    Sphingomyelinase (SMase) catalyzes the degradation of sphingomyelin to ceramide. In patients with metabolic syndrome or diabetes, circulating plasma ceramide levels are significantly higher than in normal individuals. Our data indicate that electronegative low-density lipoprotein (LDL) shows SMase activity, which leads to increased ceramide levels that can produce pro-inflammatory effects and susceptibility to aggregation. According to sequence alignment and protein structure predictions, the putative catalytic site of SMase activity is in the α2 region of apoB-100. To identify specific post-translational modifications of apoB100 near the catalytic region, we performed data-independent, parallel-fragmentation liquid chromatography/mass spectrometry (LC/MS(E)), followed by data analysis with ProteinLynx GlobalServer v2.4. Results showed that the serine of apoB100 in electronegative LDL was highly O-glycosylated, including S(1732), S(1959), S(2378), S(2408), and S(2429). These findings may support the changing of the α-helix/β-pleated sheets ratio in protein structure analysis. Further study is necessary to confirm the activation of SMase activity by electronegative LDL. PMID:26766134

  4. Inhibition of neutral sphingomyelinase-2 perturbs brain sphingolipid balance and spatial memory in mice

    PubMed Central

    Tabatadze, Nino; Savonenko, Alena; Song, Hongjun; Bandaru, Veera Venkata Ratnam; Chu, Michael; Haughey, Norman J.

    2010-01-01

    The sphingolipid ceramide is a bioactive signaling lipid that is thought to play important roles in modulating synaptic activity, in part by regulating the function of excitatory postsynaptic receptors. However, the molecular mechanisms by which ceramide exerts its effects on synaptic activity remain largely unknown. We recently demonstrated that a rapid generation of ceramide by neutral sphingomyelinase-2 (nSMase2; also known as sphingomyelin phosphodiesterase-3) played a key role in modulating excitatory postsynaptic currents by controlling the insertion and clustering of NMDA receptors (Wheeler et al. 2009). We now demonstrate that nSMase2 plays a role in memory. Inhibition of nSMase2 impaired spatial and episodic-like memory in mice. At the molecular level, inhibition of nSMase2 decreased ceramide, increased PSD-95, increased the number of AMPA receptors and altered the subunit composition of NMDA receptors. Our study identifies nSMase2 as an important component for efficient memory formation and underscores the importance of ceramide in regulating synaptic events related to learning and memory. PMID:20629193

  5. Primary cilia in stem cells and neural progenitors are regulated by neutral sphingomyelinase 2 and ceramide

    PubMed Central

    He, Qian; Wang, Guanghu; Wakade, Sushama; Dasgupta, Somsankar; Dinkins, Michael; Kong, Ji Na; Spassieva, Stefka D.; Bieberich, Erhard

    2014-01-01

    We show here that human embryonic stem (ES) and induced pluripotent stem cell–derived neuroprogenitors (NPs) develop primary cilia. Ciliogenesis depends on the sphingolipid ceramide and its interaction with atypical PKC (aPKC), both of which distribute to the primary cilium and the apicolateral cell membrane in NP rosettes. Neural differentiation of human ES cells to NPs is concurrent with a threefold elevation of ceramide—in particular, saturated, long-chain C16:0 ceramide (N-palmitoyl sphingosine) and nonsaturated, very long chain C24:1 ceramide (N-nervonoyl sphingosine). Decreasing ceramide levels by inhibiting ceramide synthase or neutral sphingomyelinase 2 leads to translocation of membrane-bound aPKC to the cytosol, concurrent with its activation and the phosphorylation of its substrate Aurora kinase A (AurA). Inhibition of aPKC, AurA, or a downstream target of AurA, HDAC6, restores ciliogenesis in ceramide-depleted cells. Of importance, addition of exogenous C24:1 ceramide reestablishes membrane association of aPKC, restores primary cilia, and accelerates neural process formation. Taken together, these results suggest that ceramide prevents activation of HDAC6 by cytosolic aPKC and AurA, which promotes acetylation of tubulin in primary cilia and, potentially, neural processes. This is the first report on the critical role of ceramide generated by nSMase2 in stem cell ciliogenesis and differentiation. PMID:24694597

  6. P53-dependent upregulation of neutral sphingomyelinase-2: role in doxorubicin-induced growth arrest

    PubMed Central

    Shamseddine, A A; Clarke, C J; Carroll, B; Airola, M V; Mohammed, S; Rella, A; Obeid, L M; Hannun, Y A

    2015-01-01

    Neutral sphingomyelinase-2 (nSMase2) is a ceramide-generating enzyme that has been implicated in growth arrest, apoptosis and exosome secretion. Although previous studies have reported transcriptional upregulation of nSMase2 in response to daunorubicin, through Sp1 and Sp3 transcription factors, the role of the DNA damage pathway in regulating nSMase2 remains unclear. In this study, we show that doxorubicin induces a dose-dependent induction of nSMase2 mRNA and protein with concomitant increases in nSMase activity and ceramide levels. Upregulation of nSMase2 was dependent on ATR, Chk1 and p53, thus placing it downstream of the DNA damage pathway. Moreover, overexpression of p53 was sufficient to transcriptionally induce nSMase2, without the need for DNA damage. DNA-binding mutants as well as acetylation mutants of p53 were unable to induce nSMase2, suggesting a role of nSMase2 in growth arrest. Moreover, knockdown of nSMase2 prevented doxorubicin-induced growth arrest. Finally, p53-induced nSMase2 upregulation appears to occur via a novel transcription start site upstream of exon 3. These results identify nSMase2 as a novel p53 target gene, regulated by the DNA damage pathway to induce cell growth arrest. PMID:26512957

  7. Bidirectional control of sphingomyelinase activity and surface topography in lipid monolayers.

    PubMed

    Fanani, María Laura; Härtel, Steffen; Oliveira, Rafael G; Maggio, Bruno

    2002-12-01

    Lipid lateral organization is increasingly found to modulate membrane-bound enzymes. We followed in real time the reaction course of sphingomyelin (SM) degradation by Bacillus cereus sphingomyelinase (SMase) of lipid monolayers by epifluorescence microscopy. There is evidence that formation of ceramide (Cer), a lipid second messenger, drives structural reorganization of membrane lipids. Our results provide visual evidence that SMase activity initially alters surface topography by inducing phase separation into condensed (Cer-enriched) and expanded (SM-enriched) domains. The Cer-enriched phase grows steadily as the reaction proceeds at a constant rate. The surface topography derived from the SMase-driven reaction was compared with, and found to differ from, that of premixed SM/Cer monolayers of the same lipid composition, indicating that substantial information content is stored depending on the manner in which the surface was generated. The long-range topographic changes feed back on the kinetics of Smase, and the onset of condensed-phase percolation is temporally correlated with a rapid drop of reaction rate. These observations reveal a bidirectional influence and communication between effects taking place at the local molecular level and the supramolecular organization. The results suggest a novel biocatalytic-topographic mechanism in which a surface enzymatic activity can influence the function of amphitropic proteins important for cell function. PMID:12496108

  8. Mutations in the neutral sphingomyelinase gene SMPD3 implicate the ceramide pathway in human leukemias.

    PubMed

    Kim, Woo Jae; Okimoto, Ross A; Purton, Louise E; Goodwin, Meagan; Haserlat, Sara M; Dayyani, Farshid; Sweetser, David A; McClatchey, Andrea I; Bernard, Olivier A; Look, A Thomas; Bell, Daphne W; Scadden, David T; Haber, Daniel A

    2008-05-01

    Ceramide is a lipid second messenger derived from the hydrolysis of sphingomyelin by sphingomyelinases (SMases) and implicated in diverse cellular responses, including growth arrest, differentiation, and apoptosis. Defects in the neutral SMase (nSMase) gene Smpd3, the primary regulator of ceramide biosynthesis, are responsible for developmental defects of bone; regulation of ceramide levels have been implicated in macrophage differentiation, but this pathway has not been directly implicated in human cancer. In a genomic screen for gene copy losses contributing to tumorigenesis in a mouse osteosarcoma model, we identified a somatic homozygous deletion specifically targeting Smpd3. Reconstitution of SMPD3 expression in mouse tumor cells lacking the endogenous gene enhanced tumor necrosis factor (TNF)-induced reduction of cell viability. Nucleotide sequencing of the highly conserved SMPD3 gene in a large panel of human cancers revealed mutations in 5 (5%) of 92 acute myeloid leukemias (AMLs) and 8 (6%) of 131 acute lymphoid leukemias (ALLs), but not in other tumor types. In a subset of these mutations, functional analysis indicated defects in protein stability and localization. Taken together, these observations suggest that disruption of the ceramide pathway may contribute to a subset of human leukemias. PMID:18299447

  9. Mutations in the neutral sphingomyelinase gene SMPD3 implicate the ceramide pathway in human leukemias

    PubMed Central

    Kim, Woo Jae; Okimoto, Ross A.; Purton, Louise E.; Goodwin, Meagan; Haserlat, Sara M.; Dayyani, Farshid; Sweetser, David A.; McClatchey, Andrea I.; Bernard, Olivier A.; Look, A. Thomas; Bell, Daphne W.; Scadden, David T.

    2008-01-01

    Ceramide is a lipid second messenger derived from the hydrolysis of sphingomyelin by sphingomyelinases (SMases) and implicated in diverse cellular responses, including growth arrest, differentiation, and apoptosis. Defects in the neutral SMase (nSMase) gene Smpd3, the primary regulator of ceramide biosynthesis, are responsible for developmental defects of bone; regulation of ceramide levels have been implicated in macrophage differentiation, but this pathway has not been directly implicated in human cancer. In a genomic screen for gene copy losses contributing to tumorigenesis in a mouse osteosarcoma model, we identified a somatic homozygous deletion specifically targeting Smpd3. Reconstitution of SMPD3 expression in mouse tumor cells lacking the endogenous gene enhanced tumor necrosis factor (TNF)–induced reduction of cell viability. Nucleotide sequencing of the highly conserved SMPD3 gene in a large panel of human cancers revealed mutations in 5 (5%) of 92 acute myeloid leukemias (AMLs) and 8 (6%) of 131 acute lymphoid leukemias (ALLs), but not in other tumor types. In a subset of these mutations, functional analysis indicated defects in protein stability and localization. Taken together, these observations suggest that disruption of the ceramide pathway may contribute to a subset of human leukemias. PMID:18299447

  10. Neutral Sphingomyelinase 2 (nSMase2) Is a Phosphoprotein Regulated by Calcineurin (PP2B)*

    PubMed Central

    Filosto, Simone; Fry, William; Knowlton, Anne A.; Goldkorn, Tzipora

    2010-01-01

    We previously reported that exposure of human airway epithelial cells to oxidative stress increased ceramide generation via specific activation of neutral sphingomyelinase2 (nSMase2). Here we show that nSMase2 is a phosphoprotein exclusively phosphorylated at serine residues. The level of nSMase2 phosphorylation can be modulated by treatment with anisomycin or phorbol 12-myristate 13-acetate (PMA/12-O-tetradecanoylphorbol-13-acetate), suggesting that p38 mitogen-activated protein kinase (MAPK) and protein kinases Cs are upstream of nSMase2 phosphorylation. Oxidative stress enhances both the activity and phosphorylation of nSMase2. Strikingly, we show here that nSMase2 is bound directly by the phosphatase calcineurin (CaN), which acts as an on/off switch for nSMase2 phosphorylation in the presence or absence of oxidative stress. Specifically, CaN is being inhibited/degraded and therefore does not bind nSMase2 under oxidative stress, and a mutant nSMase2 that lacks the CaN binding site exhibits constitutively elevated phosphorylation and increased activity relative to wild type nSMase2. Importantly, the phosphorylation and activity of the mutant no longer responds to oxidative stress, confirming that CaN is the critical link that allows oxidative stress to modulate nSMase2 phosphorylation and function. PMID:20106976

  11. Bidirectional control of sphingomyelinase activity and surface topography in lipid monolayers.

    PubMed Central

    Fanani, María Laura; Härtel, Steffen; Oliveira, Rafael G; Maggio, Bruno

    2002-01-01

    Lipid lateral organization is increasingly found to modulate membrane-bound enzymes. We followed in real time the reaction course of sphingomyelin (SM) degradation by Bacillus cereus sphingomyelinase (SMase) of lipid monolayers by epifluorescence microscopy. There is evidence that formation of ceramide (Cer), a lipid second messenger, drives structural reorganization of membrane lipids. Our results provide visual evidence that SMase activity initially alters surface topography by inducing phase separation into condensed (Cer-enriched) and expanded (SM-enriched) domains. The Cer-enriched phase grows steadily as the reaction proceeds at a constant rate. The surface topography derived from the SMase-driven reaction was compared with, and found to differ from, that of premixed SM/Cer monolayers of the same lipid composition, indicating that substantial information content is stored depending on the manner in which the surface was generated. The long-range topographic changes feed back on the kinetics of Smase, and the onset of condensed-phase percolation is temporally correlated with a rapid drop of reaction rate. These observations reveal a bidirectional influence and communication between effects taking place at the local molecular level and the supramolecular organization. The results suggest a novel biocatalytic-topographic mechanism in which a surface enzymatic activity can influence the function of amphitropic proteins important for cell function. PMID:12496108

  12. Constitutive overexpression of asm18 increases the production and diversity of maytansinoids in Actinosynnema pretiosum.

    PubMed

    Li, Shanren; Lu, Chunhua; Chang, Xiaoyan; Shen, Yuemao

    2016-03-01

    Ansamitocins isolated from Actinosynnema pretiosum, potent antitumor compounds, belong to the family of maytansinoids, and the antibody-maytansinoid conjugates are currently under different phases of clinical trials. The clinical applications of ansamitocins have stimulated extensive studies to improve their production yields. In this study, we investigated the function of a pathway-specific S treptomyces antibiotic regulatory protein (SARP) family regulator, Asm18, and observed that ectopic overexpression of the asm18 gene increased the production of N-demethyl-4,5-desepoxy-maytansinol (2) to 50 mg/L in the HGF052 + pJTU824-asm18 strain, an increase by 4.7-fold compared to that of the control strain HGF052 + pJTU824. Real-time PCR analysis showed that the overexpression of the asm18 gene selectively increased the transcription levels of the genes involved in the biosynthesis of the starter unit (asm43), polyketide assembly (asmA), post-PKS modification (asm21), as well as the transcription levels of the regulatory gene (asm8), which is a specific LAL-type activator in ansamitocin biosynthesis. With the increase of fermentation titre, seven ansamitocin analogs (1-7) including three new ones (1, 5, and 6) and maytansinol (7) were isolated from the HGF052 + pJTU824-asm18 strain. Our results not only pave the way for further improving the production of ansamitocin analogs but also indicate that the post-PKS modifications of ansamitocin biosynthesis are flexible, which brings a potential of producing maytansinol, the most fascinating intermediate for the synthesis of antibody-maytansinoid conjugates, by optimizing the HGF052 and/or HGF052 + pJTU824-asm18 strains. PMID:26572523

  13. Encapsulating connections on SoC designs using ASM++ charts

    NASA Astrophysics Data System (ADS)

    de Pablo, Santiago; Herrero, Luis C.; Martínez, Fernando; Rey, Alexis B.

    This article presents a methodology to encapsulate, not only the functionality of several SoC modules, but also the connections between those modules. To achieve these results, the possibilities of Algorithmic State Machines (ASM charts) have been extended to develop a compiler. Using this approach, a SoC design becomes a set of chart boxes and links: several boxes describe parameterized modules in a hierarchical fashion, other boxes encapsulate their connections, and all boxes are linked together using simple lines. At last, a compiler processes all required files and generates the corresponding VHDL or Verilog code, valid for simulation and synthesis. A small SoC design with two DSP processors is shown as an example.

  14. ASM stepper alignment through thick epitaxial silicon films

    NASA Astrophysics Data System (ADS)

    Black, Iain

    1999-04-01

    High voltage bipolar and BiCMOS processes often use thick epitaxially grown layers of silicon. These films 12-24 micrometers thick offer a considerable challenge to the alignment of subsequent process layers due to the 'wash out' and image distortion, caused to any underlying pattern, which render automatic alignment mark recognition difficult it not impossible. Historically using projection aligner technology these immediately post Epi layers have been manually aligned with future automatic alignment target defined at the first opportunity post Epi. This is not possible using ASM steppers, as these depend upon marks etched into the silicon, before first processing, to create marks, to which all subsequent layers are registered. To allow the stepper to run wafers with these Epi films a new approach was required.

  15. Rapid turn-over of plasma membrane sphingomyelin and cholesterol in baby hamster kidney cells after exposure to sphingomyelinase.

    PubMed

    Slotte, J P; Härmälä, A S; Jansson, C; Pörn, M I

    1990-12-14

    Plasma membrane sphingomyelin in baby hamster kidney (BHK-21) cells was hydrolyzed with sphingomyelinase (Staphylococcus aureus) and the effects on membrane cholesterol translocation and the properties of membrane bound adenylate cyclase and Na+/K(+)-ATPase were determined. Exposure of confluent BHK-21 cells to 0.1 U/ml of sphingomyelinase led to the degradation (at 37 degrees C) of about 60% of cell sphingomyelin. No simultaneous hydrolysis of phosphatidylcholine occurred. The hydrolysis of sphingomyelin subsequently led to the translocation (within 40 min) of about 50-60% of cell [3H]cholesterol from a cholesterol oxidase susceptible pool to an oxidase resistant compartment. The translocation of [3H]cholesterol from the cell surface to intracellular membranes was accompanied by a paralleled increase in [3H]cholesterol ester formation. When cells were first exposed to sphingomyelinase (to degrade sphingomyelin) and then incubated without the enzyme in serum-free media, the mass of cell sphingomyelin decreased initially (by 60%), but then began to increase and reached control levels within 3-4 h. The rapid re-synthesis of sphingomyelin was accompanied by an equally rapid normalization of cell [3H]cholesterol distribution. The re-formation of cell sphingomyelin also led to a decreased content of cellular [3H]cholesterol esters, indicating that unesterified [3H]cholesterol was pulled out of the cholesterol ester cycle and transported to the cell surface. Exposure of BHK-21 cells to sphingomyelinase further led to a dramatically decreased activity of ouabain-sensitive Na+/K(+)-ATPase, whereas forskolin-stimulated adenylate cyclase activity was not affected. The activity of Na+/K(+)-ATPase returned to normal in parallel with the normalization of cell sphingomyelin mass and cholesterol distribution. We conclude that sphingomyelin has profound effects on the steady-state distribution of cell cholesterol, and that manipulations of cell sphingomyelin levels directly and

  16. Neutral sphingomyelinase-3 mediates TNF-stimulated oxidant activity in skeletal muscle

    PubMed Central

    Moylan, Jennifer S.; Smith, Jeffrey D.; Wolf Horrell, Erin M.; McLean, Julie B.; Deevska, Gergana M.; Bonnell, Mark R.; Nikolova-Karakashian, Mariana N.; Reid, Michael B.

    2014-01-01

    Aims Sphingolipid and oxidant signaling affect glucose uptake, atrophy, and force production of skeletal muscle similarly and both are stimulated by tumor necrosis factor (TNF), suggesting a connection between systems. Sphingolipid signaling is initiated by neutral sphingomyelinase (nSMase), a family of agonist-activated effector enzymes. Northern blot analyses suggest that nSMase3 may be a striated muscle-specific nSMase. The present study tested the hypothesis that nSMase3 protein is expressed in skeletal muscle and functions to regulate TNF-stimulated oxidant production. Results We demonstrate constitutive nSMase activity in skeletal muscles of healthy mice and humans and in differentiated C2C12 myotubes. nSMase3 (Smpd4 gene) mRNA is highly expressed in muscle. An nSMase3 protein doublet (88 and 85 kD) is derived from alternative mRNA splicing of exon 11. The proteins partition differently. The full-length 88 kD isoform (nSMase3a) fractionates with membrane proteins that are resistant to detergent extraction; the 85 kD isoform lacking exon 11 (nSMase3b) is more readily extracted and fractionates with detergent soluble membrane proteins; neither variant is detected in the cytosol. By immunofluorescence microscopy, nSMase3 resides in both internal and sarcolemmal membranes. Finally, myotube nSMase activity and cytosolic oxidant activity are stimulated by TNF. Both if these responses are inhibited by nSMase3 knockdown. Innovation These findings identify nSMase3 as an intermediate that links TNF receptor activation, sphingolipid signaling, and skeletal muscle oxidant production. Conclusion Our data show that nSMase3 acts as a signaling nSMase in skeletal muscle that is essential for TNF-stimulated oxidant activity. PMID:25180167

  17. Sphingomyelinase D inhibits store-operated Ca2+ entry in T lymphocytes by suppressing ORAI current

    PubMed Central

    Combs, David J.

    2015-01-01

    Infections caused by certain bacteria including Mycobacterium tuberculosis and Corynebacterium pseudotuberculosis provoke inflammatory responses characterized by the formation of granulomas with necrotic foci—so-called caseous necrosis. The granulomas of infected animals show prominent infiltration by T lymphocytes, and T cell depletion increases host mortality. Notorious zoonotic C. pseudotuberculosis secretes sphingomyelinase (SMase) D, a phospholipase that cleaves off the choline moiety of sphingomyelin, a phospholipid found primarily in the outer leaflet of host cell plasma membranes. Experimental C. pseudotuberculosis strains that lack SMase D are markedly less infectious and unable to spread in hosts, indicating that this enzyme is a crucial virulence factor for sustaining the caseous lymphadenitis infections caused by this microbe. However, the molecular mechanism by which SMase D helps bacteria evade the host’s immune response remains unknown. Here, we find that SMase D inhibits store-operated Ca2+ entry (SOCE) in human T cells and lowers the production of the SOCE-dependent cytokines interleukin-2, which is critical for T cell growth, proliferation, and differentiation, and tumor necrosis factor α, which is crucial for the formation and maintenance of granulomas in microbial infections. SMase D inhibits SOCE through a previously unknown mechanism, namely, suppression of Orai1 current, rather than through altering gating of voltage-gated K+ channels. This finding suggests that, whereas certain genetic mutations abolish Orai1 activity causing severe combined immunodeficiency (SCID), bacteria have the ability to suppress Orai1 activity with SMase D to create an acquired, chronic SCID-like condition that allows persistent infection. Thus, in an example of how virulence factors can disrupt key membrane protein function by targeting phospholipids in host cell membranes, our study has uncovered a novel molecular mechanism that bacteria can use to thwart host

  18. Phosphatidylcholine-Specific Phospholipase C and Sphingomyelinase Activities in Bacteria of the Bacillus cereus Group

    PubMed Central

    Pomerantsev, A. P.; Kalnin, K. V.; Osorio, M.; Leppla, S. H.

    2003-01-01

    Bacillus anthracis is nonhemolytic, even though it is closely related to the highly hemolytic Bacillus cereus. Hemolysis by B. cereus results largely from the action of phosphatidylcholine-specific phospholipase C (PC-PLC) and sphingomyelinase (SPH), encoded by the plc and sph genes, respectively. In B. cereus, these genes are organized in an operon regulated by the global regulator PlcR. B. anthracis contains a highly similar cereolysin operon, but it is transcriptionally silent because the B. anthracis PlcR is truncated at the C terminus. Here we report the cloning, expression, purification, and enzymatic characterization of PC-PLC and SPH from B. cereus and B. anthracis. We also investigated the effects of expressing PlcR on the expression of plc and sph. In B. cereus, PlcR was found to be a positive regulator of plc but a negative regulator of sph. Replacement of the B. cereus plcR gene by its truncated orthologue from B. anthracis eliminated the activities of both PC-PLC and SPH, whereas introduction into B. anthracis of the B. cereus plcR gene with its own promoter did not activate cereolysin expression. Hemolytic activity was detected in B. anthracis strains containing the B. cereus plcR gene on a multicopy plasmid under control of the strong B. anthracis protective antigen gene promoter or in a strain carrying a multicopy plasmid containing the entire B. cereus plc-sph operon. Slight hemolysis and PC-PLC activation were found when PlcR-producing B. anthracis strains were grown under anaerobic-plus-CO2 or especially under aerobic-plus-CO2 conditions. Unmodified parental B. anthracis strains did not demonstrate obvious hemolysis under the same conditions. PMID:14573681

  19. The First ASME Code Stamped Cryomodule at SNS

    SciTech Connect

    Howell, M P; Crofford, M T; Douglas, D L; Kim, S -H; Steward, S T; Strong, W H; Afanador, R; Hannah, B S; Saunders, J; Mammosser, J D

    2012-07-01

    The first spare cryomodule for the Spallation Neutron Source (SNS) has been designed, fabricated, and tested by SNS personnel. The approach to design for this cryomodule was to hold critical design features identical to the original design such as bayonet positions, coupler positions, cold mass assembly, and overall footprint. However, this is the first SNS cryomodule that meets the pressure requirements put forth in the 10 CFR 851: Worker Safety and Health Program. The most significant difference is that Section VIII of the ASME Boiler and Pressure Vessel Code was applied to the vacuum vessel of this cryomodule. Applying the pressure code to the helium vessels within the cryomodule was considered. However, it was determined to be schedule prohibitive because it required a code case for materials that are not currently covered by the code. Good engineering practice was applied to the internal components to verify the quality and integrity of the entire cryomodule. The design of the cryomodule, fabrication effort, and cryogenic test results will be reported in this paper.

  20. Estimates of margins in ASME Code strength values for stainless steel nuclear piping

    SciTech Connect

    Ware, A.G.

    1995-11-01

    The margins in the ASME Code stainless steel allowable stress values that can be attributed to the variations in material strength are evaluated for nuclear piping steels. Best-fit curves were calculated for the material test data that were used to determine allowable stress values for stainless steels in the ASME Code, supplemented by more recent data, to estimate the mean stresses. The mean yield stresses (on which the stainless steel S{sub m} values are based) from the test data are about 15 to 20% greater than the ASME Code yield stress values. The ASME Code yield stress values are estimated to approximately coincide with the 97% confidence limit from the test data. The mean and 97% confidence limit values can be used in the probabilistic risk assessments of nuclear piping.

  1. 14 CFR 330.31 - What data must air carriers submit concerning ASMs or RTMs?

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... all-cargo carrier, you must have submitted your RTM reports to the Department for the second calendar... correct an error that you document to the Department, you must not alter the ASM or RTM reports...

  2. 115-year-old society knows how to reach young scientists: ASM Young Ambassador Program.

    PubMed

    Karczewska-Golec, Joanna

    2015-12-25

    With around 40,000 members in more than 150 countries, American Society for Microbiology (ASM) faces the challenge of meeting very diverse needs of its increasingly international members base. The newly launched ASM Young Ambassador Program seeks to aid the Society in this effort. Equipped with ASM conceptual support and financing, Young Ambassadors (YAs) design and pursue country-tailored approaches to strengthen the Society's ties with local microbiological communities. In a trans-national setting, the active presence of YAs at important scientific events, such as 16th European Congress on Biotechnology, forges new interactions between ASM and sister societies. The paper presents an overview of the Young Ambassadors-driven initiatives at both global and country levels, and explores the topic of how early-career scientists can contribute to science diplomacy and international relations. PMID:25449541

  3. 46 CFR 57.02-2 - Adoption of section IX of the ASME Code.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 2 2014-10-01 2014-10-01 false Adoption of section IX of the ASME Code. 57.02-2 Section 57.02-2 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) MARINE ENGINEERING WELDING AND BRAZING General Requirements § 57.02-2 Adoption of section IX of the ASME Code. (a) The qualifications for all types of welders and brazers,...

  4. 46 CFR 57.02-2 - Adoption of section IX of the ASME Code.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 2 2011-10-01 2011-10-01 false Adoption of section IX of the ASME Code. 57.02-2 Section... AND BRAZING General Requirements § 57.02-2 Adoption of section IX of the ASME Code. (a) The... in this part. Table 57.02-1(a)—Limitations and Modifications to the Adoption of section IX of...

  5. Fatigue evaluation of ASME Class 1 components considering the environmental effects

    SciTech Connect

    Neto, M.M.; Maneschy, E.

    1995-12-01

    A discussion considering fatigue design basis (FDB) and fatigue operating basis (FOB) approaches is presented. These two concepts are applied to evaluate the lifetime of typical ASME 3 class 1 components through simplified and detailed stress analysis. The cumulative usage factor (CUF) calculated using S-N fatigue curves available in the ASME 3 are compared to those obtained by S-N curves modified by the reactor environment. Some recommendations are presented to assess the fatigue in nuclear power plants structures.

  6. Activity of sphingomyelinase in rat liver in acute and chronic toxic hepatitis: proportion between peroxidative and phospholipase pathways of lipid bilayer modification.

    PubMed

    Serebrov, V Yu; Kuzmenko, D I; Burov, P G; Novitsky, S V

    2009-01-01

    We showed that sphingomyelinase activity in the liver increased only during the acute phase of toxic hepatitis. Peroxidative modification of hepatocyte membrane bilayer prevailed during the acute phase, while after transformation of the process to the chronic phase phospholipase pathway predominated. PMID:19526125

  7. A Minimal Path Searching Approach for Active Shape Model (ASM)-based Segmentation of the Lung.

    PubMed

    Guo, Shengwen; Fei, Baowei

    2009-03-27

    We are developing a minimal path searching method for active shape model (ASM)-based segmentation for detection of lung boundaries on digital radiographs. With the conventional ASM method, the position and shape parameters of the model points are iteratively refined and the target points are updated by the least Mahalanobis distance criterion. We propose an improved searching strategy that extends the searching points in a fan-shape region instead of along the normal direction. A minimal path (MP) deformable model is applied to drive the searching procedure. A statistical shape prior model is incorporated into the segmentation. In order to keep the smoothness of the shape, a smooth constraint is employed to the deformable model. To quantitatively assess the ASM-MP segmentation, we compare the automatic segmentation with manual segmentation for 72 lung digitized radiographs. The distance error between the ASM-MP and manual segmentation is 1.75 ± 0.33 pixels, while the error is 1.99 ± 0.45 pixels for the ASM. Our results demonstrate that our ASM-MP method can accurately segment the lung on digital radiographs. PMID:24386531

  8. A minimal path searching approach for active shape model (ASM)-based segmentation of the lung

    NASA Astrophysics Data System (ADS)

    Guo, Shengwen; Fei, Baowei

    2009-02-01

    We are developing a minimal path searching method for active shape model (ASM)-based segmentation for detection of lung boundaries on digital radiographs. With the conventional ASM method, the position and shape parameters of the model points are iteratively refined and the target points are updated by the least Mahalanobis distance criterion. We propose an improved searching strategy that extends the searching points in a fan-shape region instead of along the normal direction. A minimal path (MP) deformable model is applied to drive the searching procedure. A statistical shape prior model is incorporated into the segmentation. In order to keep the smoothness of the shape, a smooth constraint is employed to the deformable model. To quantitatively assess the ASM-MP segmentation, we compare the automatic segmentation with manual segmentation for 72 lung digitized radiographs. The distance error between the ASM-MP and manual segmentation is 1.75 +/- 0.33 pixels, while the error is 1.99 +/- 0.45 pixels for the ASM. Our results demonstrate that our ASM-MP method can accurately segment the lung on digital radiographs.

  9. Shape Transitions and Lattice Structuring of Ceramide-Enriched Domains Generated by Sphingomyelinase in Lipid Monolayers

    PubMed Central

    Härtel, Steffen; Fanani, María Laura; Maggio, Bruno

    2005-01-01

    Sphingomyelinases (SMases) hydrolyze the membrane constituent sphingomyelin (SM) to phosphocholine and ceramide (Cer). Growing evidence supports that SMase-induced SM→Cer conversion leads to the formation of lateral Cer-enriched domains which drive structural reorganization in lipid membranes. We previously provided visual evidence in real-time for the formation of Cer-enriched domains in SM monolayers through the action of the neutral Bacillus cereus SMase. In this work, we disclose a succession of discrete morphologic transitions and lateral organization of Cer-enriched domains that underlay the SMase-generated surface topography. We further reveal how these structural parameters couple to the generation of two-dimensional electrostatic fields, based upon the specific orientation of the lipid dipole moments in the Cer-enriched domains. Advanced image processing routines in combination with time-resolved epifluorescence microscopy on Langmuir monolayers revealed: 1), spontaneous nucleation and circular growth of Cer-enriched domains after injection of SMase into the subphase of the SM monolayer; 2), domain-intrinsic discrete transitions from circular to periodically undulating shapes followed by a second transition toward increasingly branched morphologies; 3), lateral superstructure organization into predominantly hexagonal domain lattices; 4), formation of super-superstructures by the hexagonal lattices; and 5), rotationally and laterally coupled domain movement before domain border contact. All patterns proved to be specific for the SMase-driven system since they could not be observed with Cer-enriched domains generated by defined mixtures of SM/Cer in enzyme-free monolayers at the same surface pressure (Π = 10 mN/m). Following the theories of lateral shape transitions, dipolar electrostatic interactions of lipid domains, and direct determinations of the monolayer dipole potential, our data show that SMase induces a domain-specific packing and orientation of

  10. Tuning voltage-gated channel activity and cellular excitability with a sphingomyelinase

    PubMed Central

    Combs, David J.; Shin, Hyeon-Gyu; Xu, Yanping; Ramu, Yajamana

    2013-01-01

    Voltage-gated ion channels generate action potentials in excitable cells and help set the resting membrane potential in nonexcitable cells like lymphocytes. It has been difficult to investigate what kinds of phospholipids interact with these membrane proteins in their native environments and what functional impacts such interactions create. This problem might be circumvented if we could modify specific lipid types in situ. Using certain voltage-gated K+ (KV) channels heterologously expressed in Xenopus laevis oocytes as a model, our group has shown previously that sphingomyelinase (SMase) D may serve this purpose. SMase D is known to remove the choline group from sphingomyelin, a phospholipid primarily present in the outer leaflet of plasma membranes. This SMase D action lowers the energy required for voltage sensors of a KV channel to enter the activated state, causing a hyperpolarizing shift of the Q-V and G-V curves and thus activating them at more hyperpolarized potentials. Here, we find that this SMase D effect vanishes after removing most of the voltage-sensor paddle sequence, a finding supporting the notion that SMase D modification of sphingomyelin molecules alters these lipids’ interactions with voltage sensors. Then, using SMase D to probe lipid–channel interactions, we find that SMase D not only similarly stimulates voltage-gated Na+ (NaV) and Ca2+ channels but also markedly slows NaV channel inactivation. However, the latter effect is not observed in tested mammalian cells, an observation highlighting the profound impact of the membrane environment on channel function. Finally, we directly demonstrate that SMase D stimulates both native KV1.3 in nonexcitable human T lymphocytes at their typical resting membrane potential and native NaV channels in excitable cells, such that it shifts the action potential threshold in the hyperpolarized direction. These proof-of-concept studies illustrate that the voltage-gated channel activity in both excitable and

  11. Risk-based inspection in ASME Section XI

    SciTech Connect

    Lance, J.J.

    1996-12-01

    By 1970 the first edition of the ASME Code Section XI, Inservice Inspection of Nuclear Reactor Coolant Systems was published. From its inception, the Section XI inservice inspection scope was based on a fundamental risk-based selection process. In other words the inservice inspection scope included components where the consequences of a pressure boundary failure were high. Once the consequence significant system boundaries were established, inspections would then be performed at locations believed to be most susceptible service induced failure. Current Section XI requirements require that inspection locations be selected on the basis of peak stress and fatigue usage values contained in the Design Reports. These original stress calculations were designed to qualify a design and assure that the plant would provide reliable service throughout its design life. For the most part, the fatigue usage values in these reports do not provide an accurate measure of service life. As service history has demonstrated, the use of Design Report stresses and fatigue usage values can be misleading. The Section XI ISI inspection requirements have always been intended to focus inspections at those locations in the plant that pose the greater risk to reactor safety. This fundamental principle behind the Section XI inspection requirements has guided Section XI since its inception. However, today Utility resources are limited. The move in many states to deregulate utilities and growing competition from independent power producers is challenging Owners to reduce operating and maintenance cost without sacrificing safety. These programs should allow plants to focus limited resources on those locations where damage mechanisms are active and consequences are high. This will provide for efficient use of plants resources and improve safety.

  12. ASME code considerations for the compact heat exchanger

    SciTech Connect

    Nestell, James; Sham, Sam

    2015-08-31

    . Classic shell and tube designs will be large and costly, and may only be appropriate in steam generator service in the SHX where boiling inside the tubes occurs. For other energy conversion systems, all of these features can be met in a compact heat exchanger design. This report will examine some of the ASME Code issues that will need to be addressed to allow use of a Code-qualified compact heat exchanger in IHX or SHX nuclear service. Most effort will focus on the IHX, since the safety-related (Class A) design rules are more extensive than those for important-to-safety (Class B) or commercial rules that are relevant to the SHX.

  13. Microbial sphingomyelinase induces RhoA-mediated reorganization of the apical brush border membrane and is protective against invasion.

    PubMed

    Saslowsky, David E; Thiagarajah, Jay R; McCormick, Beth A; Lee, Jean C; Lencer, Wayne I

    2016-04-01

    The apical brush border membrane (BBM) of intestinal epithelial cells forms a highly structured and dynamic environmental interface that serves to regulate cellular physiology and block invasion by intestinal microbes and their products. How the BBM dynamically responds to pathogenic and commensal bacterial signals can define intestinal homeostasis and immune function. We previously found that in model intestinal epithelium, the conversion of apical membrane sphingomyelin to ceramide by exogenous bacterial sphingomyelinase (SMase) protected against the endocytosis and toxicity of cholera toxin. Here we elucidate a mechanism of action by showing that SMase induces a dramatic, reversible, RhoA-dependent alteration of the apical cortical F-actin network. Accumulation of apical membrane ceramide is necessary and sufficient to induce the actin phenotype, and this coincides with altered membrane structure and augmented innate immune function as evidenced by resistance to invasion by Salmonella. PMID:26864627

  14. Microbial sphingomyelinase induces RhoA-mediated reorganization of the apical brush border membrane and is protective against invasion

    PubMed Central

    Saslowsky, David E.; Thiagarajah, Jay R.; McCormick, Beth A.; Lee, Jean C.; Lencer, Wayne I.

    2016-01-01

    The apical brush border membrane (BBM) of intestinal epithelial cells forms a highly structured and dynamic environmental interface that serves to regulate cellular physiology and block invasion by intestinal microbes and their products. How the BBM dynamically responds to pathogenic and commensal bacterial signals can define intestinal homeostasis and immune function. We previously found that in model intestinal epithelium, the conversion of apical membrane sphingomyelin to ceramide by exogenous bacterial sphingomyelinase (SMase) protected against the endocytosis and toxicity of cholera toxin. Here we elucidate a mechanism of action by showing that SMase induces a dramatic, reversible, RhoA-dependent alteration of the apical cortical F-actin network. Accumulation of apical membrane ceramide is necessary and sufficient to induce the actin phenotype, and this coincides with altered membrane structure and augmented innate immune function as evidenced by resistance to invasion by Salmonella. PMID:26864627

  15. Inhibition of neutral sphingomyelinase decreases elevated levels of inducible nitric oxide synthase and apoptotic cell death in ocular hypertensive rats

    SciTech Connect

    Aslan, Mutay; Basaranlar, Goksun; Unal, Mustafa; Ciftcioglu, Akif; Derin, Narin; Mutus, Bulent

    2014-11-01

    Endoplasmic reticulum (ER) stress and excessive nitric oxide production via induction of inducible nitric oxide synthase (NOS2) have been implicated in the pathogenesis of neuronal retinal cell death in ocular hypertension. Neutral sphingomyelinase (N-SMase)/ceramide pathway can regulate NOS2 expression, hence this study determined the role of selective neutral sphingomyelinase (N-SMase) inhibition on retinal NOS2 levels, ER stress, apoptosis and visual evoked potentials (VEPs) in a rat model of elevated intraocular pressure (EIOP). NOS2 expression and retinal protein nitration were significantly greater in EIOP and significantly decreased with N-SMase inhibition. A significant increase was observed in retinal ER stress markers pPERK, CHOP and GRP78 in EIOP, which were not significantly altered by N-SMase inhibition. Retinal TUNEL staining showed increased apoptosis in all EIOP groups; however N-SMase inhibition significantly decreased the percent of apoptotic cells in EIOP. Caspase-3, -8 and -9 activities were significantly increased in EIOP and returned to baseline levels following N-SMase inhibition. Latencies of all VEP components were significantly prolonged in EIOP and shortened following N-SMase inhibition. Data confirm the role of nitrative injury in EIOP and highlight the protective effect of N-SMase inhibition in EIOP via down-regulation of NOS2 levels and nitrative stress. - Highlights: • Inhibition of N-SMase decreases NOS2 levels in ocular hypertension. • Inhibition of N-SMase decreases protein nitration in ocular hypertension. • Inhibition of N-SMase decreases caspase activation in ocular hypertension. • Inhibition of N-SMase decreases apoptosis in ocular hypertension.

  16. An ASM/ADM model interface for dynamic plant-wide simulation.

    PubMed

    Nopens, Ingmar; Batstone, Damien J; Copp, John B; Jeppsson, Ulf; Volcke, Eveline; Alex, Jens; Vanrolleghem, Peter A

    2009-04-01

    Mathematical modelling has proven to be very useful in process design, operation and optimisation. A recent trend in WWTP modelling is to include the different subunits in so-called plant-wide models rather than focusing on parts of the entire process. One example of a typical plant-wide model is the coupling of an upstream activated sludge plant (including primary settler, and secondary clarifier) to an anaerobic digester for sludge digestion. One of the key challenges when coupling these processes has been the definition of an interface between the well accepted activated sludge model (ASM1) and anaerobic digestion model (ADM1). Current characterisation and interface models have key limitations, the most critical of which is the over-use of X(c) (or lumped complex) variable as a main input to the ADM1. Over-use of X(c) does not allow for variation of degradability, carbon oxidation state or nitrogen content. In addition, achieving a target influent pH through the proper definition of the ionic system can be difficult. In this paper, we define an interface and characterisation model that maps degradable components directly to carbohydrates, proteins and lipids (and their soluble analogues), as well as organic acids, rather than using X(c). While this interface has been designed for use with the Benchmark Simulation Model No. 2 (BSM2), it is widely applicable to ADM1 input characterisation in general. We have demonstrated the model both hypothetically (BSM2), and practically on a full-scale anaerobic digester treating sewage sludge. PMID:19232670

  17. The ASM Curriculum Guidelines for Undergraduate Microbiology: A Case Study of the Advocacy Role of Societies in Reform Efforts

    PubMed Central

    Horak, Rachel E. A.; Merkel, Susan; Chang, Amy

    2015-01-01

    A number of national reports, including Vision and Change in Undergraduate Biology Education: A Call to Action, have called for drastic changes in how undergraduate biology is taught. To that end, the American Society for Microbiology (ASM) has developed new Curriculum Guidelines for undergraduate microbiology that outline a comprehensive curriculum for any undergraduate introductory microbiology course or program of study. Designed to foster enduring understanding of core microbiology concepts, the Guidelines work synergistically with backwards course design to focus teaching on student-centered goals and priorities. In order to qualitatively assess how the ASM Curriculum Guidelines are used by educators and learn more about the needs of microbiology educators, the ASM Education Board distributed two surveys to the ASM education community. In this report, we discuss the results of these surveys (353 responses). We found that the ASM Curriculum Guidelines are being implemented in many different types of courses at all undergraduate levels. Educators indicated that the ASM Curriculum Guidelines were very helpful when planning courses and assessments. We discuss some specific ways in which the ASM Curriculum Guidelines have been used in undergraduate classrooms. The survey identified some barriers that microbiology educators faced when trying to adopt the ASM Curriculum Guidelines, including lack of time, lack of financial resources, and lack of supporting resources. Given the self-reported challenges to implementing the ASM Curriculum Guidelines in undergraduate classrooms, we identify here some activities related to the ASM Curriculum Guidelines that the ASM Education Board has initiated to assist educators in the implementation process. PMID:25949769

  18. Swarm's Absolute Magnetometer (ASM) Experimental Vector Mode, a Unique Capability With Considerable Potential

    NASA Astrophysics Data System (ADS)

    Hulot, G.; Vigneron, P.; Leger, J. M.; Fratter, I.; Olsen, N.; Jager, T.; Bertrand, F.; Brocco, L.; Sirol, O.; Lalanne, X.; Boness, A.; Cattin, V.

    2014-12-01

    In addition to providing the reference absolute scalar measurements on the three Alpha, Bravo and Charlie satellites of the Swarm mission, each Absolute Scalar Magnetometer (ASM) can also, and simultaneously, provide independent experimental vector field measurements. These experimental data appear to be of the expected quality (though somewhat less so on Charlie). In addition, comparisons of these measurements with readings from the Vector Field Magnetometer (VFM, located some distance away along the boom and used to produce the nominal Swarm Level1b vector data) show that the mechanical link between both instruments is very stable on all three satellites. These remarkable circumstances make it possible not only to compare the output of the VFM and ASM instruments for cross-validation purposes, but also to compute geomagnetic field models using only ASM scalar and vector mode data, without resorting to VFM data. Such models can then be compared to models computed in exactly the same way from VFM data (using exactly the same data distribution in time and space for both models, which thus only differ by the fact that the data are provided by either the ASM experimental vector mode, or the nominal L1b calibrated VFM data). As we shall illustrate in this presentation, such comparisons provide extremely valuable information. Not only do they show that ASM experimental vector mode data can indeed be used to produce high-degree geomagnetic field models, but they also provide a very interesting perspective on what may be the cause of undesired perturbations on either of the ASM and VFM instruments.

  19. 46 CFR 56.01-5 - Adoption of ASME B31.1 for power piping, and other standards.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... accordance with ASME B31.1 (incorporated by reference; see 46 CFR 56.01-2), as limited, modified, or replaced... subchapter. See 46 CFR 56.60-1(b) for the other adopted commercial standards applicable to piping systems... 46 Shipping 2 2013-10-01 2013-10-01 false Adoption of ASME B31.1 for power piping, and...

  20. 46 CFR 56.01-5 - Adoption of ASME B31.1 for power piping, and other standards.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... accordance with ASME B31.1 (incorporated by reference; see 46 CFR 56.01-2), as limited, modified, or replaced... subchapter. See 46 CFR 56.60-1(b) for the other adopted commercial standards applicable to piping systems... 46 Shipping 2 2011-10-01 2011-10-01 false Adoption of ASME B31.1 for power piping, and...

  1. 46 CFR 56.01-5 - Adoption of ASME B31.1 for power piping, and other standards.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... accordance with ASME B31.1 (incorporated by reference; see 46 CFR 56.01-2), as limited, modified, or replaced... subchapter. See 46 CFR 56.60-1(b) for the other adopted commercial standards applicable to piping systems... 46 Shipping 2 2014-10-01 2014-10-01 false Adoption of ASME B31.1 for power piping, and...

  2. 46 CFR 56.01-5 - Adoption of ASME B31.1 for power piping, and other standards.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... accordance with ASME B31.1 (incorporated by reference; see 46 CFR 56.01-2), as limited, modified, or replaced... subchapter. See 46 CFR 56.60-1(b) for the other adopted commercial standards applicable to piping systems... 46 Shipping 2 2012-10-01 2012-10-01 false Adoption of ASME B31.1 for power piping, and...

  3. 46 CFR 56.01-5 - Adoption of ASME B31.1 for power piping, and other standards.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... accordance with ASME B31.1 (incorporated by reference; see 46 CFR 56.01-2), as limited, modified, or replaced... subchapter. See 46 CFR 56.60-1(b) for the other adopted commercial standards applicable to piping systems... 46 Shipping 2 2010-10-01 2010-10-01 false Adoption of ASME B31.1 for power piping, and...

  4. 46 CFR 52.01-2 - Adoption of section I of the ASME Boiler and Pressure Vessel Code.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... (incorporated by reference; see 46 CFR 52.01-1), as limited, modified, or replaced by specific requirements in... 46 Shipping 2 2010-10-01 2010-10-01 false Adoption of section I of the ASME Boiler and Pressure...) MARINE ENGINEERING POWER BOILERS General Requirements § 52.01-2 Adoption of section I of the ASME...

  5. 46 CFR 52.01-2 - Adoption of section I of the ASME Boiler and Pressure Vessel Code.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... (incorporated by reference; see 46 CFR 52.01-1), as limited, modified, or replaced by specific requirements in... 46 Shipping 2 2014-10-01 2014-10-01 false Adoption of section I of the ASME Boiler and Pressure...) MARINE ENGINEERING POWER BOILERS General Requirements § 52.01-2 Adoption of section I of the ASME...

  6. 46 CFR 52.01-2 - Adoption of section I of the ASME Boiler and Pressure Vessel Code.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... (incorporated by reference; see 46 CFR 52.01-1), as limited, modified, or replaced by specific requirements in... 46 Shipping 2 2013-10-01 2013-10-01 false Adoption of section I of the ASME Boiler and Pressure...) MARINE ENGINEERING POWER BOILERS General Requirements § 52.01-2 Adoption of section I of the ASME...

  7. 46 CFR 52.01-2 - Adoption of section I of the ASME Boiler and Pressure Vessel Code.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... (incorporated by reference; see 46 CFR 52.01-1), as limited, modified, or replaced by specific requirements in... 46 Shipping 2 2011-10-01 2011-10-01 false Adoption of section I of the ASME Boiler and Pressure...) MARINE ENGINEERING POWER BOILERS General Requirements § 52.01-2 Adoption of section I of the ASME...

  8. 46 CFR 52.01-2 - Adoption of section I of the ASME Boiler and Pressure Vessel Code.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... (incorporated by reference; see 46 CFR 52.01-1), as limited, modified, or replaced by specific requirements in... 46 Shipping 2 2012-10-01 2012-10-01 false Adoption of section I of the ASME Boiler and Pressure...) MARINE ENGINEERING POWER BOILERS General Requirements § 52.01-2 Adoption of section I of the ASME...

  9. Simulation and optimization of a coking wastewater biological treatment process by activated sludge models (ASM).

    PubMed

    Wu, Xiaohui; Yang, Yang; Wu, Gaoming; Mao, Juan; Zhou, Tao

    2016-01-01

    Applications of activated sludge models (ASM) in simulating industrial biological wastewater treatment plants (WWTPs) are still difficult due to refractory and complex components in influents as well as diversity in activated sludges. In this study, an ASM3 modeling study was conducted to simulate and optimize a practical coking wastewater treatment plant (CWTP). First, respirometric characterizations of the coking wastewater and CWTP biomasses were conducted to determine the specific kinetic and stoichiometric model parameters for the consecutive aeration-anoxic-aeration (O-A/O) biological process. All ASM3 parameters have been further estimated and calibrated, through cross validation by the model dynamic simulation procedure. Consequently, an ASM3 model was successfully established to accurately simulate the CWTP performances in removing COD and NH4-N. An optimized CWTP operation condition could be proposed reducing the operation cost from 6.2 to 5.5 €/m(3) wastewater. This study is expected to provide a useful reference for mathematic simulations of practical industrial WWTPs. PMID:26439861

  10. Evaluation of ASME Section XI Reference Level Sensitivity for Initiation of Ultrasonic Inspection Examination

    SciTech Connect

    Taylor, T. T.; Selby, G. P.

    1981-04-01

    This report evaluates the change in inspection sensitivity resulting in major changes of ASME Boiler and Pressure Vessel Code Section XI between 1974 and 1977 Editions. It was found that the inspection sensitivity resulting from requirements of the 1977 Edition of Section XI were not adequate to detect minimum flaws referenced by same Code.

  11. PHASE I MATERIALS PROPERTY DATABASE DEVELOPMENT FOR ASME CODES AND STANDARDS

    SciTech Connect

    Ren, Weiju; Lin, Lianshan

    2013-01-01

    To support the ASME Boiler and Pressure Vessel Codes and Standard (BPVC) in modern information era, development of a web-based materials property database is initiated under the supervision of ASME Committee on Materials. To achieve efficiency, the project heavily draws upon experience from development of the Gen IV Materials Handbook and the Nuclear System Materials Handbook. The effort is divided into two phases. Phase I is planned to deliver a materials data file warehouse that offers a depository for various files containing raw data and background information, and Phase II will provide a relational digital database that provides advanced features facilitating digital data processing and management. Population of the database will start with materials property data for nuclear applications and expand to data covering the entire ASME Code and Standards including the piping codes as the database structure is continuously optimized. The ultimate goal of the effort is to establish a sound cyber infrastructure that support ASME Codes and Standards development and maintenance.

  12. Status of ASME Section III Task Group on Graphite Support Core Structures

    SciTech Connect

    Robert L. Bratton; Tim D. Burchell

    2005-08-01

    This report outlines the roadmap that the ASME Project Team on Graphite Core Supports is pursuing to establish design codes for unirradiated and irradiated graphite core components during its first year of operation. It discusses the deficiencies in the proposed Section III, Division 2, Subsection CE graphite design code and the different approaches the Project Team has taken to address those deficiencies.

  13. 14 CFR 330.31 - What data must air carriers submit concerning ASMs or RTMs?

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 4 2012-01-01 2012-01-01 false What data must air carriers submit concerning ASMs or RTMs? 330.31 Section 330.31 Aeronautics and Space OFFICE OF THE SECRETARY, DEPARTMENT OF TRANSPORTATION (AVIATION PROCEEDINGS) PROCEDURAL REGULATIONS PROCEDURES FOR COMPENSATION OF AIR...

  14. 14 CFR 330.31 - What data must air carriers submit concerning ASMs or RTMs?

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 4 2011-01-01 2011-01-01 false What data must air carriers submit concerning ASMs or RTMs? 330.31 Section 330.31 Aeronautics and Space OFFICE OF THE SECRETARY, DEPARTMENT OF TRANSPORTATION (AVIATION PROCEEDINGS) PROCEDURAL REGULATIONS PROCEDURES FOR COMPENSATION OF AIR...

  15. 14 CFR 330.31 - What data must air carriers submit concerning ASMs or RTMs?

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 14 Aeronautics and Space 4 2014-01-01 2014-01-01 false What data must air carriers submit concerning ASMs or RTMs? 330.31 Section 330.31 Aeronautics and Space OFFICE OF THE SECRETARY, DEPARTMENT OF TRANSPORTATION (AVIATION PROCEEDINGS) PROCEDURAL REGULATIONS PROCEDURES FOR COMPENSATION OF AIR...

  16. 14 CFR 330.31 - What data must air carriers submit concerning ASMs or RTMs?

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 14 Aeronautics and Space 4 2013-01-01 2013-01-01 false What data must air carriers submit concerning ASMs or RTMs? 330.31 Section 330.31 Aeronautics and Space OFFICE OF THE SECRETARY, DEPARTMENT OF TRANSPORTATION (AVIATION PROCEEDINGS) PROCEDURAL REGULATIONS PROCEDURES FOR COMPENSATION OF AIR...

  17. Interpreting ASME limits and philosophy in FEA of pressure vessel parts

    SciTech Connect

    Bezerra, L.M.; Cruz, J.R.B.; Miranda, C.A.J.; Neto, M.M.

    1995-12-01

    In recent years there has been an effort to interpret finite element (FE) stress results on the light of the ASME B and PV rules and philosophy. Many task groups have issued guidelines on stress linearization and classifications. All those attempts have come up trying to cope modern FE techniques with the rules imposed by the ASME Code. This paper is an independent contribution to the Pressure Vessel Research Council (PVRC) groups which are studying the stress classification and the failure mechanism in a FE framework. This work tries to complement the interesting work by Hollinger and Hechmer presented in the PVP-94 in Minneapolis. In that paper, the authors examined a typical support skirt and showed relations between the skirt collapse load obtained by finite element analysis and the loads allowed from the ASME stress limits. To complement such paper, in the present article, different skirt geometry configurations are analyzed. The configurations here investigated consist of similar support skirts but with different angles of attachments between cylinder and cone parts. It will be possible to observe the influence of the bending stress in the collapse load and its relation to the allowable loads inferred from the ASME limits. A pressure vessel with torispherical head under internal pressure is also examined. Using elastic and limit load FEA, the present paper determines the collapse loads of the configurations. It sets up the relations between these collapse loads, stress categories, and limits dictated by the ASME Code Subsection NB. On the light of NB rules and philosophy, this paper shows how different methods of stress assessment, classification, and limits may influence in the design of a pressure vessel.

  18. Venom of the Brazilian Spider Sicarius ornatus (Araneae, Sicariidae) Contains Active Sphingomyelinase D: Potential for Toxicity after Envenomation

    PubMed Central

    Lopes, Priscila Hess; Bertani, Rogério; Gonçalves-de-Andrade, Rute M.; Nagahama, Roberto H.; van den Berg, Carmen W.; Tambourgi, Denise V.

    2013-01-01

    Background The spider family Sicariidae includes two genera, Sicarius and Loxosceles. Bites by Sicarius are uncommon in humans and, in Brazil, a single report is known of a 17-year old man bitten by a Sicarius species that developed a necrotic lesion similar to that caused by Loxosceles. Envenomation by Loxosceles spiders can result in dermonecrosis and severe ulceration. Sicarius and Loxosceles spider venoms share a common characteristic, i.e., the presence of Sphingomyelinases D (SMase D). We have previously shown that Loxosceles SMase D is the enzyme responsible for the main pathological effects of the venom. Recently, it was demonstrated that Sicarius species from Africa, like Loxosceles spiders from the Americas, present high venom SMase D activity. However, despite the presence of SMase D like proteins in venoms of several New World Sicarius species, they had reduced or no detectable SMase D activity. In order to contribute to a better understanding about the toxicity of New World Sicarius venoms, the aim of this study was to characterize the toxic properties of male and female venoms from the Brazilian Sicarius ornatus spider and compare these with venoms from Loxosceles species of medical importance in Brazil. Methodology/Principal Findings SDS-PAGE analysis showed variations in the composition of Loxosceles spp. and Sicarius ornatus venoms. Differences in the electrophoretic profiles of male and female venoms were also observed, indicating a possible intraspecific variation in the composition of the venom of Sicarius spider. The major component in all tested venoms had a Mr of 32–35 kDa, which was recognized by antiserum raised against Loxosceles SMases D. Moreover, male and female Sicarius ornatus spiders' venoms were able to hydrolyze sphingomyelin, thus showing an enzymatic activity similar to that determined for Loxosceles venoms. Sicarius ornatus venoms, as well as Loxosceles venoms, were able to render erythrocytes susceptible to lysis by

  19. A new twist to the emerging functions of ceramides in cancer: novel role for platelet acid sphingomyelinase in cancer metastasis.

    PubMed

    Hannun, Yusuf A; Newcomb, Benjamin

    2015-06-01

    It is now appreciated that sphingolipids constitute a rich class of bioactive molecules that include ceramide, sphingosine, and sphingosine 1‐phosphate whose formation is controlled by a network of highly regulated enzymes (Hannun & Obeid, 2008). Notably, several stress stimuli induce the production of ceramide, which, as a single entity, has been traditionally associated with apoptotic and growth suppressive functions. However, recent data clearly suggest that this simplistic formulation is no longer tenable. PMID:25859016

  20. Neutral sphingomyelinase activity dependent on Mg2+ and anionic phospholipids in the intraerythrocytic malaria parasite Plasmodium falciparum.

    PubMed Central

    Hanada, K; Mitamura, T; Fukasawa, M; Magistrado, P A; Horii, T; Nishijima, M

    2000-01-01

    Sphingolipid metabolism and metabolites are important in various cellular events in eukaryotes. However, little is known about their function in plasmodial parasites. Here we demonstrate that neutral sphingomyelinase (SMase) involved in the sphingomyelin (SM) catabolism is retained by the intraerythrocytic parasite Plasmodium falciparum. When assayed in a neutral pH buffer supplemented with Mg(2+) and phosphatidylserine, an activity for the release of the phosphocholine group from SM was detected in parasite-infected, but not in uninfected, erythrocyte ghosts. The SMase activity in the parasite-infected erythrocyte ghosts was enhanced markedly by anionic phospholipids including unsaturated but not saturated phosphatidylserine. Mn(2+) could not substitute for Mg(2+) to activate SMase in parasite-infected erythrocyte ghosts, whereas both Mn(2+) and Mg(2+) activated mammalian neutral SMase. The specific activity level of SMase was higher in isolated parasites than in infected erythrocyte ghosts; further fractionation of lysates of the isolated parasites showed that the activity was bound largely to the membrane fraction of the parasites. The plasmodial SMase seemed not to hydrolyse phosphatidylcholine or phosphatidylinositol. The plasmodial SMase, but not SM synthase, was sensitive to scyphostatin, an inhibitor of mammalian neutral SMase, indicating that the plasmodial activities for SM hydrolysis and SM synthesis are mediated by different catalysts. Our finding that the malaria parasites possess SMase activity might explain why the parasites seem to have an SM synthase activity but no activity to synthesize ceramide de novo. PMID:10698693

  1. Increase in secretory sphingomyelinase activity and specific ceramides in the aorta of apolipoprotein E knockout mice during aging.

    PubMed

    Kobayashi, Keiko; Nagata, Eri; Sasaki, Kazuki; Harada-Shiba, Mariko; Kojo, Shosuke; Kikuzaki, Hiroe

    2013-01-01

    Atherosclerosis is caused by many factors, one of which is oxidative stress. We recently demonstrated that systemic oxidative stress increased secretory sphingomyelinase (sSMase) activity and generated ceramides in the plasma of diabetic rats. In addition, we also showed that the total ceramide level in human plasma correlated with the level of oxidized low-density lipoprotein. To investigate the relationship between ceramide species and atherogenesis during aging, we compared age-related changes in ceramide metabolism in apolipoprotein E knock out mice (apoE(-/-)) and wild type mice (WT). Although the total plasma ceramide level was higher in apoE(-/-) than that in WT at all ages, it decreased with increasing age. sSMase activity increased at 65 weeks (w) of age in both strains of mice. When apoE(-/-) developed atherosclerosis at 15 w of age, C18:0, C22:0, and C24:0 ceramide levels in the apoE(-/-) aorta significantly increased. Furthermore, at 65 w of age C16:0 and C24:1 ceramide levels were significantly higher than those in WT. These results suggested that elevation in levels of specific ceramide species due to sSMase activity contributed to atherogenesis during aging. PMID:23811568

  2. High-level over-expression, purification, and crystallization of a novel phospholipase C/sphingomyelinase from Pseudomonas aeruginosa

    PubMed Central

    Truan, Daphné; Vasil, Adriana; Stonehouse, Martin; Vasil, Michael L.; Pohl, Ehmke

    2013-01-01

    The hemolytic phospholipase C/sphingomyelinase PlcH from the opportunistic pathogen Pseudomonas aeruginosa represents the founding member of a growing family of virulence factors identified in a wide range of bacterial and fungal pathogens. In P. aeruginosa PlcH is co-expressed with a 17 kDa chaperone (PlcR2) and secreted as a fully folded heterodimer (PlcHR2) of approximately 95 kDa, by the twin arginine translocase (TAT) via the cytoplasmic membrane and through the outer membrane, by the Xcp (TypeII) secretory system. PlcHR2 has been shown to be an important virulence factor in model P. aeruginosa infections and is selectively cytotoxic, at picomolar concentrations to mammalian endothelial cells. Here we report how the various challenges starting from protein overexpression in the native organism P. aeruginosa, the use of detergents in the crystallization and data collection using the most advanced μ-focus synchrotron beam lines were overcome. Native diffraction data of this heterodimeric protein complex were collected up to a resolution of 4 Å, whereas needle-shaped crystals of l-selenomethionine substituted PlcHR2 with a maximum diameter of 10 micron were used to collect data sets with a maximum resolution of 2.75 Å. PMID:23201280

  3. A methodology for existing system upgrade to current ASME standards and system lifetime extension

    SciTech Connect

    Burr, T.K.; Dwight, J.E. Jr.; Hawkes, G.L.; Pace, N.E.

    1990-01-01

    In the wake of the Chernobyl events, there has been an increase in the awareness and review of government operated reactors both internationally, and within the United States. Government reactors have recently come under increased and indepth scrutiny. Department of Energy Secretary Hodel committed to a review of the safety of non-commercial reactors and irradiation facilities within the department. The increased attention has been in the areas of accident response, PRA of the facilities, environmental impacts, and the construction and associated standards for the facilities. This paper focuses on the system qualifications to current standards. Specifically, this paper discusses a method used for upgrading an existing high pressure nuclear system to current ASME Code standards and to extend the system's lifetime. This paper reports the methods used in an attempt to qualify components of the Advanced Test Reactor (ATR) located at the Idaho National Engineering Laboratory (INEL) to current ASME Code Section III standards.

  4. ASME AG-1 Section FC Qualified HEPA Filters; a Particle Loading Comparison - 13435

    SciTech Connect

    Stillo, Andrew; Ricketts, Craig I.

    2013-07-01

    High Efficiency Particulate Air (HEPA) Filters used to protect personnel, the public and the environment from airborne radioactive materials are designed, manufactured and qualified in accordance with ASME AG-1 Code section FC (HEPA Filters) [1]. The qualification process requires that filters manufactured in accordance with this ASME AG-1 code section must meet several performance requirements. These requirements include performance specifications for resistance to airflow, aerosol penetration, resistance to rough handling, resistance to pressure (includes high humidity and water droplet exposure), resistance to heated air, spot flame resistance and a visual/dimensional inspection. None of these requirements evaluate the particle loading capacity of a HEPA filter design. Concerns, over the particle loading capacity, of the different designs included within the ASME AG-1 section FC code[1], have been voiced in the recent past. Additionally, the ability of a filter to maintain its integrity, if subjected to severe operating conditions such as elevated relative humidity, fog conditions or elevated temperature, after loading in use over long service intervals is also a major concern. Although currently qualified HEPA filter media are likely to have similar loading characteristics when evaluated independently, filter pleat geometry can have a significant impact on the in-situ particle loading capacity of filter packs. Aerosol particle characteristics, such as size and composition, may also have a significant impact on filter loading capacity. Test results comparing filter loading capacities for three different aerosol particles and three different filter pack configurations are reviewed. The information presented represents an empirical performance comparison among the filter designs tested. The results may serve as a basis for further discussion toward the possible development of a particle loading test to be included in the qualification requirements of ASME AG-1

  5. A method for certification of FRP piping fabricators for ASME B31.3 systems

    SciTech Connect

    Andersen, K.D.

    1996-07-01

    Cost-effective FRP piping is often the material of choice for transport of corrosive chemicals. Plant Managers and Engineers have great concern about the integrity of FRP piping joints and the safety of these systems. A specification requirement, in the bid documents, that all fabricators be Certified by the FRP piping manufacturer is a method to promote successful fabrication. A method is proposed, which is in accordance with ASME B31.3 Piping Code, to train and certify fabricators.

  6. A Proposal for the Maximum KIC for Use in ASME Code Flaw and Fracture Toughness Evaluations

    SciTech Connect

    Kirk, Mark; Stevens, Gary; Erickson, Marjorie A; Yin, Shengjun

    2011-01-01

    Nonmandatory Appendices A [1] and G [2] of Section XI of the ASME Code use the KIc curve (indexed to the material reference transition temperature, RTNDT) in reactor pressure vessel (RPV) flaw evaluations, and for the purpose of establishing RPV pressure-temperature (P-T) limits. Neither of these appendices places an upper-limit on the KIc value that may be used in these assessments. Over the years, it has often been suggested by some of the members of the ASME Section XI Code committees that are responsible for maintaining Appendices A and G that there is a practical upper limit of 200 ksi in (220 MPa m) [4]. This upper limit is not well recognized by all users of the ASME Code, is not explicitly documented within the Code itself, and the one source known to the authors where it is defended [4] relies on data that is either in error, or is less than 220 MPa m. However, as part of the NRC/industry pressurized thermal shock (PTS) re-evaluation effort, empirical models were developed that propose common temperature dependencies for all ferritic steels operating on the upper shelf. These models relate the fracture toughness properties in the transition regime to those on the upper shelf and, combined with data for a wide variety of RPV steels and welds on which they are based, suggest that the practical upper limit of 220 MPa m exceeds the upper shelf fracture toughness of most RPV steels by a considerable amount, especially for irradiated steels. In this paper, available models and data are used to propose upper bound limits of applicability on the KIc curve for use in ASME Code, Section XI, Nonmandatory Appendices A and G evaluations that are consistent with available data for RPV steels.

  7. Need for ASME code changes for reliable characterization of thermal fatigue cracks in Class 1 components

    SciTech Connect

    Shah, V.N.; Ware, A.G.

    1991-01-01

    This paper evaluates the current inspection requirements for detecting and sizing thermal fatigue cracks in major light water reactor components. A review of field experience indicates that several sites in base metal that are susceptible to thermal fatigue damage are not included in the service inspection programs. Current inservice inspection procedures and conventional ultrasonic inspection techniques are not adequate for reliable detection and sizing of thermal fatigue cracks. This paper makes several recommendations for changes in ASME Section 11 inspection requirements. 12 refs.

  8. RXTE/ASM Observations Of SS 433 And Cygnus X-2

    NASA Astrophysics Data System (ADS)

    Hoffman, Lisa; Mason, P. A.

    2010-01-01

    We present a dynamic period search analysis of the X-ray binaries SS 433 and Cygnus X-2 using data from the Rossi X-Ray Timing Explorer All Sky Monitor (RXTE/ASM) spanning over 13 years. We report the detection of a period in SS 433 near 162 days. This may be the first detection of the disk precession period in X-rays. We detect an 81.8 day period in the object Cygnus X-2. The RXTE/ASM light curve is inconsistent with the 77.7 day X-ray period of Wijnands et al. (1996), which was based on a small subset of the RXTE/ASM data combined with data from VELA 5B, and Ariel 5 All-Sky Monitors. Since Cygnus X-2 displays periodic behavior that seems to come and go, producing different best-fit periods on time scales of a few years; we suggest that Cygnus X-2 exhibits quasi-periodic oscillations of about 80 days. This research is supported by a grant from the New Mexico Space Grant Consortium.

  9. A 2015 Igrf Candidate Model Based on Swarm's Experimental ASM Vector Mode Data

    NASA Astrophysics Data System (ADS)

    Vigneron, P.; Hulot, G.; Olsen, N.; Leger, J. M.; Jager, T.; Brocco, L.; Sirol, O.; Coisson, P.; Lalanne, X.; Chulliat, A.; Bertrand, F.; Boness, A.; Fratter, I.

    2014-12-01

    Each of the three Alpha, Bravo and Charlie satellites of the ESA Swarm mission carries an Absolute Scalar Magnetometer (CNES customer furnished ASM instrument designed by CEA-Léti) that provides the nominal 1 Hz scalar data of the mission, but also delivers 1 Hz experimental vector data. Tests during the commissioning and calibration/validation phase have shown that these data and the rigidity of the boom mechanically linking the ASM to the star imager (STR) on Alpha and Bravo were of such good quality that an IGRF candidate geomagnetic field model could possibly be produced from such ASM-only data (without having to resort to any of the nominal vector field magnetometer (VFM) data of the mission). In this presentation, we will report on our efforts to build such an IGRF candidate, which intends to provide an image of the January 1, 2015 Geomagnetic Field, alternative to the images provided by IGRF candidate models based on Swarm nominal L1b data, or other data.

  10. Staphylococcus aureus β-Toxin Mutants Are Defective in Biofilm Ligase and Sphingomyelinase Activity, and Causation of Infective Endocarditis and Sepsis.

    PubMed

    Herrera, Alfa; Vu, Bao G; Stach, Christopher S; Merriman, Joseph A; Horswill, Alexander R; Salgado-Pabón, Wilmara; Schlievert, Patrick M

    2016-05-01

    β-Toxin is an important virulence factor of Staphylococcus aureus, contributing to colonization and development of disease [Salgado-Pabon, W., et al. (2014) J. Infect. Dis. 210, 784-792; Huseby, M. J., et al. (2010) Proc. Natl. Acad. Sci. U.S.A. 107, 14407-14412; Katayama, Y., et al. (2013) J. Bacteriol. 195, 1194-1203]. This cytotoxin has two distinct mechanisms of action: sphingomyelinase activity and DNA biofilm ligase activity. However, the distinct mechanism that is most important for its role in infective endocarditis is unknown. We characterized the active site of β-toxin DNA biofilm ligase activity by examining deficiencies in site-directed mutants through in vitro DNA precipitation and biofilm formation assays. Possible conformational changes in mutant structure compared to that of wild-type toxin were assessed preliminarily by trypsin digestion analysis, retention of sphingomyelinase activity, and predicted structures based on the native toxin structure. We addressed the contribution of each mechanism of action to producing infective endocarditis and sepsis in vivo in a rabbit model. The H289N β-toxin mutant, lacking sphingomyelinase activity, exhibited lower sepsis lethality and infective endocarditis vegetation formation compared to those of the wild-type toxin. β-Toxin mutants with disrupted biofilm ligase activity did not exhibit decreased sepsis lethality but were deficient in infective endocarditis vegetation formation compared to the wild-type protein. Our study begins to characterize the DNA biofilm ligase active site of β-toxin and suggests β-toxin functions importantly in infective endocarditis through both of its mechanisms of action. PMID:27015018

  11. Understanding the Long-Term Spectral Variability of Cygnus X-1 from BATSE and ASM Observations

    NASA Technical Reports Server (NTRS)

    Zdziarski, Andrzej A.; Poutanen, Juri; Paciesas, William S.; Wen, Linqing; Six, N. Frank (Technical Monitor)

    2002-01-01

    We present a spectral analysis of observations of Cygnus X-1 by the RXTE/ASM (1.5-12 keV) and CGRO/BATSE (20-300 keV), including about 1200 days of simultaneous data. We find a number of correlations between intensities and hardnesses in different energy bands from 1.5 keV to 300 keV. In the hard (low) spectral state, there is a negative correlation between the ASM 1.5-12 keV flux and the hardness at any energy. In the soft (high) spectral state, the ASM flux is positively correlated with the ASM hardness (as previously reported) but uncorrelated with the BATSE hardness. In both spectral states, the BATSE hardness correlates with the flux above 100 keV, while it shows no correlation with the flux in the 20-100 keV range. At the same time, there is clear correlation between the BATSE fluxes below and above 100 keV. In the hard state, most of the variability can be explained by softening the overall spectrum with a pivot at approximately 50 keV. The observations show that there has to be another, independent variability pattern of lower amplitude where the spectral shape does not change when the luminosity changes. In the soft state, the variability is mostly caused by a variable hard (Comptonized) spectral component of a constant shape superimposed on a constant soft blackbody component. These variability patterns are in agreement with the dependence of the rms variability on the photon energy in the two states. We interpret the observed correlations in terms of theoretical Comptonization models. In the hard state, the variability appears to be driven mostly by changing flux in seed photons Comptonized in a hot thermal plasma cloud with an approximately constant power supply. In the soft state, the variability is consistent with flares of hybrid, thermal/nonthermal, plasma with variable power above a stable cold disk. Also, based on broadband pointed observations simultaneous with those of the ASM and BATSE, we find the intrinsic bolometric luminosity increases by a

  12. The essential neutral sphingomyelinase is involved in the trafficking of the variant surface glycoprotein in the bloodstream form of Trypanosoma brucei

    PubMed Central

    Young, Simon A; Smith, Terry K

    2010-01-01

    Sphingomyelin is the main sphingolipid in Trypanosoma brucei, the causative agent of African sleeping sickness. In vitro and in vivo characterization of the T. brucei neutral sphingomyelinase demonstrates that it is directly involved in sphingomyelin catabolism. Gene knockout studies in the bloodstream form of the parasite indicate that the neutral sphingomyelinase is essential for growth and survival, thus highlighting that the de novo biosynthesis of ceramide is unable to compensate for the loss of sphingomyelin catabolism. The phenotype of the conditional knockout has given new insights into the highly active endocytic and exocytic pathways in the bloodstream form of T. brucei. Hence, the formation of ceramide in the endoplasmic reticulum affects post-Golgi sorting and rate of deposition of newly synthesized GPI-anchored variant surface glycoprotein on the cell surface. This directly influences the corresponding rate of endocytosis, via the recycling endosomes, of pre-existing cell surface variant surface glycoprotein. The trypanosomes use this coupled endocytic and exocytic mechanism to maintain the cell density of its crucial variant surface glycoprotein protective coat. TbnSMase is therefore genetically validated as a drug target against African trypanosomes, and suggests that interfering with the endocytic transport of variant surface glycoprotein is a highly desirable strategy for drug development against African trypanosomasis. PMID:20398210

  13. The ASM-NSF Biology Scholars Program: An Evidence-Based Model for Faculty Development.

    PubMed

    Chang, Amy L; Pribbenow, Christine M

    2016-05-01

    The American Society for Microbiology (ASM) established its ASM-NSF (National Science Foundation) Biology Scholars Program (BSP) to promote undergraduate education reform by 1) supporting biologists to implement evidence-based teaching practices, 2) engaging life science professional societies to facilitate biologists' leadership in scholarly teaching within the discipline, and 3) participating in a teaching community that fosters disciplinary-level science, technology, engineering, and mathematics (STEM) reform. Since 2005, the program has utilized year-long residency training to provide a continuum of learning and practice centered on principles from the scholarship of teaching and learning (SoTL) to more than 270 participants ("scholars") from biology and multiple other disciplines. Additionally, the program has recruited 11 life science professional societies to support faculty development in SoTL and discipline-based education research (DBER). To identify the BSP's long-term outcomes and impacts, ASM engaged an external evaluator to conduct a study of the program's 2010-2014 scholars (n = 127) and society partners. The study methods included online surveys, focus groups, participant observation, and analysis of various documents. Study participants indicate that the program achieved its proposed goals relative to scholarship, professional society impact, leadership, community, and faculty professional development. Although participants also identified barriers that hindered elements of their BSP participation, findings suggest that the program was essential to their development as faculty and provides evidence of the BSP as a model for other societies seeking to advance undergraduate science education reform. The BSP is the longest-standing faculty development program sponsored by a collective group of life science societies. This collaboration promotes success across a fragmented system of more than 80 societies representing the life sciences and helps

  14. The ASM-NSF Biology Scholars Program: An Evidence-Based Model for Faculty Development

    PubMed Central

    Chang, Amy L.; Pribbenow, Christine M.

    2016-01-01

    The American Society for Microbiology (ASM) established its ASM-NSF (National Science Foundation) Biology Scholars Program (BSP) to promote undergraduate education reform by 1) supporting biologists to implement evidence-based teaching practices, 2) engaging life science professional societies to facilitate biologists’ leadership in scholarly teaching within the discipline, and 3) participating in a teaching community that fosters disciplinary-level science, technology, engineering, and mathematics (STEM) reform. Since 2005, the program has utilized year-long residency training to provide a continuum of learning and practice centered on principles from the scholarship of teaching and learning (SoTL) to more than 270 participants (“scholars”) from biology and multiple other disciplines. Additionally, the program has recruited 11 life science professional societies to support faculty development in SoTL and discipline-based education research (DBER). To identify the BSP’s long-term outcomes and impacts, ASM engaged an external evaluator to conduct a study of the program’s 2010–2014 scholars (n = 127) and society partners. The study methods included online surveys, focus groups, participant observation, and analysis of various documents. Study participants indicate that the program achieved its proposed goals relative to scholarship, professional society impact, leadership, community, and faculty professional development. Although participants also identified barriers that hindered elements of their BSP participation, findings suggest that the program was essential to their development as faculty and provides evidence of the BSP as a model for other societies seeking to advance undergraduate science education reform. The BSP is the longest-standing faculty development program sponsored by a collective group of life science societies. This collaboration promotes success across a fragmented system of more than 80 societies representing the life sciences and

  15. New methods of analysis of materials strength data for the ASME Boiler and Pressure Vessel Code

    SciTech Connect

    Booker, M.K.; Booker, B.L.P.

    1980-01-01

    Tensile and creep data of the type used to establish allowable stress levels for the ASME Boiler and Pressure Vessel Code have been examined for type 321H stainless steel. Both inhomogeneous, unbalanced data sets and well-planned homogeneous data sets have been examined. Data have been analyzed by implementing standard manual techniques on a modern digital computer. In addition, more sophisticated techniques, practical only through the use of the computer, have been applied. The result clearly demonstrates the efficacy of computerized techniques for these types of analyses.

  16. ASME N510 test results for Savannah River Site AACS filter compartments

    SciTech Connect

    Paul, J.D.; Punch, T.M.

    1995-02-01

    The K-Reactor at the Savannah River Site recently implemented design improvements for the Airborne Activity Confinement System (AACS) by procuring, installing, and testing new Air Cleaning Units, or filter compartments, to ASME AG-11, N509, and N510 requirements. Specifically, these new units provide documentable seismic resistance to a Design Basis Accident earthquake, provide 2 inch adsorber beds with 0.25 second residence time, and meet all AG-1, N509, and N510 requirements for testability and maintainability. This paper presents the results of the Site acceptance testing and discusses an issue associated with sample manifold qualification testing.

  17. Material organizations in the 1994 Addenda ASME Code Section III, Division I

    SciTech Connect

    Bressler, M.N.; Kist, N.C.

    1995-12-01

    This paper provides a history of the requirements for materials in ASME Code Section 3, Nuclear Power Plant Components, from its inception to the present 1992 Edition and the new 1994 Addenda. Major events in the development of the Code are listed, and the organizations involved in materials that are covered by this standard arc described. Changes to duties and responsibilities of individuals and organizations performing Section 3 material activities are identified, and the requirements for accreditation and qualification are discussed. A Code Case to provide alternatives to compliance is presented.

  18. Friction and wear testing source book: Selected references from ASTM standards and ASM handbooks

    SciTech Connect

    1997-12-31

    A collection of key standards and reference articles about friction, lubrication, and wear makes this a valuable reference book to answer questions about wear mechanisms and testing, and how different measurement methods compare to each other. The focus is on test methods, with supporting material on analysis of wear-related failures. Selected information is reprinted from ASTM Standards, Volumes 2.05, 3.02, 5.02, 5.03 and 13.01, and ASM Handbooks, Volumes 8 (Mechanical Testing), 11 (Failure Analysis and Prevention) and 18 (Friction, Lubrication, and Wear Technology).

  19. Materials and design bases issues in ASME Code Case N-47

    SciTech Connect

    Huddleston, R.L.; Swindeman, R.W. )

    1993-04-01

    A preliminary evaluation of the design bases (principally ASME Code Case N-47) was conducted for design and operation of reactors at elevated temperatures where the time-dependent effects of creep, creep-fatigue, and creep ratcheting are significant. Areas where Code rules or regulatory guides may be lacking or inadequate to ensure the operation over the expected life cycles for the next-generation advanced high-temperature reactor systems, with designs to be certified by the US Nuclear Regulatory Commission, have been identified as unresolved issues. Twenty-two unresolved issues were identified and brief scoping plans developed for resolving these issues.

  20. Overview of the new ASME Performance Test Code for wind turbines

    NASA Technical Reports Server (NTRS)

    Spera, David A.

    1986-01-01

    The principal technical features of the ASME Performance Test Code for wind turbines are presented and such issues as what sizes and types of wind turbines should be included, what the principal measure of performance should be, and how wind speed should be measured are discussed. It is concluded that the present test code is applicable to wind turbine systems of all sizes. The principal measure of performance as defined by this code is net energy output and the primary performance parameter is the 'test energy ratio' which is based on a comparison between the measured and predicted energy output for the test period.

  1. Application of the ASME code in the design of the GA-4 and GA-9 casks

    SciTech Connect

    Mings, W.J. ); Koploy, M.A. )

    1992-01-01

    General Atomics (GA) is developing two spent fuel shipping casks for transport by legal weight truck (LWT). The casks are designed to the loading, environmental conditions and safety requirements defined in Title 10 of the Code of Federal Regulations, Part 71 (10CFR71). To ensure that all components of the cask meet the 10CFR71 rules, GA established structural design criteria for each component based on NRC Regulatory Guides and the American Society of Mechanical Engineers Boiler and Pressure Vessel Code (ASME Code). This paper discusses the criteria used for different cask components, how they were applied and the conservatism and safety margins built into the criteria and assumption.

  2. Application of the ASME code in the design of the GA-4 and GA-9 casks

    SciTech Connect

    Mings, W.J.; Koploy, M.A.

    1992-08-01

    General Atomics (GA) is developing two spent fuel shipping casks for transport by legal weight truck (LWT). The casks are designed to the loading, environmental conditions and safety requirements defined in Title 10 of the Code of Federal Regulations, Part 71 (10CFR71). To ensure that all components of the cask meet the 10CFR71 rules, GA established structural design criteria for each component based on NRC Regulatory Guides and the American Society of Mechanical Engineers Boiler and Pressure Vessel Code (ASME Code). This paper discusses the criteria used for different cask components, how they were applied and the conservatism and safety margins built into the criteria and assumption.

  3. Application of the ASME code in designing containment vessels for packages used to transport radioactive materials

    SciTech Connect

    Raske, D.T.; Wang, Z.

    1992-07-01

    The primary concern governing the design of shipping packages containing radioactive materials is public safety during transport. When these shipments are within the regulatory jurisdiction of the US Department of Energy, the recommended design criterion for the primary containment vessel is either Section III or Section VIII, Division 1, of the ASME Boiler and Pressure Vessel Code, depending on the activity of the contents. The objective of this paper is to discuss the design of a prototypic containment vessel representative of a packaging for the transport of high-level radioactive material.

  4. Quasi-ellipsoidal heads to ASME Code under internal pressure stress analysis by the Force Method

    SciTech Connect

    Guedes, E.

    1996-12-01

    Torispherically-dished pressure vessel heads that meet Section VIII of the ASME Code can be completely analyzed by means of the so-called Force Method. Thus combined normal stresses (pressure plus radial growth plus bending) at the shell junctures that make up those heads may be determined to a good degree of accuracy, along with the ideal bending-free location of the welding line (circumferential joint that connects head to shell). As a useful consequence, head skirt is kept to a safe minimum.

  5. Code cases for implementing risk-based inservice testing in the ASME OM code

    SciTech Connect

    Rowley, C.W.

    1996-12-01

    Historically inservice testing has been reasonably effective, but quite costly. Recent applications of plant PRAs to the scope of the IST program have demonstrated that of the 30 pumps and 500 valves in the typical plant IST program, less than half of the pumps and ten percent of the valves are risk significant. The way the ASME plans to tackle this overly-conservative scope for IST components is to use the PRA and plant expert panels to create a two tier IST component categorization scheme. The PRA provides the quantitative risk information and the plant expert panel blends the quantitative and deterministic information to place the IST component into one of two categories: More Safety Significant Component (MSSC) or Less Safety Significant Component (LSSC). With all the pumps and valves in the IST program placed in MSSC or LSSC categories, two different testing strategies will be applied. The testing strategies will be unique for the type of component, such as centrifugal pump, positive displacement pump, MOV, AOV, SOV, SRV, PORV, HOV, CV, and MV. A series of OM Code Cases are being developed to capture this process for a plant to use. One Code Case will be for Component Importance Ranking. The remaining Code Cases will develop the MSSC and LSSC testing strategy for type of component. These Code Cases are planned for publication in early 1997. Later, after some industry application of the Code Cases, the alternative Code Case requirements will gravitate to the ASME OM Code as appendices.

  6. Results from Evaluation of Proposed ASME AG-1 Section FI Metal Media Filters - 13063

    SciTech Connect

    Wilson, John A.; Giffin, Paxton K.; Parsons, Michael S.; Waggoner, Charles A.

    2013-07-01

    High efficiency particulate air (HEPA) filtration technology is commonly used in Department of Energy (DOE) facilities that require control of radioactive particulate matter (PM) emissions due to treatment or management of radioactive materials. Although HEPA technology typically makes use of glass fiber media, metal and ceramic media filters are also capable of filtering efficiencies beyond the required 99.97%. Sintered metal fiber filters are good candidates for use in DOE facilities due to their resistance to corrosive environments and resilience at high temperature and elevated levels of relative humidity. Their strength can protect them from high differential pressure or pressure spikes and allow for back pulse cleaning, extending filter lifetime. Use of these filters has the potential to reduce the cost of filtration in DOE facilities due to life cycle cost savings. ASME AG-1 section FI has not been approved due to a lack of protocols and performance criteria for qualifying section FI filters. The Institute for Clean Energy Technology (ICET) with the aid of the FI project team has developed a Section FI test stand and test plan capable of assisting in the qualification ASME AG-1 section FI filters. Testing done at ICET using the FI test stand evaluates resistance to rated air flow, test aerosol penetration and resistance to heated air of the section FI filters. Data collected during this testing consists of temperature, relative humidity, differential pressure, flow rate, upstream particle concentration, and downstream particle concentration. (authors)

  7. A strategy for implementation of experience based seismic equipment qualification in IEEE and ASME industry standards

    SciTech Connect

    Adams, T.M.

    1996-12-01

    In the past 20 years, extensive data on the performance of mechanical and electric equipment during actual strong motion earthquakes and seismic qualification tests has been accumulated. Recognizing that an experience based approach provides a technically sound and cost effective method for the seismic qualification of some or certain equipment, the IEEE Nuclear Power Engineering Committee and the ASME Committee on Qualification of Mechanical Equipment established a Special Working Group to investigate the incorporation of experienced based methods into the industry consensus codes and standards currently used in the seismic qualification of Seismic Category Nuclear Power Plant equipment. This paper presents the strategy (course of action) which was developed by the Special Working Group for meeting this objective of incorporation of experience based seismic qualification standards used in the design and seismic qualification of seismic category nuclear power plant equipment. This strategy was recommended to both chartering organizations, the IEEE Nuclear Power Engineering Committee and the ASME Committee on Qualification of Mechanical Equipment for their consideration and implementation. The status of the review and implementation of the Special Working Group`s recommended strategy by the sponsoring organization is also discussed.

  8. Methods for incorporating effects of LWR coolant environment into ASME code fatigue evaluations.

    SciTech Connect

    Chopra, O. K.

    1999-04-15

    The ASME Boiler and Pressure Vessel Code provides rules for the construction of nuclear power plant components. Appendix I to Section HI of the Code specifies design fatigue curves for structural materials. However, the effects of light water reactor (LWR) coolant environments are not explicitly addressed by the Code design curves. Recent test data illustrate potentially significant effects of LWR environments on the fatigue resistance of carbon and low-alloy steels and austenitic stainless steels (SSs). Under certain loading and environmental conditions, fatigue lives of carbon and low-alloy steels can be a factor of {approx}70 lower in an LWR environment than in air. These results raise the issue of whether the design fatigue curves in Section III are appropriate for the intended purpose. This paper presents the two methods that have been proposed for incorporating the effects of LWR coolant environments into the ASME Code fatigue evaluations. The mechanisms of fatigue crack initiation in carbon and low-alloy steels and austenitic SSs in LWR environments are discussed.

  9. Overview of new rules and recent changes in ASME code, Section VIII, pressure vessels

    SciTech Connect

    Farr, J.R.

    1995-12-01

    In this presentation, some of the new rules and recent changes to the ASME Boiler and Pressure Vessel Code, Section 8, Divisions 1 and 2, are reviewed. On July 1, 1995, the 1995 Edition of the ASME Code was issued. This 1995 Edition incorporates those items which were added of changed in the 1992, 1993, and 1994 Addenda to the 1992 Edition of the Code. The 1995 Edition contains no new items which were not included in the previous edition and three addenda. With the possibility of an extended time before some of the new rules are able to appear in the addenda, the recent trend is to put the rules in Code Cases which are approved earlier. Consequently, it is necessary to review new Code Cases as well as Code changes. Updates continue for impact requirements for standard components as well as for materials other than UCS, carbon steel and low alloys. Extensive changes have been made for UHA, high-alloy, materials regarding impact requirements. Example problems have been revised to include these effects. Significant changes are reviewed.

  10. Comparison of ASME Code NB-3200 and NB-3600 results for fatigue analysis of B31.1 branch nozzles

    SciTech Connect

    Nitzel, M.E.; Ware, A.G.; Morton, D.K.

    1996-06-01

    Fatigue analyses wre conducted on two reactor coolant system branch nozzles in an operating PWR designed to the B31.1 Code, for which no explicit fatigue analysis was required by the licensing basis. These analyses were performed as part of resolving issues connected with NRC`s Fatigue Action Plan to determine if the cumulative usage factor (CUF) for these nozzles, using the 1992 ASME Code and representative PWR transients, were comparable to nozzles designed and analyzed to the ASME Code. Both NB-3200 and NB-3600 ASME Code methods were used. NB-3200 analyses included the development of finite element models for each nozzle. Although detailed thermal transients were not available for the plant analyzed, representative transients from similar PWRs were applied in each method. CUFs calculated using NB-3200 methods were significantly less than using NB-3600. The paper points out differences in analysis methods and highlights difficulties and unknowns in performing more detailed analyses to reduce conservative assumptions.

  11. 3D automatic anatomy segmentation based on iterative graph-cut-ASM

    SciTech Connect

    Chen, Xinjian; Bagci, Ulas

    2011-08-15

    Purpose: This paper studies the feasibility of developing an automatic anatomy segmentation (AAS) system in clinical radiology and demonstrates its operation on clinical 3D images. Methods: The AAS system, the authors are developing consists of two main parts: object recognition and object delineation. As for recognition, a hierarchical 3D scale-based multiobject method is used for the multiobject recognition task, which incorporates intensity weighted ball-scale (b-scale) information into the active shape model (ASM). For object delineation, an iterative graph-cut-ASM (IGCASM) algorithm is proposed, which effectively combines the rich statistical shape information embodied in ASM with the globally optimal delineation capability of the GC method. The presented IGCASM algorithm is a 3D generalization of the 2D GC-ASM method that they proposed previously in Chen et al.[Proc. SPIE, 7259, 72590C1-72590C-8 (2009)]. The proposed methods are tested on two datasets comprised of images obtained from 20 patients (10 male and 10 female) of clinical abdominal CT scans, and 11 foot magnetic resonance imaging (MRI) scans. The test is for four organs (liver, left and right kidneys, and spleen) segmentation, five foot bones (calcaneus, tibia, cuboid, talus, and navicular). The recognition and delineation accuracies were evaluated separately. The recognition accuracy was evaluated in terms of translation, rotation, and scale (size) error. The delineation accuracy was evaluated in terms of true and false positive volume fractions (TPVF, FPVF). The efficiency of the delineation method was also evaluated on an Intel Pentium IV PC with a 3.4 GHZ CPU machine. Results: The recognition accuracies in terms of translation, rotation, and scale error over all organs are about 8 mm, 10 deg. and 0.03, and over all foot bones are about 3.5709 mm, 0.35 deg. and 0.025, respectively. The accuracy of delineation over all organs for all subjects as expressed in TPVF and FPVF is 93.01% and 0.22%, and

  12. Strong Inhibition of Secretory Sphingomyelinase by Catechins, Particularly by (-)-Epicatechin 3-O-Gallate and (-)-3'-O-Methylepigallocatechin 3-O-Gallate.

    PubMed

    Kobayashi, Keiko; Ishizaki, Yuki; Kojo, Shosuke; Kikuzaki, Hiroe

    2016-01-01

    Sphingomyelinases (SMases) are key enzymes involved in many diseases which are caused by oxidative stress, such as atherosclerosis, diabetes mellitus, nonalcoholic fatty liver disease, and Alzheimer's disease. SMases hydrolyze sphingomyelin to generate ceramide, a well-known pro-apoptotic lipid. SMases are classified into five types based on pH optimum, subcellular localization, and cation dependence. Previously, we demonstrated that elevation of secretory sphingomyelinase (sSMase) activity increased the plasma ceramide concentration under oxidative stress induced by diabetes and atherosclerosis in murine models. These results suggest that sSMase inhibitors can prevent the progress of these diseases. The present study demonstrated that sSMase activity was activated by oxidation and inhibited by reduction. Furthermore, we examined whether catechins inhibited the sSMase activity in a physiological plasma concentration. Among catechins, (-)-epicatechin 3-O-gallate (ECg) exhibited strong inhibitory effect on sSMase (IC50=25.7 μM). This effect was attenuated by methylation at the 3″- or 4″-position. On the other hand, (-)-epigallocatechin 3-O-gallate (EGCg) and (-)-catechin 3-O-gallate (Cg) exhibited weaker inhibitory activity than ECg, and (-)-epicatechin and (-)-epigallocatechin did not affect sSMase activity. Additionally, one synthetic catechin, (-)-3'-O-methylepigallocatechin 3-O-gallate (EGCg-3'-O-Me), showed the strongest inhibitory effect (IC50=1.7 μM) on sSMase. This phenomenon was not observed for (-)-4'-O-methylepigallocatechin 3-O-gallate. These results suggest that the reduction potential, the presence of the galloyl residue at the C-3 position, and the steric requirement to interact with sSMase protein are important for effective inhibition of sSMase. PMID:27264097

  13. Crack growth in ASME SA-105 grade 2 steel in hydrogen at ambient temperature

    NASA Technical Reports Server (NTRS)

    Walter, R. J.

    1975-01-01

    Cyclic-load crack growth measurements were performed on ASME SA-105 Grade 2 steel specimens exposed to 10,000- and 15,000-psi hydrogen and to 5000-psi helium, all at ambient temperatures. The cyclic-load crack growth rate was found to be faster in high-pressure hydrogen than in helium. Cyclic-load crack growth rates in this steel were not reduced by preloading in air to a stress intensity of 1.5 times the cyclic K sub max in hydrogen. There are indications that holding under load in hydrogen, and loading and unloading in helium retards hydrogen-accelerated cyclic-load crack growth. Cyclic frequency and R (ratio of K sub min/k sub max) were important variables determining crack growth rate. The crack growth rate increased as a logarithm of the cycle duration and decreased with increasing R.

  14. Investigation of ASME code: Section 3, Subsection NB, Suggested revisions: Final report

    SciTech Connect

    Love, J.E.

    1986-11-01

    The nuclear industry has become increasingly aware that engineering standards documents include requirements which are technically inconsistent and in many cases excessively costly to apply. As a result of these concerns, a study of Subsection NB of Section III of the ASME Code was undertaken. Subsection NB addresses Code requirements for Class 1 Components for nuclear power plants. The study was restricted to one subsection with the intent of discovering the extent of the difficulties in the Code. The areas of primary concern were those requirements which have a direct impact on design, fabrication, and examination. Subsection NB was carefully reviewed for inconsistencies and unworkable criteria. Seventy-seven such deficiencies were identified. A preliminary recommendation was prepared for each inconsistency or unworkable criterion. An overall critique of the Subsection was written, and suggestions were made for addressing the problems as part of a major Code revision.

  15. ASM conference report: genetics and molecular biology of industrial microorganisms 1984

    SciTech Connect

    Not Available

    1985-01-01

    The ASM Conference on Genetics and Molecular Biology of Industrial Microorganisms was held Sept. 30 to Oct. 3, 1984 in Bloomington, Indiana. The meetings reflected the progress in applying molecular techniques to the genetic analysis of industrial microorganisms. Topics of discussion included; gene expression in yeasts; the cloning of an Aspergillus sp. gene for glucomylase into S. cerevisiae; the construction of a yeast acentric ring plasmid (YARpl); the cloning of hygromycin resistant genes into Cephalosporium; optimization of gene expression in E. coli; a model for the initiation of translation in E. coli based on experiments with T4rIIb mutants; the role of proteases in protein turnover; evidence indicating which segments on RNAs are needed for the initiation of DNA synthesis; the application of various gene expression systems for the production of vaccines; the sporulation genes of Bacillus; the inducible chloramphenicol resistance found in Bacillus; gene expression in Streptomyces; enzyme activities in Streptomyces; and cloning of genes involved in antibiotic biosynthesis.

  16. Calibration and simulation of ASM2d at different temperatures in a phosphorus removal pilot plant.

    PubMed

    García-Usach, F; Ferrer, J; Bouzas, A; Seco, A

    2006-01-01

    In this work, an organic and nutrient removal pilot plant was used to study the temperature influence on phosphorus accumulating organisms. Three experiments were carried out at 13, 20 and 24.5 degrees C, achieving a high phosphorus removal percentage in all cases. The ASM2d model was calibrated at 13 and 20 degrees C and the Arrhenius equation constant was obtained for phosphorus removal processes showing that the temperature influences on the biological phosphorus removal subprocesses in a different degree. The 24.5 degrees C experiment was simulated using the model parameters obtained by means of the Arrhenius equation. The simulation results for the three experiments showed good correspondence with the experimental data, demonstrating that the model and the calibrated parameters were able to predict the pilot plant behaviour. PMID:16889256

  17. Change of nonlinear acoustics in ASME grade 122 steel welded joint during creep

    NASA Astrophysics Data System (ADS)

    Ohtani, Toshihiro; Honma, Takumi; Ishii, Yutaka; Tabuchi, Masaaki; Hongo, Hiromichi; Hirao, Masahiko

    2016-02-01

    In this paper, we described the changes of two nonlinear acoustic characterizations; resonant frequency shift and three-wave interaction, with electromagnetic acoustic resonance (EMAR) throughout the creep life in the welded joints of ASME Grade 122, one of high Cr ferritic heat resisting steels. EMAR was a combination of the resonant acoustic technique with a non-contact electromagnetic acoustic transducer (EMAT). These nonlinear acoustic parameters decreased from the start to 50% of creep life. After slightly increased, they rapidly increased from 80% of creep life to rupture. We interpreted these phenomena in terms of dislocation recovery, recrystallization, and restructuring related to the initiation and growth of creep void, with support from the SEM and TEM observation.

  18. 76 FR 11191 - Hazardous Materials: Adoption of ASME Code Section XII and the National Board Inspection Code

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-03-01

    ... the ANPRM published on December 23, 2010 (ANPRM; 75 FR 80765). The ASME Boiler and Pressure Vessel... April 11, 2000 (65 FR 19477) or you may visit http://www.regulations.gov . SUPPLEMENTARY INFORMATION: I. Background On December 23, 2010, PHMSA published an advance notice of proposed rulemaking (ANPRM; 75 FR...

  19. Materials Reliability Program: Risk-Informed Revision of ASME Section XI Appendix G - Proof of Concept (MRP-143)

    SciTech Connect

    B. Bishop; et al

    2005-03-30

    This study indicates that risk-informed methods can be used to significantly relax the current ASME and NRC Appendix G requirements while still maintaining satisfactory levels of reactor vessel structural integrity. This relaxation in Appendix G requirements directly translates into significant improvements in operational flexibility.

  20. Review of the margins for ASME code fatigue design curve - effects of surface roughness and material variability.

    SciTech Connect

    Chopra, O. K.; Shack, W. J.; Energy Technology

    2003-10-03

    The ASME Boiler and Pressure Vessel Code provides rules for the construction of nuclear power plant components. The Code specifies fatigue design curves for structural materials. However, the effects of light water reactor (LWR) coolant environments are not explicitly addressed by the Code design curves. Existing fatigue strain-vs.-life ({var_epsilon}-N) data illustrate potentially significant effects of LWR coolant environments on the fatigue resistance of pressure vessel and piping steels. This report provides an overview of the existing fatigue {var_epsilon}-N data for carbon and low-alloy steels and wrought and cast austenitic SSs to define the effects of key material, loading, and environmental parameters on the fatigue lives of the steels. Experimental data are presented on the effects of surface roughness on the fatigue life of these steels in air and LWR environments. Statistical models are presented for estimating the fatigue {var_epsilon}-N curves as a function of the material, loading, and environmental parameters. Two methods for incorporating environmental effects into the ASME Code fatigue evaluations are discussed. Data available in the literature have been reviewed to evaluate the conservatism in the existing ASME Code fatigue evaluations. A critical review of the margins for ASME Code fatigue design curves is presented.

  1. 46 CFR 53.01-3 - Adoption of section IV of the ASME Boiler and Pressure Vessel Code.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... reference; see 46 CFR 53.01-1) as limited, modified, or replaced by specific requirements in this part. The... 46 Shipping 2 2013-10-01 2013-10-01 false Adoption of section IV of the ASME Boiler and Pressure...) MARINE ENGINEERING HEATING BOILERS General Requirements § 53.01-3 Adoption of section IV of the...

  2. 46 CFR 53.01-3 - Adoption of section IV of the ASME Boiler and Pressure Vessel Code.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... reference; see 46 CFR 53.01-1) as limited, modified, or replaced by specific requirements in this part. The... 46 Shipping 2 2014-10-01 2014-10-01 false Adoption of section IV of the ASME Boiler and Pressure...) MARINE ENGINEERING HEATING BOILERS General Requirements § 53.01-3 Adoption of section IV of the...

  3. 46 CFR 53.01-3 - Adoption of section IV of the ASME Boiler and Pressure Vessel Code.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... reference; see 46 CFR 53.01-1) as limited, modified, or replaced by specific requirements in this part. The... 46 Shipping 2 2011-10-01 2011-10-01 false Adoption of section IV of the ASME Boiler and Pressure...) MARINE ENGINEERING HEATING BOILERS General Requirements § 53.01-3 Adoption of section IV of the...

  4. 46 CFR 53.01-3 - Adoption of section IV of the ASME Boiler and Pressure Vessel Code.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... reference; see 46 CFR 53.01-1) as limited, modified, or replaced by specific requirements in this part. The... 46 Shipping 2 2010-10-01 2010-10-01 false Adoption of section IV of the ASME Boiler and Pressure...) MARINE ENGINEERING HEATING BOILERS General Requirements § 53.01-3 Adoption of section IV of the...

  5. 46 CFR 53.01-3 - Adoption of section IV of the ASME Boiler and Pressure Vessel Code.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... reference; see 46 CFR 53.01-1) as limited, modified, or replaced by specific requirements in this part. The... 46 Shipping 2 2012-10-01 2012-10-01 false Adoption of section IV of the ASME Boiler and Pressure...) MARINE ENGINEERING HEATING BOILERS General Requirements § 53.01-3 Adoption of section IV of the...

  6. Impact of the A18.1 ASME Standard on Platform Lifts and Stairway Chairlifts on Accessibility and Usability

    ERIC Educational Resources Information Center

    Balmer, David C.

    2010-01-01

    This article summarizes the effect of the ASME A18.1 Standard concerning accessibility and usability of Platform Lifts and their remaining technological challenges. While elevators are currently the most effective means of vertical transportation related to speed, capacity, rise and usability, their major drawbacks for accessibility are cost and…

  7. Verification of Allowable Stresses In ASME Section III Subsection NH For Grade 91 Steel & Alloy 800H

    SciTech Connect

    R. W. Swindeman; M. J. Swindeman; B. W. Roberts; B. E. Thurgood; D. L. Marriott

    2007-11-30

    The database for the creep-rupture of 9Cr-1Mo-V (Grade 91) steel was collected and reviewed to determine if it met the needs for recommending time-dependent strength values, S{sub t}, for coverage in ASME Section III Subsection NH (ASME III-NH) to 650 C (1200 F) and 600,000 hours. The accumulated database included over 300 tests for 1% total strain, nearly 400 tests for tertiary creep, and nearly 1700 tests to rupture. Procedures for analyzing creep and rupture data for ASME III-NH were reviewed and compared to the procedures used to develop the current allowable stress values for Gr 91 for ASME II-D. The criteria in ASME III-NH for estimating S{sub t} included the average strength for 1% total strain for times to 600,000 hours, 80% of the minimum strength for tertiary creep for times to 600,000 hours, and 67% of the minimum rupture strength values for times to 600,000 hours. Time-temperature-stress parametric formulations were selected to correlate the data and make predictions of the long-time strength. It was found that the stress corresponding to 1% total strain and the initiation of tertiary creep were not the controlling criteria over the temperature-time range of concern. It was found that small adjustments to the current values in III-NH could be introduced but that the existing values were conservative and could be retained. The existing database was found to be adequate to extend the coverage to 600,000 hours for temperatures below 650 C (1200 F).

  8. Corticosteroid-Induced MKP-1 Represses Pro-Inflammatory Cytokine Secretion by Enhancing Activity of Tristetraprolin (TTP) in ASM Cells.

    PubMed

    Prabhala, Pavan; Bunge, Kristin; Ge, Qi; Ammit, Alaina J

    2016-10-01

    Exaggerated cytokine secretion drives pathogenesis of a number of chronic inflammatory diseases, including asthma. Anti-inflammatory pharmacotherapies, including corticosteroids, are front-line therapies and although they have proven clinical utility, the molecular mechanisms responsible for their actions are not fully understood. The corticosteroid-inducible gene, mitogen-activated protein kinase (MAPK) phosphatase 1 (MKP-1, DUSP1) has emerged as a key molecule responsible for the repressive effects of steroids. MKP-1 is known to deactivate p38 MAPK phosphorylation and can control the expression and activity of the mRNA destabilizing protein-tristetraprolin (TTP). But whether corticosteroid-induced MKP-1 acts via p38 MAPK-mediated modulation of TTP function in a pivotal airway cell type, airway smooth muscle (ASM), was unknown. While pretreatment of ASM cells with the corticosteroid dexamethasone (preventative protocol) is known to reduce ASM synthetic function in vitro, the impact of adding dexamethasone after stimulation (therapeutic protocol) had not been explored. Whether dexamethasone modulates TTP in a p38 MAPK-dependent manner in this cell type was also unknown. We address this herein and utilize an in vitro model of asthmatic inflammation where ASM cells were stimulated with the pro-asthmatic cytokine tumor necrosis factor (TNF) and the impact of adding dexamethasone 1 h after stimulation assessed. IL-6 mRNA expression and protein secretion was significantly repressed by dexamethasone acting in a temporally distinct manner to increase MKP-1, deactivate p38 MAPK, and modulate TTP phosphorylation status. In this way, dexamethasone-induced MKP-1 acts via p38 MAPK to switch on the mRNA destabilizing function of TTP to repress pro-inflammatory cytokine secretion from ASM cells. J. Cell. Physiol. 231: 2153-2158, 2016. © 2016 Wiley Periodicals, Inc. PMID:26825339

  9. Cyclic-load crack growth in ASME SA-105 grade II steel in high-pressure hydrogen at ambient temperature

    NASA Technical Reports Server (NTRS)

    Walter, R. J.; Chandler, W. T.

    1976-01-01

    ASME SA-105 Grade II steel, which is used in high-pressure hydrogen compressor systems, is similar to steels used or considered for use in high-pressure hydrogen storage vessels and pipelines. This paper summarizes the results of a program conducted to provide cyclic-load crack growth rate (da/dN) data for a fracture mechanics analysis of a 15,000 psi hydrogen compressor facility which contains pulse quieter and after-cooler separator vessels constructed of the ASME SA-105 Grade II steel. Included in the program were tests performed to assist in establishing operating procedures that could minimize the effect of hydrogen on crack growth rates during operation.

  10. Application of the Load Coefficient Method of ASME Code Case N-468 to the seismic analysis of piping systems

    SciTech Connect

    Antaki, G.A.

    1991-01-01

    ASME Code case N-468 recognizes the use of static analysis (the Load Coefficient Method or LCM) as an alternative to the commonly used response spectra modal analysis method (or RSMAM) for the structural evaluation of piping systems. The LCM, in various forms, has been commonly used in the late 1960's to mid-1970's for the design of nuclear piping systems of all sizes. With the advent of more user-friendly software, the LCM slowly gave way to the RSMAM, the latter being almost exclusively used throughout the 1980's. The paper presents the development of the seismic load coefficients in accordance with ASME Section III Code Case N-468. The load coefficients are then applied to 87 piping systems and compared to the response spectra modal analysis method.

  11. Application of the Load Coefficient Method of ASME Code Case N-468 to the seismic analysis of piping systems

    SciTech Connect

    Antaki, G.A.

    1991-12-31

    ASME Code case N-468 recognizes the use of static analysis (the Load Coefficient Method or LCM) as an alternative to the commonly used response spectra modal analysis method (or RSMAM) for the structural evaluation of piping systems. The LCM, in various forms, has been commonly used in the late 1960`s to mid-1970`s for the design of nuclear piping systems of all sizes. With the advent of more user-friendly software, the LCM slowly gave way to the RSMAM, the latter being almost exclusively used throughout the 1980`s. The paper presents the development of the seismic load coefficients in accordance with ASME Section III Code Case N-468. The load coefficients are then applied to 87 piping systems and compared to the response spectra modal analysis method.

  12. Incinerator ash bill supported by ASME (American Society of Mechanical Engineers)

    SciTech Connect

    Not Available

    1989-08-01

    Municipal waste combustion facilities must be retained as an option for solving this nation's total solid waste management problems, John W. Norton, representing the Solid Waste Processing Division of the American Society of Mechanical Engineers (ASME), told the House Subcommittee on Transportation and Hazardous Materials. Norton testified on H.R. 2162, the Municipal Solid Waste Incineration Act of 1989. The bill would ensure environmentally sound disposal of incinerator ash residues while improving public confidence in the disposal of municipal waste through combustion. Although agreeing that the EPA should issue guidelines removing certain items (such as batteries) from the trash to be burned, Norton urged additional research to determine the impact of such actions. He cautioned that toxic components such as heavy metals are pervasive in wastes and, therefore, it is unlikely that removal of a few items will significantly affect the quality of either ash or air emissions. We believe recycling and reuse of municipal solid waste incinerator ash offers great promise to reduce the need for disposal in landfills, testified Norton. He urged that the bill authorize a substantial research program on ash recycling and reuse at EPA.

  13. Unsteady Simulation of an ASME Venturi Flow in a Cross Flow

    NASA Astrophysics Data System (ADS)

    Bonifacio, Jeremy; Rahai, Hamid

    2010-11-01

    Unsteady numerical simulations of an ASME venturi flow into a cross flow were performed. The velocity ratios between the venturi flow and the free stream were 25, 50, and 75%. Two cases of the venturi with and without a tube extension have been investigated. The tube extension length was approximately 4D (here D is the inner diameter of the venturi's outlet), connecting the venturi to the bottom surface of the numerical wind tunnel. A finite volume approach with the Wilcox K-φ turbulence model were used. Results that include contours of the mean velocity, velocity vector, turbulent kinetic energy, pressure and vortices within the venturi as well as downstream in the interaction region indicate that when the venturi is flushed with the surface, there is evidence of flow separation within the venturi, near the outlet. However, when the tube extension was added, the pressure recovery was sustained and flow separation within the venturi was not present and the characteristics of the flow in the interaction region were similar to the corresponding characteristics of a pipe jet in a cross flow.

  14. ASME B89.4.19 Performance Evaluation Tests and Geometric Misalignments in Laser Trackers

    PubMed Central

    Muralikrishnan, B.; Sawyer, D.; Blackburn, C.; Phillips, S.; Borchardt, B.; Estler, W. T.

    2009-01-01

    Small and unintended offsets, tilts, and eccentricity of the mechanical and optical components in laser trackers introduce systematic errors in the measured spherical coordinates (angles and range readings) and possibly in the calculated lengths of reference artifacts. It is desirable that the tests described in the ASME B89.4.19 Standard [1] be sensitive to these geometric misalignments so that any resulting systematic errors are identified during performance evaluation. In this paper, we present some analysis, using error models and numerical simulation, of the sensitivity of the length measurement system tests and two-face system tests in the B89.4.19 Standard to misalignments in laser trackers. We highlight key attributes of the testing strategy adopted in the Standard and propose new length measurement system tests that demonstrate improved sensitivity to some misalignments. Experimental results with a tracker that is not properly error corrected for the effects of the misalignments validate claims regarding the proposed new length tests. PMID:27504211

  15. A case study for assessment of consistency between ASME Section III and XI rules

    SciTech Connect

    Zhao, Y.; Wilkowski, G.

    1996-12-01

    A case study was conducted to evaluate how the current ASME Boiler and Pressure Vessel (B and PV) Code Section XI nuclear piping flaw evaluation rules are affected by the design stress changes in increasing the allowable stress with the B and PV Code Section III piping design rules. A piping model was developed for this purpose from a realistic nuclear piping referred to as BM3 that has been used widely for developing or evaluating the nuclear piping design rules. The piping system was modified so that all the conditions required by the 1995 Section III rules are met. Seismic excitations and internal pressure were increased to reach the maximum allowable stresses under the Section III rules including the effects of pressure, deadweight, seismic inertia, seismic anchor motion, and thermal expansion. The stresses thus calculated elastically using a finite element analysis were then combined using the Section XI rules to evaluate whether the maximum stress ratios for flaw acceptance are exceeded that are allowed by Section XI IWB-3514 for the workmanship standard flaw sizes corresponding to current nondestructive examination capability at butt weld areas. Quantitative results were provided for discussions.

  16. D0 Silicon Upgrade: ASME Code and Pressure Calculations for Liquid Nitrogen Subcooler

    SciTech Connect

    Kuwazaki, Andrew; Leicht, Todd; /Fermilab

    1995-10-04

    Included in this engineering note are three separate calculation divisions. The first calculations are the determination of the required thickness of the LN{sub 2} subcooler flat head according to ASME code. This section includes Appendix A-C. The minimum plate thickness determined was 0.563 in. The actual thickness chosen in fabrication was a 3/4-inch plate milled to 0.594-inch at the bolt circle. Along with the plate thickness, this section calculates the required reinforcement area at the top plate penetrations. It was found that a 1/4-inch fillet weld at each penetration was adequate. The next set of calculations were done to prove that the subcooler internal pressure will always be less than 15 psig and therefore will not be classified as a pressure vessel. The subcooler is always open to a vent pipe. Appendix D calculations show that the vent pipe has a capacity of 1042 lbs/hr if 15 psig is present at the subcooler. It goes on to show that the inlet piping would at that flow rate, see a pressure drop of 104 psig. The maximum supply pressure of the LN{sub 2} storage dewar is 50 psig. Appendix E addresses required flow rates for steady state, loss of vacuum, or fire conditions. Page E9 shows a summary which states the maximum pressure would be 1.50 psig at fire conditions and internal pressure.

  17. Three-dimensional ASM-based segmentation of the subcortical nucleus from volumetric MR images

    NASA Astrophysics Data System (ADS)

    Fu, Yili; Gao, Wenpeng; Xiao, Yongfei; Wang, Shuguo

    2009-10-01

    Delineation of the subcortical nucleus in MR images is prerequisite for advanced radiotheraphy, surgical planning and morphometric analysis. However, it is always difficult to implement such a complicated work. We proposed a novel framework of 3D active shape model (ASM) based segmentation of the subcortical nucleus in MR images. Firstly, the most representative one of all samples represented by the segmented MR volumes is selected as the template and triangulated to generate a triangulated surface mesh. Then, free form deformation is used to establish dense point correspondences between the template and the other samples. A set of consistent triangle meshes are obtained to build the model by a statistical analysis. To fit the model to a MR volume, the model is initialized with Talairach transformation and the edge map around the model is extracted using watershed transform. An algorithm of robust point matching is used to find a transformation matrix and model parameters to transpose the model near the target nucleus and match the model to the target nucleus, respectively. The proposed framework was tested on 18 brain MR volumes. The caudate, putamen, globus pallidus, thalamus, and hippocampus were selected as the objects. In comparison with manual segmentation, the accuracy (Mean+/-SD) of the proposed framework is 0.90+/-0.04 for all objects.

  18. Report No. 2: ASME post-weld heat treating practices: An interpretive report

    SciTech Connect

    Spaeder, C.E. Jr.; Doty, W.D.

    1995-12-01

    The purpose of this report is to review the present ASME Code requirements, identify inconsistencies, provide recommendations for changes and recommend research that should be the basis of further changes in post weld heat treatment (PWHT) practices. Based on a review of the effects of PWHT practices and the present Code rules, it is concluded that present Code rules are generally applicable to high carbon steels produced without notch toughness requirements. For modern-day steels that exhibit high toughness and enhanced weldability, the present Code rules might impose unnecessary requirements and, for certain steels, might actually produce undesirable effects on the properties of the steel. A review of specific requirements related to time and temperature indicate that some are not consistent with literature data and are in need of revision. The authors propose that revisions be made to the Code rules to relate the PWHT requirements to notch toughness and nondestructive examination considerations when a PWHT is not required because of outside considerations (i.e., environmental effects). Such revisions would permit economies in the fabrication of some pressure vessels to take advantage of the effect of high toughness on service performance. Specific research that would be the basis of changes in Code requirements is presented.

  19. The use of lamination analysis and the Tsai-Wu stress criterion in ASME standards for fiber reinforced plastic vessels

    SciTech Connect

    Conlisk, P.J.

    1996-12-01

    The ASME publishes two standards on FRP (Fiber Reinforced Plastic) vessels: Section X of the Boiler and Pressure Vessel Code, and RTP-1 (Reinforced Thermoset Plastic Corrosion Resistant Equipment), which concerns low pressure FRP vessels. The paper describes the application of lamination theory and the Tsai-Wu tensor strength criterion to qualifying designs to the two standards. Numerical examples of application of the theory to typical laminates subjected to pure membrane, pure bending, and combined membrane and bending loads are presented.

  20. High Level Analysis, Design and Validation of Distributed Mobile Systems with CoreASM

    NASA Astrophysics Data System (ADS)

    Farahbod, R.; Glässer, U.; Jackson, P. J.; Vajihollahi, M.

    System design is a creative activity calling for abstract models that facilitate reasoning about the key system attributes (desired requirements and resulting properties) so as to ensure these attributes are properly established prior to actually building a system. We explore here the practical side of using the abstract state machine (ASM) formalism in combination with the CoreASM open source tool environment for high-level design and experimental validation of complex distributed systems. Emphasizing the early phases of the design process, a guiding principle is to support freedom of experimentation by minimizing the need for encoding. CoreASM has been developed and tested building on a broad scope of applications, spanning computational criminology, maritime surveillance and situation analysis. We critically reexamine here the CoreASM project in light of three different application scenarios.

  1. Development and application of proposed ASME Section XI Code changes for risk-based inspection of piping

    SciTech Connect

    West, R.A.

    1996-12-01

    This synopsis has been written to describe a perspective on the development and application of ASME Section XI Code changes for risk-based inspection of piping. The content is specifically related to the use of risk-based technology for Inservice Inspection (ISI) of piping and efforts made to support the ASME Research/Westinghouse Owners Group/Millstone Unit 3 approach for use of this technology. The opinions contained herein may or may not reflect those of the ASME Codes and Standards Committees responsible for these activities. In order to take such a detailed technical subject and put it into an understandable format, the author has chosen to provide an analogy to simplify what is actually taking place. Risk-based technology in the ISI of piping can be likened to the process of making and using specifically ground prescription glasses to allow for better vision. It provides a process to develop and use these uniquely ground glasses that will dynamically focus on all the locations and obstacles within a plant`s piping systems that could cause that plant to trip and fall; more importantly it identifies the locations where the fall could possibly hurt someone else. In this way, Nuclear Safety is being addressed.

  2. Factor Associated with Neutral Sphingomyelinase Activity Mediates Navigational Capacity of Leukocytes Responding to Wounds and Infection: Live Imaging Studies in Zebrafish Larvae

    PubMed Central

    Boecke, Alexandra; Sieger, Dirk; Neacsu, Cristian Dan; Kashkar, Hamid

    2012-01-01

    Factor associated with neutral sphingomyelinase activity (FAN) is an adaptor protein that specifically binds to the p55 receptor for TNF (TNF-RI). Our previous investigations demonstrated that FAN plays a role in TNF-induced actin reorganization by connecting the plasma membrane with actin cytoskeleton, suggesting that FAN may impact on cellular motility in response to TNF and in the context of immune inflammatory conditions. In this study, we used the translucent zebrafish larvae for in vivo analysis of leukocyte migration after morpholino knockdown of FAN. FAN-deficient zebrafish leukocytes were impaired in their migration toward tail fin wounds, leading to a reduced number of cells reaching the wound. Furthermore, FAN-deficient leukocytes show an impaired response to bacterial infections, suggesting that FAN is generally required for the directed chemotactic response of immune cells independent of the nature of the stimulus. Cell-tracking analysis up to 3 h after injury revealed that the reduced number of leukocytes is not due to a reduction in random motility or speed of movement. Leukocytes from FAN-deficient embryos protrude pseudopodia in all directions instead of having one clear leading edge. Our results suggest that FAN-deficient leukocytes exhibit an impaired navigational capacity, leading to a disrupted chemotactic response. PMID:22802420

  3. Imaging the early stages of phospholipase C/sphingomyelinase activity on vesicles containing coexisting ordered-disordered and gel-fluid domains[S

    PubMed Central

    Ibarguren, Maitane; López, David J.; Montes, L.-Ruth; Sot, Jesús; Vasil, Adriana I.; Vasil, Michael L.; Goñi, Félix M.; Alonso, Alicia

    2011-01-01

    The binding and early stages of activity of a phospholipase C/sphingomyelinase from Pseudomonas aeruginosa on giant unilamellar vesicles (GUV) have been monitored using fluorescence confocal microscopy. Both the lipids and the enzyme were labeled with specific fluorescent markers. GUV consisted of a mixture of phosphatidylcholine, sphingomyelin, phosphatidylethanolamine, and cholesterol in equimolar ratios, to which 5–10 mol% of the enzyme end-product ceramide and/or diacylglycerol were occasionally added. Morphological examination of the GUV in the presence of enzyme reveals that, although the enzyme diffuses rapidly throughout the observation chamber, detectable enzyme binding appears to be a slow, random process, with new bound-enzyme-containing vesicles appearing for several minutes. Enzyme binding to the vesicles appears to be a cooperative process. After the initial cluster of bound enzyme is detected, further binding and catalytic activity follow rapidly. After the activity has started, the enzyme is not released by repeated washing, suggesting a “scooting” mechanism for the hydrolytic activity. The enzyme preferentially binds the more disordered domains, and, in most cases, the catalytic activity causes the disordering of the other domains. Simultaneously, peanut- or figure-eight-shaped vesicles containing two separate lipid domains become spherical. At a further stage of lipid hydrolysis, lipid aggregates are formed and vesicles disintegrate. PMID:21252263

  4. SPASM: a 3D-ASM for segmentation of sparse and arbitrarily oriented cardiac MRI data.

    PubMed

    van Assen, Hans C; Danilouchkine, Mikhail G; Frangi, Alejandro F; Ordás, Sebastián; Westenberg, Jos J M; Reiber, Johan H C; Lelieveldt, Boudewijn P F

    2006-04-01

    A new technique (SPASM) based on a 3D-ASM is presented for automatic segmentation of cardiac MRI image data sets consisting of multiple planes with arbitrary orientations, and with large undersampled regions. Model landmark positions are updated in a two-stage iterative process. First, landmark positions close to intersections with images are updated. Second, the update information is propagated to the regions without image information, such that new locations for the whole set of the model landmarks are obtained. Feature point detection is performed by a fuzzy inference system, based on fuzzy C-means clustering. Model parameters were optimized on a computer cluster and the computational load distributed by grid computing. SPASM was applied to image data sets with an increasing sparsity (from 2 to 11 slices) comprising images with different orientations and stemming from different MRI acquisition protocols. Segmentation outcomes and calculated volumes were compared to manual segmentation on a dense short-axis data configuration in a 3D manner. For all data configurations, (sub-)pixel accuracy was achieved. Performance differences between data configurations were significantly different (p<0.05) for SA data sets with less than 6 slices, but not clinically relevant (volume differences<4 ml). Comparison to results from other 3D model-based methods showed that SPASM performs comparable to or better than these other methods, but SPASM uses considerably less image data. Sensitivity to initial model placement proved to be limited within a range of position perturbations of approximately 20 mm in all directions. PMID:16439182

  5. Towards a consensus-based biokinetic model for green microalgae - The ASM-A.

    PubMed

    Wágner, Dorottya S; Valverde-Pérez, Borja; Sæbø, Mariann; Bregua de la Sotilla, Marta; Van Wagenen, Jonathan; Smets, Barth F; Plósz, Benedek Gy

    2016-10-15

    Cultivation of microalgae in open ponds and closed photobioreactors (PBRs) using wastewater resources offers an opportunity for biochemical nutrient recovery. Effective reactor system design and process control of PBRs requires process models. Several models with different complexities have been developed to predict microalgal growth. However, none of these models can effectively describe all the relevant processes when microalgal growth is coupled with nutrient removal and recovery from wastewaters. Here, we present a mathematical model developed to simulate green microalgal growth (ASM-A) using the systematic approach of the activated sludge modelling (ASM) framework. The process model - identified based on a literature review and using new experimental data - accounts for factors influencing photoautotrophic and heterotrophic microalgal growth, nutrient uptake and storage (i.e. Droop model) and decay of microalgae. Model parameters were estimated using laboratory-scale batch and sequenced batch experiments using the novel Latin Hypercube Sampling based Simplex (LHSS) method. The model was evaluated using independent data obtained in a 24-L PBR operated in sequenced batch mode. Identifiability of the model was assessed. The model can effectively describe microalgal biomass growth, ammonia and phosphate concentrations as well as the phosphorus storage using a set of average parameter values estimated with the experimental data. A statistical analysis of simulation and measured data suggests that culture history and substrate availability can introduce significant variability on parameter values for predicting the reaction rates for bulk nitrate and the intracellularly stored nitrogen state-variables, thereby requiring scenario specific model calibration. ASM-A was identified using standard cultivation medium and it can provide a platform for extensions accounting for factors influencing algal growth and nutrient storage using wastewater resources. PMID:27525381

  6. A spontaneous genomic deletion in Listeria ivanovii identifies LIPI-2, a species-specific pathogenicity island encoding sphingomyelinase and numerous internalins.

    PubMed

    Domínguez-Bernal, Gustavo; Müller-Altrock, Stefanie; González-Zorn, Bruno; Scortti, Mariela; Herrmann, Petra; Monzó, Héctor J; Lacharme, Lizeth; Kreft, Jürgen; Vázquez-Boland, José A

    2006-01-01

    Listeria ivanovii differs from the human pathogen Listeria monocytogenes in that it specifically affects ruminants, causing septicaemia and abortion but not meningo-encephalitis. The genetic characterization of spontaneous L. ivanovii mutants lacking the virulence factor SmcL (sphingomyelinase) led us to identify LIPI-2, the first species-specific pathogenicity island from Listeria. Besides SmcL, this 22 kb chromosomal locus encodes 10 internalin (Inl) proteins: i-InlB1 and -B2 are large/surface-associated Inls similar to L. monocytogenes InlB; i-InlE to -L are small/excreted (SE)-Inls, i-InlG being a tandem fusion of two SE-Inls. Except i-inlB1, all LIPI-2 inl genes are controlled by the virulence regulator, PrfA. LIPI-2 is inserted into a tRNA locus and is unstable - half of it deleting at approximately 10(-4) frequency with a portion of contiguous DNA. The spontaneous mutants were attenuated in vivo in mice and lambs and showed impaired intracellular growth and apoptosis induction in bovine MDBK cells. Targeted knock-out mutations associated the virulence defect with LIPI-2 genes. The region between the core genome loci ysnB-tRNA(arg) and ydeI flanking LIPI-2 contained different gene complements in the different Listeria spp. and even serovars of L. monocytogenes, including remnants of the PSA bacteriophage int gene in serovar 4b, indicating it is a hot spot for horizontal genome diversification. LIPI-2 is conserved in L. ivanovii ssp. ivanovii and londoniensis, suggesting an early acquisition during the species' evolution. LIPI-2 is likely to play an important role in the pathogenic and host tropism of L. ivanovii. PMID:16390439

  7. Alkaline sphingomyelinase (NPP7) promotes cholesterol absorption by affecting sphingomyelin levels in the gut: A study with NPP7 knockout mice.

    PubMed

    Zhang, Ping; Chen, Ying; Cheng, Yajun; Hertervig, Erik; Ohlsson, Lena; Nilsson, Ake; Duan, Rui-Dong

    2014-05-15

    We previously showed that dietary sphingomyelin (SM) inhibited cholesterol absorption in animals. The key enzyme hydrolyzing SM in the gut is alkaline sphingomyelinase (alk-SMase, nucleotide pyrophosphatase/phosphodiesterase 7). Here using the fecal dual-isotope ratio method we compared cholesterol absorption in the wild-type (WT) and alk-SMase knockout (KO) mice. The animals were fed an emulsion containing [(14)C]cholesterol and [(3)H]sitosterol. The radioactivities in the lipids of the fecal samples collected 4, 8, and 24 h thereafter were determined, and the ratio of (14)C/(3)H was calculated. We found that the fecal [(14)C]cholesterol recovery in the KO mice was significantly higher than in the WT mice. A maximal 92% increase occurred 8 h after feeding. Recovery of [(3)H]sitosterol did not differ between the two groups. Accordingly, the (14)C-to-(3)H ratio of fecal lipids was 133% higher at 8 h and 75% higher at 24 h in the KO than in the WT mice. Decreased [(14)C]cholesterol was also found in the serum of the KO mice 4 h after feeding. Supplement of SM in the emulsion reduced the differences in fecal [(14)C]cholesterol recovery between the WT and KO mice because of a greater increase of [(14)C]cholesterol recovery in the WT mice. Without treatment, the KO mice had significantly higher SM levels in the intestinal content and feces, but not in the intestinal mucosa or serum. The expression of Niemann-Pick C1 like 1 protein in the small intestine was not changed. In conclusion, alk-SMase is a physiological factor promoting cholesterol absorption by reducing SM levels in the intestinal lumen. PMID:24650549

  8. Bone Morphogenic Protein (BMP) Signaling Up-regulates Neutral Sphingomyelinase 2 to Suppress Chondrocyte Maturation via the Akt Protein Signaling Pathway as a Negative Feedback Mechanism*

    PubMed Central

    Kakoi, Hironori; Maeda, Shingo; Shinohara, Naohiro; Matsuyama, Kanehiro; Imamura, Katsuyuki; Kawamura, Ichiro; Nagano, Satoshi; Setoguchi, Takao; Yokouchi, Masahiro; Ishidou, Yasuhiro; Komiya, Setsuro

    2014-01-01

    Although bone morphogenic protein (BMP) signaling promotes chondrogenesis, it is not clear whether BMP-induced chondrocyte maturation is cell-autonomously terminated. Loss of function of Smpd3 in mice results in an increase in mature hypertrophic chondrocytes. Here, we report that in chondrocytes the Runx2-dependent expression of Smpd3 was increased by BMP-2 stimulation. Neutral sphingomyelinase 2 (nSMase2), encoded by the Smpd3 gene, was detected both in prehypertrophic and hypertrophic chondrocytes of mouse embryo bone cartilage. An siRNA for Smpd3, as well as the nSMase inhibitor GW4869, significantly enhanced BMP-2-induced differentiation and maturation of chondrocytes. Conversely, overexpression of Smpd3 or C2-ceramide, which mimics the function of nSMase2, inhibited chondrogenesis. Upon induction of Smpd3 siRNA or GW4869, phosphorylation of both Akt and S6 proteins was increased. The accelerated chondrogenesis induced by Smpd3 silencing was negated by application of the Akt inhibitor MK2206 or the mammalian target of rapamycin inhibitor rapamycin. Importantly, in mouse bone culture, GW4869 treatment significantly promoted BMP-2-induced hypertrophic maturation and calcification of chondrocytes, which subsequently was eliminated by C2-ceramide. Smpd3 knockdown decreased the apoptosis of terminally matured ATDC5 chondrocytes, probably as a result of decreased ceramide production. In addition, we found that expression of hyaluronan synthase 2 (Has2) was elevated by a loss of Smpd3, which was restored by MK2206. Indeed, expression of Has2 protein decreased in nSMase2-positive hypertrophic chondrocytes in the bones of mouse embryos. Our data suggest that the Smpd3/nSMase2-ceramide-Akt signaling axis negatively regulates BMP-induced chondrocyte maturation and Has2 expression to control the rate of endochondral ossification as a negative feedback mechanism. PMID:24505141

  9. Impact of the A18.1 ASME Standard on platform lifts and stairway chairlifts on accessibility and usability.

    PubMed

    Balmer, David C

    2010-01-01

    ABSTRACT This article summarizes the effect of the ASME A18.1 Standard concerning accessibility and usability of Platform Lifts and their remaining technological challenges. While elevators are currently the most effective means of vertical transportation related to speed, capacity, rise and usability, their major drawbacks for accessibility are cost and space. Platform lifts and stairway chairlifts remain the "devices of choice" for small elevation changes in existing buildings. ADAAG limits them to very specific circumstances in new construction. The ASME A18.1 Standard addresses the safety requirements of inclined stairway chairlifts (which are not ADA compliant) and inclined and vertical platform lifts (which are ADA Compliant). Chairlifts do not provide access for wheeled mobility devices. Restricting access by means of keys is eliminated, inclined platform lift designs that do not interfere with stairway traffic, promoting new ideas for the design of vertical lifts, increasing the allowable vertical travel of a lift and strengthening lift ramps to improve their safety. Despite design advances inherent in the A18.1, significant platform lift usability issues continue to exist. Increased sizes and weights of powered mobility devices indicate that the permitted lift platform area be modified and that permitted weight capacities be codified as minimums instead of maximums. PMID:20402046

  10. ASME AG-1 REQUIREMENT EXEMPTION JUSTIFICATIONS FOR VENTILATION SYSTEMS AT NUCLEAR WASTE STORAGE TANKS AT THE HANFORD SITE

    SciTech Connect

    GUSTAVSON, R.D.

    2004-09-03

    Washington State Department of Health regulations require compliance with the American Society of Mechanical Engineers (ASME) AG-1, ''Code on Nuclear Air and Gas Treatment,'' for all new radioactive air emission units. As a result, these requirements have been applied to systems that ventilate the radioactive waste storage tanks in the tank farm facilities on the U.S. Department of Energy's Hanford Site. ASME AG-1 is applied as a regulatory constraint to waste tank ventilation systems at the Hanford Site, even though the code was not intended for these systems. An assessment was performed to identify which requirements should be exempted for waste tank ventilation systems. The technical justifications for requirement exemptions were prepared and presented to the regulator. The technical justifications were documented so that select requirement exemptions for specific projects and systems can be sought through the regulator's permitting process. This paper presents the rationale for attempting to receive requirement exemption and presents examples of the technical justifications that form the basis for these exemptions.

  11. Modification of ASM3 for the determination of biomass adsorption/storage capacity in bulking sludge control.

    PubMed

    Makinia, J; Rosenwinkel, K H; Phan, L C

    2006-01-01

    The selector activated sludge (SAS) systems are known to prevent excessive growth of filamentous microorganisms responsible for bulking sludge, but these systems were hardly ever modelled. This study aimed to develop a model capable of predicting rapid substrate removal in the SAS systems. For this purpose, the Activated Sludge Model No. 3 (ASM3) was extended with three processes (adsorption, direct growth on the adsorbed substrate under aerobic or anoxic conditions). The modified ASM3 was tested against the results of batch experiments with the biomass originating from two full-scale SAS systems in Germany. The endogenous biomass was mixed with various readily biodegradable substrates (acetate, peptone, glucose and wastewater) and the utilisation of substrate (expresses as COD) and oxygen uptake rates (OURs) were measured during the experiments. In general, model predictions fitted to the experimental data, but a considerable number of kinetic (5) and stoichiometric (2) parameters needed to be adjusted during model calibration. The simulation results revealed that storage was generally a dominating process compared to direct growth in terms of the adsorbed substrate utilisation. The contribution of storage ranged from 65-71% (Plant A) and 69-92% (Plant B). PMID:16605021

  12. TECHNICAL BASIS AND APPLICATION OF NEW RULES ON FRACTURE CONTROL OF HIGH PRESSURE HYDROGEN VESSEL IN ASME SECTION VIII, DIVISION 3 CODE

    SciTech Connect

    Rawls, G

    2007-04-30

    As a part of an ongoing activity to develop ASME Code rules for the hydrogen infrastructure, the ASME Boiler and Pressure Vessel Code Committee approved new fracture control rules for Section VIII, Division 3 vessels in 2006. These rules have been incorporated into new Article KD-10 in Division 3. The new rules require determining fatigue crack growth rate and fracture resistance properties of materials in high pressure hydrogen gas. Test methods have been specified to measure these fracture properties, which are required to be used in establishing the vessel fatigue life. An example has been given to demonstrate the application of these new rules.

  13. Protective antibodies against a sphingomyelinase D from Loxosceles intermedia spider venom elicited in mice with different genetic background.

    PubMed

    Oliveira, Camila Franco Batista; Vilela, Andrea; Coura, Luis Augusto M; Rodrigues, Fernandes Tenório Gomes; Nagem, Ronaldo Alves Pinto; Chávez-Olortegui, Carlos; Maioli, Tatiani U; Felicori, Liza F

    2016-07-19

    In the present investigation we used a recombinant LiD1 toxin, named rLiD1his, from Loxosceles intermedia brown spider to elicit specific antibodies in mice carrying different Human Leukocyte Antigens class II (HLAII) {DRB1.0401 (DR4), DQB1.0601 (DQ6) and DQB1.0302 (DQ8)} as well as in BALB/C and C57BL/6 control mice. All mice strains produced high antibody titers against rLiD1his but DR4 mice antibodies (the lower responder mice) were not able to recognize L. intermedia crude venom. The anti-rLiD1his sera, except from DR4 mice, were able to neutralize dermonecrotic, hemorrhagic and edematogenic effects of rLiD1his in naïve rabbits. Overlapping peptides from the amino acid sequence of LiD1 toxin were prepared by SPOT method and differences in LiD1 epitope recognition were observed using different mice anti-rLiD1his sera. The region (160)DKVGHDFSGNDDISDVGK(177) was recognized by transgenic DQ8 and DQ6 mice sera. Other epitopes were recognized by at least two different animals' sera including (10)MGHMVNAIGQIDEFVNLG(27), (37)FDDNANPEYTYHGIP(51), (70)GLRSATTPGNSKYQEKLV(87) and (259)AAYKKKFRVATYDDN(273). Among these epitopes, the epitopes 37-51 and 160-177 have already been shown in previously studies as good candidates to be used alone or combined with other peptides to induce protective immune response against Loxosceles venoms. The results presented here highlight the importance of HLAII in antibody response and recognition of specific B-cell epitopes of rLiD1his spider toxin according to HLAII type and impact in the epitopic vaccine development against this spider. PMID:27265457

  14. Diacylglycerol-containing docosahexaenoic acid in acyl chain modulates airway smooth muscle tone.

    PubMed

    Hichami, Aziz; Morin, Caroline; Rousseau, Eric; Khan, Naim A

    2005-10-01

    We synthesized and assessed the role of a diacylglycerol (DAG)-containing docosahexaenoic acid (DHA), that is, 1-stearoyl-2-docosahexaenoyl-sn-glycerol (SDHG), in the contraction of guinea pig airway smooth muscle (ASM). We compared its action with 1-stearoyl-2-arachidonoyl-sn-glycerol (SAG) and 1,2-dioctanoyl-sn-glycerol (1,2-DiC8), a stable DAG analog. The three DAGs (SAG, SDHG, and 1,2-DiC8) induced reversible concentration-dependent contraction of ASM. SDHG induced higher guinea pig ASM contraction than did SAG and 1,2-DiC8. The effects of SDHG were blocked, to different extents, by nifedipine (L-type Ca2+ channel blocker). By employing GF-109203X (protein kinase C [PKC] inhibitor) and lanthanum (La3+), a nonselective cation channel blocker, we observed that SDHG evoked ASM contractile response via PKC-dependent and PKC-independent (but Ca2+-dependent) pathways. Interestingly, SAG exerted its action only by increasing [Ca2+]i and did not require PKC activation. To probe the implication of calcium mobilization, we employed thapsigargin (TG), which also induced ASM contraction in a calcium-dependent manner. SDHG and 1,2-DiC8, in a PKC-dependent manner, induced the phosphorylation of CPI-17 (myosin light chain phosphatase inhibitor of 17 kD). Furthermore, SAG and TG failed to phosphorylate CPI-17 in ASM cells. Our results suggest that different DAG species, produced during a dietary supplementation with fatty acids, could modulate the reactivity of airway smooth muscles in a PKC-dependent and -independent manner, and hence, may play a critical role in health and disease. PMID:15961724

  15. Targeting the γ-Aminobutyric Acid A Receptor α4 Subunit in Airway Smooth Muscle to Alleviate Bronchoconstriction.

    PubMed

    Yocum, Gene T; Gallos, George; Zhang, Yi; Jahan, Rajwana; Stephen, Michael Rajesh; Varagic, Zdravko; Puthenkalam, Roshan; Ernst, Margot; Cook, James M; Emala, Charles W

    2016-04-01

    We previously demonstrated that airway smooth muscle (ASM) cells express γ-aminobutyric acid A receptors (GABAARs), and that GABAAR agonists acutely relax ASM. Among the GABAAR α subunits, human ASM cells express only α4 and α5, providing the opportunity for selective pharmacologic targeting. Novel GABAAR-positive allosteric modulators designed for enhanced α4/α6 subunit selectivity were synthesized using iterative computational analyses (CMD-45 and XHe-III-74). Studies using oocyte heterologous expression systems confirmed that CMD-45 and XHe-III-74 led to significantly greater augmentation of currents induced by a 3% maximal effective concentration (EC3) of GABA [EC3]-induced currents in oocytes expressing α4 or α6 subunits (along with β3 and γ2) compared with other α subunits. CMD-45 and XHe-III-74 also led to greater ex vivo relaxation of contracted wild-type mouse tracheal rings compared with tracheal rings from GABAAR α4 subunit (Gabra4) knockout mice. Furthermore, CMD-45 and XHe-III-74 significantly relaxed precontracted human ASM ex vivo, and, at a low concentration, both ligands led to a significant leftward shift in albuterol-mediated ASM relaxation. In vivo, inhaled XHe-III-74 reduced respiratory system resistance in an asthmatic mouse model. Pretreatment of human ASM cells with CMD-45 and XHe-III-74 inhibited histamine-induced increases in intracellular calcium concentrations in vitro, an effect that was lost when calcium was omitted from the extracellular buffer, suggesting that inhibition of calcium influx due to alterations in plasma membrane potential may play a role in the mechanism of ASM relaxation. Selective targeting of the GABAAR α4 subunit with inhaled ligands may be a novel therapeutic pathway to treat bronchoconstriction, while avoiding sedative central nervous system effects, which are largely mediated by α1-3 subunit-containing GABAARs in the brain. PMID:26405827

  16. Systolic and diastolic assessment by 3D-ASM segmentation of gated-SPECT Studies: a comparison with MRI

    NASA Astrophysics Data System (ADS)

    Tobon-Gomez, C.; Bijnens, B. H.; Huguet, M.; Sukno, F.; Moragas, G.; Frangi, A. F.

    2009-02-01

    Gated single photon emission tomography (gSPECT) is a well-established technique used routinely in clinical practice. It can be employed to evaluate global left ventricular (LV) function of a patient. The purpose of this study is to assess LV systolic and diastolic function from gSPECT datasets in comparison with cardiac magnetic resonance imaging (CMR) measurements. This is achieved by applying our recently implemented 3D active shape model (3D-ASM) segmentation approach for gSPECT studies. This methodology allows for generation of 3D LV meshes for all cardiac phases, providing volume time curves and filling rate curves. Both systolic and diastolic functional parameters can be derived from these curves for an assessment of patient condition even at early stages of LV dysfunction. Agreement of functional parameters, with respect to CMR measurements, were analyzed by means of Bland-Altman plots. The analysis included subjects presenting either LV hypertrophy, dilation or myocardial infarction.

  17. Evaluation of the capacity of welded attachments to elbows as compared to the methodology of ASME Code Case N-318

    NASA Astrophysics Data System (ADS)

    Rawls, G. B.; Wais, E. A.; Rodabaugh, E. C.

    This paper presents the results of a series of tests conducted to assess the capacity of various configurations of integral welded attachments. These tests are unique in that the attachments are welded to the outer radius of pipe elbows. The lug configurations tested include both rectangular and cross (cruciform) shapes. Both limit load and fatigue tests are performed on the lug-elbow configurations. The results of the limit load tests are presented as limit moments. The results of the fatigue tests are cycles-to-failure. Markl's equation is then used, with the fatigue results, to determine stress intensification factors. The limit moments and stress intensification factors are then compared to those developed using the methodology of ASME Code Case N-318. The level of conservatism in the Code Case methodology is then compared to the test results.

  18. A guide for the ASME code for austenitic stainless steel containment vessels for high-level radioactive materials

    SciTech Connect

    Raske, D.T.

    1995-06-01

    The design and fabrication criteria recommended by the US Department of Energy (DOE) for high-level radioactive materials containment vessels used in packaging is found in Section III, Division 1, Subsection NB of the ASME Boiler and Pressure Vessel Code. This Code provides material, design, fabrication, examination, and testing specifications for nuclear power plant components. However, many of the requirements listed in the Code are not applicable to containment vessels made from austenitic stainless steel with austenitic or ferritic steel bolting. Most packaging designers, engineers, and fabricators are intimidated by the sheer volume of requirements contained in the Code; consequently, the Code is not always followed and many requirements that do apply are often overlooked during preparation of the Safety Analysis Report for Packaging (SARP) that constitutes the basis to evaluate the packaging for certification.

  19. Comparisons of ANS, ASME, AWS, and NFPA standards cited in the NRC standard review plan, NUREG-0800, and related documents

    SciTech Connect

    Ankrum, A.R.; Bohlander, K.L.; Gilbert, E.R.; Spiesman, J.B.

    1995-11-01

    This report provides the results of comparisons of the cited and latest versions of ANS, ASME, AWS and NFPA standards cited in the NRC Standard Review Plan for the Review of Safety Analysis Reports for Nuclear Power Plants (NUREG 0800) and related documents. The comparisons were performed by Battelle Pacific Northwest Laboratories in support of the NRC`s Standard Review Plan Update and Development Program. Significant changes to the standards, from the cited version to the latest version, are described and discussed in a tabular format for each standard. Recommendations for updating each citation in the Standard Review Plan are presented. Technical considerations and suggested changes are included for related regulatory documents (i.e., Regulatory Guides and the Code of Federal Regulations) citing the standard. The results and recommendations presented in this document have not been subjected to NRC staff review.

  20. Structural diversity in gallium(III) complexes of the tripodal triarsine MeC(CH2AsMe2)3.

    PubMed

    Cheng, Fei; Hector, Andrew L; Levason, William; Reid, Gillian; Webster, Michael; Zhang, Wenjian

    2007-06-01

    The preparation and crystal structures of the first examples of gallium halide complexes with the tripodal arsine, MeC(CH(2)AsMe(2))3, reveal three distinctly different coordination modes for the ligand; the neutral [{micro(3)-MeC(CH(2)AsMe(2))3-kappaAs:kappaAs':kappaAs''}(GaI(3))3] with the triarsine coordinating to three GaI(3) units, [{Me(2)AsCH(2)C(Me)(CH(2)AsMe(2))2-kappa(2)AsAs'}GaCl(2)][GaCl(4)] involving bidentate chelation to a GaCl2+ cationic unit with the third As donor atom uncoordinated, and [{MeC(CH(2)AsMe(2))3-kappaAs:kappa(2)As'As''}(GaCl3)(GaCl2)][GaCl4] in which the triarsine forms a bidentate chelate to the GaCl2+ unit and the third As donor atom binds to a further GaCl3 unit. PMID:17514342

  1. 46 CFR 54.01-2 - Adoption of division 1 of section VIII of the ASME Boiler and Pressure Vessel Code.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... Code (incorporated by reference, see 46 CFR 54.01-1), as limited, modified, or replaced by specific... Boiler and Pressure Vessel Code. 54.01-2 Section 54.01-2 Shipping COAST GUARD, DEPARTMENT OF HOMELAND... division 1 of section VIII of the ASME Boiler and Pressure Vessel Code. (a) Pressure vessels shall...

  2. 46 CFR 54.01-2 - Adoption of division 1 of section VIII of the ASME Boiler and Pressure Vessel Code.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... Code (incorporated by reference, see 46 CFR 54.01-1), as limited, modified, or replaced by specific... Boiler and Pressure Vessel Code. 54.01-2 Section 54.01-2 Shipping COAST GUARD, DEPARTMENT OF HOMELAND... division 1 of section VIII of the ASME Boiler and Pressure Vessel Code. (a) Pressure vessels shall...

  3. 46 CFR 54.01-2 - Adoption of division 1 of section VIII of the ASME Boiler and Pressure Vessel Code.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... Code (incorporated by reference, see 46 CFR 54.01-1), as limited, modified, or replaced by specific... Boiler and Pressure Vessel Code. 54.01-2 Section 54.01-2 Shipping COAST GUARD, DEPARTMENT OF HOMELAND... division 1 of section VIII of the ASME Boiler and Pressure Vessel Code. (a) Pressure vessels shall...

  4. 46 CFR 54.01-2 - Adoption of division 1 of section VIII of the ASME Boiler and Pressure Vessel Code.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... Code (incorporated by reference, see 46 CFR 54.01-1), as limited, modified, or replaced by specific... Boiler and Pressure Vessel Code. 54.01-2 Section 54.01-2 Shipping COAST GUARD, DEPARTMENT OF HOMELAND... division 1 of section VIII of the ASME Boiler and Pressure Vessel Code. (a) Pressure vessels shall...

  5. 46 CFR 54.01-2 - Adoption of division 1 of section VIII of the ASME Boiler and Pressure Vessel Code.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... Code (incorporated by reference, see 46 CFR 54.01-1), as limited, modified, or replaced by specific... Boiler and Pressure Vessel Code. 54.01-2 Section 54.01-2 Shipping COAST GUARD, DEPARTMENT OF HOMELAND... division 1 of section VIII of the ASME Boiler and Pressure Vessel Code. (a) Pressure vessels shall...

  6. Regulatory Safety Issues in the Structural Design Criteria of ASME Section III Subsection NH and for Very High Temperatures for VHTR & GEN IV

    SciTech Connect

    William J. O’Donnell; Donald S. Griffin

    2007-05-07

    The objective of this task is to identify issues relevant to ASME Section III, Subsection NH [1], and related Code Cases that must be resolved for licensing purposes for VHTGRs (Very High Temperature Gas Reactor concepts such as those of PBMR, Areva, and GA); and to identify the material models, design criteria, and analysis methods that need to be added to the ASME Code to cover the unresolved safety issues. Subsection NH was originally developed to provide structural design criteria and limits for elevated-temperature design of Liquid Metal Fast Breeder Reactor (LMFBR) systems and some gas-cooled systems. The U.S. Nuclear Regulatory Commission (NRC) and its Advisory Committee for Reactor Safeguards (ACRS) reviewed the design limits and procedures in the process of reviewing the Clinch River Breeder Reactor (CRBR) for a construction permit in the late 1970s and early 1980s, and identified issues that needed resolution. In the years since then, the NRC and various contractors have evaluated the applicability of the ASME Code and Code Cases to high-temperature reactor designs such as the VHTGRs, and identified issues that need to be resolved to provide a regulatory basis for licensing. This Report describes: (1) NRC and ACRS safety concerns raised during the licensing process of CRBR , (2) how some of these issues are addressed by the current Subsection NH of the ASME Code; and (3) the material models, design criteria, and analysis methods that need to be added to the ASME Code and Code Cases to cover unresolved regulatory issues for very high temperature service.

  7. Assays for in vitro monitoring of proliferation of human airway smooth muscle (ASM) and human pulmonary arterial vascular smooth muscle (VSM) cells.

    PubMed

    Goncharova, Elena A; Lim, Poay; Goncharov, Dmitry A; Eszterhas, Andrew; Panettieri, Reynold A; Krymskaya, Vera P

    2006-01-01

    Vascular and airway remodeling, which are characterized by airway smooth muscle (ASM) and pulmonary arterial vascular smooth muscle (VSM) proliferation, contribute to the pathology of asthma, pulmonary hypertension, restenosis and atherosclerosis. To evaluate the proliferation of VSM and ASM cells in response to mitogens, we perform a [3H]thymidine incorporation assay. The proliferation protocol takes approximately 48 h and includes stimulating cells synchronized in G0/G1 phase of the cell cycle with agonists, labeling cells with [3H]thymidine and examining levels of [3H]thymidine incorporation by scintillation counting. Although using radiolabeled [3H]thymidine incorporation is a limitation, the greatest benefit of the assay is providing reliable and statistically significant data. PMID:17406550

  8. Implementation of ANSI/ASME NQA-1 for development of GA-4 and GA-9 spent fuel casks

    SciTech Connect

    Dunlap, M.G.

    1989-06-01

    The Department of Energy (DOE) Office of Civilian Radioactive Waste Management (OCRWM) has awarded General Atomics (GA) a contract to develop a legal weight truck (LWT) transportation system to transport boiling-water-reactor (BWR) and pressurized water reactor (PWR) spent fuels. DOE's contract requires GA to establish, maintain, and implement an NRC-approved Quality Assurance Program in accordance with ANSI/ASME NQA-1, ''Quality Assurance Program Requirements'' 1986 Edition (basic requirements and supplements), and the 1986 la Addenda. GA's Quality Assurance Program is a company-wide system described in GA's Quality Assurance Manual (QAM). General Atomics' Quality Assurance Program is implemented for a specific project by means of a Quality Assurance Program Document (QAPD). The QAPD is the document used by GA to invoke and document selected quality and project-applicable requirements. The QAPD is a tailored technical quality plan for a specific contract which can add to, revise, or delete certain portions of the basic GA QA Manual. The QAPD then becomes the controlling QA document for a specific project, and the provisions of the QAPD are followed by all project team members as though it were the QA Manual. For the legal weight truck (LWT) transportation project, GA has developed a QAPD that invokes the NRC-approved QA Program and also addresses the additional requirements that are included in the NQA-1 1986 Edition and la Addenda and the additional DOE/ID imposed requirements. 8 refs., 1 fig., 1 tab.

  9. DEVELOPMENT OF ASME SECTION X CODE RULES FOR HIGH PRESSURE COMPOSITE HYDROGEN PRESSURE VESSELS WITH NON-LOAD SHARING LINERS

    SciTech Connect

    Rawls, G.; Newhouse, N.; Rana, M.; Shelley, B.; Gorman, M.

    2010-04-13

    The Boiler and Pressure Vessel Project Team on Hydrogen Tanks was formed in 2004 to develop Code rules to address the various needs that had been identified for the design and construction of up to 15000 psi hydrogen storage vessel. One of these needs was the development of Code rules for high pressure composite vessels with non-load sharing liners for stationary applications. In 2009, ASME approved new Appendix 8, for Section X Code which contains the rules for these vessels. These vessels are designated as Class III vessels with design pressure ranging from 20.7 MPa (3,000 ps)i to 103.4 MPa (15,000 psi) and maximum allowable outside liner diameter of 2.54 m (100 inches). The maximum design life of these vessels is limited to 20 years. Design, fabrication, and examination requirements have been specified, included Acoustic Emission testing at time of manufacture. The Code rules include the design qualification testing of prototype vessels. Qualification includes proof, expansion, burst, cyclic fatigue, creep, flaw, permeability, torque, penetration, and environmental testing.

  10. Elastic-plastic analysis of the PVRC burst disk tests with comparison to the ASME code -- Primary stress limits

    SciTech Connect

    Jones, D.P.; Holliday, J.E.

    1999-02-01

    This paper provides a comparison between finite element analysis results and test data from the Pressure Vessel Research Council (PVRC) burst disk program. Testing sponsored by the PVRC over 20 years ago was done by pressurizing circular flat disks made from three different materials until failure by bursting. The purpose of this re-analysis is to investigate the use of finite element analysis (FEA) to assess the primary stress limits of the ASME Boiler and Pressure Vessel Code (1998) and to qualify the use of elastic-plastic (EP-FEA) for limit load calculations. The three materials tested represent the range of strength and ductility found in modern pressure vessel construction and include a low strength high ductility material, a medium strength medium ductility material, and a high strength low ductility low alloy material. Results of elastic and EP-FEA are compared to test data. Stresses from the elastic analyses are linearized for comparison of Code primary stress limits to test results. Elastic-plastic analyses are done using both best-estimate and elastic-perfectly plastic (EPP) stress-strain curves. Both large strain-large displacement (LSLD) and small strain-small displacement (SSSD) assumptions are used with the EP-FEA. Analysis results are compared to test results to evaluate the various analysis methods, models, and assumptions as applied to the bursting of thin disks.

  11. Comparative study of diverse model building strategies for 3D-ASM segmentation of dynamic gated SPECT data

    NASA Astrophysics Data System (ADS)

    Tobon-Gomez, C.; Butakoff, C.; Ordas, S.; Aguade, S.; Frangi, A. F.

    2007-03-01

    Over the course of the last two decades, myocardial perfusion with Single Photon Emission Computed Tomography (SPECT) has emerged as an established and well-validated method for assessing myocardial ischemia, viability, and function. Gated-SPECT imaging integrates traditional perfusion information along with global left ventricular function. Despite of these advantages, inherent limitations of SPECT imaging yield a challenging segmentation problem, since an error of only one voxel along the chamber surface may generate a huge difference in volume calculation. In previous works we implemented a 3-D statistical model-based algorithm for Left Ventricle (LV) segmentation of in dynamic perfusion SPECT studies. The present work evaluates the relevance of training a different Active Shape Model (ASM) for each frame of the gated SPECT imaging acquisition in terms of their subsequent segmentation accuracy. Models are subsequently employed to segment the LV cavity of gated SPECT studies of a virtual population. The evaluation is accomplished by comparing point-to-surface (P2S) and volume errors, both against a proper Gold Standard. The dataset comprised 40 voxel phantoms (NCAT, Johns Hopkins, University of of North Carolina). Monte-Carlo simulations were generated with SIMIND (Lund University) and reconstructed to tomographic slices with ASPIRE (University of Michigan).

  12. Analysis of systematic errors of the ASM/RXTE monitor and GT-48 γ-ray telescope

    NASA Astrophysics Data System (ADS)

    Fidelis, V. V.

    2011-06-01

    The observational data concerning variations of light curves of supernovae remnants—the Crab Nebula, Cassiopeia A, Tycho Brahe, and pulsar Vela—over 14 days scale that may be attributed to systematic errors of the ASM/RXTE monitor are presented. The experimental systematic errors of the GT-48 γ-ray telescope in the mono mode of operation were also determined. For this the observational data of TeV J2032 + 4130 (Cyg γ-2, according to the Crimean version) were used and the stationary nature of its γ-ray emission was confirmed by long-term observations performed with HEGRA and MAGIC. The results of research allow us to draw the following conclusions: (1) light curves of supernovae remnants averaged for long observing periods have false statistically significant flux variations, (2) the level of systematic errors is proportional to the registered flux and decreases with increasing temporal scale of averaging, (3) the light curves of sources may be modulated by the year period, and (4) the systematic errors of the GT-48 γ-ray telescope, in the amount caused by observations in the mono mode and data processing with the stereo-algorithm come to 0.12 min-1.

  13. Combination brain and systemic injections of AAV provide maximal functional and survival benefits in the Niemann-Pick mouse

    PubMed Central

    Passini, Marco A.; Bu, Jie; Fidler, Jonathan A.; Ziegler, Robin J.; Foley, Joseph W.; Dodge, James C.; Yang, Wendy W.; Clarke, Jennifer; Taksir, Tatyana V.; Griffiths, Denise A.; Zhao, Michael A.; O'Riordan, Catherine R.; Schuchman, Edward H.; Shihabuddin, Lamya S.; Cheng, Seng H.

    2007-01-01

    Niemann-Pick disease (NPD) is caused by the loss of acid sphingomyelinase (ASM) activity, which results in widespread accumulation of undegraded lipids in cells of the viscera and CNS. In this study, we tested the effect of combination brain and systemic injections of recombinant adeno-associated viral vectors encoding human ASM (hASM) in a mouse model of NPD. Animals treated by combination therapy exhibited high levels of hASM in the viscera and brain, which resulted in near-complete correction of storage throughout the body. This global reversal of pathology translated to normal weight gain and superior recovery of motor and cognitive functions compared to animals treated by either brain or systemic injection alone. Furthermore, animals in the combination group did not generate antibodies to hASM, demonstrating the first application of systemic-mediated tolerization to improve the efficacy of brain injections. All of the animals treated by combination therapy survived in good health to an investigator-selected 54 weeks, whereas the median lifespans of the systemic-alone, brain-alone, or untreated ASM knockout groups were 47, 48, and 34 weeks, respectively. These data demonstrate that combination therapy is a promising therapeutic modality for treating NPD and suggest a potential strategy for treating disease indications that cause both visceral and CNS pathologies. PMID:17517638

  14. A Retrospective Look at 20 Years of ASM Education Programs (1990–2010) and a Prospective Look at the Next 20 Years (2011–2030)

    PubMed Central

    Chang, Amy

    2011-01-01

    Professional societies provide visibility and legitimacy to the work of their post secondary educator members, advocate best practices in courses and sponsored student research, and establish deep networks and communities that catalyze members to collectively engage in undergraduate teaching and learning scholarship. Within the American Society for Microbiology (ASM), the Education Board, established in the mid-1970s, assumes this role. I have been fortunate enough to watch several pivotal programs support our growth and change the status quo by providing opportunities for biology educators to flourish. In this retrospective review, the background and details I offer about each initiative help explain ASM Education offerings, how our growth has been supported and how the status quo has changed. In this prospective look, I offer my vision of the future in post secondary education where classroom learning is student-centered and focused on global problems affecting our health and environment. For the profession to proliferate, the ASM must provide members as many opportunities in learning biology as they do with advancing biology to new frontiers. PMID:23653733

  15. Implementation of ASME Code, Section XI, Code Case N-770, on Alternative Examination Requirements for Class 1 Butt Welds Fabricated with Alloy 82/182

    SciTech Connect

    Sullivan, Edmund J.; Anderson, Michael T.

    2012-09-17

    In May 2010, the NRC issued a proposed notice of rulemaking that includes a provision to add a new section to its rules to require licensees to implement ASME Code Case N-770, ‘‘Alternative Examination Requirements and Acceptance Standards for Class 1 PWR Piping and Vessel Nozzle Butt Welds Fabricated with UNS N06082 or UNS W86182 Weld Filler Material With or Without the Application of Listed Mitigation Activities, Section XI, Division 1,’’ with 15 conditions. Code Case N-770 contains baseline and inservice inspection (ISI) requirements for unmitigated butt welds fabricated with Alloy 82/182 material and preservice and ISI requirements for mitigated butt welds. The NRC stated that application of ASME Code Case N-770 is necessary because the inspections currently required by the ASME Code, Section XI, were not written to address stress corrosion cracking Alloy 82/182 butt welds, and the safety consequences of inadequate inspections can be significant. The NRC expects to issue the final rule incorporating this code case into its regulations in the spring 2011 time frame. This paper discusses the new examination requirements, the conditions that NRC is imposing , and the major concerns with implementation of the new Code Case.

  16. Case study of the propagation of a small flaw under PWR loading conditions and comparison with the ASME code design life. Comparison of ASME Code Sections III and XI

    SciTech Connect

    Yahr, G.T.; Gwaltney, R.C.; Richardson, A.K.; Server, W.L.

    1986-01-01

    A cooperative study was performed by EG and G Idaho, Inc., and Oak Ridge National Laboratory to investigate the degree of conservatism and consistency in the ASME Boiler and Pressure Vessel Code Section III fatigue evaluation procedure and Section XI flaw acceptance standards. A single, realistic, sample problem was analyzed to determine the significance of certain points of criticism made of an earlier parametric study by staff members of the Division of Engineering Standards of the Nuclear Regulatory Commission. The problem was based on a semielliptical flaw located on the inside surface of the hot-leg piping at the reactor vessel safe-end weld for the Zion 1 pressurized-water reactor (PWR). Two main criteria were used in selecting the problem; first, it should be a straight pipe to minimize the computational expense; second, it should exhibit as high a cumulative usage factor as possible. Although the problem selected has one of the highest cumulative usage factors of any straight pipe in the primary system of PWRs, it is still very low. The Code Section III fatigue usage factor was only 0.00046, assuming it was in the as-welded condition, and fatigue crack-growth analyses predicted negligible crack growth during the 40-year design life. When the analyses were extended past the design life, the usage factor was less than 1.0 when the flaw had propagated to failure. The current study shows that the criticism of the earlier report should not detract from the conclusion that if a component experiences a high level of cyclic stress corresponding to a fatigue usage factor near 1.0, very small cracks can propagate to unacceptable sizes.

  17. Results from Evaluation of Representative ASME AG-1 Section FK Radial Flow Dimple Pleated HEPA Filters Under Elevated Conditions - 12002

    SciTech Connect

    Giffin, Paxton K.; Parsons, Michael S.; Rickert, Jaime G.; Waggoner, Charles A.

    2012-07-01

    The American Society of Mechanical Engineers (ASME) has recently added Section FK establishing requirements for radial flow HEPA filters to the Code on Nuclear Air and Gas Treatment (AG-1). Section FK filters are expected to be a major element in the HEPA filtration systems across the US Department of Energy (DOE) complex. Radial flow filters have been used in Europe for some time, however a limited amount of performance evaluation data exists with respect to these new AG-1 Section FK units. In consultation with a technical working group, the Institute for Clean Energy Technology (ICET) at Mississippi State University (MSU)has evaluated a series of representative AG-1 Section FK dimple pleated radial flow HEPA filters. The effects of elevated relative humidity and temperature conditions on these filters are particularly concerning. Results from the evaluation of Section FK filters under ambient conditions have been presented at the 2011 waste management conference. Additions to the previous test stand to enable high temperature and high humidity testing, a review of the equipment used, the steps taken to characterize the new additions, and the filter test results are presented in this study. Test filters were evaluated at a volumetric flow rate of 56.6 m{sup 3}/min (2000 cfm) and were challenged under ambient conditions with Alumina, Al(OH){sub 3}, until reaching a differential pressure of 1 kPa (4 in. w.c.), at which time the filters were tested, unchallenged with aerosol, at 54 deg. C (130 deg. F) for approximately 1 hour. At the end of that hour water was sprayed near the heat source to maximize vaporization exposing the filter to an elevated relative humidity up to 95%. Collected data include differential pressure, temperature, relative humidity, and volumetric flow rate versus time. (authors)

  18. Deregulation of sphingolipid metabolism in Alzheimer's disease

    PubMed Central

    He, Xingxuan; Huang, Yu; Li, Bin; Gong, Cheng-Xing; Schuchman, Edward H.

    2010-01-01

    Abnormal sphingolipid metabolism has been previously reported in Alzheimer's disease (AD). To extend these findings, several sphingolipids and sphingolipid hydrolases were analyzed in brain samples from AD patients and age-matched normal individuals. We found a pattern of elevated acid sphingomyelinase (ASM) and acid ceramidase (AC) expression in AD, leading to a reduction in sphingomyelin and elevation of ceramide. More sphingosine also was found in the AD brains, although sphingosine-1-phosphate (S1P) levels were reduced. Notably, significant correlations were observed between the brain ASM and S1P levels and the levels of amyloid beta peptide (Aβ) and phosphorylated tau protein. Based on these findings, neuronal cell cultures were treated with Aβ oligomers, which were found to activate ASM, increase ceramide, and induce apoptosis. Pre-treatment of the neurons with purified, recombinant AC prevented the cells from undergoing Aβ-induced apoptosis. We propose that ASM activation is an important pathological event leading to AD, perhaps due to Aβ deposition. The downstream consequences of ASM activation are elevated ceramide, activation of ceramidases, and production of sphingosine. The reduced levels of S1P in the AD brain, together with elevated ceramide, likely contribute to the disease pathogenesis. PMID:18547682

  19. A study of outburst ephemeris and burst properties of blackhole candidate 4U 1630-47 with ASM, MAXI and Suzaku data

    NASA Astrophysics Data System (ADS)

    Abraham, Lalitha; Agrawal, V. K.

    4U 1630-47 is a soft X-ray transient which is thought to be a blackhole candidate. This source exhibits quasi-periodic outbursts on time scales of 500-700 days. In addition to the normal outbursts which usually last for a few months, the source displays superoutbursts, lasting for one to two years, seen to recur in every 10-12 years. The outburst ephemeris has been studied previously upto 1996 outbursts. In this work we present the updated ephemeris using 16 years data obtained from All Sky Monitor (ASM) onboard RXTE and one years data from MAXI satellite. The data covers 7 outbursts seen from ASM and one outburst seen by MAXI. We study morphology of each of these outbursts. We find that most of the bursts can be classified in basic three categories: flat top, FRED (Fast Rise Exponential Decay) and triangular. We also investigate relation between burst properties with quiescent flux level using Suzaku data, a study which has not been done previously.

  20. ITER's Tokamak Cooling Water System and the the Use of ASME Codes to Comply with French Regulations of Nuclear Pressure Equipment

    SciTech Connect

    Berry, Jan; Ferrada, Juan J; Curd, Warren; Dell Orco, Dr. Giovanni; Barabash, Vladimir; Kim, Seokho H

    2011-01-01

    During inductive plasma operation of ITER, fusion power will reach 500 MW with an energy multiplication factor of 10. The heat will be transferred by the Tokamak Cooling Water System (TCWS) to the environment using the secondary cooling system. Plasma operations are inherently safe even under the most severe postulated accident condition a large, in-vessel break that results in a loss-of-coolant accident. A functioning cooling water system is not required to ensure safe shutdown. Even though ITER is inherently safe, TCWS equipment (e.g., heat exchangers, piping, pressurizers) are classified as safety important components. This is because the water is predicted to contain low-levels of radionuclides (e.g., activated corrosion products, tritium) with activity levels high enough to require the design of components to be in accordance with French regulations for nuclear pressure equipment, i.e., the French Order dated 12 December 2005 (ESPN). ESPN has extended the practical application of the methodology established by the Pressure Equipment Directive (97/23/EC) to nuclear pressure equipment, under French Decree 99-1046 dated 13 December 1999, and Order dated 21 December 1999 (ESP). ASME codes and supplementary analyses (e.g., Failure Modes and Effects Analysis) will be used to demonstrate that the TCWS equipment meets these essential safety requirements. TCWS is being designed to provide not only cooling, with a capacity of approximately 1 GW energy removal, but also elevated temperature baking of first-wall/blanket, vacuum vessel, and divertor. Additional TCWS functions include chemical control of water, draining and drying for maintenance, and facilitation of leak detection/localization. The TCWS interfaces with the majority of ITER systems, including the secondary cooling system. U.S. ITER is responsible for design, engineering, and procurement of the TCWS with industry support from an Engineering Services Organization (ESO) (AREVA Federal Services, with support

  1. Optimized periodic verification testing blended risk and performance-based MOV inservice test program an application of ASME code case OMN-1

    SciTech Connect

    Sellers, C.; Fleming, K.; Bidwell, D.; Forbes, P.

    1996-12-01

    This paper presents an application of ASME Code Case OMN-1 to the GL 89-10 Program at the South Texas Project Electric Generating Station (STPEGS). Code Case OMN-1 provides guidance for a performance-based MOV inservice test program that can be used for periodic verification testing and allows consideration of risk insights. Blended probabilistic and deterministic evaluation techniques were used to establish inservice test strategies including both test methods and test frequency. Described in the paper are the methods and criteria for establishing MOV safety significance based on the STPEGS probabilistic safety assessment, deterministic considerations of MOV performance characteristics and performance margins, the expert panel evaluation process, and the development of inservice test strategies. Test strategies include a mix of dynamic and static testing as well as MOV exercising.

  2. Review of ASME code criteria for control of primary loads on nuclear piping system branch connections and recommendations for additional development work

    SciTech Connect

    Rodabaugh, E.C.; Gwaltney, R.C.; Moore, S.E.

    1993-11-01

    This report collects and uses available data to reexamine the criteria for controlling primary loads in nuclear piping branch connections as expressed in Section III of the ASME Boiler and Pressure Vessel Code. In particular, the primary load stress indices given in NB-3650 and NB-3683 are reexamined. The report concludes that the present usage of the stress indices in the criteria equations should be continued. However, the complex treatment of combined branch and run moments is not supported by available information. Therefore, it is recommended that this combined loading evaluation procedure be replaced for primary loads by the separate leg evaluation procedure specified in NC/ND-3653.3(c) and NC/ND-3653.3(d). No recommendation is made for fatigue or secondary load evaluations for Class 1 piping. Further work should be done on the development of better criteria for treatment of combined branch and run moment effects.

  3. First-principles study of energetic and electronic properties of A2Ti2O7 (A=Sm, Gd, Er) pyrochlore

    SciTech Connect

    Xiao, H. Y.; Zu, Xiaotao T.; Gao, Fei; Weber, William J.

    2008-10-01

    First-principles calculations have been carried out to study the electronic properties of A2Ti2O7 (A=Sm, Gd, Er) pyrochlores. It was found that f electrons have negligible effect on the structural and energetic properties, but have significant effect on the electronic properties. Density of state analysis shows that A-site 4f electrons do take part in the chemical bonding. Also, we found that bond is less covalent than and bonds, while bond in Gd2Ti2O7 is more covalent. It was proposed that for A2Ti2O7 (A = Sm, Gd, Er) pyrochlores, bonds may play more significant role in determining their radiation resistance to amorphization.

  4. Probing the Mysteries of the X-Ray Binary 4U 1210-64 with ASM, PCA, MAXI, BAT, and Suzaku

    NASA Astrophysics Data System (ADS)

    Coley, Joel B.; Corbet, Robin H. D.; Mukai, Koji; Pottschmidt, Katja

    2014-10-01

    4U 1210-64 has been postulated to be a high-mass X-ray binary powered by the Be mechanism. X-ray observations with Suzaku, the ISS Monitor of All-sky X-ray Image (MAXI), and the Rossi X-ray Timing Explorer Proportional Counter Array (PCA) and All Sky Monitor (ASM) provide detailed temporal and spectral information on this poorly understood source. Long-term ASM and MAXI observations show distinct high and low states and the presence of a 6.7101 ± 0.0005 day modulation, interpreted as the orbital period. Folded light curves reveal a sharp dip, interpreted as an eclipse. To determine the nature of the mass donor, the predicted eclipse half-angle was calculated as a function of inclination angle for several stellar spectral types. The eclipse half-angle is not consistent with a mass donor of spectral type B5 V however, stars with spectral types B0 V or B0-5 III are possible. The best-fit spectral model consists of a power law with index Γ = 1.85^{+0.04}_{-0.05} and a high-energy cutoff at 5.5 ± 0.2 keV modified by an absorber that fully covers the source as well as partially covering absorption. Emission lines from S XVI Kα, Fe Kα, Fe XXV Kα, and Fe XXVI Kα were observed in the Suzaku spectra. Out of eclipse, the Fe Kα line flux was strongly correlated with unabsorbed continuum flux, indicating that the Fe I emission is the result of fluorescence of cold dense material near the compact object. The Fe I feature is not detected during eclipse, further supporting an origin close to the compact object.

  5. Probing the mysteries of the X-ray binary 4U 1210-64 with ASM, PCA, MAXI, BAT, and Suzaku

    SciTech Connect

    Coley, Joel B.; Corbet, Robin H. D.; Mukai, Koji; Pottschmidt, Katja

    2014-10-01

    4U 1210-64 has been postulated to be a high-mass X-ray binary powered by the Be mechanism. X-ray observations with Suzaku, the ISS Monitor of All-sky X-ray Image (MAXI), and the Rossi X-ray Timing Explorer Proportional Counter Array (PCA) and All Sky Monitor (ASM) provide detailed temporal and spectral information on this poorly understood source. Long-term ASM and MAXI observations show distinct high and low states and the presence of a 6.7101 ± 0.0005 day modulation, interpreted as the orbital period. Folded light curves reveal a sharp dip, interpreted as an eclipse. To determine the nature of the mass donor, the predicted eclipse half-angle was calculated as a function of inclination angle for several stellar spectral types. The eclipse half-angle is not consistent with a mass donor of spectral type B5 V; however, stars with spectral types B0 V or B0-5 III are possible. The best-fit spectral model consists of a power law with index Γ = 1.85{sub −0.05}{sup +0.04} and a high-energy cutoff at 5.5 ± 0.2 keV modified by an absorber that fully covers the source as well as partially covering absorption. Emission lines from S XVI Kα, Fe Kα, Fe XXV Kα, and Fe XXVI Kα were observed in the Suzaku spectra. Out of eclipse, the Fe Kα line flux was strongly correlated with unabsorbed continuum flux, indicating that the Fe I emission is the result of fluorescence of cold dense material near the compact object. The Fe I feature is not detected during eclipse, further supporting an origin close to the compact object.

  6. RXTE/ASM and Swift/BAT observations of spectral transitions in bright X-ray binaries in 2005-2010

    NASA Astrophysics Data System (ADS)

    Tang, Jing; Yu, Wen-Fei; Yan, Zhen

    2011-04-01

    We have studied X-ray spectral state transitions that can be seen in the long-term monitoring light curves of bright X-ray binaries from the All-Sky Monitor (ASM) onboard the Rossi X-ray Timing Explorer (RXTE) and the Burst Alert Telescope (BAT) onboard Swift during a period of five years from 2005 to 2010. We have applied a program to automatically identify the hard-to-soft (H-S) spectral state transitions in the bright X-ray binaries monitored by the ASM and the BAT. In total, we identified 128 hard-to-soft transitions, of which 59 occurred after 2008. We also determined the transition fluxes and the peak fluxes of the following soft states, updated the measurements of the luminosity corresponding to the H-S transition and the peak luminosity of the following soft state in about 30 bright persistent and transient black hole and neutron star binaries following Yu & Yan, and found the luminosity correlation and the luminosity range of spectral transitions in data between 2008-2010 are about the same as those derived from data before 2008. This further strengthens the idea that the luminosity at which the H-S spectral transition occurs in the Galactic X-ray binaries is determined by non-stationary accretion parameters such as the rate-of-change of the mass accretion rate rather than the mass accretion rate itself. The correlation is also found to hold in data of individual sources 4U 1608-52 and 4U 1636-53.

  7. Ratio of Dietary n-6/n-3 Polyunsaturated Fatty Acids Independently Related to Muscle Mass Decline in Hemodialysis Patients

    PubMed Central

    Wong, Te-Chih; Chen, Yu-Tong; Wu, Pei-Yu; Chen, Tzen-Wen; Chen, Hsi-Hsien; Chen, Tso-Hsiao; Yang, Shwu-Huey

    2015-01-01

    Background n-3 polyunsaturated fatty acids (PUFAs) might be useful nutritional strategy for treating patients with sarcopenia. We evaluated the effect of the intake of dietary n-3 PUFAs on the skeletal muscle mass (SMM), appendicular skeletal muscle mass (ASM), and its determinants in patients receiving standard hemodialysis (HD) treatment for the management of end stage renal disease. Methods In this cross-sectional study, data of 111 HD patients were analyzed. Anthropometric and bioelectrical impedance measurements used to estimate the muscle mass were performed the day of dialysis immediately after the dialysis session. Routine laboratory and 3-day dietary data were also collected. The cutoff value of adequate intake (AI) for both n-3 PUFAs and alpha-linolenic acid (ALA) was 1.6 g/day and 1.1 g/day for men and women, respectively. Results The mean age, mean dietary n-3 PUFAs intake, ALA intake, ratio of n-6/n-3 PUFAs intake, SMM, and ASM of patients were 61.4 ± 10.4 years, 2.0 ± 1.3 g/day, 1.5 ± 1.0 g/day, 9.5 ± 6.7 g/day, 23.9 ± 5.5 kg, and 17.5 ± 4.5 kg, respectively. A higher SMM and ASM significantly observed in patients who achieved an AI of n-3 PUFAs. Similar trends appeared to be observed among those patients who achieved the AI of ALA, but the difference was not significantly, except for ASM (P = 0.047). No relevant differences in demographics, laboratory and nutritional parameters were observed, regardless of whether the patients achieved an AI of n-3 PUFAs. Multivariate analysis showed that the BMI and equilibrated Kt/V were independent determinants of the muscle mass. Moreover, the ratio of n-6/n-3 PUFAs was an independent risk determinant of reduced ASM in HD patients. Conclusion Patients with an AI of n-3 PUFAs had better total-body SMM and ASM. A higher dietary ratio of n-6/n-3 PUFAs seemed to be associated with a reduced muscle mass in HD patients. PMID:26466314

  8. Genetics of Lipid-Storage Management in Caenorhabditis elegans Embryos.

    PubMed

    Schmökel, Verena; Memar, Nadin; Wiekenberg, Anne; Trotzmüller, Martin; Schnabel, Ralf; Döring, Frank

    2016-03-01

    Lipids play a pivotal role in embryogenesis as structural components of cellular membranes, as a source of energy, and as signaling molecules. On the basis of a collection of temperature-sensitive embryonic lethal mutants, a systematic database search, and a subsequent microscopic analysis of >300 interference RNA (RNAi)-treated/mutant worms, we identified a couple of evolutionary conserved genes associated with lipid storage in Caenorhabditis elegans embryos. The genes include cpl-1 (cathepsin L-like cysteine protease), ccz-1 (guanine nucleotide exchange factor subunit), and asm-3 (acid sphingomyelinase), which is closely related to the human Niemann-Pick disease-causing gene SMPD1. The respective mutant embryos accumulate enlarged droplets of neutral lipids (cpl-1) and yolk-containing lipid droplets (ccz-1) or have larger genuine lipid droplets (asm-3). The asm-3 mutant embryos additionally showed an enhanced resistance against C band ultraviolet (UV-C) light. Herein we propose that cpl-1, ccz-1, and asm-3 are genes required for the processing of lipid-containing droplets in C. elegans embryos. Owing to the high levels of conservation, the identified genes are also useful in studies of embryonic lipid storage in other organisms. PMID:26773047

  9. Updating of ASME Nuclear Code Case N-201 to Accommodate the Needs of Metallic Core Support Structures for High Temperature Gas Cooled Reactors Currently in Development

    SciTech Connect

    Mit Basol; John F. Kielb; John F. MuHooly; Kobus Smit

    2007-05-02

    On September 29, 2005, ASME Standards Technology, LLC (ASME ST-LLC) executed a multi-year, cooperative agreement with the United States DOE for the Generation IV Reactor Materials project. The project's objective is to update and expand appropriate materials, construction, and design codes for application in future Generation IV nuclear reactor systems that operate at elevated temperatures. Task 4 was embarked upon in recognition of the large quantity of ongoing reactor designs utilizing high temperature technology. Since Code Case N-201 had not seen a significant revision (except for a minor revision in September, 2006 to change the SA-336 forging reference for 304SS and 316SS to SA-965 in Tables 1.2(a) and 1.2(b), and some minor editorial changes) since December 1994, identifying recommended updates to support the current high temperature Core Support Structure (CSS) designs and potential new designs was important. As anticipated, the Task 4 effort identified a number of Code Case N-201 issues. Items requiring further consideration range from addressing apparent inconsistencies in definitions and certain material properties between CC-N-201 and Subsection NH, to inclusion of additional materials to provide the designer more flexibility of design. Task 4 developed a design parameter survey that requested input from the CSS designers of ongoing high temperature gas cooled reactor metallic core support designs. The responses to the survey provided Task 4 valuable input to identify the design operating parameters and future needs of the CSS designers. Types of materials, metal temperature, time of exposure, design pressure, design life, and fluence levels were included in the Task 4 survey responses. The results of the survey are included in this report. This research proves that additional work must be done to update Code Case N-201. Task 4 activities provide the framework for the Code Case N-201 update and future work to provide input on materials. Candidate

  10. Biological activity of palladium(II) and platinum(II) complexes of the acetone Schiff bases of S-methyl- and S-benzyldithiocarbazate and the X-ray crystal structure of the [Pd(asme)2] (asme=anionic form of the acetone Schiff base of S-methyldithiocarbazate) complex.

    PubMed

    Akbar Ali, Mohammad; Mirza, Aminul Huq; Butcher, Raymond J; Tarafder, M T H; Keat, Tan Boon; Ali, A Manaf

    2002-11-25

    Palladium(II) and platinum(II) complexes of general empirical formula, [M(NS)(2)] (NS=uninegatively charged acetone Schiff bases of S-methyl- and S-benzyldithiocarbazate; M=Pt(II) and Pd(II)) have been prepared and characterized by a variety of physicochemical techniques. Based on conductance, IR and electronic spectral evidence, a square-planar structure is assigned to these complexes. The crystal and molecular structure of the [Pd(asme)(2)] complex (asme=anionic form of the acetone Schiff base of S-methyldithiocarbazate) has been determined by X-ray diffraction. The complex has a distorted cis-square planar structure with the ligands coordinated to the palladium(II) ions as uninegatively charged bidentate NS chelating agents via the azomethine nitrogen and the mercaptide sulfur atoms. The distortion from a regular square-planar geometry is attributed to the restricted bite angles of the ligands. Antimicrobial tests indicate that the Schiff bases exhibit strong activities against the pathogenic bacteria, Bacillus subtilis (mutant defective DNA repair), methicillin-resistant Staphylococcus aureus, B. subtilis (wild type) and Pseudomonas aeruginosa and the fungi, Candida albicans (CA), Candida lypotica (2075), Saccharomyces cerevisiae (20341) and Aspergillus ochraceous (398)-the activities exhibited by these compounds being greater than that of the standard antibacterial and antifungal drugs, streptomycin and nystatin, respectively. The palladium(II) and platinum(II) complexes are inactive against most of these organisms but, the microbe, Pseudomonas aeruginosa shows strong sensitivity to the platinum(II) complexes. Screening of the compounds for their cytotoxicities against T-lymphoblastic leukemia cancer cells has shown that the acetone Schiff base of S-methyldithiocarbazate (Hasme) exhibits a very weak activity, whereas the S-benzyl derivative (Hasbz) is inactive. However, the palladium(II) complexes exhibit strong cytotoxicities against this cancer; their

  11. Parameter estimation procedure for complex non-linear systems: calibration of ASM No. 1 for N-removal in a full-scale oxidation ditch.

    PubMed

    Abusam, A; Keesman, K J; van Straten, G; Spanjers, H; Meinema, K

    2001-01-01

    When applied to large simulation models, the process of parameter estimation is also called calibration. Calibration of complex non-linear systems, such as activated sludge plants, is often not an easy task. On the one hand, manual calibration of such complex systems is usually time-consuming, and its results are often not reproducible. On the other hand, conventional automatic calibration methods are not always straightforward and often hampered by local minima problems. In this paper a new straightforward and automatic procedure, which is based on the response surface method (RSM) for selecting the best identifiable parameters, is proposed. In RSM, the process response (output) is related to the levels of the input variables in terms of a first- or second-order regression model. Usually, RSM is used to relate measured process output quantities to process conditions. However, in this paper RSM is used for selecting the dominant parameters, by evaluating parameters sensitivity in a predefined region. Good results obtained in calibration of ASM No. 1 for N-removal in a full-scale oxidation ditch proved that the proposed procedure is successful and reliable. PMID:11385868

  12. ASME Section VIII Recertification of a 33,000 Gallon Vacuum-jacketed LH2 Storage Vessel for Densified Hydrogen Testing at NASA Kennedy Space Center

    NASA Technical Reports Server (NTRS)

    Swanger, Adam M.; Notardonato, William U.; Jumper, Kevin M.

    2015-01-01

    The Ground Operations Demonstration Unit for Liquid Hydrogen (GODU-LH2) has been developed at NASA Kennedy Space Center in Florida. GODU-LH2 has three main objectives: zero-loss storage and transfer, liquefaction, and densification of liquid hydrogen. A cryogenic refrigerator has been integrated into an existing, previously certified, 33,000 gallon vacuum-jacketed storage vessel built by Minnesota Valley Engineering in 1991 for the Titan program. The dewar has an inner diameter of 9.5 and a length of 71.5; original design temperature and pressure ranges are -423 F to 100 F and 0 to 95 psig respectively. During densification operations the liquid temperature will be decreased below the normal boiling point by the refrigerator, and consequently the pressure inside the inner vessel will be sub-atmospheric. These new operational conditions rendered the original certification invalid, so an effort was undertaken to recertify the tank to the new pressure and temperature requirements (-12.7 to 95 psig and -433 F to 100 F respectively) per ASME Boiler and Pressure Vessel Code, Section VIII, Division 1. This paper will discuss the unique design, analysis and implementation issues encountered during the vessel recertification process.

  13. Periodic X-ray Modulation and its Possible Relation with Eccentricity in Black Hole Binaries : Long-Term Swift/BAT and RXTE/ASM Data Analysis

    NASA Astrophysics Data System (ADS)

    Ghosh, Arindam; Chakrabarti, Sandip Kumar

    2016-07-01

    X-ray binary orbits are expected to have some eccentricity, albeit small. Stellar companion of a black hole orbiting in an eccentric orbit will experience modulating tidal force with a periodicity same as that of the orbital period which will result in a modulation of accretion rates, seed photon flux, and flux of inverse Comptonized harder X-rays as well. Timing analysis of long-term X-ray data (1.5-12 keV) of RXTE/ASM and all sky survey data (15-50 keV) of Swift/BAT satellites reveal this periodicity in several black hole candidates. If this modulation is assumed to be solely due to tidal effects (without taking other effects, such as eclipses, reflection from winds, super-hump phenomena etc. into account), the RMS-value of the peak in power density spectrum allows us to estimate eccentricities of these orbits. We present these very interesting results. We show that our results generally agree with independent studies of these parameters.

  14. Evidence of Two Component Accretion Flows as revealed by time lag properties: Results of Long-Term RXTE/ASM Data Analysis

    NASA Astrophysics Data System (ADS)

    Ghosh, Arindam; Chakrabarti, Sandip Kumar

    2016-07-01

    Long-term RXTE/ASM X-ray data of several Galactic black hole candidates (BHCs) are analyzed. The results of this analysis show the existence of two component accretion flow (TCAF) in both low-mass and high-mass X-ray binaries (LMXBs & HMXBs). Large disks with long viscous timescales in the accreting matter with high angular momentum are prevalent in LMXBs due to processes like Roche lobe overflow, while small disks with little viscous delays are observed in HMXBs, primarily because of wind accretion. Two parameters are defined as photon indices, independent of the choice of a BHC, in order to find correlation between the two components, namely, the Keplerian disk component and the sub-Keplerian component, thereby estimating the time lag between two aforesaid timescales. Fluxes of hard and soft photons are observed to be anti-correlated with respect to these photon indices. The time lags give us an idea of the viscosity in the Keplerian component.

  15. A Comparison of the Variability of the Symbiotic X-ray Binaries GX 1+4, 4U 1954+31, and 4U 1700+24 from Swift/BAT and RXTE/ASM Observations

    NASA Technical Reports Server (NTRS)

    Corbet, R. H. D.; Sokoloski, J. L.; Mukai, K.; Markwardt, C. B.; Tueller, J.

    2007-01-01

    We present an analysis of the X-ray variability of three symbiotic X-ray binaries, GX 1+4, 4U 1700+24, and 4U 1954+31, using observations made with the Swift Burst Alert Telescope (BAT) and the Rossi X-ray Timing Explorer (RXTE) All-Sky Monitor (ASM). Observations of 4U 1954+31 with the Swift BAT show modulation at a period near 5 hours. Models to explain this modulation are discussed including the presence of an exceptionally slow X-ray pulsar in the system and accretion instabilities. We conclude that the most likely interpretation is that 4U 1954+31 contains one of the slowest known X-ray pulsars. Unlike 4U 1954+31, neither GX 1+4 nor 4U 1700+24 show any evidence for modulation on a timescale of hours. An analysis of the RXTE ASM light curves of GX l+4, 4U 1700+24, and 4U 1954+31 does not show the presence of periodic modulation in any source, although there is considerable variability on long timescales for all three sources. There is no modulation in GX 1+4 on either the optical 1161 day orbital period or a previously reported 304 day X-ray period. For 4U 1700+24 we do not confirm the 404 day period previously proposed for this source from a shorter duration ASM light curve.

  16. A sphingolipid mechanism for behavioral extinction.

    PubMed

    Huston, Joseph P; Kornhuber, Johannes; Mühle, Christiane; Japtok, Lukasz; Komorowski, Mara; Mattern, Claudia; Reichel, Martin; Gulbins, Erich; Kleuser, Burkhard; Topic, Bianca; De Souza Silva, Maria A; Müller, Christian P

    2016-05-01

    Reward-dependent instrumental behavior must continuously be re-adjusted according to environmental conditions. Failure to adapt to changes in reward contingencies may incur psychiatric disorders like anxiety and depression. When an expected reward is omitted, behavior undergoes extinction. While extinction involves active re-learning, it is also accompanied by emotional behaviors indicative of frustration, anxiety, and despair (extinction-induced depression). Here, we report evidence for a sphingolipid mechanism in the extinction of behavior. Rapid extinction, indicating efficient re-learning, coincided with a decrease in the activity of the enzyme acid sphingomyelinase (ASM), which catalyzes turnover of sphingomyelin to ceramide, in the dorsal hippocampus of rats. The stronger the decline in ASM activity, the more rapid was the extinction. Sphingolipid-focused lipidomic analysis showed that this results in a decline of local ceramide species in the dorsal hippocampus. Ceramides shape the fluidity of lipid rafts in synaptic membranes and by that way can control neural plasticity. We also found that aging modifies activity of enzymes and ceramide levels in selective brain regions. Aging also changed how the chronic treatment with corticosterone (stress) or intranasal dopamine modified regional enzyme activity and ceramide levels, coinciding with rate of extinction. These data provide first evidence for a functional ASM-ceramide pathway in the brain involved in the extinction of learned behavior. This finding extends the known cellular mechanisms underlying behavioral plasticity to a new class of membrane-located molecules, the sphingolipids, and their regulatory enzymes, and may offer new treatment targets for extinction- and learning-related psychopathological conditions. Sphingolipids are common lipids in the brain which form lipid domains at pre- and postsynaptic membrane compartments. Here we show a decline in dorsal hippocampus ceramide species together with a

  17. Amino acids

    MedlinePlus

    Amino acids are organic compounds that combine to form proteins . Amino acids and proteins are the building blocks of life. When proteins are digested or broken down, amino acids are left. The human body uses amino acids ...

  18. Acute Acidification of Stratum Corneum Membrane Domains Using Polyhydroxyl Acids Improves Lipid Processing and Inhibits Degradation of Corneodesmosomes

    PubMed Central

    Hachem, Jean-Pierre; Roelandt, Truus; Schürer, Nanna; Pu, Xu; Fluhr, Joachim; Giddelo, Christina; Man, Mao-Qiang; Crumrine, Debra; Roseeuw, Diane; Feingold, Kenneth R.; Mauro, Theodora; Elias, Peter M.

    2010-01-01

    Neutralization of the normally acidic stratum corneum (SC) has deleterious consequences for permeability barrier homeostasis and SC integrity/cohesion attributable to serine proteases (SPs) activation leading to deactivation/degradation of lipid-processing enzymes and corneodesmosomes (CD). As an elevated pH compromises SC structure and function, we asked here whether SC hyperacidification would improve the structure and function. We lowered the pH of mouse SC using two polyhydroxyl acids (PHA), lactobionic acid (LBA), or gluconolactone (GL). Applications of the PHA reduced the pH at all levels of SC of hairless mouse, with further selective acidification of SC membrane domains, as shown by fluorescence lifetime imaging. Hyperacidification improved permeability barrier homeostasis, attributable to increased activities of two key membrane-localized, ceramide-generating hydrolytic enzymes (β-glucocerebrosidase and acidic sphingomyelinase), which correlated with accelerated extracellular maturation of SC lamellar membranes. Hyperacidification generated “supernormal” SC integrity/cohesion, attributable to an SP-dependent decreased degradation of desmoglein-1 (DSG1) and the induction of DSG3 expression in lower SC. As SC hyperacidification improves the structure and function, even of normal epidermis, these studies lay the groundwork for an assessment of the potential utility of SC acidification as a therapeutic strategy for inflammatory dermatoses, characterized by abnormalities in barrier function, cohesion, and surface pH. PMID:19741713

  19. Acute acidification of stratum corneum membrane domains using polyhydroxyl acids improves lipid processing and inhibits degradation of corneodesmosomes.

    PubMed

    Hachem, Jean-Pierre; Roelandt, Truus; Schürer, Nanna; Pu, Xu; Fluhr, Joachim; Giddelo, Christina; Man, Mao-Qiang; Crumrine, Debra; Roseeuw, Diane; Feingold, Kenneth R; Mauro, Theodora; Elias, Peter M

    2010-02-01

    Neutralization of the normally acidic stratum corneum (SC) has deleterious consequences for permeability barrier homeostasis and SC integrity/cohesion attributable to serine proteases (SPs) activation leading to deactivation/degradation of lipid-processing enzymes and corneodesmosomes (CD). As an elevated pH compromises SC structure and function, we asked here whether SC hyperacidification would improve the structure and function. We lowered the pH of mouse SC using two polyhydroxyl acids (PHA), lactobionic acid (LBA), or gluconolactone (GL). Applications of the PHA reduced the pH at all levels of SC of hairless mouse, with further selective acidification of SC membrane domains, as shown by fluorescence lifetime imaging. Hyperacidification improved permeability barrier homeostasis, attributable to increased activities of two key membrane-localized, ceramide-generating hydrolytic enzymes (beta-glucocerebrosidase and acidic sphingomyelinase), which correlated with accelerated extracellular maturation of SC lamellar membranes. Hyperacidification generated "supernormal" SC integrity/cohesion, attributable to an SP-dependent decreased degradation of desmoglein-1 (DSG1) and the induction of DSG3 expression in lower SC. As SC hyperacidification improves the structure and function, even of normal epidermis, these studies lay the groundwork for an assessment of the potential utility of SC acidification as a therapeutic strategy for inflammatory dermatoses, characterized by abnormalities in barrier function, cohesion, and surface pH. PMID:19741713

  20. The STEM-II regional scale acid deposition and photochemical oxidant model—I. An overview of model development and applications

    NASA Astrophysics Data System (ADS)

    Carmichael, Gregory R.; Peters, Leonard K.; Saylor, Rick D.

    The STEM-II pollutant transport/transformation/removal model has been enhanced to include scavenging by clouds and precipitation. The Advanced Scavenging Model (ASM) and Reactive Scavenging Model (RSM) have been coupled to STEM-II to enable comprehensive simulation and diagnosis analysis of field experiments designed to study acid deposition. In this paper, the STEM-II/ASM/RSM model is described. The coupled model is being used to analyze the PRECP-I field studies on an urban/suburban scale (Philadelphia), mesoscale (lower Ohio River Valley), and regional scale (eastern U.S.). The time period selected for detailed simulations and analysis is 29 April-5 May 1985. Subsequent papers will detail the results of these simulations.

  1. Folic Acid

    MedlinePlus

    Folic acid is a B vitamin. It helps the body make healthy new cells. Everyone needs folic acid. For women who may get pregnant, it is really important. Getting enough folic acid before and during pregnancy can prevent major birth ...

  2. Folic Acid

    MedlinePlus

    Folic acid is used to treat or prevent folic acid deficiency. It is a B-complex vitamin needed by ... Folic acid comes in tablets. It usually is taken once a day. Follow the directions on your prescription label ...

  3. Aspartic acid

    MedlinePlus

    ... also called asparaginic acid. Aspartic acid helps every cell in the body work. It plays a role in: Hormone production and release Normal nervous system function Plant sources of aspartic acid include: Legumes such as ...

  4. Acid Rain.

    ERIC Educational Resources Information Center

    Openshaw, Peter

    1987-01-01

    Provides some background information on acid deposition. Includes a historical perspective, describes some effects of acid precipitation, and discusses acid rain in the United Kingdom. Contains several experiments that deal with the effects of acid rain on water quality and soil. (TW)

  5. Sphingomyelin Phosphodiesterase Acid-like 3A (SMPDL3A) Is a Novel Nucleotide Phosphodiesterase Regulated by Cholesterol in Human Macrophages*

    PubMed Central

    Traini, Mathew; Quinn, Carmel M.; Sandoval, Cecilia; Johansson, Erik; Schroder, Kate; Kockx, Maaike; Meikle, Peter J.; Jessup, Wendy; Kritharides, Leonard

    2014-01-01

    Cholesterol-loaded foam cell macrophages are prominent in atherosclerotic lesions and play complex roles in both inflammatory signaling and lipid metabolism, which are underpinned by large scale reprogramming of gene expression. We performed a microarray study of primary human macrophages that showed that transcription of the sphingomyelin phosphodiesterase acid-like 3A (SMPDL3A) gene is up-regulated after cholesterol loading. SMPDL3A protein expression in and secretion from primary macrophages are stimulated by cholesterol loading, liver X receptor ligands, and cyclic AMP, and N-glycosylated SMPDL3A protein is detectable in circulating blood. We demonstrate for the first time that SMPDL3A is a functional phosphodiesterase with an acidic pH optimum. We provide evidence that SMPDL3A is not an acid sphingomyelinase but unexpectedly is active against nucleotide diphosphate and triphosphate substrates at acidic and neutral pH. SMPDL3A is a major source of nucleotide phosphodiesterase activity secreted by liver X receptor-stimulated human macrophages. Extracellular nucleotides such as ATP may activate pro-inflammatory responses in immune cells. Increased expression and secretion of SMPDL3A by cholesterol-loaded macrophage foam cells in lesions may decrease local concentrations of pro-inflammatory nucleotides and potentially represent a novel anti-inflammatory axis linking lipid metabolism with purinergic signaling in atherosclerosis. PMID:25288789

  6. Types A and B Niemann-Pick Disease.

    PubMed

    Schuchman, Edward H; Wasserstein, Melissa P

    2016-06-01

    Two distinct metabolic abnormalities are included under the eponym Niemann-Pick disease (NPD). The first is due to the deficient activity of the enzyme acid sphingomyelinase (ASM). Patients with ASM deficiency are classified as having types A and B Niemann-Pick disease (NPD). Type A NPD patients exhibit hepatosplenomegaly, frequent pulmonary infections, and profound central nervous system involvement in infancy. They rarely survive beyond two years of age. Type B patients also have hepatosplenomegaly and progressive alterations of their lungs, but there are usually no central nervous system signs. The age of onset and rate of disease progression varies greatly among type B patients, and they frequently live into adulthood. Recently, patients with phenotypes intermediate between types A and B NPD also have been identified. These individuals represent the expected continuum caused by inheriting different mutations in the ASM gene (SMPD1). Patients in the second category are designated as having type C NPD. Impaired intracellular trafficking of cholesterol causes type C NPD, and two distinct gene defects have been found. In this chapter only types A and B NPD will be discussed. PMID:27491215

  7. SMPD1 Mutation Update: Database and Comprehensive Analysis of Published and Novel Variants.

    PubMed

    Zampieri, Stefania; Filocamo, Mirella; Pianta, Annalisa; Lualdi, Susanna; Gort, Laura; Coll, Maria Jose; Sinnott, Richard; Geberhiwot, Tarekegn; Bembi, Bruno; Dardis, Andrea

    2016-02-01

    Niemann-Pick Types A and B (NPA/B) diseases are autosomal recessive lysosomal storage disorders caused by the deficient activity of acid sphingomyelinase (ASM) because of the mutations in the SMPD1 gene. Here, we provide a comprehensive updated review of already reported and newly identified SMPD1 variants. Among them, 185 have been found in NPA/B patients. Disease-causing variants are equally distributed along the SMPD1 gene; most of them are missense (65.4%) or frameshift (19%) mutations. The most frequently reported mutation worldwide is the p.R610del, clearly associated with an attenuated NP disease type B phenotype. The available information about the impact of 52 SMPD1 variants on ASM mRNA and/or enzymatic activity has been collected and whenever possible, phenotype/genotype correlations were established. In addition, we created a locus-specific database easily accessible at http://www.inpdr.org/genes that catalogs the 417 SMPD1 variants reported to date and provides data on their in silico predicted effects on ASM protein function or mRNA splicing. The information reviewed in this article, providing new insights into the genotype/phenotype correlation, is extremely valuable to facilitate diagnosis and genetic counseling of families affected by NPA/B. PMID:26499107

  8. Brown recluse spider (Loxosceles reclusa) venom phospholipase D (PLD) generates lysophosphatidic acid (LPA)

    PubMed Central

    2005-01-01

    Envenomation by the brown recluse spider (Loxosceles reclusa) may cause local dermonecrosis and, rarely, coagulopathies, kidney failure and death. A venom phospholipase, SMaseD (sphingomyelinase D), is responsible for the pathological manifestations of envenomation. Recently, the recombinant SMaseD from Loxosceles laeta was demonstrated to hydrolyse LPC (lysophosphatidylcholine) to produce LPA (lysophosphatidic acid) and choline. Therefore activation of LPA signalling pathways may be involved in some manifestations of Loxosceles envenomation. To begin investigating this idea, we cloned a full-length cDNA encoding L. reclusa SMaseD. The 305 amino acid sequence of the L. reclusa enzyme is 87, 85 and 60% identical with those of L. arizonica, L. intermedia and L. laeta respectively. The recombinant enzyme expressed in bacteria had broad substrate specificity. The lysophospholipids LPC, LPI (18:1-1-oleyol lysophosphatidylinositol), LPS, LPG (18:1-1-oleoyl-lysophosphatidylglycerol), LBPA (18:1-1-oleoyl-lysobisphosphatidic acid) (all with various acyl chains), lyso-platelet-activating factor (C16:0), cyclic phosphatidic acid and sphingomyelin were hydrolysed, whereas sphingosylphosphorylcholine, PC (phosphatidylcholine; C22:6, C20:4 and C6:0), oxidized PCs and PAF (platelet-activating factor; C16:0) were not hydrolysed. The PAF analogue, edelfosine, inhibited enzyme activity. Recombinant enzyme plus LPC (C18:1) induced the migration of A2058 melanoma cells, and this activity was blocked by the LPA receptor antagonist, VPC32183. The recombinant spider enzyme was haemolytic, but this activity was absent from catalytically inactive H37N (His37→Asn) and H73N mutants. Our results demonstrate that Loxosceles phospholipase D hydrolyses a wider range of lysophospholipids than previously supposed, and thus the term ‘SMaseD’ is too limited in describing this enzyme. PMID:15926888

  9. Brown recluse spider (Loxosceles reclusa) venom phospholipase D (PLD) generates lysophosphatidic acid (LPA).

    PubMed

    Lee, Sangderk; Lynch, Kevin R

    2005-10-15

    Envenomation by the brown recluse spider (Loxosceles reclusa) may cause local dermonecrosis and, rarely, coagulopathies, kidney failure and death. A venom phospholipase, SMaseD (sphingomyelinase D), is responsible for the pathological manifestations of envenomation. Recently, the recombinant SMaseD from Loxosceles laeta was demonstrated to hydrolyse LPC (lysophosphatidylcholine) to produce LPA (lysophosphatidic acid) and choline. Therefore activation of LPA signalling pathways may be involved in some manifestations of Loxosceles envenomation. To begin investigating this idea, we cloned a full-length cDNA encoding L. reclusa SMaseD. The 305 amino acid sequence of the L. reclusa enzyme is 87, 85 and 60% identical with those of L. arizonica, L. intermedia and L. laeta respectively. The recombinant enzyme expressed in bacteria had broad substrate specificity. The lysophospholipids LPC, LPI (18:1-1-oleyol lysophosphatidylinositol), LPS, LPG (18:1-1-oleoyl-lysophosphatidylglycerol), LBPA (18:1-1-oleoyl-lysobisphosphatidic acid) (all with various acyl chains), lyso-platelet-activating factor (C16:0), cyclic phosphatidic acid and sphingomyelin were hydrolysed, whereas sphingosylphosphorylcholine, PC (phosphatidylcholine; C22:6, C20:4 and C6:0), oxidized PCs and PAF (platelet-activating factor; C16:0) were not hydrolysed. The PAF analogue, edelfosine, inhibited enzyme activity. Recombinant enzyme plus LPC (C18:1) induced the migration of A2058 melanoma cells, and this activity was blocked by the LPA receptor antagonist, VPC32183. The recombinant spider enzyme was haemolytic, but this activity was absent from catalytically inactive H37N (His37-->Asn) and H73N mutants. Our results demonstrate that Loxosceles phospholipase D hydrolyses a wider range of lysophospholipids than previously supposed, and thus the term 'SMaseD' is too limited in describing this enzyme. PMID:15926888

  10. Report on task assignment No. 3 for the Waste Package Project; Parts A & B, ASME pressure vessel codes review for waste package application; Part C, Library search for reliability/failure rates data on low temperature low pressure piping, containers, and casks with long design lives

    SciTech Connect

    Trabia, M.B.; Kiley, M.; Cardle, J.; Joseph, M.

    1991-07-01

    The Waste Package Project Research Team, at UNLV, has four general required tasks. Task one is the management, quality assurance, and overview of the research that is performed under the cooperative agreement. Task two is the structural analysis of spent fuel and high level waste. Task three is an American Society of Mechanical Engineers (ASME) Pressure Vessel Code review for waste package application. Finally, task four is waste package labeling. This report includes preliminary information about task three (ASME Pressure Vessel Code review for Waste package Application). The first objective is to compile a list of the ASME Pressure Vessel Code that can be applied to waste package containers design and manufacturing processes. The second objective is to explore the use of these applicable codes to the preliminary waste package container designs. The final objective is to perform a library search for reliability and/or failure rates data on low pressure, low temperature, containers and casks with long design lives.

  11. Biomineralization of arsenate to arsenic sulfides is greatly enhanced at mildly acidic conditions.

    PubMed

    Rodriguez-Freire, Lucia; Sierra-Alvarez, Reyes; Root, Robert; Chorover, Jon; Field, James A

    2014-12-01

    Arsenic (As) is an important water contaminant due to its high toxicity and widespread occurrence. Arsenic-sulfide minerals (ASM) are formed during microbial reduction of arsenate (As(V)) and sulfate (SO4(2-)). The objective of this research is to study the effect of the pH on the removal of As due to the formation of ASM in an iron-poor system. A series of batch experiments was used to study the reduction of SO4(2-) and As(V) by an anaerobic biofilm mixed culture in a range of pH conditions (6.1-7.2), using ethanol as the electron donor. Total soluble concentrations and speciation of S and As were monitored. Solid phase speciation of arsenic was characterized by x-ray adsorption spectroscopy (XAS). A marked decrease of the total aqueous concentrations of As and S was observed in the inoculated treatments amended with ethanol, but not in the non-inoculated controls, indicating that the As-removal was biologically mediated. The pH dramatically affected the extent and rate of As removal, as well as the stoichiometric composition of the precipitate. The amount of As removed was 2-fold higher and the rate of the As removal was up to 17-fold greater at pH 6.1 than at pH 7.2. Stoichiometric analysis and XAS results confirmed the precipitate was composed of a mixture of orpiment and realgar, and the proportion of orpiment in the sample increased with increasing pH. The results taken as a whole suggest that ASM formation is greatly enhanced at mildly acidic pH conditions. PMID:25222328

  12. Biomineralization of Arsenate to Arsenic Sulfides is Greatly Enhanced at Mildly Acidic Conditions

    PubMed Central

    Rodriguez-Freire, Lucia; Sierra-Alvarez, Reyes; Root, Robert; Chorover, Jon; Field, James A.

    2014-01-01

    Arsenic (As) is an important water contaminant due to its high toxicity and widespread occurrence. Arsenic-sulfide minerals (ASM) are formed during microbial reduction of arsenate (AsV) and sulfate (SO42−). The objective of this research is to study the effect of the pH on the removal of As due to the formation of ASM in an iron-poor system. A series of batch experiments was used to study the reduction of SO42− and AsV by an anaerobic biofilm mixed culture in a range of pH conditions (6.1–7.2), using ethanol as the electron donor. Total soluble concentrations and speciation of S and As were monitored. Solid phase speciation of arsenic was characterized by x-ray adsorption spectroscopy (XAS). A marked decrease of the total aqueous concentrations of As and S was observed in the inoculated treatments amended with ethanol, but not in the non-inoculated controls, indicating that the As-removal was biologically mediated. The pH dramatically affected the extent and rate of As removal, as well as the stoichiometric composition of the precipitate. The amount of As removed was 2-fold higher and the rate of the As removal was up to 17-fold greater at pH 6.1 than at pH 7.2. Stoichiometric analysis and XAS results confirmed the precipitate was composed of a mixture of orpiment and realgar, and the proportion of orpiment in the sample increased with increasing pH. The results taken as a whole suggest that ASM formation is greatly enhanced at mildly acidic pH conditions. PMID:25222328

  13. Amino acids

    MedlinePlus

    ... this page: //medlineplus.gov/ency/article/002222.htm Amino acids To use the sharing features on this page, please enable JavaScript. Amino acids are organic compounds that combine to form proteins . ...

  14. Mefenamic Acid

    MedlinePlus

    Mefenamic acid is used to relieve mild to moderate pain, including menstrual pain (pain that happens before or during a menstrual period). Mefenamic acid is in a class of medications called NSAIDs. ...

  15. Aminocaproic Acid

    MedlinePlus

    Aminocaproic acid is used to control bleeding that occurs when blood clots are broken down too quickly. This type ... the baby is ready to be born). Aminocaproic acid is also used to control bleeding in the ...

  16. Ascorbic Acid

    MedlinePlus

    Ascorbic acid is used to prevent and treat scurvy, a disease caused by a lack of vitamin C in ... Ascorbic acid comes in extended-release (long-acting) capsules and tablets, lozenges, syrup, chewable tablets, and liquid drops to ...

  17. Acid mucopolysaccharides

    MedlinePlus

    ... this page: //medlineplus.gov/ency/article/003368.htm Acid mucopolysaccharides To use the sharing features on this page, please enable JavaScript. Acid mucopolysaccharides is a test that measures the amount ...

  18. Ethacrynic Acid

    MedlinePlus

    Ethacrynic acid, a 'water pill,' is used to treat swelling and fluid retention caused by various medical problems. It ... Ethacrynic acid comes as a tablet to take by mouth. It is usually taken once or twice a day ...

  19. EDITORIAL: Adaptive and Active Materials: Selected Papers from the ASME 2008 Conference on Smart Materials, Adaptive Structures and Intelligent Systems (SMASIS 08) (Maryland, USA, 28-30 October 2008) Adaptive and Active Materials: Selected Papers from the ASME 2008 Conference on Smart Materials, Adaptive Structures and Intelligent Systems (SMASIS 08) (Maryland, USA, 28-30 October 2008)

    NASA Astrophysics Data System (ADS)

    Lynch, Christopher

    2009-10-01

    The rapid development of the field of Smart Materials, Adaptive Structures, and Materials Systems led the Aerospace Division ASMS TC to launch the new annual SMASIS conference in 2008. The conference focuses on the multi-disciplinary challenges of developing new multifunctional materials and implementing them in advanced systems. The research spans length scales from nano-structured materials to civil, air, and space structures. The first conference consisted of six symposia, each focusing on a different research area. This special issue of Smart Materials and Structures summarizes some of the top research presented at the 2008 SMASIS conference in the materials-focused symposia. These symposia focused on the behavior and mechanics of active materials, on multifunctional materials, and on bio-inspired materials. The behavior and mechanics of active materials is an approach that combines observed material behavior with mechanism-based models that not only give insight into the observed behavior, but guide the development of new materials. This approach has been applied to shape memory metals and polymers, ferroelectrics, ferromagnetics, and recently to multiferroic materials, and has led to considerable improvements in our understanding of multi-field phenomena. Multifunctional materials are the next generation of active materials. These materials include structural, sensing, and actuation components integrated into a material system. A natural extension of multifunctional materials is a new class of bio-inspired materials. Bio-inspired materials range from detailed bio-mimicry of sensing and self healing materials to nano and microstructures that take advantage of features observed in biological systems. The Editors would like to express their sincere thanks to all of the authors for their contributions to this special issue on 'Adaptive and Active Materials' for Smart Materials and Structures. We convey our gratitude to all of the reviewers for their time and

  20. Valproic Acid

    MedlinePlus

    Valproic acid is used alone or with other medications to treat certain types of seizures. Valproic acid is also used to treat mania (episodes of ... to relieve headaches that have already begun. Valproic acid is in a class of medications called anticonvulsants. ...

  1. Fatty acids - trans fatty acids

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The data supporting a negative effect of dietary trans fatty acids on cardiovascular disease risk is consistent. The primary dietary sources of trans fatty acids include partially hydrogenated fat and rudiment fat. The adverse effect of trans fatty acids on plasma lipoprotein profiles is consisten...

  2. In Vitro and In Vivo Activities of Arachidonic Acid against Schistosoma mansoni and Schistosoma haematobium▿

    PubMed Central

    El Ridi, Rashika; Aboueldahab, Marwa; Tallima, Hatem; Salah, Mohamed; Mahana, Noha; Fawzi, Samia; Mohamed, Shadia H.; Fahmy, Omar M.

    2010-01-01

    The development of arachidonic acid (ARA) for treatment of schistosomiasis is an entirely novel approach based on a breakthrough discovery in schistosome biology revealing that activation of parasite tegument-bound neutral sphingomyelinase (nSMase) by unsaturated fatty acids, such as ARA, induces exposure of parasite surface membrane antigens to antibody binding and eventual attrition of developing schistosomula and adult worms. Here, we demonstrate that 5 mM ARA leads to irreversible killing of ex vivo 1-, 3-, 4-, 5-, and 6-week-old Schistosoma mansoni and 9-, 10-, and 12-week-old Schistosoma haematobium worms within 3 to 4 h, depending on the parasite age, even when the worms were maintained in up to 50% fetal calf serum. ARA-mediated worm attrition was prevented by nSMase inhibitors, such as CaCl2 and GW4869. Scanning and transmission electron microscopy revealed that ARA-mediated worm killing was associated with spine destruction, membrane blebbing, and disorganization of the apical membrane structure. ARA-mediated S. mansoni and S. haematobium worm attrition was reproduced in vivo in a series of 6 independent experiments using BALB/c or C57BL/6 mice, indicating that ARA in a pure form (Sigma) or included in infant formula (Nestle) consistently led to 40 to 80% decrease in the total worm burden. Arachidonic acid is already marketed for human use in the United States and Canada for proper development of newborns and muscle growth of athletes; thus, ARA has potential as a safe and cost-effective addition to antischistosomal therapy. PMID:20479203

  3. Acid Deposition

    EPA Science Inventory

    This indicator presents acid deposition trends in the contiguous U.S. from 1989 to 2007. Data are broken down by wet and dry deposition and deposition of nitrogen and sulfur compounds. Acid deposition is particularly damaging to lakes, streams, and forests and the plants and a...

  4. Acid rain

    SciTech Connect

    White, J.C. )

    1988-01-01

    This book presents the proceedings of the third annual conference sponsored by the Acid Rain Information Clearinghouse (ARIC). Topics covered include: Legal aspects of the source-receptor relationship: an energy perspective; Scientific uncertainty, agency inaction, and the courts; and Acid rain: the emerging legal framework.

  5. Acid rain

    SciTech Connect

    Elsworth, S.

    1985-01-01

    This book was written in a concise and readable style for the lay public. It's purpose was to make the public aware of the damage caused by acid rain and to mobilize public opinion to favor the elimination of the causes of acid rain.

  6. Acid rain

    SciTech Connect

    Sweet, W.

    1980-06-20

    Acid precipitation includes not only rain but also acidified snow, hail and frost, as well as sulfur and nitrogen dust. The principal source of acid precipitation is pollution emitted by power plants and smelters. Sulfur and nitrogen compounds contained in the emissions combine with moisture to form droplets with a high acid content - sometimes as acidic as vinegar. When sufficiently concentrated, these acids can kill fish and damage material structures. Under certain circumstances they may reduce crop and forest yields and cause or aggravate respiratory diseases in humans. During the summer, especially, pollutants tend to collect over the Great Lakes in high pressure systems. Since winds typically are westerly and rotate clockwise around high pressure systems, the pollutants gradually are dispersed throughout the eastern part of the continent.

  7. Asparagusic acid.

    PubMed

    Mitchell, Stephen C; Waring, Rosemary H

    2014-01-01

    Asparagusic acid (1,2-dithiolane-4-carboxylic acid) is a simple sulphur-containing 5-membered heterocyclic compound that appears unique to asparagus, though other dithiolane derivatives have been identified in non-food species. This molecule, apparently innocuous toxicologically to man, is the most probable culprit responsible for the curious excretion of odorous urine following asparagus ingestion. The presence of the two adjacent sulphur atoms leads to an enhanced chemical reactivity, endowing it with biological properties including the ability to substitute potentially for α-lipoic acid in α-keto-acid oxidation systems. This brief review collects the scattered data available in the literature concerning asparagusic acid and highlights its properties, intermediary metabolism and exploratory applications. PMID:24099657

  8. [Gastric Acid].

    PubMed

    Ruíz Chávez, R

    1996-01-01

    Gastric acid, a product of parietal cells secretion, full fills multiple biological roles which are absolutely necessary to keep corporal homeostasis. The production of the acid depends upon an effector cellular process represented in the first step by histamine, acetilcholine and gastrin, first messengers of the process. These interact with specific receptors than in sequence activate second messengers -cAMP and the calcium-calmodulin system- which afterwards activate a kinase. An specific protein is then phosphorilated by this enzyme, being the crucial factor that starts the production of acid. Finally, a proton bomb, extrudes the acid towards the gastric lumen. The secretion process mentioned above, is progressive lyactivated in three steps, two of which are stimulators -cephalic and gastric phases- and the other one inhibitor or intestinal phase. These stages are started by mental and neurological phenomena -thought, sight, smell or memory-; by food, drugs or other ingested substances; and by products of digestion. Changes in regulation of acid secretion, in the structure of gastro-duodenal mucosal barrier by a wide spectrum of factors and agents including food, drugs and H. pylori, are the basis of acid-peptic disease, entity in which gastric acid plays a fundamental role. From the therapeutic point of view, so at the theoretical as at the practical levels, t is possible to interfere with the secretion of acid by neutralization of some of the steps of the effector cellular process. An adequate knowledge of the basics related to gastric acid, allows to create strategies for the clinical handling of associated pathology, specifically in relation to peptic acid disease in all of the known clinical forms. PMID:12165790

  9. Ursodeoxycholic acid upregulates ERK and Akt in the protection of cardiomyocytes against CoCl2.

    PubMed

    Hanafi, N I; Mohamed, A S; Md Noor, J; Abdu, N; Hasani, Hamid; Siran, R; Osman, N J; Ab Rahim, S; Sheikh Abdul Kadir, S H

    2016-01-01

    Ursodeoxycholic acid (UDCA) is used to treat liver diseases and demonstrates cardioprotective effects. Accumulation of the plasma membrane sphingolipid sphingomyelin in the heart can lead to atherosclerosis and coronary artery disease. Sphingomyelinases (SMases) break down sphingomyelin, producing ceramide, and inhibition of SMases activity can promote cell survival. We hypothesized that UDCA regulates activation of ERK and Akt survival signaling pathways and SMases in protecting cardiac cells against hypoxia. Neonatal cardiomyocytes were isolated from 0- to 2-day-old Sprague Dawley rats, and given 100 μM CoCl2, 150 μM H2O2, or placed in a hypoxia chamber for 24 h. The ameliorative effects of 100-μM UDCA treatment for 12 h were then assessed using MTS, QuantiGene Plex (for Smpd1 and Smpd2), and SMase assays, beating rate assessment, and western blotting (for ERK and Akt). Data were analyzed by the paired Student t-tests and one-way analyses of variance. Cell viability decreased significantly after H2O2 (85%), CoCl2 (50%), and hypoxia chamber (52%) treatments compared to the untreated control (100%). UDCA significantly counteracted the effects of chamber- and CoCl2- induced hypoxia on viability and beating rate. However, no significant differences were observed in acid SMase gene and protein expression between the untreated, CoCl2, and UDCA-CoCl2 groups. In contrast, neutral SMase gene and protein expression did significantly differ between the latter two groups. ERK and Akt phosphorylation was higher in hypoxic cardiomyocytes treated with UDCA than those given CoCl2 alone. In conclusion, UDCA regulates the activation of survival signaling proteins and SMases in neonatal rat cardiomyocytes during hypoxia. PMID:27323195

  10. Acid fog

    SciTech Connect

    Hileman, B.

    1983-03-01

    Fog in areas of southern California previously thought to be pollution-free has been shown to have a pH as low as 1.69. It has been found to be most acidic after smoggy days, suggesting that it forms on the aerosol associated with the previously exiting smog. Studies on Whiteface Mountain in the Adirondacks show that fog water is often 10 times as acidic as rainwater. As a result of their studies, California plans to spend $4 million on acid deposition research in the coming year. (JMT)

  11. Folic acid

    MedlinePlus

    ... in the blood vessel to keep it open. Bipolar disorder. Taking folic acid does not appear to improve the antidepressant effects of lithium in people with bipolar disorder. However, taking folate with the medication valproate improves ...

  12. Mefenamic Acid

    MedlinePlus

    ... as mefenamic acid may cause ulcers, bleeding, or holes in the stomach or intestine. These problems may ... like coffee grounds, blood in the stool, or black and tarry stools.Keep all appointments with your ...

  13. ACID RAIN

    EPA Science Inventory

    Acid precipitation has become one of the major environmental problems of this decade. It is a challenge to scientists throughout the world. Researchers from such diverse disciplines as plant pathology, soil science, bacteriology, meteorology and engineering are investigating diff...

  14. Acid Precipitation

    ERIC Educational Resources Information Center

    Likens, Gene E.

    1976-01-01

    Discusses the fact that the acidity of rain and snow falling on parts of the U.S. and Europe has been rising. The reasons are still not entirely clear and the consequences have yet to be well evaluated. (MLH)

  15. Carnosic acid.

    PubMed

    Birtić, Simona; Dussort, Pierre; Pierre, François-Xavier; Bily, Antoine C; Roller, Marc

    2015-07-01

    Carnosic acid (salvin), which possesses antioxidative and antimicrobial properties, is increasingly exploited within the food, nutritional health and cosmetics industries. Since its first extraction from a Salvia species (∼70 years ago) and its identification (∼50 years ago), numerous articles and patents (∼400) have been published on specific food and medicinal applications of Rosmarinus and Salvia plant extracts abundant in carnosic acid. In contrast, relevant biochemical, physiological or molecular studies in planta have remained rare. In this overview, recent advances in understanding of carnosic acid distribution, biosynthesis, accumulation and role in planta, and its applications are summarised. We also discuss the deficiencies in our understanding of the relevant biochemical processes, and suggest the molecular targets of carnosic acid. Finally, future perspectives and studies related to its potential roles are highlighted. PMID:25639596

  16. Aminocaproic Acid

    MedlinePlus

    Amicar® Oral Solution ... Aminocaproic acid comes as a tablet and a solution (liquid) to take by mouth. It is usually ... it at room temperature and away from excess heat and moisture (not in the bathroom). Throw away ...

  17. Tranexamic Acid

    MedlinePlus

    ... is used to treat heavy bleeding during the menstrual cycle (monthly periods) in women. Tranexamic acid is in ... tablets for more than 5 days in a menstrual cycle or take more than 6 tablets in a ...

  18. EDITORIAL: Adaptive and active materials: Selected papers from the ASME 2011 Conference on Smart Materials, Adaptive Structures and Intelligent Systems (SMASIS 11) (Scottsdale, AZ, USA, 18-21 September 2011) Adaptive and active materials: Selected papers from the ASME 2011 Conference on Smart Materials, Adaptive Structures and Intelligent Systems (SMASIS 11) (Scottsdale, AZ, USA, 18-21 September 2011)

    NASA Astrophysics Data System (ADS)

    Brei, Diann

    2012-09-01

    The fourth annual meeting of the ASME/AIAA Smart Materials, Adaptive Structures and Intelligent Systems Conference (SMASIS) took place in sunny Scottsdale, Arizona. Each year we strive to grow and offer new experiences. This year we held a special Guest Symposium on Sustainability along with two focused topic tracks on energy harvesting and active composites to encourage cross-fertilization between these important fields and our community. This cross-disciplinary emphasis was reflected in keynote talks by Dr Wayne Brown, President and founder of Dynalloy, Inc., 'Cross-Discipline Sharing'; Dr Brad Allenby, Arizona State University, 'You Want the Future? You can't Handle the Future!'; and Professor Aditi Chattopadhyay, Arizona State University, 'A Multidisciplinary Approach to Structural Health Monitoring and Prognosis'. SMASIS continues to grow our community through both social and technical interchange. The conference location, the exotic Firesky Resort and Spa, exemplified the theme of our Guest Symposium on Sustainability, being the only Green Seal certified resort in Arizona, and highlighting four elements thought to represent all that exist: fire, water, earth and air. Several special events were held around this theme including the night at the oasis reception sponsored by General Motors, sustainability bingo, smart trivia and student networking lunches, and an Arizona pow-wow with a spectacular Indian hoop dance. Our student and young professional development continues to grow strong with best paper and hardware competitions, scavenger student outing and games night. We are very proud that our students and young professionals are always seeking out ways to give back to the community, including organizing outreach to local high school talent. We thank all of our sponsors who made these special events possible. We hope that these social events provided participants with the opportunity to expand their own personal community and broaden their horizons. Our

  19. Acidic precipitation

    SciTech Connect

    Martin, H.C.

    1987-01-01

    At the International Symposium on Acidic Precipitation, over 400 papers were presented, and nearly 200 of them are included here. They provide an overview of the present state of the art of acid rain research. The Conference focused on atmospheric science (monitoring, source-receptor relationships), aquatic effects (marine eutrophication, lake acidification, impacts on plant and fish populations), and terrestrial effects (forest decline, soil acidification, etc.).

  20. Synthesis, structure and properties of the oxychalcogenide series A{sub 4}O{sub 4}TiSe{sub 4} (A=Sm, Gd, Tb, Dy, Ho, Er and Y)

    SciTech Connect

    Tuxworth, A.J.; Evans, J.S.O.

    2014-02-15

    Seven oxyselenide materials have been synthesised with composition A{sub 4}O{sub 4}TiSe{sub 4} (A=Sm, Gd–Er, Y) via solid state reactions of A{sub 2}O{sub 3}, TiSe{sub 2} and Se at 900 °C. They are all isostructural with Gd{sub 4}O{sub 4}TiSe{sub 4}. Structures have been refined from powder X-ray diffraction data and have monoclinic C2/m symmetry with unit cell parameters of a≈15.7 Å, b≈3.75 Å, c≈9.65 Å and β≈117.5°. They contain infinite ribbons of edge-sharing A{sub 4}O and A{sub 3}TiO tetrahedra 4 units wide, which are linked by chains of TiSe{sub 4}O{sub 2} edge-sharing octahedra. Compositions A=Gd–Ho, Y are semiconductors with conductivities 1–3 Sm{sup −1} at 300 K, with electronic band gaps of between 0.25 and 0.37 eV. Magnetic susceptibility is reported from 1.8 K to 300 K for compositions A=Gd–Ho. Rare earth moments appear to order antiferromagnetically at low temperatures with Gd and Tb showing evidence of ferromagnetism due to spin canting over a narrow temperature range close to T{sub N}. - Graphical abstract: Illustration of the A{sub 4}O{sub 4}TiSe{sub 4} crystal structure (C2/m symmetry), A{sub 4}O and A{sub 3}TiO edge sharing tetrahedral ribbons in red, chains of edge-sharing TiSe{sub 4}O{sub 2} in blue. Rare earth=green, titanium=blue, selenium=yellow, and oxygen=red. Display Omitted - Highlights: • Seven materials with A{sub 4}O{sub 4}TiSe{sub 4} (A=Sm, Gd–Er, Y) have been synthesised as bulk phases for the first time. • Materials are semiconductors and order antiferromagnetically at ∼4 K. • Structure contains M{sub 4}O tetrahedral ribbons and TiSe{sub 4}O{sub 2} 1D octahedral chains.

  1. Salicylic acids

    PubMed Central

    Hayat, Shamsul; Irfan, Mohd; Wani, Arif; Nasser, Alyemeni; Ahmad, Aqil

    2012-01-01

    Salicylic acid is well known phytohormone, emerging recently as a new paradigm of an array of manifestations of growth regulators. The area unleashed yet encompassed the applied agriculture sector to find the roles to strengthen the crops against plethora of abiotic and biotic stresses. The skipped part of integrated picture, however, was the evolutionary insight of salicylic acid to either allow or discard the microbial invasion depending upon various internal factors of two interactants under the prevailing external conditions. The metabolic status that allows the host invasion either as pathogenesis or symbiosis with possible intermediary stages in close systems has been tried to underpin here. PMID:22301975

  2. Folic acid

    MedlinePlus

    ... disease called vitiligo, and an inherited disease called Fragile-X syndrome. It is also used for reducing harmful side ... to blood clots (ischemic stroke). Inherited disease called Fragile-X syndrome.Taking folic acid by mouth does not improve ...

  3. Acid rain

    SciTech Connect

    Not Available

    1984-06-01

    An overview is presented of acid rain and the problems it causes to the environment worldwide. The acidification of lakes and streams is having a dramatic effect on aquatic life. Aluminum, present in virtually all forest soils, leaches out readily under acid conditions and interferes with the gills of all fish, some more seriously than others. There is evidence of major damage to forests in European countries. In the US, the most severe forest damage appears to be in New England, New York's Adirondacks, and the central Appalachians. This small region is part of a larger area of the Northeast and Canada that appears to have more acid rainfall than the rest of the country. It is downwind from major coal burning states, which produce about one quarter of US SO/sub 2/ emissions and one sixth of nitrogen oxide emissions. Uncertainties exist over the causes of forest damage and more research is needed before advocating expensive programs to reduce rain acidity. The President's current budget seeks an expansion of research funds from the current $30 million per year to $120 million.

  4. Formic acid

    Integrated Risk Information System (IRIS)

    Formic acid ; CASRN 64 - 18 - 6 Human health assessment information on a chemical substance is included in the IRIS database only after a comprehensive review of toxicity data , as outlined in the IRIS assessment development process . Sections I ( Health Hazard Assessments for Noncarcinogenic Effect

  5. Selenious acid

    Integrated Risk Information System (IRIS)

    Selenious acid ; CASRN 7783 - 00 - 8 Human health assessment information on a chemical substance is included in the IRIS database only after a comprehensive review of toxicity data , as outlined in the IRIS assessment development process . Sections I ( Health Hazard Assessments for Noncarcinogenic E

  6. Benzoic acid

    Integrated Risk Information System (IRIS)

    Benzoic acid ; CASRN 65 - 85 - 0 Human health assessment information on a chemical substance is included in the IRIS database only after a comprehensive review of toxicity data , as outlined in the IRIS assessment development process . Sections I ( Health Hazard Assessments for Noncarcinogenic Effec

  7. Trichloroacetic acid

    Integrated Risk Information System (IRIS)

    Trichloroacetic acid ( TCA ) ; CASRN 76 - 03 - 9 Human health assessment information on a chemical substance is included in the IRIS database only after a comprehensive review of toxicity data , as outlined in the IRIS assessment development process . Sections I ( Health Hazard Assessments for Nonca

  8. Dichloroacetic acid

    Integrated Risk Information System (IRIS)

    Dichloroacetic acid ; CASRN 79 - 43 - 6 Human health assessment information on a chemical substance is included in the IRIS database only after a comprehensive review of toxicity data , as outlined in the IRIS assessment development process . Sections I ( Health Hazard Assessments for Noncarcinogeni

  9. Acrylic acid

    Integrated Risk Information System (IRIS)

    Acrylic acid ( CASRN 79 - 10 - 7 ) Human health assessment information on a chemical substance is included in the IRIS database only after a comprehensive review of toxicity data , as outlined in the IRIS assessment development process . Sections I ( Health Hazard Assessments for Noncarcinogenic Eff

  10. Cacodylic acid

    Integrated Risk Information System (IRIS)

    Cacodylic acid ; CASRN 75 - 60 - 5 Human health assessment information on a chemical substance is included in the IRIS database only after a comprehensive review of toxicity data , as outlined in the IRIS assessment development process . Sections I ( Health Hazard Assessments for Noncarcinogenic Eff

  11. Phosphoric acid

    Integrated Risk Information System (IRIS)

    Phosphoric acid ; CASRN 7664 - 38 - 2 Human health assessment information on a chemical substance is included in the IRIS database only after a comprehensive review of toxicity data , as outlined in the IRIS assessment development process . Sections I ( Health Hazard Assessments for Noncarcinogenic

  12. Stearic Acid

    ERIC Educational Resources Information Center

    Young, Jay A.

    2004-01-01

    A chemical laboratory information profile (CLIP) is presented for the chemical, stearic acid. The profile lists the chemical's physical and harmful characteristics, exposure limits, and symptoms of major exposure, for the benefit of teachers and students, who use the chemical in the laboratory.

  13. Host sphingomyelin increases West Nile virus infection in vivo.

    PubMed

    Martín-Acebes, Miguel A; Gabandé-Rodríguez, Enrique; García-Cabrero, Ana M; Sánchez, Marina P; Ledesma, María Dolores; Sobrino, Francisco; Saiz, Juan-Carlos

    2016-03-01

    Flaviviruses, such as the dengue virus and the West Nile virus (WNV), are arthropod-borne viruses that represent a global health problem. The flavivirus lifecycle is intimately connected to cellular lipids. Among the lipids co-opted by flaviviruses, we have focused on SM, an important component of cellular membranes particularly enriched in the nervous system. After infection with the neurotropic WNV, mice deficient in acid sphingomyelinase (ASM), which accumulate high levels of SM in their tissues, displayed exacerbated infection. In addition, WNV multiplication was enhanced in cells from human patients with Niemann-Pick type A, a disease caused by a deficiency of ASM activity resulting in SM accumulation. Furthermore, the addition of SM to cultured cells also increased WNV infection, whereas treatment with pharmacological inhibitors of SM synthesis reduced WNV infection. Confocal microscopy analyses confirmed the association of SM with viral replication sites within infected cells. Our results unveil that SM metabolism regulates flavivirus infection in vivo and propose SM as a suitable target for antiviral design against WNV. PMID:26764042

  14. Induction of Membrane Ceramides: A Novel Strategy to Interfere with T Lymphocyte Cytoskeletal Reorganisation in Viral Immunosuppression

    PubMed Central

    Harms, Harry; Krohne, Georg; Gulbins, Erich; Schneider-Schaulies, Sibylle

    2009-01-01

    Silencing of T cell activation and function is a highly efficient strategy of immunosuppression induced by pathogens. By promoting formation of membrane microdomains essential for clustering of receptors and signalling platforms in the plasma membrane, ceramides accumulating as a result of membrane sphingomyelin breakdown are not only essential for assembly of signalling complexes and pathogen entry, but also act as signalling modulators, e. g. by regulating relay of phosphatidyl-inositol-3-kinase (PI3K) signalling. Their role in T lymphocyte functions has not been addressed as yet. We now show that measles virus (MV), which interacts with the surface of T cells and thereby efficiently interferes with stimulated dynamic reorganisation of their actin cytoskeleton, causes ceramide accumulation in human T cells in a neutral (NSM) and acid (ASM) sphingomyelinase–dependent manner. Ceramides induced by MV, but also bacterial sphingomyelinase, efficiently interfered with formation of membrane protrusions and T cell spreading and front/rear polarisation in response to β1 integrin ligation or αCD3/CD28 activation, and this was rescued upon pharmacological or genetic ablation of ASM/NSM activity. Moreover, membrane ceramide accumulation downmodulated chemokine-induced T cell motility on fibronectin. Altogether, these findings highlight an as yet unrecognised concept of pathogens able to cause membrane ceramide accumulation to target essential processes in T cell activation and function by preventing stimulated actin cytoskeletal dynamics. PMID:19834551

  15. Hydroxycarboxylic acids and salts

    DOEpatents

    Kiely, Donald E; Hash, Kirk R; Kramer-Presta, Kylie; Smith, Tyler N

    2015-02-24

    Compositions which inhibit corrosion and alter the physical properties of concrete (admixtures) are prepared from salt mixtures of hydroxycarboxylic acids, carboxylic acids, and nitric acid. The salt mixtures are prepared by neutralizing acid product mixtures from the oxidation of polyols using nitric acid and oxygen as the oxidizing agents. Nitric acid is removed from the hydroxycarboxylic acids by evaporation and diffusion dialysis.

  16. Adaptive and active materials: selected papers from the ASME 2013 Conference on Smart Materials, Adaptive Structures and Intelligent Systems (SMASIS 13) (Snowbird, UT, USA, 16-18 September 2013)

    NASA Astrophysics Data System (ADS)

    Johnson, Nancy; Naguib, Hani; Turner, Travis; Anderson, Iain; Bassiri-Gharb, Nazanin; Daqaq, Mohammed; Baba Sundaresan, Vishnu; Sarles, Andy

    2014-10-01

    The sixth annual meeting of the ASME Smart Materials, Adaptive Structures and Intelligent Systems Conference (SMASIS) was held in the beautiful mountain encircled Snowbird Resort and Conference Center in Little Cottonwood Canyon near Salt Lake City, Utah. It is the conference's objective to provide an up-to-date overview of research trends in the entire field of smart materials systems in a friendly casual forum conducive to the exchange of ideas and latest results. As each year we strive to grow and offer new experiences, this year we included special focused topic tracks on nanoscale multiferroic materials and origami engineering. The cross-disciplinary emphasis was reflected in keynote speeches by Professor Kaushik Bhattacharya (California Institute of Technology) on 'Cyclic Deformation and the Interplay between Phase Transformation and Plasticity in Shape Memory Alloys', by Professor Alison Flatau (University of Maryland at College Park) on 'Structural Magnetostrictive Alloys: The Other Smart Material', and by Dr Leslie Momoda (Director of the Sensors and Materials Laboratories, HRL Laboratories, LLC, Malibu, CA) on 'Architecturing New Functional Materials: An Industrial Perspective'. SMASIS 2013 was divided into seven symposia which span basic research, applied technological design and development, and industrial and governmental integrated system and application demonstrations. SYMP 1. Development and Characterization of Multifunctional Materials. SYMP 2. Mechanics and Behavior of Active Materials. SYMP 3. Modeling, Simulation and Control of Adaptive Systems. SYMP 4. Integrated System Design and Implementation. SYMP 5. Structural Health Monitoring. SYMP 6. Bioinspired Smart Materials and Systems. SYMP 7. Energy Harvesting. Authors of selected papers in the materials areas (symposia 1, 2, and 6) as well as energy harvesting (symposium 7) were invited to write a full journal article on their presentation topic for publication in this special issue of Smart

  17. Methylmalonic acid blood test

    MedlinePlus

    ... acid is a substance produced when proteins, called amino acids, in the body break down. The health care ... Cederbaum S, Berry GT. Inborn errors of carbohydrate, ammonia, amino acid, and organic acid metabolism. In: Gleason CA, Devaskar ...

  18. Folic acid - test

    MedlinePlus

    Folic acid is a type of B vitamin. This article discusses the test to measure the amount of folic acid in the blood. ... that may interfere with test results, including folic acid supplements. Drugs that can decrease folic acid measurements ...

  19. Uric acid urine test

    MedlinePlus

    The uric acid urine test measures the level of uric acid in urine. Uric acid level can also be checked using a blood ... help determine the cause of a high uric acid level in the blood. It may also be ...

  20. Methylmalonic acid blood test

    MedlinePlus

    The methylmalonic acid blood test measures the amount of methylmalonic acid in the blood. ... Methylmalonic acid is a substance produced when proteins, called amino acids, in the body break down. The health care ...

  1. Folic Acid and Pregnancy

    MedlinePlus

    ... 5 Things to Know About Zika & Pregnancy Folic Acid and Pregnancy KidsHealth > For Parents > Folic Acid and ... before conception and during early pregnancy . About Folic Acid Folic acid, sometimes called folate, is a B ...

  2. Understanding Acid Rain

    ERIC Educational Resources Information Center

    Damonte, Kathleen

    2004-01-01

    The term acid rain describes rain, snow, or fog that is more acidic than normal precipitation. To understand what acid rain is, it is first necessary to know what an acid is. Acids can be defined as substances that produce hydrogen ions (H+), when dissolved in water. Scientists indicate how acidic a substance is by a set of numbers called the pH…

  3. New bioactive fatty acids

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Many oxygenated fatty acids are bioactive compounds. Nocardia cholesterolicum and Flavobacterium DS5 convert oleic acid to 10 hydroxy stearic acid and linoleic acid to 10-hydroxy-12(Z)-octadecanoic acid. Pseudomonas aeruginosa PR3 converts oleic acid to the new compounds, 7,10-dihydroxy-8(E)-octad...

  4. New Bioactive Fatty Acids

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Many oxygenated fatty acids are bioactive compounds. Nocardia cholesterolicum and Flavobacterium DS5 convert oleic acid to 10 hydroxy stearic acid and linoleic acid to 10-hydroxy-12(Z)-octadecanoic acid. Pseudomonas aeruginosa PR3 converts oleic acid to new compounds, 7,10-dihydroxy-8(E)-octadecen...

  5. Epidemiological, clinical and biochemical characterization of the p.(Ala359Asp) SMPD1 variant causing Niemann-Pick disease type B.

    PubMed

    Acuña, Mariana; Martínez, Pablo; Moraga, Carol; He, Xingxuan; Moraga, Mauricio; Hunter, Bessie; Nuernberg, Peter; Gutiérrez, Rodrigo A; González, Mauricio; Schuchman, Edward H; Santos, José Luis; Miquel, Juan Francisco; Mabe, Paulina; Zanlungo, Silvana

    2016-02-01

    Niemann-Pick disease type B (NPDB) is a rare, inherited lysosomal storage disorder that occurs due to variants in the sphingomyelin phosphodiesterase 1 (SMPD1) gene and the resultant deficiency of acid sphingomyelinase (ASM) activity. While numerous variants causing NPDB have been described, only a small number have been studied in any detail. Herein, we describe the frequency of the p.(Ala359Asp) variant in the healthy Chilean population, and determine the haplotype background of homozygous patients to establish if this variant originated from a common founder. Genomic DNA samples from 1691 healthy individuals were analyzed for the p.(Ala359Asp) variant. The frequency of p.(Ala359Asp) was found to be 1/105.7, predicting a disease incidence of 1/44 960 in Chile, higher than the incidence estimated by the number of confirmed NPDB cases. We also describe the clinical characteristics of 13 patients homozygous for p.(Ala359Asp) and all of them had moderate to severe NPDB disease. In addition, a conserved haplotype and shared 280 Kb region around the SMPD1 gene was observed in the patients analyzed, indicating that the variant originated from a common ancestor. The haplotype frequency and mitochondrial DNA analysis suggest an Amerindian origin for the variant. To assess the effect of the p.(Ala359Asp) variant, we transfected cells with the ASM-p.(Ala359Asp) cDNA and the activity was only 4.2% compared with the wild-type cDNA, definitively demonstrating the causative effect of the variant on ASM function. Information on common variants such as p.(Ala359Asp) is essential to guide the successful implementation for future therapies and benefit to patients. PMID:25920558

  6. ASM News Volume 71 Number 9, 2005

    SciTech Connect

    Tamar Barkay and Barth F. Smets

    2005-01-01

    Genetic exchanges among prokaryotes, formerly considered only a marginal phenomenon, increasingly are being viewed as profoundly affecting evolution. Indeed, some researchers argue for utterly revamping our concept of microbial speciation and phylogeny by replacing the traditional ''tree'' with a newer ''net'' to account for these horizontal transfers of genes. This conceptual ferment is occurring while molecular biologists reveal how horizontal gene transfers occur even as microbes protect the integrity of their genomes. Other studies reveal the number and diversity and abundance of genetic elements that mediate horizontal gene transfers (HGTs) or facilitate genome rearrangements, deletions, and insertions. Taken together, this information suggests that microbial communities collectively possess a dynamic gene pool, where novel genetic combinations act as a driving force in genomic innovation, compensating individual microbial species for their inability to reproduce sexually. These microbial genomic dynamics can present both environmental threats and promise to humans. One major threat, for example, comes from the spread of antibiotic resistance and virulence genes among pathogenic microbes. Another less-documented issue involves transgenic plants and animals, whose uses are being restricted because of concerns that genes may be transferred to untargeted organisms where they might cause harm. A possible benefit from HGT comes from its potential to enhance the functional diversity of microbial communities and to improve their performance in changing or extreme environments. Such changes might be exploited, for example, as part of efforts to manage environmental pollution and might be achieved by spreading genes into resident microbes to confer specific biochemical activities.

  7. ASME Code Calculations for the CC Cryostat

    SciTech Connect

    Luther, R.D.; /Fermilab

    1987-11-04

    This engineering note contains the ASHE Code calculations for the CC Cryostat prepared by the manufacturer, Richmond-Lox Equipment Company. Most of these were taken from calculations initially prepared by Fermilab personne1and pub1ished in Eng. Note 68.

  8. ASM Student Technology and Career Night

    NASA Technical Reports Server (NTRS)

    Hamilton, Jeff

    2005-01-01

    This viewgraph presentation presents a general overview of Marshall Space Flight Center (MSFC) for students who are perspective MSFC employees. The presentation includes an organizational chart and a summary of MSFC activities, as well as photographs and descriptions of some of the center's test facilities.

  9. 1994 ASME/IEEE Joint Railroad Conference

    SciTech Connect

    Hawthorne, K.L.; Hill, J.R.

    1994-12-31

    This conference proceedings contains 21 papers. Topics discussed include railroad transportation, signal systems, computer systems, control systems, power supply system, vehicle suspensions, vehicle wheels, traction, failure (mechanical) and sensors. Some papers have been processed separately for inclusion on the data base.

  10. Adaptive and active materials: selected papers from the ASME 2012 Conference on Smart Materials, Adaptive Structures and Intelligent Systems (SMASIS 12) (Stone Mountain, GA, USA, 19-21 September 2012)

    NASA Astrophysics Data System (ADS)

    Seelecke, Stefan; Erturk, Alper; Ounaies, Zoubeida; Naguib, Hani; Huber, John; Turner, Travis; Anderson, Iain; Philen, Michael; Baba Sundaresan, Vishnu

    2013-09-01

    The fifth annual meeting of the ASME/AIAA Smart Materials, Adaptive Structures and Intelligent Systems Conference (SMASIS) was held in beautiful Stone Mountain near Atlanta, GA. It is the conference's objective to provide an up-to-date overview of research trends in the entire field of smart materials systems. This was reflected in keynote speeches by Professor Eduard Arzt (Institute of New Materials and Saarland University, Saarbrücken, Germany) on 'Micro-patterned artificial 'Gecko' surfaces: a path to switchable adhesive function', by Professor Ray H Baughman (The Alan G MacDiarmid NanoTech Institute, University of Texas at Dallas) on 'The diverse and growing family of carbon nanotube and related artificial muscles', and by Professor Richard James (University of Minnesota) on 'The direct conversion of heat to electricity using multiferroic materials with phase transformations'. SMASIS 2012 was divided into eight symposia which span basic research, applied technological design and development, and industrial and governmental integrated system and application demonstrations. • SYMP 1. Development and characterization of multifunctional materials. • SYMP 2. Mechanics and behavior of active materials. • SYMP 3. Modeling, simulation and control of adaptive systems. • SYMP 4. Integrated system design and implementation. • SYMP 5. Structural health monitoring/NDE. • SYMP 6. Bio-inspired materials and systems. • SYMP 7. Energy harvesting. • SYMP 8. Structural and materials logic. This year we were particularly excited to introduce a new symposium on energy harvesting, which has quickly matured from a special track in previous years to an independent symposium for the first time. The subject cuts across fields by studying different materials, ranging from piezoelectrics to electroactive polymers, as well as by emphasizing different energy sources from wind to waves and ambient vibrations. Modeling, experimental studies, and technology applications all

  11. Bioactive Fatty Acids

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Oxygenated fatty acids are useful as specialty chemicals, plasticizers, and biomedicals. Microbial enzymes convert fatty acids to mono-, di-, and trihydroxy fatty acid products. Among them, Bacillus megaterium ALA2 converted n-6 and n-3 PUFAs to many new oxygenated fatty acids. Linoleic acid was ...

  12. Uric acid test (image)

    MedlinePlus

    Uric acid urine test is performed to check for the amount of uric acid in urine. Urine is collected over a 24 ... testing. The most common reason for measuring uric acid levels is in the diagnosis or treatment of ...

  13. Amino Acid Metabolism Disorders

    MedlinePlus

    ... defects & other health conditions > Amino acid metabolism disorders Amino acid metabolism disorders E-mail to a friend Please ... baby’s newborn screening may include testing for certain amino acid metabolism disorders. These are rare health conditions that ...

  14. Plasma amino acids

    MedlinePlus

    Amino acids blood test ... types of methods used to determine the individual amino acid levels in the blood. ... test is done to measure the level of amino acids in the blood. An increased level of a ...

  15. Stomach acid test

    MedlinePlus

    Gastric acid secretion test ... of the cells in the stomach to release acid. The stomach contents are then removed and analyzed. ... 3.5). These numbers are converted to actual acid production in units of milliequivalents per hour in ...

  16. Azelaic Acid Topical

    MedlinePlus

    Azelaic acid gel is used to clear the bumps, lesions, and swelling caused by rosacea (a skin disease that ... redness, flushing, and pimples on the face). Azelaic acid cream is used to treat acne. Azelaic acid ...

  17. Facts about Folic Acid

    MedlinePlus

    ... Information For... Media Policy Makers Facts About Folic Acid Language: English Español (Spanish) Recommend on Facebook Tweet ... of the baby's brain and spine. About folic acid Folic acid is a B vitamin. Our bodies ...

  18. Acid Lipase Disease

    MedlinePlus

    ... Awards Enhancing Diversity Find People About NINDS NINDS Acid Lipase Disease Information Page Synonym(s): Cholesterol Ester Storage ... Trials Related NINDS Publications and Information What is Acid Lipase Disease ? Acid lipase disease or deficiency occurs ...

  19. Folic acid - test

    MedlinePlus

    ... folic acid measurements include: Alcohol Aminosalicylic acid Birth control pills Estrogens Tetracyclines Ampicillin Chloramphenicol Erythromycin Methotrexate Penicillin Aminopterin Phenobarbital Phenytoin Drugs to treat malaria

  20. Oxalic acid poisoning

    MedlinePlus

    Symptoms of oxalic acid poisoning include: Abdominal pain Burns and blisters where the acid contacted the skin Collapse Convulsions Mouth pain Shock Throat pain Tremors (unintentional trembling) Vomiting

  1. Acid distribution in phosphoric acid fuel cells

    SciTech Connect

    Okae, I.; Seya, A.; Umemoto, M.

    1996-12-31

    Electrolyte acid distribution among each component of a cell is determined by capillary force when the cell is not in operation, but the distribution under the current load conditions had not been clear so far. Since the loss of electrolyte acid during operation is inevitable, it is necessary to store enough amount of acid in every cell. But it must be under the level of which the acid disturbs the diffusion of reactive gases. Accordingly to know the actual acid distribution during operation in a cell is very important. In this report, we carried out experiments to clarify the distribution using small single cells.

  2. Acid tolerance in amphibians

    SciTech Connect

    Pierce, B.A.

    1985-04-01

    Studies of amphibian acid tolerance provide information about the potential effects of acid deposition on amphibian communities. Amphibians as a group appear to be relatively acid tolerant, with many species suffering increased mortality only below pH 4. However, amphibians exhibit much intraspecific variation in acid tolerance, and some species are sensitive to even low levels of acidity. Furthermore, nonlethal effects, including depression of growth rates and increases in developmental abnormalities, can occur at higher pH.

  3. The hyaluronic acid inhibitor 4-methylumbelliferone is an NSMase2 activator-role of Ceramide in MU anti-tumor activity.

    PubMed

    Qin, Jingdong; Kilkus, John; Dawson, Glyn

    2016-02-01

    Increased synthesis of hyaluronic acid (HA) is often associated with increased metastatic potential and invasivity of tumor cells. 4-Methylumbelliferone (MU) is an inhibitor of HA synthesis, and has been studied as a potential anti-tumor drug to inhibit the growth of primary tumors and distant metastasis of tumor cells. Although several studies reported that the anticancer effects of MU are mediated by inhibition of HA signaling, the mechanism still needs to be clarified. In a previous study we demonstrated the regulation of HA synthesis by ceramide, and now show how MU activated neutral sphingomyelinase2 (NSMase2) generates ceramides and mediates MU induced inhibition of HA synthesis, cell migration and invasion, and apoptosis of tumor cells. Using a HA enriched mouse oligodendroglioma cell line G26-24 we found that MU elevated the activity of NSMase2 and increased ceramide levels, which in turn increased phosphatase PP2A activity. Further, the activated PP2A reduced phosphorylation of Akt, decreased activities of HA synthase2 (HAS2) and calpains, and inhibited both the synthesis of HA, and the migration and invasion of G26-24 tumor cells. In addition, MU mediated ceramide stimulated activation of p53 and caspase-3, reduced SIRT1 expression and decreased G26-24 viability. The mechanism of the MU anticancer therefore initially involves NSMase2/ceramide/PP2A/AKT/HAS2/caspase-3/p53/SIRT1 and the calpain signaling pathway, suggesting that ceramides play a key role in the ability of a tumor to become aggressively metastatic and grow. PMID:26548718

  4. Toxicity of adipic acid.

    PubMed

    Kennedy, Gerald L

    2002-05-01

    Adipic acid has very low acute toxicity in rats with an LD50 > 5000 mg/kg. Adipic acid produced mild to no skin irritation on intact guinea pig skin as a 50% concentration in propylene glycol; it was not a skin sensitizer. Adipic acid caused mild conjunctival irritation in washed rabbit eyes; in unwashed rabbit eyes, there was mild conjunctival irritation, minimal iritis, but no corneal effects. Adipic acid dust may irritate the mucous membranes of the lungs and nose. In a 2-year feeding study, rats fed adipic acid at concentrations up to 5% in the diet exhibited only weight loss. Adipic acid is not genetically active in a wide variety of assay systems. Adipic acid caused no developmental toxicity in mice, rats, rabbits, or hamsters when administered orally. Adipic acid is partially metabolized in humans; the balance is eliminated unchanged in the urine. Adipic acid is slightly to moderately toxic to fish, daphnia, and algae in acute tests. PMID:12024802

  5. Acid Thunder: Acid Rain and Ancient Mesoamerica

    ERIC Educational Resources Information Center

    Kahl, Jonathan D. W.; Berg, Craig A.

    2006-01-01

    Much of Mesoamerica's rich cultural heritage is slowly eroding because of acid rain. Just as water dissolves an Alka-Seltzer tablet, acid rain erodes the limestone surfaces of Mexican archaeological sites at a rate of about one-half millimeter per century (Bravo et al. 2003). A half-millimeter may not seem like much, but at this pace, a few…

  6. Quantity of acid in acid fog

    SciTech Connect

    Deal, W.J.

    1983-07-01

    This communication notes the actual magnitude of the acidity in acidic fog particles and suggests a possible line of inquiry into the health effects of such fog so that it can be determined whether a typical fog is detrimental or beneficial relative to dry air.

  7. [Amino acids in saliva].

    PubMed

    Klinger, G; Gruhn, K

    1984-01-01

    Total amino acids in saliva and free and peptide-bound amino acids from 21 saliva samples were determined. The contents of amino acids was 25 mmol/1; total nitrogen content was 78-80 mmol/1. Amino acids consist of Prolin in 25%. Some patients were examined before and after application of the depot estrogen ethinyl estradiosulfonat, which stimulates the assimilation of protein. After application, amino acids increased and the authors found a shift between the single amino acids. Estrogen medication induced an increase in proteins with the character of collagens. Clinical effects are discussed. (author's modified) PMID:6240853

  8. Hydrochloric acid poisoning

    MedlinePlus

    Hydrochloric acid is a clear, poisonous liquid. It is highly corrosive, which means it immediately causes severe damage, ... discusses poisoning due to swallowing or breathing in hydrochloric acid. This article is for information only. Do NOT ...

  9. Omega-3 Fatty Acids

    MedlinePlus

    Omega-3 fatty acids are used together with lifestyle changes (diet, weight-loss, exercise) to reduce the amount ... the blood in people with very high triglycerides. Omega-3 fatty acids are in a class of medications ...

  10. Omega-6 Fatty Acids

    MedlinePlus

    Omega-6 fatty acids are types of fats. Some types are found in vegetable oils, including corn, evening primrose seed, safflower, and soybean oils. Other types of omega-6 fatty acids are found in black currant seed, borage seed, ...

  11. Zoledronic Acid Injection

    MedlinePlus

    Zoledronic acid (Reclast) is used to prevent or treat osteoporosis (condition in which the bones become thin and weak ... of life,' end of regular menstrual periods). Zoledronic acid (Reclast) is also used to treat osteoporosis in ...

  12. Aminolevulinic Acid Topical

    MedlinePlus

    Aminolevulinic acid is used in combination with photodynamic therapy (PDT; special blue light) to treat actinic keratoses (small crusty ... skin cancer) of the face or scalp. Aminolevulinic acid is in a class of medications called photosensitizing ...

  13. Acid-fast stain

    MedlinePlus

    The acid-fast stain is a laboratory test that determines if a sample of tissue, blood, or other body ... dye. The slide is then washed with an acid solution and a different stain is applied. Bacteria ...

  14. Uric acid - blood

    MedlinePlus

    Uric acid is a chemical created when the body breaks down substances called purines. Purines are found in some ... dried beans and peas, and beer. Most uric acid dissolves in blood and travels to the kidneys. ...

  15. Hydrochloric acid poisoning

    MedlinePlus

    Hydrochloric acid is a clear, poisonous liquid. It is highly corrosive, which means it immediately causes severe damage, such ... poisoning due to swallowing or breathing in hydrochloric acid. This article is for information only. Do NOT ...

  16. Omega-6 Fatty Acids

    MedlinePlus

    ... types of fats. Some types are found in vegetable oils, including corn, evening primrose seed, safflower, and soybean ... from studying specific omega-6 fatty acids or plant oils containing omega-6 fatty acids. See the separate ...

  17. Uric Acid Test

    MedlinePlus

    ... limited. Home Visit Global Sites Search Help? Uric Acid Share this page: Was this page helpful? Also known as: Serum Urate; UA Formal name: Uric Acid Related tests: Synovial Fluid Analysis , Kidney Stone Analysis , ...

  18. Acid-fast stain

    MedlinePlus

    ... this page: //medlineplus.gov/ency/article/003766.htm Acid-fast stain To use the sharing features on this page, please enable JavaScript. The acid-fast stain is a laboratory test that determines ...

  19. Aminocaproic Acid Injection

    MedlinePlus

    Aminocaproic acid injection is used to control bleeding that occurs when blood clots are broken down too quickly. This ... the baby is ready to be born). Aminocaproic acid injection is also used to control bleeding in ...

  20. Deoxycholic Acid Injection

    MedlinePlus

    Deoxycholic acid injection is used to improve the appearance and profile of moderate to severe submental fat ('double chin'; fatty tissue located under the chin). Deoxycholic acid injection is in a class of medications called ...

  1. Methylmalonic Acid Test

    MedlinePlus

    ... limited. Home Visit Global Sites Search Help? Methylmalonic Acid Share this page: Was this page helpful? Also known as: MMA Formal name: Methylmalonic Acid Related tests: Vitamin B12 and Folate , Homocysteine , Intrinsic ...

  2. Fatty acid analogs

    DOEpatents

    Elmaleh, David R.; Livni, Eli

    1985-01-01

    In one aspect, a radioactively labeled analog of a fatty acid which is capable of being taken up by mammalian tissue and which exhibits an in vivo beta-oxidation rate below that with a corresponding radioactively labeled fatty acid.

  3. Boric acid poisoning

    MedlinePlus

    ... boric acid poisoning usually occurs when someone swallows powdered roach-killing products that contain the chemical. Chronic ... vein (IV) Medicines to treat symptoms Note: Activated charcoal does not effectively treat (absorb) boric acid. For ...

  4. Lactic acid test

    MedlinePlus

    ... this page: //medlineplus.gov/ency/article/003507.htm Lactic acid test To use the sharing features on this page, please enable JavaScript. Lactic acid is mainly produced in muscle cells and red ...

  5. PRODUCTION OF TRIFLUOROACETIC ACID

    DOEpatents

    Haworth, W.N.; Stacey, M.

    1949-07-19

    A method is given for the production of improved yields of trifluoroacetic acid. The compound is prepared by oxidizing m-aminobenzotrifluoride with an alkali metal or alkaline earth metal permanganate at a temperature in the range of 80 deg C to 100 deg C while dissolved ln a mixture of water with glacial acetic acid and/or trifluoroacetic acid. Preferably a mixture of water and trifluoroacetic acid ls used as the solvent.

  6. Plant fatty acid hydroxylases

    DOEpatents

    Somerville, Chris; Broun, Pierre; van de Loo, Frank

    2001-01-01

    This invention relates to plant fatty acyl hydroxylases. Methods to use conserved amino acid or nucleotide sequences to obtain plant fatty acyl hydroxylases are described. Also described is the use of cDNA clones encoding a plant hydroxylase to produce a family of hydroxylated fatty acids in transgenic plants. In addition, the use of genes encoding fatty acid hydroxylases or desaturases to alter the level of lipid fatty acid unsaturation in transgenic plants is described.

  7. Quantity of acid in acid fog

    SciTech Connect

    Deal, W.J.

    1983-07-01

    The chemical composition of fog particles has become of considerable interest, because of both the possibility of interpreting atmospheric- chemistry processes in fog particles in terms of the principles of aqueous chemistry and the potential health effects of species present in fog particles. The acidity of fog particles has received wide attention. This communication noted the actual magnitude of the excess acidity in acidic fog particles and suggested a possible line of inquiry into the health effects of such fog so that it can be determined whether a typical fog is detrimental or beneficial relative to dry air. (DP)

  8. The Acid Rain Reader.

    ERIC Educational Resources Information Center

    Stubbs, Harriett S.; And Others

    A topic which is often not sufficiently dealt with in elementary school textbooks is acid rain. This student text is designed to supplement classroom materials on the topic. Discussed are: (1) "Rain"; (2) "Water Cycle"; (3) "Fossil Fuels"; (4) "Air Pollution"; (5) "Superstacks"; (6) "Acid/Neutral/Bases"; (7) "pH Scale"; (8) "Acid Rain"; (9)…

  9. What Is Acid Rain?

    ERIC Educational Resources Information Center

    Likens, Gene E.

    2004-01-01

    Acid rain is the collective term for any type of acidified precipitation: rain, snow, sleet, and hail, as well as the presence of acidifying gases, particles, cloud water, and fog in the atmosphere. The increased acidity, primarily from sulfuric and nitric acids, is generated as a by-product of the combustion of fossil fuels such as coal and oil.…

  10. Acid Rain Study Guide.

    ERIC Educational Resources Information Center

    Hunger, Carolyn; And Others

    Acid rain is a complex, worldwide environmental problem. This study guide is intended to aid teachers of grades 4-12 to help their students understand what acid rain is, why it is a problem, and what possible solutions exist. The document contains specific sections on: (1) the various terms used in conjunction with acid rain (such as acid…

  11. [alpha]-Oxocarboxylic Acids

    ERIC Educational Resources Information Center

    Kerber, Robert C.; Fernando, Marian S.

    2010-01-01

    Several [alpha]-oxocarboxylic acids play key roles in metabolism in plants and animals. However, there are inconsistencies between the structures as commonly portrayed and the reported acid ionization constants, which result because the acids are predominantly hydrated in aqueous solution; that is, the predominant form is RC(OH)[subscript 2]COOH…

  12. Cleavage of nucleic acids

    DOEpatents

    Prudent, James R.; Hall, Jeff G.; Lyamichev, Victor L.; Brow, Mary Ann D.; Dahlberg, James E.

    2007-12-11

    The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The structure-specific nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof.

  13. Amino acid analysis

    NASA Technical Reports Server (NTRS)

    Winitz, M.; Graff, J. (Inventor)

    1974-01-01

    The process and apparatus for qualitative and quantitative analysis of the amino acid content of a biological sample are presented. The sample is deposited on a cation exchange resin and then is washed with suitable solvents. The amino acids and various cations and organic material with a basic function remain on the resin. The resin is eluted with an acid eluant, and the eluate containing the amino acids is transferred to a reaction vessel where the eluant is removed. Final analysis of the purified acylated amino acid esters is accomplished by gas-liquid chromatographic techniques.

  14. Editorial: Acid precipitation

    SciTech Connect

    1995-09-01

    This editorial focuses on acid rain and the history of public and governmental response to acid rain. Comments on a book by Gwineth Howell `Acid Rain and Acid Waters` are included. The editor feels that Howells has provide a service to the environmental scientific community, with a textbook useful to a range of people, as well as a call for decision makers to learn from the acid rain issue and use it as a model for more sweeping global environmental issues. A balance is needed among several parameters such as level of evidence, probability that the evidence will lead to a specific direction and the cost to the global community. 1 tab.

  15. Cleavage of nucleic acids

    SciTech Connect

    Prudent, James R.; Hall, Jeff G.; Lyamichev, Victor I.; Brow; Mary Ann D.; Dahlberg, James E.

    2010-11-09

    The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The structure-specific nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof.

  16. Cleavage of nucleic acids

    DOEpatents

    Prudent, James R.; Hall, Jeff G.; Lyamichev, Victor I.; Brow, Mary Ann D.; Dahlberg, James E.

    2000-01-01

    The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The structure-specific nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof.

  17. Nucleic acid detection assays

    DOEpatents

    Prudent, James R.; Hall, Jeff G.; Lyamichev, Victor I.; Brow, Mary Ann; Dahlberg, James E.

    2005-04-05

    The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The structure-specific nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof.

  18. Nucleic acid detection compositions

    DOEpatents

    Prudent, James R.; Hall, Jeff G.; Lyamichev, Victor I.; Brow, Mary Ann; Dahlberg, James L.

    2008-08-05

    The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The structure-specific nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof.

  19. Unusual Mn(III/IV)4 Cubane and Mn(III)16M4 (M = Ca, Sr) Looplike Clusters from the Use of Dimethylarsinic Acid.

    PubMed

    Chakov, Nicole E; Thuijs, Annaliese E; Wernsdorfer, Wolfgang; Rheingold, Arnold L; Abboud, Khalil A; Christou, George

    2016-09-01

    Three complexes are reported from the initial use of dimethylarsinic acid (Me2AsO2H) in Mn(III/IV) cluster chemistry, [Mn4O4(O2AsMe2)6] (3; 2Mn(III), 2Mn(IV)), and [Mn16X4O8(O2CPh)16(Me2AsO2)24] (X = Ca(2+) (4) or Sr(2+) (5); 16Mn(III)). They were obtained from reactions with [Mn12O12(O2CR)16(H2O)4] (R = Me, Ph) either without (3) or with (4 and 5) the addition of X(2+) salts. Complex 3 contains a [Mn4O4](6+) cubane, whereas isostructural 4 and 5 contain a planar loop structure comprising four Mn4 asymmetric "butterfly" units linked by alternating anti,anti μ-O2AsMe2 and {X2(O2AsMe2)(O2CPh)2} units. Variable-temperature magnetic susceptibility (χM) data were collected on dried microcrystalline samples of 3-5 in the 5.0-300 K range in a 0.1 T (1000 G) direct-current (dc) magnetic field. Data for 3 were fit to the appropriate Van Vleck equation (using the [Formula: see text] = -2JŜi·Ŝj convention) for a cubane of virtual C2v symmetry, giving J33 = 0.0(1) cm(-1), J34 = -3.4(4) cm(-1), J44 = -9.8(2) cm(-1), and g = 1.99(1), where the Jij subscripts refer to the oxidation states of the interacting Mn atoms. The ground state thus consists of two coupled Mn(IV) and two essentially noninteracting Mn(III). For 4 and 5, low-lying excited states from the high nuclearity and weak couplings prevent fits of dc magnetization data, but in-phase alternating current susceptibility χ'MT data down to 1.8 K indicate them to possess S = 4 ground states, if considered single Mn16 units. If instead they are treated as tetramers of weakly coupled Mn4 units, then each of the latter has an S = 2 ground state. Complexes 4 and 5 also exhibit very weak out-of-phase χ″M signals characteristic of slow relaxation, and magnetization versus dc field scans on a single crystal of 4·15MeCN at T ≥ 0.04 K showed hysteresis loops but with unusual features suggesting the magnetization relaxation barrier consists of more than one contribution. PMID:27504743

  20. Acidic Ionic Liquids.

    PubMed

    Amarasekara, Ananda S

    2016-05-25

    Ionic liquid with acidic properties is an important branch in the wide ionic liquid field and the aim of this article is to cover all aspects of these acidic ionic liquids, especially focusing on the developments in the last four years. The structural diversity and synthesis of acidic ionic liquids are discussed in the introduction sections of this review. In addition, an unambiguous classification system for various types of acidic ionic liquids is presented in the introduction. The physical properties including acidity, thermo-physical properties, ionic conductivity, spectroscopy, and computational studies on acidic ionic liquids are covered in the next sections. The final section provides a comprehensive review on applications of acidic ionic liquids in a wide array of fields including catalysis, CO2 fixation, ionogel, electrolyte, fuel-cell, membrane, biomass processing, biodiesel synthesis, desulfurization of gasoline/diesel, metal processing, and metal electrodeposition. PMID:27175515

  1. Nucleic acid detection kits

    DOEpatents

    Hall, Jeff G.; Lyamichev, Victor I.; Mast, Andrea L.; Brow, Mary Ann; Kwiatkowski, Robert W.; Vavra, Stephanie H.

    2005-03-29

    The present invention relates to means for the detection and characterization of nucleic acid sequences, as well as variations in nucleic acid sequences. The present invention also relates to methods for forming a nucleic acid cleavage structure on a target sequence and cleaving the nucleic acid cleavage structure in a site-specific manner. The structure-specific nuclease activity of a variety of enzymes is used to cleave the target-dependent cleavage structure, thereby indicating the presence of specific nucleic acid sequences or specific variations thereof. The present invention further relates to methods and devices for the separation of nucleic acid molecules based on charge. The present invention also provides methods for the detection of non-target cleavage products via the formation of a complete and activated protein binding region. The invention further provides sensitive and specific methods for the detection of nucleic acid from various viruses in a sample.

  2. Demospongic Acids Revisited

    PubMed Central

    Kornprobst, Jean-Michel; Barnathan, Gilles

    2010-01-01

    The well-known fatty acids with a Δ5,9 unsaturation system were designated for a long period as demospongic acids, taking into account that they originally occurred in marine Demospongia sponges. However, such acids have also been observed in various marine sources with a large range of chain-lengths (C16–C32) and from some terrestrial plants with short acyl chains (C18–C19). Finally, the Δ5,9 fatty acids appear to be a particular type of non-methylene-interrupted fatty acids (NMA FAs). This article reviews the occurrence of these particular fatty acids in marine and terrestrial organisms and shows the biosynthetic connections between Δ5,9 fatty acids and other NMI FAs. PMID:21116406

  3. Process for the preparation of lactic acid and glyceric acid

    DOEpatents

    Jackson, James E [Haslett, MI; Miller, Dennis J [Okemos, MI; Marincean, Simona [Dewitt, MI

    2008-12-02

    Hexose and pentose monosaccharides are degraded to lactic acid and glyceric acid in an aqueous solution in the presence of an excess of a strongly anionic exchange resin, such as AMBERLITE IRN78 and AMBERLITE IRA400. The glyceric acid and lactic acid can be separated from the aqueous solution. Lactic acid and glyceric acid are staple articles of commerce.

  4. Microorganisms for producing organic acids

    SciTech Connect

    Pfleger, Brian Frederick; Begemann, Matthew Brett

    2014-09-30

    Organic acid-producing microorganisms and methods of using same. The organic acid-producing microorganisms comprise modifications that reduce or ablate AcsA activity or AcsA homolog activity. The modifications increase tolerance of the microorganisms to such organic acids as 3-hydroxypropionic acid, acrylic acid, propionic acid, lactic acid, and others. Further modifications to the microorganisms increase production of such organic acids as 3-hydroxypropionic acid, lactate, and others. Methods of producing such organic acids as 3-hydroxypropionic acid, lactate, and others with the modified microorganisms are provided. Methods of using acsA or homologs thereof as counter-selectable markers are also provided.

  5. Acid-Base Homeostasis

    PubMed Central

    Nakhoul, Nazih; Hering-Smith, Kathleen S.

    2015-01-01

    Acid-base homeostasis and pH regulation are critical for both normal physiology and cell metabolism and function. The importance of this regulation is evidenced by a variety of physiologic derangements that occur when plasma pH is either high or low. The kidneys have the predominant role in regulating the systemic bicarbonate concentration and hence, the metabolic component of acid-base balance. This function of the kidneys has two components: reabsorption of virtually all of the filtered HCO3− and production of new bicarbonate to replace that consumed by normal or pathologic acids. This production or generation of new HCO3− is done by net acid excretion. Under normal conditions, approximately one-third to one-half of net acid excretion by the kidneys is in the form of titratable acid. The other one-half to two-thirds is the excretion of ammonium. The capacity to excrete ammonium under conditions of acid loads is quantitatively much greater than the capacity to increase titratable acid. Multiple, often redundant pathways and processes exist to regulate these renal functions. Derangements in acid-base homeostasis, however, are common in clinical medicine and can often be related to the systems involved in acid-base transport in the kidneys. PMID:26597304

  6. Citric Acid Alternative to Nitric Acid Passivation

    NASA Technical Reports Server (NTRS)

    Lewis, Pattie L. (Compiler)

    2013-01-01

    The Ground Systems Development and Operations GSDO) Program at NASA John F. Kennedy Space Center (KSC) has the primary objective of modernizing and transforming the launch and range complex at KSC to benefit current and future NASA programs along with other emerging users. Described as the launch support and infrastructure modernization program in the NASA Authorization Act of 2010, the GSDO Program will develop and implement shared infrastructure and process improvements to provide more flexible, affordable, and responsive capabilities to a multi-user community. In support of the GSDO Program, the purpose of this project is to demonstratevalidate citric acid as a passivation agent for stainless steel. Successful completion of this project will result in citric acid being qualified for use as an environmentally preferable alternative to nitric acid for passivation of stainless steel alloys in NASA and DoD applications.

  7. USGS Tracks Acid Rain

    USGS Publications Warehouse

    Gordon, John D.; Nilles, Mark A.; Schroder, LeRoy J.

    1995-01-01

    The U.S. Geological Survey (USGS) has been actively studying acid rain for the past 15 years. When scientists learned that acid rain could harm fish, fear of damage to our natural environment from acid rain concerned the American public. Research by USGS scientists and other groups began to show that the processes resulting in acid rain are very complex. Scientists were puzzled by the fact that in some cases it was difficult to demonstrate that the pollution from automobiles and factories was causing streams or lakes to become more acidic. Further experiments showed how the natural ability of many soils to neutralize acids would reduce the effects of acid rain in some locations--at least as long as the neutralizing ability lasted (Young, 1991). The USGS has played a key role in establishing and maintaining the only nationwide network of acid rain monitoring stations. This program is called the National Atmospheric Deposition Program/National Trends Network (NADP/NTN). Each week, at approximately 220 NADP/NTN sites across the country, rain and snow samples are collected for analysis. NADP/NTN site in Montana. The USGS supports about 72 of these sites. The information gained from monitoring the chemistry of our nation's rain and snow is important for testing the results of pollution control laws on acid rain.

  8. Recovery of organic acids

    DOEpatents

    Verser, Dan W.; Eggeman, Timothy J.

    2009-10-13

    A method is disclosed for the recovery of an organic acid from a dilute salt solution in which the cation of the salt forms an insoluble carbonate salt. A tertiary amine and CO.sub.2 are introduced to the solution to form the insoluble carbonate salt and a complex between the acid and an amine. A water immiscible solvent, such as an alcohol, is added to extract the acid/amine complex from the dilute salt solution to a reaction phase. The reaction phase is continuously dried and a product between the acid and the solvent, such as an ester, is formed.

  9. Recovery of organic acids

    SciTech Connect

    Verser, Dan W.; Eggeman, Timothy J.

    2011-11-01

    A method is disclosed for the recovery of an organic acid from a dilute salt solution in which the cation of the salt forms an insoluble carbonate salt. A tertiary amine and CO.sub.2 are introduced to the solution to form the insoluble carbonate salt and a complex between the acid and an amine. A water immiscible solvent, such as an alcohol, is added to extract the acid/amine complex from the dilute salt solution to a reaction phase. The reaction phase is continuously dried and a product between the acid and the solvent, such as an ester, is formed.

  10. THIN-LAYER SEPARATION OF CITRIC ACID CYCLE INTERMEDIATES, LACTIC ACID, AND THE AMINO ACID TAURINE

    EPA Science Inventory

    This paper describes a two-dimensional mixed-layer method for separating citric acid cycle intermediates, lactic acid and the amino acid taurine. The method cleanly separates all citric acid cycle intermediates tested, excepting citric acid and isocitric acid. The solvents are in...

  11. Attenuation by Statins of Membrane Raft-Redox Signaling in Coronary Arterial EndotheliumS⃞

    PubMed Central

    Wei, Yu-Miao; Li, Xiang; Xiong, Jing; Abais, Justine M.; Xia, Min; Boini, Krishna M.; Li, Pin-Lan

    2013-01-01

    Membrane raft (MR)–redox signaling platforms associated with NADPH oxidase are involved in coronary endothelial dysfunction. Here, we studied whether statins interfere with the formation of MR-redox signaling platforms to protect the coronary arterial endothelium from oxidized low-density lipoprotein (OxLDL)–induced injury and from acute hypercholesterolemia. In cultured human coronary arterial endothelial cells, confocal microscopy detected the formation of an MRs clustering when they were exposed to OxLDL, and such MR platform formation was inhibited markedly by statins, including pravastatin and simvastatin. In these MR clusters, NADPH oxidase subunits gp91phox and p47phox were aggregated and were markedly blocked by both statins. In addition, colocalization of acid sphingomyelinase (ASM) and ceramide was induced by OxLDL, which was blocked by statins. Electron spin resonance spectrometry showed that OxLDL-induced superoxide (O2.−) production in the MR fractions was substantially reduced by statins. In coronary artery intima of mice with acute hypercholesterolemia, confocal microscopy revealed a colocalization of gp91phox, p47phox, ASM, or ceramide in MR clusters. Such colocalization was rarely observed in the arteries of normal mice or significantly reduced by pretreatment of hypercholesterolemic mice with statins. Furthermore, O2.− production in situ was 3-fold higher in the coronary arteries from hypercholesterolemic mice than in those from normal mice, and such increase was inhibited by statins. Our results indicate that blockade of MR-redox signaling platform formation in endothelial cell membrane may be another important therapeutic mechanism of statins in preventing endothelial injury and atherosclerosis and may be associated with their direct action on membrane cholesterol structure and function. PMID:23435541

  12. Toxicology of Perfluoroalkyl acids

    EPA Science Inventory

    The Perfluoroalkyl acids(PFAAs) area a family of organic chemicals consisting of a perflurinated carbon backbone (4-12in length) and a acidic functional moiety (Carboxylate or sulfonate). These compounds have excellent surface-tension reducing properties and have numerous industr...

  13. Toxicology of Perfluoroalkyl Acids*

    EPA Science Inventory

    The perfluoroalkyl acids (PFAAs) are a family of organic chemicals consisting of a perfluorinated carbon backbone (4-12 in length) and an acidic functional moiety (carboxylate or sulfonate). These compounds are chemically stable, have excellent surface-tension reducing properties...

  14. Lead-acid cell

    SciTech Connect

    Hradcovsky, R.J.; Kozak, O.R.

    1980-12-09

    A lead-acid storage battery is described that has a lead negative electrode, a lead dioxide positive electrode and a sulfuric acid electrolyte having an organic catalyst dissolved therein which prevents dissolution of the electrodes into lead sulfate whereby in the course of discharge, the lead dioxide is reduced to lead oxide and the lead is oxidized.

  15. Proteins and Amino Acids

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Proteins are the most abundant substances in living organisms and cells. All proteins are constructed from the same twenty amino acids that are linked together by covalent bonds. Shorter chains of two or more amino acids can be linked by covalent bonds to form polypeptides. There are twenty amino...

  16. EFFECTS OF ACID PRECIPITATION

    EPA Science Inventory

    Recent reviews of available data indicate that precipitation in a large region of North America is highly acidic when its pH is compared with the expected pH value of 5.65 for pure rain water in equilibrium with CO2. A growing body of evidence suggests that acid rain is responsib...

  17. Bile acid transporters

    PubMed Central

    Dawson, Paul A.; Lan, Tian; Rao, Anuradha

    2009-01-01

    In liver and intestine, transporters play a critical role in maintaining the enterohepatic circulation and bile acid homeostasis. Over the past two decades, there has been significant progress toward identifying the individual membrane transporters and unraveling their complex regulation. In the liver, bile acids are efficiently transported across the sinusoidal membrane by the Na+ taurocholate cotransporting polypeptide with assistance by members of the organic anion transporting polypeptide family. The bile acids are then secreted in an ATP-dependent fashion across the canalicular membrane by the bile salt export pump. Following their movement with bile into the lumen of the small intestine, bile acids are almost quantitatively reclaimed in the ileum by the apical sodium-dependent bile acid transporter. The bile acids are shuttled across the enterocyte to the basolateral membrane and effluxed into the portal circulation by the recently indentified heteromeric organic solute transporter, OSTα-OSTβ. In addition to the hepatocyte and enterocyte, subgroups of these bile acid transporters are expressed by the biliary, renal, and colonic epithelium where they contribute to maintaining bile acid homeostasis and play important cytoprotective roles. This article will review our current understanding of the physiological role and regulation of these important carriers. PMID:19498215

  18. Fats and fatty acids

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The absolute fat requirement of the human species is the amount of essential fatty acids needed to maintain optimal fatty acid composition of all tissues and normal eicosanoid synthesis. At most, this requirement is no more than about 5% of an adequate energy intake. However, fat accounts for appro...

  19. Analysis of Organic Acids.

    ERIC Educational Resources Information Center

    Griswold, John R.; Rauner, Richard A.

    1990-01-01

    Presented are the procedures and a discussion of the results for an experiment in which students select unknown carboxylic acids, determine their melting points, and investigate their solubility behavior in water and ethanol. A table of selected carboxylic acids is included. (CW)

  20. EXPOSURES TO ACIDIC AEROSOLS

    EPA Science Inventory

    Ambient monitoring of acid aerosol in four U.S. cities and in a rural region of southern Ontario clearly show distinct periods of strong acidity. easurements made in Kingston, TN, and Stuebenville, OH, resulted in 24-hr H+ ion concentrations exceeding 100 nmole/m3 more than 10 ti...

  1. Mutant fatty acid desaturase

    DOEpatents

    Shanklin, John; Cahoon, Edgar B.

    2004-02-03

    The present invention relates to a method for producing mutants of a fatty acid desaturase having a substantially increased activity towards fatty acid substrates with chains containing fewer than 18 carbons relative to an unmutagenized precursor desaturase having an 18 carbon atom chain length substrate specificity. The method involves inducing one or more mutations in the nucleic acid sequence encoding the precursor desaturase, transforming the mutated sequence into an unsaturated fatty acid auxotroph cell such as MH13 E. coli, culturing the cells in the absence of supplemental unsaturated fatty acids, thereby selecting for recipient cells which have received and which express a mutant fatty acid desaturase with an elevated specificity for fatty acid substrates having chain lengths of less than 18 carbon atoms. A variety of mutants having 16 or fewer carbon atom chain length substrate specificities are produced by this method. Mutant desaturases produced by this method can be introduced via expression vectors into prokaryotic and eukaryotic cells and can also be used in the production of transgenic plants which may be used to produce specific fatty acid products.

  2. Omega-3 Fatty Acids

    MedlinePlus

    Omega-3 fatty acids are used together with lifestyle changes (diet, weight-loss, exercise) to reduce the amount of triglycerides (a fat-like ... people with very high triglycerides. Omega-3 fatty acids are in a class of medications called antilipemic ...

  3. Salicylic Acid Topical

    MedlinePlus

    Propa pH® Peel-Off Acne Mask ... pimples and skin blemishes in people who have acne. Topical salicylic acid is also used to treat ... medications called keratolytic agents. Topical salicylic acid treats acne by reducing swelling and redness and unplugging blocked ...

  4. Amino Acid Crossword Puzzle

    ERIC Educational Resources Information Center

    Sims, Paul A.

    2011-01-01

    Learning the 20 standard amino acids is an essential component of an introductory course in biochemistry. Later in the course, the students study metabolism and learn about various catabolic and anabolic pathways involving amino acids. Learning new material or concepts often is easier if one can connect the new material to what one already knows;…

  5. (-)-Oleocanthal rapidly and selectively induces cancer cell death via lysosomal membrane permeabilization

    PubMed Central

    LeGendre, Onica; Breslin, Paul AS; Foster, David A

    2015-01-01

    (-)-Oleocanthal (OC), a phenolic compound present in extra-virgin olive oil (EVOO), has been implicated in the health benefits associated with diets rich in EVOO. We investigated the effect of OC on human cancer cell lines in culture and found that OC induced cell death in all cancer cells examined as rapidly as 30 minutes after treatment in the absence of serum. OC treatment of non-transformed cells suppressed their proliferation but did not cause cell death. OC induced both primary necrotic and apoptotic cell death via induction of lysosomal membrane permeabilization (LMP). We provide evidence that OC promotes LMP by inhibiting acid sphingomyelinase (ASM) activity, which destabilizes the interaction between proteins required for lysosomal membrane stability. The data presented here indicate that cancer cells, which tend to have fragile lysosomal membranes compared to non-cancerous cells, are susceptible to cell death induced by lysosomotropic agents. Therefore, targeting lysosomal membrane stability represents a novel approach for the induction of cancer-specific cell death. PMID:26380379

  6. Trans Fatty Acids

    NASA Astrophysics Data System (ADS)

    Doyle, Ellin

    1997-09-01

    Fats and their various fatty acid components seem to be a perennial concern of nutritionists and persons concerned with healthful diets. Advice on the consumption of saturated, polyunsaturated, monounsaturated, and total fat bombards us from magazines and newspapers. One of the newer players in this field is the group of trans fatty acids found predominantly in partially hydrogenated fats such as margarines and cooking fats. The controversy concerning dietary trans fatty acids was recently addressed in an American Heart Association (AHA) science advisory (1) and in a position paper from the American Society of Clinical Nutrition/American Institute of Nutrition (ASCN/AIN) (2). Both reports emphasize that the best preventive strategy for reducing risk for cardiovascular disease and some types of cancer is a reduction in total and saturated fats in the diet, but a reduction in the intake of trans fatty acids was also recommended. Although the actual health effects of trans fatty acids remain uncertain, experimental evidence indicates that consumption of trans fatty acids adversely affects serum lipid levels. Since elevated levels of serum cholesterol and triacylglycerols are associated with increased risk of cardiovascular disease, it follows that intake of trans fatty acids should be minimized.

  7. Sulfuric Acid on Europa

    NASA Technical Reports Server (NTRS)

    1999-01-01

    Frozen sulfuric acid on Jupiter's moon Europa is depicted in this image produced from data gathered by NASA's Galileo spacecraft. The brightest areas, where the yellow is most intense, represent regions of high frozen sulfuric acid concentration. Sulfuric acid is found in battery acid and in Earth's acid rain.

    This image is based on data gathered by Galileo's near infrared mapping spectrometer.

    Europa's leading hemisphere is toward the bottom right, and there are enhanced concentrations of sulfuric acid in the trailing side of Europa (the upper left side of the image). This is the face of Europa that is struck by sulfur ions coming from Jupiter's innermost moon, Io. The long, narrow features that crisscross Europa also show sulfuric acid that may be from sulfurous material extruded in cracks.

    Galileo, launched in 1989, has been orbiting Jupiter and its moons since December 1995. JPL manages the Galileo mission for NASA's Office of Space Science, Washington DC. JPL is a division of the California Institute of Technology, Pasadena, CA.

  8. Strongly Acidic Auxin Indole-3-Methanesulfonic Acid

    PubMed Central

    Cohen, Jerry D.; Baldi, Bruce G.; Bialek, Krystyna

    1985-01-01

    A radiochemical synthesis is described for [14C]indole-3-methanesulfonic acid (IMS), a strongly acidic auxin analog. Techniques were developed for fractionation and purification of IMS using normal and reverse phase chromatography. In addition, the utility of both Fourier transform infrared spectrometry and fast atom bombardment mass spectrometry for analysis of IMS has been demonstrated. IMS was shown to be an active auxin, stimulating soybean hypocotyl elongation, bean first internode curvature, and ethylene production. IMS uptake by thin sections of soybean hypocotyl was essentially independent of solution pH and, when applied at a 100 micromolar concentration, IMS exhibited a basipetal polarity in its transport in both corn coleoptile and soybean hypocotyl sections. [14C]IMS should, therefore, be a useful compound to study fundamental processes related to the movement of auxins in plant tissues and organelles. PMID:16664007

  9. Understanding acid rain

    SciTech Connect

    Budiansky, S.

    1981-06-01

    The complexities of the phenomenon of acid rain are described. Many factors, including meteorology, geology, chemistry, and biology, all play parts. Varying weather, varying soils, the presence of other pollutants and species differences all act to blur the connections between industrial emissions, acid rain, and environmental damage. Some experts believe that the greatest pH shock to lakes occurs during snow melt and runoff in the spring; others believe that much of the plant damage ascribed to acid rain is actually due to the effects of ozone. Much work needs to be done in the area of sampling. Historical data are lacking and sampling methods are not sufficiently accurate. (JMT)

  10. WASTE ACID DETOXIFICATION AND RECLAMATION

    EPA Science Inventory

    This Environmental Security Technology Certification Program (ESTCP) project demonstrated the Waste Acid Detoxification and Reclamation (WADR) systems ability to recover waste electropolish acid solutions generated during the manufacturing of gun-tubes, and reuse the clean acid. ...

  11. Disorders of Amino Acid Metabolism

    MedlinePlus

    ... Aspiration Syndrome Additional Content Medical News Disorders of Amino Acid Metabolism By Lee M. Sanders, MD, MPH NOTE: ... Metabolic Disorders Disorders of Carbohydrate Metabolism Disorders of Amino Acid Metabolism Disorders of Lipid Metabolism Amino acids are ...

  12. Acid soldering flux poisoning

    MedlinePlus

    The harmful substances in soldering fluxes are called hydrocarbons. They include: Ammonium chloride Rosin Hydrochloric acid Zinc ... Lee DC. Hydrocarbons. In: Marx JA, Hockberger RS, Walls RM, et ... Rosen's Emergency Medicine: Concepts and Clinical Practice . 8th ...

  13. Aminolevulinic Acid Topical

    MedlinePlus

    ... in combination with photodynamic therapy (PDT; special blue light) to treat actinic keratoses (small crusty or scaly ... photosensitizing agents. When aminolevulinic acid is activated by light, it damages the cells of actinic keratosis lesions.

  14. Difficult Decisions: Acid Rain.

    ERIC Educational Resources Information Center

    Miller, John A.; Slesnick, Irwin L.

    1989-01-01

    Discusses some of the contributing factors and chemical reactions involved in the production of acid rain, its effects, and political issues pertaining to who should pay for the clean up. Supplies questions for consideration and discussion. (RT)

  15. Uric acid - urine

    MedlinePlus

    ... to filter fluids and waste normally (chronic glomerulonephritis ) Lead poisoning Long-term (chronic) alcohol use Risks There are ... Elsevier Saunders; 2011:chap 28. Read More Gout Lead poisoning Liver disease Polycythemia vera Uric acid - blood Update ...

  16. Amoxicillin and Clavulanic Acid

    MedlinePlus

    ... is used to treat certain infections caused by bacteria, including infections of the ears, lungs, sinus, skin, ... antibiotics. It works by stopping the growth of bacteria. Clavulanic acid is in a class of medications ...

  17. Hydrofluoric acid poisoning

    MedlinePlus

    Chemical Emergencies: Case Definition: Hydrofluoric Acid . Centers for Disease Control and Prevention, U.S. Dept of Health and Human Services; 2005. Goldfrank LR, ed. Goldfrank's Toxicologic Emergencies . 8th ed. New ...

  18. Lead/acid batteries

    NASA Astrophysics Data System (ADS)

    Bullock, Kathryn R.

    Lead/acid batteries are produced in sizes from less than 1 to 3000 Ah for a wide variety of portable, industrial and automotive applications. Designs include Planté, Fauré or pasted, and tubular electrodes. In addition to the traditional designs which are flooded with sulfuric acid, newer 'valve-regulated" designs have the acid immolibized in a silica gel or absorbed in a porous glass separator. Development is ongoing worldwide to increase the specific power, energy and deep discharge cycle life of this commercially successful system to meet the needs of new applications such as electric vehicles, load leveling, and solar energy storage. The operating principles, current status, technical challenges and commercial impact of the lead/acid battery are reviewed.

  19. Citric acid urine test

    MedlinePlus

    ... used to diagnose renal tubular acidosis and evaluate kidney stone disease. ... tubular acidosis and a tendency to form calcium kidney stones. The following may decrease urine citric acid levels: ...

  20. Pantothenic acid and biotin

    MedlinePlus

    ... JavaScript. Pantothenic acid and biotin are types of B vitamins. They are water-soluble, which means that the ... found in foods that are good sources of B vitamins, including the following: Animal proteins Avocado Broccoli, kale, ...

  1. (Acid rain workshop)

    SciTech Connect

    Turner, R.S.

    1990-12-05

    The traveler presented a paper entitled Susceptibility of Asian Ecosystems to Soil-Mediated Acid Rain Damage'' at the Second Workshop on Acid Rain in Asia. The workshop was organized by the Asian Institute of Technology (Bangkok, Thailand), Argonne National Laboratory (Argonne, Illinois), and Resource Management Associates (Madison, Wisconsin) and was sponsored by the US Department of Energy, the United Nations Environment Program, the United Nations Economic and Social Commission for Asia and the Pacific, and the World Bank. Papers presented on the first day discussed how the experience gained with acid rain in North America and Europe might be applied to the Asian situation. Papers describing energy use projections, sulfur emissions, and effects of acid rain in several Asian countries were presented on the second day. The remaining time was allotted to discussion, planning, and writing plans for a future research program.

  2. Stomach acid test

    MedlinePlus

    Gastric acid secretion test ... The test is done after you have not eaten for a while so fluid is all that remains in ... injected into your body. This is done to test the ability of the cells in the stomach ...

  3. Deoxycholic Acid Injection

    MedlinePlus

    ... severe submental fat ('double chin'; fatty tissue located under the chin). Deoxycholic acid injection is in a ... as a liquid to be injected subcutaneously (just under the skin) by a doctor. Your doctor will ...

  4. Amino Acids and Chirality

    NASA Technical Reports Server (NTRS)

    Cook, Jamie E.

    2012-01-01

    Amino acids are among the most heavily studied organic compound class in carbonaceous chondrites. The abundance, distributions, enantiomeric compositions, and stable isotopic ratios of amino acids have been determined in carbonaceous chondrites fi'om a range of classes and petrographic types, with interesting correlations observed between these properties and the class and typc of the chondritcs. In particular, isomeric distributions appear to correlate with parent bodies (chondrite class). In addition, certain chiral amino acids are found in enantiomeric excess in some chondrites. The delivery of these enantiomeric excesses to the early Earth may have contributed to the origin of the homochirality that is central to life on Earth today. This talk will explore the amino acids in carbonaceous chondritcs and their relevance to the origin of life.

  5. Valproic Acid and Pregnancy

    MedlinePlus

    ... in the treatment of epilepsy, and to treat bipolar disorder and migraines. I have been taking valproic acid ... that women with seizure disorders and women with bipolar disorder might have menstrual problems and difficulty getting pregnant. ...

  6. Amino Acid Metabolism Disorders

    MedlinePlus

    Metabolism is the process your body uses to make energy from the food you eat. Food is ... One group of these disorders is amino acid metabolism disorders. They include phenylketonuria (PKU) and maple syrup ...

  7. Uric acid - blood

    MedlinePlus

    ... High levels of uric acid can sometimes cause gout or kidney disease. You may have this test ... Alcoholism Chemotherapy-related side effects Diabetes Excessive exercise Gout Hypoparathyroidism Lead poisoning Leukemia Medullary cystic kidney disease ...

  8. Citric acid urine test

    MedlinePlus

    ... The test is used to diagnose renal tubular acidosis and evaluate kidney stone disease. Normal Results The ... level of citric acid may mean renal tubular acidosis and a tendency to form calcium kidney stones. ...

  9. Folic Acid Quiz

    MedlinePlus

    ... more easily than natural food folate. Close × Answer: D CORRECT: Folic acid reduces the risk for spina ... g., orange juice and green vegetables). Close × Answer: D CORRECT: Spina bifida and anencephaly are neural tube ...

  10. Folic acid in diet

    MedlinePlus

    ... green leafy vegetables Dried beans and peas (legumes) Citrus fruits and juices Fortified means that vitamins have ... A.D.A.M. Editorial team. Related MedlinePlus Health Topics Folic Acid Browse the Encyclopedia A.D. ...

  11. Amoxicillin and Clavulanic Acid

    MedlinePlus

    ... Amoxicillin is in a class of medications called penicillin-like antibiotics. It works by stopping the growth ... allergic to amoxicillin (Amoxil, Trimox, Wymox), clavulanic acid, penicillin, cephalosporins, or any other medications.tell your doctor ...

  12. Boric Acid Poisoning

    PubMed Central

    Wong, L. C.; Heimbach, M. D.; Truscott, D. R.; Duncan, B. D.

    1964-01-01

    Boric acid poisoning in 11 infants, occurring in the newborn nursery as a result of the accidental and inadvertent use of 2.5% boric acid in the preparation of the formulae, is reported. Five of the infants died. All except two exhibited the classical symptomatology of acute boric acid poisoning, namely, diarrhea, vomiting, erythema, exfoliation, desquamation of the skin, and marked central nervous system irritation. Early manifestations of poisoning were nonspecific, and one patient died before skin manifestations were noted. Peritoneal dialysis, instituted in nine cases, was found to be the most effective method of treatment. It is recommended that boric acid, which is of doubtful therapeutic value, should be completely removed from hospitals, dispensaries and pharmacopoeias. ImagesFig. 1Fig. 2 PMID:14166459

  13. Polymers for acid thickening

    SciTech Connect

    Dixon, K.W.

    1980-09-30

    Acids, thickened with branched emulsion or suspension polymers of diallyldimethylammonium chloride are useful as oil well drilling and fracturing fluids for stimulating well production and in other applications, such as thickeners for cosmetics, paints, adhesives, textiles and printing inks.

  14. Acid-base chemistry

    SciTech Connect

    Hand, C.W.; Blewit, H.L.

    1985-01-01

    The book is not a research compendium and there are no references to the literature. It is a teaching text covering the entire range of undergraduate subject matter dealing with acid-base chemistry (some of it remotely) as taught in inorganic, analytical, and organic chemistry courses. The excellent chapters VII through IX deal in detail with the quantitative aspects of aqueous acid-base equilibria (salt hydrolysis and buffer, titrations, polyprotic and amphoteric substances).

  15. Utilization of acid tars

    SciTech Connect

    Frolov, A.F.; Denisova, T.L.; Aminov, A.N.

    1987-01-01

    Freshly produced acid tar (FPAT), obtained as refinery waste in treating petroleum oils with sulfuric acid and oleum, contains 80% or more sulfuric acid. Of such tars, pond acid tars, which contain up to 80% neutral petroleum products and sulfonated resins, are more stable, and have found applications in the production of binders for paving materials. In this article the authors are presenting results obtained in a study of the composition and reactivity of FPAT and its stability in storage in blends with asphalts obtained in deasphalting operations, and the possibility of using the FPAT in road construction has been examined. In this work, wastes were used which were obtained in treating the oils T-750, KhF-12, I-8A, and MS-14. Data on the change in group chemical composition of FPAT are shown, and the acidity, viscosity, needle penetration, and softening point of acid tars obtained from different grades of oils are plotted as functions of the storage time. It is also shown that the fresh and hardened FPATs differ in their solubilities in various solvents.

  16. Mammalian Fatty Acid Elongases

    PubMed Central

    Jump, Donald B.

    2009-01-01

    Summary Very long chain fatty acids confer functional diversity on cells by variations in their chain length and degree of unsaturation. Microsomal fatty acid elongation represents the major pathway for determining the chain length of saturated, monounsaturated, and polyunsaturated fatty acids in cellular lipids. The overall reaction for fatty acid elongation involves four enzymes and utilizes malonyl CoA, NADPH, and fatty acyl CoA as substrates. While the fundamental pathway and its requirements have been known for many years, recent advances have revealed a family of enzymes involved in the first step of the reaction, i.e., the condensation reaction. Seven fatty acid elongase subtypes (Elovl #1–7) have been identified in the mouse, rat, and human genomes. These enzymes determine the rate of overall fatty acid elongation. Moreover, these enzymes also display differential substrate specificity, tissue distribution, and regulation, making them important regulators of cellular lipid composition as well as specific cellular functions. Herein, methods are described to measure elongase activity, analyze elongation products, and alter cellular elongase expression. PMID:19763486

  17. Neutron Nucleic Acid Crystallography.

    PubMed

    Chatake, Toshiyuki

    2016-01-01

    The hydration shells surrounding nucleic acids and hydrogen-bonding networks involving water molecules and nucleic acids are essential interactions for the structural stability and function of nucleic acids. Water molecules in the hydration shells influence various conformations of DNA and RNA by specific hydrogen-bonding networks, which often contribute to the chemical reactivity and molecular recognition of nucleic acids. However, X-ray crystallography could not provide a complete description of structural information with respect to hydrogen bonds. Indeed, X-ray crystallography is a powerful tool for determining the locations of water molecules, i.e., the location of the oxygen atom of H2O; however, it is very difficult to determine the orientation of the water molecules, i.e., the orientation of the two hydrogen atoms of H2O, because X-ray scattering from the hydrogen atom is very small.Neutron crystallography is a specialized tool for determining the positions of hydrogen atoms. Neutrons are not diffracted by electrons, but are diffracted by atomic nuclei; accordingly, neutron scattering lengths of hydrogen and its isotopes are comparable to those of non-hydrogen atoms. Therefore, neutron crystallography can determine both of the locations and orientations of water molecules. This chapter describes the current status of neutron nucleic acid crystallographic research as well as the basic principles of neutron diffraction experiments performed on nucleic acid crystals: materials, crystallization, diffraction experiments, and structure determination. PMID:26227050

  18. Autophagy on acid.

    PubMed

    Wojtkowiak, Jonathan W; Gillies, Robert J

    2012-11-01

    The microenvironment of solid tumors tends to be more acidic (6.5-7.0) than surrounding normal (7.2-7.4) tissue. Chaotic vasculature, oxygen limitation and major metabolic changes all contribute to the acidic microenvironment. We have previously proposed that low extracellular pH (pHe) plays a critical role in the development and progression of solid tumors. While extracellular acidosis is toxic to most normal cells, cancer cells can adapt and survive under this harsh condition. In this study, we focused on identifying survival strategies employed by cancer cells when challenged with an acidic pHe (6.6-6.7) either acutely or for many generations. While acutely acidic cells did not grow, those acclimated over many generations grew at the same rate as control cells. We observed that these cells induce autophagy in response to acidosis both acutely and chronically, and that this adaptation appears to be necessary for survival. Inhibition of autophagy in low pH cultured cells results in cell death. Histological analysis of tumor xenografts reveals a strong correlation of LC3 protein expression in regions projected to be acidic. Furthermore, in vivo buffering experiments using sodium bicarbonate, previously shown to raise extracellular tumor pH, decreases LC3 protein expression in tumor xenografts. These data imply that autophagy can be induced by extracellular acidosis and appears to be chronically employed as a survival adaptation to acidic microenvironments. PMID:22874557

  19. Method for isolating nucleic acids

    DOEpatents

    Hurt, Jr., Richard Ashley; Elias, Dwayne A.

    2015-09-29

    The current disclosure provides methods and kits for isolating nucleic acid from an environmental sample. The current methods and compositions further provide methods for isolating nucleic acids by reducing adsorption of nucleic acids by charged ions and particles within an environmental sample. The methods of the current disclosure provide methods for isolating nucleic acids by releasing adsorbed nucleic acids from charged particles during the nucleic acid isolation process. The current disclosure facilitates the isolation of nucleic acids of sufficient quality and quantity to enable one of ordinary skill in the art to utilize or analyze the isolated nucleic acids for a wide variety of applications including, sequencing or species population analysis.

  20. Acidification and Acid Rain

    NASA Astrophysics Data System (ADS)

    Norton, S. A.; Veselã½, J.

    2003-12-01

    Air pollution by acids has been known as a problem for centuries (Ducros, 1845; Smith, 1872; Camuffo, 1992; Brimblecombe, 1992). Only in the mid-1900s did it become clear that it was a problem for more than just industrially developed areas, and that precipitation quality can affect aquatic resources ( Gorham, 1955). The last three decades of the twentieth century saw tremendous progress in the documentation of the chemistry of the atmosphere, precipitation, and the systems impacted by acid atmospheric deposition. Chronic acidification of ecosystems results in chemical changes to soil and to surface waters and groundwater as a result of reduction of base cation supply or an increase in acid (H+) supply, or both. The most fundamental changes during chronic acidification are an increase in exchangeable H+ or Al3+ (aluminum) in soils, an increase in H+ activity (˜concentration) in water in contact with soil, and a decrease in alkalinity in waters draining watersheds. Water draining from the soil is acidified and has a lower pH (=-log [H+]). As systems acidify, their biotic community changes.Acidic surface waters occur in many parts of the world as a consequence of natural processes and also due to atmospheric deposition of strong acid (e.g., Canada, Jeffries et al. (1986); the United Kingdom, Evans and Monteith (2001); Sweden, Swedish Environmental Protection Board (1986); Finland, Forsius et al. (1990); Norway, Henriksen et al. (1988a); and the United States (USA), Brakke et al. (1988)). Concern over acidification in the temperate regions of the northern hemisphere has been driven by the potential for accelerating natural acidification by pollution of the atmosphere with acidic or acidifying compounds. Atmospheric pollution ( Figure 1) has resulted in an increased flux of acid to and through ecosystems. Depending on the ability of an ecosystem to neutralize the increased flux of acidity, acidification may increase only imperceptibly or be accelerated at a rate that