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  1. THI1, a Thiamine Thiazole Synthase, Interacts with Ca2+-Dependent Protein Kinase CPK33 and Modulates the S-Type Anion Channels and Stomatal Closure in Arabidopsis.

    PubMed

    Li, Chun-Long; Wang, Mei; Wu, Xiao-Meng; Chen, Dong-Hua; Lv, Hong-Jun; Shen, Jian-Lin; Qiao, Zhu; Zhang, Wei

    2016-02-01

    Thiamine is required for both plant growth and development. Here, the involvement of a thiamine thiazole synthase, THI1, has been demonstrated in both guard cell abscisic acid (ABA) signaling and the drought response in Arabidopsis (Arabidopsis thaliana). THI1 overexpressors proved to be more sensitive to ABA than the wild type with respect to both the activation of guard cell slow type anion channels and stomatal closure; this effectively reduced the rate of water loss from the plant and thereby enhanced its level of drought tolerance. A yeast two-hybrid strategy was used to screen a cDNA library from epidermal strips of leaves for THI1 regulatory factors, and identified CPK33, a Ca(2+)-dependent protein kinase, as interactor with THI1 in a plasma membrane-delimited manner. Loss-of-function cpk33 mutants were hypersensitive to ABA activation of slow type anion channels and ABA-induced stomatal closure, while the CPK33 overexpression lines showed opposite phenotypes. CPK33 kinase activity was essential for ABA-induced stomatal closure. Consistent with their contrasting regulatory role over stomatal closure, THI1 suppressed CPK33 kinase activity in vitro. Together, our data reveal a novel regulatory role of thiamine thiazole synthase to kinase activity in guard cell signaling. PMID:26662273

  2. THI1, a Thiamine Thiazole Synthase, Interacts with Ca2+-Dependent Protein Kinase CPK33 and Modulates the S-Type Anion Channels and Stomatal Closure in Arabidopsis1[OPEN

    PubMed Central

    Li, Chun-Long; Wang, Mei; Wu, Xiao-Meng; Chen, Dong-Hua; Lv, Hong-Jun; Shen, Jian-Lin; Qiao, Zhu; Zhang, Wei

    2016-01-01

    Thiamine is required for both plant growth and development. Here, the involvement of a thiamine thiazole synthase, THI1, has been demonstrated in both guard cell abscisic acid (ABA) signaling and the drought response in Arabidopsis (Arabidopsis thaliana). THI1 overexpressors proved to be more sensitive to ABA than the wild type with respect to both the activation of guard cell slow type anion channels and stomatal closure; this effectively reduced the rate of water loss from the plant and thereby enhanced its level of drought tolerance. A yeast two-hybrid strategy was used to screen a cDNA library from epidermal strips of leaves for THI1 regulatory factors, and identified CPK33, a Ca2+-dependent protein kinase, as interactor with THI1 in a plasma membrane-delimited manner. Loss-of-function cpk33 mutants were hypersensitive to ABA activation of slow type anion channels and ABA-induced stomatal closure, while the CPK33 overexpression lines showed opposite phenotypes. CPK33 kinase activity was essential for ABA-induced stomatal closure. Consistent with their contrasting regulatory role over stomatal closure, THI1 suppressed CPK33 kinase activity in vitro. Together, our data reveal a novel regulatory role of thiamine thiazole synthase to kinase activity in guard cell signaling. PMID:26662273

  3. HMP Binding Protein ThiY and HMP-P Synthase THI5 Are Structural Homologues

    SciTech Connect

    Bale, Shridhar; Rajashankar, Kanagalaghatta R.; Perry, Kay; Begley, Tadhg P.; Ealick, Steven E.

    2010-10-14

    The ATP-binding cassette transporter system ThiXYZ transports N-formyl-4-amino-5-(aminomethyl)-2-methylpyrimidine (FAMP), a thiamin salvage pathway intermediate, into cells. FAMP is then converted to 4-amino-5-(hydroxymethyl)-2-methylpyrimidine (HMP) and recycled into the thiamin biosynthetic pathway. ThiY is the periplasmic substrate binding protein of the ThiXYZ system and delivers the substrate FAMP to the transmembrane domain. We report the crystal structure of Bacillus halodurans ThiY with FAMP bound at 2.4 {angstrom} resolution determined by single-wavelength anomalous diffraction phasing. The crystal structure reveals that ThiY belongs to the group II periplasmic binding protein family. The closest structural homologues of ThiY are periplasmic binding proteins involved in alkanesulfonate/nitrate and bicarbonate transport. ThiY is also structurally homologous to thiamin binding protein (TbpA) and to thiaminase-I. THI5 is responsible for the synthesis of 4-amino-5-(hydroxymethyl)-2-methylpyrimidine phosphate in the thiamin biosynthetic pathway of eukaryotes and is approximately 25% identical in sequence with ThiY. A homology model of Saccharomyces cerevisiae THI5 was generated on the basis of the structure of ThiY. Many features of the thiamin pyrimidine binding site are shared between ThiY and THI5, suggesting a common ancestor.

  4. Structure of trifunctional THI20 from yeast

    SciTech Connect

    French, Jarrod B.; Begley, Tadhg P.; Ealick, Steven E.

    2011-08-24

    In a recently characterized thiamin-salvage pathway, thiamin-degradation products are hydrolyzed by thiaminase II, yielding 4-amino-5-hydroxymethyl-2-methylpyrimidine (HMP). This compound is an intermediate in thiamin biosynthesis that, once phosphorylated by an HMP kinase, can be used to synthesize thiamin monophosphate. Here, the crystal structure of Saccharomyces cerevisiae THI20, a trifunctional enzyme containing an N-terminal HMP kinase/HMP-P kinase (ThiD-like) domain and a C-terminal thiaminase II (TenA-like) domain, is presented. Comparison to structures of the monofunctional enzymes reveals that while the ThiD-like dimer observed in THI20 resembles other ThiD structures, the TenA-like domain, which is tetrameric in all previously reported structures, forms a dimer. Similarly, the active site of the ThiD-like domain of THI20 is highly similar to other known ThiD enzymes, while the TenA-like active site shows unique features compared with previously structurally characterized TenAs. In addition, a survey of known TenA structures revealed two structural classes, both of which have distinct conserved features. The TenA domain of THI20 possesses some features of both classes, consistent with its ability to hydrolyze both thiamin and the thiamin-degradation product 2-methyl-4-amino-5-aminomethylpyrimidine.

  5. Structure of trifunctional THI20 from yeast

    PubMed Central

    French, Jarrod B.; Begley, Tadhg P.; Ealick, Steven E.

    2011-01-01

    In a recently characterized thiamin-salvage pathway, thiamin-degradation products are hydrolyzed by thiaminase II, yielding 4-amino-5-hydroxymethyl-2-methylpyrimidine (HMP). This compound is an intermediate in thiamin biosynthesis that, once phosphorylated by an HMP kinase, can be used to synthesize thiamin monophosphate. Here, the crystal structure of Saccharomyces cerevisiae THI20, a trifunctional enzyme containing an N-terminal HMP kinase/HMP-P kinase (ThiD-like) domain and a C-terminal thia­min­ase II (TenA-like) domain, is presented. Comparison to structures of the monofunctional enzymes reveals that while the ThiD-like dimer observed in THI20 resembles other ThiD structures, the TenA-like domain, which is tetrameric in all previously reported structures, forms a dimer. Similarly, the active site of the ThiD-like domain of THI20 is highly similar to other known ThiD enzymes, while the TenA-like active site shows unique features compared with previously structurally characterized TenAs. In addition, a survey of known TenA structures revealed two structural classes, both of which have distinct conserved features. The TenA domain of THI20 possesses some features of both classes, consistent with its ability to hydrolyze both thiamin and the thiamin-degradation product 2-methyl-4-amino-5-aminomethylpyrimidine. PMID:21904031

  6. Bleomycin-Treated Chimeric Thy1-Deficient Mice with Thy1-Deficient Myofibroblasts and Thy-Positive Lymphocytes Resolve Inflammation without Affecting the Fibrotic Response

    PubMed Central

    Cohen, Pazit Y.; Breuer, Raphael; Zisman, Philip; Wallach-Dayan, Shulamit B.

    2015-01-01

    Lung fibrosis is characterized by abnormal accumulation of fibroblasts in the interstitium of the alveolar space. Two populations of myofibroblasts, distinguished by Thy1 expression, are detected in human and murine lungs. Accumulation of Thy1-negative (Thy1−) myofibroblasts was shown in the lungs of humans with idiopathic pulmonary fibrosis (IPF) and of bleomycin-treated mice. We aimed to identify genetic changes in lung myofibroblasts following Thy1 crosslinking and assess the impact of specific lung myofibroblast Thy1-deficiency, in vivo, in bleomycin-injured mouse lungs. Thy1 increased in mouse lung lymphocytes following bleomycin injury but decreased in myofibroblasts when fibrosis was at the highest point (14 days), as assessed by immunohistochemistry. Using gene chip analysis, we detected that myofibroblast Thy1 crosslinking mediates downregulation of genes promoting cell proliferation, survival, and differentiation, and reduces production of extracellular matrix (ECM) components, while concurrently mediating the upregulation of genes known to foster inflammation and immunological functions. Chimeric Thy1-deficient mice with Thy1+ lymphocytes and Thy1− myofibroblasts showed fibrosis similar to wild-type mice and an increased number of CD4/CD25 regulatory T cells, with a concomitant decrease in inflammation. Lung myofibroblasts downregulate Thy1 expression to increase their proliferation but to diminish the in vivo inflammatory milieu. Inflammation is not essential for evolution of fibrosis as was previously stated. PMID:26300593

  7. Imaging Acute Neuromuscular Explants from Thy1 Mouse Lines.

    PubMed

    Marinković, Petar; Godinho, Leanne; Misgeld, Thomas

    2015-09-01

    Because core facilities that generate transgenic founder mice for a reasonable fee are now available at most major research institutions, generating new Thy1-XFP transgenic animals (in which XFP stands for any fluorescent protein) is an option even for relatively small laboratories. Here, we provide a protocol for screening offspring of Thy1 transgenic founders. Acute neuromuscular explants are obtained from 3-wk-old F1 mice that have been produced by crossing Thy1 transgenic founders and commercially obtained inbred mice. Thy1-driven expression is detected by fluorescence microscopy. PMID:26330628

  8. Generation of Tissue Sections for Screening Thy1 Mouse Lines.

    PubMed

    Marinković, Petar; Godinho, Leanne; Misgeld, Thomas

    2015-09-01

    New generations of Thy1-XFP transgenic mice (where XFP stands for any fluorescent protein) can now be readily generated, given the availability of core facilities or commercial providers of Thy1 pronuclear injections. Here, we provide a protocol for screening founder progeny. Transcardial perfusion is performed on 3-wk-old F1 mice that have been produced by crossing Thy1 transgenic founders and commercially obtained inbred mice. Cryosections are generated, and Thy1-driven expression is detected by histological characterization. PMID:26330627

  9. Tu-144LL Factory scenes

    NASA Technical Reports Server (NTRS)

    1995-01-01

    A jointly funded activity by the NASA High Speed Research (HSR) program and the Boeing Commercial Airplane Group took place to obtain experimental flight data on the Tu-144 supersonic transport built by Russia. The Tu-144 was modified by the Tupolev Aircraft Design Bureau, Moscow, Russia, in 1995-1996 into the Tu-144LL Flying Laboratory to perform flight experiments as part of the NASA HSR Program. Knowledge gained from the flights will benefit the NASA efforts to develop the technology that may enable design of an efficient, environmentally friendly second-generation supersonic transport in this country. This program involved eight experiments -- six aboard the aircraft and two ground test engine experiments. Between November 1996 and February 1998 the Tu-144LL flew 19 research flights. The follow-on Tu-144LL program encompassed about eight flights, focusing on extensions of five experiments from the first project and two new experiments to measure fuel system temperatures and to define in-flight wing deflections. This movie clip runs 1:17 minutes and shows several different views of the Russian Tu-144 LL supersonic transport, on the ramp, in its hangar, a rear view of the engine exhaust nozzles, the avionics bay, cockpit and finally a shot looking forward along the fuselage from the tail.

  10. Generation of Thy1 Constructs for Pronuclear Injection.

    PubMed

    Marinković, Petar; Godinho, Leanne; Misgeld, Thomas

    2015-10-01

    With easy access to core facilities or commercial providers of pronuclear injections, generating simple Thy1-XFP transgenic mice (where XFP stands for any fluorescent protein) is now a possibility even for small laboratories. The generation of new Thy1 transgenic lines generally consists of five steps: (1) engineering and characterization of the desired fluorescent reporter protein, (2) cloning of the reporter protein into the Thy1 vector, (3) linearization and purification of the new Thy1 construct, (4) pronuclear injection to generate founders, and (5) screening of founder progeny to establish transgenic lines. Here, we provide a protocol for Steps 2 and 3. The sequence for a desired fluorescent reporter protein is cloned into the XhoI restriction site of the Thy1 vector. This usually involves blunt-end cloning because the traditional Thy1 vector does not carry an intact multiple cloning site. Following successful cloning, the DNA is prepared for pronuclear injection by linearizing it using EcoRI and PvuI restriction enzymes. The purified linearized DNA must then be sent to a facility specializing in pronuclear injection to generate transgenic founder mice. PMID:26430258

  11. Conditional Antifolate Resistance in Bacillus subtilis thyA

    PubMed Central

    Farmer, James L.

    1979-01-01

    Resistance to antifolates in Bacillus subtilis strains results from the presence of an antifolate resistance mutation (afo). Strains which are thyA+afo are unconditionally resistant to antifolates. The conditional resistance of thyA afo strains is hypothesized to be due to the thyB+ gene product (thymidylate synthetase B) having a high Km for the folate substrate, thus leading to thymineless death in the presence of antifolates. An alternative model for conditional antifolate resistance was shown to be incorrect by analysis of folate metabolism in methotrexate-treated cells. Genetic analysis and studies of the response of afo+ cells to methotrexate suggested that most, if not all, B. subtilis thymine-requiring mutants are afo. Analysis of dihydrofolate reductase from afo cells did not reveal an obvious mechanism for antifolate resistance in those cells. PMID:111614

  12. Isolation and characterization of Thy 1 homologue from human thymus.

    PubMed

    Bonewald, L F; Goust, J M; Sade, R M; Wang, A C

    1985-01-01

    A 40 000 M.W. glycoprotein was isolated from human thymus. This molecule binds lentil lectin, reacts with an antiserum made against the p25 antigen (the human Thy 1 homologue) and possesses almost identical amino acid composition as the p25 antigen and its 40 000 M.W. dimer. PMID:2864756

  13. Thy-1, the enigmatic extrovert on the neuronal surface.

    PubMed

    Morris, R

    1992-10-01

    Thy-1 is a small glycoprotein of 110 amino acids which, folded in the characteristic structure of an immunoglobulin variable domain, are enchored to the plasma membrane via a glycophosphatidylinositol (GPI) tail (Fig. 1). It is a major component of the surface of various cell types, including neurons, at certain stages of their development. These qualities doubtlessly appeal to certain cognoscenti, but it is not clear why they would raise Thy-1 to the status of a favourite molecule. Indeed, few scientists readily admit to having a favourite. We study individual molecules because science is rooted in specific observations; but we do so in order to discover mechanisms of general importance. A molecule's appeal is dependent on its ability to reveal novel aspects of how nature works. Thy-1 has been unusual in this respect. It was the first lymphocyte surface antigen shown to be restricted to a functional subset of lymphocytes (T cells in the mouse), a finding crucial to the development of cellular immunology; it was one of the first cell surface molecules to be sequenced and indicated the importance of immunoglobulin domains and GPI anchors as structural motifs; it has been pivotal in studies demonstrating that GPI-anchored molecules are able to signal across the membrane they do not span. Thy-1 has revealed this much, however, with the charm of an adroit stripper: it has always promised glimpses of things more exciting than that displayed. In particular, the function of this molecule has never emerged.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:1285421

  14. Thermal stability and binding energetics of thymidylate synthase ThyX.

    PubMed

    Krumova, Sashka; Todinova, Svetla; Tileva, Milena; Bouzhir-Sima, Latifa; Vos, Marten H; Liebl, Ursula; Taneva, Stefka G

    2016-10-01

    The bacterial thymidylate synthase ThyX is a multisubstrate flavoenzyme that takes part in the de novo synthesis of thymidylate in a variety of microorganisms. Herein we study the effect of FAD and dUMP binding on the thermal stability of wild type (WT) ThyX from the mesophilic Paramecium bursaria chlorella virus-1 (PBCV-1) and from the thermophilic bacterium Thermotoga maritima (TmThyX), and from two variants of TmThyX, Y91F and S88W, using differential scanning calorimetry. The energetics underlying these processes was characterized by isothermal titration calorimetry. The PBCV-1 protein is significantly less stable against the thermal challenge than the TmThyX WT. FAD exerted stabilizing effect greater for PBCV-1 than for TmThyX and for both mutants, whereas binding of dUMP to FAD-loaded proteins stabilized further only TmThyX. Different thermodynamic signatures describe the FAD binding to the WT ThyX proteins. While TmThyX binds FAD with a low μM binding affinity in a process characterized by a favorable entropy change, the assembly of PBCV-1 with FAD is governed by a large enthalpy change opposed by an unfavorable entropy change resulting in a relatively strong nM binding. An enthalpy-driven formation of a high affinity ternary ThyX/FAD/dUMP complex was observed only for TmThyX. PMID:27268384

  15. Tu-144LL ramp taxi and takeoff

    NASA Technical Reports Server (NTRS)

    1998-01-01

    A jointly funded activity by the NASA High Speed Research (HSR) program and the Boeing Commercial Airplane Group took place to obtain experimental flight data on the Tu-144 supersonic transport built by Russia. The Tu-144 was modified by the Tupolev Aircraft Design Bureau, Moscow, Russia, in 1995-1996 into the Tu-144LL Flying Laboratory to perform flight experiments as part of the NASA HSR Program. Knowledge gained from the flights will benefit the NASA efforts to develop technology that may enable design of an efficient, environmentally friendly second-generation supersonic transport in this country. This program involved eight experiments -- six aboard the aircraft and two ground test engine experiments. Between November 1996 and February 1998 the Tu-144LL flew 19 research flights. The follow-on Tu-144LL program encompassed about eight flights, focusing on extensions of five experiments from the first project and two new experiments to measure fuel system temperatures and to define in-flight wing deflections. This 31-second clip shows the Russian Tu-144 LL supersonic transport on the ramp in Moscow, then taxiing into position and making its takeoff run, rotating from the runway and climbing away.

  16. Anti-Proliferative Actions of T-Type Calcium Channel Inhibition in Thy1 Nephritis

    PubMed Central

    Cove-Smith, Andrea; Mulgrew, Christopher J.; Rudyk, Olena; Dutt, Neelanjana; McLatchie, Linda M.; Shattock, Michael J.; Hendry, Bruce M.

    2014-01-01

    Aberrant proliferation of mesangial cells (MCs) is a key finding in progressive glomerular disease. TH1177 is a small molecule that has been shown to inhibit low-voltage activated T-type Ca2+ channels (TCCs). The current study investigates the effect of TH1177 on MC proliferation in vitro and in vivo. The effect of Ca2+ channel inhibition on primary rat MC proliferation in vitro was studied using the microculture tetrazolium assay and by measuring bromodeoxyuridine incorporation. In vivo, rats with Thy1 nephritis were treated with TH1177 or vehicle. Glomerular injury and average glomerular cell number were determined in a blinded fashion. Immunostaining for Ki-67 and phosphorylated ERK were also performed. The expression of TCC isoforms in healthy and diseased tissue was investigated using quantitative real-time PCR. TCC blockade caused a significant reduction in rat MC proliferation in vitro, whereas L-type inhibition had no effect. Treatment of Thy1 nephritis with TH1177 significantly reduced glomerular injury (P < 0.005) and caused a 49% reduction in glomerular cell number (P < 0.005) compared to the placebo. TH1177 also reduced Ki-67-positive and pERK-positive cells per glomerulus by 52% (P < 0.01 and P < 0.005, respectively). These results demonstrate that TH1177 inhibits MC proliferation in vitro and in vivo, supporting the hypothesis that TCC inhibition may be a useful strategy for studying and modifying MC proliferative responses to injury. PMID:23746655

  17. Tissue-specific control elements of the Thy-1 gene.

    PubMed Central

    Vidal, M; Morris, R; Grosveld, F; Spanopoulou, E

    1990-01-01

    We have exploited the structural homology, but different patterns of expression of the murine and human Thy-1 genes to map a number of tissue-specific enhancer elements in the genes. All of these are located downstream from the site of transcriptional initiation. The human gene contains separate elements which direct expression to the kidney or spleen epithelium. The murine gene lacks these elements but instead contains a thymocyte specific enhancer in the third intron. Developmentally-regulated expression in nerve cells is directed (at least in part) by an atypical element in the first intron. The latter is active on heterologous promoters, but is position and distance dependent. Images Fig. 2. Fig. 4. Fig. 5. Fig. 6. PMID:1968831

  18. Thy-1+ dendritic cells in murine epidermis are bone marrow-derived

    SciTech Connect

    Breathnach, S.M.; Katz, S.I.

    1984-07-01

    Thy-1+, Ly-5+ dendritic cells have recently been described as a resident cell population in murine epidermis, but their ontogeny and function are unknown. The origin and turnover of epidermal Thy-1+ cells utilizing chimeric mice were investigated. Lethally x-irradiated AKR/J (Thy-1.1+) and AKR/Cum (Thy-1.2+) mice were reconstituted with allogeneic bone marrow cells with or without thymocytes from congenic AKR/Cum or AKR/J mice, respectively. The density of residual indigenous Thy-1.1+ cells in AKR/J chimeras and Thy-1.2+ cells in AKR/Cum chimeras was substantially reduced following x-irradiation, as determined by immunofluorescence staining of epidermal sheets. Epidermal repopulation by allogeneic Thy-1+ dendritic epidermal cells was first observed at 5 weeks in AKR/J chimeras and at 7 weeks in AKR/Cum chimeras and progressed slowly. Repopulation was not enhanced by increasing the number of allogeneic bone marrow cells injected from 2 X 10(7) to 10(8) cells or by the addition of 8 X 10(7) allogeneic thymocytes to the donor inoculate. Epidermal repopulation by allogeneic Thy-1.2+ cells was not seen in AKR/J mice reconstituted with syngeneic bone marrow cells and allogeneic Thy-1.2+ AKR/Cum thymocytes. Taken together, these results indicate that Thy-1+ dendritic epidermal cells are derived from the bone marrow and suggest that they are not related to conventional peripheral T-lymphocytes.

  19. Component of Caramel Food Coloring, THI, Causes Lymphopenia Indirectly via a Key Metabolic Intermediate.

    PubMed

    Ohtoyo, Mamoru; Machinaga, Nobuo; Inoue, Ryotaku; Hagihara, Katsunobu; Yuita, Hiroshi; Tamura, Masakazu; Hashimoto, Ryuji; Chiba, Jun; Muro, Fumihito; Watanabe, Jun; Kobayashi, Yoshimasa; Abe, Koji; Kita, Yasuo; Nagasaki, Miyuki; Shimozato, Takaichi

    2016-05-19

    Caramel color is widely used in the food industry, and its many variations are generally considered to be safe. It has been known for a long time that THI (2-acetyl-4-(tetrahydroxybutyl)imidazole), a component of caramel color III, causes lymphopenia in animals through sphingosine 1-phosphate (S1P) lyase (S1PL) inhibition. However, this mechanism of action has not been fully validated because THI does not inhibit S1PL in vitro. To reconcile this situation, we examined molecular details of THI mechanism of action using "smaller" THI derivatives. We identified a bioactive derivative, A6770, which has the same lymphopenic effect as THI via S1PL inhibition. In the case of A6770 we observe this effect both in vitro and in vivo, and demonstrate that A6770 is phosphorylated and inhibits S1PL in the same way as 4-deoxypyridoxine. In addition, A6770 was detected in rat plasma following oral administration of THI, suggesting that A6770 is a key metabolic intermediate of THI. PMID:27185637

  20. Domain Organization in Candida glabrata THI6, a Bifunctional Enzyme Required for Thiamin Biosynthesis in Eukaryotes

    SciTech Connect

    Paul, Debamita; Chatterjee, Abhishek; Begley, Tadhg P.; Ealick, Steven E.

    2010-11-15

    THI6 is a bifunctional enzyme found in the thiamin biosynthetic pathway in eukaryotes. The N-terminal domain of THI6 catalyzes the ligation of the thiamin thiazole and pyrimidine moieties to form thiamin phosphate, and the C-terminal domain catalyzes the phosphorylation of 4-methyl-5-hydroxyethylthiazole in a salvage pathway. In prokaryotes, thiamin phosphate synthase and 4-methyl-5-hydroxyethylthiazole kinase are separate gene products. Here we report the first crystal structure of a eukaryotic THI6 along with several complexes that characterize the active sites responsible for the two chemical reactions. THI6 from Candida glabrata is a homohexamer in which the six protomers form a cage-like structure. Each protomer is composed of two domains, which are structurally homologous to their monofunctional bacterial counterparts. Two loop regions not found in the bacterial enzymes provide interactions between the two domains. The structures of different protein-ligand complexes define the thiazole and ATP binding sites of the 4-methyl-5-hydroxyethylthiazole kinase domain and the thiazole phosphate and 4-amino-5-hydroxymethyl-2-methylpyrimidine pyrophosphate binding sites of the thiamin phosphate synthase domain. Our structural studies reveal that the active sites of the two domains are 40 {angstrom} apart and are not connected by an obvious channel. Biochemical studies show 4-methyl-5-hydroxyethylthiazole phosphate is a substrate for THI6; however, adenosine diphospho-5{beta}-ethyl-4-methylthiazole-2-carboxylic acid, the product of THI4, is not a substrate for THI6. This suggests that an unidentified enzyme is necessary to produce the substrate for THI6 from the THI4 product.

  1. Immunosuppressive effects of 2-acetyl-4-tetrahydroxybutyl imidazole (THI) in the rat.

    PubMed Central

    Gobin, S J; Phillips, J A

    1991-01-01

    THI is a component of ammonia caramel, a widely used food colouring. The effect of THI on the immune system has been determined in the male F344 rat. THI was given in the drinking water at doses of 1, 10 and 50 mg/l (equivalent to 0.1, 1 and 5 mg/kg per day) to animals on a vitamin B6-deficient diet. After 1 week, the immune competence of the animals was assessed under continued THI treatment. No marked changes in thymus or spleen weight were observed after THI treatment, although there was an increased number of pyknotic cells in the thymic cortex, mainly engulfed by macrophages and there appeared to be a slight thinning of the cortex area. THI produced a significant loss in T and B lymphocytes in peripheral blood but not in the spleen. No change in natural killer (NK) cell activity against YAC-1 target cells was observed in the spleen. The observed increase in NK cell activity in peripheral blood was due to an increase in circulating large granular lymphocytes (LGL). Although the serum antibody titre against keyhole limpet haemocyanin (KLH) was not affected by THI treatment, B cells showed less proliferation when cultured with lipopolysaccharide. T cell function was impeded as measured in mitogen-induced proliferation assay, delayed-type hypersensitivity assay and host versus graft (popliteal lymph node) assay. The results indicate that THI is an immunosuppressor in the rat, in whom it can produce profound lymphopenia and suppression of cell-mediated immunity. Images Fig. 1 PMID:1864015

  2. Targeting of Helicobacter pylori thymidylate synthase ThyX by non-mitotoxic hydroxy-naphthoquinones

    PubMed Central

    Skouloubris, Stéphane; Djaout, Kamel; Lamarre, Isabelle; Lambry, Jean-Christophe; Anger, Karine; Briffotaux, Julien; Liebl, Ursula; de Reuse, Hilde; Myllykallio, Hannu

    2015-01-01

    ThyX is an essential thymidylate synthase that is mechanistically and structurally unrelated to the functionally analogous human enzyme, thus providing means for selective inhibition of bacterial growth. To identify novel compounds with anti-bacterial activity against the human pathogenic bacterium Helicobacter pylori, based on our earlier biochemical and structural analyses, we designed a series of eighteen 2-hydroxy-1,4-naphthoquinones (2-OH-1,4-NQs) that target HpThyX. Our lead-like molecules markedly inhibited the NADPH oxidation and 2′-deoxythymidine-5′-monophosphate-forming activities of HpThyX enzyme in vitro, with inhibitory constants in the low nanomolar range. The identification of non-cytotoxic and non-mitotoxic 2-OH-1,4-NQ inhibitors permitted testing their in vivo efficacy in a mouse model for H. pylori infections. Despite the widely assumed toxicity of naphthoquinones (NQs), we identified tight-binding ThyX inhibitors that were tolerated in mice and can be associated with a modest effect in reducing the number of colonizing bacteria. Our results thus provide proof-of-concept that targeting ThyX enzymes is a highly feasible strategy for the development of therapies against H. pylori and a high number of other ThyX-dependent pathogenic bacteria. We also demonstrate that chemical reactivity of NQs does not prevent their exploitation as anti-microbial compounds, particularly when mitotoxicity screening is used to prioritize these compounds for further experimentation. PMID:26040760

  3. Conformational coupling of integrin and Thy-1 regulates Fyn priming and fibroblast mechanotransduction

    PubMed Central

    Fiore, Vincent F.; Strane, Patrick W.; Bryksin, Anton V.; White, Eric S.; Hagood, James S.

    2015-01-01

    Progressive fibrosis is characterized by excessive deposition of extracellular matrix (ECM), resulting in gross alterations in tissue mechanics. Changes in tissue mechanics can further augment scar deposition through fibroblast mechanotransduction. In idiopathic pulmonary fibrosis, a fatal form of progressive lung fibrosis, previous work has shown that loss of Thy-1 (CD90) expression in fibroblasts correlates with regions of active fibrogenesis, thus representing a pathologically relevant fibroblast subpopulation. We now show that Thy-1 is a regulator of fibroblast rigidity sensing. Thy-1 physically couples to inactive αvβ3 integrins via its RGD-like motif, altering baseline integrin avidity to ECM ligands and also facilitating preadhesion clustering of integrin and membrane rafts via Thy-1’s glycophosphatidylinositol tether. Disruption of Thy-1–αvβ3 coupling altered recruitment of Src family kinases to adhesion complexes and impaired mechanosensitive, force-induced Rho signaling, and rigidity sensing. Loss of Thy-1 was sufficient to induce myofibroblast differentiation in soft ECMs and may represent a physiological mechanism important in wound healing and fibrosis. PMID:26459603

  4. Targeting of Helicobacter pylori thymidylate synthase ThyX by non-mitotoxic hydroxy-naphthoquinones.

    PubMed

    Skouloubris, Stéphane; Djaout, Kamel; Lamarre, Isabelle; Lambry, Jean-Christophe; Anger, Karine; Briffotaux, Julien; Liebl, Ursula; de Reuse, Hilde; Myllykallio, Hannu

    2015-06-01

    ThyX is an essential thymidylate synthase that is mechanistically and structurally unrelated to the functionally analogous human enzyme, thus providing means for selective inhibition of bacterial growth. To identify novel compounds with anti-bacterial activity against the human pathogenic bacterium Helicobacter pylori, based on our earlier biochemical and structural analyses, we designed a series of eighteen 2-hydroxy-1,4-naphthoquinones (2-OH-1,4-NQs) that target HpThyX. Our lead-like molecules markedly inhibited the NADPH oxidation and 2'-deoxythymidine-5'-monophosphate-forming activities of HpThyX enzyme in vitro, with inhibitory constants in the low nanomolar range. The identification of non-cytotoxic and non-mitotoxic 2-OH-1,4-NQ inhibitors permitted testing their in vivo efficacy in a mouse model for H. pylori infections. Despite the widely assumed toxicity of naphthoquinones (NQs), we identified tight-binding ThyX inhibitors that were tolerated in mice and can be associated with a modest effect in reducing the number of colonizing bacteria. Our results thus provide proof-of-concept that targeting ThyX enzymes is a highly feasible strategy for the development of therapies against H. pylori and a high number of other ThyX-dependent pathogenic bacteria. We also demonstrate that chemical reactivity of NQs does not prevent their exploitation as anti-microbial compounds, particularly when mitotoxicity screening is used to prioritize these compounds for further experimentation. PMID:26040760

  5. Chloroplast EF-Tu and thermal aggregation of Rubisco activase

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Chloroplast protein synthesis elongation factor, EF-Tu, has been implicated in heat tolerance in maize. The recombinant precursor of this protein, pre-EF-Tu, has been found to exhibit chaperone activity and protect heat-labile proteins, such as citrate synthase and malate dehydrogenase, from therma...

  6. 40 CFR Appendixes T-U to Part 51 - [Reserved

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 2 2010-07-01 2010-07-01 false T Appendixes T-U to Part 51 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS REQUIREMENTS FOR PREPARATION, ADOPTION, AND SUBMITTAL OF IMPLEMENTATION PLANS Appendixes T-U to Part 51...

  7. Anomalous expression of Thy1 (CD90) in B-cell lymphoma cells and proliferation inhibition by anti-Thy1 antibody treatment

    SciTech Connect

    Ishiura, Yoshihito; Kotani, Norihiro; Yamashita, Ryusuke; Yamamoto, Harumi; Kozutsumi, Yasunori; Honke, Koichi

    2010-05-28

    The anti-CD20 monoclonal antibody (Ab) rituximab is accepted to be an effective therapeutic Ab for malignant B-cell lymphoma; however, discovery of other cell surface antigens is required for the option of antibody medicine. Considering that many tumor-associated antigens are glycans, we have searched glycoconjugates for the candidate antigens that therapeutic Abs target. To this end, we first focused on the difference in the glycogenes expression in terms of Epstein-Barr virus (EBV) infection of a Burkitt's lymphoma cell line, Akata. Using DNA array, flow cytometry and Western blotting, we found that Thy1 was highly expressed in EBV-positive Akata cells. Subsequently, Thy1 was found to be expressed in other B-cell lymphoma cell lines: BJAB, MutuI, and MutuIII, irrespective of EBV infection. Treatment of these cells with an anti-Thy1 monoclonal antibody inhibited proliferation more strongly than the therapeutic Ab rituximab. The B-cell lymphoma cell lines were classified based on the extent of the proliferation inhibition, which was not correlated with the expression level of Thy1. It is suggested that stable residence of receptor tyrosine kinases in lipid rafts sustains cell growth in B-cell lymphoma cells.

  8. DIRECT THY-1/αvβ3 INTEGRIN INTERACTION MEDIATES NEURON TO ASTROCYTE COMMUNICATION

    PubMed Central

    Hermosilla, Tamara; Muñoz, Daniel; Herrera-Molina, Rodrigo; Valdivia, Alejandra; Muñoz, Nicolás; Nham, Sang-Uk; Schneider, Pascal; Burridge, Keith; Quest, Andrew F. G.; Leyton, Lisette

    2008-01-01

    Thy-1 is an abundant neuronal glycoprotein of poorly defined function. We recently provided evidence indicating that Thy-1 clusters a β3-containing integrin in astrocytes to induce tyrosine phosphorylation, RhoA activation and the formation of focal adhesions and stress fibers. To date, the α subunit partner of β3 integrin in DI TNC1 astrocytes is unknown. Similarly, the ability of neuronal, membrane-bound Thy-1 to trigger astrocyte signaling via integrin engagement remains speculation. Here, evidence that αv forms an αvβ3 heterodimer in DI TNC1 astrocytes was obtained. In neuron-astrocyte association assays, the presence of either anti-αv or anti-β3 integrin antibodies reduced cell-cell interaction demonstrating the requirement of both integrin subunits for this association. Moreover, anti-Thy-1 antibodies blocked stimulation of astrocytes by neurons but not the binding of these two cell types. Thus, neuron-astrocyte association involved binding between molecular components in addition to the Thy-1-integrin; however, the signaling events leading to focal adhesion formation in astrocytes depended exclusively on the latter interaction. Additionally, wild-type (RLD) but not mutated (RLE) Thy-1 was shown to directly interact with αvβ3 integrin by Surface Plasmon Resonance analysis. This interaction was promoted by divalent cations and was species-independent. Together, these results demonstrate that the αvβ3 integrin heterodimer interacts directly with Thy-1 present on neuronal cells to stimulate astrocytes. PMID:18346467

  9. The SU Ursae Majoris Star TU Crateris

    NASA Astrophysics Data System (ADS)

    Mennickent, R. E.; Patterson, J.; O'Donoghue, D.; Unda, E.; Harvey, D.; Vanmuster, T.; Bolt, G.

    We have established that the cataclysmic variable TU Crateris is an SU UMa star. Superhumps were observed after 3 days of the 1998 March-April supermaximum, repeating with a mean period of P_s = 0d.08535(5). Timings of superhump maxima revealed a period decrease of P = 7.2 x 10^-5. Based on an empirical relationship, we estimated an orbital period of 0d.0810(25), just in the lower limit of the reported quiescence photometric period. Hα emission lines during quiescence vary with the orbital period with radial velocity half amplitude of 70 +/- 19 km s^-1, evidencing a hotspot located in front of the standard position.

  10. Algebraic decomposition of the TU wave morphology patterns.

    PubMed

    Padrini, R; Butrous, G; Camm, A J; Malik, M

    1995-12-01

    In principle, the T wave results from the differences in durations of action potentials (AP) of different ventricular regions. Based on this concept, a mathematical model has been developed that represents the TU wave morphology as a summation of four AP-like functions: TU = S1 - S2 + L1 - L2. The sigmoidal shape of AP-like curves is produced by Hill's equation V(t) = a . tn/(bn + tn). Each of the decomposition functions is characterized by two parameters: the amplitude at the beginning of QRS (Amax), and the duration at 5% of Amax (D95). The set of four decomposition functions leads to eight parameters that provide detailed characteristics of the TU wave morphology. The model was validated using 170 TU wave complexes recorded digitally in leads V2-V6 from 22 normal subjects and 12 patients with abnormal TU wave morphologies (negative, biphasic, and notched T waves). The electrocardiographic signals were sampled at 100 Hz and a best-fit procedure was used to obtain the decomposition. In all cases the coefficients of correlation between original TU patterns and their mathematical models were > or = 0.99. The mean absolute difference between the observed and modeled values of the TU patterns was similar in cases with normal and abnormal TU wave morphologies (4.65 +/- 0.41 microV vs 5.19 +/- 0.48 microV respectively) demonstrating that the model is capable of describing and categorizing various TU patterns by a set of eight numerical parameters. PMID:8771134

  11. Thy-1 Is Expressed in Hepatic Myofibroblasts and Not Oval Cells in Stem Cell-Mediated Liver Regeneration

    PubMed Central

    Dezső, Katalin; Jelnes, Peter; László, Viktória; Baghy, Kornélia; Bödör, Csaba; Paku, Sándor; Tygstrup, Niels; Bisgaard, Hanne Cathrine; Nagy, Peter

    2007-01-01

    Thy-1, a marker of hematopoietic stem cells, has been reported to be expressed by oval cells proliferating during stem cell-mediated regeneration in rat liver, suggesting a relationship between the two cell populations. Consequently, Thy-1 has become an accepted cell surface marker to sort hepatic oval cells. In the present study we used the well-characterized 2-acetylaminfluorene/partial hepatectomy model to induce transit-amplification of hepatic oval cells in the regenerating liver and characterized Thy-1 expression using Northern hybridization, quantitative reverse transcriptase-polymerase chain reaction analysis, immunofluorescence confocal microscopy, and immunoelectronmicroscopy. We found that Thy-1 expression was induced during transit-amplification of the oval cell population, but Thy-1 mRNA was not present in the α-fetoprotein-expressing oval cells. Thy-1 protein was consistently present outside the basement membrane surrounding the oval cells. It overlapped frequently with smooth muscle actin staining. A similar cellular localization of the Thy-1 protein was found on human liver specimens with ductular reactions obtained from patients with fulminant liver failure. Furthermore, Thy-1 was expressed by myofibroblasts in experimental liver fibrosis models without oval cell proliferation. We conclude that Thy-1 is not a marker of oval cells but is present on a subpopulation of myofibroblasts/stellate cells. PMID:17884967

  12. A paired comparison of tuberculin skin test results in health care workers using 5 TU and 10 TU tuberculin

    PubMed Central

    Stuart, R.; Bennett, N.; Forbes, A.; Grayson, M

    2000-01-01

    BACKGROUND—Historically, 10 TU has been employed in Australia and the United Kingdom to perform the tuberculin skin test (TST). However, this makes it difficult to compare the rates of TST positivity with other countries such as the USA who use 5 TU. To assess the impact of the dose of tuberculin on the TST a comparison was made of TST responses in health care workers given a TST with both 5 and 10 TU.
METHODS—Two TSTs were performed simultaneously in each health care worker using 5 and 10 TU. Each dose was randomly assigned in a blinded manner to the right or left forearm and read at 48-72 hours by a single nurse who was blinded to the assignment of the 5 and 10 TU doses.
RESULTS—A total of 128 health care workers were enrolled, 119 (93%) of whom had a past history of BCG vaccination. The overall mean difference in paired reaction sizes for the two doses was 1.5 mm with 95% limits of agreement of -3.6 to 6.5 mm.
CONCLUSION—A slightly larger TST reading was seen with 10 TU than with 5 TU. The mean difference of 1.5 mm between the two doses should be considered when comparing rates of TST positivity between countries who use different doses of tuberculin to perform the tuberculin skin test.

 PMID:10899248

  13. Purification of the Thy-1 molecule, a major cell-surface glycoprotein of rat thymocytes.

    PubMed Central

    Letarte-Muirhead, M; Barclay, A N; Williams, A F

    1975-01-01

    The Thy-1-molecule, which was identified by its antigenic activities, has been purified from rat thymocytes. The purification involved preparation of crude membranes and solubilization in deoxycholate, followed by gel filtration and affinity chromatography on antibody or lectin columns. In all cases the purified molecule was a glycoprotein that did not form higher polymers and was not associated with other polypeptide chains. The Thy-1 glycoprotein could be found in two forms, one binding to lentil lectin, the other not. Both forms had the same detectable antigens and were of a similar but not identical size. After sodium dodecyl sulphate-polyacrylamide-gel electrophoresis the apparent molecular weight of Thy-1 binding to lentil lectin was 25 000, whereas that not binding to the lectin was 27 000, with heterogeneity towards forms of apparently higher molecular weight. Images PLATE 4 PLATE 1 PLATE 2 PLATE 3 PMID:56177

  14. 40 CFR 721.600 - 3-Alkyl-2-(2-anilino)vinyl thi-azolinium salt (generic name).

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 30 2010-07-01 2010-07-01 false 3-Alkyl-2-(2-anilino)vinyl thi-azolinium salt (generic name). 721.600 Section 721.600 Protection of Environment ENVIRONMENTAL PROTECTION... New Uses for Specific Chemical Substances § 721.600 3-Alkyl-2-(2-anilino)vinyl thi-azolinium...

  15. 40 CFR 721.600 - 3-Alkyl-2-(2-anilino)vinyl thi-azolinium salt (generic name).

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 32 2013-07-01 2013-07-01 false 3-Alkyl-2-(2-anilino)vinyl thi-azolinium salt (generic name). 721.600 Section 721.600 Protection of Environment ENVIRONMENTAL PROTECTION... New Uses for Specific Chemical Substances § 721.600 3-Alkyl-2-(2-anilino)vinyl thi-azolinium...

  16. 40 CFR 721.600 - 3-Alkyl-2-(2-anilino)vinyl thi-azolinium salt (generic name).

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 32 2012-07-01 2012-07-01 false 3-Alkyl-2-(2-anilino)vinyl thi-azolinium salt (generic name). 721.600 Section 721.600 Protection of Environment ENVIRONMENTAL PROTECTION... New Uses for Specific Chemical Substances § 721.600 3-Alkyl-2-(2-anilino)vinyl thi-azolinium...

  17. 40 CFR 721.600 - 3-Alkyl-2-(2-anilino)vinyl thi-azolinium salt (generic name).

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 31 2014-07-01 2014-07-01 false 3-Alkyl-2-(2-anilino)vinyl thi-azolinium salt (generic name). 721.600 Section 721.600 Protection of Environment ENVIRONMENTAL PROTECTION... New Uses for Specific Chemical Substances § 721.600 3-Alkyl-2-(2-anilino)vinyl thi-azolinium...

  18. 40 CFR 721.600 - 3-Alkyl-2-(2-anilino)vinyl thi-azolinium salt (generic name).

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 31 2011-07-01 2011-07-01 false 3-Alkyl-2-(2-anilino)vinyl thi-azolinium salt (generic name). 721.600 Section 721.600 Protection of Environment ENVIRONMENTAL PROTECTION... New Uses for Specific Chemical Substances § 721.600 3-Alkyl-2-(2-anilino)vinyl thi-azolinium...

  19. Tissue-specific N-glycosylation, site-specific oligosaccharide patterns and lentil lectin recognition of rat Thy-1.

    PubMed Central

    Parekh, R B; Tse, A G; Dwek, R A; Williams, A F; Rademacher, T W

    1987-01-01

    To examine the extent to which protein structure and tissue-type influence glycosylation, we have determined the oligosaccharide structures at each of the three glycosylation sites (Asn-23, 74 and 98) of the cell surface glycoprotein Thy-1 isolated from rat brain and thymus. The results show that there is tissue-specificity of glycosylation and that superimposed on this is a significant degree of site-specificity. On the basis of the site distribution of oligosaccharides, we find that no Thy-1 molecules are in common between the two tissues despite the amino acid sequences being identical. We suggest, therefore, that by controlling N-glycosylation a tissue creates an unique set of glycoforms (same polypeptide but with oligosaccharides that differ either in sequence or disposition). The structures at each of the three sites were also determined for the thymocyte Thy-1 that binds to lentil lectin (Thy-1 L+) and for that which does not (Thy-1 L-). Segregation of intact thymus Thy-1 into two distinct sets of glycoforms by lentil lectin was found to be due to the structures at site 74. Analysis of oligosaccharide structures at the 'passenger' sites (23 and 98) suggests that either Thy-1 L+ and Thy-1 L- molecules are made in different cell-types or that the biosynthesis of oligosaccharides at one site is influenced by the glycosylation at other sites. PMID:2886334

  20. Thy-1 antigen: selective association in lymphoid organs with the vascular basement membrane involved in lymphocyte recirculation.

    PubMed Central

    Ritter, M A; Morris, R J

    1980-01-01

    The cell surface differentiation antigen, Thy-1, was demonstrated by immunofluorescence to be associated with collagen-based connective tissue (mainly basement membrane) around some blood vessels in rat lymphoid organs. This association is highly selective: only certain types of blood vessel within a given lymphoid organ were found to be Thy-1+; and different lymphoid organs (thymus, bone marrow, lymph node and spleen) had characteristic differences in the types of blood vessel that bear Thy-1. In lymph node and spleen the vessels that were Thy-1+ were those involved in lymphocyte recirculation and homing (post-capillary venules and arterioles of white pulp); the possibility that Thy-1 may function in mediating selective adhesion of small lymphocytes to extracellular substrates during recirculation is discussed. Images Figure 2 Figure 3 Figure 1 Figure 4 PMID:6991398

  1. Contribution of Thy1+ NK cells to protective IFN-γ production during Salmonella Typhimurium infections

    PubMed Central

    Kupz, Andreas; Scott, Timothy A.; Belz, Gabrielle T.; Andrews, Daniel M.; Greyer, Marie; Lew, Andrew M.; Brooks, Andrew G.; Smyth, Mark J.; Curtiss, Roy; Bedoui, Sammy; Strugnell, Richard A.

    2013-01-01

    IFN-γ is critical for immunity against infections with intracellular pathogens, such as Salmonella enterica. However, which of the many cell types capable of producing IFN-γ controls Salmonella infections remains unclear. Using a mouse model of systemic Salmonella infection, we observed that only a lack of all lymphocytes or CD90 (Thy1)+ cells, but not the absence of T cells, Retinoic acid-related orphan receptor (ROR)-γt–dependent lymphocytes, (NK)1.1+ cells, natural killer T (NKT), and/or B cells alone, replicated the highly susceptible phenotype of IFN-γ–deficient mice to Salmonella infection. A combination of antibody depletions and adoptive transfer experiments revealed that early protective IFN-γ was provided by Thy1-expressing natural killer (NK) cells and that these cells improved antibacterial immunity through the provision of IFN-γ. Further analysis of NK cells producing IFN-γ in response to Salmonella indicated that less mature NK cells were more efficient at mediating antibacterial effector function than terminally differentiated NK cells. Inspired by recent reports of Thy1+ NK cells contributing to immune memory, we analyzed their role in secondary protection against otherwise lethal WT Salmonella infections. Notably, we observed that a newly generated Salmonella vaccine strain not only conferred superior protection compared with conventional regimens but that this enhanced efficiency of recall immunity was afforded by incorporating CD4−CD8−Thy1+ cells into the secondary response. Taken together, these findings demonstrate that Thy1-expressing NK cells play an important role in antibacterial immunity. PMID:23345426

  2. Astrocytic αVβ3 Integrin Inhibits Neurite Outgrowth and Promotes Retraction of Neuronal Processes by Clustering Thy-1

    PubMed Central

    Herrera-Molina, Rodrigo; Frischknecht, Renato; Maldonado, Horacio; Seidenbecher, Constanze I.; Gundelfinger, Eckart D.; Hetz, Claudio; Aylwin, María de la Luz; Schneider, Pascal; Quest, Andrew F. G.; Leyton, Lisette

    2012-01-01

    Thy-1 is a membrane glycoprotein suggested to stabilize or inhibit growth of neuronal processes. However, its precise function has remained obscure, because its endogenous ligand is unknown. We previously showed that Thy-1 binds directly to αVβ3 integrin in trans eliciting responses in astrocytes. Nonetheless, whether αVβ3 integrin might also serve as a Thy-1-ligand triggering a neuronal response has not been explored. Thus, utilizing primary neurons and a neuron-derived cell line CAD, Thy-1-mediated effects of αVβ3 integrin on growth and retraction of neuronal processes were tested. In astrocyte-neuron co-cultures, endogenous αVβ3 integrin restricted neurite outgrowth. Likewise, αVβ3-Fc was sufficient to suppress neurite extension in Thy-1(+), but not in Thy-1(−) CAD cells. In differentiating primary neurons exposed to αVβ3-Fc, fewer and shorter dendrites were detected. This effect was abolished by cleavage of Thy-1 from the neuronal surface using phosphoinositide-specific phospholipase C (PI-PLC). Moreover, αVβ3-Fc also induced retraction of already extended Thy-1(+)-axon-like neurites in differentiated CAD cells as well as of axonal terminals in differentiated primary neurons. Axonal retraction occurred when redistribution and clustering of Thy-1 molecules in the plasma membrane was induced by αVβ3 integrin. Binding of αVβ3-Fc was detected in Thy-1 clusters during axon retraction of primary neurons. Moreover, αVβ3-Fc-induced Thy-1 clustering correlated in time and space with redistribution and inactivation of Src kinase. Thus, our data indicates that αVβ3 integrin is a ligand for Thy-1 that upon binding not only restricts the growth of neurites, but also induces retraction of already existing processes by inducing Thy-1 clustering. We propose that these events participate in bi-directional astrocyte-neuron communication relevant to axonal repair after neuronal damage. PMID:22479590

  3. EF-Tu dynamics during pre-translocation complex formation: EF-Tu·GDP exits the ribosome via two different pathways

    PubMed Central

    Liu, Wei; Chen, Chunlai; Kavaliauskas, Darius; Knudsen, Charlotte R.; Goldman, Yale E.; Cooperman, Barry S.

    2015-01-01

    The G-protein EF-Tu, which undergoes a major conformational change when EF-Tu·GTP is converted to EF-Tu·GDP, forms part of an aminoacyl(aa)-tRNA·EF-Tu·GTP ternary complex (TC) that accelerates the binding of aa-tRNA to the ribosome during peptide elongation. Such binding, placing a portion of EF-Tu in contact with the GTPase Associated Center (GAC), is followed by GTP hydrolysis and Pi release, and results in formation of a pretranslocation (PRE) complex. Although tRNA movement through the ribosome during PRE complex formation has been extensively studied, comparatively little is known about the dynamics of EF-Tu interaction with either the ribosome or aa-tRNA. Here we examine these dynamics, utilizing ensemble and single molecule assays employing fluorescent labeled derivatives of EF-Tu, tRNA, and the ribosome to measure changes in either FRET efficiency or fluorescence intensity during PRE complex formation. Our results indicate that ribosome-bound EF-Tu separates from the GAC prior to its full separation from aa-tRNA, and suggest that EF-Tu·GDP dissociates from the ribosome by two different pathways. These pathways correspond to either reversible EF-Tu·GDP dissociation from the ribosome prior to the major conformational change in EF-Tu that follows GTP hydrolysis, or irreversible dissociation after or concomitant with this conformational change. PMID:26338772

  4. Applying thiouracil (TU)-tagging for mouse transcriptome analysis

    PubMed Central

    Gay, Leslie; Karfilis, Kate V.; Miller, Michael R.; Doe, Chris Q.; Stankunas, Kryn

    2014-01-01

    Transcriptional profiling is a powerful approach to study mouse development, physiology, and disease models. Here, we describe a protocol for mouse thiouracil-tagging (TU-tagging), a transcriptome analysis technology that includes in vivo covalent labeling, purification, and analysis of cell type-specific RNA. TU-tagging enables 1) the isolation of RNA from a given cell population of a complex tissue, avoiding transcriptional changes induced by cell isolation trauma, and 2) the identification of actively transcribed RNAs and not pre-existing transcripts. Therefore, in contrast to other cell-specific transcriptional profiling methods based on purification of tagged ribosomes or nuclei, TU-tagging provides a direct examination of transcriptional regulation. We describe how to: 1) deliver 4-thiouracil to transgenic mice to thio-label cell lineage-specific transcripts, 2) purify TU-tagged RNA and prepare libraries for Illumina sequencing, and 3) follow a straight-forward bioinformatics workflow to identify cell type-enriched or differentially expressed genes. Tissue containing TU-tagged RNA can be obtained in one day, RNA-Seq libraries generated within two days, and, following sequencing, an initial bioinformatics analysis completed in one additional day. PMID:24457332

  5. Structure of the Mycobacterium tuberculosis Flavin Dependent Thymidylate Synthase (MtbThyX) at 2.0 Å Resolution

    SciTech Connect

    Sampathkumar, Parthasarathy; Turley, Stewart; Ulmer, Jonathan E.; Rhie, Ho Gun; Hopkins Sibley, Carol; Hol, Wim G.J.

    2010-07-20

    A novel flavin-dependent thymidylate synthase was identified recently as an essential gene in many archaebacteria and some pathogenic eubacteria. This enzyme, ThyX, is a potential antibacterial drug target, since humans and most eukaryotes lack the thyX gene and depend upon the conventional thymidylate synthase (TS) for their dTMP requirements. We have cloned and overexpressed the thyX gene (Rv2754c) from Mycobacterium tuberculosis in Escherichia coli. The M. tuberculosis ThyX (MtbThyX) enzyme complements the E. coli {chi}2913 strain that lacks its conventional TS activity. The crystal structure of the homotetrameric MtbThyX was determined in the presence of the cofactor FAD and the substrate analog, 5-bromo-2'-deoxyuridine-5'-monophosphate (BrdUMP). In the active site, which is formed by three monomers, FAD is bound in an extended conformation with the adenosine ring in a deep pocket and BrdUMP in a closed conformation near the isoalloxazine ring. Structure-based mutational studies have revealed a critical role played by residues Lys165 and Arg168 in ThyX activity, possibly by governing access to the carbon atom to be methylated of a totally buried substrate dUMP.

  6. Elongation Factor-Tu (EF-Tu) proteins structural stability and bioinformatics in ancestral gene reconstruction

    NASA Astrophysics Data System (ADS)

    Dehipawala, Sunil; Nguyen, A.; Tremberger, G.; Cheung, E.; Schneider, P.; Lieberman, D.; Holden, T.; Cheung, T.

    2013-09-01

    A paleo-experimental evolution report on elongation factor EF-Tu structural stability results has provided an opportunity to rewind the tape of life using the ancestral protein sequence reconstruction modeling approach; consistent with the book of life dogma in current biology and being an important component in the astrobiology community. Fractal dimension via the Higuchi fractal method and Shannon entropy of the DNA sequence classification could be used in a diagram that serves as a simple summary. Results from biomedical gene research provide examples on the diagram methodology. Comparisons between biomedical genes such as EEF2 (elongation factor 2 human, mouse, etc), WDR85 in epigenetics, HAR1 in human specificity, DLG1 in cognitive skill, and HLA-C in mosquito bite immunology with EF Tu DNA sequences have accounted for the reported circular dichroism thermo-stability data systematically; the results also infer a relatively less volatility geologic time period from 2 to 3 Gyr from adaptation viewpoint. Comparison to Thermotoga maritima MSB8 and Psychrobacter shows that Thermus thermophilus HB8 EF-Tu calibration sequence could be an outlier, consistent with free energy calculation by NUPACK. Diagram methodology allows computer simulation studies and HAR1 shows about 0.5% probability from chimp to human in terms of diagram location, and SNP simulation results such as amoebic meningoencephalitis NAF1 suggest correlation. Extensions to the studies of the translation and transcription elongation factor sequences in Megavirus Chiliensis, Megavirus Lba and Pandoravirus show that the studied Pandoravirus sequence could be an outlier with the highest fractal dimension and lowest entropy, as compared to chicken as a deviant in the DNMT3A DNA methylation gene sequences from zebrafish to human and to the less than one percent probability in computer simulation using the HAR1 0.5% probability as reference. The diagram methodology would be useful in ancestral gene

  7. Domain Organization in Candida glabrata THI6, a Bifunctional Enzyme Required for Thiamin Biosynthesis in Eukaryotes†||‡

    PubMed Central

    Paul, Debamita; Chatterjee, Abhishek; Begley, Tadhg P.; Ealick, Steven E.

    2010-01-01

    THI6 is a bifunctional enzyme found in the thiamin biosynthetic pathway in eukaryotes. The N-terminal domain of THI6 catalyzes the ligation of the thiamin thiazole and pyrimidine moieties to form thiamin phosphate and the C-terminal domain catalyzes the phosphorylation of 4-methyl-5-hydroxyethylthiazole in a salvage pathway. In prokaryotes, thiamin phosphate synthase and 4-methyl-5-hydroxyethylthiazole kinase are separate gene products. Here we report the first crystal structure of a eukaryotic THI6 along with several complexes that characterize the active sites responsible for the two chemical reactions. THI6 from Candida glabrata is a homohexamer in which the six protomers form a cage-like structure. Each protomer is composed of two domains, which are structurally homologous to their monofunctional bacterial counterparts. Two loop regions not found in the bacterial enzymes provide interactions between the two domains. The structures of different protein-ligand complexes define the thiazole and ATP binding sites of the 4-methyl-5-hydroxyethylthiazole kinase domain, and the thiazole phosphate and 4-amino-5-hydroxymethyl-2-methylpyrimidine pyrophosphate binding sites of the thiamin phosphate synthase domain. Our structural studies reveal that the active sites of the two domains are 40 Å apart and are not connected by an obvious channel. Biochemical studies show 4-methyl-5-hydroxyethylthiazole phosphate is a substrate for THI6; however, adenosine diphospho-5-β-ethyl-4-methylthiazole-2-carboxylic acid, the product of THI4, is not a substrate for THI6. This suggests that unidentified enzyme is necessary to produce the substrate for THI6 from the THI4 product. PMID:20968298

  8. THY-1 Cell Surface Antigen (CD90) Has an Important Role in the Initial Stage of Human Cytomegalovirus Infection

    PubMed Central

    Li, Qingxue; Wilkie, Adrian R.; Weller, Melodie; Liu, Xueqiao; Cohen, Jeffrey I.

    2015-01-01

    Human cytomegalovirus (HCMV) infects about 50% of the US population, is the leading infectious cause of birth defects, and is considered the most important infectious agent in transplant recipients. The virus infects many cell types in vivo and in vitro. While previous studies have identified several cellular proteins that may function at early steps of infection in a cell type dependent manner, the mechanism of virus entry is still poorly understood. Using a computational biology approach, correlating gene expression with virus infectivity in 54 cell lines, we identified THY-1 as a putative host determinant for HCMV infection in these cells. With a series of loss-of-function, gain-of-function and protein-protein interaction analyses, we found that THY-1 mediates HCMV infection at the entry step and is important for infection that occurs at a low m.o.i. THY-1 antibody that bound to the cell surface blocked HCMV during the initial 60 minutes of infection in a dose-dependent manner. Down-regulation of THY-1 with siRNA impaired infectivity occurred during the initial 60 minutes of inoculation. Both THY-1 antibody and siRNA inhibited HCMV-induced activation of the PI3-K/Akt pathway required for entry. Soluble THY-1 protein blocked HCMV infection during, but not after, virus internalization. Expression of exogenous THY-1 enhanced entry in cells expressing low levels of the protein. THY-1 interacted with HCMV gB and gH and may form a complex important for entry. However, since gB and gH have previously been shown to interact, it is uncertain if THY-1 directly binds to both of these proteins. Prior observations that THY-1 (a) interacts with αVβ3 integrin and recruits paxillin (implicated in HCMV entry), (b) regulates leukocyte extravasation (critical for HCMV viremia), and (c) is expressed on many cells targeted for HCMV infection including epithelial and endothelial cells, fibroblast, and CD34+/CD38- stem cells, all support a role for THY-1 as an HCMV entry mediator in

  9. THY-1 Cell Surface Antigen (CD90) Has an Important Role in the Initial Stage of Human Cytomegalovirus Infection.

    PubMed

    Li, Qingxue; Wilkie, Adrian R; Weller, Melodie; Liu, Xueqiao; Cohen, Jeffrey I

    2015-07-01

    Human cytomegalovirus (HCMV) infects about 50% of the US population, is the leading infectious cause of birth defects, and is considered the most important infectious agent in transplant recipients. The virus infects many cell types in vivo and in vitro. While previous studies have identified several cellular proteins that may function at early steps of infection in a cell type dependent manner, the mechanism of virus entry is still poorly understood. Using a computational biology approach, correlating gene expression with virus infectivity in 54 cell lines, we identified THY-1 as a putative host determinant for HCMV infection in these cells. With a series of loss-of-function, gain-of-function and protein-protein interaction analyses, we found that THY-1 mediates HCMV infection at the entry step and is important for infection that occurs at a low m.o.i. THY-1 antibody that bound to the cell surface blocked HCMV during the initial 60 minutes of infection in a dose-dependent manner. Down-regulation of THY-1 with siRNA impaired infectivity occurred during the initial 60 minutes of inoculation. Both THY-1 antibody and siRNA inhibited HCMV-induced activation of the PI3-K/Akt pathway required for entry. Soluble THY-1 protein blocked HCMV infection during, but not after, virus internalization. Expression of exogenous THY-1 enhanced entry in cells expressing low levels of the protein. THY-1 interacted with HCMV gB and gH and may form a complex important for entry. However, since gB and gH have previously been shown to interact, it is uncertain if THY-1 directly binds to both of these proteins. Prior observations that THY-1 (a) interacts with αVβ3 integrin and recruits paxillin (implicated in HCMV entry), (b) regulates leukocyte extravasation (critical for HCMV viremia), and (c) is expressed on many cells targeted for HCMV infection including epithelial and endothelial cells, fibroblast, and CD34+/CD38- stem cells, all support a role for THY-1 as an HCMV entry mediator in

  10. An unprecedented mechanism of nucleotide methylation in organisms containing thyX.

    PubMed

    Mishanina, Tatiana V; Yu, Liping; Karunaratne, Kalani; Mondal, Dibyendu; Corcoran, John M; Choi, Michael A; Kohen, Amnon

    2016-01-29

    In several human pathogens, thyX-encoded flavin-dependent thymidylate synthase (FDTS) catalyzes the last step in the biosynthesis of thymidylate, one of the four DNA nucleotides. ThyX is absent in humans, rendering FDTS an attractive antibiotic target; however, the lack of mechanistic understanding prohibits mechanism-based drug design. Here, we report trapping and characterization of two consecutive intermediates, which together with previous crystal structures indicate that the enzyme's reduced flavin relays a methylene from the folate carrier to the nucleotide acceptor. Furthermore, these results corroborate an unprecedented activation of the nucleotide that involves no covalent modification but only electrostatic polarization by the enzyme's active site. These findings indicate a mechanism that is very different from thymidylate biosynthesis in humans, underscoring the promise of FDTS as an antibiotic target. PMID:26823429

  11. Two populations of Thy1-positive mesenchymal cells regulate in vitro maturation of hepatic progenitor cells.

    PubMed

    Kamo, Naoko; Yasuchika, Kentaro; Fujii, Hideaki; Hoppo, Toshitaka; Machimoto, Takafumi; Ishii, Takamichi; Fujita, Naoya; Tsuruo, Takashi; Yamashita, Jun K; Kubo, Hajime; Ikai, Iwao

    2007-02-01

    We previously reported that the in vitro maturation of CD49f(+)Thy1(-)CD45(-) (CD49f positive) fetal hepatic progenitor cells (HPCs) is supported by Thy1-positive mesenchymal cells derived from the fetal liver. These mesenchymal cell preparations contain two populations, one of a cuboidal shape and the other spindle shaped in morphology. In this study, we determined that the mucin-type transmembrane glycoprotein gp38 could distinguish cuboidal cells from spindle cells by immunocytochemistry. RT-PCR analysis revealed differences between isolated CD49f(+/-)Thy1(+)gp38(+)CD45(-) (gp38 positive) cells and CD49f(+/-)Thy1(+)gp38(-)CD45(-) (gp38 negative) cells, whereas both cells expressed mesenchymal cell markers. The coculture with gp38-positive cells promoted the maturation of CD49f-positive HPCs, which was estimated by positivity for periodic acid-Schiff (PAS) staining, whereas the coculture with gp38-negative cells maintained CD49f-positive HPCs negative for PAS staining. The expression of mature hepatocyte markers, such as tyrosine aminotransferase, tryptophan-2,3-dioxygenase, and glucose-6-phosphatase, were upregulated on HPCs by coculture with gp38-positive cells. Furthermore, transmission electron microscopy revealed the acquisition of mature hepatocyte features by HPCs cocultured with gp38-positive cells. This effect on maturation of HPCs was inhibited by the addition of conditioned medium derived from gp38-negative cells. By contrast, the upregulation of bromodeoxyuridine incorporation by HPCs demonstrated the proliferative effect of coculture with gp38-negative cells. In conclusion, these results suggest that in vitro maturation of HPCs promoted by gp38-positive cells may be opposed by an inhibitory effect of gp38-negative cells, which likely maintain the immature, proliferative state of HPCs. PMID:16990447

  12. Predictive modeling targets thymidylate synthase ThyX in Mycobacterium tuberculosis.

    PubMed

    Djaout, Kamel; Singh, Vinayak; Boum, Yap; Katawera, Victoria; Becker, Hubert F; Bush, Natassja G; Hearnshaw, Stephen J; Pritchard, Jennifer E; Bourbon, Pauline; Madrid, Peter B; Maxwell, Anthony; Mizrahi, Valerie; Myllykallio, Hannu; Ekins, Sean

    2016-01-01

    There is an urgent need to identify new treatments for tuberculosis (TB), a major infectious disease caused by Mycobacterium tuberculosis (Mtb), which results in 1.5 million deaths each year. We have targeted two essential enzymes in this organism that are promising for antibacterial therapy and reported to be inhibited by naphthoquinones. ThyX is an essential thymidylate synthase that is mechanistically and structurally unrelated to the human enzyme. DNA gyrase is a DNA topoisomerase present in bacteria and plants but not animals. The current study set out to understand the structure-activity relationships of these targets in Mtb using a combination of cheminformatics and in vitro screening. Here, we report the identification of new Mtb ThyX inhibitors, 2-chloro-3-(4-methanesulfonylpiperazin-1-yl)-1,4-dihydronaphthalene-1,4-dione) and idebenone, which show modest whole-cell activity and appear to act, at least in part, by targeting ThyX in Mtb. PMID:27283217

  13. Predictive modeling targets thymidylate synthase ThyX in Mycobacterium tuberculosis

    PubMed Central

    Djaout, Kamel; Singh, Vinayak; Boum, Yap; Katawera, Victoria; Becker, Hubert F.; Bush, Natassja G.; Hearnshaw, Stephen J.; Pritchard, Jennifer E.; Bourbon, Pauline; Madrid, Peter B.; Maxwell, Anthony; Mizrahi, Valerie; Myllykallio, Hannu; Ekins, Sean

    2016-01-01

    There is an urgent need to identify new treatments for tuberculosis (TB), a major infectious disease caused by Mycobacterium tuberculosis (Mtb), which results in 1.5 million deaths each year. We have targeted two essential enzymes in this organism that are promising for antibacterial therapy and reported to be inhibited by naphthoquinones. ThyX is an essential thymidylate synthase that is mechanistically and structurally unrelated to the human enzyme. DNA gyrase is a DNA topoisomerase present in bacteria and plants but not animals. The current study set out to understand the structure-activity relationships of these targets in Mtb using a combination of cheminformatics and in vitro screening. Here, we report the identification of new Mtb ThyX inhibitors, 2-chloro-3-(4-methanesulfonylpiperazin-1-yl)-1,4-dihydronaphthalene-1,4-dione) and idebenone, which show modest whole-cell activity and appear to act, at least in part, by targeting ThyX in Mtb. PMID:27283217

  14. Bacteriophage Resistance of a ΔthyA Mutant of Lactococcus lactis Blocked in DNA Replication

    PubMed Central

    Pedersen, Martin B.; Jensen, Peter R.; Janzen, Thomas; Nilsson, Dan

    2002-01-01

    The thyA gene, which encodes thymidylate synthase (TS), of Lactococcus lactis CHCC373 was sequenced, including the upstream and downstream regions. We then deleted part of thyA by gene replacement. The resulting strain, MBP71 ΔthyA, was devoid of TS activity, and in media without thymidine, such as milk, there was no detectable dTTP pool in the cells. Hence, DNA replication was abolished, and acidification by MBP71 was completely unaffected by the presence of nine different phages tested at a multiplicity of infection (MOI) of 0.1. Nonreplicating MBP71 must be inoculated at a higher level than CHCC373 to achieve a certain pH within a specified time. For a pH of 5.2 to be reached in 6 h, the inoculation level of MBP71 must be 17-fold higher than for CHCC373. However, by adding a limiting amount of thymidine this could be lowered to just 5-fold the normal amount, while acidification was unaffected with MBP71 up to an MOI of 0.01. It was found that nonreplicating MBP71 produced largely the same products as CHCC373, though the acetaldehyde production of the former was higher. PMID:12039762

  15. Conformational changes in the expression domain of the Escherichia coli thiM riboswitch

    PubMed Central

    Rentmeister, Andrea; Mayer, Günter; Kuhn, Nicole; Famulok, Michael

    2007-01-01

    The thiM riboswitch contains an aptamer domain that adaptively binds the coenzyme thiamine pyrophosphate (TPP). The binding of TPP to the aptamer domain induces structural rearrangements that are relayed to a second domain, the so-called expression domain, thereby interfering with gene expression. The recently solved crystal structures of the aptamer domains of the thiM riboswitches in complex with TPP revealed how TPP stabilizes secondary and tertiary structures in the RNA ligand complex. To understand the global modes of reorganization between the two domains upon metabolite binding the structure of the entire riboswitch in presence and absence of TPP needs to be determined. Here we report the secondary structure of the entire thiM riboswitch from Escherichia coli in its TPP-free form and its transition into the TPP-bound variant, thereby depicting domains of the riboswitch that serve as communication links between the aptamer and the expression domain. Furthermore, structural probing provides an explanation for the lack of genetic control exerted by a riboswitch variant with mutations in the expression domain that still binds TPP. PMID:17517779

  16. A Grammar of Bao'an Tu, a Mongolic Language of Northwest China

    ERIC Educational Resources Information Center

    Fried, Robert Wayne

    2010-01-01

    The present study is a grammatical overview of the Bao'an Tu language (one of the varieties included in the designation "ISO6393-3:PEH", also known as "Tongren Monguor", "Southwestern Monguor", or "Tongren Tu"). Bao'an Tu is spoken by approximately 4,000 people who live on the Qinghai-Tibetan Plateau in Tongren County, Huangnan Prefecture, Qinghai…

  17. The thiG Gene Is Required for Full Virulence of Xanthomonas oryzae pv. oryzae by Preventing Cell Aggregation

    PubMed Central

    Yu, Xiaoyue; Liang, Xiaoyu; Liu, Kexue; Dong, Wenxia; Wang, Jianxin; Zhou, Ming-guo

    2015-01-01

    Bacterial blight of rice is an important serious bacterial diseases of rice in many rice-growing regions, caused by Xanthomonas oryzae pv. oryzae (Xoo). The thiG gene from Xoo strain ZJ173, which is involved with thiazole moiety production in the thiamine biosynthesis pathway, is highly conserved among the members of Xanthomonas. The thiG deletion mutant displayed impaired virulence and growth in thiamine-free medium but maintained its normal growth rate in the rice tissues, indicating that the thiG gene is involved in Xoo virulence. Compared to the wild type strain, the formation of cell-cell aggregates was affected in thiG deletion mutants. Although biofilm formation was promoted, motility and migration in rice leaves were repressed in the thiG mutants, and therefore limited the expansion of pathogen infection in rice. Quorum sensing and extracellular substance are two key factors that contribute to the formation of cell-cell aggregates. Our study found that in the thiG mutant the expression of two genes, rpfC and rpfG, which form a two-component regulatory signal system involved in the regulation of biofilm formation by a second messenger cyclic di-GMP is down-regulated. In addition, our study showed that xanthan production was not affected but the expression of some genes associated with xanthan biosynthesis, like gumD, gumE, gumH and gumM, were up-regulated in thiG mutants. Taken together, these findings are the first to demonstrate the role of the thiazole biosynthsis gene, thiG, in virulence and the formation of aggregates in Xanthomonas oryzae pv. oryzae. PMID:26222282

  18. "The Et Tu Brute Complex" Compulsive Self Betrayal

    ERIC Educational Resources Information Center

    Antus, Robert Lawrence

    2006-01-01

    In this article, the author discusses "The Et Tu Brute Complex." More specifically, this phenomenon occurs when a person, instead of supporting and befriending himself, orally condemns himself in front of other people and becomes his own worst enemy. This is a form of compulsive self-hatred. Most often, the victim of this complex is unaware of the…

  19. The Impending Demise of "tu" in Montevideo, Uruguay

    ERIC Educational Resources Information Center

    Weyers, Joseph R.

    2009-01-01

    Since the early 1960s, the "tuteo" (T-T), and particularly the use of tonic "tu" with the "vos" verbal morphology (T-V) has been accepted as typical of the speech of Montevideo, Uruguay, a region generally classified as "voseante". This paper reports on the results of 117 rapid anonymous interviews conducted in Montevideo to document the status of…

  20. Lymphopenic effects on mice of a component of ammonia caramel, 2-acetyl-4(5)-tetrahydroxybutylimidazole (THI).

    PubMed

    Iscaro, A; Mackay, I R; O'Brien, C

    1988-01-01

    A derivative of ammonia caramel colour (AC) is known to induce a selective lymphopenia in rats. Accordingly, the haematological effects were studied in mice of oral administration in drinking water of 2-acetyl-4(5)-tetrahydroxybutylimidazole (THI), the component of AC responsible for lymphopenia. Initially five groups of BALB/c mice (five mice per group) were given doses of THI ranging from 0 to 200 parts/10(6) and bled weekly. Doses of THI from 5 to 100 parts/10(6) had no effect on circulating leucocytes over 6 weeks, but lymphopenia occurred with 200 parts/10(6). An increase in the concentration of THI to 400 parts/10(6) in the group on the lowest dose resulted in lymphopenia. An increase in dosage in two groups of mice, to 1000 and 2000 parts/10(6), resulted in marked lymphopenia. The number of neutrophils, eosinophils and monocytes remained unchanged throughout the experiment. Measurement of the proportions of CD4(L3T4)+ and CD8(Ly2)+ lymphocytes in lymph nodes from mice on high doses of THI did not show a selective depression of either subset, although both were increased relative to non-T cells. THI causes a selective lymphopenia in mice, as in rats, but at relatively higher doses, and merits investigation in mice as an experimental treatment for states of lymphocyte excess or overactivity. PMID:3147240

  1. Expression, purification, and characterization of avian Thy-1 from Lec1 mammalian and Tn5 insect cells.

    PubMed

    Mehndiratta, Promod; Walton, Wendy J; Hare, Joan T; Pulido, Silvia; Parthasarathy, Gopalakrishnan; Emmett, Mark R; Marshall, Alan G; Logan, Timothy M

    2004-02-01

    Structural studies of asparagine-linked glycoproteins are complicated by the oligosaccharide heterogeneity inherent to individual glycosylation sites. Herein, we report the cloning of a novel isoform of avian Thy-1 and the subsequent expression, purification, and characterization of a soluble form of Thy-1 from Lec1 mammalian and Tn5 insect cells. The novel isoform of Thy-1 differs from the previously reported chicken isoform by eight amino acid residues, but these changes do not alter the secondary structure content, the disulfide bond pattern, or the sites of glycosylation. The disulfide linkage pattern and glycoform distribution on each N-glycosylation site of recombinant chicken Thy-1 from both cell lines were determined by a combination of amino-terminal sequencing and mass spectrometry. The mass spectral data showed that the amino-terminal glutamine was modified to pyroglutamate. Recombinant Thy-1 from Lec1 cells contained (GlcNAc)(2)(Man)(5) on asparagine 60, whereas the oligosaccharides on asparagine 23 and 100 contained approximately 80% (GlcNAc)(2)(Man)(4) and approximately 20% (GlcNAc)(2)(Man)(5). The glycoforms on Thy-1 expressed in Tn5 cells were more heterogeneous, with the oligosaccharides ranging over (GlcNAc)(2)(Fuc)(0-2)(Man)(2-3) on each site. The ability to generate recombinant glycoproteins with restricted carbohydrate heterogeneity is the first step toward the systematic study of structure-function relationships in intact glycoproteins. PMID:14711516

  2. Modification of amino groups in EF-Tu.GTP and the ternary complex EF-Tu.GTP.valyl-tRNAVal.

    PubMed

    Antonsson, B; Leberman, R

    1984-06-15

    In an attempt to describe the binding region of EF-Tu . GTP for aminoacyl-tRNA, the epsilon-amino groups of the lysine residues of the protein molecule in the GTP and ternary complexes were modified with ethyl acetimidate. Using [14C]ethyl acetimidate, tryptic digestion, fractionation of peptides by high-performance liquid chromatography, and amino acid analysis, all reactive lysine residues could be unambiguously identified. 19 of the 23 lysine residues of EF-Tu were labelled under conditions for ternary complex stability. Of these only 8 showed differences in reactivity between free and complexed EF-Tu . GTP. In the ternary complex lysine residues 208 and 390 [Jones, M. D., Petersen, T. E., Nielsen, K. M., Magnusson, S., Sotterup-Jensen, L., Gausing, K. and Clark, B. F. C. (1980) Eur. J. Biochem. 108, 507-526] showed an increase in reactivity (60% and 30% respectively) and residues 2, 4, 237, 248, 263, and 282 showed a decrease in reactivity (between 85% and 37%) compared to the values observed with EF-Tu . GTP. The greatest changes in reactivity were observed for lysine residues 2, 4 and 263. These data can be combined with the available structural information to identify possible areas of contact between the protein and nucleic acid moieties in the ternary complex. PMID:6430701

  3. Renal tubular angiogenic dysregulation in anti-Thy1.1 glomerulonephritis.

    PubMed

    Cina, Davide P; Xu, Hui; Liu, Limin; Farkas, Laszlo; Farkas, Daniela; Kolb, Martin; Margetts, Peter J

    2011-02-01

    Peritubular vascular changes and hypoxia after glomerular injury may explain subsequent tubulointerstitial injury and fibrosis. Several studies suggested that the expected tubulointerstitial angiogenic response is actively suppressed in this setting. The mechanism of this aberrant response has not been clearly identified. We used a common model of glomerular injury in rats to assess vascular changes and to identify potential factors associated with this aberrant response. Anti-Thy1.1 antibody administration (1 or 4 weekly doses) led to a dose-dependent renal damage characterized by elevated urea and tubulointerstitial fibrosis as assessed by Picro-Sirius Red staining. We quantified peritubular capillaries using CD31 and CD34 immunohistochemistry and showed that tubular angiogenic dysregulation was associated with peritubular capillary rarefaction. Using laser capture microdissection, we demonstrated an early induction of fibrogenic and angiogenic factors in the glomeruli and a subsequent dysregulated angiogenic response in the tubulointerstitial compartment. Proximal tubules of anti-Thy1.1-treated animals had increased pigment epithelial-derived factor (PEDF) expression by immunohistochemistry. Protein taken by laser capture microdissection also showed that PEDF was upregulated. Temporally associated with PEDF expression was a transient downregulation of tubular hypoxia-inducible factor (HIF)1α. In a human proximal tubular cell culture, we show that PEDF downregulates HIF1α protein and gene expression in cells exposed to 1% oxygen. In anti-Thy1.1 glomerulonephritis, there is aberrent tubular angiogenesis associated with glomerular injury and tubulointersititial fibrosis. We showed that PEDF may be involved by downregulating HIF1α. Further work is needed to elucidate the mechanism of PEDF upregulation and action in the tubules. PMID:21048020

  4. IL-17 Expression in the Time Course of Acute Anti-Thy1 Glomerulonephritis

    PubMed Central

    Loof, Tanja; Krämer, Stephanie; Gaedeke, Jens; Neumayer, Hans-Hellmut; Peters, Harm

    2016-01-01

    Background Interleukin-17 (IL-17) is a new pro-inflammatory cytokine involved in immune response and inflammatory disease. The main source of IL-17 is a subset of CD4+ T-helper cells, but is also secreted by non-immune cells. The present study analyzes expression of IL-17 in the time course of acute anti-thy1 glomerulonephritis and the role of IL-17 as a potential link between inflammation and fibrosis. Methods Anti-thy1 glomerulonephritis was induced into male Wistar rats by OX-7 antibody injection. After that, samples were taken on days 1, 5, 10 (matrix expansion phase), 15 and 20 (resolution phase). PBS-injected animals served as controls. Proteinuria and histological matrixes score served as the main markers for disease severity. In in vitro experiments, NRK-52E cells were used. For cytokine expressions, mRNA and protein levels were analyzed by utilizing RT-PCR, in situ hybridization and immunofluorescence. Results Highest IL-17 mRNA-expression (6.50-fold vs. con; p<0.05) was found on day 5 after induction of anti-thy1 glomerulonephritis along the maximum levels of proteinuria (113 ± 13 mg/d; p<0.001), histological glomerular-matrix accumulation (82%; p<0.001) and TGF-β1 (2.2-fold; p<0.05), IL-6 mRNA expression (36-fold; p<0.05). IL-17 protein expression co-localized with the endothelial cell marker PECAM in immunofluorescence. In NRK-52E cells, co-administration of TGF-β1 and IL-6 synergistically up-regulated IL-17 mRNA 4986-fold (p<0.001). Conclusions The pro-inflammatory cytokine IL-17 is up-regulated in endothelial cells during the time course of acute anti-thy1 glomerulonephritis. In vitro, NRK-52E cells secrete IL-17 under pro-fibrotic and pro-inflammatory conditions. PMID:27243813

  5. Transcriptional unit of the murine Thy-1 gene: different distribution of transcription initiation sites in brain.

    PubMed Central

    Spanopoulou, E; Giguere, V; Grosveld, F

    1988-01-01

    Structural analysis of the mouse Thy-1.2 gene has shown that the major promoter of the gene is characterized by a tissue-specific DNase I-hypersensitive site and is located within a methylation-free island. The gene is regulated at the transcriptional level, and steady-state mRNA analysis reveals that the previously reported exon Ib contributes at most 5% of the total mRNA. The major promoter uses several transcription initiation sites within a region of 100 base pairs. The frequency of usage of these sites in brain is markedly different from that in other tissues. Images PMID:2906111

  6. Identification and cloning of two immunogenic C. perfringens proteins, elongation factor Tu (EF-Tu) and pyruvate:ferredoxin oxidoreductase (PFO) of Clostridium perfringens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Clostridium related poultry diseases such as necrotic enteritis (NE) and gangrenous dermatitis (GD) cause substantial economic losses on a global scale. Two antigenic C. perfringens proteins, elongation factor Tu (EF-Tu) and pyruvate:ferredoxin oxidoreductase (PFO), were identified by reaction with...

  7. Kirromycin-induced modifications facilitate the separation of EF-Tu species and reveal intermolecular interactions.

    PubMed

    Anborgh, P H; Swart, G W; Parmeggiani, A

    1991-11-01

    A simplified method for the separation of a kirromycin-sensitive tufB-encoded elongation factor Tu (EF-TuBs) from a kirromycin-resistant tufA product (EF-TuAr) was obtained by exploiting the specific increase of negative [corrected] charges induced by the antibiotic, resulting in a retarded elution of kirromycin-bound EF-TuBs on ionic chromatography. The kirromycin-free EF-TuBs is active in poly(Phe) synthesis and shows similar properties to EF-TuAsBs. As expected for these two distinct species, the dissociation of the EF-TuArBs.GTP complex in the presence of kirromycin shows a biphasic curve; in contrast, a monophasic GTP dissociation rate was found for a combination of two mutated EF-Tu species, EF-TuArBo, revealing the existence of intermolecular interactions. These observations prove for the first time the existence of cooperative phenomena between EF-Tu species in vitro, as suggested earlier by in vivo experiments. PMID:1959611

  8. Prevention of spontaneous and cyclophosphamide-induced diabetes in non-obese diabetic (NOD) mice with oral 2-acetyl-4-tetrahydroxybutylimidazole (THI), a component of caramel colouring III.

    PubMed Central

    Mandel, T E; Koulmanda, M; Mackay, I R

    1992-01-01

    The effect of oral administration of THI, a compound present in ammonia caramel food colouring, was studied in spontaneous and induced murine diabetes mellitus. Continuous administration of THI at 400 ppm in drinking water reduced the prevalence of spontaneous diabetes in female NOD/Lt mice from 63% in untreated controls to 8% in treated animals. Since cyclophosphamide (CP) accelerates and intensifies diabetes in NOD mice, we also studied the effect of THI in this model. Diabetes incidence was reduced from 100% in mice given only CP to 13-14% in mice given THI either concurrently or from 14 days previously. Histologically, THI greatly reduced the severity of insulitis. As measured by flow cytometry, all THI-treated mice had a 60-80% reduction in splenic CD4+ and CD8+ T cells. THI-treated mice showed no untoward effects and specifically no weight loss, or pathological changes in their livers, kidneys or lungs. However, there was moderate atrophy of the thymus cortex. THI is a small imidazole-containing compound with structural similarity to histamine and urocanic acid, both known to have immunosuppressive properties. It is a widely used food additive with no known long-term toxic effects at low dosage. Thus, THI could be a useful immunosuppressive agent. PMID:1606724

  9. Tu-144LL SST Flying Laboratory Side View of Nose, with a TU-144D on Ramp

    NASA Technical Reports Server (NTRS)

    1998-01-01

    A Tupolev Tu-144D supersonic jetliner is framed by the drooped nose and forward fuselage of the Tu-144LL supersonic flying laboratory at the Zhukovsky Air Development Center near Moscow, Russia, in 1998. NASA teamed with American and Russian aerospace industries for an extended period in a joint international research program featuring the Russian-built Tu-144LL supersonic aircraft. The object of the program was to develop technologies for a proposed future second-generation supersonic airliner to be developed in the 21st Century. The aircraft's initial flight phase began in June 1996 and concluded in February 1998 after 19 research flights. A shorter follow-on program involving seven flights began in September 1998 and concluded in April 1999. All flights were conducted in Russia from Tupolev's facility at the Zhukovsky Air Development Center near Moscow. The centerpiece of the research program was the Tu 144LL, a first-generation Russian supersonic jetliner that was modified by its developer/builder, Tupolev ANTK (aviatsionnyy nauchno-tekhnicheskiy kompleks-roughly, aviation technical complex), into a flying laboratory for supersonic research. Using the Tu-144LL to conduct flight research experiments, researchers compared full-scale supersonic aircraft flight data with results from models in wind tunnels, computer-aided techniques, and other flight tests. The experiments provided unique aerodynamic, structures, acoustics, and operating environment data on supersonic passenger aircraft. Data collected from the research program was being used to develop the technology base for a proposed future American-built supersonic jetliner. Although actual development of such an advanced supersonic transport (SST) is currently on hold, commercial aviation experts estimate that a market for up to 500 such aircraft could develop by the third decade of the 21st Century. The Tu-144LL used in the NASA-sponsored research program was a 'D' model with different engines than were used

  10. Imaging Neural Activity Using Thy1-GCaMP Transgenic mice

    PubMed Central

    Chen, Qian; Cichon, Joseph; Wang, Wenting; Qiu, Li; Lee, Seok-Jin R.; Campbell, Nolan R.; DeStefino, Nicholas; Goard, Michael J.; Fu, Zhanyan; Yasuda, Ryohei; Looger, Loren L.; Arenkiel, Benjamin R.; Gan, Wen-Biao; Feng, Guoping

    2014-01-01

    Summary The ability to chronically monitor neuronal activity in the living brain is essential for understanding the organization and function of the nervous system. The genetically encoded green fluorescent protein based calcium sensor GCaMP provides a powerful tool for detecting calcium transients in neuronal somata, processes, and synapses that are triggered by neuronal activities. Here we report the generation and characterization of transgenic mice that express improved GCaMPs in various neuronal subpopulations under the control of the Thy1 promoter. In vitro and in vivo studies show that calcium transients induced by spontaneous and stimulus-evoked neuronal activities can be readily detected at the level of individual cells and synapses in acute brain slices, as well as chronically in awake behaving animals. These GCaMP transgenic mice allow investigation of activity patterns in defined neuronal populations in the living brain, and will greatly facilitate dissecting complex structural and functional relationships of neural networks. PMID:23083733

  11. Mannose-6-phosphate facilitates early peripheral nerve regeneration in thy-1-YFP-H mice.

    PubMed

    Harding, A J; Christmas, C R; Ferguson, M W J; Loescher, A R; Robinson, P P; Boissonade, F M

    2014-10-24

    The formation of scar tissue following nerve injury has been shown to adversely affect nerve regeneration and evidence suggests that mannose-6-phosphate (M6P), a potential scar reducing agent that affects transforming growth factor (TGF)-β activation, may enhance nerve regeneration. In this study we utilized thy-1-YFP-H mice - a transgenic strain expressing yellow fluorescent protein (YFP) within a subset of axons - to enable visual analysis of axons regenerating through a nerve graft. Using this strain of mouse we have developed analysis techniques to visualize and quantify regeneration of individual axons across the injury site following the application of either M6P or vehicle to the site of nerve injury. No significant differences were found in the proportion of axons regenerating through the graft between M6P- and vehicle-treated grafts at any point along the graft length. Maximal sprouting occurred at 1.0mm from the proximal graft ending in both groups. The maximum change in sprouting levels for both treatment groups occurred between the graft start and 0.5-mm interval for both treatment groups. The difference between repair groups was significant at this point with a greater increase seen in the vehicle group than the M6P group. The average length of axons regenerating across the initial graft entry was significantly shorter in M6P- than in vehicle-treated grafts, indicating that they encountered less impedance. Application of M6P appears to reduce the disruption of regenerating axons and may therefore facilitate quicker recovery; this is likely to result from altered scar tissue formation in M6P grafts in the early stages of recovery. This study also establishes the usefulness of our methods of analysis using the thy-1-YFP-H mouse strain to visualize and quantify regeneration at the level of the individual axon. PMID:25173153

  12. thyA as a Selection Marker in Construction of Food-Grade Host-Vector and Integration Systems for Streptococcus thermophilus

    PubMed Central

    Sasaki, Yasuko; Ito, Yoshiyuki; Sasaki, Takashi

    2004-01-01

    We constructed food-grade host-vector and integration systems for Streptococcus thermophilus by using a thymidylate synthase gene (thyA) as the selection marker. Two thyA genes, thyASt and thyALb, were cloned from S. thermophilus and Lactobacillus delbrueckii subsp. bulgaricus, respectively. Thymidine-requiring mutants of S. thermophilus were obtained after successive cultures in the presence of trimethoprim, and one of them, TM1-1, was used as the host. Food-grade vectors were constructed by using either thyASt or thyALb as the selection marker. Transformants of TM1-1 created by using these vectors were selected for thymidine autotrophy as efficiently as for erythromycin resistance. By using the host-vector system developed in this way, a foreign amylase gene (amyA) was expressed in TM1-1 and was also integrated into the chromosome by use of a temperature-sensitive integration vector constructed with thyALb as the selection marker via a double-crossover event. The results obtained show that thyA is an efficient and safe selection marker for S. thermophilus that is suitable for food applications. PMID:15006818

  13. Tu-144LL SST Flying Laboratory in Flight

    NASA Technical Reports Server (NTRS)

    1998-01-01

    The delta wing of the Tupolev Tu-144LL supersonic flying laboratory is evident in this view from underneath the aircraft during a 1998 test flight at the Zhukovsky Air Development Center near Moscow, Russia. NASA teamed with American and Russian aerospace industries for an extended period in a joint international research program featuring the Russian-built Tu-144LL supersonic aircraft. The object of the program was to develop technologies for a proposed future second-generation supersonic airliner to be developed in the 21st Century. The aircraft's initial flight phase began in June 1996 and concluded in February 1998 after 19 research flights. A shorter follow-on program involving seven flights began in September 1998 and concluded in April 1999. All flights were conducted in Russia from Tupolev's facility at the Zhukovsky Air Development Center near Moscow. The centerpiece of the research program was the Tu 144LL, a first-generation Russian supersonic jetliner that was modified by its developer/builder, Tupolev ANTK (aviatsionnyy nauchno-tekhnicheskiy kompleks-roughly, aviation technical complex), into a flying laboratory for supersonic research. Using the Tu-144LL to conduct flight research experiments, researchers compared full-scale supersonic aircraft flight data with results from models in wind tunnels, computer-aided techniques, and other flight tests. The experiments provided unique aerodynamic, structures, acoustics, and operating environment data on supersonic passenger aircraft. Data collected from the research program was being used to develop the technology base for a proposed future American-built supersonic jetliner. Although actual development of such an advanced supersonic transport (SST) is currently on hold, commercial aviation experts estimate that a market for up to 500 such aircraft could develop by the third decade of the 21st Century. The Tu-144LL used in the NASA-sponsored research program was a 'D' model with different engines than were

  14. Tu-144LL SST Flying Laboratory in Flight

    NASA Technical Reports Server (NTRS)

    1998-01-01

    The Tupolev Tu-144LL supersonic flying laboratory shows off its sleek lines in a low-level pass over the Zhukovsky Air Development Center near Moscow, Russia, on a 1998 research flight. NASA teamed with American and Russian aerospace industries for an extended period in a joint international research program featuring the Russian-built Tu-144LL supersonic aircraft. The object of the program was to develop technologies for a proposed future second-generation supersonic airliner to be developed in the 21st Century. The aircraft's initial flight phase began in June 1996 and concluded in February 1998 after 19 research flights. A shorter follow-on program involving seven flights began in September 1998 and concluded in April 1999. All flights were conducted in Russia from Tupolev's facility at the Zhukovsky Air Development Center near Moscow. The centerpiece of the research program was the Tu 144LL, a first-generation Russian supersonic jetliner that was modified by its developer/builder, Tupolev ANTK (aviatsionnyy nauchno-tekhnicheskiy kompleks-roughly, aviation technical complex), into a flying laboratory for supersonic research. Using the Tu-144LL to conduct flight research experiments, researchers compared full-scale supersonic aircraft flight data with results from models in wind tunnels, computer-aided techniques, and other flight tests. The experiments provided unique aerodynamic, structures, acoustics, and operating environment data on supersonic passenger aircraft. Data collected from the research program was being used to develop the technology base for a proposed future American-built supersonic jetliner. Although actual development of such an advanced supersonic transport (SST) is currently on hold, commercial aviation experts estimate that a market for up to 500 such aircraft could develop by the third decade of the 21st Century. The Tu-144LL used in the NASA-sponsored research program was a 'D' model with different engines than were used in production

  15. The gammaTuRC Nanomachine Mechanism and Future Applications

    NASA Astrophysics Data System (ADS)

    Riehlman, Timothy D.

    The complexity and precision of the eukaryotic cell's cytoskeletal network is unrivaled by any man-made systems, perfected by billions of years of evolution, mastering elegant processes of self-assembly, error correction, and self-repair. Understanding the capabilities of these networks will have important and far reaching applications in human medicine by aiding our understanding of developmental processes, cellular division, and disease mechanisms, and through biomimicry will provide insights for biosynthetic manufacturing at the nanoscale and across scales. My research utilizes cross species techniques from Human to the model organism of Fission Yeast to investigate the structure and mechanisms of the g-tubulin ring complex (gTuRC). The gTuRC is a highly conserved eukaryotic multiprotein complex serving as a microtubule organizing center (MTOC) responsible for microtubule nucleation through templating, regulation of dynamics, and establishment of microtubule polarity. Microtubules are 25 nm diameter dynamic flexible polymers of a/b-tubulin heterodimers that function as scaffolds, force generators, distributors, and intracellular highways. The microtubule cytoskeleton is essential for numerous fundamental cellular processes such as mitotic division of chromosomes and cell division, organelle distribution within the cell, cell signaling, and cell shape. This incredible diversity in functions is made possible in part due to molecular motor Kinesin-like proteins (Klps), which allow expansion into more specialized neural, immune, and ciliated cell functions. Combined, the MTOC, microtubules, and Klps represent ideal microtubule cytoskeleton protein (MCP) modular components for in vitro biomimicry towards generation of adaptable patterned networks for human designed applications. My research investigates the hypothesis that a mechanistic understanding of conserved MTOC gTuRC mechanisms will help us understand dynamic cellular nanomachines and their ability to self

  16. Sphingosine 1-phosphate lyase inhibition by 2-acetyl-4-(tetrahydroxybutyl)imidazole (THI) under conditions of vitamin B6 deficiency.

    PubMed

    Ohtoyo, Mamoru; Tamura, Masakazu; Machinaga, Nobuo; Muro, Fumihito; Hashimoto, Ryuji

    2015-02-01

    Caramel food colorant 2-acetyl-4-(tetrahydroxybutyl)imidazole (THI) causes lymphopenia in animals through sphingosine 1-phosphate lyase (SPL) inhibition. However, this mechanism of action is partly still controversial because THI did not inhibit SPL in vitro either in cell-free or in cell-based systems. It is thought that the in vitro experimental conditions which have been used so far were not suitable for the evaluation of SPL inhibition, especially in case of cell-based experiments. We speculated that the key factor might be the coenzyme pyridoxal 5'-phosphate (PLP), an active form of vitamin B6 (VB6), because media used in cell-based assays usually contain an excess amount of VB6 which leads to the activation of SPL. By the use of VB6-deficient culture medium, we could regulate apo- (without PLP) and holo- (with PLP) SPL enzyme in cultured cells, resulting in the successful detection of SPL inhibition by THI. Although the observed inhibitory effect was not as strong as that of 4-deoxypyridoxine (a VB6 analog SPL inhibitor), these findings may be useful for further understanding the mechanism of action of THI. PMID:25381637

  17. Value of tissue harmonic imaging (THI) and contrast harmonic imaging (CHI) in detection and characterisation of breast tumours

    PubMed Central

    Jung, E. M.; Jungius, K.-P.; Ertan, K.

    2006-01-01

    The purpose of this study was to investigate the extent to which tissue harmonic imaging (THI), speckle reduction imaging (SRI), spatial compounding (SC) and contrast can improve detection and differentiation of breast tumours. We examined 38 patients (14 benign, 24 malignant tumours) with different combinations of THI, SRI and SC. The effect on delineation, margin, tissue differentiation and posttumoral phenomena was evaluated with a three-point score. Additionally, 1oo not palpable tumours (diameters: 4–15 mm) were examined by contrast harmonic imaging (CHI) with power Doppler. After bolus injection (0.5 ml Optison), vascularisation and enhancement were observed for 20 min. The best combination for detection of margin, infiltration, echo pattern and posterior lesion boundary was the combination of SRI level 2 with SC low. THI was helpful for lesions OF more than 1 cm depth. In native Power Doppler, vessels were found in 54 of 100 lesions. Within 5 min after contrast medium (CM) injection, marginal and penetrating vessels increased in benign and malignant tumours and central vessels mostly in carcinomas (p<0.05). A diffuse CM accumulation was observed up to 20 min after injection in malignant tumours only (p<0.05). THI, SRI and SC improved delineation and tissue differentiation. Second-generation contrast agent allowed detection of tumour vascularisation with prolonged enhancement. PMID:16823568

  18. Differential induction of the Arabidopsis thaliana Thi2.1 gene by Fusarium oxysporum f. sp. matthiolae.

    PubMed

    Epple, P; Vignutelli, A; Apel, K; Bohlmann, H

    1998-06-01

    The Arabidopsis thaliana Thi2.1 gene is inducible by necrotrophic fungi through a signal transduction pathway different from that for pathogenesis-related (PR) proteins. We have identified three ecotypes that are susceptible (Col-2, Ler, and Ws) and two ecotypes that are resistant (Mt-0 and Uk-4) to spray inoculation with Fusarium oxysporum f. sp. matthiolae. The Thi2.1 transcript level after infection correlates with resistance, being 5 to 10 times higher in the resistant than in the susceptible ecotypes. The beta-glucuronidase (GUS) expression of a Thi2.1-promoter-uidA fusion (with a promoter derived from Col-2) is on the average almost 10 times higher in the Uk-4 background than in the Col-2 background. This confirms the results obtained by Northern (RNA) blots and indicates that Uk-4, and probably other resistant ecotypes too, might have a more sensitive recognition system for F. oxysporum f. sp. matthiolae or might have a signal transduction system that gives a higher amplification of the original recognition signal. Our results suggest a role of the Thi2.1 gene in resistance against F. oxysporum f. sp. matthiolae and perhaps other necrotrophic fungi. PMID:9612950

  19. The Measure your World / Mide tu Mundo Project

    NASA Astrophysics Data System (ADS)

    Hojman, S.; Johnson, R. M.; Meymaris, K. K.; Ward, D. L.; Russell, R.; Genyuk, J.; Lagrave, M.; Henderson, S.; Ostrosky, J.; Martinez, M.

    2007-12-01

    Over 22 centuries ago Erathostenes devised a method to determine the Earth radius. Measure your World / Mide tu Mundo is a joint venture lead by Windows to the Universe/Ventanas al Universo in the United States of America, EducaRed in Chile and RedEscolar in Mexico seeking to partner teams of students, teachers and parents from the three countries in a collaborative effort to determine the Earth circumference by sharing the results of their measurements, and exchanging cultural information. Data are collected from September 29 through October 7, 2007. A report of the whole experience will be presented.

  20. A comparison of THI indices leads to a sensible heat-based heat stress index for shaded cattle that aligns temperature and humidity stress

    NASA Astrophysics Data System (ADS)

    Berman, A.; Horovitz, Talia; Kaim, M.; Gacitua, H.

    2016-01-01

    The combined temperature-humidity heat stress is estimated in farm animals by indices derived of an index based on human thermal comfort sensation. The latter index consists of temperature and humidity measures that sum to form the temperature-humidity index (THI). The hitherto unknown relative contribution of temperature and humidity to the THI was examined. A temperature-humidity data set (temperature 20-42 °C and relative humidity 10-70 %) was used to assess by regression procedures the relative weights of temperature and humidity in the variance of THI values produced by six commonly used heat stress indices. The temperature (Ta) effect was predominant (0.82-0.95 of variance) and humidity accounted for only 0.05 to 0.12 of THI variance, half of the variance encountered in animal responses to variable humidity heat stress. Significant difference in THI values was found between indices in the relative weights of temperature and humidity. As in THI indices, temperature and humidity are expressed in different physical units, their sum has no physical attributes, and empirical evaluations assess THI relation to animal responses. A sensible heat THI was created, in which at higher temperatures humidity reaches 0.25 of sensible heat, similarly to evaporative heat loss span in heat stressed animals. It relates to ambient temperature-humidity similarly to present THI; its values are similar to other THI but greater at higher humidity. In warm conditions, mean animal responses are similar in both indices. The higher sensitivity to humidity makes this index preferable for warm-humid conditions.

  1. Increase in furfural tolerance in ethanologenic Escherichia coli LY180 by plasmid-based expression of thyA.

    PubMed

    Zheng, Huabao; Wang, Xuan; Yomano, Lorraine P; Shanmugam, Keelnatham T; Ingram, Lonnie O

    2012-06-01

    Furfural is an inhibitory side product formed during the depolymerization of hemicellulose by mineral acids. Genomic libraries from three different bacteria (Bacillus subtilis YB886, Escherichia coli NC3, and Zymomonas mobilis CP4) were screened for genes that conferred furfural resistance on plates. Beneficial plasmids containing the thyA gene (coding for thymidylate synthase) were recovered from all three organisms. Expression of this key gene in the de novo pathway for dTMP biosynthesis improved furfural resistance on plates and during fermentation. A similar benefit was observed by supplementation with thymine, thymidine, or the combination of tetrahydrofolate and serine (precursors for 5,10-methylenetetrahydrofolate, the methyl donor for ThyA). Supplementation with deoxyuridine provided a small benefit, and deoxyribose was of no benefit for furfural tolerance. A combination of thymidine and plasmid expression of thyA was no more effective than either alone. Together, these results demonstrate that furfural tolerance is increased by approaches that increase the supply of pyrimidine deoxyribonucleotides. However, ThyA activity was not directly affected by the addition of furfural. Furfural has been previously shown to damage DNA in E. coli and to activate a cellular response to oxidative damage in yeast. The added burden of repairing furfural-damaged DNA in E. coli would be expected to increase the cellular requirement for dTMP. Increased expression of thyA (E. coli, B. subtilis, or Z. mobilis), supplementation of cultures with thymidine, and supplementation with precursors for 5,10-methylenetetrahydrofolate (methyl donor) are each proposed to increase furfural tolerance by increasing the availability of dTMP for DNA repair. PMID:22504824

  2. Heat-induced Accumulation of Chloroplast Protein Synthesis Elongation Factor, EF-TU, in Winter Wheat

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Chloroplast protein synthesis elongation factor, EF-Tu, has been implicated in heat tolerance in maize (Zea mays L.). Chloroplast EF-Tu is highly conserved, and it is possible that this protein may be of importance to heat tolerance in other species including wheat (Triticum aestivum L.). In this ...

  3. Co-ordination of Mobile Information Agents in TuCSoN.

    ERIC Educational Resources Information Center

    Omicini, Andrea; Zambonelli, Franco

    1998-01-01

    Examines mobile agent coordination and presents TuCSoN, a coordination model for Internet applications based on mobile information agents that uses a tuple centre, a tuple space enhanced with the capability of programming its behavior in response to communication events. Discusses the effectiveness of the TuCSoN model in the contexts of Internet…

  4. TLR4-dependent activation of inflammatory cytokine response in macrophages by Francisella elongation factor Tu1

    PubMed Central

    Sharma, Jyotika; Mishra, Bibhuti B.; Li, Qun; Teale, Judy M.

    2011-01-01

    The bacterial determinants of pulmonary Francisella induced inflammatory responses and their interaction with host components are not clearly defined. In this study, proteomic and immunoblot analyses showed presence of a cytoplasmic protein elongation factor Tu (EF-Tu) in the membrane fractions of virulent F. novicida, LVS and SchuS4, but not in an attenuated F. novicida mutant. EF-Tu was immunodominant in mice vaccinated and protected from virulent F. novicida. Moreover, recombinant EF-Tu induced macrophages to produce inflammatory cytokines in a TLR4 dependent manner. This study shows immune stimulatory properties of a cytoplasmic protein EF-Tu expressed on the membrane of virulent Francisella strains. PMID:21497800

  5. THY-1 Receptor Expression Differentiates Cardiosphere-Derived Cells with Divergent Cardiogenic Differentiation Potential

    PubMed Central

    Gago-Lopez, Nuria; Awaji, Obinna; Zhang, Yiqiang; Ko, Christopher; Nsair, Ali; Liem, David; Stempien-Otero, April; MacLellan, W. Robb

    2014-01-01

    Summary Despite over a decade of intense research, the identity and differentiation potential of human adult cardiac progenitor cells (aCPC) remains controversial. Cardiospheres have been proposed as a means to expand aCPCs in vitro, but the identity of the progenitor cell within these 3D structures is unknown. We show that clones derived from cardiospheres could be subdivided based on expression of thymocyte differentiation antigen 1 (THY-1/CD90) into two distinct populations that exhibit divergent cardiac differentiation potential. One population, which is CD90+, expressed markers consistent with a mesenchymal/myofibroblast cell. The second clone type was CD90− and could form mature, functional myocytes with sarcomeres albeit at a very low rate. These two populations of cardiogenic clones displayed distinct cell surface markers and unique transcriptomes. Our study suggests that a rare aCPC exists in cardiospheres along with a mesenchymal/myofibroblast cell, which demonstrates incomplete cardiac myocyte differentiation. PMID:24936447

  6. The elongation factor Tu.kirromycin complex has two binding sites for tRNA molecules.

    PubMed Central

    van Noort, J M; Duisterwinkel, F J; Jonák, J; Sedlácek, J; Kraal, B; Bosch, L

    1982-01-01

    The interaction of the polypeptide chain elongation factor Tu (EF-Tu) with the antibiotic kirromycin and tRNA has been studied by measuring the extent of protein modification with N-tosyl-L-phenylalanine chloromethylketone (TPCK) and N-ethylmaleimide (NEM). Kirromycin protects both EF-Tu.GDP and EF-Tu.GTP against modification with TPCK. Binding of aminoacyl-tRNA added at increasing concentrations to a solution of 40 microM EF-Tu.GDP.kirromycin complex re-exposes the TPCK target site on the protein. However, when the aminoacyl-tRNA concentration is raised beyond 20 microM, TPCK labeling drops again and is blocked completely at approximately 300 microM aminoacyl-tRNA. By contrast, addition of uncharged tRNA or N- acetylaminoacyl -tRNA enhances TPCK labeling of the protein over the entire tRNA concentration range studied. These data strongly suggest that kirromycin induces in EF-Tu.GDP an additional tRNA binding site that can bind uncharged tRNA, aminoacyl-tRNA, and N- acetylaminoacyl -tRNA. Support for this assumption is provided by measuring the modification of EF-Tu.GDP with the sulfhydryl reagent NEM. Moreover, NEM modification also indicates an additional tRNA binding site on EF-Tu.GTP.kirromycin, which could not be detected with TPCK. Mapping of the tryptic peptides of EF-Tu.GDP labeled with [14C]TPCK revealed only one target site for this agent, i.e., cysteine-81. Modification occurred at the same site in the presence and in the absence of kirromycin and uncharged tRNA.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:6765192

  7. Immune Reconstitution after Autologous Hematopoietic Transplantation with Lin−, CD34+, Thy-1LO Selected or Intact Stem Cell Products*

    PubMed Central

    Singh, Rakesh K.; Varney, Michelle L.; Leutzinger, Cheryl; Vose, Julie M.; Bierman, Philip J.; Buyukberber, Suleyman; Ino, Kazuhiko; Loh, Kevin; Nichols, Craig; Inwards, David; Rifkin, Robert; Talmadge, James E.

    2007-01-01

    In sequential studies, we compared immune reconstitution following high dose chemotherapy (HDT) and stem cell transplantation (SCT) using intact mobilized peripheral blood stem cell (PSC) in intermediate grade non-Hodgkin's lymphoma (NHL) patients and CD34+, lineage negative (Lin−), Thy-1lo (CD34+Lin−Thy-1lo) stem cells in low-grade NHL patients. Cytokine expression and cellular phenotype and function were used as the basis for comparison. Despite differences in cellular composition of the stem cell grafts, immune reconstitution in both groups was similar. Significantly higher levels of type 1 and 2-associated cytokine messenger ribonucleic acid (mRNA) were observed both prior to and following transplant in the peripheral blood (PB) of both cohorts as compared to normal individuals. Similar levels of interleukin (IL)-4, IL-10, interferon-gamma (IFN-γ), and tumor necrosis factor-alpha (TNF-α) mRNA were seen in PB mononuclear cells following transplant with either product. In contrast, patients receiving isolated CD34+Lin−Thy-1lo cells expressed significantly higher IL-2 levels at all times examined post-transplant. Despite the high levels of cytokine gene expression and rapid restoration to pretransplant levels of CD3 cell number by day 30, T cell function and CD4:CD8 and CD4+CD45RA:CD4+CD45RO+ ratios were significantly depressed in both cohorts compared to normal donors, and significantly lower in patients transplanted with CD34+Lin−Thy-1lo compared to patients receiving an intact PSC product. These data suggest that the peripheral tolerance in patients receiving HDT and an autologous SCT occurs independent of graft composition, although immune function and CD4 recovery is better facilitated by transplantation of an intact product. PMID:17570320

  8. Cyan fluorescent protein expression in ganglion and amacrine cells in a thy1-CFP transgenic mouse retina

    PubMed Central

    Vila, Alejandro; Huynh, Uyen-Chi N.; Brecha, Nicholas C.

    2008-01-01

    Purpose To characterize cyan fluorescent protein (CFP) expression in the retina of the thy1-CFP (B6.Cg-Tg(Thy1-CFP)23Jrs/J) transgenic mouse line. Methods CFP expression was characterized using morphometric methods and immunohistochemistry with antibodies to neurofilament light (NF-L), neuronal nuclei (NeuN), POU-domain protein (Brn3a) and calretinin, which immunolabel ganglion cells, and syntaxin 1 (HPC-1), glutamate decarboxylase 67 (GAD67), GABA plasma membrane transporter-1 (GAT-1), and choline acetyltransferase (ChAT), which immunolabel amacrine cells. Results CFP was extensively expressed in the inner retina, primarily in the inner plexiform layer (IPL), ganglion cell layer (GCL), nerve fiber layer, and optic nerve. CFP fluorescent cell bodies were in all retinal regions and their processes ramified in all laminae of the IPL. Some small, weakly CFP fluorescent somata were in the inner nuclear layer (INL). CFP-containing somata in the GCL ranged from 6 to 20 μm in diameter, and they had a density of 2636±347 cells/mm2 at 1.5 mm from the optic nerve head. Immunohistochemical studies demonstrated colocalization of CFP with the ganglion cell markers NF-L, NeuN, Brn3a, and calretinin. Immunohistochemistry with antibodies to HPC-1, GAD67, GAT-1, and ChAT indicated that the small, weakly fluorescent CFP cells in the INL and GCL were cholinergic amacrine cells. Conclusions The total number and density of CFP-fluorescent cells in the GCL were within the range of previous estimates of the total number of ganglion cells in the C57BL/6J line. Together these findings suggest that most ganglion cells in the thy1-CFP mouse line 23 express CFP. In conclusion, the thy1-CFP mouse line is highly useful for studies requiring the identification of ganglion cells. PMID:18728756

  9. Loss of medial septum cholinergic neurons in THY-Tau22 mouse model: what links with tau pathology?

    PubMed

    Belarbi, K; Burnouf, S; Fernandez-Gomez, F-J; Desmercières, J; Troquier, L; Brouillette, J; Tsambou, L; Grosjean, M-E; Caillierez, R; Demeyer, D; Hamdane, M; Schindowski, K; Blum, D; Buée, L

    2011-09-01

    Alzheimer's disease (AD) is a neurodegenerative disorder histologically defined by the cerebral accumulation of amyloid deposits and neurofibrillary tangles composed of hyperphosphorylated tau proteins. Loss of basal forebrain cholinergic neurons is another hallmark of the disease thought to contribute to the cognitive dysfunctions. To this date, the mechanisms underlying cholinergic neurons degeneration remain uncertain. The present study aimed to investigate the relationship between neurofibrillary degeneration and cholinergic defects in AD using THY-Tau22 transgenic mouse model exhibiting a major hippocampal AD-like tau pathology and hyperphosphorylated tau species in the septohippocampal pathway. Here, we report that at a time THY-Tau22 mice display strong reference memory alterations, the retrograde transport of fluorogold through the septohippocampal pathway is altered. This impairment is associated with a significant reduction in the number of choline acetyltransferase (ChAT)-immunopositive cholinergic neurons in the medial septum. Analysis of nerve growth factor (NGF) levels supports an accumulation of the mature neurotrophin in the hippocampus of THY-Tau22 mice, consistent with a decrease of its uptake or retrograde transport by cholinergic terminals. Finally, our data strongly support that tau pathology could be instrumental in the cholinergic neuronal loss observed in AD. PMID:21605043

  10. Host sex-dependent growth of potential Thy-1+ lymphoma cells that appear in the thymus of X-irradiated NFS mice

    SciTech Connect

    Mori, N.; Takamori, Y. )

    1990-01-01

    During the course of studies designed to detect potentially leukemic cells in radiation lymphomagenesis, using an opposite-sex (male----female) transplantation assay method, we previously found that potential Thy-1- lymphoma cells are generated in the bone marrow of NFS mice exposed to a split-dose irradiation (1.7 Gy X 4), while potential Thy-1+ lymphoma cells are not detectable. In this report, using a (female----male) intrathymic transplantation assay system we show that potential Thy-1+ lymphoma cells were generated in the thymus of female NFS mice exposed to split-dose irradiation, and reconfirm that such cells were not detected in the (male----female) transplantation system. These results demonstrate that the detection of potential Thy-1+ lymphoma cells strictly depends on the transplantation system.

  11. An unusual mechanism of thymidylate biosynthesis in organisms containing the thyX Gene

    PubMed Central

    Koehn, Eric M.; Fleischmann, Todd; Conrad, John A.; Palfey, Bruce A.; Lesley, Scott A.; Mathews, Irimpan I.; Kohen, Amnon

    2009-01-01

    Biosynthesis of the DNA base thymine depends on activity of the enzyme thymidylate synthase (TS) to catalyze the methylation of the uracil moiety of 2’-deoxyuridine-5’-monophosphate (dUMP). All known thymidylate synthases (TSs) rely on an active site residue of the enzyme to activate dUMP1, 2. This functionality has been demonstrated for classical TSs, including human TS, and is instrumental in mechanism-based inhibition of these enzymes. Here we report the first example of thymidylate biosynthesis that occurs without an enzymatic nucleophile. This unusual biosynthetic pathway occurs in organisms containing the thyX gene, which codes for a flavin-dependent thymidylate synthase (FDTS), and is present in several human pathogens3–5. Our findings indicate that the putative active site nucleophile is not required for FDTS catalysis, and no alternative nucleophilic residues capable of serving this function can be identified. Instead, our findings suggest that a hydride equivalent (i.e. a proton and two electrons) is transferred from the reduced flavin cofactor directly to the uracil ring, followed by an isomerization of the intermediate to form the product, 2’-deoxythymidine-5’-monophosphate (dTMP). These observations indicate a very different chemical cascade than that of classical TSs or any other known biological methylation. The findings and chemical mechanism proposed here, together with available structural data, suggest that selective inhibition of FDTSs, with little effect on human thymine biosynthesis, should be feasible. Since several human pathogens depend on FDTS for DNA biosynthesis, its unique mechanism makes it an attractive target for antibiotic drugs. PMID:19370033

  12. Hypoxia-induced DNA hypermethylation in human pulmonary fibroblasts is associated with Thy-1 promoter methylation and the development of a pro-fibrotic phenotype

    PubMed Central

    2012-01-01

    Background Pulmonary fibrosis is a debilitating and lethal disease with no effective treatment options. Understanding the pathological processes at play will direct the application of novel therapeutic avenues. Hypoxia has been implicated in the pathogenesis of pulmonary fibrosis yet the precise mechanism by which it contributes to disease progression remains to be fully elucidated. It has been shown that chronic hypoxia can alter DNA methylation patterns in tumour-derived cell lines. This epigenetic alteration can induce changes in cellular phenotype with promoter methylation being associated with gene silencing. Of particular relevance to idiopathic pulmonary fibrosis (IPF) is the observation that Thy-1 promoter methylation is associated with a myofibroblast phenotype where loss of Thy-1 occurs alongside increased alpha smooth muscle actin (α-SMA) expression. The initial aim of this study was to determine whether hypoxia regulates DNA methylation in normal human lung fibroblasts (CCD19Lu). As it has been reported that hypoxia suppresses Thy-1 expression during lung development we also studied the effect of hypoxia on Thy-1 promoter methylation and gene expression. Methods CCD19Lu were grown for up to 8 days in hypoxia and assessed for global changes in DNA methylation using flow cytometry. Real-time PCR was used to quantify expression of Thy-1, α-SMA, collagen I and III. Genomic DNA was bisulphite treated and methylation specific PCR (MSPCR) was used to examine the methylation status of the Thy-1 promoter. Results Significant global hypermethylation was detected in hypoxic fibroblasts relative to normoxic controls and was accompanied by increased expression of myofibroblast markers. Thy-1 mRNA expression was suppressed in hypoxic cells, which was restored with the demethylating agent 5-aza-2′-deoxycytidine. MSPCR revealed that Thy-1 became methylated following fibroblast exposure to 1% O2. Conclusion These data suggest that global and gene-specific changes in

  13. Inhibition of human pyridoxal kinase by 2-acetyl-4-((1R,2S,3R)-1,2,3,4-tetrahydroxybutyl)imidazole (THI).

    PubMed

    Elsinghorst, Paul W; di Salvo, Martino L; Parroni, Alessia; Contestabile, Roberto

    2015-04-01

    2-Acetyl-4-((1R,2S,3R)-1,2,3,4-tetrahydroxybutyl)imidazole (THI) is observed as a minor contaminant in caramel food colourings (E 150c). Feeding experiments with rodents have revealed a significant lymphopenic effect that has been linked to the presence of THI in these food colourings. Pyridoxal kinase inhibition by THI has been suggested, but not demonstrated, as a mode of action as it leads to lowered levels of pyridoxal-5'-phosphate, which are known to cause lymphopenia. Recently, THI was also shown to inhibit sphingosine-1-phosphate lyase causing comparable immunosuppressive effects and derivatives of THI are being developed for the treatment of rheumatoid arthritis in humans. Interestingly, sphingosine-1-phosphate lyase activity depends on pyridoxal-5'-phosphate, which in turn is provided by pyridoxal kinase. This report shows that THI does inhibit pyridoxal kinase with competitive and mixed-type non-competitive behaviour towards its two substrates, pyridoxal and ATP, respectively. The corresponding inhibition constants are in the low millimolar range. PMID:24899377

  14. Extensive genetic heterogeneity in the neuroblastoma cell line NB(TU)1.

    PubMed

    Inoue, A; Yokomori, K; Tanabe, H; Mizusawa, H; Sofuni, T; Hayashi, Y; Tsuchida, Y; Shimatake, H

    1997-09-17

    A neuroblastoma cell line displaying genetically unique features was established from a stage III case of a 20-month-old girl. Southern blotting by the probe pTNB6, which contains exon 1 of the N-myc gene, showed that the primary tumor had in total 4 aberrant bands beside the normal amplified band. The established cell line NB(TU)1 had an aberrant N-myc band (9.0 kb) in addition to the normal band (2.9 kb). Cytogenetic analysis revealed that NB(TU)1 has a composite karyotype composed of at least 7 related karyotypes, which are pseudo-diploid and contain complex chromosomal abnormalities, including translocations, deletions and homogeneously staining regions (HSRs). Such extensive abnormalities were considered to be prominent among known neuroblastoma cell lines, and it was suggested that NB(TU)1 had acquired a certain type of genetic instability. Analysis of N-myc bands in 11 clones of NB(TU)1 showed that the intensity ratio of the normal-sized band (2.9 kb) and the aberrant one (9.0 kb) markedly varied among clones. Moreover, 3 clones showed an additional band with the size of 3.7 kb, which was detectable neither in the parent NB(TU)1 nor in the primary tumor. Thus, NB(TU)1 was shown to be composed of heterogeneous cell components. To further detect such ongoing chromosomal instability, we examined micronuclei formation. NB(TU)1 yielded a larger number of micronuclei than 5 other neuroblastoma cell lines. We conclude that NB(TU)1 has acquired genetic instability detectable by both Southern blotting and cytogenetic analysis. PMID:9378542

  15. Genetic control of immunity to Turnip mosaic virus (TuMV) pathotype 1 in Brassica rapa (Chinese cabbage).

    PubMed

    Lydiate, Derek J; Pilcher, Rachel L Rusholme; Higgins, Erin E; Walsh, John A

    2014-08-01

    Turnip mosaic virus (TuMV) is the major virus infecting crops of the genus Brassica worldwide. A dominant resistance gene, TuRB01b, that confers immunity to the virus isolate UK 1 (a representative pathotype 1 isolate of TuMV) on Brassica rapa was identified in the Chinese cabbage cultivar Tropical Delight. The TuRB01b locus was mapped to a 2.9-cM interval on B. rapa chromosome 6 (A6) that was flanked by RFLP markers pN101e1 and pW137e1. This mapping used a first backcross (B(1)) population segregating for the resistance gene at TuRB01b and sets of RFLP markers employed in previous mapping experiments in Brassica. Virus-plant interaction phenotypes were assayed in inbred progeny derived from B(1) individuals to allow different virus isolates to be tested. Comparative mapping confirmed that A6 of B. rapa was equivalent to chromosome 6 of Brassica napus (A6) and that the map position of TuRB01b in B. rapa could be identical to that of TuRB01 in B. napus. Detailed evaluation of plant-virus interactions showed that TuRB01 and TuRB01b had indistinguishable specificities to a range of TuMV isolates. The possibility that TuRB01 and TuRB01b represent similar or identical alleles at the same A genome resistance locus suggests that B. napus acquired TuRB01 from the B. rapa gene pool. PMID:25275757

  16. Cell-cycle dependent phosphorylation of yeast pericentrin regulates γ-TuSC-mediated microtubule nucleation.

    PubMed

    Lin, Tien-Chen; Neuner, Annett; Schlosser, Yvonne T; Scharf, Annette N D; Weber, Lisa; Schiebel, Elmar

    2014-01-01

    Budding yeast Spc110, a member of γ-tubulin complex receptor family (γ-TuCR), recruits γ-tubulin complexes to microtubule (MT) organizing centers (MTOCs). Biochemical studies suggest that Spc110 facilitates higher-order γ-tubulin complex assembly (Kollman et al., 2010). Nevertheless the molecular basis for this activity and the regulation are unclear. Here we show that Spc110 phosphorylated by Mps1 and Cdk1 activates γ-TuSC oligomerization and MT nucleation in a cell cycle dependent manner. Interaction between the N-terminus of the γ-TuSC subunit Spc98 and Spc110 is important for this activity. Besides the conserved CM1 motif in γ-TuCRs (Sawin et al., 2004), a second motif that we named Spc110/Pcp1 motif (SPM) is also important for MT nucleation. The activating Mps1 and Cdk1 sites lie between SPM and CM1 motifs. Most organisms have both SPM-CM1 (Spc110/Pcp1/PCNT) and CM1-only (Spc72/Mto1/Cnn/CDK5RAP2/myomegalin) types of γ-TuCRs. The two types of γ-TuCRs contain distinct but conserved C-terminal MTOC targeting domains.DOI: http://dx.doi.org/10.7554/eLife.02208.001. PMID:24842996

  17. Joint US/Russia TU-144 Engine Ground Tests

    NASA Technical Reports Server (NTRS)

    Acosta, Waldo A.; Balser, Jeffrey S.; McCartney, Timothy P.; Richter, Charles A.; Woike, Mark R.

    1997-01-01

    Two engine research experiments were recently completed in Moscow, Russia using an engine from the Tu-144 supersonic transport airplane. This was a joint project between the United States and Russia. Personnel from the NASA Lewis Research Center, General Electric Aircraft Engines, Pratt & Whitney, the Tupolev Design Bureau, and EBP Aircraft LTD worked together as a team to overcome the many technical and cultural challenges. The objective was to obtain large scale inlet data that could be used in the development of a supersonic inlet system for a future High Speed Civil Transport (HSCT). The-first experiment studied the impact of typical inlet structures that have trailing edges in close proximity to the inlet/engine interface plane on the flow characteristics at that plane. The inlet structure simulated the subsonic diffuser of a supersonic inlet using a bifurcated splitter design. The centerbody maximum diameter was designed to permit choking and slightly supercritical operation. The second experiment measured the reflective characteristics of the engine face to incoming perturbations of pressure amplitude. The basic test rig from the first experiment was used with a longer spacer equipped with fast actuated doors. All the objectives set forth at the beginning of the project were met.

  18. A Thy1-CFP DBA/2J mouse line with cyan fluorescent protein expression in retinal ganglion cells

    PubMed Central

    RAYMOND, IONA D.; POOL, ANGELA L.; VILA, ALEJANDRO; BRECHA, NICHOLAS C.

    2013-01-01

    A DBA/2J (D2) transgenic mouse line with cyan fluorescent protein (CFP) reporter expression in ganglion cells was developed for the analysis of ganglion cells during progressive glaucoma. The Thy1-CFP D2 (CFP-D2) line was created by congenically breeding the D2 line, which develops pigmentary glaucoma, and the Thy1-CFP line, which expresses CFP in ganglion cells. Microsatellite marker analysis of CFP-D2 progeny verified the genetic inclusion of the D2 isa and ipd loci. Specific mutations within these loci lead to dysfunctional melanosomal proteins and glaucomatous phenotype in D2 mice. Polymerase chain reaction analysis confirmed the inclusion of the Thy1-CFP transgene. CFP-fluorescent ganglion cells, 6–20 μm in diameter, were distributed in all retinal regions, CFP processes were throughout the inner plexiform layer, and CFP-fluorescent axons were in the fiber layer and optic nerve head. Immunohistochemistry with antibodies to ganglion cell markers NF-L, NeuN, Brn3a, and SMI32 was used to confirm CFP expression in ganglion cells. Immunohistochemistry with antibodies to amacrine cell markers HPC-1 and ChAT was used to confirm weak CFP expression in cholinergic amacrine cells. CFP-D2 mice developed a glaucomatous phenotype, including iris disease, ganglion cell loss, attrition of the fiber layer, and elevated intraocular pressure. A CFP-D2 transgenic line with CFP-expressing ganglion cells was developed, which has (1) a predominantly D2 genetic background, (2) CFP-expressing ganglion cells, and (3) age-related progressive glaucoma. This line will be of value for experimental studies investigating ganglion cells and their axons in vivo and in vitro during the progressive development of glaucoma. PMID:19930759

  19. ATP release due to Thy-1–integrin binding induces P2X7-mediated calcium entry required for focal adhesion formation

    PubMed Central

    Henríquez, Mauricio; Herrera-Molina, Rodrigo; Valdivia, Alejandra; Alvarez, Alvaro; Kong, Milene; Muñoz, Nicolás; Eisner, Verónica; Jaimovich, Enrique; Schneider, Pascal; Quest, Andrew F. G.; Leyton, Lisette

    2011-01-01

    Thy-1, an abundant mammalian glycoprotein, interacts with αvβ3 integrin and syndecan-4 in astrocytes and thus triggers signaling events that involve RhoA and its effector p160ROCK, thereby increasing astrocyte adhesion to the extracellular matrix. The signaling cascade includes calcium-dependent activation of protein kinase Cα upstream of Rho; however, what causes the intracellular calcium transients required to promote adhesion remains unclear. Purinergic P2X7 receptors are important for astrocyte function and form large non-selective cation pores upon binding to their ligand, ATP. Thus, we evaluated whether the intracellular calcium required for Thy-1-induced cell adhesion stems from influx mediated by ATP-activated P2X7 receptors. Results show that adhesion induced by the fusion protein Thy-1-Fc was preceded by both ATP release and sustained intracellular calcium elevation. Elimination of extracellular ATP with Apyrase, chelation of extracellular calcium with EGTA, or inhibition of P2X7 with oxidized ATP, all individually blocked intracellular calcium increase and Thy-1-stimulated adhesion. Moreover, Thy-1 mutated in the integrin-binding site did not trigger ATP release, and silencing of P2X7 with specific siRNA blocked Thy-1-induced adhesion. This study is the first to demonstrate a functional link between αvβ3 integrin and P2X7 receptors, and to reveal an important, hitherto unanticipated, role for P2X7 in calcium-dependent signaling required for Thy-1-stimulated astrocyte adhesion. PMID:21502139

  20. Thy-1-mediated Cell -Cell Contact Induces Astrocyte Migration through the Engagement of αVβ3 Integrin and Syndecan-4

    PubMed Central

    Kong, Milene; Muñoz, Nicolás; Valdivia, Alejandra; Alvarez, Alvaro; Herrera-Molina, Rodrigo; Cárdenas, Areli; Schneider, Pascal; Burridge, Keith; Quest, Andrew F. G.; Leyton, Lisette

    2013-01-01

    Cell adhesion to the extracellular matrix (ECM) proteins occurs through interactions with integrins that bind to Arg-Gly-Asp (RGD) tripeptides, and syndecan-4, which recognizes the heparin-binding domain (HBD) of other proteins. Both receptors trigger signaling pathways, including those that activate RhoGTPases such as RhoA and Rac1. This sequence of events modulates cell adhesion to the ECM and cell migration. Using a neuron-astrocyte model, we have reported that the neuronal protein Thy-1 engages αVβ3 integrin and syndecan-4 to induce RhoA activation and strong astrocyte adhesion to their underlying substrate. Thus, because cell-cell interactions and strong cell attachment to the matrix are considered antagonistic to cell migration, we hypothesized that Thy-1 stimulation of astrocytes should preclude cell migration. Here, we studied the effect of Thy-1 expressing neurons on astrocyte polarization and migration using a wound-healing assay and immunofluorescence analysis. Signaling molecules involved were studied by affinity precipitations, western blots and the usage of specific antibodies. Intriguingly, Thy-1 interaction with its two receptors was found to increase astrocyte polarization and migration. The latter events required interactions of these receptors with both the RGD-like sequence and the HBD of Thy-1. Additionally, prolonged Thy-1-receptor interactions inhibited RhoA activation while activating FAK, PI3K and Rac1. Therefore, sustained engagement of integrin and syndecan-4 with the neuronal surface protein Thy-1 induces astrocyte migration. Interestingly we identify here, a cell-cell interaction that although initially induces strong cell attachment, upon persistant stimulation favors cell migration by engaging the same signaling receptors and molecules as those utilized by ECM proteins to stimulate cell movement. PMID:23481656

  1. The Thermal Hyperspectral Imager (THI): an instrument for remote sensing of Earth's surface from a micro-satellite platform

    NASA Astrophysics Data System (ADS)

    Wright, R.; Lucey, P. G.; Horton, K. A.; Wood, M.; Garbeil, H.; Crites, S. T.

    2011-12-01

    The Thermal Hyperspectral Imager (THI) is a low cost, low mass, power efficient instrument designed to acquire hyperspectral remote sensing data in the long-wave infrared. The instrument has been designed to satisfy mass, volume, and power constraints necessary to allow for its accommodation in a 95 kg micro-satellite bus, designed by staff and students at the University of Hawai'i. THI acquires approximately 30 separate spectral bands in the 8 to 14 μm wavelength region, at 10 wavenumber resolution. Rather than using filtering or dispersion to generate the spectral information, THI uses an interferometric technique. Light from the scene is focused onto an uncooled microbolometer detector array through a stationary interferometer, causing the light incident at each detector at any instant in time to be phase shifted by an optical path difference which varies linearly across the array in the along-track dimension. As platform motion translates the detector array in the along-track direction at a rate of approximately one pixel per frame (the camera acquires data at 30 Hz) the radiance from each scene element can be sampled at each OPD, thus generating an interferogram. Spectral radiance as a function of wavelength is subsequently obtained for each scene element using standard Fourier transform techniques. Housed in a pressure vessel to shield COTS parts from the space environment, the total instrument has a mass of 15 kg. Peak power consumption, largely associated with the calibration procedure, is <90 W. From a nominal altitude of 550 km the resulting data would have a spatial resolution of approximately 300 m. Although an individual imaging event yields approximately 1 Gbit of raw uncompressed data, onboard processing (to convert the interferograms into a conventional spectral hypercube) can reduce this to tens of Mbits per scene. In this presentation we will describe a) the rationale for the project, b) the instrument design, and c) how the data are processed

  2. Calibration Methods for Air Coupled Antennas - COST Action TU1208

    NASA Astrophysics Data System (ADS)

    Marecos, Vânia; Solla, Mercedes; Fontul, Simona; Pajewski, Lara

    2016-04-01

    This work focuses on the comparison of different methods for calibrating air coupled antennas: Coring, Surface Reflection Method (SRM) and Common Mid-Point (CMP) through the analysis of GPR data collected in a test site with different pavement solutions. Research activities have been carried out during a Short Term Scientific Mission (STSM) funded by the COST (European Cooperation in Science and Technology) Action TU1208 "Civil Engineering Applications of Ground Penetrating Radar" in December 2015. The use of GPR in transport infrastructures represents one of the most significant advances for obtaining continuous data along the road, with the advantage of operation at traffic speed and being a non-destructive technique. Its main application has been the evaluation of layer thickness. For the determination of layer thickness, it is necessary to know the velocity of the signal, which depends on the dielectric constant of the material, and the two-way travel time of the reflected signal that is recorded by the GPR system. The calculation of the dielectric value of the materials can be done using different approaches such as: using fixed values based on experience, laboratory determination of dielectric values, applying the SRM, performing back calculation from ground truth references such as cores and test pits, or using the CMP method. The problem with using ground truth is that it is time consuming, labour intensive and intrusive to traffic, in addition, a drill core is not necessarily representative of the whole surveyed area. Regarding the surface reflection technique, one of the problems is that it only measures the dielectric value from the layer surface and not from the whole layer. Recent works already started to address some of these challenges proposing new approaches for GPR layer thickness measurements using multiple antennas to calculate the average dielectric value of the asphalt layer, taking advantage of significant hardware improvements in GPR

  3. Brugia pahangi in nude mice: protective immunity to infective larvae is Thy 1.2+ cell dependent and cyclosporin A resistant.

    PubMed

    Vickery, A C; Nayar, J K

    1987-03-01

    Mechanisms of protective immunity to larvae of Brugia pahangi were studied in congenitally athymic nude C3H/HeN mice and their syngeneic heterozygous littermates. An average 11% of subcutaneous larval inocula was recovered from control nudes 28 days after inoculation. No worms were recovered from nude recipients of viable splenic Thy 1.2+ T lymphocytes from heterozygotes which had killed a priming dose of B. pahangi larvae. Primed T lymphocytes, depleted of either Lyt 1.1+ or Lyt 2.1+ cells or incubated with anti-Thy 1.2 monoclonal antibody and complement, failed to protect nude mice against a larval challenge. Nor were primed B lymphocytes depleted by Thy 1.2+ T cell contaminants protective. Treatment with cyclosporin A (CsA) did not increase the numbers of worms recovered from heterozygotes nor did CsA treatment of heterozygous cell donors abolish the ability of primed Thy 1.2+ T lymphocytes to transfer protection to nude mice. IgG but not IgM antibody titres to B. pahangi antigens were depressed in all CsA-treated mice. CsA treatment of nude mice had no direct effect upon development of B. pahangi larvae. These results show that protective immunity to larvae of B. pahangi in mice depends upon small numbers of Thy 1.2+ T cells which are CsA-resistant. PMID:3494759

  4. Preclinical trials in autosomal dominant AD: Implementation of the DIAN-TU trial

    PubMed Central

    Mills, S.M.; Mallmann, J.; Santacruz, A.M.; Fuqua, A.; Carril, M.; Aisen, P.S.; Althage, M.C.; Belyew, S.; Benzinger, T.L.; Brooks, W.S.; Buckles, V.D.; Cairns, N.J.; Clifford, D.; Danek, A.; Fagan, A.M.; Farlow, M.; Fox, N.; Ghetti, B.; Goate, A.M.; Heinrichs, D.; Hornbeck, R.; Jack, C.; Jucker, M.; Klunk, W.E.; Marcus, D.S.; Martins, R.N.; Masters, C.M.; Mayeux, R.; McDade, E.; Morris, J.C.; Oliver, A.; Ringman, J.M.; Rossor, M.N.; Salloway, S.; Schofield, P.R.; Snider, J.; Snyder, P.; Sperling, R.A.; Stewart, C.; Thomas, R.G.; Xiong, C.; Bateman, R.J.

    2013-01-01

    The Dominantly Inherited Alzheimer’s Network Trials Unit (DIAN-TU) was formed to direct the design and management of interventional therapeutic trials of international DIAN and autosomal dominant Alzheimer’s disease (ADAD) participants. The goal of the DIAN-TU is to implement safe trials that have the highest likelihood of success while advancing scientific understanding of these diseases and clinical effects of proposed therapies. The DIAN-TU has launched a trial design that leverages the existing infrastructure of the ongoing DIAN observational study, takes advantage of a variety of drug targets, incorporates the latest results of biomarker and cognitive data collected during the observational study, and implements biomarkers measuring Alzheimer’s disease (AD) biological processes to improve the efficiency of trial design. The DIAN-TU trial design is unique due to the sophisticated design of multiple drugs, multiple pharmaceutical partners, academics servings as sponsor, geographic distribution of a rare population and intensive safety and biomarker assessments. The implementation of the operational aspects such as home health research delivery, safety magnetic resonance imagings (MRIs) at remote locations, monitoring clinical and cognitive measures, and regulatory management involving multiple pharmaceutical sponsors of the complex DIAN-TU trial are described. PMID:24016464

  5. The surface-associated elongation factor Tu is concealed for antibody binding on viable pneumococci and meningococci.

    PubMed

    Kolberg, Jan; Hammerschmidt, Sven; Frank, Ronald; Jonák, Jirí; Sanderová, Hana; Aase, Audun

    2008-07-01

    Proteome analyses revealed that elongation factor-Tu (EF-Tu) is associated with cytoplasmic membranes of Gram-positive bacteria and outer membranes of Gram-negative bacteria. It is still debatable whether EF-Tu is located on the external side or the internal side of the membranes. Here, we have generated two new monoclonal antibodies (mAbs) and polyclonal rabbit antibodies against pneumococcal EF-Tu. These antibodies were used to investigate the amount of surface-exposed EF-Tu on viable bacteria using a flow cytometric analysis. The control antibodies recognizing the pneumococcal surface protein A and phosphorylcholine showed a significant binding to viable pneumococci. In contrast, anti-EF-Tu antibodies did not recognize pneumococcal EF-Tu. However, heat killing of pneumococci lacking capsular polysaccharides resulted in specific antibody binding to EF-Tu and, moreover, increased the exposure of recognized phosphorylcholine epitopes. Similarly, our EF-Tu-specific antibodies did not recognize EF-Tu of viable Neisseria meningitidis. However, pretreatment of meningococci with ethanol resulted in specific antibody binding to EF-Tu on outer membranes. Importantly, these treatments did not destroy the membrane integrity as analysed with control mAbs directed against cytoplasmic proteins. In conclusion, our flow cytrometric assays emphasize the importance of using viable bacteria and not heat-killed or ethanol-treated bacteria for surface-localization experiments of proteins, because these treatments modulate the cytoplasmic and outer membranes of bacteria and the binding results may not reflect the situation under physiological conditions. PMID:18462389

  6. Selective cytotoxic activity of immunotoxins composed of a monoclonal anti-Thy 1.1 antibody and the ribosome-inactivating proteins bryodin and momordin.

    PubMed Central

    Stirpe, F.; Wawrzynczak, E. J.; Brown, A. N.; Knyba, R. E.; Watson, G. J.; Barbieri, L.; Thorpe, P. E.

    1988-01-01

    The ribosome-inactivating proteins, bryodin, from Bryonia dioica, and momordin, from Momordica charantia, were coupled by a disulphide bond to a monoclonal anti-Thy 1.1 antibody (OX7). Both immunotoxins were specifically cytotoxic to the Thy 1.1-expressing mouse lymphoma cell line AKR-A in vitro. The OX7-bryodin immunotoxins were the more powerfully toxic and reduced protein synthesis in AKR-A cells by 50% at a concentration of 1-4 x 10(-11) M as compared with 1 x 10(-9) M for the OX7-momordin immunotoxins. Neither of the immunotoxins was toxic to mouse lymphoma EL4 cells, which lack the Thy 1.1 antigen, at concentrations up to 3 x 10(-8) M. Further, bryodin and momordin immunotoxins made from an antibody (R10) of irrelevant specificity were without effect on AKR-A cells. PMID:3265330

  7. Selective cytotoxic activity of immunotoxins composed of a monoclonal anti-Thy 1.1 antibody and the ribosome-inactivating proteins bryodin and momordin.

    PubMed

    Stirpe, F; Wawrzynczak, E J; Brown, A N; Knyba, R E; Watson, G J; Barbieri, L; Thorpe, P E

    1988-11-01

    The ribosome-inactivating proteins, bryodin, from Bryonia dioica, and momordin, from Momordica charantia, were coupled by a disulphide bond to a monoclonal anti-Thy 1.1 antibody (OX7). Both immunotoxins were specifically cytotoxic to the Thy 1.1-expressing mouse lymphoma cell line AKR-A in vitro. The OX7-bryodin immunotoxins were the more powerfully toxic and reduced protein synthesis in AKR-A cells by 50% at a concentration of 1-4 x 10(-11) M as compared with 1 x 10(-9) M for the OX7-momordin immunotoxins. Neither of the immunotoxins was toxic to mouse lymphoma EL4 cells, which lack the Thy 1.1 antigen, at concentrations up to 3 x 10(-8) M. Further, bryodin and momordin immunotoxins made from an antibody (R10) of irrelevant specificity were without effect on AKR-A cells. PMID:3265330

  8. [Analysis on intestinal disorders in Jiujing Tu (Illustration of Moxiustion) found from Dunhuang].

    PubMed

    Shi, Yin; Shang, Haixia; Wu, Huangan

    2016-05-01

    Jiujing Tu (Illustration of Moxibustion), excavated from Mo Kao Grotto at Dunhuang, is one of the earliest existing monographs on moxibustion. The medical masters from different schools have focused on this book because it is different from the existing ancient medical works and have not been collected in the medical works of different dynasties. In this study, the literature of Jiujing Tu on five acupoints (Dachangshu, Pangguangshu, Daxiaochangshu, Nieshu and Cigong) relevant with intestinal disorders is collected. It is intended to discuss and analyze the acupoint location, main intestinal disorder, moxibustion characters, recognition on the literature of different dynasties and modern clinical applications. It is believed that the thought of strong moxibustion in the treatment of intestinal disorders advocated in Jiujing Tu has profound impact on the medical development in later generations. It deserves us to have a further digging, collection and promotion of this thought in the modern time. PMID:27509625

  9. Thy1-hAPPLond/Swe+ mouse model of Alzheimer's disease displays broad behavioral deficits in sensorimotor, cognitive and social function

    PubMed Central

    Faizi, Mehrdad; Bader, Patrick L; Saw, Nay; Nguyen, Thuy-Vi V; Beraki, Simret; Wyss-Coray, Tony; Longo, Frank M; Shamloo, Mehrdad

    2012-01-01

    Alzheimer's disease (AD), the most common form of dementia, is an age-dependent progressive neurodegenerative disorder. β-amyloid, a metabolic product of the amyloid precursor protein (APP), plays an important role in the pathogenesis of AD. The Thy1-hAPPLond/Swe+ (line 41) transgenic mouse overexpresses human APP751 and contains the London (V717I) and Swedish (K670M/N671L) mutations. Here, we used a battery of behavioral tests to evaluate general activity, cognition, and social behavior in six-month-old male Thy1-hAPPLond/Swe+ mice. We found hyperactivity in a novel environment as well as significant deficits in spontaneous alternation behavior. In fear conditioning (FC), Thy1-hAPPLond/Swe+ mice did not display deficits in acquisition or in memory retrieval in novel context of tone-cued FC, but they showed significant memory retrieval impairment during contextual testing in an identical environment. Surprisingly, in a standard hidden platform water maze, no significant deficit was detected in mutant mice. However, a delayed-matching-to-place paradigm revealed a significant deficit in Thy1-hAPPLond/Swe+ mice. Lastly, in the social novelty session of a three-chamber test, Thy1-hAPPLond/Swe+ mice exhibited a significantly decreased interest in a novel versus a familiar stranger compared to control mice. This could possibly be explained by decreased social memory or discrimination and may parallel disturbances in social functioning in human AD patients. In conclusion, the Thy1-hAPPLond/Swe+ mouse model of AD displayed a behavioral phenotype that resembles, in part, the cognitive and psychiatric symptoms experienced in AD patients. PMID:22574282

  10. Effects of the color additive caramel color III and 2-acetyl-4(5)-tetrahydroxybutylimidazole (THI) on the immune system of rats.

    PubMed

    Houben, G F; van den Berg, H; Kuijpers, M H; Lam, B W; van Loveren, H; Seinen, W; Penninks, A H

    1992-03-01

    Administration of ammonia caramel color (AC) to rats may decrease blood lymphocyte counts, specifically in rats fed a diet low in vitamin B6. This effect is associated with 2-acetyl-4(5)-(1,2,3,4-tetrahydroxybutyl)imidazole (THI). To characterize and compare the effects of AC and THI and to study the influence of dietary pyridoxine, two studies in rats were conducted. Weanling rats fed a diet containing 2-3 ppm pyridoxine and exposed to 4% AC or 5.72 ppm THI in drinking water for 4 weeks showed reduced cell numbers in spleen and popliteal lymph nodes, as well as in the blood. Flow cytometric analyses demonstrated a comparable reduction in B and T lymphocytes. In blood, spleen, and popliteal lymph nodes, CD4+ lymphocytes were more reduced than CD8+ cells. The number of bone marrow cells was not affected. Although thymus weight and cell number were not affected either, a decreased cortex over medulla area ratio and an increase in medullary cell density largely due to an increase in CD4+ thymocytes was observed. Decreased numbers of ED2+ macrophages were observed in the thymic cortex and in the spleen red pulp. All effects observed were either less pronounced or absent in a study with rats fed a diet containing 11-12 ppm pyridoxine. The effects of AC and THI on the immune system were similar. Whereas AC exposure was associated with changes in vitamin B6 status, THI did not induce obvious effects on vitamin B6 parameters. It is proposed that the effects of AC and THI on the immune system are not caused by a disturbance of vitamin B6 metabolism, but may in fact result from a disturbance of a specific PLP-dependent process. PMID:1553755

  11. A Sex Chromosomal Restriction-Fragment-Length Marker Linked to Melanoma-Determining Tu Loci in Xiphophorus

    PubMed Central

    Schartl, M.

    1988-01-01

    In Xiphophorus, the causative genetic information for melanoma formation has been assigned by classical genetics to chromosomal loci, which are located on the sex chromosomes. In our attempts to molecularly clone these melanoma-determining loci, named Tu, we have looked for restriction-fragment-length markers (RFLMs) linked to the Tu loci. These RFLMs should be useful in obtaining a physical map of a Tu locus, which will aid in the cloning of the corresponding sequences. DNA samples from various Xiphophorus strains and hybrids including those bearing different Tu wild-type, deletion and translocation chromosomes, were screened for the presence of random RFLMs using homologous or heterologous sequences as hybridization probes. We find an EcoRI restriction fragment which shows limited crosshybridization to the v-erb B gene--but not representing the authentic c-erb B gene of Xiphophorus--to be polymorphic with respect to different sex chromosomes. Linkage analysis revealed that a 5-kb fragment is linked to the Tu-Sd locus on the X chromosome, a 7-kb fragment is linked to the Tu-Sr locus on the Y chromosome, both of Xiphophorus maculatus, and that a 12-kb fragment is linked to the Tu-Li locus on the X chromosome of Xiphophorus variatus. Using different chromosomal mutants this RFLM has been mapped to a frequent deletion/translocation breakpoint of the X chromosome, less than 0.3 cM apart from the Tu locus. PMID:2841190

  12. A sex chromosomal restriction-fragment-length marker linked to melanoma-determining Tu loci in Xiphophorus.

    PubMed

    Schartl, M

    1988-07-01

    In Xiphophorus, the causative genetic information for melanoma formation has been assigned by classical genetics to chromosomal loci, which are located on the sex chromosomes. In our attempts to molecularly clone these melanoma-determining loci, named Tu, we have looked for restriction-fragment-length markers (RFLMs) linked to the Tu loci. These RFLMs should be useful in obtaining a physical map of a Tu locus, which will aid in the cloning of the corresponding sequences. DNA samples from various Xiphophorus strains and hybrids including those bearing different Tu wild-type, deletion and translocation chromosomes, were screened for the presence of random RFLMs using homologous or heterologous sequences as hybridization probes. We find an EcoRI restriction fragment which shows limited crosshybridization to the v-erb B gene--but not representing the authentic c-erb B gene of Xiphophorus--to be polymorphic with respect to different sex chromosomes. Linkage analysis revealed that a 5-kb fragment is linked to the Tu-Sd locus on the X chromosome, a 7-kb fragment is linked to the Tu-Sr locus on the Y chromosome, both of Xiphophorus maculatus, and that a 12-kb fragment is linked to the Tu-Li locus on the X chromosome of Xiphophorus variatus. Using different chromosomal mutants this RFLM has been mapped to a frequent deletion/translocation breakpoint of the X chromosome, less than 0.3 cM apart from the Tu locus. PMID:2841190

  13. Identification and cloning of two immunogenic Clostridium perfringens proteins, elongation factor Tu and pyruvate:ferredoxin oxidoreductase of C. perfringens

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Clostridium-related poultry diseases such as necrotic enteritis (NE) and gangrenous dermatitis (GD) cause substantial economic losses on a global scale. Two antigenic Clostridium perfringens proteins, elongation factor Tu (EF-Tu) and pyruvate:ferredoxin oxidoreductase (PFO), were identified by react...

  14. Heat tolerance and expression of protein synthesis elongation factors, EF-Tu and EF-1a, in spring wheat

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Protein elongation factors, EF-Tu and EF-1a, have been implicated in cell response to heat stress. In spring wheat, EF-Tu displays chaperone activity and reduces thermal aggregation of Rubisco activase. Similarly, in mammalian cells, EF-1a displays chaperone-like activity and regulates the expressio...

  15. Do maise and wheat chloroplast protein synthesis elongation factor, EF-Tu, protect Rubisco activase from thermal aggregation and inactivation?

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Maize (Zea mays L.) chloroplast protein synthesis elongation factor, EF-Tu, has been implicated in the development of heat tolerance. The precursor of this protein (pre-EF-Tu) has been shown to display chaperone activity, as it protected heat labile citrate synthase and malate dehydrogenase from the...

  16. Youyou Tu: significance of winning the 2015 Nobel Prize in Physiology or Medicine.

    PubMed

    Liu, Wenxiu; Liu, Yue

    2016-02-01

    Youyou Tu, a female scientist at the China Academy of Traditional Chinese Medicine in Beijing, is the first Chinese winner of the Nobel Prize in Physiology or Medicine. Based on the study of recipes which had been used for thousands of years to treat fever, Tu's group discovered that the plant artemesia annua, sweet wormwood, showed substantial inhibition of rodent malaria parasites. Her achievement and experience have inspired other researchers and emphasized the development of traditional Chinese medicine. Her award has led to a heated discussion about scientific research investment, fair treatment of research staff, and intellectual property right (IPR) protection in China. PMID:26885485

  17. Carbonyl sulfide hydrolase from Thiobacillus thioparus strain THI115 is one of the β-carbonic anhydrase family enzymes.

    PubMed

    Ogawa, Takahiro; Noguchi, Keiichi; Saito, Masahiko; Nagahata, Yoshiko; Kato, Hiromi; Ohtaki, Akashi; Nakayama, Hiroshi; Dohmae, Naoshi; Matsushita, Yasuhiko; Odaka, Masafumi; Yohda, Masafumi; Nyunoya, Hiroshi; Katayama, Yoko

    2013-03-13

    Carbonyl sulfide (COS) is an atmospheric trace gas leading to sulfate aerosol formation, thereby participating in the global radiation balance and ozone chemistry, but its biological sinks are not well understood. Thiobacillus thioparus strain THI115 can grow on thiocyanate (SCN(-)) as its sole energy source. Previously, we showed that SCN(-) is first converted to COS by thiocyanate hydrolase in T. thioparus strain THI115. In the present work, we purified, characterized, and determined the crystal structure of carbonyl sulfide hydrolase (COSase), which is responsible for the degradation of COS to H2S and CO2, the second step of SCN(-) assimilation. COSase is a homotetramer composed of a 23.4 kDa subunit containing a zinc ion in its catalytic site. The amino acid sequence of COSase is homologous to the β-class carbonic anhydrases (β-CAs). Although the crystal structure including the catalytic site resembles those of the β-CAs, CO2 hydration activity of COSase is negligible compared to those of the β-CAs. The α5 helix and the extra loop (Gly150-Pro158) near the N-terminus of the α6 helix narrow the substrate pathway, which could be responsible for the substrate specificity. The k(cat)/K(m) value, 9.6 × 10(5) s(-1) M(-1), is comparable to those of the β-CAs. COSase hydrolyzes COS over a wide concentration range, including the ambient level, in vitro and in vivo. COSase and its structurally related enzymes are distributed in the clade D in the phylogenetic tree of β-CAs, suggesting that COSase and its related enzymes are one of the catalysts responsible for the global sink of COS. PMID:23406161

  18. EF-Tu from the enacyloxin producing Frateuria W-315 strain: Structure/activity relationship and antibiotic resistance.

    PubMed

    Créchet, Jean-Bernard; Malosse, Christian; Hountondji, Codjo

    2016-08-01

    In this report, we have demonstrated that the poly(U)-dependent poly(Phe) synthesis activity of elongator factor Tu (EF-Tu) from the enacyloxin producing strain Frateuria sp. W-315 is inhibited by the antibiotic similarly to that of Escherichia coli EF-Tu. The inhibitory effect of enacyloxin observed in a purified system was the same as that obtained with an S30 extract from E. coli or Frateuria sp. W-315, respectively, suggesting that antibiotic resistance of enacyloxin producing Frateuria sp. W-315 is not due neither to EF-Tu nor to other components of the translation machinery but to a still unknown mechanism. The EF-Tu gene, as PCR amplified from Frateuria W-315 genomic DNA and sequenced represented an ORF of 1191 nucleotides corresponding to 396 amino acids. This protein is larger than the product of tufA from E. coli by only two amino acid residues. Alignment of the amino acid sequence of EF-Tu from E. coli with those of Frateuria and Ralstonia solanacearum indicates on average 80% identical amino acid residues and 9.7% conservative replacements between EF-Tu Frateuria and EF-Tu E. coli, on one hand, and 97% identity and 1.7% conservative replacement between EF-Tu Frateuria and EF-Tu Ralstonia solanacearum, on the other hand. These strong primary structure similarities between EF-Tu from different origins are consistent with the fact that this factor is essential for the translation process in all kingdoms of life. Comparison of the effects of antibiotics on EF-Tu Frateuria and EF-Tu E. coli revealed that enacyloxin, kirromycin and pulvomycin exert a stronger stimulation of the GDP dissociation rate on EF-Tu Frateuria, while the effects of the antibiotics on the GDP association rate were comparable for the two EF-Tu species. Different mutants of EF-Tu E. coli were constructed with the help of site directed mutagenesis by changing one or several residues of EF-Tu E. coli by the corresponding residues of EF-Tu Frateuria. The single A45K substitution did

  19. Identification of a novel bone marrow-derived B-cell progenitor population that coexpresses B220 and Thy-1 and is highly enriched for Abelson leukemia virus targets.

    PubMed Central

    Tidmarsh, G F; Heimfeld, S; Whitlock, C A; Weissman, I L; Müller-Sieburg, C E

    1989-01-01

    A novel stage in early B-lymphocyte differentiation has been identified in normal mouse bone marrow cells. Earlier work had demonstrated that bone marrow cells characterized by low levels of Thy-1 and lack of a panel of lineage markers (Thy-1lo Lin- cells) were highly enriched for pluripotent hematopoietic stem cells. In this paper, we present evidence that another bone marrow population, which expressed low levels of Thy-1 and coexpressed B220, a B-lineage-specific form of the leukocyte common antigen, contained early and potent precursors for B lymphocytes upon in vivo transfer to irradiated hosts. These Thy-1lo B220+ cells, comprising 1 to 2% of bone marrow cells, were enriched for large cells in the mitotic cycle; the population lacked significant pluripotent hematopoietic stem cell activity and myeloid-erythroid progenitors. Most strikingly, Thy-1lo B220+ cells represented a highly enriched population of bone marrow cells that could be targets of Abelson murine leukemia virus transformation. We propose that Thy-1lo B220+ bone marrow cells represent the earliest stage of committed lymphocyte progenitors, intermediate in differentiation between Thy-1lo Lin- pluripotent stem cells and, in the B lineage, Thy-1- B220+ pre-B cells. Images PMID:2474759

  20. Synthesis and application of (13)C-labeled 2-acetyl-4-((1R,2S,3R)-1,2,3,4-tetrahydroxybutyl)imidazole (THI), an immunosuppressant observed in caramel food colorings.

    PubMed

    Elsinghorst, Paul W; Raters, Marion; Dingel, Anna; Fischer, Jochen; Matissek, Reinhard

    2013-08-01

    2-Acetyl-4-((1R,2S,3R)-1,2,3,4-tetrahydroxybutyl)imidazole (THI) is a minor toxic contaminant observed in caramel food colorings and was shown to exert immunosuppressant activity when fed to rodents. Because of this toxicity, maximum levels of THI in caramel food colorings have been defined by international and European authorities. Several reports of THI analysis using external standardization have been published for liquid foods such as beers and soft drinks. However, no suitable internal standard has yet been described allowing THI analysis in more complex samples. In this paper we describe the preparation of a labeled [(13)C6]THI analogue and its application for the successful validation of the first stable isotope dilution assay (SIDA) of THI in caramel food colorings. A brief survey of THI levels in commercially available caramel class III (E 150c) and IV (E 150d) food colorings is also included, corroborating that THI occurs only in caramel class III food colorings. PMID:23866086

  1. “Impact of CB6 and CB05TU chemical mechanisms on air quality”

    EPA Science Inventory

    “Impacts of CB6 and CB05TU chemical mechanisms on air quality”In this study, we incorporate the newly developed Carbon Bond chemical mechanism (CB6) into the Community Multiscale Air Quality modeling system (CMAQv5.0.1) and perform air quality model simulations with the CB6 and t...

  2. Pyridoxine supplementation protects mice from suppression of contact hypersensitivity induced by 2-acetyl-4-tetrahydroxybutylimidazole (THI), ultraviolet B radiation (280-320 nm), or cis-urocanic acid.

    PubMed

    Reeve, V E; Bosnic, M; Boehm-Wilcox, C; Cope, R B

    1995-03-01

    Evidence exists implicating the epidermal ultraviolet B (UVB) photoproduct cis-urocanic acid as an immunogenic mediator of the systemic suppression of T cell-mediated immunity by UVB exposure. Cis-urocanic acid appears to act via histamine receptor pathways, and histamine receptor antagonists and other imidazole ring compounds may modify its immune suppressing action. A component of the food coloring substance ammonia caramel, 2-acetyl-4-tetrahydroxybutylimidazole (THI), which is known to cause lymphopenia in rats, appears to suppress immunity by a similar pathway when the contact hypersensitivity reaction has been the immune function assay in mice. The induction of lymphopenia in rats by THI is inhibited by the vitamin pyridoxine. This study demonstrates that the suppression of contact hypersensitivity in mice by UVB radiation, cis-urocanic acid, or THI is strongly inhibited by supplemental pyridoxine, fed at 30 mg/kg diet, in comparison with the normal diet, which supplies 7 mg pyridoxine/kg diet. These results suggest that pyridoxine competes with cis-urocanic acid and THI for the same binding site or receptor, which we postulate to be a histamine-like T lymphocyte receptor, and that a role may exist for the control of photoimmunosuppression by this vitamin. PMID:7872221

  3. Analyses of acute graft-versus-host-like reaction in (MRL/lpr----MRL/+) chimeric mice using MRL/lpr-Thy-1. 1 congenic mice

    SciTech Connect

    Nakagawa, T.; Nagata, N.; Hosaka, N.; Inaba, M.; Yasumizu, R.; Ogawa, R.; Ikehara, S. )

    1991-10-01

    When MRL/Mp(-)+/+(MRL/+) mice are lethally irradiated and then reconstituted with MRL/Mp-lpr/lpr (MRL/lpr) bone marrow and/or spleen cells, these MRL/+ mice develop lpr-GVHD which is similar to acute graft-versus-host disease (GVHD). Using a Thy-1 congenic strain of MRL/lpr mice (MRL/lpr-Thy-1.1), the authors analyzed T cell subpopulations in the thymus and spleen of MRL/+ mice suffering from lpr-GVHD. lpr-GVHD was induced in MRL/+ mice by transplantation of bone marrow cells (BMC) from MRL/lpr-Thy-1.1 mice; severe lymphocyte depletion associated with fibrosis was observed in the spleens after 7 weeks of bone marrow transplantation (BMT). Thymocytes of the host MRL/+ thymus were replaced with donor-derived cells from the early stage of lpr-GVHD, whereas in the spleen, a small number of host T cells (Thy-1.2+) (4-5%) were retained until the late stage of lpr-GVHD. Donor-type (Thy-1.1+) T cell subsets were not different from those of nontreated MRL/+ mice in the thymus, whereas in the spleen. CD8+ T cells (Thy-1.1+) reached a peak at 5 weeks after BMT, and CD4+ T cells (Thy-1.1+), a peak at 6 weeks. The elimination of T cells from MRL/lpr BMC had no evident effect on the prevention of lpr-GVHD. T cell subpopulations showed a similar pattern to GVHD elicited by MHC differences. Analyses of autoreactive T cells expressing V beta 5 or V beta 11 revealed that autoreactive T cells were deleted from the peripheral lymph nodes. Interestingly, the levels of IgG anti-ssDNA antibodies markedly increased, and both IgM and IgG rheumatoid factors slightly increased 5 to 7 weeks after BMT. These findings are discussed in relation to not only GVHD elicited by MHC differences but also autoimmune diseases.

  4. Human oxygen sensing may have origins in prokaryotic elongation factor Tu prolyl-hydroxylation

    PubMed Central

    Scotti, John S.; Leung, Ivanhoe K. H.; Ge, Wei; Bentley, Michael A.; Paps, Jordi; Kramer, Holger B.; Lee, Joongoo; Aik, WeiShen; Choi, Hwanho; Paulsen, Steinar M.; Bowman, Lesley A. H.; Loik, Nikita D.; Horita, Shoichiro; Ho, Chia-hua; Kershaw, Nadia J.; Tang, Christoph M.; Claridge, Timothy D. W.; Preston, Gail M.; McDonough, Michael A.; Schofield, Christopher J.

    2014-01-01

    The roles of 2-oxoglutarate (2OG)-dependent prolyl-hydroxylases in eukaryotes include collagen stabilization, hypoxia sensing, and translational regulation. The hypoxia-inducible factor (HIF) sensing system is conserved in animals, but not in other organisms. However, bioinformatics imply that 2OG-dependent prolyl-hydroxylases (PHDs) homologous to those acting as sensing components for the HIF system in animals occur in prokaryotes. We report cellular, biochemical, and crystallographic analyses revealing that Pseudomonas prolyl-hydroxylase domain containing protein (PPHD) contain a 2OG oxygenase related in structure and function to the animal PHDs. A Pseudomonas aeruginosa PPHD knockout mutant displays impaired growth in the presence of iron chelators and increased production of the virulence factor pyocyanin. We identify elongation factor Tu (EF-Tu) as a PPHD substrate, which undergoes prolyl-4-hydroxylation on its switch I loop. A crystal structure of PPHD reveals striking similarity to human PHD2 and a Chlamydomonas reinhardtii prolyl-4-hydroxylase. A crystal structure of PPHD complexed with intact EF-Tu reveals that major conformational changes occur in both PPHD and EF-Tu, including a >20-Å movement of the EF-Tu switch I loop. Comparison of the PPHD structures with those of HIF and collagen PHDs reveals conservation in substrate recognition despite diverse biological roles and origins. The observed changes will be useful in designing new types of 2OG oxygenase inhibitors based on various conformational states, rather than active site iron chelators, which make up most reported 2OG oxygenase inhibitors. Structurally informed phylogenetic analyses suggest that the role of prolyl-hydroxylation in human hypoxia sensing has ancient origins. PMID:25197067

  5. Structural elements defining elongation factor Tu mediated suppression of codon ambiguity

    PubMed Central

    Roy, Hervé; Becker, Hubert Dominique; Mazauric, Marie-Hélène; Kern, Daniel

    2007-01-01

    In most prokaryotes Asn-tRNAAsn and Gln-tRNAGln are formed by amidation of aspartate and glutamate mischarged onto tRNAAsn and tRNAGln, respectively. Coexistence in the organism of mischarged Asp-tRNAAsn and Glu-tRNAGln and the homologous Asn-tRNAAsn and Gln-tRNAGln does not, however, lead to erroneous incorporation of Asp and Glu into proteins, since EF-Tu discriminates the misacylated tRNAs from the correctly charged ones. This property contrasts with the canonical function of EF-Tu, which is to non-specifically bind the homologous aa-tRNAs, as well as heterologous species formed in vitro by aminoacylation of non-cognate tRNAs. In Thermus thermophilus that forms the Asp-tRNAAsn intermediate by the indirect pathway of tRNA asparaginylation, EF-Tu must discriminate the mischarged aminoacyl-tRNAs (aa-tRNA). We show that two base pairs in the tRNA T-arm and a single residue in the amino acid binding pocket of EF-Tu promote discrimination of Asp-tRNAAsn from Asn-tRNAAsn and Asp-tRNAAsp by the protein. Our analysis suggests that these structural elements might also contribute to rejection of other mischarged aa-tRNAs formed in vivo that are not involved in peptide elongation. Additionally, these structural features might be involved in maintaining a delicate balance of weak and strong binding affinities between EF-Tu and the amino acid and tRNA moieties of other elongator aa-tRNAs. PMID:17478519

  6. Design and properties of efficient tRNA:EF-Tu FRET system for studies of ribosomal translation

    PubMed Central

    Chudaev, Maxim; Poruri, Kiran; Goldman, Emanuel; Jakubowski, Hieronim; Jain, Mohit Raja; Chen, Wei; Li, Hong; Tyagi, Sanjay; Mandecki, Wlodek

    2013-01-01

    Formation of the ternary complex between GTP-bound form of elongation factor Tu (EF-Tu) and aminoacylated transfer RNA (aa-tRNA) is a key event in protein biosynthesis. Here we show that fluorescently modified Escherichia coli EF-Tu carrying three mutations, C137A, C255V and E348C, and fluorescently modified Phe-tRNAPhe form functionally active ternary complex that has properties similar to those of the naturally occurring (unmodified) complex. Similarities include the binding and binding rate constants, behavior in gel retardation assay, as well as activities in tRNA protection and in vitro translation assays. Proper labeling of EF-Tu was demonstrated in MALDI mass spectroscopy experiments. To generate the mutant EF-Tu, a series of genetic constructions were performed. Two native cysteine residues in the wild-type EF-Tu at positions 137 and 255 were replaced by Ala and Val, respectively, and an additional cysteine was introduced either in position 324 or 348. The assembly FRET assay showed a 5- to 7-fold increase of Cy5-labeled EF-Tu E348C mutant fluorescence upon formation of ternary complex with charged tRNAPhe(Cy3-labeled) when the complex was excited at 532 nm and monitored at 665 nm. In a control experiment, we did not observe FRET using uncharged tRNAPhe(Cy3), nor with wild-type EF-Tu preparation that was allowed to react with Cy5 maleimide, nor in the absence of GTP. The results obtained demonstrate that the EF-Tu:tRNA FRET system described can be used for investigations of ribosomal translation in many types of experiments. PMID:23447652

  7. Inactivation of thyA in Staphylococcus aureus attenuates virulence and has a strong impact on metabolism and virulence gene expression.

    PubMed

    Kriegeskorte, Andre; Block, Desiree; Drescher, Mike; Windmüller, Nadine; Mellmann, Alexander; Baum, Cathrin; Neumann, Claudia; Lorè, Nicola Ivan; Bragonzi, Alessandra; Liebau, Eva; Hertel, Patrick; Seggewiss, Jochen; Becker, Karsten; Proctor, Richard A; Peters, Georg; Kahl, Barbara C

    2014-01-01

    Staphylococcus aureus thymidine-dependent small-colony variants (TD-SCVs) are frequently isolated from patients with chronic S. aureus infections after long-term treatment with trimethoprim-sulfamethoxazole (TMP-SMX). While it has been shown that TD-SCVs were associated with mutations in thymidylate synthase (TS; thyA), the impact of such mutations on protein function is lacking. In this study, we showed that mutations in thyA were leading to inactivity of TS proteins, and TS inactivity led to tremendous impact on S. aureus physiology and virulence. Whole DNA microarray analysis of the constructed ΔthyA mutant identified severe alterations compared to the wild type. Important virulence regulators (agr, arlRS, sarA) and major virulence determinants (hla, hlb, sspAB, and geh) were downregulated, while genes important for colonization (fnbA, fnbB, spa, clfB, sdrC, and sdrD) were upregulated. The expression of genes involved in pyrimidine and purine metabolism and nucleotide interconversion changed significantly. NupC was identified as a major nucleoside transporter, which supported growth of the mutant during TMP-SMX exposure by uptake of extracellular thymidine. The ΔthyA mutant was strongly attenuated in virulence models, including a Caenorhabditis elegans killing model and an acute pneumonia mouse model. This study identified inactivation of TS as the molecular basis of clinical TD-SCV and showed that thyA activity has a major role for S. aureus virulence and physiology. Importance: Thymidine-dependent small-colony variants (TD-SCVs) of Staphylococcus aureus carry mutations in the thymidylate synthase (TS) gene (thyA) responsible for de novo synthesis of thymidylate, which is essential for DNA synthesis. TD-SCVs have been isolated from patients treated for long periods with trimethoprim-sulfamethoxazole (TMP-SMX) and are associated with chronic and recurrent infections. In the era of community-associated methicillin-resistant S. aureus, the therapeutic use of TMP

  8. The Heme Oxygenase-1 Inducer THI-56 Negatively Regulates iNOS Expression and HMGB1 Release in LPS-Activated RAW 264.7 Cells and CLP-Induced Septic Mice

    PubMed Central

    Kim, Young Min; Park, Sang Won; Kim, Hye Jung; Lee, Jae Heun; Chang, Ki Churl

    2013-01-01

    The nuclear DNA binding protein high mobility group box 1 (HMGB1) has recently been suggested to act as a late mediator of septic shock. The effect of ((S)-6,7-dihydroxy-1-(4-hydroxynaphthylmethyl)-1,2,3,4-tetrahydroisoquinoline alkaloid, also known as THI-56, in an experimental model of sepsis was investigated. THI-56 exhibited potent anti-inflammatory properties in response to LPS in RAW 264.7 cells. In particular, THI-56 significantly inhibited the expression of inducible nitric oxide synthase (iNOS) and the release of HMGB1 in activated macrophages. THI-56 activated NE-F2-regulated factor 2 (Nrf-2)/heme oxygenase 1 (HO-1). The specific knockdown of the HO-1 gene by HO-1 siRNA significantly reversed the inhibitory effects of THI-56 on iNOS expression and HMGB1 release in LPS-stimulated macrophages. Importantly, THI-56 administration protected animals from death induced by either a lethal dose of LPS or cecal ligation and puncture (CLP). Furthermore, the ALT, AST, BUN, creatinine, and HMGB1 levels in the blood were significantly increased in CLP-induced septic mice, and the administration of THI-56 reduced these levels in a concentration-dependent and zinc protoporphyrin IX (ZnPPIX)-sensitive manner. In addition, the administration of THI-56 significantly ameliorated not only lung damage but also macrophage infiltration in the livers of CLP-induced septic mice, and these effects were also abrogated in the presence of ZnPPIX. Thus, we conclude that THI-56 significantly attenuates the proinflammatory response induced by LPS and reduces organ damage in a CLP-induced sepsis model through the upregulation of Nrf-2/HO-1. PMID:24098466

  9. First insights into the genetic diversity and origin of Leishmania infantum in Mont Rolland (Thiès region, Senegal).

    PubMed

    Cassan, Cecile; Dione, Michel Mainack; Dereure, Jacques; Diedhiou, Souleymane; Bucheton, Bruno; Hide, Mallorie; Kako, Caroline; Gaye, Oumar; Senghor, Massila; Niang, Abdoul Aziz; Bañuls, Anne-Laure; Faye, Babacar

    2016-06-01

    Visceral leishmaniasis is not endemic in West Africa. However, high seroprevalence of Leishmania infantum infection (one of the Leishmania species that cause visceral leishmaniasis) was detected in dogs and humans in the Mont Rolland community (close to Thiès, Senegal), despite the lack of reports concerning human clinical cases. Our aim was to genetically characterize this L. infantum population and identify its origin. We thus conducted seven field surveys in 25 villages of the Mont Rolland community between 2005 and 2009 and blood samples were collected from 205 dogs. Serological testing indicated that 92 dogs (44.9%) were positive for Leishmania infection. L. infantum was identified as the cause of infection. Analysis of 29 L. infantum isolates from these dogs by multilocus microsatellite typing and multilocus sequence typing indicated that this population had very limited genetic diversity, low level of heterozygosity and only seven different genotypes (79.3% of all isolates had the same genotype). Multilocus sequence typing showed that the Mont Rolland isolates clustered with strains from the Mediterranean basin and were separated from East African and Asian strains. Therefore, our data suggest a quite recent and unique introduction into Senegal of a L. infantum strain from the Mediterranean basin. PMID:26945844

  10. CD90- (Thy-1-) High Selection Enhances Reprogramming Capacity of Murine Adipose-Derived Mesenchymal Stem Cells

    PubMed Central

    Kawamoto, Koichi; Konno, Masamitsu; Nagano, Hiroaki; Nishikawa, Shimpei; Tomimaru, Yoshito; Akita, Hirofumi; Hama, Naoki; Wada, Hiroshi; Kobayashi, Shogo; Eguchi, Hidetoshi; Tanemura, Masahiro; Ito, Toshinori; Doki, Yuichiro; Mori, Masaki; Ishii, Hideshi

    2013-01-01

    Background. Mesenchymal stem cells (MSCs), including adipose tissue-derived mesenchymal stem cells (ADSC), are multipotent and can differentiate into various cell types possessing unique immunomodulatory features. Several clinical trials have demonstrated the safety and possible efficacy of MSCs in organ transplantation. Thus, stem cell therapy is promising for tolerance induction. In this study, we assessed the reprogramming capacity of murine ADSCs and found that CD90 (Thy-1), originally discovered as a thymocyte antigen, could be a useful marker for cell therapy. Method. Murine ADSCs were isolated from B6 mice, sorted using a FACSAria cell sorter by selection of CD90Hi or CD90Lo, and then transduced with four standard factors (4F; Oct4, Sox2, Klf4, and c-Myc). Results. Unsorted, CD90Hi-sorted, and CD90Lo-sorted murine ADSCs were reprogrammed using standard 4F transduction. CD90Hi ADSCs showed increased numbers of alkaline phosphatase-positive colonies compared with CD90Lo ADSCs. The relative reprogramming efficiencies of unsorted, CD90Hi-sorted, and CD90Lo-sorted ADSCs were 100%, 116.5%, and 74.7%, respectively. CD90Hi cells were more responsive to reprogramming. Conclusion. CD90Hi ADSCs had greater reprogramming capacity than CD90Lo ADSCs, suggesting that ADSCs have heterogeneous subpopulations. Thus, CD90Hi selection presents an effective strategy to isolate a highly suppressive subpopulation for stem cell-based tolerance induction therapy. PMID:24282338

  11. Ligand-induced folding of the thiM TPP riboswitch investigated by a structure-based fluorescence spectroscopic approach

    PubMed Central

    Lang, Kathrin; Rieder, Renate; Micura, Ronald

    2007-01-01

    Riboswitches are genetic control elements within non-coding regions of mRNA. They consist of a metabolite-sensitive aptamer and an adjoining expression platform. Here, we describe ligand-induced folding of a thiamine pyrophosphate (TPP) responsive riboswitch from Escherichia coli thiM mRNA, using chemically labeled variants. Referring to a recent structure determination of the TPP/aptamer complex, each variant was synthesized with a single 2-aminopurine (AP) nucleobase replacement that was selected to monitor formation of tertiary interactions of a particular region during ligand binding in real time by fluorescence experiments. We have determined the rate constants for conformational adjustment of the individual AP sensors. From the 7-fold differentiation of these constants, it can be deduced that tertiary contacts between the two parallel helical domains (P2/J3-2/P3/L3 and P4/P5/L5) that grip the ligand's ends in two separate pockets, form significantly faster than the function-critical three-way junction with stem P1 fully developed. Based on these data, we characterize the process of ligand binding by an induced fit of the RNA and propose a folding model of the TPP riboswitch aptamer. For the full-length riboswitch domain and for shorter constructs that represent transcriptional intermediates, we have additionally evaluated ligand-induced folding via AP-modified variants and provide insights into the sequential folding pathway that involves a finely balanced equilibrium of secondary structures. PMID:17693433

  12. Oxidation of a Cysteine Residue in Elongation Factor EF-Tu Reversibly Inhibits Translation in the Cyanobacterium Synechocystis sp. PCC 6803.

    PubMed

    Yutthanasirikul, Rayakorn; Nagano, Takanori; Jimbo, Haruhiko; Hihara, Yukako; Kanamori, Takashi; Ueda, Takuya; Haruyama, Takamitsu; Konno, Hiroki; Yoshida, Keisuke; Hisabori, Toru; Nishiyama, Yoshitaka

    2016-03-11

    Translational elongation is susceptible to inactivation by reactive oxygen species (ROS) in the cyanobacterium Synechocystis sp. PCC 6803, and elongation factor G has been identified as a target of oxidation by ROS. In the present study we examined the sensitivity to oxidation by ROS of another elongation factor, EF-Tu. The structure of EF-Tu changes dramatically depending on the bound nucleotide. Therefore, we investigated the sensitivity to oxidation in vitro of GTP- and GDP-bound EF-Tu as well as that of nucleotide-free EF-Tu. Assays of translational activity with a reconstituted translation system from Escherichia coli revealed that GTP-bound and nucleotide-free EF-Tu were sensitive to oxidation by H2O2, whereas GDP-bound EF-Tu was resistant to H2O2. The inactivation of EF-Tu was the result of oxidation of Cys-82, a single cysteine residue, and subsequent formation of both an intermolecular disulfide bond and sulfenic acid. Replacement of Cys-82 with serine rendered EF-Tu resistant to inactivation by H2O2, confirming that Cys-82 was a target of oxidation. Furthermore, oxidized EF-Tu was reduced and reactivated by thioredoxin. Gel-filtration chromatography revealed that some of the oxidized nucleotide-free EF-Tu formed large complexes of >30 molecules. Atomic force microscopy revealed that such large complexes dissociated into several smaller aggregates upon the addition of dithiothreitol. Immunological analysis of the redox state of EF-Tu in vivo showed that levels of oxidized EF-Tu increased under strong light. Thus, resembling elongation factor G, EF-Tu appears to be sensitive to ROS via oxidation of a cysteine residue, and its inactivation might be reversed in a redox-dependent manner. PMID:26786107

  13. The crystal structure of pyrimidine/thiamin biosynthesis precursor-like domain-containing protein CAE31940 from proteobacterium Bordetella bronchiseptica RB50, and evolutionary insight into the NMT1/THI5 family

    PubMed Central

    Bajor, Jacek; Tkaczuk, Karolina L.; Chruszcz, Maksymilian; Chapman, Hutton; Kagan, Olga; Savchenko, Alexei; Minor, Wladek

    2014-01-01

    We report a 2.0 Å structure of the CAE31940 protein, a proteobacterial NMT1/THI5-like domain-containing protein. We also discuss the primary and tertiary structure similarity with its homologs. The highly conserved FGGXMP motif was identified in CAE31940, which corresponds to the GCCCX motif located in the vicinity of the active center characteristic for THi5-like proteins found in yeast. This suggests that the FGGXMP motif may be a unique hallmark of proteobacterial NMT1/THI5-like proteins. PMID:24908050

  14. Interaction of CK1δ with γTuSC ensures proper microtubule assembly and spindle positioning

    PubMed Central

    Peng, Yutian; Moritz, Michelle; Han, Xuemei; Giddings, Thomas H.; Lyon, Andrew; Kollman, Justin; Winey, Mark; Yates, John; Agard, David A.; Drubin, David G.; Barnes, Georjana

    2015-01-01

    Casein kinase 1δ (CK1δ) family members associate with microtubule-organizing centers (MTOCs) from yeast to humans, but their mitotic roles and targets have yet to be identified. We show here that budding yeast CK1δ, Hrr25, is a γ-tubulin small complex (γTuSC) binding factor. Moreover, Hrr25's association with γTuSC depends on its kinase activity and its noncatalytic central domain. Loss of Hrr25 kinase activity resulted in assembly of unusually long cytoplasmic microtubules and defects in spindle positioning, consistent with roles in regulation of γTuSC-mediated microtubule nucleation and the Kar9 spindle-positioning pathway, respectively. Hrr25 directly phosphorylated γTuSC proteins in vivo and in vitro, and this phosphorylation promoted γTuSC integrity and activity. Because CK1δ and γTuSC are highly conserved and present at MTOCs in diverse eukaryotes, similar regulatory mechanisms are expected to apply generally in eukaryotes. PMID:25971801

  15. Heat shock transcription factor δ³² is targeted for degradation via an ubiquitin-like protein ThiS in Escherichia coli.

    PubMed

    Xu, Xibing; Niu, Yulong; Liang, Ke; Wang, Jianmei; Li, Xufeng; Yang, Yi

    2015-04-01

    The posttranslational modification of proteins with ubiquitin and ubiquitin-like proteins (UBLs) plays an important role in eukaryote biology, through which substrate proteins are targeted for degradation by the proteasome. Prokaryotes have been thought to degrade proteins by an ubiquitin independent pathway. Here, we show that ThiS, an ubiquitin-like protein, is covalently attached to δ(32) and at least 27 other proteins, leading to their subsequent degradation by proteases, in a similar manner to the ubiquitin-proteasome system (UPS) in eukaryotes. Molecular biology and biochemical studies confirm that specific lysine sites in δ(32) can be modified by ThiS. The results presented here establish a new model for δ(32) degradation and show that Escherichia coli uses a small-protein modifier to control protein stability. PMID:25721662

  16. Longitudinal Handling Qualities of the Tu-144LL Airplane and Comparisons With Other Large, Supersonic Aircraft

    NASA Technical Reports Server (NTRS)

    Cox, Timothy H.; Marshall, Alisa

    2000-01-01

    Four flights have been conducted using the Tu-144LL supersonic transport aircraft with the dedicated objective of collecting quantitative data and qualitative pilot comments. These data are compared with the following longitudinal flying qualities criteria: Neal-Smith, short-period damping, time delay, control anticipation parameter, phase delay (omega(sp)*T(theta(2))), pitch bandwidth as a function of time delay, and flight path as a function of pitch bandwidth. Determining the applicability of these criteria and gaining insight into the flying qualities of a large, supersonic aircraft are attempted. Where appropriate, YF-12, XB-70, and SR-71 pilot ratings are compared with the Tu-144LL results to aid in the interpretation of the Tu-144LL data and to gain insight into the application of criteria. The data show that approach and landing requirements appear to be applicable to the precision flightpath control required for up-and-away flight of large, supersonic aircraft. The Neal-Smith, control anticipation parameter, and pitch-bandwidth criteria tend to correlate with the pilot comments better than the phase delay criterion, omega(sp)*T(theta(2)). The data indicate that the detrimental flying qualities implication of decoupled pitch-attitude and flightpath responses occurring for high-speed flight may be mitigated by requiring the pilot to close the loop on flightpath or vertical speed.

  17. LONG-TERM PHOTOMETRIC ANALYSIS OF THE ACTIVE W UMa-TYPE SYSTEM TU BOOTIS

    SciTech Connect

    Coughlin, Jeffrey L.; Dale, Horace A. III; Williamon, Richard M.

    2008-09-15

    We present multicolor light curves for the W UMa-type eclipsing binary TU Boo for two epochs separated by 22 years. An analysis of the O - C diagram indicates the earlier observations took place right in the middle of a major period change, thus allowing for a unique study on mass transfer and period changes in this W UMa-type system. We compute model fits to our light curves, along with the only other published set, using the Wilson-Devinney program, and find temporally correlated changes in the size of the secondary component with anomalies in the O - C diagram. We investigate the cause of these changes and find support for the existence of rapid, large-scale mass transfer between the components. We postulate that this interaction allows them to maintain nearly equal surface temperatures despite having achieved only marginal contact. We also find support for the evolutionary scenario in which TU Boo has undergone a mass ratio reversal in the past, due to large-scale mass transfer so that what is presently the secondary component of TU Boo is in an advanced evolutionary state, oversized due to a helium-enriched core, with a total system age of {>=}10 Gyr.

  18. Synthesis of 5-[3-(2-aminopyrimidin-4-yl)aminopropyn-1-yl]uracil derivative that recognizes Ade-Thy base pairs in double-stranded DNA.

    PubMed

    Ito, Yu; Masaki, Yoshiaki; Kanamori, Takashi; Ohkubo, Akihiro; Seio, Kohji; Sekine, Mitsuo

    2016-01-01

    5-[3-(2-Aminopyrimidin-4-yl)aminopropyn-1-yl]uracil (Ura(Pyr)) was designed as a new nucleobase to recognize Ade-Thy base pair in double-stranded DNA. We successfully synthesized the dexoynucleoside phosphoramidite having Ura(Pyr) and incorporated it into triplex forming oligonucleotides (TFOs). Melting temperature analysis revealed that introduction of Ura(Pyr) into TFOs could effectively stabilize their triplex structures without loss of base recognition capabilities. PMID:26602276

  19. Thy1+IL-7+ lymphatic endothelial cells in iBALT provide a survival niche for memory T-helper cells in allergic airway inflammation

    PubMed Central

    Shinoda, Kenta; Hirahara, Kiyoshi; Iinuma, Tomohisa; Ichikawa, Tomomi; Suzuki, Akane S.; Sugaya, Kaoru; Tumes, Damon J.; Yamamoto, Heizaburo; Hara, Takahiro; Tani-ichi, Shizue; Ikuta, Koichi; Okamoto, Yoshitaka; Nakayama, Toshinori

    2016-01-01

    Memory CD4+ T helper (Th) cells are central to long-term protection against pathogens, but they can also be pathogenic and drive chronic inflammatory disorders. How these pathogenic memory Th cells are maintained, particularly at sites of local inflammation, remains unclear. We found that ectopic lymphoid-like structures called inducible bronchus-associated lymphoid tissue (iBALT) are formed during chronic allergic inflammation in the lung, and that memory-type pathogenic Th2 (Tpath2) cells capable of driving allergic inflammation are maintained within the iBALT structures. The maintenance of memory Th2 cells within iBALT is supported by Thy1+IL-7–producing lymphatic endothelial cells (LECs). The Thy1+IL-7–producing LECs express IL-33 and T-cell–attracting chemokines CCL21 and CCL19. Moreover, ectopic lymphoid structures consisting of memory CD4+ T cells and IL-7+IL-33+ LECs were found in nasal polyps of patients with eosinophilic chronic rhinosinusitis. Thus, Thy1+IL-7–producing LECs control chronic allergic airway inflammation by providing a survival niche for memory-type Tpath2 cells. PMID:27140620

  20. New analysis of the light time effect in TU Ursae Majoris

    NASA Astrophysics Data System (ADS)

    Liška, J.; Skarka, M.; Mikulášek, Z.; Zejda, M.; Chrastina, M.

    2016-05-01

    Context. Recent statistical studies prove that the percentage of RR Lyrae pulsators that are located in binaries or multiple stellar systems is considerably lower than might be expected. This can be better understood from an in-depth analysis of individual candidates. We investigate in detail the light time effect of the most probable binary candidate TU UMa. This is complicated because the pulsation period shows secular variation. Aims: We model possible light time effect of TU UMa using a new code applied on previously available and newly determined maxima timings to confirm binarity and refine parameters of the orbit of the RRab component in the binary system. The binary hypothesis is also tested using radial velocity measurements. Methods: We used new approach to determine brightness maxima timings based on template fitting. This can also be used on sparse or scattered data. This approach was successfully applied on measurements from different sources. To determine the orbital parameters of the double star TU UMa, we developed a new code to analyse light time effect that also includes secular variation in the pulsation period. Its usability was successfully tested on CL Aur, an eclipsing binary with mass-transfer in a triple system that shows similar changes in the O-C diagram. Since orbital motion would cause systematic shifts in mean radial velocities (dominated by pulsations), we computed and compared our model with centre-of-mass velocities. They were determined using high-quality templates of radial velocity curves of RRab stars. Results: Maxima timings adopted from the GEOS database (168) together with those newly determined from sky surveys and new measurements (85) were used to construct an O-C diagram spanning almost five proposed orbital cycles. This data set is three times larger than data sets used by previous authors. Modelling of the O-C dependence resulted in 23.3-yr orbital period, which translates into a minimum mass of the second component of

  1. Research data from hydrology disciplines are safely stored and published in 3TU.Datacentrum

    NASA Astrophysics Data System (ADS)

    Prinčič, Alenka; Rombouts, Jeroen

    2013-04-01

    Good management of research data from different science disciplines such as hydrology, geodesy and climate research, is crucial for reducing the uncertainties in forecasting and predictions. These research data can be well-managed and safely stored in 3TU.Datacentrum and thus become available for future use by the scientific community. 3TU.Datacentrum (datacentrum.3tu.nl) is a portal for science- and technology research data in the Netherlands. It provides long-term archive and permanent access to research data, it provides tools for citation of research datasets and thus enables linking, sharing and reuse of research data. Versatility of disciplines and thus versatility of the data producers' needs is a challenge of the data center. 3TU.Datacentrum offers solutions - as standardized as possible yet different where required. Standardized solution is the data model: datasets, instruments, locations and time frames are all objects on their own, with own metadata and interconnected through rdf relationships. These relations are also used to generate ORE Resource Maps. The file format NetCDF, which is primarily used for sets of multidimensional arrays with metadata included, enables interoperability. For simple datasets a Fedora repository is used where datasets of any format are allowed including that of a specific instrument. Alongside the NetCDF an xml version (ncml) is stored that can be converted to other formats. For complex datasets an OPeNDAP framework is used, which allows querying within and across datasets. Some examples from hydrological research are presented. 1 - For a complex collection of simple datasets consisting of ongoing hydrological measurements from several project partners, using several instruments, on several locations, and over a long period of time, there was a need for central storage in order to share the data and enable easy combination of data sets. The measurements are not repeatable and are also valuable for multidisciplinary research

  2. New analysis of the light time effect in TU Ursae Majoris

    NASA Astrophysics Data System (ADS)

    Liška, J.; Skarka, M.; Mikulášek, Z.; Zejda, M.; Chrastina, M.

    2016-04-01

    Context. Recent statistical studies prove that the percentage of RR Lyrae pulsators that are located in binaries or multiple stellar systems is considerably lower than might be expected. This can be better understood from an in-depth analysis of individual candidates. We investigate in detail the light time effect of the most probable binary candidate TU UMa. This is complicated because the pulsation period shows secular variation. Aims: We model possible light time effect of TU UMa using a new code applied on previously available and newly determined maxima timings to confirm binarity and refine parameters of the orbit of the RRab component in the binary system. The binary hypothesis is also tested using radial velocity measurements. Methods: We used new approach to determine brightness maxima timings based on template fitting. This can also be used on sparse or scattered data. This approach was successfully applied on measurements from different sources. To determine the orbital parameters of the double star TU UMa, we developed a new code to analyse light time effect that also includes secular variation in the pulsation period. Its usability was successfully tested on CL Aur, an eclipsing binary with mass-transfer in a triple system that shows similar changes in the O-C diagram. Since orbital motion would cause systematic shifts in mean radial velocities (dominated by pulsations), we computed and compared our model with centre-of-mass velocities. They were determined using high-quality templates of radial velocity curves of RRab stars. Results: Maxima timings adopted from the GEOS database (168) together with those newly determined from sky surveys and new measurements (85) were used to construct an O-C diagram spanning almost five proposed orbital cycles. This data set is three times larger than data sets used by previous authors. Modelling of the O-C dependence resulted in 23.3-yr orbital period, which translates into a minimum mass of the second component of

  3. Know Thy Impact

    ERIC Educational Resources Information Center

    Hattie, John

    2012-01-01

    Teachers give a lot of feedback, and not all of it is good. This article discusses how to ensure teachers are giving students powerful feedback they can use. For feedback to be effective, teachers need to clarify the goal of the lesson or activity, ensure that students understand the feedback, and seek feedback from students about the…

  4. Share with thy neighbors

    NASA Astrophysics Data System (ADS)

    Chandra, Surendar; Yu, Xuwen

    2007-01-01

    Peer to peer (P2P) systems are traditionally designed to scale to a large number of nodes. However, we focus on scenarios where the sharing is effected only among neighbors. Localized sharing is particularly attractive in scenarios where wide area network connectivity is undesirable, expensive or unavailable. On the other hand, local neighbors may not offer the wide variety of objects possible in a much larger system. The goal of this paper is to investigate a P2P system that shares contents with its neighbors. We analyze the sharing behavior of Apple iTunes users in an University setting. iTunes restricts the sharing of audio and video objects to peers within the same LAN sub-network. We show that users are already making a significant amount of content available for local sharing. We show that these systems are not appropriate for applications that require access to a specific object. We argue that mechanisms that allow the user to specify classes of interesting objects are better suited for these systems. Mechanisms such as bloom filters can allow each peer to summarize the contents available in the neighborhood, reducing network search overhead. This research can form the basis for future storage systems that utilize the shared storage available in neighbors and build a probabilistic storage for local consumption.

  5. Enhancement of innate immune system in monocot rice by transferring the dicotyledonous elongation factor Tu receptor EFR.

    PubMed

    Lu, Fen; Wang, Huiqin; Wang, Shanzhi; Jiang, Wendi; Shan, Changlin; Li, Bin; Yang, Jun; Zhang, Shiyong; Sun, Wenxian

    2015-07-01

    The elongation factor Tu (EF-Tu) receptor (EFR) in cruciferous plants specifically recognizes the N-terminal acetylated elf18 region of bacterial EF-Tu and thereby activates plant immunity. It has been demonstrated that Arabidopsis EFR confers broad-spectrum bacterial resistance in the EFR transgenic solanaceous plants. Here, the transgenic rice plants (Oryza sativa L. ssp. japonica cv. Zhonghua 17) and cell cultures with constitutive expression of AtEFR were developed to investigate whether AtEFR senses EF-Tu and thus enhances bacterial resistance in the monocot plants. We demonstrated that the Xanthomonas oryzae-derived elf18 peptide induced oxidative burst and mitogen-activated protein kinase activation in the AtEFR transgenic rice cells and plants, respectively. Pathogenesis-related genes, such as OsPBZ1, were upregulated dramatically in transgenic rice plant and cell lines in response to elf18 stimulation. Importantly, pretreatment with elf18 triggered strong resistance to X. oryzae pv. oryzae in the transgenic plants, which was largely dependent on the AtEFR expression level. These plants also exhibited enhanced resistance to rice bacterial brown stripe, but not to rice fungal blast. Collectively, the results indicate that the rice plants with heterologous expression of AtEFR recognize bacterial EF-Tu and exhibit enhanced broad-spectrum bacterial disease resistance and that pattern recognition receptor-mediated immunity may be manipulated across the two plant classes, dicots and monocots. PMID:25358295

  6. Identification of an EF-Tu protein that is periplasm-associated and processed in Neisseria gonorrhoeae.

    PubMed

    Porcella, S F; Belland, R J; Judd, R C

    1996-09-01

    A 44 kDa protein is a dominant component of periplasmic extracts of Neisseria gonorrhoeae. Peptide sequence generated from a cyanogen-bromide-cleaved fragment of this protein indicated sequence homology with elongation factor-Tu (EF-Tu). Polyclonal antiserum was made against the 44 kDa protein purified from periplasm extracts of N. gonorrhoeae. The preabsorbed antiserum was immunoblotted against whole-cell lysates on two-dimensional gels. A 44 kDa protein and a smaller 37 kDa protein were recognized by this antiserum. A N. gonorrhoeae gamma phage DNA library was screened and a clone expressing a 44 kDa protein was identified. The DNA insert in this clone contained several genes homologous to genes contained in the str operon of Escherichia coli. One ORF product with a calculated molecular mass of 43 kDa was highly homologous to the EF-TuA of E. coli. A synthetic peptide antiserum specific for a portion of the C terminus of EF-Tu confirmed that the 37 kDa protein in whole-cell lysates of N. gonorrhoeae was a processed form of EF-Tu. Deletion of the tufA gene homologue in N. gonorrhoeae was attempted but was unsuccessful. PMID:8828215

  7. Research Pilot C. Gordon Fullerton in Cockpit of TU-144LL SST Flying Laboratory

    NASA Technical Reports Server (NTRS)

    1998-01-01

    NASA Research pilot C. Gordon Fullerton sits in cockpit of TU-144LL SST Flying Laboratory. Fullerton was one of two NASA pilots who flew the aircraft as part of a joint high speed research program. NASA teamed with American and Russian aerospace industries for an extended period in a joint international research program featuring the Russian-built Tu-144LL supersonic aircraft. The object of the program was to develop technologies for a proposed future second-generation supersonic airliner to be developed in the 21st Century. The aircraft's initial flight phase began in June 1996 and concluded in February 1998 after 19 research flights. A shorter follow-on program involving seven flights began in September 1998 and concluded in April 1999. All flights were conducted in Russia from Tupolev's facility at the Zhukovsky Air Development Center near Moscow. The centerpiece of the research program was the Tu 144LL, a first-generation Russian supersonic jetliner that was modified by its developer/builder, Tupolev ANTK (aviatsionnyy nauchno-tekhnicheskiy kompleks-roughly, aviation technical complex), into a flying laboratory for supersonic research. Using the Tu-144LL to conduct flight research experiments, researchers compared full-scale supersonic aircraft flight data with results from models in wind tunnels, computer-aided techniques, and other flight tests. The experiments provided unique aerodynamic, structures, acoustics, and operating environment data on supersonic passenger aircraft. Data collected from the research program was being used to develop the technology base for a proposed future American-built supersonic jetliner. Although actual development of such an advanced supersonic transport (SST) is currently on hold, commercial aviation experts estimate that a market for up to 500 such aircraft could develop by the third decade of the 21st Century. The Tu-144LL used in the NASA-sponsored research program was a 'D' model with different engines than were used in

  8. Tu-144LL SST Flying Laboratory Front View with Nose Dropped for Takeoff and Landing

    NASA Technical Reports Server (NTRS)

    1998-01-01

    A front view of the Tupolev Tu-144LL supersonic flying laboratory at the Zhukovsky Air Development Center near Moscow, Russia. The plane's nose droops down for take off and landing and is then raised for high-speed flight. NASA teamed with American and Russian aerospace industries for an extended period in a joint international research program featuring the Russian-built Tu-144LL supersonic aircraft. The object of the program was to develop technologies for a proposed future second-generation supersonic airliner to be developed in the 21st Century. The aircraft's initial flight phase began in June 1996 and concluded in February 1998 after 19 research flights. A shorter follow-on program involving seven flights began in September 1998 and concluded in April 1999. All flights were conducted in Russia from Tupolev's facility at the Zhukovsky Air Development Center near Moscow. The centerpiece of the research program was the Tu 144LL, a first-generation Russian supersonic jetliner that was modified by its developer/builder, Tupolev ANTK (aviatsionnyy nauchno-tekhnicheskiy kompleks-roughly, aviation technical complex), into a flying laboratory for supersonic research. Using the Tu-144LL to conduct flight research experiments, researchers compared full-scale supersonic aircraft flight data with results from models in wind tunnels, computer-aided techniques, and other flight tests. The experiments provided unique aerodynamic, structures, acoustics, and operating environment data on supersonic passenger aircraft. Data collected from the research program was being used to develop the technology base for a proposed future American-built supersonic jetliner. Although actual development of such an advanced supersonic transport (SST) is currently on hold, commercial aviation experts estimate that a market for up to 500 such aircraft could develop by the third decade of the 21st Century. The Tu-144LL used in the NASA-sponsored research program was a 'D' model with different

  9. Tu-144LL SST Flying Laboratory Lifts off Runway on a High-Speed Research Flight

    NASA Technical Reports Server (NTRS)

    1998-01-01

    The Tupolev Tu-144LL lifts off from the Zhukovsky Air Development Center near Moscow, Russia, on a 1998 test flight. NASA teamed with American and Russian aerospace industries for an extended period in a joint international research program featuring the Russian-built Tu-144LL supersonic aircraft. The object of the program was to develop technologies for a proposed future second-generation supersonic airliner to be developed in the 21st Century. The aircraft's initial flight phase began in June 1996 and concluded in February 1998 after 19 research flights. A shorter follow-on program involving seven flights began in September 1998 and concluded in April 1999. All flights were conducted in Russia from Tupolev's facility at the Zhukovsky Air Development Center near Moscow. The centerpiece of the research program was the Tu 144LL, a first-generation Russian supersonic jetliner that was modified by its developer/builder, Tupolev ANTK (aviatsionnyy nauchno-tekhnicheskiy kompleks-roughly, aviation technical complex), into a flying laboratory for supersonic research. Using the Tu-144LL to conduct flight research experiments, researchers compared full-scale supersonic aircraft flight data with results from models in wind tunnels, computer-aided techniques, and other flight tests. The experiments provided unique aerodynamic, structures, acoustics, and operating environment data on supersonic passenger aircraft. Data collected from the research program was being used to develop the technology base for a proposed future American-built supersonic jetliner. Although actual development of such an advanced supersonic transport (SST) is currently on hold, commercial aviation experts estimate that a market for up to 500 such aircraft could develop by the third decade of the 21st Century. The Tu-144LL used in the NASA-sponsored research program was a 'D' model with different engines than were used in production-model aircraft. Fifty experiments were proposed for the program and

  10. Russian Tu-144LL SST Roll-out for Joint NASA Research Program

    NASA Technical Reports Server (NTRS)

    1996-01-01

    U.S. Ambassador Pickering addresses Russian and American dignitaries, industry representatives and members of the press during a roll-out ceremony for the modified Tu-144LL supersonic flying laboratory. The ceremony was held at the Zhukovsky Air Development Center near Moscow, Russia, on March 17, 1996. The 'LL' designation for the aircraft stands for Letayuschaya Laboratoriya, which means Flying Laboratory in Russian. NASA teamed with American and Russian aerospace industries for an extended period in a joint international research program featuring the Russian-built Tu-144LL supersonic aircraft. The object of the program was to develop technologies for a proposed future second-generation supersonic airliner to be developed in the 21st Century. The aircraft's initial flight phase began in June 1996 and concluded in February 1998 after 19 research flights. A shorter follow-on program involving seven flights began in September 1998 and concluded in April 1999. All flights were conducted in Russia from Tupolev's facility at the Zhukovsky Air Development Center near Moscow. The centerpiece of the research program was the Tu 144LL, a first-generation Russian supersonic jetliner that was modified by its developer/builder, Tupolev ANTK (aviatsionnyy nauchno-tekhnicheskiy kompleks-roughly, aviation technical complex), into a flying laboratory for supersonic research. Using the Tu-144LL to conduct flight research experiments, researchers compared full-scale supersonic aircraft flight data with results from models in wind tunnels, computer-aided techniques, and other flight tests. The experiments provided unique aerodynamic, structures, acoustics, and operating environment data on supersonic passenger aircraft. Data collected from the research program was being used to develop the technology base for a proposed future American-built supersonic jetliner. Although actual development of such an advanced supersonic transport (SST) is currently on hold, commercial aviation

  11. Russian Tu-144LL SST Roll-Out for Joint NASA Research Program

    NASA Technical Reports Server (NTRS)

    1996-01-01

    The modified Tu-144LL supersonic flying laboratory is rolled out of its hangar at the Zhukovsky Air Development Center near Moscow, Russia in March 1996 at the beginning of a joint U.S. - Russian high-speed flight research program. The 'LL' stands for Letayuschaya Laboratoriya, which means Flying Laboratory. NASA teamed with American and Russian aerospace industries for an extended period in a joint international research program featuring the Russian-built Tu-144LL supersonic aircraft. The object of the program was to develop technologies for a proposed future second-generation supersonic airliner to be developed in the 21st Century. The aircraft's initial flight phase began in June 1996 and concluded in February 1998 after 19 research flights. A shorter follow-on program involving seven flights began in September 1998 and concluded in April 1999. All flights were conducted in Russia from Tupolev's facility at the Zhukovsky Air Development Center near Moscow. The centerpiece of the research program was the Tu 144LL, a first-generation Russian supersonic jetliner that was modified by its developer/builder, Tupolev ANTK (aviatsionnyy nauchno-tekhnicheskiy kompleks-roughly, aviation technical complex), into a flying laboratory for supersonic research. Using the Tu-144LL to conduct flight research experiments, researchers compared full-scale supersonic aircraft flight data with results from models in wind tunnels, computer-aided techniques, and other flight tests. The experiments provided unique aerodynamic, structures, acoustics, and operating environment data on supersonic passenger aircraft. Data collected from the research program was being used to develop the technology base for a proposed future American-built supersonic jetliner. Although actual development of such an advanced supersonic transport (SST) is currently on hold, commercial aviation experts estimate that a market for up to 500 such aircraft could develop by the third decade of the 21st Century. The

  12. Tu-144LL SST Flying Laboratory on Taxiway at Zhukovsky Air Development Center near Moscow, Russia

    NASA Technical Reports Server (NTRS)

    1998-01-01

    The sleek lines of the Tupolev Tu-144LL are evident as it sits on the taxiway at the Zhukovsky Air Development Center near Moscow, Russia. NASA teamed with American and Russian aerospace industries for an extended period in a joint international research program featuring the Russian-built Tu-144LL supersonic aircraft. The object of the program was to develop technologies for a proposed future second-generation supersonic airliner to be developed in the 21st Century. The aircraft's initial flight phase began in June 1996 and concluded in February 1998 after 19 research flights. A shorter follow-on program involving seven flights began in September 1998 and concluded in April 1999. All flights were conducted in Russia from Tupolev's facility at the Zhukovsky Air Development Center near Moscow. The centerpiece of the research program was the Tu 144LL, a first-generation Russian supersonic jetliner that was modified by its developer/builder, Tupolev ANTK (aviatsionnyy nauchno-tekhnicheskiy kompleks-roughly, aviation technical complex), into a flying laboratory for supersonic research. Using the Tu-144LL to conduct flight research experiments, researchers compared full-scale supersonic aircraft flight data with results from models in wind tunnels, computer-aided techniques, and other flight tests. The experiments provided unique aerodynamic, structures, acoustics, and operating environment data on supersonic passenger aircraft. Data collected from the research program was being used to develop the technology base for a proposed future American-built supersonic jetliner. Although actual development of such an advanced supersonic transport (SST) is currently on hold, commercial aviation experts estimate that a market for up to 500 such aircraft could develop by the third decade of the 21st Century. The Tu-144LL used in the NASA-sponsored research program was a 'D' model with different engines than were used in production-model aircraft. Fifty experiments were proposed

  13. Russian Tu-144LL SST Joint NASA Flying Laboratory - Flight November 29, 1996

    NASA Technical Reports Server (NTRS)

    1996-01-01

    The modified Tupolev Tu-144LL supersonic flying laboratory during a test flight from the Zhukovsky Airfield near Moscow, Russia. The 'LL' stands for Letayuschaya Laboratoriya, which means Flying Laboratory. NASA teamed with American and Russian aerospace industries for an extended period in a joint international research program featuring the Russian-built Tu-144LL supersonic aircraft. The object of the program was to develop technologies for a proposed future second-generation supersonic airliner to be developed in the 21st Century. The aircraft's initial flight phase began in June 1996 and concluded in February 1998 after 19 research flights. A shorter follow-on program involving seven flights began in September 1998 and concluded in April 1999. All flights were conducted in Russia from Tupolev's facility at the Zhukovsky Air Development Center near Moscow. The centerpiece of the research program was the Tu 144LL, a first-generation Russian supersonic jetliner that was modified by its developer/builder, Tupolev ANTK (aviatsionnyy nauchno-tekhnicheskiy kompleks-roughly, aviation technical complex), into a flying laboratory for supersonic research. Using the Tu-144LL to conduct flight research experiments, researchers compared full-scale supersonic aircraft flight data with results from models in wind tunnels, computer-aided techniques, and other flight tests. The experiments provided unique aerodynamic, structures, acoustics, and operating environment data on supersonic passenger aircraft. Data collected from the research program was being used to develop the technology base for a proposed future American-built supersonic jetliner. Although actual development of such an advanced supersonic transport (SST) is currently on hold, commercial aviation experts estimate that a market for up to 500 such aircraft could develop by the third decade of the 21st Century. The Tu-144LL used in the NASA-sponsored research program was a 'D' model with different engines than were

  14. A Qualitative Piloted Evaluation of the Tupolev Tu-144 Supersonic Transport

    NASA Technical Reports Server (NTRS)

    Rivers, Robert A.; Jackson, E. Bruce; Fullerton, C. Gordon; Cox, Timothy H.; Princen, Norman H.

    2000-01-01

    Two U.S. research pilots evaluated the Tupolev Tu-144 supersonic transport aircraft on three dedicated flights: one subsonic and two supersonic profiles. The flight profiles and maneuvers were developed jointly by Tupolev and U.S. engineers. The vehicle was found to have unique operational and flight characteristics that serve as lessons for designers of future supersonic transport aircraft. Vehicle subsystems and observed characteristics are described as are flight test planning and ground monitoring facilities. Maneuver descriptions and extended pilot narratives for each flight are included as appendices.

  15. Youyou Tu: significance of winning the 2015 Nobel Prize in Physiology or Medicine

    PubMed Central

    Liu, Wenxiu

    2016-01-01

    Youyou Tu, a female scientist at the China Academy of Traditional Chinese Medicine in Beijing, is the first Chinese winner of the Nobel Prize in Physiology or Medicine. Based on the study of recipes which had been used for thousands of years to treat fever, Tu’s group discovered that the plant artemesia annua, sweet wormwood, showed substantial inhibition of rodent malaria parasites. Her achievement and experience have inspired other researchers and emphasized the development of traditional Chinese medicine. Her award has led to a heated discussion about scientific research investment, fair treatment of research staff, and intellectual property right (IPR) protection in China. PMID:26885485

  16. Protein synthesis alongation factors EF-Tu and eEF1A: biosynthesis, functions and application in the improvement of heat tolerance in plants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Protein synthesis elongation factors EF-Tu and eEF1A (EFs) represent a group of highly conserved and abundant GTPases with an important role in transporting the aminoacyl-tRNA complex to the A site of the ribosome during elongation phase of translation. EF-Tu proteins are located in bacteria and, du...

  17. Immunoregulation in experimental disseminated histoplasmosis: flow microfluorometry (FMF) studies of the Thy and Lyt phenotypes of T lymphocytes from infected mice.

    PubMed

    Watson, S R; Miller, T B; Redington, T J; Bullock, W E

    1983-08-01

    Previous studies have shown that mice infected i.v. with 6 X 10(5) yeast phase Histoplasma capsulatum (Hc) develop suppressed immune responses during weeks 1 to 4 of infection but that by weeks 8 to 12 of infection these responses return to normal. In this study total and differential cell counts showed that as early as the third day of infection there was a marked reduction in the number of lymphocytes recovered from the peripheral blood, bone marrow, and thymus of infected animals. Concomitantly, there was an increase in the number of splenic lymphocytes. By day 28 both the total and differential cell counts were similar in both infected and normal animals. Flow microfluorometric (FMF) studies comparing the Thy-1.2, Lyt-1, Lyt-2, and surface immunoglobulin (slg) phenotypes of lymphocytes from normal and infected mice were performed. Between days 5 and 7 the thymocytes from infected mice displayed a higher relative fluorescence intensity (RFI) of the Thy-1.2 marker than normal thymocytes, whereas at day 10, the RFI was less than that of normal thymic lymphocytes. Between days 7 and 10 of infection the RFI of the Lyt-2 marker was less on thymocytes from Hc-infected mice; however, there was no change in the Lyt-1 marker. Examination of these lymphocyte markers in blood, spleen, and mesenteric lymph nodes showed that there were decreases in the RFI of both the Thy-1.2 and Lyt-2 between days 5 and 10 of infection. No changes were observed in the Lyt-1 or slg markers. By day 28 there were no differences between the normal and infected mice with respect to any surface marker in any of the organs studied. In other experiments, the effect of adrenalectomy before infection on these surface markers was studied. Absolute numbers of Thy-1.2+, Lyt-1+, and Lyt-2+ cells were significantly increased in the spleen and significantly decreased in the thymus and peripheral blood of infected mice relative to normal controls. These studies suggest that there is a migration of cells

  18. Divergence among Genes Encoding the Elongation Factor Tu of Yersinia Species▿

    PubMed Central

    Isabel, Sandra; Leblanc, Éric; Boissinot, Maurice; Boudreau, Dominique K.; Grondin, Myrian; Picard, François J.; Martel, Eric A.; Parham, Nicholas J.; Chain, Patrick S. G.; Bader, Douglas E.; Mulvey, Michael R.; Bryden, Louis; Roy, Paul H.; Ouellette, Marc; Bergeron, Michel G.

    2008-01-01

    Elongation factor Tu (EF-Tu), encoded by tuf genes, carries aminoacyl-tRNA to the ribosome during protein synthesis. Duplicated tuf genes (tufA and tufB), which are commonly found in enterobacterial species, usually coevolve via gene conversion and are very similar to one another. However, sequence analysis of tuf genes in our laboratory has revealed highly divergent copies in 72 strains spanning the genus Yersinia (representing 12 Yersinia species). The levels of intragenomic divergence between tufA and tufB sequences ranged from 8.3 to 16.2% for the genus Yersinia, which is significantly greater than the 0.0 to 3.6% divergence observed for other enterobacterial genera. We further explored tuf gene evolution in Yersinia and other Enterobacteriaceae by performing directed sequencing and phylogenetic analyses. Phylogenetic trees constructed using concatenated tufA and tufB sequences revealed a monophyletic genus Yersinia in the family Enterobacteriaceae. Moreover, Yersinia strains form clades within the genus that mostly correlate with their phenotypic and genetic classifications. These genetic analyses revealed an unusual divergence between Yersinia tufA and tufB sequences, a feature unique among sequenced Enterobacteriaceae and indicative of a genus-wide loss of gene conversion. Furthermore, they provided valuable phylogenetic information for possible reclassification and identification of Yersinia species. PMID:18790860

  19. Low Order Equivalent System Identification for the Tu-144LL Supersonic Transport Aircraft

    NASA Technical Reports Server (NTRS)

    Morelli, Eugene A.

    2000-01-01

    Low order equivalent system models were identified from flight test data for the Tu-144LL supersonic transport aircraft. Flight test maneuvers were executed by Russian and American test Pilots flying the aircraft from Zhukovsky airfield outside Moscow, Russia. Flight tests included longitudinal and lateral / directional maneuvers at supersonic cruise flight conditions. Piloted frequency sweeps and multi-step maneuvers were used to generate data for closed loop low order equivalent system modeling. Model parameters were estimated using a flexible. high accuracy Fourier transform and an equation error / output error (EE/OE) formulation in the frequency domain. Results were compared to parameter estimates obtained using spectral estimation and subsequent least squares fit to frequency response data in Bode plots. Modeling results from the two methods agreed well for both a frequency sweep and multiple concatenated multi-step maneuvers. For a single multi-step maneuvers the EE/OE method gave a better model fit with improved prediction capability. A summary of closed loop low order equivalent system identification results for the Tu-144LL, including estimated parameters, standard errors, and flying qualities level predictions, were computed and tabulated.

  20. TU Comae Berenices: Blazhko RR Lyrae Star in a Potential Binary System

    NASA Astrophysics Data System (ADS)

    de Ponthière, P.; Hambsch, F.-J.; Menzies, K.; Sabo, R.

    2016-06-01

    We present the results of a photometry campaign of TU Com performed over a five-year time span. The analysis showed that the possible Blazhko period of 75 days published by the General Catalogue of Variable Stars is not correct. We identified two Blazhko periods of 43.6 and 45.5 days. This finding is based on measurement of 124 light maxima. A spectral analysis of the complete light curve confirmed these two periods. Besides the Blazhko amplitude and phase modulations, another long term periodic phase variation has been identified. This long term periodic variation affects the times of maximum light only and can be attributed to a light-travel time effect due to orbital motion of a binary system. The orbital parameters have been estimated by a nonlinear least-square fit applied to the set of (O-C) values. The Levenberg-Marquart algorithm has been used to perform the nonlinear least-square fit. The tentative orbital parameters include an orbital period of 1676 days, a minimal semi-major axis of 1.55 AU and a small eccentricity of 0.22. The orbital parameter estimation also used 33 (O-C) values obtained from the SWASP survey database. Spectroscopic radial velocity measurements are needed to confirm this binarity. If confirmed, TU Com would be the first Blazhko RR Lyrae star detected in a binary system.

  1. Flight Test Measurements From The Tu-144LL Structure/Cabin Noise Follow-On Experiment

    NASA Technical Reports Server (NTRS)

    Rizzi, Stephen A.; Rackl, Robert G.; Andrianov, Eduard V.

    2000-01-01

    This follow-on flight experiment on the TU-144LL Supersonic Flying Laboratory, conducted during the period September 1998 to April 1999, was a continuation of previous Structure/Cabin Noise Experiment 2.1. Data was obtained over a wide range of altitudes and Mach numbers. Measured were: turbulent boundary layer pressure fluctuations on the fuselage over its length; structural response on skin panels using accelerometers; and flow direction over three windows using 'flow cones'. The effect of steps in the flow was also measured using two window blank pairs; each pair bridged by a plate which created small sharp forward and aft facing steps. The effect of transducer flushness with the exterior surface was also measured during flight. Height test points were chosen to cover much of the TU-144's flight envelope, as well as to obtain as large a unit Reynolds number range as possible at various Mach numbers: takeoff, subsonic, transonic, and supersonic cruise conditions up to Mach 2. Data on engine runups and background noise were acquired on the ground. The data in the form of time histories of the acoustic signals, together with auxiliary data and basic MATLAB processing modules, are available on CD-R disks.

  2. Modeling study of carbonate decomposition in LLNL`s 4TU pilot oil shale retort

    SciTech Connect

    Thorsness, C.B.

    1994-10-14

    Lawrence Livermore National Laboratory`s (LLNL) 4 tonne-per-day oil shale Pilot Retort (4TU-Pilot) has been modeled to study the degree of carbonate decomposition occurring in the process. The modeling uses a simplified version of the processes occurring in the retort to allow parametric studies to be performed. The primary focus of the work is on the sensitivity of computed carbonate decomposition to the assumed manner in which solid material leaves the retort. It was found that for a variety of assumptions about solid passage and evolution within the process the computed carbonate decomposition varied by only a few percent. It was also determined that using available kinetic expressions based on literature data led to a consistent underestimate of the carbonate decomposition, from 12--17% low on an absolute basis and on a relative basis as much as a factor of seven times too low. A simplified kinetic expression based on limited data from laboratory experiments on the same shale as used in the 4TU-Pilot run was also employed and found to match the pilot results fairly well.

  3. Estimation of Handling Qualities Parameters of the Tu-144 Supersonic Transport Aircraft from Flight Test Data

    NASA Technical Reports Server (NTRS)

    Curry, Timothy J.; Batterson, James G. (Technical Monitor)

    2000-01-01

    Low order equivalent system (LOES) models for the Tu-144 supersonic transport aircraft were identified from flight test data. The mathematical models were given in terms of transfer functions with a time delay by the military standard MIL-STD-1797A, "Flying Qualities of Piloted Aircraft," and the handling qualities were predicted from the estimated transfer function coefficients. The coefficients and the time delay in the transfer functions were estimated using a nonlinear equation error formulation in the frequency domain. Flight test data from pitch, roll, and yaw frequency sweeps at various flight conditions were used for parameter estimation. Flight test results are presented in terms of the estimated parameter values, their standard errors, and output fits in the time domain. Data from doublet maneuvers at the same flight conditions were used to assess the predictive capabilities of the identified models. The identified transfer function models fit the measured data well and demonstrated good prediction capabilities. The Tu-144 was predicted to be between level 2 and 3 for all longitudinal maneuvers and level I for all lateral maneuvers. High estimates of the equivalent time delay in the transfer function model caused the poor longitudinal rating.

  4. Structure of ThiM from Vitamin B1 biosynthetic pathway of Staphylococcus aureus – Insights into a novel pro-drug approach addressing MRSA infections

    PubMed Central

    Drebes, Julia; Künz, Madeleine; Windshügel, Björn; Kikhney, Alexey G.; Müller, Ingrid B.; Eberle, Raphael J.; Oberthür, Dominik; Cang, Huaixing; Svergun, Dmitri I.; Perbandt, Markus; Betzel, Christian; Wrenger, Carsten

    2016-01-01

    Infections caused by the methicillin-resistant Staphylococcus aureus (MRSA) are today known to be a substantial threat for global health. Emerging multi-drug resistant bacteria have created a substantial need to identify and discover new drug targets and to develop novel strategies to treat bacterial infections. A promising and so far untapped antibiotic target is the biosynthesis of vitamin B1 (thiamin). Thiamin in its activated form, thiamin pyrophosphate, is an essential co-factor for all organisms. Therefore, thiamin analogous compounds, when introduced into the vitamin B1 biosynthetic pathway and further converted into non-functional co-factors by the bacterium can function as pro-drugs which thus block various co-factor dependent pathways. We characterized one of the key enzymes within the S. aureus vitamin B1 biosynthetic pathway, 5-(hydroxyethyl)-4-methylthiazole kinase (SaThiM; EC 2.7.1.50), a potential target for pro-drug compounds and analyzed the native structure of SaThiM and complexes with the natural substrate 5-(hydroxyethyl)-4-methylthiazole (THZ) and two selected substrate analogues. PMID:26960569

  5. Complete nucleotide sequence of the Escherichia coli recC gene and of the thyA-recC intergenic region.

    PubMed Central

    Finch, P W; Wilson, R E; Brown, K; Hickson, I D; Tomkinson, A E; Emmerson, P T

    1986-01-01

    The nucleotide sequence of a 6,000 bp region of the E. coli chromosome that includes the 3' end of the coding region for the thyA gene and the entire recC gene has been determined. The proposed coding region for the RecC protein is 3369 nucleotides long, which would encode a polypeptide consisting of 1122 amino acids with a calculated molecular mass of 129 kDa. Mung bean nuclease mapping of a recC specific transcript produced in vivo indicates that transcription of recC is initiated 80 bp upstream of the translational start point. A weak promoter sequence situated 5' to the transcription initiation point has been identified. In the 1953 bp thyA-recC intergenic region there are three open reading frames that would code for polypeptides of molecular mass 30 kDa, 13.5 kDa and 12 kDa, respectively. Although the first and third of these open reading frames are preceded by possible ribosome binding sites, no obvious promoter sequences could be identified. Moreover, transcripts for these reading frames could not be detected. Images PMID:3520484

  6. In vivo nonlinear imaging of corneal structures with special focus on BALB/c and streptozotocin-diabetic Thy1-YFP mice.

    PubMed

    Ehmke, Tobias; Leckelt, Janine; Reichard, Maria; Weiss, Heike; Hovakimyan, Marina; Heisterkamp, Alexander; Stachs, Oliver; Baltrusch, Simone

    2016-05-01

    Two-photon microscopy (TPM) allows high contrast imaging at a subcellular resolution scale. In this work, the microscopy technique was applied to visualize corneal structures in two mouse models (BALB/c and B6.Cg-Tg(Thy1-YFP)16Jrs/J) in vivo. In particular, the transgenic Thy1-YFP mice expressing the yellow fluorescent protein (YFP) in all motor and sensory neurons had been used for investigating the nerve fiber density in healthy and streptozotocin-diabetic mice. This model is clinically relevant since patients suffering from diabetes mellitus have a high risk to develop small fiber neuropathy. Nonlinear laser scanning microscopy displayed a reduction of nerve fiber density in streptozotocin-diabetic versus healthy mice and confirmed data obtained by confocal laser scanning microscopy (CLSM). In recent years, corneal CLSM was proved to be an appropriate non-invasive tool for an early diagnosis of diabetic neuropathy. Nevertheless, validation of the CLSM method for the clinical routine is currently a matter of investigation and requires confirmation by further studies and complementary techniques. Thus, the present study provides further evidence of corneal confocal microscopy as a promising technique for non-invasive detection of diabetic neuropathy. Information derived from these experiments may become clinically relevant and help to develop new drugs for treatment of diabetic neuropathy. PMID:26677758

  7. Transgene therapy for rat anti-Thy1.1 glomerulonephritis via mesangial cell vector with a polyethylenimine/decorin nanocomplex

    NASA Astrophysics Data System (ADS)

    Sun, Jian-Yong; Sun, Yu; Wu, Hui-Juan; Zhang, Hong-Xia; Zhao, Zhong-Hua; Chen, Qi; Zhang, Zhi-Gang

    2012-08-01

    Polyethylenimine (PEI), a cationic polymer, is one of the most efficient non-viral vectors for transgene therapy. Decorin (DCN), a leucine-rich proteoglycan secreted by glomerular mesangial cells (MC), is a promising anti-fibrotic agent for the treatment of glomerulonephritis. In this study, we used PEI-DCN nanocomplexes with different N/P ratios to transfect MC in vitro and deliver the MC vector with PEI-DCN expressing into rat anti-Thy1.1 nephritis kidney tissue via injection into the left renal artery in vivo. The PEI-plasmid DNA complex at N/P 20 had the highest level of transfection efficiency and the lowest level of cytotoxicity in cultured MC. Following injection, the ex vivo gene was transferred successfully into the glomeruli of the rat anti-Thy1.1 nephritis model by the MC vector with the PEI-DCN complex. The exogenous MC with DCN expression was located mainly in the mesangium and the glomerular capillary. Over-expression of DCN in diseased glomeruli could result in the inhibition of collagen IV deposition and MC proliferation. The pathological changes of rat nephritis were alleviated following injection of the vector. These findings demonstrate that the DCN gene delivered by the PEI-DNA nanocomplex with the MC vector is a promising therapeutic method for the treatment of glomerulonephritis.

  8. The effects of PTK787/ZK222584, an inhibitor of VEGFR and PDGFRβ pathways, on intussusceptive angiogenesis and glomerular recovery from Thy1.1 nephritis.

    PubMed

    Wnuk, Monika; Hlushchuk, Ruslan; Tuffin, Gérald; Huynh-Do, Uyen; Djonov, Valentin

    2011-04-01

    The aim of our study was to investigate the phenomenon of intussusceptive angiogenesis with a focus on its molecular regulation by vascular endothelial growth factor receptor (VEGFR)/platelet-derived growth factor receptor β (PDGFRβ) pathways and biological significance for glomerular recovery after acute injury. Glomerular healing by intussusception was examined in a particular setting of Thy1.1 nephritis, where the lysis of mesangial cells results in an initial collapse and successive rebuilding of glomerular capillary structure. Restoration of capillary structure after induction of Thy1.1 nephritis occurred by intussusceptive angiogenesis resulting in i) rapid expansion of the capillary plexus with reinstatement of the glomerular filtration surface and ii) restoration of the archetypical glomerular vascular pattern. Glomerular capillaries of nephritic rats after combined VEGFR2 and PDGFRβ inhibition by PTK787/ZK222584 (PTK/ZK) were tortuous and irregular. However, the onset of intussusceptive angiogenesis was influenced only after long-term PTK/ZK treatment, providing an important insight into differential molecular regulation between sprouting and intussusceptive angiogenesis. PTK/ZK treatment abolished α-smooth muscle actin and tensin expression by injured mesangial cells, impaired glomerular filtration of microspheres, and led to the reduction of glomerular volume and the presence of multiple hemorrhages detectable in the tubular system. Collectively, treatment of nephritic patients with PTK/ZK compound is not recommended. PMID:21435466

  9. A novel high-throughput and quantitative method based on visible color shifts for screening Bacillus subtilis THY-15 for surfactin production.

    PubMed

    Yang, Huan; Yu, Huimin; Shen, Zhongyao

    2015-08-01

    A novel chromatic visible screening method using bromothymol blue (BTB) as a color indicator and cetylpyridinium chloride (CPC) as a mediator was constructed to obtain the high titer surfactin-producing strains. The reliability and quantification accuracy of color shift were also confirmed. Regular chromatic responses from faint yellow-green to dark green and bright blue reflected the different ranges of surfactin concentrations. Moreover, the quantitative accuracy of surfactin quantification in the range of 100-500 mg/L was verified by reverse-phase high-performance liquid chromatography (RP-HPLC) using different fermentation supernatant samples. Using this CPC-BTB method, a superior surfactin producer, Bacillus subtilis THY-15, was successfully screened. The producer's surfactin (Srf) titer reached 1240 mg/L. RP-HPLC analysis of THY-15 revealed four surfactin isoforms. As identified by amino acid analysis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis, the isoforms of surfactin in fraction 1, 2 and 4 had the same circular peptide sequence of Glu-Leu-Leu-Val-Asp-Leu-Leu but different iso-C13, C14 and C15 fatty acid chains, but the isoform in fraction 3 possessed a special peptide sequence of Glu-Val-Leu-Leu-Asp-Leu-Val. PMID:26065390

  10. Structure of ThiM from Vitamin B1 biosynthetic pathway of Staphylococcus aureus – Insights into a novel pro-drug approach addressing MRSA infections

    NASA Astrophysics Data System (ADS)

    Drebes, Julia; Künz, Madeleine; Windshügel, Björn; Kikhney, Alexey G.; Müller, Ingrid B.; Eberle, Raphael J.; Oberthür, Dominik; Cang, Huaixing; Svergun, Dmitri I.; Perbandt, Markus; Betzel, Christian; Wrenger, Carsten

    2016-03-01

    Infections caused by the methicillin-resistant Staphylococcus aureus (MRSA) are today known to be a substantial threat for global health. Emerging multi-drug resistant bacteria have created a substantial need to identify and discover new drug targets and to develop novel strategies to treat bacterial infections. A promising and so far untapped antibiotic target is the biosynthesis of vitamin B1 (thiamin). Thiamin in its activated form, thiamin pyrophosphate, is an essential co-factor for all organisms. Therefore, thiamin analogous compounds, when introduced into the vitamin B1 biosynthetic pathway and further converted into non-functional co-factors by the bacterium can function as pro-drugs which thus block various co-factor dependent pathways. We characterized one of the key enzymes within the S. aureus vitamin B1 biosynthetic pathway, 5-(hydroxyethyl)-4-methylthiazole kinase (SaThiM; EC 2.7.1.50), a potential target for pro-drug compounds and analyzed the native structure of SaThiM and complexes with the natural substrate 5-(hydroxyethyl)-4-methylthiazole (THZ) and two selected substrate analogues.

  11. FdhTU-modulated formate dehydrogenase expression and electron donor availability enhance recovery of Campylobacter jejuni following host cell infection

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Analysis of Campylobacter jejuni fdhTU reveals a role in formate dehydrogenase activity and implications for electron donor requirements during the pathogen-host cell interaction. Campylobacter jejuni is a foodborne bacterial pathogen which colonizes the intestinal tract and causes severe gastroent...

  12. Trim25 Is an RNA-Specific Activator of Lin28a/TuT4-Mediated Uridylation

    PubMed Central

    Choudhury, Nila Roy; Nowak, Jakub S.; Zuo, Juan; Rappsilber, Juri; Spoel, Steven H.; Michlewski, Gracjan

    2014-01-01

    Summary RNA binding proteins have thousands of cellular RNA targets and often exhibit opposite or passive molecular functions. Lin28a is a conserved RNA binding protein involved in pluripotency and tumorigenesis that was previously shown to trigger TuT4-mediated pre-let-7 uridylation, inhibiting its processing and targeting it for degradation. Surprisingly, despite binding to other pre-microRNAs (pre-miRNAs), only pre-let-7 is efficiently uridylated by TuT4. Thus, we hypothesized the existence of substrate-specific cofactors that stimulate Lin28a-mediated pre-let-7 uridylation or restrict its functionality on non-let-7 pre-miRNAs. Through RNA pull-downs coupled with quantitative mass spectrometry, we identified the E3 ligase Trim25 as an RNA-specific cofactor for Lin28a/TuT4-mediated uridylation. We show that Trim25 binds to the conserved terminal loop (CTL) of pre-let-7 and activates TuT4, allowing for more efficient Lin28a-mediated uridylation. These findings reveal that protein-modifying enzymes, only recently shown to bind RNA, can guide the function of canonical ribonucleoprotein (RNP) complexes in cis, thereby providing an additional level of specificity. PMID:25457611

  13. A guidelines handbook for GPR surveys in tunnels: a COST Action TU1208 contribution

    NASA Astrophysics Data System (ADS)

    Bianchini Ciampoli, Luca; Alani, Amir M.; Pajewski, Lara; Benedetto, Andrea; Loizos, Andreas; Tosti, Fabio

    2016-04-01

    A significant open issue concerning the reliability of geophysical methods and in particular of ground penetrating radar (GPR), both in research and professional context, is a general lack of international standards. This is a major problem to be faced, in order to gain scientific strictness for the GPR practices, and to easily extend to the international community the results achieved within the area of single virtuous countries. Producing international guidelines can represent an important step forward, in this sense. In the memorandum of understanding of the COST Action TU1208 is clearly stated that one of the main purposes of the Action is the "development of innovative protocols and guidelines which will be published in a handbook and constitute a basis for European Standards, for an effective GPR application in CE tasks; safety, economic and financial criteria will be integrated within the protocols". Of course this is not a simple task to be accomplished. Firstly, survey procedures are highly dependent on the objective of the survey itself. On the basis of the objective of each geophysical test, the GPR system, the antenna configuration, and even the processing procedures may change. Besides, these procedures are also influenced by the environmental conditions in which the tests are performed. This affects several aspects spanning from hardware to software, but including, for instance, also safety issues. Due to these reasons, one of the main goal of the COST Action TU1208 is the development of several guidelines related to the main applications of GPR in the field of civil engineering. In this work, the structure of a guidelines handbook for GPR activities in tunnels is outlined. In the first sections, the principal references in the field are provided, and the most common GPR equipment and complementary technologies are described. Subsequently, the survey methodologies are explained. Particular attention is paid to the preliminary activities to be carried

  14. Preventive Effect of TU-100 on a Type-2 Model of Colitis in Mice: Possible Involvement of Enhancing Adrenomedullin in Intestinal Epithelial Cells

    PubMed Central

    Kono, Toru; Miura, Naoko

    2013-01-01

    Purpose. Crohn's disease (CD) and ulcerative colitis (UC), the two major forms of inflammatory bowel disease (IBD), have histopathologically and immunologically different characteristics. We previously reported that a traditional Japanese medicine, daikenchuto (TU-100), ameliorated a trinitrobenzenesulfonic acid- (TNBS-) induced type-1 model colitis exhibiting histopathological features of CD through adrenomedullin (ADM) enhancement. Our current aims were to examine whether TU-100 ameliorates a type-2 model colitis that histologically resembles UC and identify the active ingredients. Methods. TU-100 was administered orally to mice with oxazolone- (OXN-) induced type-2 model colitis. The morbidity was evaluated by body weight loss and the macroscopic score of colonic lesions. ADM was quantified using an EIA kit. Results. TU-100 prevented weight loss and colon ulceration. ADM production by intestinal epithelial cells was increased by TU-100 addition. Screening to identify active ingredients showed that [6]-shogaol and hydroxy α-sanshool enhanced ADM production. Conclusions. TU-100 exerted a protective effect in OXN-induced type-2 model colitis, indicating that TU-100 may be a beneficial agent for treatment of UC. PMID:24348533

  15. Ecology of Phlebotomine Sand Flies in the Rural Community of Mont Rolland (Thiès Region, Senegal): Area of Transmission of Canine Leishmaniasis

    PubMed Central

    Senghor, Massila W.; Faye, Malick N.; Faye, Babacar; Diarra, Karamoko; Elguero, Eric; Gaye, Oumar

    2011-01-01

    Background Different epidemiological studies previously indicated that canine leishmaniasis is present in the region of Thiès (Senegal). However, the risks to human health, the transmission cycle and particularly the implicated vectors are unknown. Methodology/Principal Findings To improve our knowledge on the population of phlebotomine sand flies and the potential vectors of canine leishmaniasis, sand flies were collected using sticky traps, light traps and indoor spraying method using pyrethroid insecticides in 16 villages of the rural community of Mont Rolland (Thiès region) between March and July 2005. The 3788 phlebotomine sand flies we collected (2044 males, 1744 females) were distributed among 9 species of which 2 belonged to the genus Phlebotomus: P. duboscqi (vector of cutaneous leishmaniasis in Senegal) and P. rodhaini. The other species belonged to the genus Sergentomyia: S. adleri, S. clydei, S. antennata, S. buxtoni, S. dubia, S. schwetzi and S. magna. The number of individuals and the species composition differed according to the type of trap, suggesting variable, species-related degrees of endophily or exophily. The two species of the genus Phlebotomus were markedly under-represented in comparison to the species of the genus Sergentomyia. This study also shows a heterogeneous spatial distribution within the rural community that could be explained by the different ecosystems and particularly the soil characteristics of this community. Finally, the presence of the S. dubia species appeared to be significantly associated with canine leishmaniasis seroprevalence in dogs. Conclusions/Significance Our data allow us to hypothesize that the species of the genus Sergentomyia and particularly the species S. dubia and S. schwetzi might be capable of transmitting canine leishmaniasis. These results challenge the dogma that leishmaniasis is exclusively transmitted by species of the genus Phlebotomus in the Old World. This hypothesis should be more thoroughly

  16. Translational activities of EF-Tu [G222D] which cannot be reconciled with the classical scheme of the polypeptide chain elongation cycle.

    PubMed

    Talens, A; Boon, K; Kraal, B; Bosch, L

    1996-08-23

    We have developed a cell-free system of E. coli that enables us to study the in vitro translation of natural mRNA mediated by wild-type or mutant EF-Tu. Various mutant EF-Tu species have been analyzed, one of which, EF-Tu [G222D], appeared to be virtually unable to mediate the translation of natural mRNA. Since this mutant factor is able to participate in translation in vivo by suppressing nonsense and frameshift mutations in cooperation with EF-Tu [A375T], a revision of the generally accepted scheme of the elongation cycle has been proposed (Bosch, L., Vijgenboom, E., & Zeef, L.A.H., 1996, Biochemistry 36). PMID:8780718

  17. Vacuolar Ca(2+) uptake.

    PubMed

    Pittman, Jon K

    2011-08-01

    Calcium transporters that mediate the removal of Ca(2+) from the cytosol and into internal stores provide a critical role in regulating Ca(2+) signals following stimulus induction and in preventing calcium toxicity. The vacuole is a major calcium store in many organisms, particularly plants and fungi. Two main pathways facilitate the accumulation of Ca(2+) into vacuoles, Ca(2+)-ATPases and Ca(2+)/H(+) exchangers. Here I review the biochemical and regulatory features of these transporters that have been characterised in yeast and plants. These Ca(2+) transport mechanisms are compared with those being identified from other vacuolated organisms including algae and protozoa. Studies suggest that Ca(2+) uptake into vacuoles and other related acidic Ca(2+) stores occurs by conserved mechanisms which developed early in evolution. PMID:21310481

  18. Development of SAP-DoA techniques for GPR data processing within COST Action TU1208

    NASA Astrophysics Data System (ADS)

    Meschino, Simone; Pajewski, Lara; Marciniak, Marian

    2016-04-01

    This work focuses on the use of Sub-Array Processing (SAP) and Direction of Arrival (DoA) approaches for the processing of Ground-Penetrating Radar data, with the purpose of locating metal scatterers embedded in concrete or buried in the ground. Research activities have been carried out during two Short-Term Scientific Missions (STSMs) funded by the COST (European COoperation in Science and Technology) Action TU1208 "Civil Engineering Applications of Ground Penetrating Radar" in May 2015 and January 2016. In applications involving smart antennas and in the presence of several transmitters operating simultaneously, it is important for a receiving array to be able to estimate the Direction of Arrival (DoA) of the incoming signals, in order to decipher how many emitters are present and predict their positions. A number of methods have been devised for DoA estimation: the MUltiple SIgnal Classification (MUSIC) and Estimation of Signal Parameters via Rotational Invariance Technique (ESPRIT) are amongst the most popular ones [1]. In the scenario considered by us, the electromagnetic sources are the currents induced on metal elements embedded in concrete or buried in the ground. GPR radargrams are processed, to estimate the DoAs of the electric field back-scattered by the sought targets. In order to work in near-field conditions, a sub-array processing (SAP) approach is adopted: the radargram is partitioned in sub-radargrams composed of few A-scans each, the dominant DoA is predicted for each sub-radargram. The estimated angles are triangulated, obtaining a set of crossings with intersections condensed around object locations. This pattern is filtered, in order to remove a noisy background of unwanted crossings, and is processed by applying the statistical procedure described in [2]. We tested our approach on synthetic GPR radargrams, obtained by using the freeware simulator gprMax implementing the Finite-Difference Time-Domain method [3]. In particular, we worked with

  19. A relational-tubular (ReTu) deformable model for vasculature quantification of zebrafish embryo from microangiography image series.

    PubMed

    Feng, Jun; Ip, Horace H S; Cheng, Shuk H; Chan, Po K

    2004-09-01

    Embryonic cardiovascular system plays a vital role in embryonic development of human and animal. In this work, we introduce a novel deformable model, which we called Relational-tubular (ReTu) deformable model for segmenting and quantifying the embryonic vasculature of zebrafish embryo from microangiography image series. Particularly, to incorporate additional constraints on the spatial relationships among vessel branches, we introduce a new energy term called relation energy into the model energy function. This energy item acts as a repulsion force between neighboring vessels during the deformation to encourage them to move towards their respective volume data. Using the ReTu deformable model, the deformation process is an iterative two-stage procedure: vascular axis deformation and vascular surface deformation. The efficiency and robustness of this approach are demonstrated by experiments which show that satisfactory quantifications of the vasculature can be obtained after 3-4 iterations. PMID:15294311

  20. Formulations of the endophytic bacterium Bacillus subtilis Tu-100 suppress Sclerotinia sclerotiorum on oilseed rape and improve plant vigor in field trials conducted at separate locations.

    PubMed

    Hu, Xiaojia; Roberts, Daniel P; Maul, Jude E; Emche, Sarah E; Liao, Xing; Guo, Xuelan; Liu, Yeying; McKenna, Laurie F; Buyer, Jeffrey S; Liu, Shengyi

    2011-07-01

    Sclerotinia sclerotiorum causes serious yield losses in crops in the People's Republic of China. Two formulations of oilseed rape seed containing the bacterium Bacillus subtilis Tu-100 were evaluated for suppression of this pathogen in field trials conducted at two independent locations. The pellet formulation significantly reduced disease (incidence and disease index) and increased plant dry mass, while the wrap formulation significantly reduced disease incidence and significantly increased plant dry mass at both field locations. Mean seed yield per 120 plants with both formulations of isolate Tu-100 was significantly greater than the appropriate controls, but at only one of the locations. Both formulations provided stable B. subtilis Tu-100 biomass (≥10(5) CFU·g(-1)) and seed germination (≥85%) over a 6 month period at room temperature. Polymerase chain reaction and DNA sequence analysis identified ituC and ituD, and bacAB and bacD in the genome of isolate Tu-100. These genes are involved in the biosynthesis of iturin and bacilysin. Iturin was detected in culture filtrates from isolate Tu-100, with thin layer chromatography. Detection of bacilysin was not attempted. Experiments reported here indicate the commercial viability of B. subtilis Tu-100 for suppression of S. sclerotiorum on oilseed rape. PMID:21767217

  1. Interaction of helix D of elongation factor Tu with helices 4 and 5 of protein L7/12 on the ribosome.

    PubMed

    Kothe, Ute; Wieden, Hans-Joachim; Mohr, Dagmar; Rodnina, Marina V

    2004-03-01

    Elongation factor Tu (EF-Tu) promotes binding of aminoacyl-tRNA to the A site of the ribosome. Here, we report the effects of mutations in helix D of EF-Tu and in the C-terminal domain of L7/12 on the kinetics of A-site binding. Reaction rates were measured by stopped-flow and quench-flow techniques. The rates of A-site binding were decreased by mutations at positions 144, 145, 148, and 152 in helix D of EF-Tu as well as at positions 65, 66, 69, 70, 73, and 84 in helices 4 and 5 of L7/12. The effect was due primarily to the lower association rate constant of ternary complex binding to the ribosome. These results suggest that helix D of EF-Tu is involved in an initial transient contact with helices 4 and 5 of L7/12 that promotes ternary complex binding to the ribosome. By analogy to the interaction of helix D of EF-Tu with the N-terminal domain of EF-Ts, the contact area is likely to consist of a hydrophobic patch flanked by two salt-bridges. PMID:15037065

  2. BnSGS3 Has Differential Effects on the Accumulation of CMV, ORMV and TuMV in Oilseed Rape.

    PubMed

    Chen, Quan; Wang, Jie; Hou, Mingsheng; Liu, Shengyi; Huang, Junyan; Cai, Li

    2015-08-01

    Virus diseases greatly affect oilseed rape (Brassica napus) production. Investigating antiviral genes may lead to the development of disease-resistant varieties of oilseed rape. In this study, we examined the effects of the suppressor of gene silencing 3 in Brassica napus (BnSGS3, a putative antiviral gene) with different genus viruses by constructing BnSGS3-overexpressing (BnSGS3-Ov) and BnSGS3-silenced (BnSGS3-Si) oilseed rape (cv. Zhongshuang No. 6) plants. These three viruses are Oilseed rape mosaic virus (ORMV), Turnip mosaic virus (TuMV) and Cucumber mosaic virus (CMV). The native BnSGS3 expressed in all examined tissues with the highest expression in siliques. All three viruses induced BnSGS3 expression, but ORMV induced a dramatic increase in the BnSGS3-Ov plants, followed by TuMV and CMV. Upon inoculation with three different viruses, transcript abundance of BnSGS3 gene follows: BnSGS3-Ov > non-transgenic plants > BnSGS3-Si. The accumulation quantities of ORMV and TuMV exhibited a similar trend. However, CMV accumulation showed an opposite trend where virus accumulations were negatively correlated with BnSGS3 expression. The results suggest that BnSGS3 selectively inhibits CMV accumulation but promotes ORMV and TuMV accumulation. BnSGS3 should be used in different ways (up- and down-regulation) for breeding virus-resistant oilseed rape varieties. PMID:26225990

  3. BnSGS3 Has Differential Effects on the Accumulation of CMV, ORMV and TuMV in Oilseed Rape

    PubMed Central

    Chen, Quan; Wang, Jie; Hou, Mingsheng; Liu, Shengyi; Huang, Junyan; Cai, Li

    2015-01-01

    Virus diseases greatly affect oilseed rape (Brassica napus) production. Investigating antiviral genes may lead to the development of disease-resistant varieties of oilseed rape. In this study, we examined the effects of the suppressor of gene silencing 3 in Brassica napus (BnSGS3, a putative antiviral gene) with different genus viruses by constructing BnSGS3-overexpressing (BnSGS3-Ov) and BnSGS3-silenced (BnSGS3-Si) oilseed rape (cv. Zhongshuang No. 6) plants. These three viruses are Oilseed rape mosaic virus (ORMV), Turnip mosaic virus (TuMV) and Cucumber mosaic virus (CMV). The native BnSGS3 expressed in all examined tissues with the highest expression in siliques. All three viruses induced BnSGS3 expression, but ORMV induced a dramatic increase in the BnSGS3-Ov plants, followed by TuMV and CMV. Upon inoculation with three different viruses, transcript abundance of BnSGS3 gene follows: BnSGS3-Ov > non-transgenic plants > BnSGS3-Si. The accumulation quantities of ORMV and TuMV exhibited a similar trend. However, CMV accumulation showed an opposite trend where virus accumulations were negatively correlated with BnSGS3 expression. The results suggest that BnSGS3 selectively inhibits CMV accumulation but promotes ORMV and TuMV accumulation. BnSGS3 should be used in different ways (up- and down-regulation) for breeding virus-resistant oilseed rape varieties. PMID:26225990

  4. Kinesin-14 and kinesin-5 antagonistically regulate microtubule nucleation by γ-TuRC in yeast and human cells.

    PubMed

    Olmsted, Zachary T; Colliver, Andrew G; Riehlman, Timothy D; Paluh, Janet L

    2014-01-01

    Bipolar spindle assembly is a critical control point for initiation of mitosis through nucleation and organization of spindle microtubules and is regulated by kinesin-like proteins. In fission yeast, the kinesin-14 Pkl1 binds the γ-tubulin ring complex (γ-TuRC) microtubule-organizing centre at spindle poles and can alter its structure and function. Here we show that kinesin-14 blocks microtubule nucleation in yeast and reveal that this inhibition is countered by the kinesin-5 protein, Cut7. Furthermore, we demonstrate that Cut7 binding to γ-TuRC and the Cut7 BimC domain are both required for inhibition of Pkl1. We also demonstrate that a yeast kinesin-14 peptide blocks microtubule nucleation in two human breast cancer cell lines, suggesting that this mechanism is evolutionarily conserved. In conclusion, using genetic, biochemical and cell biology approaches we uncover antagonistic control of microtubule nucleation at γ-TuRC by two kinesin-like proteins, which may represent an attractive anti-mitotic target for cancer therapies. PMID:25348260

  5. A conserved P-loop anchor limits the structural dynamics that mediate nucleotide dissociation in EF-Tu

    PubMed Central

    Mercier, Evan; Girodat, Dylan; Wieden, Hans-Joachim

    2015-01-01

    The phosphate-binding loop (P-loop) is a conserved sequence motif found in mononucleotide-binding proteins. Little is known about the structural dynamics of this region and its contribution to the observed nucleotide binding properties. Understanding the underlying design principles is of great interest for biomolecular engineering applications. We have used rapid-kinetics measurements in vitro and molecular dynamics (MD) simulations in silico to investigate the relationship between GTP-binding properties and P-loop structural dynamics in the universally conserved Elongation Factor (EF) Tu. Analysis of wild type EF-Tu and variants with substitutions at positions in or adjacent to the P-loop revealed a correlation between P-loop flexibility and the entropy of activation for GTP dissociation. The same variants demonstrate more backbone flexibility in two N-terminal amino acids of the P-loop during force-induced EF-Tu·GTP dissociation in Steered Molecular Dynamics simulations. Amino acids Gly18 and His19 are involved in stabilizing the P-loop backbone via interactions with the adjacent helix C. We propose that these P-loop/helix C interactions function as a conserved P-loop anchoring module and identify the presence of P-loop anchors within several GTPases and ATPases suggesting their evolutionary conservation. PMID:25566871

  6. [Health care and disease prevention in Jiujing Tu (Illustration of moxibustion) of the Dunhuang caves].

    PubMed

    Wang, Tian-sheng; Tang, Zhi-gang; Zhang, Rui; Li, Hui-xia; Chen, Yu-hui; Lü, Lan-ping

    2011-04-01

    Figure four of the Jiujing Tu (Illustration of Moxibustion) of the Dunhuang Caves is the earliest and the most complete recording of treatment for five kinds of strain and seven kinds of impairments in the history of acupuncture and moxibustion. Figure 12 is held as a mystery since it only provided illustrations without indications. Through analysis and approved by clinical experiences, it is held that the two figures are companion illustrations on prevention and treatment of five kinds of strain and seven kinds of impairments as well as health keeping with moxibustion. The point prescriptions in these two figures are defined according to the tri-gram in Yijing (The Book of Change), which allowed the maximization of harmony between the human and the nature. Recovery and health are thus fulfilled through regulation on points at the head, trunk and four extremities of the body. And it is considered to have great significance for promoting the development of the present acupuncture and moxibustion theory since it is effective in both preventing and curing diseases caused by deficient and stagnation conditions such as the wei (flaccidity) syndrome, bi (arthralgia) syndrome, paralysis, dementia, asthma and so on. PMID:21528610

  7. TuBaFrost 1: Uniting local frozen tumour banks into a European network: an overview.

    PubMed

    Riegman, P H J; Dinjens, W N M; Oomen, M H A; Spatz, A; Ratcliffe, C; Knox, K; Mager, R; Kerr, D; Pezzella, F; van Damme, B; van de Vijver, M; van Boven, H; Morente, M M; Alonso, S; Kerjaschki, D; Pammer, J; Lopez-Guerrero, J A; Llombart Bosch, A; Carbone, A; Gloghini, A; Teodorovic, I; Isabelle, M; Jaminé, D; Passioukov, A; Lejeune, S; Therasse, P; van Veen, E-B; Lam, K H; Oosterhuis, J W

    2006-11-01

    TuBaFrost is the consortium responsible for the creation of a virtual European human frozen tumour tissue bank: a collection of high quality frozen residual, accurately classified tumour tissue samples, which are stored in European cancer centres and universities. This virtual tissue bank, searchable on the internet, has rules for access and use, and a code of conduct to comply with the various legal and ethical regulations in European countries. The easy accessibility and the European scale of the bank will result in the availability of a large number of samples even of rarer tumour types. Standardisation of collection, storage and quality control throughout the network is achieved minimising inter-institutional variability. A website providing access to upload, search and request samples is a key tool of the tissue bank. The search engine makes use of virtual microscopy. An overview of the development of the European virtual frozen tissue bank infrastructure is described in this paper. The various key aspects are described in more detail in a series of articles to appear in this Journal. PMID:17027254

  8. DeitY-TU face database: its design, multiple camera capturing, characteristics, and evaluation

    NASA Astrophysics Data System (ADS)

    Bhowmik, Mrinal Kanti; Saha, Kankan; Saha, Priya; Bhattacharjee, Debotosh

    2014-10-01

    The development of the latest face databases is providing researchers different and realistic problems that play an important role in the development of efficient algorithms for solving the difficulties during automatic recognition of human faces. This paper presents the creation of a new visual face database, named the Department of Electronics and Information Technology-Tripura University (DeitY-TU) face database. It contains face images of 524 persons belonging to different nontribes and Mongolian tribes of north-east India, with their anthropometric measurements for identification. Database images are captured within a room with controlled variations in illumination, expression, and pose along with variability in age, gender, accessories, make-up, and partial occlusion. Each image contains the combined primary challenges of face recognition, i.e., illumination, expression, and pose. This database also represents some new features: soft biometric traits such as mole, freckle, scar, etc., and facial anthropometric variations that may be helpful for researchers for biometric recognition. It also gives an equivalent study of the existing two-dimensional face image databases. The database has been tested using two baseline algorithms: linear discriminant analysis and principal component analysis, which may be used by other researchers as the control algorithm performance score.

  9. Flight Test Measurements From The Tu- 144LL Structure/Cabin Noise Experiment

    NASA Technical Reports Server (NTRS)

    Rizzi, Stephen A.; Rackl, Robert G.; Andrianov, Eduard V.

    2000-01-01

    During the period September 1997 to February 1998, the Tupolev 144 Supersonic Flyine Laboratory was used to obtain data for the purpose of enlarging the data base used by models for the prediction of cabin noise in supersonic passenger airplanes. Measured were: turbulent boundary layer pressure fluctuations on the fuselage in seven instrumented window blanks distributed over the length of the fuselage; structural response with accelerometers on skin panels close to those window blanks-, interior noise with microphones at the same fuselage bay stations as those window blanks. Flight test points were chosen to cover much of the TU- 144's flight envelope, as well as to obtain as large a unit Reynolds number range as possible at various Mach numbers: takeoff, landing, six subsonic cruise conditions, and eleven supersonic conditions up to Mach 2. Engine runups and reverberation times were measured with a stationary aircraft. The data in the form of time histories of the acoustic signals, together with auxiliary data and basic MATLAB processing modules, are available on CD-R disks.

  10. Improved four-color flow cytometry method using fluo-3 and triple immunofluorescence for analysis of intracellular calcium ion ([Ca2+]i) fluxes among mouse lymph node B- and T-lymphocyte subsets.

    PubMed

    Greimers, R; Trebak, M; Moutschen, M; Jacobs, N; Boniver, J

    1996-03-01

    A visible-light, dual-laser, flow cytometric method was developed for the simultaneous analysis of intracellular ionized calcium concentration ([Ca2+]i) and three cell-surface markers (CD4, CD8, and Thy-1.2 antigens) by using the calcium probe fluo-3 and using R-phycoerythrin (PE), peridinin chlorophyll-alpha protein (PerCP), and allophycocyanin (APC) conjugated monoclonal antibodies (MoAbs). This improved method was used in the analysis of [Ca2+]i mobilization upon in vitro stimulation with mitogenic lectins [phytohaemagglutinin (PHA) or concanavalin A (ConA)], anti-CD3 MoAbs, or A23187 calcium ionophore in the heterogeneous lymph node cell populations from healthy C57BL/Ka mice. The present results show that the calcium responses were heterogeneous and dependent on the cellular immunophenotype, not only on lectins or anti-CD3 MoAbs stimulation, but also on the receptor-independent A23187 ionophore stimulation. An in situ fluo-3 calibration method (using A23187 and metabolic poisons in Ca2+ /EGTA buffers with known free calcium concentrations) indicated a resting [Ca2+]i in lymphocytes of 103 +/- 23 nM (mean +/- S.D.) but with significant differences between the [Ca2+]i in B cells and in all of the T-cell subsets (CD4+Thy-1+, CD4+Thy-1-, and CD8+T cells). Both the B cells and the T-cell subsets showed an increase of fluo-3 fluorescence upon in vitro stimulation with ConA or PHA, but the calcium mobilization following lectin stimulation was time delayed in all T-cell subsets. Only the T cells, including the CD4+Thy-1- subset, responded to anti-CD3 MoAbs. The percentage of responding cells upon stimulation with ConA was higher in T cells than in B cells. By contrast, PHA gave a higher response in B cells. After stimulation with different mitogens, [Ca2+]i increased in both CD4+ and CD8+ T-cell subsets. However, the percentage of responding cells was far higher in the CD4+Thy-1+ subset than in the CD4+Thy-1- or the CD8+T-cell subsets. The stimulation with A23187

  11. Flood risk analysis and adaptive strategy in context of uncertainties: a case study of Nhieu Loc Thi Nghe Basin, Ho Chi Minh City

    NASA Astrophysics Data System (ADS)

    Ho, Long-Phi; Chau, Nguyen-Xuan-Quang; Nguyen, Hong-Quan

    2013-04-01

    The Nhieu Loc - Thi Nghe basin is the most important administrative and business area of Ho Chi Minh City. Due to system complexity of the basin such as the increasing trend of rainfall intensity, (tidal) water level and land subsidence, the simulation of hydrological, hydraulic variables for flooding prediction seems rather not adequate in practical projects. The basin is still highly vulnerable despite of multi-million USD investment for urban drainage improvement projects since the last decade. In this paper, an integrated system analysis in both spatial and temporal aspects based on statistical, GIS and modelling approaches has been conducted in order to: (1) Analyse risks before and after projects, (2) Foresee water-related risk under uncertainties of unfavourable driving factors and (3) Develop a sustainable flood risk management strategy for the basin. The results show that given the framework of risk analysis and adaptive strategy, certain urban developing plans in the basin must be carefully revised and/or checked in order to reduce the highly unexpected loss in the future

  12. Assessment of waterfront location in hardened concrete by GPR within COST Action TU1208

    NASA Astrophysics Data System (ADS)

    Rodríguez-Abad, Isabel; Klysz, Gilles; Balayssac, Jean Paul; Pajewski, Lara

    2016-04-01

    This work focuses on the analysis of the capability of Ground-Penetrating radar (GPR) technique for evaluating how the water penetrates into concrete samples by means of the assessment of the waterfront advance. Research activities have been carried out during a Short-Term Scientific Missions (STSMs) funded by the COST (European Cooperation in Science and Technology) Action TU1208 "Civil Engineering Applications of Ground Penetrating Radar" in November 2015. The evaluation of water penetrability is crucial in most building materials, such us concrete, since, water and aggressive chemical agents dissolved therein contribute to the deterioration of the material. A number of techniques have been developed to measure their advance in concrete. Although the most common method for measuring water content is the gravimetric method by observing the change in mass, this method has a large number of disadvantages. In this context, non-destructive techniques as GPR play an interesting role. In particular, the application of GPR in the building materials area is providing very promising and interesting results regarding the building materials characterization and especially concrete deterioration evaluation [1-3]. In addition, recent experimental studies highlight the strong relation between wave propagation parameters (velocity and energy level) and water content advance [4-5]. Water content has a decisive influence on dielectric properties and those might be assessed by the study of the wave properties that are derived by using GPR. Therefore, the waterfront advance will result in a change on wave parameters. In line with this, this research is focused on the development of specific processing algorithms necessary to understand how the water penetrates and how the wave parameters will be affected regarding the location of the antenna in reference to the water absorption direction. For this purpose, concrete samples were manufactured, which after curing (90 days) and oven

  13. Civil Engineering Applications of Ground Penetrating Radar: Research Perspectives in COST Action TU1208

    NASA Astrophysics Data System (ADS)

    Pajewski, Lara; Benedetto, Andrea; Loizos, Andreas; Slob, Evert; Tosti, Fabio

    2013-04-01

    can be used by GPR operators to identify the signatures generated by uncommon targets or by composite structures. Repeated evaluations of the electromagnetic field scattered by known targets can be performed by a forward solver, in order to estimate - through comparison with measured data - the physics and geometry of the region investigated by the GPR. It is possible to identify three main areas, in the GPR field, that have to be addressed in order to promote the use of this technology in the civil engineering. These are: a) increase of the system sensitivity to enable the usability in a wider range of conditions; b) research novel data processing algorithms/analysis tools for the interpretation of GPR results; c) contribute to the development of new standards and guidelines and to training of end users, that will also help to increase the awareness of operators. In this framework, the COST Action TU1208 "Civil Engineering Applications of Ground Penetrating Radar", proposed by Lara Pajewski, "Roma Tre" University, Rome, Italy, has been approved in November 2012 and is going to start in April 2013. It is a 4-years ambitious project already involving 17 European Countries (AT, BE, CH, CZ, DE, EL, ES, FI, FR, HR, IT, NL, NO, PL, PT, TR, UK), as well as Australia and U.S.A. The project will be developed within the frame of a unique approach based on the integrated contribution of University researchers, software developers, geophysics experts, Non-Destructive Testing equipment designers and producers, end users from private companies and public agencies. The main objective of the COST Action TU1208 is to exchange and increase scientific-technical knowledge and experience of GPR techniques in civil engineering, whilst promoting the effective use of this safe and non-destructive technique in the monitoring of systems. In this interdisciplinary Action, advantages and limitations of GPR will be highlighted, leading to the identification of gaps in knowledge and technology

  14. COST Action TU1208 "Civil Engineering Applications of Ground Penetrating Radar:" ongoing research activities and mid-term results

    NASA Astrophysics Data System (ADS)

    Pajewski, Lara; Benedetto, Andrea; Loizos, Andreas; Slob, Evert; Tosti, Fabio

    2015-04-01

    This work aims at presenting the ongoing activities and mid-term results of the COST (European COoperation in Science and Technology) Action TU1208 'Civil Engineering Applications of Ground Penetrating Radar.' Almost three hundreds experts are participating to the Action, from 28 COST Countries (Austria, Belgium, Croatia, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Ireland, Italy, Latvia, Malta, Macedonia, The Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey, United Kingdom), and from Albania, Armenia, Australia, Egypt, Hong Kong, Jordan, Israel, Philippines, Russia, Rwanda, Ukraine, and United States of America. In September 2014, TU1208 has been praised among the running Actions as 'COST Success Story' ('The Cities of Tomorrow: The Challenges of Horizon 2020,' September 17-19, 2014, Torino, IT - A COST strategic workshop on the development and needs of the European cities). The principal goal of the COST Action TU1208 is to exchange and increase scientific-technical knowledge and experience of GPR techniques in civil engineering, whilst simultaneously promoting throughout Europe the effective use of this safe and non-destructive technique in the monitoring of infrastructures and structures. Moreover, the Action is oriented to the following specific objectives and expected deliverables: (i) coordinating European scientists to highlight problems, merits and limits of current GPR systems; (ii) developing innovative protocols and guidelines, which will be published in a handbook and constitute a basis for European standards, for an effective GPR application in civil- engineering tasks; safety, economic and financial criteria will be integrated within the protocols; (iii) integrating competences for the improvement and merging of electromagnetic scattering techniques and of data- processing techniques; this will lead to a novel freeware tool for the localization of buried objects

  15. COST Action TU1208 "Civil Engineering Applications of Ground Penetrating Radar": ongoing research activities and third-year results

    NASA Astrophysics Data System (ADS)

    Pajewski, Lara; Benedetto, Andrea; Loizos, Andreas; Tosti, Fabio

    2016-04-01

    This work aims at disseminating the ongoing research activities and third-year results of the COST (European COoperation in Science and Technology) Action TU1208 "Civil Engineering Applications of Ground Penetrating Radar." About 350 experts are participating to the Action, from 28 COST Countries (Austria, Belgium, Croatia, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Ireland, Italy, Latvia, Malta, Macedonia, The Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey, United Kingdom), and from Albania, Armenia, Australia, Colombia, Egypt, Hong Kong, Jordan, Israel, Philippines, Russia, Rwanda, Ukraine, and United States of America. In September 2014, TU1208 has been recognised among the running Actions as "COST Success Story" ("The Cities of Tomorrow: The Challenges of Horizon 2020," September 17-19, 2014, Torino, IT - A COST strategic workshop on the development and needs of the European cities). The principal goal of the COST Action TU1208 is to exchange and increase scientific-technical knowledge and experience of GPR techniques in civil engineering, whilst simultaneously promoting throughout Europe the effective use of this safe and non-destructive technique in the monitoring of infrastructures and structures. Moreover, the Action is oriented to the following specific objectives and expected deliverables: (i) coordinating European scientists to highlight problems, merits and limits of current GPR systems; (ii) developing innovative protocols and guidelines, which will be published in a handbook and constitute a basis for European standards, for an effective GPR application in civil- engineering tasks; safety, economic and financial criteria will be integrated within the protocols; (iii) integrating competences for the improvement and merging of electromagnetic scattering techniques and of data- processing techniques; this will lead to a novel freeware tool for the localization of

  16. Overview and comparative study of GPR international standards and guidelines - COST Action TU1208

    NASA Astrophysics Data System (ADS)

    Pajewski, Lara; Marciniak, Marian; Benedetto, Andrea; Tosti, Fabio

    2016-04-01

    Ground Penetrating Radar (GPR) can be effectively used for non-destructive testing of composite structures and diagnostics affecting the whole life-cycle of civil engineering works. Nevertheless, few recognised international standards exist in this field and inhomogeneous recommendations are present in different countries. Moreover, the levels of knowledge, awareness and experience regarding the use of GPR in civil engineering vary strongly across different European areas. The COST Action TU1208 is working hard on leveraging these differences, by sharing and disseminating knowledge and experience, as well as by developing guidelines and protocols for a safe and effective use of GPR in civil engineering. GPR users need to know which is the best way to conduct GPR measurements and what the quality level for the results should be. The TU1208 guidelines will ensure a higher efficiency and quality of GPR services and they will constitute a scientific basis for the introduction of European Standards on the application of GPR in civil engineering. The aim of this contribution is to present an in-depth overview and critical analysis of the existing GPR international and national standards and guidelines. The main documents considered in our work are listed and briefly described in the following. Three standards are provided by the American Society for Testing and Materials (ASTM), to guide the GPR use for subsurface investigation, evaluation of asphalt-covered concrete bridge decks, and determination of pavement-layer thickness: 1. ASTM D6432-11, Standard Guide for Using the Surface Ground Penetrating Radar Method for Subsurface Investigation, ASTM International, West Conshohocken, PA, 2011, www.astm.org, DOI: 10.1520/D6432-11. 2. ASTM D6087-08, Standard Test Method for Evaluating Asphalt-Covered Concrete Bridge Decks Using Ground Penetrating Radar, ASTM International, West Conshohocken, PA, 2008, www.astm.org, DOI: 10.1520/D6087-08. 3. ASTM D4748-10, Standard Test Method

  17. An Entamoeba histolytica ADP-ribosyl transferase from the diphtheria toxin family modifies the bacterial elongation factor Tu.

    PubMed

    Avila, Eva E; Rodriguez, Orlando I; Marquez, Jaqueline A; Berghuis, Albert M

    2016-06-01

    ADP-ribosyl transferases are enzymes involved in the post-translational modification of proteins; they participate in multiple physiological processes, pathogenesis and host-pathogen interactions. Several reports have characterized the functions of these enzymes in viruses, prokaryotes and higher eukaryotes, but few studies have reported ADP-ribosyl transferases in lower eukaryotes, such as parasites. The locus EHI_155600 from Entamoeba histolytica encodes a hypothetical protein that possesses a domain from the ADP-ribosylation superfamily; this protein belongs to the diphtheria toxin family according to a homology model using poly-ADP-ribosyl polymerase 12 (PARP12 or ARTD12) as a template. The recombinant protein expressed in Escherichia coli exhibited in vitro ADP-ribosylation activity that was dependent on the time and temperature. Unlabeled βNAD(+), but not ADP-ribose, competed in the enzymatic reaction using biotin-βNAD(+) as the ADP-ribose donor. The recombinant enzyme, denominated EhToxin-like, auto-ADP-ribosylated and modified an acceptor from E. coli that was identified by MS/MS as the elongation factor Tu (EF-Tu). To the best of our knowledge, this is the first report to identify an ADP-ribosyl transferase from the diphtheria toxin family in a protozoan parasite. The known toxins from this family (i.e., the diphtheria toxin, the Pseudomonas aeruginosa toxin Exo-A, and Cholix from Vibrio cholerae) modify eukaryotic elongation factor two (eEF-2), whereas the amoeba EhToxin-like modified EF-Tu, which is another elongation factor involved in protein synthesis in bacteria and mitochondria. PMID:27234208

  18. COST Action TU1208 "Civil Engineering Applications of Ground Penetrating Radar:" ongoing research activities and mid-term results

    NASA Astrophysics Data System (ADS)

    Pajewski, Lara; Benedetto, Andrea; Loizos, Andreas; Slob, Evert; Tosti, Fabio

    2015-04-01

    This work aims at presenting the ongoing activities and mid-term results of the COST (European COoperation in Science and Technology) Action TU1208 'Civil Engineering Applications of Ground Penetrating Radar.' Almost three hundreds experts are participating to the Action, from 28 COST Countries (Austria, Belgium, Croatia, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Ireland, Italy, Latvia, Malta, Macedonia, The Netherlands, Norway, Poland, Portugal, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey, United Kingdom), and from Albania, Armenia, Australia, Egypt, Hong Kong, Jordan, Israel, Philippines, Russia, Rwanda, Ukraine, and United States of America. In September 2014, TU1208 has been praised among the running Actions as 'COST Success Story' ('The Cities of Tomorrow: The Challenges of Horizon 2020,' September 17-19, 2014, Torino, IT - A COST strategic workshop on the development and needs of the European cities). The principal goal of the COST Action TU1208 is to exchange and increase scientific-technical knowledge and experience of GPR techniques in civil engineering, whilst simultaneously promoting throughout Europe the effective use of this safe and non-destructive technique in the monitoring of infrastructures and structures. Moreover, the Action is oriented to the following specific objectives and expected deliverables: (i) coordinating European scientists to highlight problems, merits and limits of current GPR systems; (ii) developing innovative protocols and guidelines, which will be published in a handbook and constitute a basis for European standards, for an effective GPR application in civil- engineering tasks; safety, economic and financial criteria will be integrated within the protocols; (iii) integrating competences for the improvement and merging of electromagnetic scattering techniques and of data- processing techniques; this will lead to a novel freeware tool for the localization of buried objects

  19. The SCID-hu mouse as a tool in immunotoxicological risk assessment: effects of 2-acetyl-4(5)-tetrahydroxybutyl-imidazole (THI) and di-n-butyltin dichloride (DBTC) on the human thymus in SCID-hu mice.

    PubMed

    de Heer, C; Schuurman, H J; Houben, G F; Pieters, R H; Penninks, A H; van Loveren, H

    1995-06-26

    SCID mice engrafted with human fetal thymus and liver tissue fragments (SCID-hu mice) are currently considered as a new tool in human immunotoxicological risk assessment. Testing of various immunotoxicants exerting thymotoxicity via different intrathymic target cell types is necessary for validation of this model. Therefore, SCID-hu mice were exposed to 2-acetyl-4(5)-(1,2,3,4-tetrahydroxybutyl)-imidazole (THI), the immunotoxic component in the food additive, Caramel Colour III, or the organotin compound, di-n-butyltin dichloride (DBTC). Histopathological examination of the human thymus grafts of SCID-hu mice either exposed to THI or to DBTC showed a reduction in the relative size of the thymus cortex, an effect also described in rodents. These results indicate that the human thymus is a target for the immunotoxic action of both THI and DBTC. In addition, they indicate the promising potential of the SCID-hu mouse model as a tool for human immunotoxicological risk assessment. PMID:7624878

  20. Neutron Knockout to Probe 3N Forces in the Ca Isotopes

    NASA Astrophysics Data System (ADS)

    Crawford, Heather; NSCL Experiment e12029 Collaboration

    2013-10-01

    Recent calculations by Holt et al. have suggested that the inclusion of 3N forces to describe the structure of neutron-rich Ca isotopes provides a more realistic description of the nuclear structure. Mass measurements have confirmed the importance of including 3N forces compared to NN-only interactions, but cannot discriminate between the predictions of phenomenological interactions and the NN+3N microscopic calculations. Neutron knockout along the Z = 20 isotopes provides an opportunity to test the results of NN+3N calculations against phenomenological interactions. The calculations of Holt et al. predict a fragmentation of the 1f7 / 2 neutron strength from the first 7/2- state in 49Ca into higher lying states, in contrast to the predictions of both GXPF1 and KB3G which concentrate the strength in the lowest lying 7/2- state. Differences are also observed in the summed f7 / 2 strength to bound nuclear states in both 50Ca and 49Ca neutron knockout. We will report on a systematic study of one-neutron knockout along the Ca isotopes using GRETINA+S800. Comparison of experimental spectroscopic factors to calculations will provide insight into the role of 3N forces in describing the Z = 20 isotopes. With collaborators from LBNL, NSCL/MSU, ANL, Central Michigan University, TRIUMF/UBC and TU-Darmstadt.

  1. Elongation Factor Tu and Heat Shock Protein 70 Are Membrane-Associated Proteins from Mycoplasma ovipneumoniae Capable of Inducing Strong Immune Response in Mice.

    PubMed

    Jiang, Fei; He, Jinyan; Navarro-Alvarez, Nalu; Xu, Jian; Li, Xia; Li, Peng; Wu, Wenxue

    2016-01-01

    Chronic non-progressive pneumonia, a disease that has become a worldwide epidemic has caused considerable loss to sheep industry. Mycoplasma ovipneumoniae (M. ovipneumoniae) is the causative agent of interstitial pneumonia in sheep, goat and bighorn. We here have identified by immunogold and immunoblotting that elongation factor Tu (EF-Tu) and heat shock protein 70 (HSP 70) are membrane-associated proteins on M. ovipneumonaiea. We have evaluated the humoral and cellular immune responses in vivo by immunizing BALB/c mice with both purified recombinant proteins rEF-Tu and rHSP70. The sera of both rEF-Tu and rHSP70 treated BALB/c mice demonstrated increased levels of IgG, IFN-γ, TNF-α, IL-12(p70), IL-4, IL-5 and IL-6. In addition, ELISPOT assay showed significant increase in IFN-γ+ secreting lymphocytes in the rHSP70 group when compared to other groups. Collectively our study reveals that rHSP70 induces a significantly better cellular immune response in mice, and may act as a Th1 cytokine-like adjuvant in immune response induction. Finally, growth inhibition test (GIT) of M. ovipneumoniae strain Y98 showed that sera from rHSP70 or rEF-Tu-immunized mice inhibited in vitro growth of M. ovipneumoniae. Our data strongly suggest that EF-Tu and HSP70 of M. ovipneumoniae are membrane-associated proteins capable of inducing antibody production, and cytokine secretion. Therefore, these two proteins may be potential candidates for vaccine development against M. ovipneumoniae infection in sheep. PMID:27537186

  2. Construction of a genetic map based on high-throughput SNP genotyping and genetic mapping of a TuMV resistance locus in Brassica rapa.

    PubMed

    Chung, Hee; Jeong, Young-Min; Mun, Jeong-Hwan; Lee, Soo-Seong; Chung, Won-Hyong; Yu, Hee-Ju

    2014-04-01

    Brassica rapa is a member of the Brassicaceae family and includes vegetables and oil crops that are cultivated worldwide. The introduction of durable resistance against turnip mosaic virus (TuMV) into agronomically important cultivars has been a significant challenge for genetic and horticultural breeding studies of B. rapa. Based on our previous genome-wide analysis of DNA polymorphisms between the TuMV-resistant doubled haploid (DH) line VC40 and the TuMV-susceptible DH line SR5, we constructed a core genetic map of the VCS-13M DH population, which is composed of 83 individuals derived from microspore cultures of a F1 cross between VC40 and SR5, by analyzing the segregation of 314 sequence-characterized genetic markers. The genetic markers correspond to 221 SNPs and 31 InDels of genes as well as 62 SSRs, covering 1,115.9 cM with an average distance of 3.6 cM between the adjacent marker loci. The alignment and orientation of the constructed map showed good agreement with the draft genome sequence of Chiifu, thus providing an efficient strategy to map genic sequences. Using the genetic map, a novel dominant TuMV resistance locus (TuMV-R) in the VCS-13M DH population was identified as a 0.34 Mb region in the short arm of chromosome A6 in which four CC-NBS-LRR resistance genes and two pathogenesis-related-1 genes reside. The genetic map developed in this study can play an important role in the genetic study of TuMV resistance and the molecular breeding of B. rapa. PMID:24326528

  3. Structure of the Acinetobacter baumannii Dithiol Oxidase DsbA Bound to Elongation Factor EF-Tu Reveals a Novel Protein Interaction Site

    PubMed Central

    Premkumar, Lakshmanane; Kurth, Fabian; Duprez, Wilko; Grøftehauge, Morten K.; King, Gordon J.; Halili, Maria A.; Heras, Begoña; Martin, Jennifer L.

    2014-01-01

    The multidrug resistant bacterium Acinetobacter baumannii is a significant cause of nosocomial infection. Biofilm formation, that requires both disulfide bond forming and chaperone-usher pathways, is a major virulence trait in this bacterium. Our biochemical characterizations show that the periplasmic A. baumannii DsbA (AbDsbA) enzyme has an oxidizing redox potential and dithiol oxidase activity. We found an unexpected non-covalent interaction between AbDsbA and the highly conserved prokaryotic elongation factor, EF-Tu. EF-Tu is a cytoplasmic protein but has been localized extracellularly in many bacterial pathogens. The crystal structure of this complex revealed that the EF-Tu switch I region binds to the non-catalytic surface of AbDsbA. Although the physiological and pathological significance of a DsbA/EF-Tu association is unknown, peptides derived from the EF-Tu switch I region bound to AbDsbA with submicromolar affinity. We also identified a seven-residue DsbB-derived peptide that bound to AbDsbA with low micromolar affinity. Further characterization confirmed that the EF-Tu- and DsbB-derived peptides bind at two distinct sites. These data point to the possibility that the non-catalytic surface of DsbA is a potential substrate or regulatory protein interaction site. The two peptides identified in this work together with the newly characterized interaction site provide a novel starting point for inhibitor design targeting AbDsbA. PMID:24860094

  4. Elongation Factor Tu and Heat Shock Protein 70 Are Membrane-Associated Proteins from Mycoplasma ovipneumoniae Capable of Inducing Strong Immune Response in Mice

    PubMed Central

    Jiang, Fei; He, Jinyan; Navarro-Alvarez, Nalu; Xu, Jian; Li, Xia; Li, Peng; Wu, Wenxue

    2016-01-01

    Chronic non-progressive pneumonia, a disease that has become a worldwide epidemic has caused considerable loss to sheep industry. Mycoplasma ovipneumoniae (M. ovipneumoniae) is the causative agent of interstitial pneumonia in sheep, goat and bighorn. We here have identified by immunogold and immunoblotting that elongation factor Tu (EF-Tu) and heat shock protein 70 (HSP 70) are membrane-associated proteins on M. ovipneumonaiea. We have evaluated the humoral and cellular immune responses in vivo by immunizing BALB/c mice with both purified recombinant proteins rEF-Tu and rHSP70. The sera of both rEF-Tu and rHSP70 treated BALB/c mice demonstrated increased levels of IgG, IFN-γ, TNF-α, IL-12(p70), IL-4, IL-5 and IL-6. In addition, ELISPOT assay showed significant increase in IFN-γ+ secreting lymphocytes in the rHSP70 group when compared to other groups. Collectively our study reveals that rHSP70 induces a significantly better cellular immune response in mice, and may act as a Th1 cytokine-like adjuvant in immune response induction. Finally, growth inhibition test (GIT) of M. ovipneumoniae strain Y98 showed that sera from rHSP70 or rEF-Tu-immunized mice inhibited in vitro growth of M. ovipneumoniae. Our data strongly suggest that EF-Tu and HSP70 of M. ovipneumoniae are membrane-associated proteins capable of inducing antibody production, and cytokine secretion. Therefore, these two proteins may be potential candidates for vaccine development against M. ovipneumoniae infection in sheep. PMID:27537186

  5. Cloning of Genes Coding for the Three Subunits of Thiocyanate Hydrolase of Thiobacillus thioparus THI 115 and Their Evolutionary Relationships to Nitrile Hydratase

    PubMed Central

    Katayama, Yoko; Matsushita, Yasuhiko; Kaneko, Miyuki; Kondo, Mai; Mizuno, Tadayoshi; Nyunoya, Hiroshi

    1998-01-01

    Thiocyanate hydrolase is a newly found enzyme from Thiobacillus thioparus THI 115 that converts thiocyanate to carbonyl sulfide and ammonia (Y. Katayama, Y. Narahara, Y. Inoue, F. Amano, T. Kanagawa, and H. Kuraishi, J. Biol. Chem. 267:9170–9175, 1992). We have cloned and sequenced the scn genes that encode the three subunits of the enzyme. The scnB, scnA, and scnC genes, arrayed in this order, contained open reading frames encoding sequences of 157, 126, and 243 amino acid residues, respectively, for the β, α, and γ subunits, respectively. Each open reading frame was preceded by a typical Shine-Dalgarno sequence. The deduced amino-terminal peptide sequences for the three subunits were in fair agreement with the chemically determined sequences. The protein molecular mass calculated for each subunit was compatible with that determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. From a computer analysis, thiocyanate hydrolase showed significant homologies to bacterial nitrile hydratases known to convert nitrile to the corresponding amide, which is further hydrolyzed by amidase to form acid and ammonia. The two enzymes were homologous over regions corresponding to almost the entire coding regions of the genes: the β and α subunits of thiocyanate hydrolase were homologous to the amino- and carboxyl-terminal halves of the β subunit of nitrile hydratase, and the γ subunit of thiocyanate hydrolase was homologous to the α subunit of nitrile hydratase. Comparisons of the catalytic properties of the two homologous enzymes support the model for the reaction steps of thiocyanate hydrolase that was previously presented on the basis of biochemical analyses. PMID:9573140

  6. CD90/THY1 is over-expressed in prostate cancer-associated fibroblasts and could serve as a cancer biomarker

    SciTech Connect

    True, Lawrence D.; Zhang, Hui; Ye, Mingliang; Huang, Chung-Ying; Nelson, Peter S.; Von Haller, Priska D.; Tjoelker, Larry W.; Kim, Jong Seo; Qian, Weijun; Smith, Richard D.; Ellis, William J.; Liebeskind, Emily S.; Liu, Alvin Y.

    2010-10-01

    A by-product in the processing of prostate tissue for cell sorting by collagenase digestion is the media supernatant that remains after the cells are harvested. These supernatants contain proteins made by the cells within the tissue. Quantitative proteomic analysis of Nglycosylated proteins detected an increased amount of CD90/THY1 in cancer supernatants compared to non-cancer supernatants. Immunohistochemistry showed that in all carcinomas, regardless of Gleason grade, a layer of CD90-positive stromal fibroblastic cells, approximately 5-to-10 cells deep, was localized to tumor glands. In contrast, a no more than 1-cell wide girth of CD90-positive stromal cells was found around benign glands. The increased number of CD90-positive stromal cells in cancer correlated with overexpression of CD90 mRNA detected by gene expression analysis of stromal cells obtained by laser-capture microdissection. There is increasing evidence that cancer-associated stroma plays a role in both tumor progression and carcinogenesis. Most experiments to identify cancer biomarkers have focused on the cancer cells. CD90, being a marker for prostate cancer-associated stroma, might be a potential biomarker for this cancer. A non-invasive test could be provided by a urine test. Proteomic analysis of urine from patients with prostate cancer identified CD90; conversely, CD90 was not detected in the urine of post-prostatectomy patients. Furthermore, this urinary CD90 protein was a variant CD90 protein not known to be expressed by such cells as lymphocytes that express CD90. These CD90 results were obtained from ~90 cases consisting of proteomic analysis of tissue and urine, immunohistochemistry, Western blot analysis of tissue media, flow cytometry of cells from digested tissue, and reverse transcriptase polymerase chain reaction analysis of isolated stromal cells.

  7. Polymorphism in dhfr/dhps genes, parasite density and ex vivo response to pyrimethamine in Plasmodium falciparum malaria parasites in Thies, Senegal.

    PubMed

    Ndiaye, Daouda; Dieye, Baba; Ndiaye, Yaye D; Van Tyne, Daria; Daniels, Rachel; Bei, Amy K; Mbaye, Aminata; Valim, Clarissa; Lukens, Amanda; Mboup, Souleymane; Ndir, Omar; Wirth, Dyann F; Volkman, Sarah

    2013-12-01

    Resistance to sulfadoxine-pyrimethamine (SP) in Plasmodium falciparum malaria parasites is associated with mutations in the dihydrofolate reductase (dhfr) and dihydropteroate synthase (dhps) genes, and these mutations have spread resistance worldwide. SP, used for several years in Senegal, has been recommended for intermittent preventive treatment for malaria in pregnancy (IPTp) and has been widely implemented since 2003 in this country. There is currently limited data on SP resistance from molecular marker genotyping, and no data on pyrimethamine ex vivo sensitivity in Senegal. Molecular markers of SP resistance and pyrimethamine ex vivo sensitivity were investigated in 416 parasite samples collected from the general population, from the Thies region between 2003 and 2011. The prevalence of the N51I/C59R/S108N triple mutation in dhfr increased from 40% in 2003 to 93% in 2011. Furthermore, the prevalence of the dhfr N51I/C59R/S108N and dhps A437G quadruple mutation increased, from 20% to 66% over the same time frame, then down to 44% by 2011. There was a significant increase in the prevalence of the dhfr triple mutation, as well as an association between dhfr genotypes and pyrimethamine response. Conversely, dhps mutations in codons 436 and 437 did not show consistent variation between 2003 and 2011. These findings suggest that regular screening for molecular markers of antifolate resistance and ex vivo drug response monitoring should be incorporated with ongoing in vivo efficacy monitoring in areas where IPTp-SP is implemented and where pyrimethamine and sulfa drugs are still widely administered in the general population. PMID:24533303

  8. Bacterial Contamination in Intensive Care Unit at Al-Imam Al-Hussein Hospital in Thi-qar Province in Iraq

    PubMed Central

    Nasser, Nazar Edward; Abbas, Ali Taher; Hamed, Saad L.

    2013-01-01

    Cross- infection from patient to patient or from hospital personnel to patients represents constant hazards. It is one of the most important causes of morbidity and mortality especially in Intensive Care Unit all over the world. To identify the types and the source of bacterial contamination in ICU and to study the sensitivity of bacterial isolates to commonly used antibiotics in hospitals this study had been conducted in Al-Imam Al-Hussein hospital in Thi-qar province for the period from the 1st of September to the end of December 2011. A total of 320 swabs and samples were collected from 17 different sites of Intensive Care Unit environment and inoculated on a normal cultural media, then incubated at 37°C for 24 hour. The obtained growth revealed different bacterial colonies which had been tested for their morphological and biochemical characteristics. Sixty eight of pure isolates were obtained including 24 (35.29%) Gram positive bacterial isolates, and 44(64.71%) of Gram negative bacterial isolates, the highest rates (19.11%) of bacterial contamination had been found on the walls and the floor. Sensitivity tests for all isolates were done using 25 types of commonly used antibiotics in Iraq, among Gram negative bacteria and gram positive bacteria the genus Enterobacter spp and Staphylococcus spp respectively, showed the highest resistance to most of the tested antibiotics, MIC tests for 5 types of antibiotics being applied for the most resistant and the most sensitive isolates had identified that all isolates have a low rate of MIC against Ciprofloxacine. Bacillus spp and Enterobacter spp were the most prevalent bacterial contaminants of Intensive Care Unit environment. such contamination could be managed mostly by strict application of sterilization measures. PMID:23283046

  9. Nondestructive tests for railway monitoring. European Experience in COST Action TU1208

    NASA Astrophysics Data System (ADS)

    Fontul, Simona; Solla, Mercedes; Loizos, Andreas

    2016-04-01

    The railway monitoring is an important issue for a proper maintenance planning. With the increase in loads and travel speed, it is important to be able to diagnose the track defects and to plan the proper maintenance without interfering with the users. Traditionally, the maintenance actions are planned based on the geometric level parameters assessed without contact with the line, at traffic speed, by dedicated inspection vehicles. Nevertheless, the geometric condition of the line does not provide information on the defects causes. In order to complements the information on the causes, geophysics measurements can be performed in a nondestructive way. Among these later methods, Ground Penetrating Radar (GPR) is a quick and effective technique to evaluate infrastructure condition in a continuous manner, replacing or reducing the use of traditional drilling method. GPR application to railways infrastructures, during construction and monitoring phase, is relatively recent. It is based on the measuring of layers thicknesses and detection of structural changes. It also enables the assessment of materials properties that constitute the infrastructure and the evaluation of the different types of defects such as ballast pockets, fouled ballast, poor drainage, subgrade settlement and transitions problems. These deteriorations are generally the causes of vertical deviations in track geometry. Moreover, the development of new GPR systems with higher antenna frequencies, better data acquisition systems, more user friendly software and new algorithms for calculation of materials properties can lead to a regular use of GPR. A resume of the European experience in COST Action TU1208 of the application of GPR for railway monitoring and the measurement interpretation is presented in this paper. Also complementary nondestructive tests and other geophysical methods are referred, together with case studies of their application. The main troubleshooting and the needs for data analysis

  10. The Dynamic Atmospheres of Mira Stars: Comparing the CODEX Models to PTI Time Series Observations of TU Andromedae

    NASA Astrophysics Data System (ADS)

    Hillen, M.; Verhoelst, T.; Degroote, P.; Acke, B.; Van Winckel, H.

    2015-08-01

    We present our already-published evaluation of the effectiveness of the CODEX models, released in 2011, in representing the atmospheres of M-type Mira variables. We present a high-precision interferometric K-band time series of TU And, consisting of 50 nights that cover eight consecutive pulsation cycles. At each phase, the flux at 2.2μm was obtained, along with the spectral shape and visibility points in five channels across the K band. We show a comparison between these data and the dynamical self-excited CODEX model which gives the closest match in stellar parameters yet available. Both the spectrum and the visibilities are consistently reproduced around visual minimum phases. Near the maximum phases, however, the current models predict a photosphere that is too hot and compact, surrounded by an extended atmosphere that lacks H2O opacity, compared to the observations. A better coverage in the model parameter space is needed to make firm conclusions as to the cause of the discrepancies. In the case of TU And, the discrepancy might be lifted by adopting a lower value of the mixing length parameter combined with an increased stellar mass and/or a decreased metallicity.

  11. Overview and comparative study of GPR international standards and guidelines - COST Action TU1208

    NASA Astrophysics Data System (ADS)

    Pajewski, Lara; Marciniak, Marian; Benedetto, Andrea; Tosti, Fabio

    2016-04-01

    Ground Penetrating Radar (GPR) can be effectively used for non-destructive testing of composite structures and diagnostics affecting the whole life-cycle of civil engineering works. Nevertheless, few recognised international standards exist in this field and inhomogeneous recommendations are present in different countries. Moreover, the levels of knowledge, awareness and experience regarding the use of GPR in civil engineering vary strongly across different European areas. The COST Action TU1208 is working hard on leveraging these differences, by sharing and disseminating knowledge and experience, as well as by developing guidelines and protocols for a safe and effective use of GPR in civil engineering. GPR users need to know which is the best way to conduct GPR measurements and what the quality level for the results should be. The TU1208 guidelines will ensure a higher efficiency and quality of GPR services and they will constitute a scientific basis for the introduction of European Standards on the application of GPR in civil engineering. The aim of this contribution is to present an in-depth overview and critical analysis of the existing GPR international and national standards and guidelines. The main documents considered in our work are listed and briefly described in the following. Three standards are provided by the American Society for Testing and Materials (ASTM), to guide the GPR use for subsurface investigation, evaluation of asphalt-covered concrete bridge decks, and determination of pavement-layer thickness: 1. ASTM D6432-11, Standard Guide for Using the Surface Ground Penetrating Radar Method for Subsurface Investigation, ASTM International, West Conshohocken, PA, 2011, www.astm.org, DOI: 10.1520/D6432-11. 2. ASTM D6087-08, Standard Test Method for Evaluating Asphalt-Covered Concrete Bridge Decks Using Ground Penetrating Radar, ASTM International, West Conshohocken, PA, 2008, www.astm.org, DOI: 10.1520/D6087-08. 3. ASTM D4748-10, Standard Test Method

  12. Moisture evaluation of wood material using GPR with WARR method - COST Action TU1208

    NASA Astrophysics Data System (ADS)

    Reci, Hamza; Sbart'i, Zoubir Mehdi; Pajewski, Lara; Marciniak, Marian

    2016-04-01

    This work deals with the study of the sensitivity of GPR electromagnetic waves to moisture variation in wood material in relation with the direction of fibers and polarization of Electromagnetic field. The relations between relative permittivity and moisture content and the amplitude attenuation with distance was a target study using the direct waves in Wide Angle Radar Reflection (WARR) configuration. Comparison of results measured with reflected waves and direct waves was of main importance since they have different behavior in relation with moisture variation, due to different path of propagation. This research activity has been carried out during one Short-Term Scientific Missions (STSM) funded by the COST (European Cooperation in Science and Technology) Action TU1208 "Civil Engineering Applications of Ground Penetrating Radar" in November-December 2015. In context of durability evaluation of construction materials, several studies have been carried out by the I2M team, University of Bordeaux, using direct and reflected waves for the evaluation of water content on concrete and wood materials [1-3]. As related to the wood material there is one study carried out using the reflected waves on wood for different humidity and different wood samples, in all the direction of polarization using GPR technique ground coupled antenna at 1.5 GHz [3]. This work continued with different moisture content in order to study the behavior of direct waves as function of moisture. Results taken from those measurements are compared with them from Fixed Offset (reflected method) with one antenna (1.5GHz or 2.6GHz), realized from the previous studies from the I2M and already published [1-3]. The results taken from this work from the reflected waves, show that the effect of wood anisotropy is significant on the variation of relative permittivity with moisture content on wood sample and that is in good agreement with the previous results [3-6]. As related to the direct waves, a small

  13. Moisture evaluation of wood material using GPR with WARR method - COST Action TU1208

    NASA Astrophysics Data System (ADS)

    Reci, Hamza; Sbart'i, Zoubir Mehdi; Pajewski, Lara; Marciniak, Marian

    2016-04-01

    This work deals with the study of the sensitivity of GPR electromagnetic waves to moisture variation in wood material in relation with the direction of fibers and polarization of Electromagnetic field. The relations between relative permittivity and moisture content and the amplitude attenuation with distance was a target study using the direct waves in Wide Angle Radar Reflection (WARR) configuration. Comparison of results measured with reflected waves and direct waves was of main importance since they have different behavior in relation with moisture variation, due to different path of propagation. This research activity has been carried out during one Short-Term Scientific Missions (STSM) funded by the COST (European Cooperation in Science and Technology) Action TU1208 "Civil Engineering Applications of Ground Penetrating Radar" in November-December 2015. In context of durability evaluation of construction materials, several studies have been carried out by the I2M team, University of Bordeaux, using direct and reflected waves for the evaluation of water content on concrete and wood materials [1-3]. As related to the wood material there is one study carried out using the reflected waves on wood for different humidity and different wood samples, in all the direction of polarization using GPR technique ground coupled antenna at 1.5 GHz [3]. This work continued with different moisture content in order to study the behavior of direct waves as function of moisture. Results taken from those measurements are compared with them from Fixed Offset (reflected method) with one antenna (1.5GHz or 2.6GHz), realized from the previous studies from the I2M and already published [1-3]. The results taken from this work from the reflected waves, show that the effect of wood anisotropy is significant on the variation of relative permittivity with moisture content on wood sample and that is in good agreement with the previous results [3-6]. As related to the direct waves, a small

  14. Assessment of waterfront location in hardened concrete by GPR within COST Action TU1208

    NASA Astrophysics Data System (ADS)

    Rodríguez-Abad, Isabel; Klysz, Gilles; Balayssac, Jean Paul; Pajewski, Lara

    2016-04-01

    This work focuses on the analysis of the capability of Ground-Penetrating radar (GPR) technique for evaluating how the water penetrates into concrete samples by means of the assessment of the waterfront advance. Research activities have been carried out during a Short-Term Scientific Missions (STSMs) funded by the COST (European Cooperation in Science and Technology) Action TU1208 "Civil Engineering Applications of Ground Penetrating Radar" in November 2015. The evaluation of water penetrability is crucial in most building materials, such us concrete, since, water and aggressive chemical agents dissolved therein contribute to the deterioration of the material. A number of techniques have been developed to measure their advance in concrete. Although the most common method for measuring water content is the gravimetric method by observing the change in mass, this method has a large number of disadvantages. In this context, non-destructive techniques as GPR play an interesting role. In particular, the application of GPR in the building materials area is providing very promising and interesting results regarding the building materials characterization and especially concrete deterioration evaluation [1-3]. In addition, recent experimental studies highlight the strong relation between wave propagation parameters (velocity and energy level) and water content advance [4-5]. Water content has a decisive influence on dielectric properties and those might be assessed by the study of the wave properties that are derived by using GPR. Therefore, the waterfront advance will result in a change on wave parameters. In line with this, this research is focused on the development of specific processing algorithms necessary to understand how the water penetrates and how the wave parameters will be affected regarding the location of the antenna in reference to the water absorption direction. For this purpose, concrete samples were manufactured, which after curing (90 days) and oven

  15. Mitochondrial Ca2+-induced Ca2+ Release Mediated by the Ca2+ Uniporter

    PubMed Central

    Montero, Mayte; Alonso, Maria Teresa; Albillos, Almudena; García-Sancho, Javier; Alvarez, Javier

    2001-01-01

    We have reported that a population of chromaffin cell mitochondria takes up large amounts of Ca2+ during cell stimulation. The present study focuses on the pathways for mitochondrial Ca2+ efflux. Treatment with protonophores before cell stimulation abolished mitochondrial Ca2+ uptake and increased the cytosolic [Ca2+] ([Ca2+]c) peak induced by the stimulus. Instead, when protonophores were added after cell stimulation, they did not modify [Ca2+]c kinetics and inhibited Ca2+ release from Ca2+-loaded mitochondria. This effect was due to inhibition of mitochondrial Na+/Ca2+ exchange, because blocking this system with CGP37157 produced no further effect. Increasing extramitochondrial [Ca2+]c triggered fast Ca2+ release from these depolarized Ca2+-loaded mitochondria, both in intact or permeabilized cells. These effects of protonophores were mimicked by valinomycin, but not by nigericin. The observed mitochondrial Ca2+-induced Ca2+ release response was insensitive to cyclosporin A and CGP37157 but fully blocked by ruthenium red, suggesting that it may be mediated by reversal of the Ca2+ uniporter. This novel kind of mitochondrial Ca2+-induced Ca2+ release might contribute to Ca2+ clearance from mitochondria that become depolarized during Ca2+ overload. PMID:11160823

  16. COST Action TU1208 "Civil Engineering Applications of Ground Penetrating Radar": first-year activities and results

    NASA Astrophysics Data System (ADS)

    Pajewski, Lara; Benedetto, Andrea; Loizos, Andreas; Slob, Evert; Tosti, Fabio

    2014-05-01

    This work aims at presenting the first-year activities and results of COST (European COoperation in Science and Technology) Action TU1208 "Civil Engineering Applications of Ground Penetrating Radar". This Action was launched in April 2013 and will last four years. The principal aim of COST Action TU1208 is to exchange and increase scientific-technical knowledge and experience of GPR techniques in civil engineering, whilst simultaneously promoting throughout Europe the effective use of this safe and non-destructive technique in the monitoring of infrastructures and structures. Moreover, the Action is oriented to the following specific objectives and expected deliverables: (i) coordinating European scientists to highlight problems, merits and limits of current GPR systems; (ii) developing innovative protocols and guidelines, which will be published in a handbook and constitute a basis for European standards, for an effective GPR application in civil- engineering tasks; safety, economic and financial criteria will be integrated within the protocols; (iii) integrating competences for the improvement and merging of electromagnetic scattering techniques and of data- processing techniques; this will lead to a novel freeware tool for the localization of buried objects, shape-reconstruction and estimation of geophysical parameters useful for civil engineering needs; (iv) networking for the design, realization and optimization of innovative GPR equipment; (v) comparing GPR with different NDT techniques, such as ultrasonic, radiographic, liquid-penetrant, magnetic-particle, acoustic-emission and eddy-current testing; (vi) comparing GPR technology and methodology used in civil engineering with those used in other fields; (vii) promotion of a more widespread, advanced and efficient use of GPR in civil engineering; and (viii) organization of a high-level modular training program for GPR European users. Four Working Groups (WGs) carry out the research activities. The first WG

  17. Viruses, Mark Thy Message Well.

    PubMed

    Ye, Fengchun; Karn, Jonathan

    2016-05-11

    Post-transcriptional m(6)A methylation of RNA has profound effects on RNA splicing, export, stability, and translation. A recent study by Lichinchi et al. (2016) and one in this issue of Cell Host & Microbe by Kennedy et al. (2016) demonstrate that HIV mRNA is extensively m(6)A methylated, which promotes efficient virus replication. PMID:27173921

  18. Routes of Ca2+ Shuttling during Ca2+ Oscillations

    PubMed Central

    Pecze, László; Blum, Walter; Schwaller, Beat

    2015-01-01

    In some cell types, Ca2+ oscillations are strictly dependent on Ca2+ influx across the plasma membrane, whereas in others, oscillations also persist in the absence of Ca2+ influx. We observed that, in primary mesothelial cells, the plasmalemmal Ca2+ influx played a pivotal role. However, when the Ca2+ transport across the plasma membrane by the “lanthanum insulation method” was blocked prior to the induction of the serum-induced Ca2+ oscillations, mitochondrial Ca2+ transport was found to be able to substitute for the plasmalemmal Ca2+ exchange function, thus rendering the oscillations independent of extracellular Ca2+. However, in a physiological situation, the Ca2+-buffering capacity of mitochondria was found not to be essential for Ca2+ oscillations. Moreover, brief spontaneous Ca2+ changes were observed in the mitochondrial Ca2+ concentration without apparent changes in the cytosolic Ca2+ concentration, indicating the presence of a mitochondrial autonomous Ca2+ signaling mechanism. In the presence of calretinin, a Ca2+-buffering protein, the amplitude of cytosolic spikes during oscillations was decreased, and the amount of Ca2+ ions taken up by mitochondria was reduced. Thus, the increased calretinin expression observed in mesothelioma cells and in certain colon cancer might be correlated to the increased resistance of these tumor cells to proapoptotic/pronecrotic signals. We identified and characterized (experimentally and by modeling) three Ca2+ shuttling pathways in primary mesothelial cells during Ca2+ oscillations: Ca2+ shuttled between (i) the endoplasmic reticulum (ER) and mitochondria, (ii) the ER and the extracellular space, and (iii) the ER and cytoplasmic Ca2+ buffers. PMID:26396196

  19. Two-way WKB Approximation Applied to GPR - COST Action TU1208

    NASA Astrophysics Data System (ADS)

    Prokopovich, Igor; Popov, Alexei; Marciniak, Marian; Pajewski, Lara

    2016-04-01

    account. In this work we study bistatic EM pulse probing of a horizontally layered medium in a 2D case. Coupled WKB equations set describing both forward and backward waves are derived and solved analytically. The comparison of our semi-analytical solutions with numerical calculations by gprMax software [4] demonstrates a good agreement, being hundreds of times faster than the letter. Our numerical results explain the protracted return pulses in the low-frequency GPR data. As an example, we discuss the experimental data obtained during the GPR mission in search of a big fragment of Chelyabinsk meteorite under a thick silt layer at the bottom of Chebarcul' Lake [5]. Acknowledgement The Authors are grateful to the European Cooperation in Science and Technology (www.cost.eu) facilitating this work by a Short-Term Scientific Missions (STSM) within the framework of the Action TU1208 "Civil engineering applications of Ground Penetrating Radar" (www.GPRadar.eu). References 1. H. Bremmer "Propagation of electromagnetic waves", in Handbuch der Physik, S. Flugge, Ed. Berlin-Goettingen-Heidelberg: Springer, 1958, pp. 423-639 2. L.M. Brekhovskikh, Waves in Stratified Media (in Russian). Moscow: USSR Academy of Sciences, 1957. 3. V.A.Vinogradov, V.V. Kopeikin, A.V. Popov, "An Approximate Solution of 1D Inverse Problem", in Proc. 10th Internat. Conf. on GPR, 21-24 June, 2004, Delft, The Netherlands 4. A. Giannopoulos, "Modelling ground penetrating radar by GprMax", Construction and Building Materials, vol. 19, no. 10, pp. 755-762, 2005, doi: 10.1016/j.conbuildmat.2005.06.007 5. V. V. Kopeikin , V. D. Kuznetsov, P. A. Morozov, A. V. Popov et al., "Ground penetrating radar investigation of the supposed fall site of a fragment of the Chelyabinsk meteorite in Lake Chebarkul'", Geochemistry International, vol. 51, no. 7, pp. 575-582, 2013, doi: 10.1134/S0016702913070112

  20. Formulations of the endophytic bacterium Bacillus subtilis Tu-100 suppress Sclerotinia sclerotiorum on oilseed rape and improve plant vigor in field trials conducted at separate locations

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Sclerotinia sclerotiorum causes serious yield losses in crops in The People’s Republic of China. Two formulations of oilseed rape seed containing the endophytic bacterium Bacillus subtilis Tu-100 were evaluated for suppression of this pathogen in field trials conducted at two independent locations....

  1. Xgrip109: A γ Tubulin–Associated Protein with an Essential Role in γ Tubulin Ring Complex (γTuRC) Assembly and Centrosome Function

    PubMed Central

    Martin, Ona C.; Gunawardane, Ruwanthi N.; Iwamatsu, Akihiro; Zheng, Yixian

    1998-01-01

    Previous studies indicate that γ tubulin ring complex (γTuRC) can nucleate microtubule assembly and may be important in centrosome formation. γTuRC contains approximately eight subunits, which we refer to as Xenopus gamma ring proteins (Xgrips), in addition to γ tubulin. We found that one γTuRC subunit, Xgrip109, is a highly conserved protein, with homologues present in yeast, rice, flies, zebrafish, mice, and humans. The yeast Xgrip109 homologue, Spc98, is a spindle–pole body component that interacts with γ tubulin. In vertebrates, Xgrip109 identifies two families of related proteins. Xgrip109 and Spc98 have more homology to one family than the other. We show that Xgrip109 is a centrosomal protein that directly interacts with γ tubulin. We have developed a complementation assay for centrosome formation using demembranated Xenopus sperm and Xenopus egg extract. Using this assay, we show that Xgrip109 is necessary for the reassembly of salt-disrupted γTuRC and for the recruitment of γ tubulin to the centrosome. Xgrip109, therefore, is essential for the formation of a functional centrosome. PMID:9566968

  2. :Examination of Sulfate production by CB05TU, RACM2 & RACM2 with SCI initiated SO2,oxidation in the Northern Hemisphere"

    EPA Science Inventory

    We employ the Community Multiscale Air Quality model to examine tropospheric sulfateproduction in the northern hemisphere using the Carbon Bond 2005 chemical mechanism withupdated toluene chemistry (CB05TU) and the Regional Atmospheric Chemistry Mechanism(RACM2) without and with ...

  3. Tinnitus assessment by means of standardized self-report questionnaires: Psychometric properties of the Tinnitus Questionnaire (TQ), the Tinnitus Handicap Inventory (THI), and their short versions in an international and multi-lingual sample

    PubMed Central

    2012-01-01

    Background Tinnitus research in an international context requires standardized and validated questionnaires in different languages. The aim of the present set of analyses was the reassessment of basic psychometric properties according to classical test theory of self-report instruments that are being used within the multicentre Tinnitus Research Initiative (TRI) database project. Methods 1318 patients of the TRI Database were eligible for the analyses. The basic psychometric properties reliability, validity, and sensitivity of Tinnitus Handicap Inventory (THI), Tinnitus Questionnaire (TQ) and Tinnitus Beeinträchtigungs Fragebogen (i.e., Tinnitus Impairment Questionnaire, TBF-12) were assessed by the use of Cronbach’s alpha, corrected item-total correlations, correlation coefficients and standardized response means. Results Throughout the languages, all questionnaires showed high internal consistencies (Cronbach’s alpha > 0.79) and solid item-total correlations, as well as high correlations among themselves (around 0.8) and in combination with the self-reported tinnitus severity. However, some paradoxical correlations between individual items of the TBF-12, constructed as a shortform of the THI, and the corresponding THI-items were seen. Standardized Response Means (SRM) were low if tinnitus did not change, and between 0.3 and 1.09 for improved or worsened tinnitus complaints, indicating the sensitivity of the measures. Conclusions All investigated instruments have high internal consistency, high convergence and discriminant validity and good change sensitivity in an unselected large multinational clinical sample and thus appear appropriate to evaluate the effects of tinnitus treatments in a cross-cultural context. PMID:23078754

  4. Topological organization of CA3-to-CA1 excitation.

    PubMed

    Hongo, Yoshie; Ogawa, Koichi; Takahara, Yuji; Takasu, Keiko; Royer, Sebastien; Hasegawa, Minoru; Sakaguchi, Gaku; Ikegaya, Yuji

    2015-09-01

    The CA1-projecting axons of CA3 pyramidal cells, called Schaffer collaterals, constitute one of the major information flow routes in the hippocampal formation. Recent anatomical studies have revealed the non-random structural connectivity between CA3 and CA1, but little is known regarding the functional connectivity (i.e. how CA3 network activity is functionally transmitted downstream to the CA1 network). Using functional multi-neuron calcium imaging of rat hippocampal slices, we monitored the spatiotemporal patterns of spontaneous CA3 and CA1 burst activity under pharmacological GABAergic blockade. We found that spatially clustered CA3 activity patterns were transformed into layered CA1 activity sequences. Specifically, synchronized bursts initiated from multiple hot spots in CA3 ensembles, and CA1 neurons located deeper in the pyramidal cell layer were recruited during earlier phases of the burst events. The order of these sequential activations was maintained across the bursts, but the sequence velocity varied depending on the inter-burst intervals. Thus, CA3 axons innervate CA1 neurons in a highly topographical fashion. PMID:26036915

  5. UV - RIVERSIDE CA

    EPA Science Inventory

    Brewer 112 is located in Riverside CA, measuring ultraviolet solar radiation. Irradiance and column ozone are derived from this data. Ultraviolet solar radiation is measured with a Brewer Mark IV, single-monochrometer, spectrophotometer manufactured by SCI-TEC Instruments, Inc. o...

  6. [Tuberculosis Laboratory Surveillance Network (TuLSA) study group. The first step for national tuberculosis laboratory surveillance: Ankara, 2011].

    PubMed

    Sezen, Figen; Albayrak, Nurhan; Özkara, Şeref; Karagöz, Alper; Alp, Alpaslan; Duyar Ağca, Filiz; İnan Süer, Asiye; Müderris, Tuba; Ceyhan, İsmail; Durmaz, Rıza; Ertek, Mustafa

    2015-04-01

    The most effective method for monitoring country-level drug resistance frequency and to implement the necessary control measures is the establishment of a laboratory-based surveillance system. The aim of this study was to summarize the follow up trend of the drug-resistant tuberculosis (TB) cases, determine the load of resistance and evaluate the capacities of laboratories depending on laboratory quality assurance system for the installation work of National Tuberculosis Laboratory Surveillance Network (TuLSA) which has started in Ankara in 2011. TuLSA studies was carried out under the coordination of National Tuberculosis Reference Laboratory (NRL) with the participation of TB laboratories and dispensaries. Specimens of TB patients, reported from health institutions, were followed in TB laboratories, and the epidemiological information was collected from the dispensaries. One isolate per patient with the drug susceptibility test (DST) results were sent to NRL from TB laboratories and in NRL the isolates were rechecked with the genotypical (MTBDRplus, Hain Lifescience, Germany) and phenotypical (MGIT 960, BD, USA) DST methods. Molecular epidemiological analysis were also performed by spoligotyping and MIRU/VNTR. Second-line DST was applied to the isolates resistant to rifampin. A total of 1276 patients were reported between January 1st to December 31th 2011, and 335 cases were defined as "pulmonary TB from Ankara province". The mean age of those patients was 43.4 ± 20 years, and 67.5% were male. Three hundred seventeen (94.6%) patients were identified as new cases. The average sample number obtained from pulmonary TB cases was 3.26 ± 2.88, and 229 (68.3%) of them was culture positive. DST was applied to all culture positive isolates; 90.4% (207/229) of cases were susceptible to the five drugs tested (ethambutol, isoniazid, pyrazinamide, rifampicin, streptomycin). Eight (3.5%) of the isolates were multidrug-resistant (MDR-TB), while no extensively drug

  7. 3TU.Datacentrum serves data-curation needs of geodesy, hydrology, and climate science in the Netherlands

    NASA Astrophysics Data System (ADS)

    Princic, A.; Rombouts, J. P.; de Smaele, M. M. P.

    2012-04-01

    3TU.Datacentrum (datacentrum.3tu.nl) is a portal for science- and technology research data in the Netherlands. It serves data-curation needs of scientists in different disciplines of engineering and science varying from hydrology, water management, electron microscopy, to remote sensing. This data center provides long-term archive and permanent access to research data, it enables sharing and reuse of research data, and provides tools for citation of research datasets. The versatility of disciplines and versatility of needs is a challenge of the data center. The solutions offered differ in the functionality offered to the user. For a complex collection of simple datasets consisting of ongoing hydrological measurements from several project partners, using several instruments, on several locations, over a long period of time, there was a need for central storage in order to share the data and enable easy combination of data sets. The measurements are not repeatable and are also valuable for multidisciplinary research, which justified investment in the preservation and 'publishing' of the data. For a simple collection of complex datasets consisting of measurements of the meteorological radar, obtaining high resolution temporal and spatial precipitation data including those non-detectable by standard meteorological radar like drizzle, there was a need to store the data due to volume. These data come from a single instrument and from a single organization. Sharing with other scientists was difficult, which justified investment in the preservation and 'publishing' of the data. 3TU.Datacentrum offered solutions for both cases. Standardized solution to both collections is the data model: datasets, instruments, locations and time frames are all objects on their own, with own metadata and interconnected through rdf relationships. These relations are exploited in the user interface for navigation and inclusion, e.g. of instrument and location metadata in the display of

  8. FdhTU-Modulated Formate Dehydrogenase Expression and Electron Donor Availability Enhance Recovery of Campylobacter jejuni following Host Cell Infection

    PubMed Central

    Pryjma, Mark; Apel, Dmitry; Huynh, Steven; Parker, Craig T.

    2012-01-01

    Campylobacter jejuni is a food-borne bacterial pathogen that colonizes the intestinal tract and causes severe gastroenteritis. Interaction with host epithelial cells is thought to enhance severity of disease, and the ability of C. jejuni to modulate its metabolism in different in vivo and environmental niches contributes to its success as a pathogen. A C. jejuni operon comprising two genes that we designated fdhT (CJJ81176_1492) and fdhU (CJJ81176_1493) is conserved in many bacterial species. Deletion of fdhT or fdhU in C. jejuni resulted in apparent defects in adherence and/or invasion of Caco-2 epithelial cells when assessed by CFU enumeration on standard Mueller-Hinton agar. However, fluorescence microscopy indicated that each mutant invaded cells at wild-type levels, instead suggesting roles for FdhTU in either intracellular survival or postinvasion recovery. The loss of fdhU caused reduced mRNA levels of formate dehydrogenase (FDH) genes and a severe defect in FDH activity. Cell infection phenotypes of a mutant deleted for the FdhA subunit of FDH and an ΔfdhU ΔfdhA double mutant were similar to those of a ΔfdhU mutant, which likewise suggested that FdhU and FdhA function in the same pathway. Cell infection assays followed by CFU enumeration on plates supplemented with sodium sulfite abolished the ΔfdhU and ΔfdhA mutant defects and resulted in significantly enhanced recovery of all strains, including wild type, at the invasion and intracellular survival time points. Collectively, our data indicate that FdhTU and FDH are required for optimal recovery following cell infection and suggest that C. jejuni alters its metabolic potential in the intracellular environment. PMID:22636777

  9. Average Heart Rates of Hispanic and Caucasian Adolescents during Sleep: Longitudinal Analysis from the TuCASA Cohort

    PubMed Central

    Hedger-Archbold, Kristen; Sorensen, Seth T.; Goodwin, James L.; Quan, Stuart F.

    2014-01-01

    Objective: The current study describes sleeping heart rate patterns in an adolescent cohort of Hispanic and Caucasian children over approximately a 5-year period to determine how sex, ethnicity, and body mass index (BMI) contribute to sleeping heart rate patterns over time. Methods: Participants were recruited from a large urban school district in the southwest United States as part of the Tucson Children's Assessment of Sleep Apnea Study (TuCASA). Heart rate data was obtained through electrocardiogram (ECG) recordings during in-home polysomnography, approximately 5 years apart. Second-wave cohort data were analyzed to determine how age, sex, ethnicity, physical activity, and BMI contribute to average sleeping heart rates. The same variables were used to investigate how sleeping heart rate patterns change longitudinally from school-age (6–11 years) to adolescence (10–17 years) during sleep. Results: Female adolescents had significantly faster average heart rates during sleep. Sleeping heart rate decreased significantly with increasing age in the adolescent cohort. Although the Hispanic group had a statistically significant higher body mass index than Caucasians, there were no significant differences in heart rate observed between ethnicities or in those who were classified as obese (BMI ≥ 95th percentile for age). Longitudinal analysis between the school-aged and adolescent cohort revealed a significant overall decrease in heart rate across a 5-year period. Conclusions: Hispanic and Caucasian adolescents experience a similar decrease in sleeping heart rate with age. Female adolescents had significantly faster heart rates than males, and no significant differences were observed between Caucasians and Hispanics, or obese vs. nonobese adolescents. Citation: Hedger-Archbold K, Sorensen ST, Goodwin JL, Quan SF. Average heart rates of Hispanic and Caucasian adolescents during sleep: longitudinal analysis from the TuCASA cohort. J Clin Sleep Med 2014

  10. A Whitham-Theory Sonic-Boom Analysis of the TU-144 Aircraft at a Mach Number of 2.2

    NASA Technical Reports Server (NTRS)

    Mack, Robert J.

    1999-01-01

    Officially, the Tu-144 was the first supersonic-cruise, passenger-carrying aircraft to enter commercial service. Design, construction, and testing were carried out by the Soviet Union, flight certification was by the Soviet Union, and the only regular passenger flights were scheduled and flown across the territory of the Soviet Union. Although it was not introduced to international passenger service, there were many significant engineering accomplishments achieved in the design, production, and flight of this aircraft. Development of the aircraft began with a prototype stage. Systematic testing and redesign led to a production aircraft in discrete stages that measurably improved the performance of the aircraft from the starting concept to final aircraft certification. It flew in competition with the English-French Concorde for a short time, but was withdrawn from national commercial service due to a lack of interest by airlines outside the Soviet Union. NASA became interested in the Tu- 144 aircraft when it was offered for use as a flying "testbed" in the study of operating characteristics of a supersonic-cruise commercial airplane. Since it had been in supersonic-cruise service, the Tu- 144 had operational characteris'tics similar to those anticipated in the conceptual aircraft designs being studied by the United States aircraft companies. In addition to the other operational tests being conducted on the Tu-144 aircraft, it was proposed that two sets of sonic-boom pressure signature measurements be made. The first set would be made on the ground, using techniques and devices similar to those in reference I and many other subsequent studies. A second set would be made in the air with an instrumented aircraft flying close under the Tu-144 in supersonic flight. Such in-flight measurements would require pressure gages that were capable of accurately recording the flow-field overpressures generated by the Tu- 144 at relatively close distances under the vehicle

  11. Molecular characterization of the thi3 gene involved in thiamine biosynthesis in Zea mays: cDNA sequence and enzymatic and structural properties of the recombinant bifunctional protein with 4-amino-5-hydroxymethyl-2-methylpyrimidine (phosphate) kinase and thiamine monophosphate synthase activities.

    PubMed

    Rapala-Kozik, Maria; Olczak, Mariusz; Ostrowska, Katarzyna; Starosta, Agata; Kozik, Andrzej

    2007-12-01

    A thiamine biosynthesis gene, thi3, from maize Zea mays has been identified through cloning and sequencing of cDNA and heterologous overexpression of the encoded protein, THI3, in Escherichia coli. The recombinant THI3 protein was purified to homogeneity and shown to possess two essentially different enzymatic activities of HMP(-P) [4-amino-5-hydroxymethyl-2-methylpyrimidine (phosphate)] kinase and TMP (thiamine monophosphate) synthase. Both activities were characterized in terms of basic kinetic constants, with interesting findings that TMP synthase is uncompetitively inhibited by excess of one of the substrates [HMP-PP (HMP diphosphate)] and ATP. A bioinformatic analysis of the THI3 sequence suggested that these activities were located in two distinct, N-terminal kinase and C-terminal synthase, domains. Models of the overall folds of THI3 domains and the arrangements of active centre residues were obtained with the SWISS-MODEL protein modelling server, on the basis of the known three-dimensional structures of Salmonella enterica serotype Typhimurium HMP(-P) kinase and Bacillus subtilis TMP synthase. The essential roles of Gln98 and Met134 residues for HMP kinase activity and of Ser444 for TMP synthase activity were experimentally confirmed by site-directed mutagenesis. PMID:17696876

  12. A new interpretation of the deep-part of Senegal-Mauritanian Basin in the Diourbel-Thies area by integrating seismic, magnetic, gravimetric and borehole data: Implication for petroleum exploration

    NASA Astrophysics Data System (ADS)

    Ndiaye, Matar; Ngom, Papa Malick; Gorin, Georges; Villeneuve, Michel; Sartori, Mario; Medou, Joseph

    2016-09-01

    The Diourbel-Thies area is located in the centre of the onshore part of the Senegal-Mauritanian Basin (SMB). The new interpretation of old petroleum data (2-D seismic lines and drilling data of three oil wells) in the deeppart of this poorly evaluated zone, integrating gravimetric and magnetic data, has allowed the distinction of the Hercynian ante-rift phase (U1) which is distinguished from the syn-rift phase (U2) probably of Permo-Triassic to Middle Jurassic age. The syn-rift phase resulted in a series of compartments or grabens infilling aligned in a North-South direction. Tholeiitic volcanism of the Central Atlantic Magmatic Province (CAMP) is present in the syn-rift phase of the Diourbel-Thies area. The syn-rift deposits and associated volcanics allow us to surmise that the Diourbel basin represents a deeper rift basin. In comparison with other Central Atlantic Margins (CAM), the Diourbel rift basin could be one of the numerous rift basins that formed during the Permo-Triassic age. From a petroleum exploration perspective, the existence of the Diourbel rift basin is attractive because of the presence of structures that are excellent for deep gas exploration.

  13. TU-C-BRE-09: Performance Comparisons of Patient Specific IMRT QA Methodologies Using ROC Analysis

    SciTech Connect

    McKenzie, E; Balter, P; Stingo, F; Followill, D; Kry, S; Jones, J

    2014-06-15

    Purpose: To evaluate the ability of a selection of patient-specific QA methods to accurately classify IMRT plans as acceptable or unacceptable based on a multiple ion chamber (MIC) phantom. Methods: Twenty-four IMRT plans were selected (20 previously failed the institutional QA), and were measured on a MIC phantom to assess their dosimetric acceptability. These same plans were then measured using film (Kodak EDR2) and ion chamber (Wellhofer cc04), ArcCheck (Sun Nuclear), and MapCheck (Sun Nuclear) (delivered AP field-by-field, AP composite, and with original gantry angles). All gamma analyses were performed at 2%/2mm, 3%/3mm, and 5%/3mm. By using the MIC results as a gold standard, the sensitivity and specificity were calculated across a range of cut-off thresholds (% pixels passing for gamma analysis, and % dose difference for ion chamber), and were used to form ROC curves. Area under the curve (AUC) was used as a metric to quantify the performance of the various QA methods. Results: Grouping device’s AUC’s revealed two statistically significant different groups: ion chamber (AUC of 0.94), AP composite MapCheck (AUC of 0.85), ArcCheck (AUC of 0.84), and film (AUC of 0.82) were in the better performing group, while original gantry angles and AP field-by-field MapCheck (AUC of 0.65 and 0.66, respectively) matched less well with the gold standard results. Optimal cut-offs were also assessed using the ROC curves. We found that while often 90% of pixels passing is used as a criteria, the differing sensitivities of QA methods can lead to device and methodology-based optimal cutoff thresholds. Conclusion: While many methods exist to perform the same task of patient-specific IMRT QA, they utilize different strategies. This work has shown that there are inconsistencies in these methodologies in terms of their sensitivity and specificity to dosimetric acceptability. This work was supported by Public Health Service grants CA010953, CA081647, and CA21661 awarded by the

  14. COST Action TU1208 "Civil Engineering Applications of Ground Penetrating Radar": first-year activities and results

    NASA Astrophysics Data System (ADS)

    Pajewski, Lara; Benedetto, Andrea; Loizos, Andreas; Slob, Evert; Tosti, Fabio

    2014-05-01

    This work aims at presenting the first-year activities and results of COST (European COoperation in Science and Technology) Action TU1208 "Civil Engineering Applications of Ground Penetrating Radar". This Action was launched in April 2013 and will last four years. The principal aim of COST Action TU1208 is to exchange and increase scientific-technical knowledge and experience of GPR techniques in civil engineering, whilst simultaneously promoting throughout Europe the effective use of this safe and non-destructive technique in the monitoring of infrastructures and structures. Moreover, the Action is oriented to the following specific objectives and expected deliverables: (i) coordinating European scientists to highlight problems, merits and limits of current GPR systems; (ii) developing innovative protocols and guidelines, which will be published in a handbook and constitute a basis for European standards, for an effective GPR application in civil- engineering tasks; safety, economic and financial criteria will be integrated within the protocols; (iii) integrating competences for the improvement and merging of electromagnetic scattering techniques and of data- processing techniques; this will lead to a novel freeware tool for the localization of buried objects, shape-reconstruction and estimation of geophysical parameters useful for civil engineering needs; (iv) networking for the design, realization and optimization of innovative GPR equipment; (v) comparing GPR with different NDT techniques, such as ultrasonic, radiographic, liquid-penetrant, magnetic-particle, acoustic-emission and eddy-current testing; (vi) comparing GPR technology and methodology used in civil engineering with those used in other fields; (vii) promotion of a more widespread, advanced and efficient use of GPR in civil engineering; and (viii) organization of a high-level modular training program for GPR European users. Four Working Groups (WGs) carry out the research activities. The first WG

  15. Applications of GPR in archaeological prospecting and cultural heritage diagnostics: Research Perspectives in COST Action TU1208

    NASA Astrophysics Data System (ADS)

    Pajewski, Lara; Benedetto, Andrea; Schettini, Giuseppe; Soldovieri, Francesco

    2013-04-01

    Ground Penetrating Radar (GPR) is a safe, non-destructive and non-invasive imaging technique that can be effectively used for advanced inspection of composite structures and for diagnostics affecting the whole life-cycle of civil engineering works. GPR can also be successfully employed in archaeological prospecting and cultural heritage diagnostics. In many Countries, where the archeological patrimony is an outstanding value (as Egypt, Israel, Greece, Central and South America), GPR is usually employed both as a diagnostic tool for the preventive detection of archeological structures and as the most advanced instrument able to prospect geometry and shape of underground valuable sites. However many uncertainties persist, because of several difficulties and ambiguities due to the complexity of the image processing in heterogeneous environment. It is possible to identify three main areas, in GPR field, that have to be addressed in order to promote the use of this technology in archaeological prospecting and cultural heritage diagnostics. These are: a) increase of the system sensitivity to enable the usability in a wider range of conditions, archeological sites are often located in impervious and critical environments; b) research novel data processing algorithms/analysis tools for the interpretation of GPR results; c) contribute to the development of new standards and guidelines and to training of end users, that will also help to increase the awareness of operators. It is also important to further investigate and promote a combined use of GPR with other non-invasive advanced techniques, typically used in the archeological investigation. In this framework, the COST Action TU1208 "Civil Engineering Applications of Ground Penetrating Radar", proposed by a research team of "Roma Tre" University, Rome, Italy, has been approved in November 2012 and is going to start in April 2013. It is a 4-years ambitious project already involving 17 European Countries (AT, BE, CH, CZ, DE

  16. A great honor and a huge challenge for China: You-you TU getting the Nobel Prize in Physiology or Medicine

    PubMed Central

    Yuan, Da; Yang, Xue; Guo, Jun-Chao

    2016-01-01

    Public excitement over the award of the 2015 Nobel Prize in Physiology or Medicine to the Chinese medical scientist You-you TU for the discovery of a herbal anti-malarial, may mislead the Chinese people into believing that traditional Chinese herbal medicine can be used to cure all disease without any adverse effects. The aim of this paper is to explain the advantages and disadvantages of herbal traditional Chinese medicine (TCM) objectively. PMID:27143269

  17. A great honor and a huge challenge for China: You-you TU getting the Nobel Prize in Physiology or Medicine.

    PubMed

    Yuan, Da; Yang, Xue; Guo, Jun-Chao

    2016-05-01

    Public excitement over the award of the 2015 Nobel Prize in Physiology or Medicine to the Chinese medical scientist You-you TU for the discovery of a herbal anti-malarial, may mislead the Chinese people into believing that traditional Chinese herbal medi-cine can be used to cure all disease without any ad-verse effects. The aim of this paper is to explain the advantages and disadvantages of herbal traditional Chinese medicine (TCM) objectively. PMID:27143269

  18. VizieR Online Data Catalog: TU UMa light curves and maxima, CL Aur minima (Liska+, 2016)

    NASA Astrophysics Data System (ADS)

    Liska, J.; Skarka, M.; Mikulasek, Z.; Zejda, M.; Chrastina, M.

    2016-02-01

    Differential photometry for RR Lyrae star TU UMa in the 1st and 2nd file. The measurements were obtained using 24-inch and 1-inch telescopes, respectively. The observations were performed at the Masaryk University Observatory in Brno (3 nights, 24-inch), and at the private observatory in Brno (16 nights, 1-inch) in the Czech Republic from December 2013 to June 2014. Observing equipments consisted of 24-inch Newtonian telescope (600/2780mm, diameter/focal length) and a Moravian Instruments CCD camera G2-4000 with Stromgren photometric filters vby, and of 1-inch refractor (a photographic lens Sonnar 4/135mm, lens focal ratio/focal length) and ATIK 16IC CCD camera with green photometric filter with similar throughput as the Johnson V filter. Exposures were v - 60s, b - 30s, y - 30s, green - 30s. For the small aperture telescope, five frames were combined to a single image to achieve a better signal-to-noise ratio. The time resolution of a such combined frame is about 170s. CCD images were calibrated in a standard way (dark frame and flat field corrections). The C-Munipack software (Motl 2009) was used for this processing as well as for differential photometry. The comparison star BD+30 2165 was the same for both instruments, but the control stars were BD+30 2164 (for the 24-inch telescope) and HD 99593 (for the 1-inch telescope). The 3rd file contains maxima timings of TU UMa adopted from the GEOS RR Lyr database, from the latest publications, together with maxima timings determined in our study. Times of maxima were calculated from our observations, sky-surveys data (Hipparcos, NSVS, Pi of the Sky, SuperWASP), photographic measurements (project DASCH), and from several published datasets, in which the maxima were omitted or badly determined - Boenigk (1958AcA.....8...13B), Liakos, Niarchos (2011IBVS.6099....1L, 2011IBVS.5990....1L), Liu, Janes (1989ApJS...69..593L), Preston et al. (1961ApJ...133..484P). The 4th file contains minima timings of eclipsing binary CL Aur

  19. Cardiac Ca2+ signaling and Ca2+ sensitizers.

    PubMed

    Endoh, Masao

    2008-12-01

    The role of Ca2+ in cardiac excitation-contraction (E-C) coupling has been established by simultaneous measurements of contractility and Ca2+ transients by means of aequorin in intact myocardium and Ca2+ sensitive fluorescent dyes in single myocytes. The E-C coupling process can be classified into 3 processes: upstream (Ca2+ mobilization), central (Ca2+ binding to troponin C) and downstream mechanism (thin filament regulation and crossbridge cycling). These mechanisms are regulated differentially by various inotropic interventions. Positive force-frequency relationship and effects of beta-adrenoceptor stimulation, phosphodiesterase 3 inhibitors and digitalis are essentially exerted via upstream mechanism. Alpha-adrenoceptor stimulation, endothelin-1, angiotensin II, and clinically available Ca2+ sensitizers, such as levosimendan and pimobendan, act by a combination of the upstream and central/downstream mechanism. The Frank-Starling mechanism and effects of Ca2+ sensitizers such as EMD 57033 and Org 30029 are primarily induced via the central/downstream mechanism. Whereas the upstream and central mechanisms are markedly suppressed in failing myocytes and under acidotic conditions, Ca2+ sensitizers such as EMD 57033 and Org 30029 can induce cardiotonic effects under such conditions. Ca2+ sensitizers have high therapeutic potential for the treatment of contractile dysfunction in congestive heart failure and ischemic heart diseases, because they have energetic advantages and less risk of Ca2+ overload and can maintain effectiveness under pathological conditions. PMID:18981594

  20. Intracellular BK(Ca) (iBK(Ca)) channels.

    PubMed

    Singh, Harpreet; Stefani, Enrico; Toro, Ligia

    2012-12-01

    The large conductance calcium- and voltage-activated potassium channel (BK(Ca)) is widely expressed at the plasma membrane. This channel is involved in a variety of fundamental cellular functions including excitability, smooth muscle contractility, and Ca(2+) homeostasis, as well as in pathological situations like proinflammatory responses in rheumatoid arthritis, and cancer cell proliferation. Immunochemical, biochemical and pharmacological studies from over a decade have intermittently shown the presence of BK(Ca) in intracellular organelles. To date, intracellular BK(Ca) (iBK(Ca)) has been localized in the mitochondria, endoplasmic reticulum, nucleus and Golgi apparatus but its functional role remains largely unknown except for the mitochondrial BK(Ca) whose opening is thought to play a role in protecting the heart from ischaemic injury. In the nucleus, pharmacology suggests a role in regulating nuclear Ca(2+), membrane potential and eNOS expression. Establishing the molecular correlates of iBK(Ca), the mechanisms defining iBK(Ca) organelle-specific targeting, and their modulation are challenging questions. This review summarizes iBK(Ca) channels, their possible functions, and efforts to identify their molecular correlates. PMID:22930268

  1. Coachella Valley, CA

    NASA Technical Reports Server (NTRS)

    2001-01-01

    These band composites, acquired on June 4, 2000, cover a 11 by 13.5 km sub-scene in the Coachella Valley, CA. The area is shown by the yellow box on the full scene in the LOWER RIGHT corner, northwest of the Salton Sea. This is a major agricultural region of California, growing fruit and produce throughout the year. Different combinations of ASTER bands help identify the different crop types. UPPER LEFT: bands 3, 2, 1 as red, green, and blue (RGB); UPPER RIGHT: bands 4, 2, 1 as RGB; LOWER LEFT: bands 4, 3, 2 as RGB. The image is centered at 33.6 degrees north latitude, 116.1 degrees west longitude.

    The U.S. science team is located at NASA's Jet Propulsion Laboratory, Pasadena, Calif. The Terra mission is part of NASA's Science Mission Directorate.

  2. Transgenic Brassica rapa plants over-expressing eIF(iso)4E variants show broad-spectrum Turnip mosaic virus (TuMV) resistance.

    PubMed

    Kim, Jinhee; Kang, Won-Hee; Hwang, Jeena; Yang, Hee-Bum; Dosun, Kim; Oh, Chang-Sik; Kang, Byoung-Cheorl

    2014-08-01

    The protein-protein interaction between VPg (viral protein genome-linked) of potyviruses and eIF4E (eukaryotic initiation factor 4E) or eIF(iso)4E of their host plants is a critical step in determining viral virulence. In this study, we evaluated the approach of engineering broad-spectrum resistance in Chinese cabbage (Brassica rapa) to Turnip mosaic virus (TuMV), which is one of the most important potyviruses, by a systematic knowledge-based approach to interrupt the interaction between TuMV VPg and B. rapa eIF(iso)4E. The seven amino acids in the cap-binding pocket of eIF(iso)4E were selected on the basis of other previous results and comparison of protein models of cap-binding pockets, and mutated. Yeast two-hybrid assay and co-immunoprecipitation analysis demonstrated that W95L, K150L and W95L/K150E amino acid mutations of B. rapa eIF(iso)4E interrupted its interaction with TuMV VPg. All eIF(iso)4E mutants were able to complement an eIF4E-knockout yeast strain, indicating that the mutated eIF(iso)4E proteins retained their function as a translational initiation factor. To determine whether these mutations could confer resistance, eIF(iso)4E W95L, W95L/K150E and eIF(iso)4E wild-type were over-expressed in a susceptible Chinese cabbage cultivar. Evaluation of the TuMV resistance of T1 and T2 transformants demonstrated that the over-expression of the eIF(iso)4E mutant forms can confer resistance to multiple TuMV strains. These data demonstrate the utility of knowledge-based approaches for the engineering of broad-spectrum resistance in Chinese cabbage. PMID:24417952

  3. TU-F-17A-03: An Analytical Respiratory Perturbation Model for Lung Motion Prediction

    SciTech Connect

    Li, G; Yuan, A; Wei, J

    2014-06-15

    training, and therefore is potentially more resilient to breathing irregularities. On-going investigation introduces airflow into the RMP model for improvement. This research is in part supported by NIH (U54CA137788/132378). AY would like to thank MSKCC summer medical student research program supported by National Cancer Institute and hosted by Department of Medical Physics at MSKCC.

  4. CA125 in Ovarian Cancer

    PubMed Central

    Urban, Nicole

    2009-01-01

    Summary Twenty five years after its discovery, circulating CA125 antigen is recommended for clinical use in the US for ovarian cancer (OC) screening of high risk women with ovaries despite its limited sensitivity and specificity. Recent findings suggest that CA125 might also serve as a predictive marker for pre-invasive OC. Methods to quantify circulating CA125 evolved towards sensitive and reliable double determinant ELISA assays. The CA125 gene, MUC16, was cloned 20 years after the protein discovery and revealed a very complex and unusual glycoprotein structure suggesting an immunological role. Recent evidence points toward CA125 function in the induction of materno-fetal tolerance through the alteration of NK phenotype. Two receptors for CA125 have been described: mesothelin and galectin-1. The specific location and functional proprieties of CA125 make it a therapeutic target of choice; clinical trials have demonstrated that anti-CA125 injections are well tolerated and suggest a potential survival benefit. PMID:20477371

  5. Ground-penetrating radar investigation of St. Leonard's Crypt under the Wawel Cathedral (Cracow, Poland) - COST Action TU1208

    NASA Astrophysics Data System (ADS)

    Benedetto, Andrea; Pajewski, Lara; Dimitriadis, Klisthenis; Avlonitou, Pepi; Konstantakis, Yannis; Musiela, Małgorzata; Mitka, Bartosz; Lambot, Sébastien; Żakowska, Lidia

    2016-04-01

    The Wawel ensemble, including the Royal Castle, the Wawel Cathedral and other monuments, is perched on top of the Wawel hill immediately south of the Cracow Old Town, and is by far the most important collection of buildings in Poland. St. Leonard's Crypt is located under the Wawel Cathedral of St Stanislaus BM and St Wenceslaus M. It was built in the years 1090-1117 and was the western crypt of the pre-existing Romanesque Wawel Cathedral, so-called Hermanowska. Pope John Paul II said his first Mass on the altar of St. Leonard's Crypt on November 2, 1946, one day after his priestly ordination. The interior of the crypt is divided by eight columns into three naves with vaulted ceiling and ended with one apse. The tomb of Bishop Maurus, who died in 1118, is in the middle of the crypt under the floor; an inscription "+ MAVRVS EPC MCXVIII +" indicates the burial place and was made in 1938 after the completion of archaeological works which resulted in the discovery of this tomb. Moreover, the crypt hosts the tombs of six Polish kings and heroes: Michał Korybut Wiśniowiecki (King of the Polish-Lithuanian Commonwealth), Jan III Sobieski (King of the Polish-Lithuanian Commonwealth and Commander at the Battle of Vienna), Maria Kazimiera (Queen of the Polish-Lithuanian Commonwealth and consort to Jan III Sobieski), Józef Poniatowski (Prince of Poland and Marshal of France), Tadeusz Kościuszko (Polish general, revolutionary and a Brigadier General in the American Revolutionary War) and Władysław Sikorski (Prime Minister of the Polish Government in Exile and Commander-in-Chief of the Polish Armed Forces). The adjacent six crypts and corridors host the tombs of the other Polish kings, from Sigismund the Old to Augustus II the Strong, their families and several Polish heroes. In May 2015, the COST (European COoperation in Science and Technology) Action TU1208 "Civil engineering applications of Ground Penetrating Radar" organised and offered a Training School (TS) on the

  6. Exchanging knowledge and working together in COST Action TU1208: Short-Term Scientific Missions on Ground Penetrating Radar

    NASA Astrophysics Data System (ADS)

    Santos Assuncao, Sonia; De Smedt, Philippe; Giannakis, Iraklis; Matera, Loredana; Pinel, Nicolas; Dimitriadis, Klisthenis; Giannopoulos, Antonios; Sala, Jacopo; Lambot, Sébastien; Trinks, Immo; Marciniak, Marian; Pajewski, Lara

    2015-04-01

    This work aims at presenting the scientific results stemming from six Short-Term Scientific Missions (STSMs) funded by the COST (European COoperation in Science and Technology) Action TU1208 'Civil Engineering Applications of Ground Penetrating Radar' (Action Chair: Lara Pajewski, STSM Manager: Marian Marciniak). STSMs are important means to develop linkages and scientific collaborations between participating institutions involved in a COST Action. Scientists have the possibility to go to an institution abroad, in order to undertake joint research and share techniques/equipment/infrastructures that may not be available in their own institution. STSMs are particularly intended for Early Stage Researchers (ESRs), i.e., young scientists who obtained their PhD since no more than 8 years when they started to be involved in the Action. Duration of a standard STSM can be from 5 to 90 days and the research activities carried out during this short stay shall specifically contribute to the achievement of the scientific objectives of the supporting COST Action. The first STSM was carried out by Lara Pajewski, visiting Antonis Giannopoulos at The University of Edinburgh (United Kingdom). The research activities focused on the electromagnetic modelling of Ground Penetrating Radar (GPR) responses to complex targets. A set of test scenarios was defined, to be used by research groups participating to Working Group 3 of COST Action TU1208, to test and compare different electromagnetic forward- and inverse-scattering methods; these scenarios were modelled by using the well-known finite-difference time-domain simulator GprMax. New Matlab procedures for the processing and visualization of GprMax output data were developed. During the second STSM, Iraklis Giannakis visited Lara Pajewski at Roma Tre University (Italy). The study was concerned with the numerical modelling of horn antennas for GPR. An air-coupled horn antenna was implemented in GprMax and tested in a realistically

  7. Dynamic buffering of mitochondrial Ca2+ during Ca2+ uptake and Na+-induced Ca2+ release

    PubMed Central

    Blomeyer, Christoph A.; Bazil, Jason N.; Stowe, David F.; Pradhan, Ranjan K.; Dash, Ranjan K.; Camara, Amadou K. S.

    2014-01-01

    In cardiac mitochondria, matrix free Ca2+ ([Ca2+]m) is primarily regulated by Ca2+ uptake and release via the Ca2+ uniporter (CU) and Na+/Ca2+ exchanger (NCE) as well as by Ca2+ buffering. Although experimental and computational studies on the CU and NCE dynamics exist, it is not well understood how matrix Ca2+ buffering affects these dynamics under various Ca2+ uptake and release conditions, and whether this influences the stoichiometry of the NCE. To elucidate the role of matrix Ca2+ buffering on the uptake and release of Ca2+, we monitored Ca2+ dynamics in isolated mitochondria by measuring both the extra-matrix free [Ca2+] ([Ca2+]e) and [Ca2+]m. A detailed protocol was developed and freshly isolated mitochondria from guinea pig hearts were exposed to five different [CaCl2] followed by ruthenium red and six different [NaCl]. By using the fluorescent probe indo-1, [Ca2+] and [Ca2+e]m were spectrofluorometrically quantified, and the stoichiometry of the NCE was determined. In addition, we measured NADH, membrane potential, matrix volume and matrix pH to monitor Ca2+-induced changes in mitochondrial bioenergetics. Our [Ca2+]e and [Ca2+]m measurements demonstrate that Ca2+ uptake and release do not show reciprocal Ca2+ dynamics in the extra-matrix and matrix compartments. This salient finding is likely caused by a dynamic Ca2+ buffering system in the matrix compartment. The Na+ - induced Ca2+ release demonstrates an electrogenic exchange via the NCE by excluding an electroneutral exchange. Mitochondrial bioenergetics were only transiently affected by Ca2+ uptake in the presence of large amounts of CaCl2, but not by Na+- induced Ca2+ release. PMID:23225099

  8. Exchanging knowledge and working together in COST Action TU1208: Short-Term Scientific Missions on Ground Penetrating Radar

    NASA Astrophysics Data System (ADS)

    Santos Assuncao, Sonia; De Smedt, Philippe; Giannakis, Iraklis; Matera, Loredana; Pinel, Nicolas; Dimitriadis, Klisthenis; Giannopoulos, Antonios; Sala, Jacopo; Lambot, Sébastien; Trinks, Immo; Marciniak, Marian; Pajewski, Lara

    2015-04-01

    This work aims at presenting the scientific results stemming from six Short-Term Scientific Missions (STSMs) funded by the COST (European COoperation in Science and Technology) Action TU1208 'Civil Engineering Applications of Ground Penetrating Radar' (Action Chair: Lara Pajewski, STSM Manager: Marian Marciniak). STSMs are important means to develop linkages and scientific collaborations between participating institutions involved in a COST Action. Scientists have the possibility to go to an institution abroad, in order to undertake joint research and share techniques/equipment/infrastructures that may not be available in their own institution. STSMs are particularly intended for Early Stage Researchers (ESRs), i.e., young scientists who obtained their PhD since no more than 8 years when they started to be involved in the Action. Duration of a standard STSM can be from 5 to 90 days and the research activities carried out during this short stay shall specifically contribute to the achievement of the scientific objectives of the supporting COST Action. The first STSM was carried out by Lara Pajewski, visiting Antonis Giannopoulos at The University of Edinburgh (United Kingdom). The research activities focused on the electromagnetic modelling of Ground Penetrating Radar (GPR) responses to complex targets. A set of test scenarios was defined, to be used by research groups participating to Working Group 3 of COST Action TU1208, to test and compare different electromagnetic forward- and inverse-scattering methods; these scenarios were modelled by using the well-known finite-difference time-domain simulator GprMax. New Matlab procedures for the processing and visualization of GprMax output data were developed. During the second STSM, Iraklis Giannakis visited Lara Pajewski at Roma Tre University (Italy). The study was concerned with the numerical modelling of horn antennas for GPR. An air-coupled horn antenna was implemented in GprMax and tested in a realistically

  9. Ground-penetrating radar investigation of St. Leonard's Crypt under the Wawel Cathedral (Cracow, Poland) - COST Action TU1208

    NASA Astrophysics Data System (ADS)

    Benedetto, Andrea; Pajewski, Lara; Dimitriadis, Klisthenis; Avlonitou, Pepi; Konstantakis, Yannis; Musiela, Małgorzata; Mitka, Bartosz; Lambot, Sébastien; Żakowska, Lidia

    2016-04-01

    The Wawel ensemble, including the Royal Castle, the Wawel Cathedral and other monuments, is perched on top of the Wawel hill immediately south of the Cracow Old Town, and is by far the most important collection of buildings in Poland. St. Leonard's Crypt is located under the Wawel Cathedral of St Stanislaus BM and St Wenceslaus M. It was built in the years 1090-1117 and was the western crypt of the pre-existing Romanesque Wawel Cathedral, so-called Hermanowska. Pope John Paul II said his first Mass on the altar of St. Leonard's Crypt on November 2, 1946, one day after his priestly ordination. The interior of the crypt is divided by eight columns into three naves with vaulted ceiling and ended with one apse. The tomb of Bishop Maurus, who died in 1118, is in the middle of the crypt under the floor; an inscription "+ MAVRVS EPC MCXVIII +" indicates the burial place and was made in 1938 after the completion of archaeological works which resulted in the discovery of this tomb. Moreover, the crypt hosts the tombs of six Polish kings and heroes: Michał Korybut Wiśniowiecki (King of the Polish-Lithuanian Commonwealth), Jan III Sobieski (King of the Polish-Lithuanian Commonwealth and Commander at the Battle of Vienna), Maria Kazimiera (Queen of the Polish-Lithuanian Commonwealth and consort to Jan III Sobieski), Józef Poniatowski (Prince of Poland and Marshal of France), Tadeusz Kościuszko (Polish general, revolutionary and a Brigadier General in the American Revolutionary War) and Władysław Sikorski (Prime Minister of the Polish Government in Exile and Commander-in-Chief of the Polish Armed Forces). The adjacent six crypts and corridors host the tombs of the other Polish kings, from Sigismund the Old to Augustus II the Strong, their families and several Polish heroes. In May 2015, the COST (European COoperation in Science and Technology) Action TU1208 "Civil engineering applications of Ground Penetrating Radar" organised and offered a Training School (TS) on the

  10. Transcriptional Profiling of Newly Generated Dentate Granule Cells Using TU Tagging Reveals Pattern Shifts in Gene Expression during Circuit Integration1,2

    PubMed Central

    Chatzi, Christina; Shen, Rongkun; Goodman, Richard H.

    2016-01-01

    Abstract Despite representing only a small fraction of hippocampal granule cells, adult-generated newborn granule cells have been implicated in learning and memory (Aimone et al., 2011). Newborn granule cells undergo functional maturation and circuit integration over a period of weeks. However, it is difficult to assess the accompanying gene expression profiles in vivo with high spatial and temporal resolution using traditional methods. Here we used a novel method [“thiouracil (TU) tagging”] to map the profiles of nascent mRNAs in mouse immature newborn granule cells compared with mature granule cells. We targeted a nonmammalian uracil salvage enzyme, uracil phosphoribosyltransferase, to newborn neurons and mature granule cells using retroviral and lentiviral constructs, respectively. Subsequent injection of 4-TU tagged nascent RNAs for analysis by RNA sequencing. Several hundred genes were significantly enhanced in the retroviral dataset compared with the lentiviral dataset. We compared a selection of the enriched genes with steady-state levels of mRNAs using quantitative PCR. Ontology analysis revealed distinct patterns of nascent mRNA expression, with newly generated immature neurons showing enhanced expression for genes involved in synaptic function, and neural differentiation and development, as well as genes not previously associated with granule cell maturation. Surprisingly, the nascent mRNAs enriched in mature cells were related to energy homeostasis and metabolism, presumably indicative of the increased energy demands of synaptic transmission and their complex dendritic architecture. The high spatial and temporal resolution of our modified TU-tagging method provides a foundation for comparison with steady-state RNA analyses by traditional transcriptomic approaches in defining the functional roles of newborn neurons. PMID:27011954